Relative Pigment Composition and Remote Sensing Reflectance of Caribbean Shallow-Water Corals

PubMed Central

Torres-Pérez, Juan L.; Guild, Liane S.; Armstrong, Roy A.; Corredor, Jorge; Zuluaga-Montero, Anabella; Polanco, Ramón

2015-01-01

Reef corals typically contain a number of pigments, mostly due to their symbiotic relationship with photosynthetic dinoflagellates. These pigments usually vary in presence and concentration and influence the spectral characteristics of corals. We studied the variations in pigment composition among seven Caribbean shallow-water Scleractinian corals by means of High Performance Liquid Chromatography (HPLC) analysis to further resolve the discrimination of corals. We found a total of 27 different pigments among the coral species, including some alteration products of the main pigments. Additionally, pigments typically found in endolithic algae were also identified. A Principal Components Analysis and a Hierarchical Cluster Analysis showed the separation of coral species based on pigment composition. All the corals were collected under the same physical environmental conditions. This suggests that pigment in the coral’s symbionts might be more genetically-determined than influenced by prevailing physical conditions of the reef. We further investigated the use of remote sensing reflectance (Rrs) as a tool for estimating the total pigment concentration of reef corals. Depending on the coral species, the Rrs and the total symbiont pigment concentration per coral tissue area correlation showed 79.5–98.5% confidence levels demonstrating its use as a non-invasive robust technique to estimate pigment concentration in studies of coral reef biodiversity and health. PMID:26619210

Relative Pigment Composition and Remote Sensing Reflectance of Caribbean Shallow-Water Corals.

PubMed

Torres-Pérez, Juan L; Guild, Liane S; Armstrong, Roy A; Corredor, Jorge; Zuluaga-Montero, Anabella; Polanco, Ramón

2015-01-01

Reef corals typically contain a number of pigments, mostly due to their symbiotic relationship with photosynthetic dinoflagellates. These pigments usually vary in presence and concentration and influence the spectral characteristics of corals. We studied the variations in pigment composition among seven Caribbean shallow-water Scleractinian corals by means of High Performance Liquid Chromatography (HPLC) analysis to further resolve the discrimination of corals. We found a total of 27 different pigments among the coral species, including some alteration products of the main pigments. Additionally, pigments typically found in endolithic algae were also identified. A Principal Components Analysis and a Hierarchical Cluster Analysis showed the separation of coral species based on pigment composition. All the corals were collected under the same physical environmental conditions. This suggests that pigment in the coral's symbionts might be more genetically-determined than influenced by prevailing physical conditions of the reef. We further investigated the use of remote sensing reflectance (Rrs) as a tool for estimating the total pigment concentration of reef corals. Depending on the coral species, the Rrs and the total symbiont pigment concentration per coral tissue area correlation showed 79.5-98.5% confidence levels demonstrating its use as a non-invasive robust technique to estimate pigment concentration in studies of coral reef biodiversity and health.

EPR imaging and HPLC characterization of the pigment-based organic free radical in black soybean seeds.

PubMed

Nakagawa, Kouichi; Maeda, Hayato

2017-02-01

We investigated the location and distribution of paramagnetic species in dry black, brown, and yellow (normal) soybean seeds using electron paramagnetic resonance (EPR), X-band (9 GHz) EPR imaging (EPRI), and HPLC. EPR primarily detected two paramagnetic species in black soybean. These two different radical species were assigned as stable organic radical and Mn 2+  species based on the g values and hyperfine structures. The signal from the stable radical was noted at g ≈ 2.00 and was relatively strong and stable. Subsequent noninvasive two-dimensional (2D) EPRI of the radical present in black soybean revealed that the stable radical was primarily located in the pigmented region of the soybean coat, with very few radicals observed in the soybean cotyledon (interior). Pigments extracted from black soybean were analyzed using HPLC. The major compound was found to be cyanidin-3-glucoside. Multi-EPR and HPLC results indicate that the stable radical was only found within the pigmented region of the soybean coat, and it could be cyanidin-3-glucoside or an oxidative decomposition product.

Identification of the Main Regulator Responsible for Synthesis of the Typical Yellow Pigment Produced by Trichoderma reesei

PubMed Central

Derntl, Christian; Rassinger, Alice; Srebotnik, Ewald; Mach, Robert L.

2016-01-01

ABSTRACT The industrially used ascomycete Trichoderma reesei secretes a typical yellow pigment during cultivation, while other Trichoderma species do not. A comparative genomic analysis suggested that a putative secondary metabolism cluster, containing two polyketide-synthase encoding genes, is responsible for the yellow pigment synthesis. This cluster is conserved in a set of rather distantly related fungi, including Acremonium chrysogenum and Penicillium chrysogenum. In an attempt to silence the cluster in T. reesei, two genes of the cluster encoding transcription factors were individually deleted. For a complete genetic proof-of-function, the genes were reinserted into the genomes of the respective deletion strains. The deletion of the first transcription factor (termed yellow pigment regulator 1 [Ypr1]) resulted in the full abolishment of the yellow pigment formation and the expression of most genes of this cluster. A comparative high-pressure liquid chromatography (HPLC) analysis of supernatants of the ypr1 deletion and its parent strain suggested the presence of several yellow compounds in T. reesei that are all derived from the same cluster. A subsequent gas chromatography/mass spectrometry analysis strongly indicated the presence of sorbicillin in the major HPLC peak. The presence of the second transcription factor, termed yellow pigment regulator 2 (Ypr2), reduces the yellow pigment formation and the expression of most cluster genes, including the gene encoding the activator Ypr1. IMPORTANCE Trichoderma reesei is used for industry-scale production of carbohydrate-active enzymes. During growth, it secretes a typical yellow pigment. This is not favorable for industrial enzyme production because it makes the downstream process more complicated and thus increases operating costs. In this study, we demonstrate which regulators influence the synthesis of the yellow pigment. Based on these data, we also provide indication as to which genes are under the control of these regulators and are finally responsible for the biosynthesis of the yellow pigment. These genes are organized in a cluster that is also found in other industrially relevant fungi, such as the two antibiotic producers Penicillium chrysogenum and Acremonium chrysogenum. The targeted manipulation of a secondary metabolism cluster is an important option for any biotechnologically applied microorganism. PMID:27520818

The Second SeaWiFS HPLC Analysis Round-Robin Experiment (SeaHARRE-2)

NASA Technical Reports Server (NTRS)

2005-01-01

Eight international laboratories specializing in the determination of marine pigment concentrations using high performance liquid chromatography (HPLC) were intercompared using in situ samples and a variety of laboratory standards. The field samples were collected primarily from eutrophic waters, although mesotrophic waters were also sampled to create a dynamic range in chlorophyll concentration spanning approximately two orders of magnitude (0.3 25.8 mg m-3). The intercomparisons were used to establish the following: a) the uncertainties in quantitating individual pigments and higher-order variables (sums, ratios, and indices); b) an evaluation of spectrophotometric versus HPLC uncertainties in the determination of total chlorophyll a; and c) the reduction in uncertainties as a result of applying quality assurance (QA) procedures associated with extraction, separation, injection, degradation, detection, calibration, and reporting (particularly limits of detection and quantitation). In addition, the remote sensing requirements for the in situ determination of total chlorophyll a were investigated to determine whether or not the average uncertainty for this measurement is being satisfied. The culmination of the activity was a validation of the round-robin methodology plus the development of the requirements for validating an individual HPLC method. The validation process includes the measurements required to initially demonstrate a pigment is validated, and the measurements that must be made during sample analysis to confirm a method remains validated. The so-called performance-based metrics developed here describe a set of thresholds for a variety of easily-measured parameters with a corresponding set of performance categories. The aggregate set of performance parameters and categories establish a) the overall performance capability of the method, and b) whether or not the capability is consistent with the required accuracy objectives.

Extraction, stability, and separation of betalains from Opuntia joconostle cv. using response surface methodology.

PubMed

Sanchez-Gonzalez, Noe; Jaime-Fonseca, Monica R; San Martin-Martinez, Eduardo; Zepeda, L Gerardo

2013-12-11

Betalains were extracted and analyzed from Opuntia joconostle (the prickly pear known as xoconostle in Mexico). For the extraction, two solvent systems were used, methanol/water and ethanol/water. A three-variable Box-Behnken statistical design was used for extraction: solvent concentration (0-80%, v/v), temperature (5-30 °C), and treatment time (10-30 min). The extraction and stability of betalains from xoconostle were studied using response surface methodology (RSM). Techniques such as UV-vis, column chromatography, and HPLC were employed for the separation and analysis of the main pigments present in the extracts. Maximum pigment concentration (92 mg/100 g of fruit) was obtained at a temperature of 15 °C and a time of 10 min for methanol/water (20:80), whereas maximum stability of the pigment was observed at pH 5 and a temperature of 25 °C. HPLC chromatograms showed the main betalains of the xoconostle characterized were betalain, betanidin, and isobetalain.

The Third SeaWiFS HPLC Analysis Round-Robin Experiment (SeaHARRE-3)

NASA Technical Reports Server (NTRS)

Hooker, Stanford B.; VanHeukelem, Laurei; Thomas, Crystal S.; Claustre, Herve; Ras, Josephine; Schluter, Louise; Clementson, Lesley; vanderLinde, Dirk; Eker-Develi, Elif; Berthon, Jean-Francois;

Analysis of polymeric phenolics in red wines using different techniques combined with gel permeation chromatography fractionation.

PubMed

Guadalupe, Zenaida; Soldevilla, Alberto; Sáenz-Navajas, María-Pilar; Ayestarán, Belén

2006-04-21

A multiple-step analytical method was developed to improve the analysis of polymeric phenolics in red wines. With a common initial step based on the fractionation of wine phenolics by gel permeation chromatography (GPC), different analytical techniques were used: high-performance liquid chromatography-diode array detection (HPLC-DAD), HPLC-mass spectrometry (MS), capillary zone electrophoresis (CZE) and spectrophotometry. This method proved to be valid for analyzing different families of phenolic compounds, such as monomeric phenolics and their derivatives, polymeric pigments and proanthocyanidins. The analytical characteristics of fractionation by GPC were studied and the method was fully validated, yielding satisfactory statistical results. GPC fractionation substantially improved the analysis of polymeric pigments by CZE, in terms of response, repeatability and reproducibility. It also represented an improvement in the traditional vanillin assay used for proanthocyanidin (PA) quantification. Astringent proanthocyanidins were also analyzed using a simple combined method that allowed these compounds, for which only general indexes were available, to be quantified.

The effect of thermal treatment on antioxidant capacity and pigment contents in separated betalain fractions.

PubMed

Mikołajczyk-Bator, Katarzyna; Pawlak, Sylwia

2016-01-01

Increased consumption of fruits and vegetables significantly reduces the risk of cardio-vascular disease. This beneficial effect on the human organism is ascribed to the antioxidant compounds these foods contain. Unfortunately, many products, particularly vegetables, need to be subjected to thermal processing before consumption. The aim of this study was to determine the effect of such thermal treatment on the antioxidant capacity and pigment contents in separated fractions of violet pigments (betacyanins) and yellow pigments (betaxanthins and betacyanins). Fractions of violet and yellow pigments were obtained by separation of betalain pigments from fresh roots of 3 red beet cultivars using column chromatography and solid phase extraction (SPE). The betalain pigment content was determined in all samples before and after thermal treatment (90°C/30 min) by spectrophotometry, according to Nilsson's method [1970] and antioxidant capacity was assessed based on ABTS. Betalain pigments in the separated fractions were identified using HPLC-MS. After thermal treatment of betacyanin fractions a slight, but statistically significant degradation of pigments was observed, while the antioxidant capacity of these fractions did not change markedly. Losses of betacyanin content amounted to 13-15% depending on the cultivar, while losses of antioxidant capacity were approx. 7%. HPLC/MS analyses showed that before heating, betanin was the dominant pigment in the betacyanin fraction, while after heating it was additionally 15-decarboxy-betanin. Isolated fractions of yellow pigments in red beets are three times less heat-resistant than betacyanin fractions. At losses of yellow pigment contents in the course of thermal treatment reaching 47%, antioxidant capacity did not change markedly (a decrease by approx. 5%). In the yellow pigment fractions neobetanin was the dominant peak in the HPLC chromatogram, while vulgaxanthin was found in a much smaller area, whereas after heating additionally 2-decarboxy-2,3-dehydro-neobetanin was detected. Both groups of betalain pigments (betacyanins and betaxanthins) exhibit antioxidant capacity before and after heating. Violet beatacyjanins are 3 times more stable when heated than yellow betaxanthins.

Characterization of phytoplankton pigments and functional community structure in the Gulf of Mannar and the Palk Bay using HPLC-CHEMTAX analysis

NASA Astrophysics Data System (ADS)

Madhu, N. V.; Ullas, N.; Ashwini, R.; Meenu, Paul; Rehitha, T. V.; Lallu, K. R.

2014-06-01

Phytoplankton marker pigments and their functional groups were identified for the first time in the Gulf of Mannar (GoM) and the Palk Bay (PB), located in the southeast coast of India using HPLC-CHEMTAX analytical techniques. The GoM generally remained more saline, productive (in terms of chlorophyll a) and less turbid than the PB during southwest and northeast monsoon periods. The diversity and concentration of marker pigments were high in the GoM, whereas the PB was characterized by high concentration of zeaxanthin, indicating the dominance of photosynthetic prokaryotes (cyanobacteria). The CHEMTAX analysis revealed that the phytoplankton biomass (chlorophyll a) in the PB was mainly derived from cyanobacterial community. However, abundance of fucoxanthin and peridinin in the GoM indicated microphytoplankton (20-200 µm) as the dominant group. The CHEMTAX results showed that more than 50% of chlorophyll a in the GoM was contributed by microphytoplankton, in particular diatoms and dinoflagellates. The substantial increase in the photoprotective carotenoids (PPCs) and photoprotection index (PI) in the PB was indicative of its low productivity, probably caused by the warm and turbid waters.

Coral Pigments: Quantification Using HPLC and Detection by Remote Sensing

NASA Technical Reports Server (NTRS)

Cottone, Mary C.

1995-01-01

Widespread coral bleaching (loss of pigments of symbiotic dinoflagellates), and the corresponding decline in coral reef health worldwide, mandates the monitoring of coral pigmentation. Samples of the corals Porites compressa and P. lobata were collected from a healthy reef at Puako, Hawaii, and chlorophyll (chl) a, peridinin, and Beta-carotene (Beta-car) were quantified using reverse-phase high performance liquid chromatography (HPLC). Detailed procedures are presented for the extraction of the coral pigments in 90% acetone, and the separation, identification, and quantification of the major zooxanthellar pigments using spectrophotometry and a modification of the HPLC system described by Mantoura and Llewellyn (1983). Beta-apo-8-carotenal was found to be inadequate as in internal standard, due to coelution with chl b and/or chl a allomer in the sample extracts. Improvements are suggested, which may result in better resolution of the major pigments and greater accuracy in quantification. Average concentrations of peridinin, chl a, and Beta-car in corals on the reef were 5.01, 8.59, and 0.29, micro-grams/cm(exp 2), respectively. Average concentrations of peridinin and Beta-car did not differ significantly between the two coral species sampled; however, the mean chl a concentration in P. compressa specimens (7.81 ,micro-grams/cm(exp 2) was significantly lower than that in P. lobata specimens (9.96 11g/cm2). Chl a concentrations determined spectrophotometrically were significantly higher than those generated through HPLC, suggesting that spectrophotometry overestimates chl a concentrations. The average ratio of chl a-to-peridinin concentrations was 1.90, with a large (53%) coefficient of variation and a significant difference between the two species sampled. Additional data are needed before conclusions can be drawn regarding average pigment concentrations in healthy corals and the consistency of the chl a/peridinin ratio. The HPLC pigment concentration values contribute to the limited database of pigment concentrations in healthy corals, from which quantitative definitions of 'healthy' vs. 'bleached' coral may emerge. They also serve as ground-truth, corresponding to fluorescence data collected from the reef at Puako using airborne remote sensing of laser induced fluorescence. Fluorescence spectra from several overflights using the NASA AOL (airborne oceanographic lidar) system show consistent chlorphyll fluorescence peaks around 685 nm, as well as consistence peaks in the 400-600 nm range which may emanate from granules in the coral tissue. These data, along with results from previous studies of coral fluorescence, suggest that remote sensing of laser-induced fluorescence may become a rapid and efficient means of monitoring coral pigmentation and coral reef bleaching.

Community analysis of pigment patterns from 37 microalgae strains reveals new carotenoids and porphyrins characteristic of distinct strains and taxonomic groups

PubMed Central

Bérard, Jean-Baptiste; Kaas, Raymond; Pasquet, Virginie; Picot, Laurent; Cadoret, Jean-Paul

2017-01-01

Phytoplankton, with an estimated 30 000 to 1 000 000 species clustered in 12 phyla, presents a high taxonomic and ecophysiological diversity, reflected by the complex distribution of pigments among the different algal classes. High performance liquid chromatography is the gold standard method for qualitative and quantitative analysis of phytoplankton pigments in seawater and culture samples, but only a few pigments can be used as robust chemotaxonomic markers. A major challenge is thus to identify new ones, characteristic of a strain, species, class or taxon that cannot be currently identified on the basis of its pigment signature. Using an optimized extraction process coupled to a HPLC de-replication strategy, we examined the pigment composition of 37 microalgae strains, representative of the broad taxonomic diversity of marine and freshwater species (excluding cyanobacteria). For each species, the major pigments already described were unambiguously identified. We also observed the presence of several minor unidentified pigments in each chromatogram. The global analysis of pigment compositions revealed a total of 124 pigments, including 98 pigments or derivatives unidentified using the standards. Absorption spectra indicated that 35 corresponded to chlorophyll/porphyrin derivatives, 57 to carotenoids and six to derivatives having both spectral signatures. Sixty-one of these unidentified or new carotenoids and porphyrin derivatives were characteristic of particular strains or species, indicating their possible use as highly specific chemotaxonomic markers capable of identifying one strain out of the 37 selected. We developed a graphical analysis using Gephi software to give a clear representation of pigment communities among the various phytoplankton strains, and to reveal strain-characteristic and shared pigments. This made it possible to reconstruct the taxonomic evolution of microalgae classes, on the basis of the conservation, loss, and/or appearance of pigments. PMID:28231253

Single cell-based analysis of torenia petal pigments by a combination of ArF excimer laser micro sampling and nano-high performance liquid chromatography (HPLC)-mass spectrometry.

PubMed

Kajiyama, Shin'ichiro; Harada, Kazuo; Fukusaki, Eiichiro; Kobayashi, Akio

2006-12-01

The molecular constituents of the petal pigments of the Torenia plant (Torenia hybrida) were analyzed on a single-cell basis by a combination of newly developed laser-microsampling and nano-flow liquid chromatography-electro spray ionization mass spectrometry (LC-ESIMS) techniques. Our method should provide a facile method for obtaining precise metabolic profiles of each cell in a single plant tissue.

Aryl Polyenes, a Highly Abundant Class of Bacterial Natural Products, Are Functionally Related to Antioxidative Carotenoids.

PubMed

Schöner, Tim A; Gassel, Sören; Osawa, Ayako; Tobias, Nicholas J; Okuno, Yukari; Sakakibara, Yui; Shindo, Kazutoshi; Sandmann, Gerhard; Bode, Helge B

2016-02-02

Bacterial pigments of the aryl polyene type are structurally similar to the well-known carotenoids with respect to their polyene systems. Their biosynthetic gene cluster is widespread in taxonomically distant bacteria, and four classes of such pigments have been found. Here we report the structure elucidation of the aryl polyene/dialkylresorcinol hybrid pigments of Variovorax paradoxus B4 by HPLC-UV-MS, MALDI-MS and NMR. Furthermore, we show for the first time that this pigment class protects the bacterium from reactive oxygen species, similarly to what is known for carotenoids. An analysis of the distribution of biosynthetic genes for aryl polyenes and carotenoids in bacterial genomes is presented; it shows a complementary distribution of these protective pigments in bacteria. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Determination of the spatial and temporal variability of phytoplankton community structure in Daya Bay via HPLC-CHEMTAX pigment analysis

NASA Astrophysics Data System (ADS)

Wang, Longhua; Ou, Linjian; Huang, Kaixuan; Chai, Chao; Wang, Zhaohui; Wang, Xiaomin; Jiang, Tao

2017-09-01

The spatial and temporal variability of the phytoplankton community structure in Daya Bay, South China Sea, were identified by using HPLC-CHEMTAX analytical techniques. The highest chlorophyll a (Chl a) concentrations were observed during summer (with an average value of 0.84 μg/L) and lowest ones during winter (with an average value of 0.33 μg/L). CHEMTAX processing revealed the seasonal succession of phytoplankton species in Daya Bay. During winter, diatoms were the dominant phytoplankton species and contributed 41.5% to total Chl a. Based on Chl a concentration, the average ratio of dinoflagellates to total phytoplankton biomass substantially increased with increasing temperature and nitrogen to phosphorus (N/P) ratio, reaching 52.2% in spring. Nutrient limitation shifted from phosphorus to nitrogen during summer. Moreover, this period was associated with the predominance of diatoms, which accounted for 71.1% of Chl a. Prasinophytes and cryptophytes were the other two dominant groups and particularly dominated during winter. Cyanobacteria became an important group during summer and autumn. Canonical correspondence analysis suggested that chrysophytes, dinoflagellates, and cryptophytes were strongly associated with high nitrate concentration, ammonium, dissolved inorganic nitrogen (DIN), and N/P ratio, and were negatively associated with temperature and phosphate. Diatoms and cyanobacteria were strongly associated with temperature, phosphate, and salinity, and are negatively influenced by nitrate, ammonium, DIN, and N/P ratio. Microscopic observations and pigment HPLC information were in good agreement for diatoms and dinoflagellates in the bay. This study demonstrated the usefulness of pigment analysis in investigating the distribution of phytoplankton groups in a complex physical environment, such as Daya Bay.

High Performance Liquid Chromatography

NASA Astrophysics Data System (ADS)

Talcott, Stephen

High performance liquid chromatography (HPLC) has many applications in food chemistry. Food components that have been analyzed with HPLC include organic acids, vitamins, amino acids, sugars, nitrosamines, certain pesticides, metabolites, fatty acids, aflatoxins, pigments, and certain food additives. Unlike gas chromatography, it is not necessary for the compound being analyzed to be volatile. It is necessary, however, for the compounds to have some solubility in the mobile phase. It is important that the solubilized samples for injection be free from all particulate matter, so centrifugation and filtration are common procedures. Also, solid-phase extraction is used commonly in sample preparation to remove interfering compounds from the sample matrix prior to HPLC analysis.

Identification of candidate amino acids involved in the formation of blue pigments in crushed garlic cloves (Allium sativum L.).

PubMed

Cho, Jungeun; Lee, Eun Jin; Yoo, Kil Sun; Lee, Seung Koo; Patil, Bhimanagouda S

2009-01-01

The color-forming ability of amino acids with thiosulfinate in crushed garlic was investigated. We developed reaction systems for generating pure blue pigments using extracted thiosulfinate from crushed garlic and onion and all 22 amino acids. Each amino acid was reacted with thiosulfinate solution and was then incubated at 60 degrees C for 3 h to generate pigments. Unknown blue pigments, responsible for discoloration in crushed garlic cloves (Allium sativum L.), were separated and tentatively characterized using high-performance liquid chromatography (HPLC) and a diode array detector ranging between 200 and 700 nm. Blue pigment solutions exhibited 2 maximal absorbance peaks at 440 nm and 580 nm, corresponding to yellow and blue, respectively, with different retention times. Our findings indicated that green discoloration is created by the combination of yellow and blue pigments. Eight naturally occurring blue pigments were separated from discolored garlic extracts using HPLC at 580 nm. This suggests that garlic discoloration is not caused by only 1 blue pigment, as reported earlier, but by as many as 8 pigments. Overall, free amino acids that formed blue pigment when reacted with thiosulfinate were glycine, arginine, lysine, serine, alanine, aspartic acid, asparagine, glutamic acid, and tyrosine. Arginine, asparagine, and glutamine had spectra that were more similar to naturally greened garlic extract.

Identification of candidate amino acids involved in the formation of pink-red pigments in onion (Allium cepa L.) juice and separation by HPLC.

PubMed

Lee, Eun Jin; Yoo, Kil Sun; Patil, Bhimanagouda S

2010-10-01

The formation of pink-red pigments ("pinking") by various amino acids was investigated by reacting amino acids with compounds present in onion juice. The unknown pink-red pigments were generated and separated using high-performance liquid chromatography (HPLC) and a diode array detector (DAD) in the range of 200 to 700 nm. To generate pink-red pigments, we developed several reaction systems using garlic alliinase, purified 1-propenyl-L-cysteine sulfoxide (1-PeCSO), onion thiosulfinate, natural onion juice, and 21 free amino acids. The compound 1-PeCSO was a key compound associated with pinking in the presence of both the alliinase and amino acids. Numerous naturally occurring pink-red pigments were detected and separated from pink onion juice using the HPLC-DAD system at 515 nm. Most free amino acids, with the exceptions of histidine, serine, and cysteine, formed various pink-red pigments when reacted with onion thiosulfinate. This observation indicated that onion pinking is caused not by a single pigment, but by many. Furthermore, more than one color compound could be produced from a single amino acid; this explains, in part, why there were many pink-red compound peaks in the chromatogram of discolored natural onion juice. We presumed that the complexity of the pink-red pigments was due to the involvement of more than 21 natural amino acids as well as several derivatives of the color products produced from each amino acid. We observed that the pinking process in onion juice is very similar to that of the greening process in crushed garlic, emphasizing that both thiosulfinate from flavor precursors and free amino acids are absolutely required for the discoloration.

Simultaneous separation and identification of oligomeric procyanidins and anthocyanin-derived pigments in raw red wine by HPLC-UV-ESI-MSn.

PubMed

Pati, S; Losito, I; Gambacorta, G; La Notte, E; Palmisano, F; Zambonin, P G

2006-07-01

Samples of raw red wine (Primitivo di Manduria, Apulia, Southern Italy) were analysed without any pre-treatment (except 1:2 dilution with water) using HPLC with detection based on UV absorbance and Electrospray Ionisation Sequential Mass Spectrometry (ESI-MSn, with n = 1-3) in a series configuration. In particular, absorbance at 520 nm was monitored for UV detection in order to identify pigments responsible for wine colour. On the other hand, two subsequent stages of MS detection based on positive ions were adopted. The first consisted of an explorative MS acquisition, aimed at the individuation of the m/z ratios for positively charged compounds; the second was based on fragmentation of the detected ions within an ion trap analyser, followed by MS/MS and, if required, MS3 acquisitions. The synergy between UV detection and MSn analysis led to the identification of 41 pigments, which can be classified into five groups: grape anthocyanins, pyranoanthocyanins, vinyl-linked anthocyanin-flavanol pigments, ethyl-bridged anthocyanin-flavanol pigments and flavanol-anthocyanin compounds. Many isomeric and oligomeric structures were found within each group. A further class of compounds, not absorbing in the visible spectrum, could be also characterised by ESI-MSn and corresponded to B-type procyanidins, i.e. proanthocyanidins arising from C4-->C8/C4-->C6 couplings between catechin or epicatechin units. In particular, oligomeric structures (from dimers to pentamers), often present with several isomers, were identified and their fragmentation patterns clarified.

Occurrence and formation kinetics of pyranomalvidin-procyanidin dimer pigment in Merlot red wine: impact of acidity and oxygen concentrations.

PubMed

Pechamat, Laurent; Zeng, Liming; Jourdes, Michael; Ghidossi, Rémy; Teissedre, Pierre-Louis

2014-02-19

Once released from red grape skins, anthocyanins undergo various chemical reactions leading to the formation of more stable pigments such as pyranoanthocyanin, as well as other derivatives. Among these pigments, pyranoanthocyanins linked directly to flavanol dimers have been detected and identified in aged Port wine but not in dry red wine. These pigments are very important with regard to the wine color evolution since they are involved in wine color evolution and stabilization. During this investigation, the occurrence in dry red wine of two pyranomalvidin-procyanidin dimer has been established by low and high resolution HPLC-UV-MS analysis. Moreover, the impact of acidity and oxygen levels on their formation in red wine has been estimated. After four months of evolution, the results showed that, for the same pH, the quantity of this pigment was correlated with oxygen concentrations. Moreover, for the same quantity of oxygen, the concentration of this pigment was related to the acidity level.

Isolation and cDNA cloning of a novel red colour-related pigment-binding protein derived from the shell of the shrimp, Litopenaeus vannamei.

PubMed

Pan, Chuang; Ishizaki, Shoichiro; Nagashima, Yuji; Gao, Jialong; Watabe, Shugo

2018-02-15

Pigment-binding proteins play important roles in crustacean shell colour change. In this study, a red colour-related pigment-binding protein, designated LvPBP75, was purified from the shell of Litopenaeus vannamei. HPLC and PAGE analysis showed that LvPBP75 was a homogeneous monomer with molecular mass of 75kDa. Peptide mass fingerprint analysis revealed that LvPBP75 belonged to hemocyanin, and the released pigment from heated LvPBP75 showed a λ max at 481nm in acetone. The significant red-colour change temperatures were detected at 30 and 80°C, respectively. Based on the determined amino acid fragments, a full-length cDNA of LvPBP75 was cloned and sequenced. The ORF encodes a protein of 662 amino acids having 80% identity with penaeidae hemocyanin. These results strongly suggest a novel function of hemocyanin, namely binding with pigment, and its involvement in L. vannamei shell colour change. Copyright © 2017 Elsevier Ltd. All rights reserved.

Characterization of [8-ethyl]-chlorophyll c3 from Emiliania huxleyi.

PubMed

Álvarez, Susana; Zapata, Manuel; Garrido, José L; Vaz, Belén

2012-06-04

We report herein the isolation and complete characterization of a member of the chlorophyll c family, designated as [8-ethyl]-chlorophyll c(3) ([8-ethyl]-chl c(3)). Structural elucidation of this pigment rested on the analysis of mono- and bidimensional NMR, UV-VIS spectroscopy and ESI-MS data, and the configuration at the 13(2) position on chiral HPLC analysis.

[Separation and identification of red pigments in natural red yolk of duck's eggs by HPLC-MS-MS].

PubMed

Liu, Liangzhong; Zhang, Min; Peng, Guanghua; Wang, Haibin; Zhang, Shenghua

2004-05-01

The natural red yolk of duck's eggs is produced by the laying duck in the lake areas in southward of China. In the laying duck breeding areas such as Honghu, Jianli, Xiantao, Tianmen and Hanchuan citys in Hubei Province, the culturists are used to feeding fresh pondweeds to the laying ducks. The yolk of duck's eggs is natural red with the chrominance reaching up to and/or above RCF (Roche Yolk Color Fan) 15. The red pigment components of natural red yolk of duck's eggs were separated and identified by thin layer chromatography (TLC), high performance liquid chromatography-mass spectrometry-mass spectrometry (HPLC-MS-MS) and high resolution electron impact-mass spectrometry (EI-MS). Four isomers of red pigments were separated by HPLC on a RP-C18 column with methanol-water (99.5:0.5, v/v) as mobile phase. The lambda(max) of the four components were 482, 488, 496, 501 nm, respectively, and all of them were single peak on chromatogram. They had the same molecular mass (Mr = 562), and had the same fragment peaks of MS2 with rhodoxanthin. The molecular formula of red pigments was determined as C40H50O2 by high resolution EI-MS. The results indicate that the red pigment is rhodoxanthin, and they are all cis-isomers of rhodoxanthin.

Identifying Phytoplankton Classes In California Reservoirs Using HPLC Pigment Analysis

NASA Astrophysics Data System (ADS)

Siddiqui, S.; Peacock, M. B.; Kudela, R. M.; Negrey, K.

2014-12-01

Few bodies of water are routinely monitored for phytoplankton composition due to monetary and time constraints, especially the less accessible bodies of water in central and southern California. These lakes and estuaries are important for economic reasons such as tourism and fishing. This project investigated the composition of phytoplankton present using pigment analysis to identify dominant phytoplankton groups. A total of 28 different sites with a wide range of salinity (0 - 60) in central and southern California were examined. These included 13 different bodies of water in central California: 6 in the Sierras, 7 in the San Francisco Bay Estuary, and 15 from southern California. The samples were analyzed using high-performance liquid-chromatography (HPLC) to quantify the pigments present (using retention time and the spectral thumbprint). Diagnostic pigments were used to indicate the phytoplankton class composition, focusing on diatoms, dinoflagellates, cryptophytes, and cyanobacteria - all key phytoplankton groups indicative of the health of the sampled reservoir. Our results indicated that cyanobacteria dominated four of the seven bodies of central California water (Mono Lake, Bridgeport Reservoir, Steamboat Slough, and Pinto Lake); cryptophytes and nannoflagellates dominated two of the central California bodies of water (Mare Island Strait and Topaz Lake); and diatoms and dinoflagellates dominated one central California body of water, Oakland Inner Harbor, comprising more than 70% of the phytoplankton present. We expect the bodies of water from Southern California to be as disparate. Though this data is only a snapshot, it has significant implications in comparing different ecosystems across California, and it has the potential to provide valuable insight into the composition of phytoplankton communities.

High Performance Liquid Chromatographic Analysis of Phytoplankton Pigments Using a C16-Amide Column

EPA Science Inventory

A reverse-phase high performance liquid chromatographic (RP-HPLC) method was developed to analyze in a single run, most polar and non-polar chlorophylls and carotenoids from marine phytoplankton. The method is based on a RP-C16-Amide column and a ternary gradient system consistin...

[Determination of azoxystrobin in tea by HPLC].

PubMed

Chonan, T

2001-08-01

A determination method has been developed for azoxystrobin in tea by HPLC. Azoxystrobin was extracted from a sample with acetone, and the extract was passed through an alumina column to remove tannin. The eluate was concentrated to ca. 25 mL and passed through a Sep-Pak Vac tC18 to remove pigments. The eluate was cleaned-up by using liquid-liquid partition, and Florisil and silica-gel columns. The HPLC analysis for azoxystrobin was carried out on a C18 column with acetonitrile-water (9:11) as the mobile phase, with ultraviolet detection at 260 nm. The recovery of azoxystrobin fortified at the level of 0.4 microgram/g was 90.2% and the limit of determination was 0.2 microgram/g.

Fast detection of leaf pigments and isoprenoids for ecophysiological studies, plant phenotyping and validating remote-sensing of vegetation.

PubMed

Junker, Laura V; Ensminger, Ingo

2016-12-01

Rapid developments in remote-sensing of vegetation and high-throughput precision plant phenotyping promise a range of real-life applications using leaf optical properties for non-destructive assessment of plant performance. Use of leaf optical properties for assessing plant performance requires the ability to use photosynthetic pigments as proxies for physiological properties and the ability to detect these pigments fast, reliably and at low cost. We describe a simple and cost-effective protocol for the rapid analysis of chlorophylls, carotenoids and tocopherols using high-performance liquid chromatography (HPLC). Many existing methods are based on the expensive solvent acetonitrile, take a long time or do not include lutein epoxide and α-carotene. We aimed to develop an HPLC method which separates all major chlorophylls and carotenoids as well as lutein epoxide, α-carotene and α-tocopherol. Using a C 30 -column and a mobile phase with a gradient of methanol, methyl-tert-butyl-ether (MTBE) and water, our method separates the above pigments and isoprenoids within 28 min. The broad applicability of our method is demonstrated using samples from various plant species and tissue types, e.g. leaves of Arabidopsis and avocado plants, several deciduous and conifer tree species, various crops, stems of parasitic dodder, fruit of tomato, roots of carrots and Chlorella algae. In comparison to previous methods, our method is very affordable, fast and versatile and can be used to analyze all major photosynthetic pigments that contribute to changes in leaf optical properties and which are of interest in most ecophysiological studies. © 2016 Scandinavian Plant Physiology Society.

Encapsulating betalains from Opuntia ficus-indica fruits by ionic gelation: Pigment chemical stability during storage of beads.

PubMed

Otálora, María Carolina; Carriazo, José Gregorio; Iturriaga, Laura; Osorio, Coralia; Nazareno, Mónica Azucena

2016-07-01

Betalain encapsulation was performed by ionic gelation as a stabilization strategy for these natural pigments. Betalains were extracted from purple cactus fruits and encapsulated in calcium-alginate and in combination of calcium alginate and bovine serum albumin. Beads were characterised by scanning electron microscopy and thermal analysis using differential scanning calorimetry and thermogravimetry. Moisture sorption isotherms were determined. Bead morphology was affected by matrix composition. Pigments storage stability was evaluated at different equilibrium relative humidity and temperatures. Pigment composition of beads was determined by HPLC-MS-MS and degradation products were also analysed after storage; betalamic acid being the major one. Both types of matrices protected the encapsulated pigments, being their storage stability better at low relative humidity than that of the non-encapsulated control material. Antiradical activities of beads were proportional to remaining betalain contents. At high relative humidity, there was no protection and low storage stability was observed in the samples. Copyright © 2016 Elsevier Ltd. All rights reserved.

Spectrophotometric Analysis of Pigments: A Critical Assessment of a High-Throughput Method for Analysis of Algal Pigment Mixtures by Spectral Deconvolution

PubMed Central

Thrane, Jan-Erik; Kyle, Marcia; Striebel, Maren; Haande, Sigrid; Grung, Merete; Rohrlack, Thomas; Andersen, Tom

2015-01-01

The Gauss-peak spectra (GPS) method represents individual pigment spectra as weighted sums of Gaussian functions, and uses these to model absorbance spectra of phytoplankton pigment mixtures. We here present several improvements for this type of methodology, including adaptation to plate reader technology and efficient model fitting by open source software. We use a one-step modeling of both pigment absorption and background attenuation with non-negative least squares, following a one-time instrument-specific calibration. The fitted background is shown to be higher than a solvent blank, with features reflecting contributions from both scatter and non-pigment absorption. We assessed pigment aliasing due to absorption spectra similarity by Monte Carlo simulation, and used this information to select a robust set of identifiable pigments that are also expected to be common in natural samples. To test the method’s performance, we analyzed absorbance spectra of pigment extracts from sediment cores, 75 natural lake samples, and four phytoplankton cultures, and compared the estimated pigment concentrations with concentrations obtained using high performance liquid chromatography (HPLC). The deviance between observed and fitted spectra was generally very low, indicating that measured spectra could successfully be reconstructed as weighted sums of pigment and background components. Concentrations of total chlorophylls and total carotenoids could accurately be estimated for both sediment and lake samples, but individual pigment concentrations (especially carotenoids) proved difficult to resolve due to similarity between their absorbance spectra. In general, our modified-GPS method provides an improvement of the GPS method that is a fast, inexpensive, and high-throughput alternative for screening of pigment composition in samples of phytoplankton material. PMID:26359659

Identification and quantification of anthocyanin pigments in colored rice.

PubMed

Kim, Min-Kyoung; Kim, Han-Ah; Koh, Kwangoh; Kim, Hee-Seon; Lee, Young Sang; Kim, Yong Ho

2008-01-01

Anthocyanin pigments from varieties of black, red and wild rice were identified and quantified to evaluate their potential as nutritional function, natural colorants or functional food ingredients. Anthocyanin extraction was conducted with acidified methanol with 0.1M HCl (85:15, v/v) and identification of anthocyanin, aglycone and sugar moieties was conducted by comparison with purified standards by HPLC, Ultraviolet-Visible absorption spectrophotometer and paper chromatography. Black and wild rice showed three different types of pigments by HPLC whereas red rice variety did not show any anthocyanins. Out of three pigments detected, one (peak 2) was characterized as cyanidin-3-glucoside (C3G) by comparison of spectroscopic and chromatographic properties with an authentic standard, and another (peak 3) was tentatively identified as cyanidin-fructoside on the basis of spectroscopic properties with lambda(max) of aglycone in 1% HCl methanol at 537 nm, electrospray ionization mass spectra with major ions at 449 and 287 m/z and chromatographic properties. But another pigment (peak 1) has not been characterized. The most abundant anthocyanin in black and wild rice was C3G.

Method 447.0 - Determination of Chlorophylls a and b and Identification of Other Pigments of Interest in Marine and Freshwater Algae Using High Performance Liquid Chromatography with Visible Wavelength Detection

EPA Science Inventory

This method provides a procedure for determination of chlorophylls a (chl a) and b (chl b) found in marine and freshwater phytoplankton. Reversed phase high performance liquid chromatography (HPLC) with detection at 440 nm is used to separate the pigments from a complex pigment ...

The carotenoid pigments of a marine Bacillus firmus strain.

PubMed

Pane, L; Radin, L; Franconi, G; Carli, A

1996-01-01

As carotenoids have important biological functions, it is important to discover new natural sources of these pigments. The bacterial strains isolated from a sea water rock pool were cultivated on marine agar containing yeast extract and identified by conventional methods. The bacterial pigments were extracted with methanol and analyzed by reversed-phase HPLC with diode array detection. The major pigment of a Bacillus firmus strain was identified as astaxanthin; the results obtained suggest potential use of this bacterium in aquaculture and in pharmaceutical field.

Characterisation of a major phytoplankton bloom in the River Thames (UK) using flow cytometry and high performance liquid chromatography.

PubMed

Moorhouse, H L; Read, D S; McGowan, S; Wagner, M; Roberts, C; Armstrong, L K; Nicholls, D J E; Wickham, H D; Hutchins, M G; Bowes, M J

2018-05-15

Recent river studies have observed rapid phytoplankton dynamics, driven by diurnal cycling and short-term responses to storm events, highlighting the need to adopt new high-frequency characterisation methods to understand these complex ecological systems. This study utilised two such analytical methods; pigment analysis by high performance liquid chromatography (HPLC) and cell counting by flow cytometry (FCM), alongside traditional chlorophyll spectrophotometry and light microscopy screening, to characterise the major phytoplankton bloom of 2015 in the River Thames, UK. All analytical techniques observed a rapid increase in chlorophyll a concentration and cell abundances from March to early June, caused primarily by a diatom bloom. Light microscopy identified a shift from pennate to centric diatoms during this period. The initial diatom bloom coincided with increased HPLC peridinin concentrations, indicating the presence of dinoflagellates which were likely to be consuming the diatom population. The diatom bloom declined rapidly in early June, coinciding with a storm event. There were low chlorophyll a concentrations (by both HPLC and spectrophotometric methods) throughout July and August, implying low biomass and phytoplankton activity. However, FCM revealed high abundances of pico-chlorophytes and cyanobacteria through July and August, showing that phytoplankton communities remain active and abundant throughout the summer period. In combination, these techniques are able to simultaneously characterise a wider range of phytoplankton groups, with greater certainty, and provide improved understanding of phytoplankton functioning (e.g. production of UV inhibiting pigments by cyanobacteria in response to high light levels) and ecological status (through examination of pigment degradation products). Combined HPLC and FCM analyses offer rapid and cost-effective characterisation of phytoplankton communities at appropriate timescales. This will allow a more-targeted use of light microscopy to capture phytoplankton peaks or to investigate periods of rapid community succession. This will lead to greater system understanding of phytoplankton succession in response to biogeochemical drivers. Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.

Microencapsulation by spray-drying of anthocyanin pigments from Corozo ( Bactris guineensis) fruit.

PubMed

Osorio, Coralia; Acevedo, Baudilio; Hillebrand, Silke; Carriazo, José; Winterhalter, Peter; Morales, Alicia Lucía

2010-06-09

The anthocyanins of Bactris guineensis fruit were isolated with the aid of high-speed countercurrent chromatography (HSCCC) and preparative HPLC, and their chemical structures were elucidated by using spectroscopic methods. Among the identified pigments, cyanidin-3-rutinoside and cyanidin-3-glucoside were characterized as major constituents (87.9%). Peonidin-3-rutinoside, peonidin-3-glucoside, cyanidin-3-(6-O-malonyl)glucoside, and cyanidin-3-sambubioside were present in minor amounts. Four anthocyanin ethanolic extracts (AEEs) were obtained by osmotic dehydration and Soxhlet extraction and physicochemically characterized. The composition of anthocyanins was monitored by HPLC-PDA. The extracts with the highest anthocyanin content were subjected to the spray-drying process with maltodextrin. The so-obtained spray-dried powders were analyzed by scanning electron microscopy (SEM) and found to consist of spherical particles <50 microm in size. The anthocyanin composition was similar to that of the fruit. The microencapsulated powders were analyzed by thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC), revealing that they are quite stable until 100 degrees C. Storage stability tests of microcapsules showed that the release of anthocyanin pigments follows pseudo-first-order kinetics and that the process rate is increased by temperature and humidity. The most suitable conditions for storage were below 37 degrees C and <76% relative humidity, respectively.

Phytoplankton-Specific Response to Enrichment of Phosphorus-Rich Surface Waters with Ammonium, Nitrate, and Urea

PubMed Central

Donald, Derek B.; Bogard, Matthew J.; Finlay, Kerri; Bunting, Lynda; Leavitt, Peter R.

2013-01-01

Supply of anthropogenic nitrogen (N) to the biosphere has tripled since 1960; however, little is known of how in situ response to N fertilisation differs among phytoplankton, whether species response varies with the chemical form of N, or how interpretation of N effects is influenced by the method of analysis (microscopy, pigment biomarkers). To address these issues, we conducted two 21-day in situ mesocosm (3140 L) experiments to quantify the species- and genus-specific responses of phytoplankton to fertilisation of P-rich lake waters with ammonium (NH4 +), nitrate (NO3 −), and urea ([NH2]2CO). Phytoplankton abundance was estimated using both microscopic enumeration of cell densities and high performance liquid chromatographic (HPLC) analysis of algal pigments. We found that total algal biomass increased 200% and 350% following fertilisation with NO3 − and chemically-reduced N (NH4 +, urea), respectively, although 144 individual taxa exhibited distinctive responses to N, including compound-specific stimulation (Planktothrix agardhii and NH4 +), increased biomass with chemically-reduced N alone (Scenedesmus spp., Coelastrum astroideum) and no response (Aphanizomenon flos-aquae, Ceratium hirundinella). Principle components analyses (PCA) captured 53.2–69.9% of variation in experimental assemblages irrespective of the degree of taxonomic resolution of analysis. PCA of species-level data revealed that congeneric taxa exhibited common responses to fertilisation regimes (e.g., Microcystis aeruginosa, M. flos-aquae, M. botrys), whereas genera within the same division had widely divergent responses to added N (e.g., Anabaena, Planktothrix, Microcystis). Least-squares regression analysis demonstrated that changes in phytoplankton biomass determined by microscopy were correlated significantly (p<0.005) with variations in HPLC-derived concentrations of biomarker pigments (r 2 = 0.13–0.64) from all major algal groups, although HPLC tended to underestimate the relative abundance of cyanobacteria. Together, these findings show that while fertilisation of P-rich lakes with N can increase algal biomass, there is substantial variation in responses of genera and divisions to specific chemical forms of added N. PMID:23349705

Phytoplankton-specific response to enrichment of phosphorus-rich surface waters with ammonium, nitrate, and urea.

PubMed

Donald, Derek B; Bogard, Matthew J; Finlay, Kerri; Bunting, Lynda; Leavitt, Peter R

2013-01-01

Supply of anthropogenic nitrogen (N) to the biosphere has tripled since 1960; however, little is known of how in situ response to N fertilisation differs among phytoplankton, whether species response varies with the chemical form of N, or how interpretation of N effects is influenced by the method of analysis (microscopy, pigment biomarkers). To address these issues, we conducted two 21-day in situ mesocosm (3140 L) experiments to quantify the species- and genus-specific responses of phytoplankton to fertilisation of P-rich lake waters with ammonium (NH(4)(+)), nitrate (NO(3)(-)), and urea ([NH(2)](2)CO). Phytoplankton abundance was estimated using both microscopic enumeration of cell densities and high performance liquid chromatographic (HPLC) analysis of algal pigments. We found that total algal biomass increased 200% and 350% following fertilisation with NO(3)(-) and chemically-reduced N (NH(4)(+), urea), respectively, although 144 individual taxa exhibited distinctive responses to N, including compound-specific stimulation (Planktothrix agardhii and NH(4)(+)), increased biomass with chemically-reduced N alone (Scenedesmus spp., Coelastrum astroideum) and no response (Aphanizomenon flos-aquae, Ceratium hirundinella). Principle components analyses (PCA) captured 53.2-69.9% of variation in experimental assemblages irrespective of the degree of taxonomic resolution of analysis. PCA of species-level data revealed that congeneric taxa exhibited common responses to fertilisation regimes (e.g., Microcystis aeruginosa, M. flos-aquae, M. botrys), whereas genera within the same division had widely divergent responses to added N (e.g., Anabaena, Planktothrix, Microcystis). Least-squares regression analysis demonstrated that changes in phytoplankton biomass determined by microscopy were correlated significantly (p<0.005) with variations in HPLC-derived concentrations of biomarker pigments (r(2) = 0.13-0.64) from all major algal groups, although HPLC tended to underestimate the relative abundance of cyanobacteria. Together, these findings show that while fertilisation of P-rich lakes with N can increase algal biomass, there is substantial variation in responses of genera and divisions to specific chemical forms of added N.

Tigloylshikonin, a new minor Shikonin derivative, from the roots and the commercial root extract of lithospermum erythrorhizon.

PubMed

Ito, Yusai; Onobori, Kenichi; Yamazaki, Takeshi; Kawamura, Yoko

2011-01-01

Tigloylshikonin, a new shikonin derivative esterified with tiglic acid ((E)-2-methylbut-2-enoic acid), was isolated as a minor pigment from a food colorant "Shikon color," a commercial root extract from Lithospermum erythrorhizon SIEBOLD et ZUCCARINI. The structure of tigloylshikonin was elucidated using (1)H, (13)C, the difference nuclear Overhauser effect (NOE), and 2D NMR techniques. Its stereochemistry was determined by chiral-phase HPLC analysis. Tigloylshikonin was also found in the roots of L. erythrorhizon, which indicated that this new shikonin derivative is a typical component of naphthoquinone pigments in the roots of L. erythrorhizon.

Phenotypic diversity of anthocyanins in sorghum accessions with various pericarp pigments

USDA-ARS?s Scientific Manuscript database

Anthocyanins, a sub-class of flavonoids, are natural pigments known to have functional health benefits. Sorghum is a rich source of various phytochemicals including anthocyanins. This study was to identify and quantify the profiles of anthocyanins by HPLC-DAD in the selected 25 sorghum accessions wi...

The Fourth SeaWiFS HPLC Analysis Round-Robin Experiment (SeaHARRE-4)

NASA Technical Reports Server (NTRS)

Hooker, Stanford B.; Thomas, Crystal S.; van Heukelem, Laurie; Schlueter, louise; Russ, Mary E.; Ras, Josephine; Claustre, Herve; Clementson, Lesley; Canuti, Elisabetta; Berthon, Jean-Francois;

Comparative analysis of pigments in red and yellow banana fruit.

PubMed

Fu, Xiumin; Cheng, Sihua; Liao, Yinyin; Huang, Bingzhi; Du, Bing; Zeng, Wei; Jiang, Yueming; Duan, Xuewu; Yang, Ziyin

2018-01-15

Color is an important characteristic determining the fruit value. Although ripe bananas usually have yellow peels, several banana cultivars have red peels. As details of the pigments in banana fruits are unknown, we investigated these pigments contents and compositions in the peel and pulp of red cultivar 'Hongjiaowang' and yellow cultivar 'Baxijiao' by UPLC-PDA-QTOF-MS and HPLC-PDA techniques. The 'Hongjiaowang' peel color was mainly determined by the presence of anthocyanin-containing epidermal cells. Rutinoside derivatives of cyanidin, peonidin, petunidin, and malvidin were unique to the red peel, and possibly responsible for the red color. 'Hongjiaowang' contained higher total content of carotenoids than 'Baxijiao' in both pulp and peel. Lutein, α-carotene, and β-carotene were main carotenoids, which might play a more important role than flavonoids in producing the yellow banana color owing to the properties and distribution in the fruit. The information will help us understand a complete profile of pigments in banana. Copyright © 2017 Elsevier Ltd. All rights reserved.

A new class of anthocyanin-procyanidin condensation products detected in red wine by electrospray ionization multi-stage mass spectrometry analysis.

PubMed

Sun, Baoshan; Fernandes, Tiago A; Spranger, M Isabel

2010-02-01

In our previous work, we have identified, in a model wine solution containing malvidin 3-glucoside, epicatechin and acetaldehyde, a new condensation product--hydroxylethyl-malvidin-3-glucoside-ethyl-epicatechin. The objective of this work was to verify the presence of such new condensation products in red wine. For this purpose, red wine was fractionated into various fractions by column chromatography on LiChroprep RP 18 and on Toyopearl 40 (F). The phenolic composition of each fraction was verified by HPLC-DAD and direct-infusion ESI-MS(n) analysis. In addition to the well-known anthocyanins and their acetyl and coumaroyl derivatives, and several direct and indirect anthocyanin-(epi)catechin condensation products, a new class of pigmented products, namely hydroxyethyl-anthocyanin-ethyl-flavanol compounds, have been detected in red wine. The new class of pigmented products would be expected to be the major pigments responsible for the color of aged red wine. Copyright 2010 John Wiley & Sons, Ltd.

Evaluation of phytoplankton community composition in the eutrophic Masan Bay by HPLC pigment analysis.

PubMed

Kim, Jeong Bae; Hong, Sokjin; Lee, Won-Chan; Lee, Yong-Woo; Kim, Hyung Chul; Cho, Yoonsik

2015-03-01

To assess the spatiotemporal changes in phytoplankton community composition in relation to the environment of Masan Bay, a semi-enclosed bay on the southern coast of Korea, photosynthetic pigments and environmental variables were analyzed in seawater, every month between March and November 2010. The level of dissolved inorganic nutrients was highest between July and September when the freshwater influx was at its peak, whereas chlorophyll a level was highest in April and August. Phosphate concentration was low in April (average: 0.22 +/- 0.17 microM), indicating the role of phosphate as a growth-limiting factor for phytoplankton. The results of pigment analysis indicate that dinoflagellate blooms occurred under favorable conditions, where competition with diatoms occurred. Fucoxanthin- and chlorophyll b-containing phytoplankton dominated the surface layer of Masan Bay from July to September. The composition of phytoplankton community in Masan Bay changed dramatically each month according to variations in the amount and composition of nutrients introduced through surface runoff.

Isolation and characterization of mammalian eumelanins from hair and irides.

PubMed

Novellino, L; Napolitano, A; Prota, G

2000-07-26

A new enzymatic procedure was developed for isolation of eumelanin from black human hair which might provide a substantially intact pigment for structural characterization. Sequential digestion with protease, proteinase K and papaine in the presence of dithiothreitol afforded a pigment with a 6% w/w protein content. HPLC analysis of pyrrole acids resulting from alkaline H(2)O(2) degradation, carboxyl content determination, and ferricyanide titration showed that the isolated pigment is made up of 5,6-dihydroxyindole (DHI)- and 5, 6-dihydroxyindole-2-carboxylic acid (DHICA)-derived units at a 6:1 ratio, exhibiting a significant degree of oxidative degradation. For comparison, a different eumelanin isolated from black bovine irides by a similar enzymatic procedure was analyzed. Matrix-assisted laser desorption ionization (MALDI) mass spectrometry of the final pigment provided evidence for homologous series of DHICA oligomers, while chemical analysis allowed an estimate of 2:1 DHICA/DHI-derived units in the polymer, with a substantial proportion of intact o-diphenolic functions. Iris melanin proved able to promote the Fenton oxidation of deoxyribose while hair melanin was ineffective. Overall, these results provide, for the first time, unambiguous evidence for marked structural differences of mammalian eumelanins which may be directly related to the diversity of the sites of biosynthesis and storage, as well as to functional role of these pigments.

Quantitation of chlorophylls and 22 of their colored degradation products in culinary aromatic herbs by HPLC-DAD-MS and correlation with color changes during the dehydration process.

PubMed

Lafeuille, Jean-Louis; Lefèvre, Stéphane; Lebuhotel, Julie

2014-02-26

Chlorophylls and their green and olive-brown derivatives were successfully separated from culinary herb extracts by HPLC with photodiode-array and mass spectrometry detection. The method involved a ternary gradient elution and reverse-phase separation conditions capable of resolving 24 different pigments (2 chlorophylls and 22 of their derivatives) of different polarities within 28 min. The method was applied to monitor color changes in 50 samples of culinary aromatic herbs subjected to five different drying treatments. Of the 24 pigments, 14 were key to understanding the differences between the primary degradation pathways of chlorophyll a and chlorophyll b in culinary herbs during drying processes. A color degradation ladder based on the total molar percentage of all the remaining green pigments was also proposed as a tool to measure the impact of drying treatments on aromatic herb visual aspects.

A multi-methodological approach in the study of Italian PDO "Cornetto di Pontecorvo" red sweet pepper.

PubMed

Sobolev, Anatoly P; Mannina, Luisa; Capitani, Donatella; Sanzò, Gabriella; Ingallina, Cinzia; Botta, Bruno; Fornarini, Simonetta; Crestoni, Maria Elisa; Chiavarino, Barbara; Carradori, Simone; Locatelli, Marcello; Giusti, Anna Maria; Simonetti, Giovanna; Vinci, Giuliana; Preti, Raffaella; Toniolo, Chiara; Reverberi, Massimo; Scarpari, Marzia; Parroni, Alessia; Abete, Lorena; Natella, Fausta; Di Sotto, Antonella

2018-07-30

A multi-methodological approach was applied to study red sweet peppers (Capsicum annuum L.) ecotype "Cornetto di Pontecorvo" grown in a greenhouse or in open field. This approach includes morphological analysis, chemical composition determination, and biological activity evaluation of different extracts from pepper fruits. Untargeted analyses, namely NMR spectroscopy and mass spectrometry, allowed the comprehensive pepper metabolite profile of pepper pulp, peel and seeds hydroalcoholic and organic extracts to be determined, showing the presence of sugars, organic acids, amino acids and other secondary metabolites. Targeted analyses, such as HPLC-PDA, HPLC-TLC and spectrophotometric analyses allowed polyphenols, tannins, flavonoids and pigments content to be determined. Samples quality and freshness were verified by the low content of biogenic amines and mycotoxins, as determined using HPLC-FLD and HPLC-MS, respectively. Preliminary biological results demonstrated the ability of the organic extracts to inhibit α-amylase, a key enzyme in the control of glucose metabolism. Copyright © 2018 Elsevier Ltd. All rights reserved.

HPLC-DAD-MS Profiling of Polyphenols Responsible for the Yellow-Orange Color in Apple Juices of Different French Cider Apple Varieties.

PubMed

Le Deun, Erell; Van der Werf, Remmelt; Le Bail, Gildas; Le Quéré, Jean-Michel; Guyot, Sylvain

2015-09-09

The pigments responsible for the yellow-orange coloration of apple juices have remained largely unknown up to now. Four French cider apple juices were produced in conditions similar to those used in the cider-making industry. The oxidized juices, characterized using the CIE L a b parameters, displayed various colors depending on the apple variety and native phenolic composition. HPLC-DAD-MS revealed contrasting pigment profiles related to oxidized tanning and nontanning molecules. The latter were divided into two groups according to their polarity and their visible spectra. With regard to phenolic classes, flavanol monomers and hydroxycinnamic acids played an essential role in the formation of oxidation products. Interestingly, dihydrochalcones appeared to include precursors of some yellow compounds. Indeed, the yellow pigment phloretin xyloglucoside oxidation product (PXGOPj), derived from phloretin xyloglucoside, was clearly identified in apple juices as a xyloglucose analogue of the yellow pigment phloridzin oxidation product (POPj), previously characterized in a model solution by Le Guernevé et al. (Tetrahedron Lett. 2004, 45 (35), 6673-6677).

A NEW HPLC METHOD FOR SEPARATION OF PHYTOPLANKTON PIGMENTS IN NATURAL SAMPLES

EPA Science Inventory

A new high-performance liquid chromatographic (HPLC) method was developed to analyze, in a single run, most polar and non-polar chlorophylls and carotenoids from marine phytoplankton. The method is based on a reverse-phase amide C16 (RP-amide C16) column and an elution gradient o...

Anthocyanin production in callus cultures of Cleome rosea: modulation by culture conditions and characterization of pigments by means of HPLC-DAD/ESIMS.

PubMed

Simões, Claudia; Brasil Bizarri, Carlos Henrique; da Silva Cordeiro, Lívia; Carvalho de Castro, Tatiana; Machado Coutada, Leonardo César; Ribeiro da Silva, Antônio Jorge; Albarello, Norma; Mansur, Elisabeth

2009-10-01

Leaf and stem explants of Cleome rosea formed calluses when cultured on MS medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) or 4-amino-3,5,6-trichloropicolinic acid (PIC). The highest biomass accumulation was obtained in the callus cultures initiated from stem explants on medium supplemented with 0.90 microM 2,4-D. Reddish-pink regions were observed on callus surface after 6-7 months in culture and these pigments were identified as anthocyanins. Anthocyanins production was enhanced by reducing temperature and increasing light irradiation. Pigmented calluses transferred to MS1/2 with a 1:4 ratio NH(4)(+)/NO(3)(-), 70 g L(-1) sucrose and supplementation with 0.90 microM 2,4-D maintained a high biomass accumulation and showed an increase of 150% on anthocyanin production as compared with the initial culture conditions. Qualitative analysis of calluses was performed by high performance liquid chromatography coupled to diode array detector and electrospray ionization mass spectrometry (HPLC-DAD/ESIMS). Eleven anthocyanins were characterized and the majority of them were identified as acylated cyanidins, although two peonidins were also detected. The major peak was composed by two anthocyanins, whose proposed identity were cyanidin 3-(p-coumaroyl) diglucoside-5-glucoside and cyanidin 3-(feruloyl) diglucoside-5-glucoside.

Column chromatography as a useful step in purification of diatom pigments.

PubMed

Tokarek, Wiktor; Listwan, Stanisław; Pagacz, Joanna; Leśniak, Piotr; Latowski, Dariusz

2016-01-01

Fucoxanthin, diadinoxanthin and diatoxanthin are carotenoids found in brown algae and most other heterokonts. These pigments are involved in photosynthetic and photoprotective reactions, and they have many potential health benefits. They can be extracted from diatom Phaeodactylum tricornutum by sonication, extraction with chloroform : methanol and preparative thin layer chromatography. We assessed the utility of an additional column chromatography step in purification of these pigments. This novel addition to the isolation protocol increased the purity of fucoxanthin and allowed for concentration of diadinoxanthin and diatoxanthin before HPLC separation. The enhanced protocol is useful for obtaining high purity pigments for biochemical studies.

Algal Accessory Pigment Detection Using AVIRIS Image-Derived Spectral Radiance Data

NASA Technical Reports Server (NTRS)

Richardson, Laurie L.; Ambrosia, Vincent G.

1996-01-01

Visual and derivative analyses of AVIRIS spectral data can be used to detect algal accessory pigments in aquatic communities. This capability extends the use of remote sensing for the study of aquatic ecosystems by allowing detection of taxonomically significant pigment signatures which yield information about the type of algae present. Such information allows remote sensing-based assessment of aquatic ecosystem health, as in the detection of nuisance blooms of cyanobacteria or toxic blooms of dinoflagellates. Remote sensing of aquatic systems has traditionally focused on quantification of chlorophyll a, a photoreactive (and light-harvesting) pigment which is common to all algae as well as cyanobacteria (bluegreen algae). Due to the ubiquitousness of this pigment within algae, chl a is routinely measured to estimate algal biomass both during ground-truthing and using various airborne or satellite based sensors, including AVIRIS. Within the remote sensing and aquatic sciences communities, ongoing research has been performed to detect algal accessory pigments for assessment of algal population composition. This research is based on the fact that many algal accessory pigments are taxonomically significant, and all are spectrally unique. Aquatic scientists have been refining pigment analysis techniques, primarily high performance liquid chromatography, or HPLC, to detect specific pigments as a time-saving alternative to individual algal cell identifications and counts. Remote sensing scientists are investigating the use of pigment signatures to construct pigment libraries analogous to mineral spectral libraries used in geological remote sensing applications. The accessory pigment approach has been used successfully in remote sensing using data from the Thematic Mapper, low-altitude, multiple channel scanners, field spectroradiometers and the AVIRIS hyperspectral scanner. Due to spectral and spatial resolution capabilities, AVIRIS is the sensor of choice for such studies. We present here our results on detection of algal accessory pigments using AVIRIS data.

The First SeaWiFS HPLC Analysis Round-Robin Experiment (SeaHARRE-1)

NASA Technical Reports Server (NTRS)

Hooker, Stanford B. (Editor); Firestone, Elaine R. (Editor); Claustre, Herve; Ras, Josephine; VanHeukelem, Laurie; Berthon, Jean-Francois; Targa, Cristina; vanderLinde, Dirk; Barlow, Ray; Sessions, Heather

2001-01-01

Four laboratories, which had contributed to various aspects of SeaWiFS calibration and validation activities, participated in the first SeaWiFS HPLC Analysis Round-Robin Experiment (SeaHARRE-1): Horn Point Laboratory (USA), the Joint Research Centre (Italy), the Laboratoire de Physique et Chimie Marines (France), and the Marine and Coastal Management group (South Africa). The analyses of the data are presented in Chapter 1 and the individual methods of the four groups are presented in Chapters 2-5. The average (or overall) conclusions of the round-robin are derived from 12 in situ stations occupied during a cruise in the Mediterranean Sea, although, only 11 stations are used in the analyses. The data set is composed of 12 replicates taken during each sampling opportunity with 3 replicates going to each of the 4 laboratories. The average (or overall) results from the intercomparison of 15 pigments or pigment associations are as follows (in some cases, data subsets that exclude pigments which were not analyzed by all the laboratories, or that had unusually large variances, are used to exclude a variety of problematic pigments): a) the accuracy of the four methods in determining the concentration of total chlorophyll a is 7.9%, (one method did not separate mono- and divinyl chlorophyll a, and if the samples containing significant divinyl chlorophyll a concentrations are ignored, the four methods have an accuracy of 6.7%); b) the accuracy in determining the full set of pigments is 19.1%; c) there is a reduction in accuracy of approximately - 12.2% for every decade (factor of 10) decrease in concentration (based on a data subset); d) the precision of the four methods using a subset data is 8.617( 6.2% for an edited subset); e) the repeatability of the four methods using the subset data is 9.2% (7.2%; for an edited subset, and f) the reproducibility of the four methods using the subset data is 21.31% (15.0% for an edited subset).

A Zn-porphyrin complex contributes to bright red color in Parma ham.

PubMed

Wakamatsu, J; Nishimura, T; Hattori, A

2004-05-01

The Italian traditional dry-cured ham (Parma ham) shows a stable bright red color that is achieved without the use of nitrite and/or nitrate. In this study we examined the pigment spectroscopically, fluoroscopically and by using HPLC and ESI-HR-MASS analysis. Porphyrin derivative other than acid hematin were contained in the HCl-containing acetone extract from Parma ham. A strong fluorescence peak at 588 nm and a weak fluorescence peak at 641 nm were observed. By HPLC analysis the acetone extract of Parma ham was observed at the single peak, which eluted at the same time as Zn-protoporphyrin IX and emitted fluorescence. The results of ESI-HR-MS analysis showed both agreement with the molecular weight of Zn-protoporphyrin IX and the characteristic isotope pattern caused by Zn isotopes. These results suggest that the bright red color in Parma ham is caused by Zn-protoporphyrin IX.

Interrelationships between maternal carotenoid status and newborn infant macular pigment optical density and carotenoid status.

PubMed

Henriksen, Bradley S; Chan, Gary; Hoffman, Robert O; Sharifzadeh, Mohsen; Ermakov, Igor V; Gellermann, Werner; Bernstein, Paul S

2013-08-15

Deposition of the macular pigment carotenoids lutein and zeaxanthin in the human retina occurs early in life. In this study, we examined the interrelationships of maternal carotenoid status and newborn infant macular pigment levels and systemic carotenoid status. As a secondary measure, we also evaluated the effects of intrauterine growth restriction (IUGR) on carotenoid status in term newborn infants. We measured mother and infant skin carotenoids using resonance Raman spectroscopy (RRS), serum carotenoids by HPLC, and mother breast milk carotenoids by HPLC. We measured infant macular pigment levels using noninvasive blue light reflectometry. We enrolled 30 healthy term infants, their mothers, and 10 IUGR infants and their mothers. A subset of 16 infants was imaged for macular pigment optical density (MPOD). Infant serum zeaxanthin levels correlated with MPOD (r = 0.68, P = 0.007). Mother serum zeaxanthin levels correlated with infant MPOD (r = 0.59, P = 0.032). Infant and mother serum lutein did not correlate with MPOD. Mother-infant correlations were found for total serum carotenoids (r = 0.42, P = 0.020) and skin carotenoids (r = 0.48, P = 0.001). No difference was seen between IUGR infants and controls in total serum or skin carotenoids. Mothers of IUGR infants had lower total serum carotenoids (P = 0.019) and breast milk carotenoids than controls (P = 0.006). Our findings suggest that maternal zeaxanthin status may play a more important role than lutein status in macular pigment deposition in utero. Controlled trials are needed to determine whether maternal zeaxanthin prenatal supplementation can raise infant macular pigment levels and/or improve ocular function.

Isolation and quantification of oligomeric pyranoanthocyanin-flavanol pigments from red wines by combination of column chromatographic techniques.

PubMed

He, Jingren; Santos-Buelga, Celestino; Mateus, Nuno; de Freitas, Victor

2006-11-17

A combination of column chromatography on Toyopearl gel HW-40 (S) and polyamide resin has been developed for the preparative isolation and further determination of pyranoanthocyanins of oligomeric nature formed after reaction between anthocyanins and different flavanols in a complex wine matrix. Polyamide chromatography was found to be exceptionally useful to separate oligomeric pyanoanthocyanins from other classes of wine flavonoids and polymerized pigments into an advanced state of purity for further identification and quantification by HPLC-diode array detector coupled with electrospray ionization mass spectrometry (HPLC-DAD/ESI-MS). Fractionation on Toyopearl gel chromatography allowed the separation of pyranoanthocyanins bearing the same flavanols (catechin, epicatechin and procyanidin dimers) but with different anthocyanin moieties (either acylated or non-acylated in the glucose residue) in order to allow further isolation of individual oligomeric pigments on C18 chromatography. A quantitative procedure for analyzing the major pyranoanthocyanin-flavanol derivatives in different aged wines is proposed for the first time. Results obtained showed good reproducibility and recovery regarding sample pretreatment and quantitative method for all analyzed oligomeric pyranoanthocyanins. The combination of these two chromatographic separations is likely to be applicable to the preparative isolation of other anthocyanin-derived pigments.

Pigmentation and Spectral Absorbance Signatures in Deep-Water Corals from the Trondheimsfjord, Norway

PubMed Central

Elde, Anette C.; Pettersen, Ragnhild; Bruheim, Per; Järnegren, Johanna; Johnsen, Geir

2012-01-01

The pigmentation and corresponding in vivo and in vitro absorption characteristics in three different deep-water coral species: white and orange Lophelia pertusa, Paragorgia arborea and Primnoa resedaeformis, collected from the Trondheimsfjord are described. Pigments were isolated and characterized by High-Performance Liquid Chromatography (HPLC) analysis and High-Performance Liquid Chromatography Time-Of-Flight Mass Spectrometer (LC-TOF MS). The main carotenoids identified for all three coral species were astaxanthin and a canthaxanthin-like carotenoid. Soft tissue and skeleton of orange L. pertusa contained 2 times more astaxanthin g−1 wet weight compared to white L. pertusa. White and orange L. pertusa were characterized with in vivo absorbance peaks at 409 and 473 nm, respectively. In vivo absorbance maxima for P. arborea and P. resedaeformis was typically at 475 nm. The shapes of the absorbance spectra (400–700 nm) were species-specific, indicated by in vivo, in vitro and the corresponding difference spectra. The results may provide important chemotaxonomic information for pigment when bonded to their proteins in vivo, bio-prospecting, and for in situ identification, mapping and monitoring of corals. PMID:22822381

Pigmentation and spectral absorbance signatures in deep-water corals from the Trondheimsfjord, Norway.

PubMed

Elde, Anette C; Pettersen, Ragnhild; Bruheim, Per; Järnegren, Johanna; Johnsen, Geir

2012-06-01

The pigmentation and corresponding in vivo and in vitro absorption characteristics in three different deep-water coral species: white and orange Lophelia pertusa, Paragorgia arborea and Primnoa resedaeformis, collected from the Trondheimsfjord are described. Pigments were isolated and characterized by High-Performance Liquid Chromatography (HPLC) analysis and High-Performance Liquid Chromatography Time-Of-Flight Mass Spectrometer (LC-TOF MS). The main carotenoids identified for all three coral species were astaxanthin and a canthaxanthin-like carotenoid. Soft tissue and skeleton of orange L. pertusa contained 2 times more astaxanthin g(-1) wet weight compared to white L. pertusa. White and orange L. pertusa were characterized with in vivo absorbance peaks at 409 and 473 nm, respectively. In vivo absorbance maxima for P. arborea and P. resedaeformis was typically at 475 nm. The shapes of the absorbance spectra (400-700 nm) were species-specific, indicated by in vivo, in vitro and the corresponding difference spectra. The results may provide important chemotaxonomic information for pigment when bonded to their proteins in vivo, bio-prospecting, and for in situ identification, mapping and monitoring of corals.

Rapid Estimation of Astaxanthin and the Carotenoid-to-Chlorophyll Ratio in the Green Microalga Chromochloris zofingiensis Using Flow Cytometry.

PubMed

Chen, Junhui; Wei, Dong; Pohnert, Georg

2017-07-19

The green microalga Chromochloris zofingiensis can accumulate significant amounts of valuable carotenoids, mainly natural astaxanthin, a product with applications in functional food, cosmetics, nutraceuticals, and with potential therapeutic value in cardiovascular and neurological diseases. To optimize the production of astaxanthin, it is essential to monitor the content of astaxanthin in algal cells during cultivation. The widely used HPLC (high-performance liquid chromatography) method for quantitative astaxanthin determination is time-consuming and laborious. In the present work, we present a method using flow cytometry (FCM) for in vivo determination of the astaxanthin content and the carotenoid-to-chlorophyll ratio (Car/Chl) in mixotrophic C. zofingiensis . The method is based on the assessment of fluorescent characteristics of cellular pigments. The mean fluorescence intensity (MFI) of living cells was determined by FCM to monitor pigment formation based on the correlation between MFI detected in particular channels (FL1: 533 ± 15 nm; FL2: 585 ± 20 nm; FL3: >670 nm) and pigment content in algal cells. Through correlation and regression analysis, a linear relationship was observed between MFI in FL2 (band-pass filter, emission at 585 nm in FCM) and astaxanthin content (in HPLC) and applied for predicting astaxanthin content. With similar procedures, the relationships between MFI in different channels and Car/Chl ratio in mixotrophic C. zofingiensis were also determined. Car/Chl ratios could be estimated by the ratios of MFI (FL1/FL3, FL2/FL3). FCM is thus a highly efficient and feasible method for rapid estimation of astaxanthin content in the green microalga C. zofingiensis . The rapid FCM method is complementary to the current HPLC method, especially for rapid evaluation and prediction of astaxanthin formation as it is required during the high-throughput culture in the laboratory and mass cultivation in industry.

Rapid Estimation of Astaxanthin and the Carotenoid-to-Chlorophyll Ratio in the Green Microalga Chromochloris zofingiensis Using Flow Cytometry

PubMed Central

Chen, Junhui; Pohnert, Georg

2017-01-01

The green microalga Chromochloris zofingiensis can accumulate significant amounts of valuable carotenoids, mainly natural astaxanthin, a product with applications in functional food, cosmetics, nutraceuticals, and with potential therapeutic value in cardiovascular and neurological diseases. To optimize the production of astaxanthin, it is essential to monitor the content of astaxanthin in algal cells during cultivation. The widely used HPLC (high-performance liquid chromatography) method for quantitative astaxanthin determination is time-consuming and laborious. In the present work, we present a method using flow cytometry (FCM) for in vivo determination of the astaxanthin content and the carotenoid-to-chlorophyll ratio (Car/Chl) in mixotrophic C. zofingiensis. The method is based on the assessment of fluorescent characteristics of cellular pigments. The mean fluorescence intensity (MFI) of living cells was determined by FCM to monitor pigment formation based on the correlation between MFI detected in particular channels (FL1: 533 ± 15 nm; FL2: 585 ± 20 nm; FL3: >670 nm) and pigment content in algal cells. Through correlation and regression analysis, a linear relationship was observed between MFI in FL2 (band-pass filter, emission at 585 nm in FCM) and astaxanthin content (in HPLC) and applied for predicting astaxanthin content. With similar procedures, the relationships between MFI in different channels and Car/Chl ratio in mixotrophic C. zofingiensis were also determined. Car/Chl ratios could be estimated by the ratios of MFI (FL1/FL3, FL2/FL3). FCM is thus a highly efficient and feasible method for rapid estimation of astaxanthin content in the green microalga C. zofingiensis. The rapid FCM method is complementary to the current HPLC method, especially for rapid evaluation and prediction of astaxanthin formation as it is required during the high-throughput culture in the laboratory and mass cultivation in industry. PMID:28753934

Separation of polar betalain pigments from cacti fruits of Hylocereus polyrhizus by ion-pair high-speed countercurrent chromatography.

PubMed

Wybraniec, Sławomir; Stalica, Paweł; Jerz, Gerold; Klose, Bettina; Gebers, Nadine; Winterhalter, Peter; Spórna, Aneta; Szaleniec, Maciej; Mizrahi, Yosef

2009-10-09

Polar betacyanin pigments together with betaxanthins from ripe cactus fruits of Hylocereus polyrhizus (Cactaceae) were fractionated by means of preparative ion-pair high-speed countercurrent chromatography (IP-HSCCC) also using the elution-extrusion (EE) approach for a complete pigment recovery. HSCCC separations were operated in the classical 'head-to-tail' mode with an aqueous mobile phase. Different CCC solvent systems were evaluated in respect of influence and effectiveness of fractionation capabilities to separate the occurring pigment profile of H. polyrhizus. For that reason, the additions of two different volatile ion-pair forming perfluorinated carboxylic acids (PFCA) were investigated. For a direct comparison, five samples of Hylocereus pigment extract were run on preparative scale (900 mg) in 1-butanol-acetonitrile-aqueous TFA 0.7% (5:1:6, v/v/v) and the modified systems tert.-butyl methyl ether-1-butanol-acetonitrile-aqueous PFCA (2:2:1:5, v/v/v/v) using 0.7% and 1.0% trifluoroacetic acid (TFA) or heptafluorobutyric acid (HFBA) in the aqueous phase, respectively. The chemical affinity to the organic stationary CCC solvent phases and in consequence the retention of these highly polar betalain pigments was significantly increased by the use of the more lipophilic fluorinated ion-pair reagent HFBA instead of TFA. The HFBA additions separated more effectively the typical cacti pigments phyllocactin and hylocerenin from betanin as well as their iso-forms. Unfortunately, similar K(D) ratios and selectivity factors alpha around 1.0-1.1 in all tested solvent systems proved that the corresponding diastereomers, 15S-type pigments cannot be resolved from the 15R-epimers (iso-forms). Surprisingly, additions of the stronger ion-pair reagent (HFBA) resulted in a partial separation of hylocerenin from phyllocactin which were not resolved in the other solvent systems. The pigments were detected by means of HPLC-DAD and HPLC-electrospray ionization-MS using also authentic reference materials.

Identification of Shell Colour Pigments in Marine Snails Clanculus pharaonius and C. margaritarius (Trochoidea; Gastropoda)

PubMed Central

Ito, S.; Wakamatsu, K.; Goral, T.; Edwards, N. P.; Wogelius, R. A.; Henkel, T.; de Oliveira, L. F. C.; Maia, L. F.; Strekopytov, S.; Speiser, D. I.; Marsden, J. T.

2016-01-01

Colour and pattern are key traits with important roles in camouflage, warning and attraction. Ideally, in order to begin to understand the evolution and ecology of colour in nature, it is important to identify and, where possible, fully characterise pigments using biochemical methods. The phylum Mollusca includes some of the most beautiful exemplars of biological pigmentation, with the vivid colours of sea shells particularly prized by collectors and scientists alike. Biochemical studies of molluscan shell colour were fairly common in the last century, but few of these studies have been confirmed using modern methods and very few shell pigments have been fully characterised. Here, we use modern chemical and multi-modal spectroscopic techniques to identify two porphyrin pigments and eumelanin in the shell of marine snails Clanculus pharaonius and C margaritarius. The same porphyrins were also identified in coloured foot tissue of both species. We use high performance liquid chromatography (HPLC) to show definitively that these porphyrins are uroporphyrin I and uroporphyrin III. Evidence from confocal microscopy analyses shows that the distribution of porphyrin pigments corresponds to the striking pink-red of C. pharaonius shells, as well as pink-red dots and lines on the early whorls of C. margaritarius and yellow-brown colour of later whorls. Additional HPLC results suggest that eumelanin is likely responsible for black spots. We refer to the two differently coloured porphyrin pigments as trochopuniceus (pink-red) and trochoxouthos (yellow-brown) in order to distinguish between them. Trochopuniceus and trochoxouthos were not found in the shell of a third species of the same superfamily, Calliostoma zizyphinum, despite its superficially similar colouration, suggesting that this species has different shell pigments. These findings have important implications for the study of colour and pattern in molluscs specifically, but in other taxa more generally, since this study shows that homology of visible colour cannot be assumed without identification of pigments. PMID:27367426

Identification of Shell Colour Pigments in Marine Snails Clanculus pharaonius and C. margaritarius (Trochoidea; Gastropoda).

PubMed

Williams, S T; Ito, S; Wakamatsu, K; Goral, T; Edwards, N P; Wogelius, R A; Henkel, T; de Oliveira, L F C; Maia, L F; Strekopytov, S; Jeffries, T; Speiser, D I; Marsden, J T

2016-01-01

Colour and pattern are key traits with important roles in camouflage, warning and attraction. Ideally, in order to begin to understand the evolution and ecology of colour in nature, it is important to identify and, where possible, fully characterise pigments using biochemical methods. The phylum Mollusca includes some of the most beautiful exemplars of biological pigmentation, with the vivid colours of sea shells particularly prized by collectors and scientists alike. Biochemical studies of molluscan shell colour were fairly common in the last century, but few of these studies have been confirmed using modern methods and very few shell pigments have been fully characterised. Here, we use modern chemical and multi-modal spectroscopic techniques to identify two porphyrin pigments and eumelanin in the shell of marine snails Clanculus pharaonius and C margaritarius. The same porphyrins were also identified in coloured foot tissue of both species. We use high performance liquid chromatography (HPLC) to show definitively that these porphyrins are uroporphyrin I and uroporphyrin III. Evidence from confocal microscopy analyses shows that the distribution of porphyrin pigments corresponds to the striking pink-red of C. pharaonius shells, as well as pink-red dots and lines on the early whorls of C. margaritarius and yellow-brown colour of later whorls. Additional HPLC results suggest that eumelanin is likely responsible for black spots. We refer to the two differently coloured porphyrin pigments as trochopuniceus (pink-red) and trochoxouthos (yellow-brown) in order to distinguish between them. Trochopuniceus and trochoxouthos were not found in the shell of a third species of the same superfamily, Calliostoma zizyphinum, despite its superficially similar colouration, suggesting that this species has different shell pigments. These findings have important implications for the study of colour and pattern in molluscs specifically, but in other taxa more generally, since this study shows that homology of visible colour cannot be assumed without identification of pigments.

Remote sensing of chlorophyll concentrations in the northern Gulf of Mexico

NASA Technical Reports Server (NTRS)

Trees, Charles C.; El-Sayed, Sayed Z.

1986-01-01

During a 17 month period (November 1978 - March 1980), phytoplankton pigment concentrations were remotely sensed in the northern Gulf of Mexico using the Coastal Zone Color Scanner (CZCS). A total of 29 CZCS orbits were processed into pigment (chlorophyll a + phaeopigments) images and then geometrically warped to a Mercator projection. A correction factor of 1.67 was applied to the pigment concentrations to correct for the tendency of the standard fluorometric method to underestimate chlorophyll a concentrations. The spatial and temporal distributions of pigment fronts were quite variable during this time series. Constant features observed throughout the pigment imagery were the entrainment of coastal waters offshore. The most extensive entrainments occurred during intrusions of the Loop Current. For the 17 month survey, the mean HPLC-corrected pigment concentration was 3.30 + or - 1.45 mg/cu m.

Interrelationships Between Maternal Carotenoid Status and Newborn Infant Macular Pigment Optical Density and Carotenoid Status

PubMed Central

Henriksen, Bradley S.; Chan, Gary; Hoffman, Robert O.; Sharifzadeh, Mohsen; Ermakov, Igor V.; Gellermann, Werner; Bernstein, Paul S.

2013-01-01

Purpose. Deposition of the macular pigment carotenoids lutein and zeaxanthin in the human retina occurs early in life. In this study, we examined the interrelationships of maternal carotenoid status and newborn infant macular pigment levels and systemic carotenoid status. As a secondary measure, we also evaluated the effects of intrauterine growth restriction (IUGR) on carotenoid status in term newborn infants. Methods. We measured mother and infant skin carotenoids using resonance Raman spectroscopy (RRS), serum carotenoids by HPLC, and mother breast milk carotenoids by HPLC. We measured infant macular pigment levels using noninvasive blue light reflectometry. Results. We enrolled 30 healthy term infants, their mothers, and 10 IUGR infants and their mothers. A subset of 16 infants was imaged for macular pigment optical density (MPOD). Infant serum zeaxanthin levels correlated with MPOD (r = 0.68, P = 0.007). Mother serum zeaxanthin levels correlated with infant MPOD (r = 0.59, P = 0.032). Infant and mother serum lutein did not correlate with MPOD. Mother–infant correlations were found for total serum carotenoids (r = 0.42, P = 0.020) and skin carotenoids (r = 0.48, P = 0.001). No difference was seen between IUGR infants and controls in total serum or skin carotenoids. Mothers of IUGR infants had lower total serum carotenoids (P = 0.019) and breast milk carotenoids than controls (P = 0.006). Conclusions. Our findings suggest that maternal zeaxanthin status may play a more important role than lutein status in macular pigment deposition in utero. Controlled trials are needed to determine whether maternal zeaxanthin prenatal supplementation can raise infant macular pigment levels and/or improve ocular function. PMID:23838770

Pigment compositions are linked to the habitat types in dinoflagellates.

PubMed

Yamada, Norico; Tanaka, Ayumi; Horiguchi, Takeo

2015-11-01

Compared to planktonic species, there is little known about the ecology, physiology, and existence of benthic dinoflagellates living in sandy beach or seafloor environments. In a previous study, we discovered 13(2),17(3)-cyclopheophorbide a enol (cPPB-aE) from sand-dwelling benthic dinoflagellates. This enol had never been detected in phytoplankton despite the fact that it is a chlorophyll a catabolite. We speculated from this discovery that habitat selection might be linked to pigment compositions in dinoflagellates. To test the hypothesis of habitat selection linking to pigment compositions, we conducted extensive analysis of pigments with high performance liquid chromatography (HPLC) for 40 species using 45 strains of dinoflagellates including three habitat types; sand-dwelling benthic forms, tidal pool inhabitants and planktonic species. The 40 dinoflagellates are also able to be distinguished into two types based on their chloroplast origins; red alga-derived secondary chloroplasts and diatom-derived tertiary ones. By plotting the pigments profiles onto three habitats, we noticed that twelve pigments including cPPB-aE were found to occur only in benthic sand-dwelling species of red alga-derived type. The similar tendency was also observed in dinoflagellates with diatom-derived chloroplasts, i.e. additional sixteen pigments including chl c 3 were found only in sand-dwelling forms. This is the first report of the occurrence of chl c 3 in dinoflagellates with diatom-derived chloroplasts. These results clarify that far greater diversity of pigments are produced by the dinoflagellates living in sand regardless of chloroplast types relative to those of planktonic and tidal pool forms. Dinoflagellates seem to produce a part of their pigments in response to their habitats.

Pigment from Streptomyces bellus MSA1 isolated from marine sediments

NASA Astrophysics Data System (ADS)

Srinivasan, M.; Merlyn Keziah, S.; Hemalatha, M.; Subathra Devi, C.

2017-11-01

The existing study is purposeful on the intracellular pigment extraction from actinomycetes isolated from Kovalam Beach regions of Chennai, Tamil Nadu, India. Only one actinobacterial isolate showed pigmented growth out of total 4 isolates. Ethyl acetate as the solvent was used in cell disruption technique for the extraction of intracellular pigments. UV-Visible spectrophotometry, FT-IR spectroscopy, HPLC and GC-MS were used for the partial characterization of the pigment. The extracted pigment was applied for the preparation of lip balm and assessing its textile dyeing property. In addition, the extracted pigment was analysed for antioxidant, antibacterial activity, MTT assay and haemolytic activity. On optimization, dextrose and maltose were the best carbon sources. The finest nitrogen sources were found to be casein and peptone. The optimum temperature range was 35°C -40°C and optimal pH was found to be between 6.0 and 8.0. The obtained results showed potent antioxidant activity and found to be non-toxic to human erythrocytes.

Hemisynthesis and structural and chromatic characterization of delphinidin 3-O-glucoside-vescalagin hybrid pigments.

PubMed

García-Estévez, Ignacio; Jacquet, Rémi; Alcalde-Eon, Cristina; Rivas-Gonzalo, Julián C; Escribano-Bailón, M Teresa; Quideau, Stéphane

2013-11-27

During red wine maturation in the presence of oak wood, reactions involving anthocyanins and ellagitannins might affect wine organoleptic properties such as color and astringency. In this work, the condensation reaction between myrtillin (delphinidin 3-O-glucoside) and vescalagin has been performed to determine the behavior of this anthocyanin in this kind of reaction and to assess the possible impact of such a reaction in wine color modulation. Two different hybrid pigments have been hemisynthetized and characterized by HPLC-DAD-MS and NMR spectroscopy. These pigments have been identified as 1-deoxyvescalagin-(1β→8)-myrtillin (major) and 1-deoxyvescalagin-(1β→6)-myrtillin (minor). The minor pigment could be formed both by the condensation reaction and by a regioisomerization process from the major pigment. Moreover, the chromatic properties of these pigments have been studied and compared to those of myrtillin. The hybrid pigments showed an important bathochromic shift (ca. 20 nm) in the maximum absorbance wavelength and lower molar absorption coefficients.

The determination and quantification of photosynthetic pigments by reverse phase high-performance liquid chromatography, thin-layer chromatography, and spectrophotometry.

PubMed

Pocock, Tessa; Król, Marianna; Huner, Norman P A

2004-01-01

Chorophylls and carotenoids are functionally important pigment molecules in photosynthetic organisms. Methods for the determination of chlorophylls a and b, beta-carotene, neoxanthin, and the pigments that are involved in photoprotective cycles such as the xanthophylls are discussed. These cycles involve the reversible de-epoxidation of violaxanthin into antheraxanthin and zeaxanthin, as well as the reversible de-epoxidation of lutein-5,6-epoxide into lutein. This chapter describes pigment extraction procedures from higher plants and green algae. Methods for the determination and quantification using high-performance liquid chromatograpy (HPLC) are described as well as methods for the separation and purification of pigments for use as standards using thin-layer chromatography (TLC). In addition, several spectrophotometric methods for the quantification of chlorophylls a and b are described.

Recovery of astaxanthin from discharged wastewater during the production of chitin

NASA Astrophysics Data System (ADS)

Chen, Xiaolin; Yang, Shengfeng; Xing, Ronge; Yu, Huahua; Liu, Song; Li, Pengcheng

2012-06-01

In this paper, studies were carried out to extract astaxanthin from discharged wastewater during the production of chitin and to reveal the scavenging effect of the obtained pigment on 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical. Different ratios of dichloromethane/methanol (V/V) were used to extract astaxanthin. When the ratio of dichloromethane/methanol was 2:8 and the ratio between the mixed organic solvent (dichloromethane/methanol, 2:8, V/V) and wastewater was 1:1, the highest yield of pigment was obtained (8.4 mg/50 mL). The concentration of free astaxanthin in the obtained pigment analyzed by HPLC was 30.02%. The obtained pigment possessed strong scavenging ability on DPPH radical and IC50 was 0.84mg/ml.

High-Performance Liquid Chromatography (HPLC) Measurements of Phytoplankton Pigment Distributions of Ocean Waters

DTIC Science & Technology

1988-11-01

coccolithophorids 19. ABSTRACT (CanMyw on rviosfe Inhcesway aM den*t byblock nmber) Until the application of high-performance liquid chromatography (HPLC) to... phycocyanin , has a maximum 0 01 absorption peak. The spectra for the 008 chlorophyll degradation products (chlo- 0.06 rophyllides, phaeophorbides and...phaeo- phytins) which are not shown in Figure z I have similar absorption maxima as their associated chlorophylls, 002 , Until the application of high

Using empirical orthogonal functions from remote sensing reflectance spectra to predict various phytoplankton pigment concentrations in the Eastern Tropical Atlantic

NASA Astrophysics Data System (ADS)

Bracher, Astrid; Taylor, Bettina; Taylor, Marc; Steinmetz, Francois; Dinter, Tilman; Röttgers, Rüdiger

2014-05-01

Phytoplankton pigments play a major role in photosynthesis and photoprotection. Their composition and abundance give information on characteristics of a phytoplankton community in respect to its acclimation to light, overall biomass and composition of major phytoplankton groups. Most phytoplankton pigments can be measured by applying HPLC techniques to filtered water samples. This method like other mathods analysing water samples in the laboratory is time consuming and therefore only a limited number of samples can be obtained. In order to obtain information on phytoplankton pigment composition with a better temporal and spatial composition, the rationale was to develop a method to get from continuous optical measurements pigment concentrations. We have used remote sensing reflectances (RRS) derived from ship-based hyper-spectral underwater radiometric and from satellite MERIS measurements (using the POLYMER algorithm developed by Steinmetz et al. 2011), sampled in the Eastern Tropical Atlantic, to predict the water surface concentration of various pigments or pigment groups in this area. A statistical model based on Empirical Orthogonal Function (EOF) analysis of these RRS spectra was developed. Then subsequently linear models with measured (collocated) pigment concentrations as the response variable and EOF loadings as predictor variables were constructed. The model results, which have been verified by cross validation, show that from the ship-based RRS measurements the surface concentrations of a suite of pigments and pigment groups can be well predicted, even when only a multi-spectral resolution of RRS data is chosen. Based on the MERIS reflectance data, only concentrations of total chlorophyll-a (chl-a), monovinyl-chl-a and the groups of photoprotective and photosynthetic carotenoids can be obtained with high quality. The model constructed on the satellite reflectances as input was also applied to one month of MERIS POLYMER data to predict for the whole Eastern Tropical Atlantic area the concentration of those pigments. Finally, the potential, limitations and future perspectives for the application of our generic method are discussed.

Cryptic color change in a crab spider (Misumena vatia): identification and quantification of precursors and ommochrome pigments by HPLC.

PubMed

Riou, Mickaël; Christidès, Jean-Philippe

2010-04-01

Mimicry is used widely by arthropods to survive in a hostile environment. Often mimicry is associated with the production of chemical compounds such as pigments. In crab spiders, the change of color is based on a complex physiological process that still is not understood. The aim of this study was to identify and quantify the ommochrome pigments and precursors responsible for the color change in the mimetic crab spider Misumena vatia (Thomisidae). A modified high performance reverse phase ion-pair chromatography technique enabled us to separate and quantify the ommochrome pigments, their precursors, and related metabolites in individual spiders. Compounds such as tryptophan, kynurenine, and kynurenic acid occurred only or mainly in white crab spiders. In contrast, compounds such as 3-hydroxy-kynurenine, xanthommatin, and ommatin D occurred only or mainly in yellow crab spiders. Factor analysis ranked the different color forms in accordance with their metabolites. The biochemical results enabled us to associate the different phases of formation of pigment granules with specific metabolites. Yellow crab spiders contain many unknown ommochrome-like compounds not present in white crab spiders. We also found large quantities of decarboxylated xanthommatin, whose role as precursor of new pathways in ommochrome synthesis needs to be assessed. The catabolism of ommochromes, a process occurring when spiders revert from yellow to white, warrants further study.

A novel red pigment from marine Arthrobacter sp. G20 with specific anticancer activity.

PubMed

Afra, S; Makhdoumi, A; Matin, M M; Feizy, J

2017-11-01

Bacterial pigments are promising compounds in the prevention and treatment of various cancers. In the current study, the antioxidant, cytotoxic and antimicrobial effects of a red pigment obtained from a marine bacterial strain were investigated. Optimization of the pigment production by the marine strain was conducted using the one-factor-at-a-time approach. Chemical identification of the pigment was achieved by UV-visible, FTIR and HPLC analyses. The biological activities of the pigment were evaluated by DPPH, MTT and microbroth dilution assays. The strain was identified as Arthrobacter, and its pigment was related to carotenoids. The EC 50 antioxidant activity of the pigment was evaluated as 4·5 mg ml -1 . It showed moderate anticancer effects on an oesophageal cancer cell line, KYSE30, while no inhibition was observed on normal HDF (human dermal fibroblasts) cells. The pigment had no antibacterial effects on the four tested strains. The antitumour activity of a carotenoid-related pigment from Arthrobacter sp. was reported for the first time. Marine environments are interesting sources for the identification of novel bioproducts. The identification of carotenoid pigments from marine bacteria with remarkable antioxidant and anticancer activities would result in better insights into the potential pharmaceutical applications of carotenoids and marine environments. © 2017 The Society for Applied Microbiology.

CZE separation of strawberry anthocyanins with acidic buffer and comparison with HPLC.

PubMed

Comandini, Patrizia; Blanda, Giampaolo; Cardinali, Andrea; Cerretani, Lorenzo; Bendini, Alessandra; Caboni, Maria Fiorenza

2008-10-01

Anthocyanins, the major colourants of strawberries, are polar pigments that are positively charged at low pH. Herein, we have assessed a new analytical method for the separation of anthocyanins using CZE. Acidic buffer solutions (pH <2) were employed in order to maintain pigments in the cation flavylium form and achieve high molar absorptivity at 510 nm. These spectral properties enabled us to identify strawberry anthocyanins in a preliminary stage by detection in the visible range, although the method was optimised at 280 nm to obtain the best S/N. The effects of buffer composition highlighted the necessity of adding an organic modifier to the running buffer to obtain a suitable separation. The electrophoretic method permitted the separation of the three main anthocyanins of strawberry extracts, namely pelargonidin 3-glucoside (Pg-glu), pelargonidin 3-rutinoside and cyanidin 3-glucoside. The electrophoretic results, expressed as retention time and separation efficiency of the major anthocyanin (Pg-glu), were compared to those achieved in HPLC, the analytical technique traditionally used for the investigation of anthocyanins in vegetable matrix. The content of Pg-glu in strawberries (cv. Camarosa), calculated with HPCE and HPLC methods, resulted respectively in 11.41 mg/L and 11.37 mg/L.

The effect of dietary carotenoid access on sexual dichromatism and plumage pigment composition in the American goldfinch.

PubMed

McGraw, K J; Hill, G E; Stradi, R; Parker, R S

2002-02-01

We investigated potential dietary and biochemical bases for carotenoid-based sexual dichromatism in American goldfinches (Carduelis tristis). Captive male and female finches were given access to the same type and amount of carotenoid pigments in the diet during their nuptial molt to assess differences in the degree to which the two sexes incorporated ingested pigments into their plumage. When birds were fed a uniform, plain-seed diet, or one that was supplemented with the red carotenoid canthaxanthin, we found that males grew more colorful plumage than females. HPLC analyses of feather pigments revealed that male finches incorporated a higher concentration of carotenoids into their pigmented feathers than females. Compared to females, males also deposited significantly more canary xanthophyll B into feathers when fed a plain-seed diet and a greater concentration and proportion of canthaxanthin when fed a carotenoid-supplemented diet. These results indicate that sex-specific expression of carotenoid pigmentation in American goldfinches may be affected by the means by which males and females physiologically utilize (e.g. absorb, transport, metabolize, deposit) carotenoid pigments available to them in the diet.

Ocean Optics Protocols for Satellite Ocean Color Sensor Validation. Revised

NASA Technical Reports Server (NTRS)

Fargion, Giulietta S.; Mueller, James L.

2000-01-01

The document stipulates protocols for measuring bio-optical and radiometric data for the Sensor Intercomparison and Merger for Biological and Interdisciplinary Oceanic Studies (SIMBIOS) Project activities and algorithm development. This document supersedes the earlier version (Mueller and Austin 1995) published as Volume 25 in the SeaWiFS Technical Report Series. This document marks a significant departure from, and improvement on, theformat and content of Mueller and Austin (1995). The authorship of the protocols has been greatly broadened to include experts specializing in some key areas. New chapters have been added to provide detailed and comprehensive protocols for stability monitoring of radiometers using portable sources, abovewater measurements of remote-sensing reflectance, spectral absorption measurements for discrete water samples, HPLC pigment analysis and fluorometric pigment analysis. Protocols were included in Mueller and Austin (1995) for each of these areas, but the new treatment makes significant advances in each topic area. There are also new chapters prescribing protocols for calibration of sun photometers and sky radiance sensors, sun photometer and sky radiance measurements and analysis, and data archival. These topic areas were barely mentioned in Mueller and Austin (1995).

Applications of Electromigration Techniques: Applications of Electromigration Techniques in Food Analysis

NASA Astrophysics Data System (ADS)

Wieczorek, Piotr; Ligor, Magdalena; Buszewski, Bogusław

Electromigration techniques, including capillary electrophoresis (CE), are widely used for separation and identification of compounds present in food products. These techniques may also be considered as alternate and complementary with respect to commonly used analytical techniques, such as high-performance liquid chromatography (HPLC), or gas chromatography (GC). Applications of CE concern the determination of high-molecular compounds, like polyphenols, including flavonoids, pigments, vitamins, food additives (preservatives, antioxidants, sweeteners, artificial pigments) are presented. Also, the method developed for the determination of proteins and peptides composed of amino acids, which are basic components of food products, are studied. Other substances such as carbohydrates, nucleic acids, biogenic amines, natural toxins, and other contaminations including pesticides and antibiotics are discussed. The possibility of CE application in food control laboratories, where analysis of the composition of food and food products are conducted, is of great importance. CE technique may be used during the control of technological processes in the food industry and for the identification of numerous compounds present in food. Due to the numerous advantages of the CE technique it is successfully used in routine food analysis.

Ion-pair high-speed countercurrent chromatography in fractionation of a high-molecular weight variation of acyl-oligosaccharide linked betacyanins from purple bracts of Bougainvillea glabra.

PubMed

Wybraniec, Sławomir; Jerz, Gerold; Gebers, Nadine; Winterhalter, Peter

2010-02-15

The natural pigment composition of purple bracts of Bougainvillea glabra (Nyctaginaceae) consists of a highly complex mixture of betacyanins solely differing by the substitution with a variety of acyl-oligoglycoside units. This study was focused on a two-dimensional chromatography approach, a combination of preparative high-speed countercurrent chromatography (HSCCC) and analytical C18-HPLC with ESI-DAD-MS/MS detection which finally enabled a more detailed view into the pigment profile and elucidated the existence of an overwhelming amount of varying betacyanin structures occurring in Bougainvillea bracts. The detected molecular weights of the pigments reached so far unknown high values and ranged up to maximum values of 1653 and 1683 Da for the largest molecules due to oligosaccharide linkage and multiple acyl substitutions. The preparative IP-HSCCC separation yielded 15 complex fractions containing betacyanins of enhanced polarity as well as structures with highly increased lipophilicity. Betacyanin structures extended by large oligosaccharide chains with bigger number of glycoside units and also carrying a reduced number of hydroxycinnamic acid substitutions were characteristic for polar pigments occurring mainly in the early eluting CCC fractions. IP-HSCCC was proven to be extremely effective for fractionating this complex crude betalain pigment extract into more defined 'polarity-windows'. Structural analysis by analytical LC-ESI-MS/MS in the positive ionization mode detected a total sum of 146 different betacyanin pigments in the CCC fractions of reduced complexity. Copyright 2010 Elsevier B.V. All rights reserved.

Monochromatic green light induces an aberrant accumulation of geranylgeranyled chlorophylls in plants.

PubMed

Materová, Zuzana; Sobotka, Roman; Zdvihalová, Barbora; Oravec, Michal; Nezval, Jakub; Karlický, Václav; Vrábl, Daniel; Štroch, Michal; Špunda, Vladimír

2017-07-01

Light quality is an important environmental factor affecting the biosynthesis of photosynthetic pigments whose production seems to be affected not only quantitatively but also qualitatively. In this work, we set out to identify unusual pigment detected in leaves of barley (Hordeum vulgare L.) and explain its presence in plants grown under monochromatic green light (GL; 500-590 nm). The chromatographic analysis (HPLC-DAD) revealed that a peak belonging to this unknown pigment is eluted between chlorophyll (Chl) a and b. This pigment exhibited the same absorption spectrum and fluorescence excitation and emission spectra as Chl a. It was negligible in control plants cultivated under white light of the same irradiance (photosynthetic photon flux density of 240 μmol m -2  s -1 ). Mass spectrometry analysis of this pigment (ions m/z = 889 [M-H] - ; m/z = 949 [M+acetic acid-H] - ) indicates that it is Chl a with a tetrahydrogengeranylgeraniol side chain (containing two double bonds in a phytyl side chain; Chl a THGG ), which is an intermediate in Chl a synthesis. In plants grown under GL, the proportion of Chl a THGG to total Chl content rose to approximately 8% and 16% after 7 and 14 days of cultivation, respectively. Surprisingly, plants cultivated under GL exhibited drastically increased concentration of the enzyme geranylgeranyl reductase, which is responsible for the reduction of phytyl chain double bonds in the Chl synthesis pathway. This indicates impaired activity of this enzyme in GL-grown plants. A similar effect of GL on Chl synthesis was observed for distinct higher plant species. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

WATER COLUMN DATA AND SPECTRAL IRRADIANCE MODEL

EPA Science Inventory

Water samples collected monthly, for 18 months, from six sites in the Laguna Madre were analyzed to identify and quantify phytopigments using High Performance Liquid Chromatography (HPLC). In addition, water column pigment and nutrient data were acquired at 12 stations in Upper ...

Estimation of color of durum wheat. Comparison of WSB, HPLC, and reflectance colorimeter measurements.

PubMed

Fratianni, Alessandra; Irano, Mario; Panfili, Gianfranco; Acquistucci, Rita

2005-04-06

Color is an important parameter involved in the definition of semolina and pasta quality. This character is mainly due to natural pigments (carotenoids) that are present at different levels in cereals and cereal products, due to botanical origin, growing conditions, distribution in the kernel, and technological processes. In food industries, color measurements are usually performed by means of automatic instruments that are rapid and safe, as alternatives to the chemical extraction methods. In this study, automatic measurements (CIE, color-space system L, a, b), water-saturated butanol (WSB), and HPLC determinations have been applied to evaluate the carotenoid content in whole meals and respective semolina samples produced from wheat cultivated in the years 2001 and 2002. In whole meals, total carotenoids, determined by HPLC, were about 3.0 microg/g (2001) and 3.5 microg/g (2002) calculated on dry weight (dw) and about 3.0 and 3.2 microg/g dw in corresponding semolina samples. The b values for the same period were 19.78 and 15.75, respectively, in raw materials and 20.03-21.67 in semolina. Results have confirmed lutein and beta-carotene as the main components mainly responsible for the yellow color in wheat grains. The ability of the index b to express natural dyeing was dependent on sample characteristics as demonstrated by the relationships found between this index and pigments, although the best correlation resulted between HPLC and WSB.

Field Spectroscopy And Spectral Analysis Of Caribbean Scleractinian Reef Corals And Related Benthic Biota

NASA Astrophysics Data System (ADS)

Torres-Perez, J. L.; Guild, L. S.; Armstrong, R.; Corredor, J. E.; Polanco, R.; Zuluaga-Montero, A. B.

2013-05-01

Coral reefs are highly heterogenic ecosystems with a plethora of photosynthetic organisms forming most of the benthic communities. Usually coral reef benthos is a composite of reef corals, different groups of algae, seagrasses, sandy bottoms, dead rubble, and even mangrove forests living in a relatively small area. The remote characterization of these important tropical ecosystems represents a challenge to scientists, particularly due to the similarity of the spectral signatures among some of these components. As such, we examined the similarities and differences between nine Scleractinian Caribbean shallow-water reef corals' spectral reflectance curves. Samples were also collected from each species for pigment analysis using High Performance Liquid Chromatography (HPLC). Reflectance curves were obtained with the aid of a GER-1500 hand-held field spectroradiometer enclosed in an underwater housing. Our findings showed that even though most of the pigmentation was directly related to the relationship of corals with their symbiotic dinoflagellates (zooxanthellae), the presence of other endolithic photosynthetic organisms can also contribute to the light absorption of corals and, hence, the reflectance spectra of each species. Also, the relative contribution of chlorophylls vs. carotenes or xanthophylls depends on the coral species with some species relying more on Chlorophyll a and other species relying on Chlorophyl c2 and Peridinin with a small Chlorophyll a component. Pigments associated with the xanthophyll cycle of dinoflagellates (Diadinoxanthin and Diatoxanthin) were detected in most species. Pigments typical of endolithic organisms such as Zeaxanthin, Fucoxanthin, Violaxanthin and Siphonaxanthin were also detected in some coral species. The influence of major pigments on the reflectance curve was evidenced with a 2nd derivative analysis. This could be used to discriminate among most species. Further, an analysis of the integration of the area under the reflectance curve in the photosynthetically active radiation (PAR; 400-700nm) yielded an inverse relationship with the total pigment concentration with an up to 97% confidence level. Corals were distinguished from seagrasses and other benthic components based on their reflectance and differences in curve inflection peaks. Special care needs to be taken when characterizing sandy bottoms as they are influenced by the presence of photosynthetic microbiota as reflected in their reflectance curves. The use of this integration is proposed as a novel non-invasive method to predict pigment changes in reef corals aimed to monitor their health in the present climate change scenario.

Diversity of Polyhydroxynaphthoquinone Pigments in North Pacific Sea Urchins.

PubMed

Vasileva, Elena A; Mishchenko, Natalia P; Fedoreyev, Sergey A

2017-09-01

Using high-performance liquid chromatography with diode-array detection and mass spectrometry (HPLC-DAD/MS) we investigated the composition of polyhydroxynaphthoquinone (PHNQ) pigments from sea urchins Strongylocentrotus pallidus, St. polyacanthus, St. droebachiensis, Brisaster latifrons and Echinarachnius parma, collected in the Sea of Okhotsk and the Bering Sea. Identification of PHNQ pigments from sea urchins St. polyacanthus, B. latifrons, and E. parma was performed for the first time. Among the usual PHNQ pigments, mono- and dimethoxy derivatives of spinochrome E, not previously found in other sea urchins, were discovered in St. polyacanthus and St. droebachiensis. In St. droebachiensis, two monomethoxy derivatives of echinochrome A were detected, isolated previously from only tropical sea urchins. It was found that the composition and total content of pigments of St. droebachiensis depends on the collection area of the sea urchins and its depth and varies from 88 to 331 μg/g of dry shells. Sea urchins St. pallidus, B. latifrons and E. parma had average values for PHNQ pigment content, approximately 30 μg/g, and St. polyacanthus had a low PHNQ content, 13 μg/g. © 2017 Wiley-VHCA AG, Zurich, Switzerland.

Spatial-temporal distribution of phytoplankton pigments in relation to nutrient status in Jiaozhou Bay, China

NASA Astrophysics Data System (ADS)

Yao, Peng; Yu, Zhigang; Deng, Chunmei; Liu, Shuxia; Zhen, Yu

2010-10-01

We conducted studies of phytoplankton and hydrological variables in a semi-enclosed bay in northern China to understand the spatial-temporal variability and relationship between these variables. Samples were collected during seven cruises in Jiaozhou Bay from November 2003 to October 2004, and were analyzed for temperature, nutrients and phytoplankton pigments. Pigments from eight possible phytoplankton classes (Diatoms, Dinoflagellates, Chlorophyceae, Prasinophyceae, Chrysophyceae, Haptophyceae, Cryptophyceae and Caynophyceae) were detected in surface water by high performance liquid chromatography (HPLC). Phytoplankton pigment and nutrient concentrations in Jiaozhou Bay were spatially and temporally variable, and most of them were highest in the northern and eastern parts of the sampling regions in spring (May) and summer (August), close to areas of shellfish culturing, river estuaries, dense population and high industrialization, reflecting human activities. Chlorophyll a was recorded in all samples, with an annual mean concentration of 1.892 μg L -1, and fucoxanthin was the most abundant accessory pigment, with a mean concentration of 0.791 μg L -1. The highest concentrations of chlorophyll a (15.299 μg L -1) and fucoxanthin (9.417 μg L -1) were observed in May 2004 at the station close to the Qingdao Xiaogang Ferry, indicating a spring bloom of Diatoms in this area. Although chlorophyll a and other biomarker pigments showed significant correlations, none of them showed strong correlations with temperature and nutrients, suggesting an apparent de-coupling between the pigments and these hydrological variables. The nutrient composition and phytoplankton community composition of Jiaozhou Bay have changed significantly in the past several decades, reflecting the increasing nutrient concentrations and decline of phytoplankton cell abundance. The unchanged total chlorophyll a levels indicated that smaller species have filled the niche vacated by the larger species in Jiaozhou Bay, as revealed by our biomarker pigment analysis.

Evaluation of processing effects on anthocyanin content and colour modifications of blueberry (Vaccinium spp.) extracts: Comparison between HPLC-DAD and CIELAB analyses.

PubMed

Cesa, Stefania; Carradori, Simone; Bellagamba, Giuseppe; Locatelli, Marcello; Casadei, Maria Antonietta; Masci, Alessandra; Paolicelli, Patrizia

2017-10-01

Colour is the first organoleptic property that consumers appreciate of a foodstuff. In blueberry (Vaccinium spp.) fruits, the anthocyanins are the principal pigments determining the colour as well as many of the beneficial effects attributed to this functional food. Commercial blueberry-derived products represent important sources of these healthy molecules all year round. In this study, blueberries were produced into purees comparing two homogenization methods and further heated following different thermal treatments. All the supernatants of the homogenates were monitored for pH. Then, the hydroalcoholic extracts of the same samples were characterized by CIELAB and HPLC-DAD analyses. These analytical techniques provide complementary information on fruit pigments content as a whole and on quali-quantitative profile of the single bioactive colorants. These data could be very interesting to know the best manufacturing procedure to prepare blueberry-derived products, well accepted by the consumers, while maintaining their healthy properties unaltered. Copyright © 2017. Published by Elsevier Ltd.

Separation and identification of anthocyanin extracted from mulberry fruit and the pigment binding properties toward human serum albumin.

PubMed

Sheng, Feng; Wang, Yuning; Zhao, Xingchen; Tian, Na; Hu, Huali; Li, Pengxia

2014-07-16

Purple pigments were isolated from mulberry extracts using preparative high-speed countercurrent chromatography (HSCCC) and identified by ESI-MS/MS and high performance liquid chromatography (HPLC) techniques. The solvent system containing methyl tert-butyl ether, 1-butanol, acetonitrile, water, and trifluoroacetic acid (10:30:10:50:0.05; %, v/v) was developed in order to separate anthocyanins with different polarities. Cyanidin 3-O-(6″-O-α-rhamnopyranosyl-β-galactopyranoside) (also known as keracyanin) is the major component present in mulberry (41.3%). Other isolated pigments are cyanidin 3-O-(6″-O-α-rhamnopyranosyl-β-glucopyranoside) and petunidin 3-O-β-glucopyranoside. The binding characteristics of keracyanin with human serum albumin (HSA) were investigated by fluorescence and circular dichroism (CD) spectroscopy. Spectroscopic analysis reveals that HSA fluorescence quenched by keracyanin follows a static mode. Binding of keracyanin to HSA mainly depends on van der Waals force or H-bonds with average binding distance of 2.82 nm. The results from synchronous fluorescence, three-dimensional fluorescence, and CD spectra show that adaptive structure rearrangement and decrease of α-helical structure occur in the presence of keracyanin.

Simultaneous determination of phenolic acids and flavonoids in rice using solid-phase extraction and RP-HPLC with photodiode array detection.

PubMed

Irakli, Maria N; Samanidou, Victoria F; Biliaderis, Costas G; Papadoyannis, Ioannis N

2012-07-01

An analytical method based on an optimized solid-phase extraction procedure and followed by high-performance liquid chromatography (HPLC) separation with diode array detection was developed and validated for the simultaneous determination of phenolic acids (gallic, protocatechuic, 4-hydroxy-benzoic, vanillic, caffeic, syringic, p-coumaric, ferulic, sinapic, and cinnamic acids), flavanols (catechin and epicatechin), flavonols (myricetin, quercetin, kaempferol, quercetin-3-O-glucoside, hyperoside, and rutin), flavones (luteolin and apigenin) and flavanones (naringenin and hesperidin) in rice flour (Oryza sativa L.). Chromatographic separation was carried out on a PerfectSil Target ODS-3 (250 mm × 4.6 mm, 3 μm) column at temperature 25°C using a mobile phase, consisting of 0.5% (v/v) acetic acid in water, methanol, and acetonitrile at a flow rate 1 mL min(-1) , under gradient elution conditions. Application of optimum extraction conditions, elaborated on both Lichrolut C(18) and Oasis HLB cartridges, have led to extraction of phenolic acids and flavonoids from rice flour with mean recoveries 84.3-113.0%. The developed method was validated in terms of linearity, accuracy, precision, stability, and sensitivity. Repeatability (n = 5) and inter-day precision (n = 4) revealed relative standard deviation (RSD) <13%. The optimized method was successfully applied to the analysis of phenolic acids and flavonoids in pigmented (red and black rice) and non-pigmented rice (brown rice) samples. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A simple subcritical chromatographic test for an extended ODS high performance liquid chromatography column classification.

PubMed

Lesellier, Eric; Tchapla, Alain

2005-12-23

This paper describes a new test designed in subcritical fluid chromatography (SFC) to compare the commercial C18 stationary phase properties. This test provides, from a single analysis of carotenoid pigments, the absolute hydrophobicity, the silanol activity and the steric separation factor of the ODS stationary phases. Both the choice of the analytical conditions and the validation of the information obtained from the chromatographic measurements are detailed. Correlations of the carotenoid test results with results obtained from other tests (Tanaka, Engelhard, Sander and Wise) performed both in SFC and HPLC are discussed. Two separation factors, calculated from the retention of carotenoid pigments used as probe, allowed to draw a first classification diagram. Columns, which present identical chromatographic behaviors are located in the same area on this diagram. This location can be related to the stationary phase properties: endcapping treatments, bonding density, linkage functionality, specific area or silica pore diameter. From the first classification, eight groups of columns are distinguished. One group of polymer coated silica, three groups of polymeric octadecyl phases, depending on the pore size and the endcapping treatment, and four groups of monomeric stationary phases. An additional classification of the four monomeric groups allows the comparison of these stationary phases inside each group by using the total hydrophobicity. One hundred and twenty-nine columns were analysed by this simple and rapid test, which allows a comparison of columns with the aim of helping along their choice in HPLC.

Presence of exclusively bacteriochlorophyll-c containing substrain in the culture of green sulfur photosynthetic bacterium Chlorobium vibrioforme strain NCIB 8327 producing bacteriochlorophyll-d.

PubMed

Saga, Yoshitaka; Oh-oka, Hirozo; Hayashi, Takashi; Tamiaki, Hitoshi

2003-12-01

The light-dependent composition change of light harvesting bacteriochlorophyll(BChl)s in the present culture of a green sulfur photosynthetic bacterium Chlorobium (Chl.) vibrioforme f. sp. thiosulfatophilum strain NCIB 8327 was investigated by visible absorption spectroscopy and HPLC analyses. When the culture was repeatedly grown in liquid media under a low light condition, both the Soret and Qy absorption bands of the in vivo spectrum were shifted to longer wavelengths. Analysis of the extracted pigments by HPLC revealed that the ratio of the amount of BChl-c to that of BChl-d molecules gradually increased during repeated cultivation. In contrast, when the culture grown under a low light intensity was transferred to a high light condition and continued to be grown, the absorption bands were shifted to shorter wavelengths and the ratio of BChls-c/d decreased finally to the almost original value. Colonies were prepared on solid agar media from the liquid culture containing both BChls-c and d, which was grown under a low light intensity. Each colony obtained was found to contain either BChl-c or d, but not both of them. Two types of cells isolated in this study were derived from the same clone, judged from their genetic analyses. The variation of pigment composition in our liquid culture observed here could be ascribed to the difference of growth rates between two substrains containing BChl-c and BChl-d, respectively, depending on light conditions.

Structure of Pigment Metabolic Pathways and Their Contributions to White Tepal Color Formation of Chinese Narcissus tazetta var. chinensis cv Jinzhanyintai

PubMed Central

Yang, Jingwen; Lu, Bingguo; Jiang, Yaping; Chen, Haiyang; Hong, Yuwei; Wu, Binghua; Miao, Ying

2017-01-01

Chinese narcissus (Narcissus tazetta var. chinensis) is one of the ten traditional flowers in China and a famous bulb flower in the world flower market. However, only white color tepals are formed in mature flowers of the cultivated varieties, which constrains their applicable occasions. Unfortunately, for lack of genome information of narcissus species, the explanation of tepal color formation of Chinese narcissus is still not clear. Concerning no genome information, the application of transcriptome profile to dissect biological phenomena in plants was reported to be effective. As known, pigments are metabolites of related metabolic pathways, which dominantly decide flower color. In this study, transcriptome profile and pigment metabolite analysis methods were used in the most widely cultivated Chinese narcissus “Jinzhanyintai” to discover the structure of pigment metabolic pathways and their contributions to white tepal color formation during flower development and pigmentation processes. By using comparative KEGG pathway enrichment analysis, three pathways related to flavonoid, carotenoid and chlorophyll pigment metabolism showed significant variations. The structure of flavonoids metabolic pathway was depicted, but, due to the lack of F3ʹ5ʹH gene; the decreased expression of C4H, CHS and ANS genes; and the high expression of FLS gene, the effect of this pathway to synthesize functional anthocyanins in tepals was weak. Similarly, the expression of DXS, MCT and PSY genes in carotenoids synthesis sub-pathway was decreased, while CCD1/CCD4 genes in carotenoids degradation sub-pathway was increased; therefore, the effect of carotenoids metabolic pathway to synthesize adequate color pigments in tepals is restricted. Interestingly, genes in chlorophyll synthesis sub-pathway displayed uniform down-regulated expression, while genes in heme formation and chlorophyll breakdown sub-pathways displayed up-regulated expression, which also indicates negative regulation of chlorophyll formation. Further, content change trends of various color metabolites detected by HPLC in tepals are consistent with the additive gene expression patterns in each pathway. Therefore, all three pathways exhibit negative control of color pigments synthesis in tepals, finally resulting in the formation of white tepals. Interestingly, the content of chlorophyll was more than 10-fold higher than flavonoids and carotenoids metabolites, which indicates that chlorophyll metabolic pathway may play the major role in deciding tepal color formation of Chinese narcissus. PMID:28885552

Structure of Pigment Metabolic Pathways and Their Contributions to White Tepal Color Formation of Chinese Narcissus tazetta var. chinensis cv Jinzhanyintai.

PubMed

Ren, Yujun; Yang, Jingwen; Lu, Bingguo; Jiang, Yaping; Chen, Haiyang; Hong, Yuwei; Wu, Binghua; Miao, Ying

2017-09-08

Chinese narcissus ( Narcissus tazetta var. chinensis ) is one of the ten traditional flowers in China and a famous bulb flower in the world flower market. However, only white color tepals are formed in mature flowers of the cultivated varieties, which constrains their applicable occasions. Unfortunately, for lack of genome information of narcissus species, the explanation of tepal color formation of Chinese narcissus is still not clear. Concerning no genome information, the application of transcriptome profile to dissect biological phenomena in plants was reported to be effective. As known, pigments are metabolites of related metabolic pathways, which dominantly decide flower color. In this study, transcriptome profile and pigment metabolite analysis methods were used in the most widely cultivated Chinese narcissus "Jinzhanyintai" to discover the structure of pigment metabolic pathways and their contributions to white tepal color formation during flower development and pigmentation processes. By using comparative KEGG pathway enrichment analysis, three pathways related to flavonoid, carotenoid and chlorophyll pigment metabolism showed significant variations. The structure of flavonoids metabolic pathway was depicted, but, due to the lack of F3'5'H gene; the decreased expression of C4H , CHS and ANS genes; and the high expression of FLS gene, the effect of this pathway to synthesize functional anthocyanins in tepals was weak. Similarly, the expression of DXS , MCT and PSY genes in carotenoids synthesis sub-pathway was decreased, while CCD1 / CCD4 genes in carotenoids degradation sub-pathway was increased; therefore, the effect of carotenoids metabolic pathway to synthesize adequate color pigments in tepals is restricted. Interestingly, genes in chlorophyll synthesis sub-pathway displayed uniform down-regulated expression, while genes in heme formation and chlorophyll breakdown sub-pathways displayed up-regulated expression, which also indicates negative regulation of chlorophyll formation. Further, content change trends of various color metabolites detected by HPLC in tepals are consistent with the additive gene expression patterns in each pathway. Therefore, all three pathways exhibit negative control of color pigments synthesis in tepals, finally resulting in the formation of white tepals. Interestingly, the content of chlorophyll was more than 10-fold higher than flavonoids and carotenoids metabolites, which indicates that chlorophyll metabolic pathway may play the major role in deciding tepal color formation of Chinese narcissus.

Biomarker pigment signatures in Cochin back water system - A tropical estuary south west coast of India

NASA Astrophysics Data System (ADS)

Aneeshkumar, N.; Sujatha, C. H.

2012-03-01

Sedimentary biomarker pigments around Cochin estuary situated in the southwest coast of India were determined by HPLC. Fucoxanthin, an indicator of diatom was observed to be the most abundant carotenoid pigment in the estuary. Dinoflagellate derived carotenoid pigment peridinin was confined in the southern part of estuary and zeaxanthin pigment indicative of cyanobacteria were more found in sites influenced by anthropogenic activities. One compound having close similarity to fucoxanthin was also detected. Alloxanthin (cryptophyceae), chl b (green algae), canthaxanthin, neoxanthin, lutein and peridinin isomer were also detected by spectra and corresponding algal class were identified. The highest concentration of chl a (11.01 μg g-1) found near to the anthropogenic affected area while the lowest chl a (0.65 μg g-1) was recorded in industrial area. Degradation products of chl a, such as pheophorbide and pheophytin were observed and principal mode of mechanism of degradation were derived. Higher pheopigments content than chl a, reflects a density trapping of dead cells and early degradation of phytopigments from grazing activities.

Epiphytic pink-pigmented methylotrophic bacteria enhance germination and seedling growth of wheat (Triticum aestivum) by producing phytohormone.

PubMed

Meena, Kamlesh K; Kumar, Manish; Kalyuzhnaya, Marina G; Yandigeri, Mahesh S; Singh, Dhananjaya P; Saxena, Anil K; Arora, Dilip K

2012-05-01

Methylotrophic bacteria were isolated from the phyllosphere of different crop plants such as sugarcane, pigeonpea, mustard, potato and radish. The methylotrophic isolates were differentiated based on growth characteristics and colony morphology on methanol supplemented ammonium mineral salts medium. Amplification of the mxaF gene helped in the identification of the methylotrophic isolates as belonging to the genus Methylobacterium. Cell-free culture filtrates of these strains enhanced seed germination of wheat (Triticum aestivum) with highest values of 98.3% observed using Methylobacterium sp. (NC4). Highest values of seedling length and vigour were recorded with Methylobacterium sp. (NC28). HPLC analysis of production by bacterial strains ranged from 1.09 to 9.89 μg ml(-1) of cytokinins in the culture filtrate. Such cytokinin producing beneficial methylotrophs can be useful in developing bio-inoculants through co-inoculation of pink-pigmented facultative methylotrophs with other compatible bacterial strains, for improving plant growth and productivity, in an environment-friendly manner.

Photosynthetic pigments as biomarkers oof phytoplankton populations and processes involved in the transformation of particulate organic matter at the Biotrans site (47°N, 20°W)

NASA Astrophysics Data System (ADS)

Williams, R.; Claustre, H.

1991-03-01

Particulate samples were collected throughout the water column (0-4200 m)in June/July 1988 at the Biotrans site and their carotenoids and chlorophylls analysed by HPLC. These photosynthetic pigments were used as biomarkers to characterize the autotropiic pupolations, their utilization by heterotrophs snd sedimentation of particles out the euphotic zone. In the upper 50 m the pico- and nanophytoplankton accounted for 85% of the chlorophyll a biomass. The major pigment of the nanophytoplankton fraction was 19'-hexanolloxyfucoxanthin (prymnesiophytes), whereas the main pigment in the microphytoplankton was peridinin (dinoflagellates). The peaks in the distributions of phaeophorbide a and nanophytoplankton pigments (19'- hexanoyloxyfucoxanthin, 19'-butanoyloxyfucoxanthin, chlorophyll b,lutein and/or zeaxanthin) coincided between 75 and 100 m, which pointed to an active grazing of nanophytoplankton by zooplankton. These pigments were detected in particles >20 μm from the Double Longhurst Hardy Plankton Recorder down to 1000 m, probably as a consequence of their incorporation into sedimenting faecal material. In contrast, the vertical distributions of phaeophorbide a and peridinin (microphytoplankton pigment) did not coincide, and this carotenoid was not detected below 400 m in particles > 20μm. A vertical profile (0-4200 m) shows,at 2300 m, the presence of nanophytoplanktonic material similar in its pigment pattern and compositionto that of surface populations, suggesting fast sedimentation of Prymnesiophyte floc.

The stereochemistry of chlorophyll-c₃ from the haptophyte Emiliania huxleyi: the (13²R)-enantiomers of chlorophylls-c are exclusively selected as the photosynthetically active pigments in chromophyte algae.

PubMed

Mizoguchi, Tadashi; Kimura, Yuki; Yoshitomi, Taichi; Tamiaki, Hitoshi

2011-11-01

Chlorophyll(Chl)-c pigments in algae, diatoms and some prokaryotes are characterized by the fully conjugated porphyrin π-system as well as the acrylate residue at the 17-position. The precise structural characterization of Chl-c(3) from the haptophyte Emiliania huxleyi was performed. The conformations of the π-conjugated peripheral substituents, the 3-/8-vinyl, 7-methoxycarbonyl and 17-acrylate moieties were evaluated, in a solution, using nuclear Overhauser enhancement correlations and molecular modeling calculations. The rotation of the 17-acrylate residue was considerably restricted, whereas the other three substituents readily rotated at ambient temperature. Moreover, the stereochemistry at the 13²-position was determined by combination of chiral high-performance liquid chromatography (HPLC) with circular dichroism (CD) spectroscopy. Compared with the CD spectra of the structurally related, synthetic (13²R)- and (13²S)-protochlorophyllide(PChlide)-a, naturally occurring Chl-c₃ had exclusively the (13²R)-configuration. To elucidate this natural selection of a single enantiomer, we analyzed the three major Chl-c pigments (Chl-c₁, c₂ and c₃) in four phylogenetically distinct classes of Chl-c containing algae, i.e., heterokontophyta, dinophyta, cryptophyta and haptophyta using chiral HPLC. All the photosynthetic organisms contained only the (13²R)-enantiomerically pure Chls-c, and lacked the corresponding enantiomeric (13²S)-forms. Additionally, Chl-c₂ was found in all the organisms as the common Chl-c. These results throw a light on the biosynthesis as well as photosynthetic function of Chl-c pigments: Chl-c₂ is derived from 8-vinyl-PChlide-a by dehydrogenation of the 17-propionate to acrylate residues as generally proposed, and the (13²R)-enantiomers of Chls-c function as photosynthetically active, light-harvesting pigments together with the principal Chl-a and carotenoids. 2011 Elsevier B.V. All rights reserved.

Anti-quorum sensing activity of Pistacia atlantica against Pseudomonas aeruginosa PAO1 and identification of its bioactive compounds.

PubMed

Kordbacheh, H; Eftekhar, F; Ebrahimi, S N

2017-09-01

Pseudomonas aeruginosa is a multidrug resistant opportunistic pathogen and an important cause of nosocomial infections. Quorum-sensing (QS) is a process in which bacterial cell-cell communication can regulates production of many virulence factors including pigment formation and the ability to form biofilm which is essential for establishment of chronic infections. We examined the inhibitory effect of Pistacia atlantica (Anacardiaceae) methanolic leaf extract and its bioactive components on biofilm formation and pigment production by P. aeruginosa PAO1. Fractionation of the methanolic leaf extract was carried out using HPLC based activity profiling. Identification of the active compounds was carried out by the integrated approach of HPLC-DAD and LC-MS followed by molecular docking analysis. Pistacia atlantica crude extract at 2 and 1 mg/mL, inhibited 92% and 79% biofilm formation, respectively. Minimum biofilm inhibitory concentration (MBIC) determined by microbroth dilution was 0.25 mg/mL with 39% inhibition. Pyocyanin production measured by spectrophotometry showed 100% and 83% inhibition at 2 and 1 mg/mL and minimum inhibitory concentration (MIC) was 0.5 mg/mL with 40% inhibition. Four active HPLC fractions (11, 15, 16 and 19) showed MBIC values of 0.06, 0.16, 0.10, 0.15 mg/mL, and MICs for pyocyanin production of 0.49, 0.31, 0.76, >0.30 mg/mL, respectively. The active compounds were identified as rutin (1), myricetin, 3-O-rutinoside (2) and kaempferol-3-O-rutinoside (4), all belonging to the flavonoid family. Molecular docking simulation of the active compounds showed that all had high affinity for LasR protein which is an important quorum-sensing signal receptor. The results of this study suggest that the active components of P. atlantica have high anti-QS activities and may have the potential for treatment of chronic infections caused by Pseudomonas aeruginosa. Copyright © 2017 Elsevier Ltd. All rights reserved.

Production and New Extraction Method of Polyketide Red Pigments Produced by Ascomycetous Fungi from Terrestrial and Marine Habitats

PubMed Central

Lebeau, Juliana; Venkatachalam, Mekala; Fouillaud, Mireille; Petit, Thomas; Vinale, Francesco; Dufossé, Laurent; Caro, Yanis

2017-01-01

The use of ascomycetous fungi as pigment producers opens the way to an alternative to synthetic dyes, especially in the red-dye industries, which have very few natural pigment alternatives. The present paper aimed to bio-prospect and screen out 15 selected ascomycetous fungal strains, originating from terrestrial and marine habitats belonging to seven different genera (Penicillium, Talaromyces, Fusarium, Aspergillus, Trichoderma, Dreschlera, and Paecilomyces). We identified four strains, Penicillium purpurogenum rubisclerotium, Fusarium oxysporum, marine strains identified as Talaromyces spp., and Trichoderma atroviride, as potential red pigment producers. The extraction of the pigments is a crucial step, whereby the qualitative and quantitative compositions of each fungal extract need to be respected for reliable identification, as well as preserving bioactivity. Furthermore, there is a growing demand for more sustainable and cost-effective extraction methods. Therefore, a pressurized liquid extraction technique was carried out in this study, allowing a greener and faster extraction step of the pigments, while preserving their chemical structures and bioactivities in comparison to conventional extraction processes. The protocol was illustrated with the production of pigment extracts from P. purpurogenum rubisclerotium and Talaromyces spp. Extracts were analyzed by high-performance liquid-chromatography combined with photodiode array-detection (HPLC-DAD) and high-resolution mass spectrometry (UHPLC-HRMS). The more promising strain was the isolate Talaromyces spp. of marine origin. The main polyketide pigment produced by this strain has been characterized as N-threoninerubropunctamine, a non-toxic red Monascus-like azaphilone pigment. PMID:29371552

Increase in Dominance of Eukaryote Over Prokaryote Phytoplankton Biomass Between the Surface and the Deep Chlorophyll Maximum in the Summertime Western North Atlantic Ocean

NASA Astrophysics Data System (ADS)

Vaillancourt, R. D.; Lance, V. P.; Hargreaves, B. R.; Marra, J. F.

2016-02-01

We report a general increase in the dominance of eukaryotes phytoplankton between the surface and the deep chlorophyll maximum (DCM) depths in the western North Atlantic Ocean along a transect between Bermuda (BATS) and the New England continental shelf sea during the summer 2007 & 2008. At each of 40 stations HPLC pigment concentrations were determined from 6 -10 depths from the surface to near or below the base of the euphotic zone. The community composition was determined using CHEMTAX (Mackey et al. 1996) using marker pigment ratios for varying light regimes published in Higgins et al (2011) and from our own monocultures. Cluster analysis was used to partition the dataset into five distinct regional groups to reduce the pigment ratio variability in CHEMTAX runs. Within each regional group the data were again clustered depth-wise into five to seven overlapping optical depth (OD) bins, and each OD bin was analyzed using a pigment ratio matrix ideal for that light depth range. This analysis revealed the likely presence of nine pigment classes: pelagophytes, diatoms, dinoflagellates, Synechococcus sp., Prochlorococcs sp., cryptophytes, chlorophytes, prasinophytes, and haptophytes. Partial verification of CHEMTAX results was obtained using flow cytometry cell counts coincident with samples from the BATS stations that show reasonable (according to published values) Chl a/cell values for surface and deep populations. At most locations and depths, the eukaryote haptophyte group dominated the phytoplankton biomass. In the upper optical depth the proportion of phytoplankton biomass contributed by prokaryotes was 39 (± 23)%. Deeper, between OD 1 and 2, this proportion decreased to 33 (± 17)%, between ODs 2 and 4.6 to 25 (± 15)%, and below OD 4.6, to 21 (± 17)%. Some geographic variation was observed, with the trend most pronounced in oligotrophic ocean waters and weaker in continental shelf waters.

Determination of Beet Root Betanin in Dairy Products by High-Performance Liquid Chromatography (HPLC)

ERIC Educational Resources Information Center

Gandia-Herrero, Fernando; Simon-Carrillo, Ana; Escribano, Josefa; Garcia-Carmona, Francisco

2012-01-01

The food industry uses different additives to give foods and beverages the appearance expected by the consumer. Among them, pigments of natural origin are receiving increasing attention due to safety concerns about traditional colorants and the relevance of a healthy diet. This experiment describes the quantitative determination of the…

New HPLC methods to quantitate terpenoid aldehydes in foliage of cotton (Gossypium)

USDA-ARS?s Scientific Manuscript database

The cotton plant (Gossypium) produces protective terpenoid aldehydes in lysigenous pigment glands. These terpenoids include hemigossypolone, hemigossypolone-6-methyl ether, gossypol, gossypol-6-methyl ether, gossypol-6,6'-dimethyl ether, heliocides H1, H2, H3 and H4, and heliocides B1, B2, B3 and B4...

Animal pigment bilirubin discovered in plants.

PubMed

Pirone, Cary; Quirke, J Martin E; Priestap, Horacio A; Lee, David W

2009-03-04

The bile pigment bilirubin-IXalpha is the degradative product of heme, distributed among mammals and some other vertebrates. It can be recognized as the pigment responsible for the yellow color of jaundice and healing bruises. In this paper we present the first example of the isolation of bilirubin in plants. The compound was isolated from the brilliant orange-colored arils of Strelitzia nicolai, the white bird of paradise tree, and characterized by HPLC-ESMS, UV-visible, (1)H NMR, and (13)C NMR spectroscopy, as well as comparison with an authentic standard. This discovery indicates that plant cyclic tetrapyrroles may undergo degradation by a previously unknown pathway. Preliminary analyses of related plants, including S. reginae, the bird of paradise, also revealed bilirubin in the arils and flowers, indicating that the occurrence of bilirubin is not limited to a single species or tissue type.

Determination of anthocyanins from camu-camu (Myrciaria dubia) by HPLC-PDA, HPLC-MS, and NMR.

PubMed

Zanatta, Cinthia Fernanda; Cuevas, Elyana; Bobbio, Florinda O; Winterhalter, Peter; Mercadante, Adriana Z

2005-11-30

Camu-camu [Myrciaria dubia (HBK) McVaugh] is a small fruit native to the Amazonian rain forest. Its anthocyanin profile has now been investigated for the first time. Fruits from two different regions of the São Paulo state, Brazil, were analyzed. The major anthocyanins were isolated by high-speed countercurrent chromatography. HPLC-PDA, HPLC-MS/MS, and 1H NMR were used to confirm the identity of the main anthocyanins of camu-camu. Cyanidin-3-glucoside was identified as the major pigment in the fruits from both regions, representing 89.5% in the fruits produced in Iguape and 88.0% in those from Mirandópolis, followed by the delphinidin-3-glucoside, ranging between 4.2 and 5.1%, respectively. Higher total anthocyanin contents were detected in the fruits from Iguape (54.0 +/- 25.9 mg/100 g) compared to those from Mirandópolis (30.3 +/- 6.8 mg/100 g), most likely because of the lower temperatures in the Iguape region.

Contrasting patterns of phytoplankton pigments and chemotaxonomic groups along 30°S in the subtropical South Atlantic Ocean

NASA Astrophysics Data System (ADS)

Araujo, Milton Luiz Vieira; Mendes, Carlos Rafael Borges; Tavano, Virginia Maria; Garcia, Carlos Alberto Eiras; Baringer, Molly O'Neil

2017-02-01

This work describes the spatial distribution of pigments and main taxonomic groups of phytoplankton in the biogeochemical provinces of the subtropical South Atlantic Ocean, along 30°S latitude. Seawater samples (surface to 200 m depth) were collected along 120 oceanographic stations occupied in the early austral spring of 2011, during a CLIVAR Repeat Hydrography cruise. The pigments were identified and quantified by high performance liquid chromatography (HPLC), and CHEMTAX software was used to determine the relative contributions of the main taxonomic groups to total chlorophyll a (phytoplankton biomass index). Sampling stations were grouped into three provinces: Africa, Gyre, and Brazil, corresponding to the eastern, central, and western sectors of the transect, respectively. Our results showed that both vertical and horizontal distribution patterns of pigments and taxonomic groups were mainly determined by the availability of light and/or nutrients. Photosynthetic carotenoids (PSCs), associated with small flagellates (mainly haptophytes), dominated the light-limited and nutrient-enhanced deep chlorophyll maximum (DCM) layers of both the Brazil and Gyre provinces, as well as the upwelling influenced surface waters of the Africa province. The latter showed the highest chlorophyll a values ​​(>1 mg m-3) and abundance of dinoflagellates in the coastal region. Photoprotective carotenoids (PPCs) were predominant in the nutrient-poor and well-lit surface layers of the Brazil and Gyre provinces, associated with a low content of chlorophyll a ( 0.1 mg m-3) and dominance of prokaryotes (Synechoccocus and Prochloroccocus). This study demonstrates that pigment analysis can provide a useful approach to better understand the distribution of phytoplankton communities along physical-chemical gradients in a still undersampled region of the South Atlantic Ocean.

Sedimentary pigments and nature of organic matter within the oxygen minimum zone (OMZ) of the Eastern Arabian Sea (Indian margin)

NASA Astrophysics Data System (ADS)

Rasiq, K. T.; Kurian, S.; Karapurkar, S. G.; Naqvi, S. W. A.

2016-07-01

Sedimentary pigments, carbon and nitrogen content and their stable isotopes were studied in three short cores collected from the oxygen minimum zone (OMZ) of the Eastern Arabian Sea (EAS). Nine pigments including chlorophyll a and their degradation products were quantified using High Performance Liquid Chromatography (HPLC). Astaxanthin followed by canthaxanthin and zeaxanthin were the major carotenoids detected in these cores. The total pigment concentration was high in the core collected from 500 m water depth (6.5 μgg-1) followed by 800 m (1.7 μgg-1) and 1100 m (1.1 μgg-1) depths respectively. The organic carbon did not have considerable control on sedimentary pigments preservation. Pigment degradation was comparatively high in the core collected from the 800 m site which depended not only the bottom dissolved oxygen levels, but also on the faunal activity. As reported earlier, the bottom water dissolved oxygen and presence of fauna have good control on the organic carbon accumulation and preservation at Indian margin OMZ sediments. The C/N ratios and δ13C values for all the cores conclude the marine origin of organic matter and δ15N profiles revealed signature of upwelling associated denitrification within the water column.

Study of phytoplankton group distribution in the NW African upwelling system and its relation with hydrographical parameters

NASA Astrophysics Data System (ADS)

Vega Moreno, Daura; Llerandi-García, C.; Pérez-Marrero, J.; Rueda, M. J.; Llinás, O.

2010-05-01

Phytoplankton is one important factor in biogeochemical cycles in the ocean, controlling partly the carbon cycle in the ocean. The CO2 captured by phytoplankton in shallow waters is transported to deeper layers in the ocean (Anderson, 2005). This only happens when microplankton or nanoplankton is formed due to their bigger size contributes their sinking to deeper layers. In the case of picoplankton the carbon is recycled in the same zone where is produced. The type of phytoplankton predominant can vary according to hydrographical and chemical properties present (Jeffrey et al., 2005). Pigment speciation can provide valuable ecological information in the form of classification of phytoplanktonic biomass in different groups (chlorophyta, diatoms, dinoflagellates, coccolithophorids or silicoflagellates) organized in different sizes: microphytoplankton (20-200 µm), nanophytoplankton (2-20 µm) and picophytoplankton (0.2-2 µm). There are different data processing methodologies for obtaining these classifications, one of the most accepted is the procedure taken by NASA (Hooker et al., 2005) developed by Vidussi et al. (2001), and also the use of CHEMTAX program (Mackey, 1996). It was studied pigment composition of phytoplankton and several physical and chemical properties, focused in the Northwestern African Upwelling area, including Senegal area, Mauritanian and Cape Blanc area and Morroco area. Pigment composition was analyzed by high pressure liquid chromatography and determined in different samples in the studied area, with this composition it was obtained phytoplankton classification, according to their size and to the different phytoplankton groups. These results have been related with marine biogeochemical factors presents in studied zone. References: - Anderson, T. Plankton functional type modelling: running before we can walk? Journal of Plankton Research, 2005, 27, 1073-7081 - Hooker, S.; Heukelem, L.; Thomas, C.; Claustre, H.; Ras, J.; Barlow, R.; Sessions, H.; Schlüter, L.; Perl, J.; Trees, C.; Suart, V.; Head, E.; Clemenston, L.; Fishwick, J.; Llewellyn, C. & Aiken, J. The Second SeaWIFS HPLC Analysis Round-Robin Experiment (SeaHARRE-2) NASA/TM-2005-212785, August 2005 - Jeffrey S., Mantoura R. and Wright S. (ed.) (2005) Phytoplankton Pigments in Oceanography. Guidelines to Modern Methods, UNESCO. - Mackey, M.; Mackey, D.; Higgins, H. & Wright, S. CHEMTAX - a program for estimating class abundances from chemical markers: application to HPLC measurements of phytoplankton. Marine Ecology Progress Series, 1996, 144, 265-283

Quantitative analysis of UV-A shock and short term stress using iTRAQ, pseudo selective reaction monitoring (pSRM) and GC-MS based metabolite analysis of the cyanobacterium Nostoc punctiforme ATCC 29133.

PubMed

Wase, Nishikant; Pham, Trong Khoa; Ow, Saw Yen; Wright, Phillip C

2014-09-23

A quantitative proteomics and metabolomics analysis was performed using iTRAQ, HPLC and GC-MS in the filamentous cyanobacterium Nostoc punctiforme ATCC 29133 to understand the effect of short and long term UV-A exposure. Changes in the proteome were measured for short-term stress (4-24h) using iTRAQ. Changes in the photosynthetic pigments and intracellular metabolites were observed at exposures of up to 7days (pigments) and up to 11days (intracellular metabolites). To assess iTRAQ measurement quality, pseudo selected reaction monitoring (pSRM) was used, with this confirming underestimation of protein abundance levels by iTRAQ. Our results suggest that short term UV-A radiation lowers the abundance of PS-I and PS-II proteins. We also observed an increase in abundance of intracellular redox homeostasis proteins and plastocyanin. Additionally, we observed statistically significant changes in scytonemin, Chlorophyll A, astaxanthin, zeaxanthin, and β-carotene. Assessment of intracellular metabolites showed significant changes in several, suggesting their potential role in the Nostoc's stress mitigation strategy. Cyanobacteria under UV-A radiation have reduced growth due to intensive damage to essential functions, but the organism shows a defense response by remodeling bioenergetics pathway, induction of the UV protection compound scytonemin and increased levels of proline and tyrosine as a mitigation response. The effect of UV-A radiation on the proteome and intracellular metabolites of N. punctiforme ATCC 29133 including photosynthetic pigments has been described. We also verify the expression of 13 iTRAQ quantified protein using LC-pSRM. Overall we observed that UV-A radiation has a drastic effect on the photosynthetic machinery, photosynthetic pigments and intracellular amino acids. As a mitigation strategy against UV-A radiation, proline, glycine, and tyrosine were accumulated. Copyright © 2014. Published by Elsevier B.V.

Aging effect on the pigment composition and color of Vitis vinifera L. Cv. Tannat wines. Contribution of the main pigment families to wine color.

PubMed

Boido, Eduardo; Alcalde-Eon, Cristina; Carrau, Francisco; Dellacassa, Eduardo; Rivas-Gonzalo, Julian C

2006-09-06

Red wines made from Vitis vinifera L. cv. Tannat grapes are known to possess high contents of tannins and intense color, features that are responsible for the originality of these wines. This work aimed to study the evolution of the pigment composition and CIELAB color parameters as Tannat wines become older, as well as to establish the contribution to wine color of the main pigment families. Tannat wines produced in Uruguay from grapes of the same vineyard in six consecutive vintages (1998-2003) and Tannat grapes of the 2003 harvest were analyzed by means of HPLC-DAD-MS and UV-vis spectrometric techniques. The correlations between the different pigment families and the CIELAB parameters revealed the importance of the variations of the percentage, found in anthocyanins and flavanol-anthocyanin acetaldehyde-mediated condensation products (decrease) and pyranoanthocyanins and direct condensation products (increase), in the modification of the color from purple-red hues to more orange-red ones. The color suffered qualitative rather than quantitative changes, that is, the hue (h*ab) increased, whereas the chroma (C*ab) and lightness (L) did not show a defined trend with time.

Quantum Yields of Soluble Particulate Material in the Ocean

DTIC Science & Technology

1998-01-01

Mycosporine - like amino acids (MAAs) were separated by reverse-phase isocratic HPLC on a... Mycosporine - like Amino Acids in Phaeocystis antarctica Karsten. Marine Biology (1999b) (Submitted) Moisan, T.A., Olaizola, M. & Mitchell, B.G. Xanthophyll...monochromator and detector. We have carried out work to evaluate the role of photoprotective pigments including mycosporine amino acids and the

Red Wine Tannin Structure-Activity Relationships during Fermentation and Maceration.

PubMed

Yacco, Ralph S; Watrelot, Aude A; Kennedy, James A

2016-02-03

The correlation between tannin structure and corresponding activity was investigated by measuring the thermodynamics of interaction between tannins isolated from commercial red wine fermentations and a polystyrene divinylbenzene HPLC column. Must and/or wine samples were collected throughout fermentation/maceration from five Napa Valley wineries. By varying winery, fruit source, maceration time, and cap management practice, it was considered that a reasonably large variation in commercially relevant tannin structure would result. Tannins were isolated from samples collected using low pressure chromatography and were then characterized by gel permeation chromatography and acid-catalyzed cleavage in the presence of excess phloroglucinol (phloroglucinolysis). Corresponding tannin activity was determined using HPLC by measuring the thermodynamics of interaction between isolated tannin and a polystyrene divinylbenzene HPLC column. This measurement approach was designed to determine the ability of tannins to hydrophobically interact with a hydrophobic surface. The results of this study indicate that tannin activity is primarily driven by molecular size. Compositionally, tannin activity was positively associated with seed tannins and negatively associated with skin and pigmented tannins. Although measured indirectly, the extent of tannin oxidation as determined by phloroglucinolysis conversion yield suggests that tannin oxidation at this stage of production reduces tannin activity. Based upon maceration time, this study indicates that observed increases in perceived astringency quality, if related to tannin chemistry, are driven by tannin molecular mass as opposed to pigmented tannin formation or oxidation. Overall, the results of this study give new insight into tannin structure-activity relationships which dominate during extraction.

Ultratraces of carotenes in tomato purées: HPLC-TLS study

NASA Astrophysics Data System (ADS)

Luterotti, S.; Marković, K.; Franko, M.; Bicanic, D.; Vahčić, N.; Doka, O.

2003-01-01

The present study was designed to provide information about (i) the profile of carotene pigments and (ii) trace quantities of lycopene and β-carotene left in tomato purées. The ultrasensitive method comprising HPLC and thermal lens spectrometric (TLS) detection enabled us to detect as low as 0.3 and 1.1 ng ml-1 lycopene and β-carotene in purée extracts, respectively. Total concentration of β-carotene and lycopene (varying from 3 to 170 ng g-1) in the examined tomato purées may serve as an indicator of the carotene-specific antioxidative capacity of these products. Although conventional spectrophotometry can be used to rapidly assess the quality of products derived from tomatoes, a highly sensitive and selective method such as HPLC-TLS is needed for reliable analyses of samples such as, for example, those subjected to inappropriate storage and/or handling.

Separation of amaranthine-type betacyanins by ion-pair high-speed countercurrent chromatography.

PubMed

Jerz, Gerold; Gebers, Nadine; Szot, Dominika; Szaleniec, Maciej; Winterhalter, Peter; Wybraniec, Slawomir

2014-05-30

Betacyanins, red-violet plant pigments, were fractionated by ion-pair high-speed countercurrent chromatography (IP-HSCCC) from leaves extract of Iresine lindenii Van Houtte, an ornamental plant of the family Amaranthaceae. An HSCCC solvent system consisting of TBME-1-BuOH-ACN-H2O (1:3:1:5, v/v/v/v) was applied using ion-pair forming heptafluorobutyric acid (HFBA). Significantly different elution profiles of betacyanin diastereomeric pairs (derivatives based on betanidin and isobetanidin) observed in the HSCCC in comparison to HPLC systems indicate a complementarity of both techniques' fractionation capabilities. The numerous diastereomeric pairs can be selectively separated from each other using the HSCCC system simplifying the pigment purification process. Apart from the three well known highly abundant pigments (amaranthine, betanin and iresinin I) together with their isoforms, three new acylated (feruloylated and sinapoylated) betacyanins as well as known pigment hylocerenin (previously isolated from cacti fruits) were characterized in the plant for the first time and they are new for the whole Amaranthaceae family. Copyright © 2014 Elsevier B.V. All rights reserved.

Microencapsulation of betalains obtained from cactus fruit (Opuntia ficus-indica) by spray drying using cactus cladode mucilage and maltodextrin as encapsulating agents.

PubMed

Otálora, María Carolina; Carriazo, José Gregorio; Iturriaga, Laura; Nazareno, Mónica Azucena; Osorio, Coralia

2015-11-15

The microencapsulation of betalains from cactus fruit by spray drying was evaluated as a stabilization strategy for these pigments. The betalains used as active agent were extracted from purple fruits of Opuntia ficus-indica (BE) and encapsulated with maltodextrin and cladode mucilage MD-CM and only with MD. The microcapsulates were characterized by scanning electron microscopy (SEM), thermal analysis (TGA-DSC), tristimulus colorimetry, as well as, their humidity, water activity and dietary fiber content were also determined. The active agent content was measured by UV-Vis spectrophotometry and its composition confirmed by HPLC-ESIMS. A pigment storage stability test was performed at 18 °C and different relative humidities. The addition of CM in the formulation increased the encapsulation efficiency, diminished the moisture content, and allowed to obtain more uniform size and spherical particles, with high dietary fiber content. These microencapsulates are promising functional additive to be used as natural colorant in the food industry. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effect of metal stress on photosynthetic pigments in the Cu-hyperaccumulating lichens Cladonia humilis and Stereocaulon japonicum growing in Cu-polluted sites in Japan.

PubMed

Nakajima, Hiromitsu; Yamamoto, Yoshikazu; Yoshitani, Azusa; Itoh, Kiminori

2013-11-01

To understand the ecology and physiology of metal-accumulating lichens growing in Cu-polluted sites, we investigated lichens near temple and shrine buildings with Cu roofs in Japan and found that Stereocaulon japonicum Th. Fr. and Cladonia humilis (With.) J. R. Laundon grow in Cu-polluted sites. Metal concentrations in the lichen samples collected at some of these sites were determined by inductively coupled plasma mass spectroscopy (ICP-MS). UV-vis absorption spectra of pigments extracted from the lichen samples were measured, and the pigment concentrations were estimated from the spectral data using equations from the literature. Secondary metabolites extracted from the lichen samples were analyzed by high-performance liquid chromatography (HPLC) with a photodiode array detector. We found that S. japonicum and C. humilis are Cu-hyperaccumulating lichens. Differences in pigment concentrations and their absorption spectra were observed between the Cu-polluted and control samples of the 2 lichens. However, no correlation was found between Cu and pigment concentrations. We observed a positive correlation between Al and Fe concentrations and unexpectedly found high negative correlations between Al and pigment concentrations. This suggests that Al stress reduces pigment concentrations. The concentrations of secondary metabolites in C. humilis growing in the Cu-polluted sites agreed with those in C. humilis growing in the control sites. This indicates that the metabolite concentrations are independent of Cu stress. Copyright © 2013 Elsevier Inc. All rights reserved.

Separation and Identification of Anthocyanins Extracted from Blueberry Wine Lees and Pigment Binding Properties toward β-Glucosidase.

PubMed

Wu, Qian; Zhang, Yang; Tang, Hu; Chen, Yashu; Xie, Bijun; Wang, Chao; Sun, Zhida

2017-01-11

Anthocyanins were isolated from blueberry wine lees using Sephadex LH-20 column chromatography and semipreparative high-performance liquid chromatography (semipreparative HPLC) and then identified by HPLC-DAD-ESI-MS/MS. Our results show that malvidin-3-hexose (Mv-3-hex) and malvidin-3-(6'acetyl)-hexose (Mv-3-ace-hex) are the major components in the anthocyanin extracts of blueberry wine lees (>90%). The binding characteristics of Mv-3-hex and Mv-3-ace-hex with β-glucosidase were investigated by fluorescence spectroscopy, circular dichroism (CD) spectroscopy, and molecular docking. Spectroscopic analysis revealed that β-glucosidase fluorescence quenched by Mv-3-hex and Mv-3-ace-hex follows a static mode. Binding of Mv-3-hex and Mv-3-ace-hex to β-glucosidase mainly depends on electrostatic force. The result from CD spectra shows that adaptive structure rearrangement and increase of β-sheet structure occur only in the presence of Mv-3-ace-hex. A molecular docking study suggests that Mv-3-ace-hex has stronger binding with β-glucosidase than Mv-3-hex.

Growth-inhibitory effects of pigmented rice bran extracts and three red bran fractions against human cancer cells: Relationships to composition and antioxidative activities

USDA-ARS?s Scientific Manuscript database

We determined the phenolic, anthocyanin, and proanthocyanidin content of three brown, purple, and red rice brans isolated from different rice varieties using HPLC-PDA with the aid of 27 standards of known structure and by matching unknown peaks to a spectral library of known compounds. DPPH and ORA...

Carotenoids of two freshwater amphipod species (Gammarus pulex and G. roeseli) and their common acanthocephalan parasite Polymorphus minutus.

PubMed

Gaillard, Maria; Juillet, Cédrik; Cézilly, Frank; Perrot-Minnot, Marie-Jeanne

2004-09-01

Carotenoid compositions of two freshwater Gammarus species (Crustacea: Amphipoda) and of their common acanthocephalan parasite Polymorphus minutus were characterized. The effect of carotenoid uptake by the parasite was addressed by comparing the carotenoid content of uninfected and infected female hosts. Using high-pressure liquid chromatography (HPLC), co-chromatography of reference pigments and electron ionization mass spectrometry of collected HPLC fractions (EI-MS), several xanthophylls and non-polar compounds were identified. Seven kinds of carotenoids, mainly xanthophylls, were identified in gammarids. Astaxanthin was predominant, amounting to 40 wt.% of total carotenoid in both uninfected G. pulex and G. roeseli. By contrast, we found only non-polar compounds with a predominance of esterified forms of astaxanthin in P. minutus larvae. No significant effect of infection on carotenoid content was evidenced in G. pulex and G. roeseli females. Our study highlights the use of a Matrix Solid Phase Dispersion as an efficient extraction method of both xanthophylls and non-polar pigments in small samples, including lipid-rich ones as P. minutus parasite. We discuss on the presumptive pathway leading to the formation of free astaxanthin in gammarids via hydroxy compounds, and on the accumulation of esters of astaxanthin in parasites.

Spatial distribution of Chloroflexus-like bacteria in the hypersaline artificial microbial mat

NASA Astrophysics Data System (ADS)

Bachar, A.; Polerecky, L.; Vamvakopoulos, K.; de Beer, D.; Jonkers, H. M.

An artificial microbial mat grown in a mesocosm originated from the Hypersaline Lake of La Salada de Chiprana NE Spain was examined with respect to its organism s spatial distribution via high resolution methods A special attention was given to the elucidative Chloroflexus -like bacteria on which spatial distribution data is not available We have characterized this thick 1cm and developed mat for photopigments HPLC and obtained the general pigment distribution pattern Furthermore fiberoptic and photosynthetic microsensor measurements gave inner light attenuations and flux rates of oxygen within the different layers respectively Using fluorescence and spectral imaging we were able to detect characteristic pigmentation in the different layers FISH probes targeting Chloroflexus -like bacteria confirmed the visualization techniques and showed a single hybridized layer below the cyanobacterial layer as did the HPLC fiberoptic microsensor and fluorescence imaging We conclude that Chloroflexus -like bacteria are located below the cyanobacterial layer and above the purple sulfur bacteria and for the firs time we are able to show it by different independent state of the art techniques These approaches can be important for rapid community investigations within a millimeter scale microniches

Sea-ice eukaryotes of the Gulf of Finland, Baltic Sea, and evidence for herbivory on weakly shade-adapted ice algae.

PubMed

Majaneva, Markus; Blomster, Jaanika; Müller, Susann; Autio, Riitta; Majaneva, Sanna; Hyytiäinen, Kirsi; Nagai, Satoshi; Rintala, Janne-Markus

2017-02-01

To determine community composition and physiological status of early spring sea-ice organisms, we collected sea-ice, slush and under-ice water samples from the Baltic Sea. We combined light microscopy, HPLC pigment analysis and pyrosequencing, and related the biomass and physiological status of sea-ice algae with the protistan community composition in a new way in the area. In terms of biomass, centric diatoms including a distinct Melosira arctica bloom in the upper intermediate section of the fast ice, dinoflagellates, euglenoids and the cyanobacterium Aphanizomenon sp. predominated in the sea-ice sections and unidentified flagellates in the slush. Based on pigment analyses, the ice-algal communities showed no adjusted photosynthetic pigment pools throughout the sea ice, and the bottom-ice communities were not shade-adapted. The sea ice included more characteristic phototrophic taxa (49%) than did slush (18%) and under-ice water (37%). Cercozoans and ciliates were the richest taxon groups, and the differences among the communities arose mainly from the various phagotrophic protistan taxa inhabiting the communities. The presence of pheophytin a coincided with an elevated ciliate biomass and read abundance in the drift ice and with a high Eurytemora affinis read abundance in the pack ice, indicating that ciliates and Eurytemora affinis were grazing on algae. Copyright © 2016 Elsevier GmbH. All rights reserved.

Photosynthetic pigment fingerprints as indicators of phytoplankton biomass and development in different water masses of the Southern Ocean during austral spring

NASA Astrophysics Data System (ADS)

Peeken, Ilka

The development of phytoplankton biomass and composition was investigated on three occasions along a longitudinal transect (6°W) between 60°S and 47°S from October 13 to November 21, 1992 by measurement of photosynthetic pigments with high performance liquid chromatography (HPLC). Measured accessory pigment concentrations were multiplied by conversion factors to derive the proportions of phytoplankton groups contributing to the biomass indicator chlorophyll a. Phytoplankton blooms developed in the Polar Frontal region (PFr) and were dominated (80%) by diatoms. Other groups contributing to the phytoplankton included prymnesiophytes, green algae, autotrophic dinoflagellates, cryptophytes, pelagophytes and micromonadophytes, and their distributions varied with time. In contrast, phytoplankton biomass remained low in the southern Antarctic Circumpolar Current (ACC) and was dominated by flagellates, particularly green algae and prymnesiophytes. Green algae contributed more to total biomass than in previous investigations, partly attributed to "Chlorella-like" type organisms rather than prasinophytes. Cryptophytes decreased during the investigation, possibly due to salp grazing. No bloom was observed at the retreating ice-edge, presumably due to strong wind mixing. Only a slight increase in phytoplankton biomass, composed primarily of diatoms, was found at the ACC-Weddell Gyre front. Cluster analysis revealed that different phytoplankton communities characterised the different water masses of the PFr and southern ACC; the history of different water masses in the PFr could be reconstructed on this basis.

Optically-derived estimates of phytoplankton size class and taxonomic group biomass in the Eastern Subarctic Pacific Ocean

NASA Astrophysics Data System (ADS)

Zeng, Chen; Rosengard, Sarah Z.; Burt, William; Peña, M. Angelica; Nemcek, Nina; Zeng, Tao; Arrigo, Kevin R.; Tortell, Philippe D.

2018-06-01

We evaluate several algorithms for the estimation of phytoplankton size class (PSC) and functional type (PFT) biomass from ship-based optical measurements in the Subarctic Northeast Pacific Ocean. Using underway measurements of particulate absorption and backscatter in surface waters, we derived estimates of PSC/PFT based on chlorophyll-a concentrations (Chl-a), particulate absorption spectra and the wavelength dependence of particulate backscatter. Optically-derived [Chl-a] and phytoplankton absorption measurements were validated against discrete calibration samples, while the derived PSC/PFT estimates were validated using size-fractionated Chl-a measurements and HPLC analysis of diagnostic photosynthetic pigments (DPA). Our results showflo that PSC/PFT algorithms based on [Chl-a] and particulate absorption spectra performed significantly better than the backscatter slope approach. These two more successful algorithms yielded estimates of phytoplankton size classes that agreed well with HPLC-derived DPA estimates (RMSE = 12.9%, and 16.6%, respectively) across a range of hydrographic and productivity regimes. Moreover, the [Chl-a] algorithm produced PSC estimates that agreed well with size-fractionated [Chl-a] measurements, and estimates of the biomass of specific phytoplankton groups that were consistent with values derived from HPLC. Based on these results, we suggest that simple [Chl-a] measurements should be more fully exploited to improve the classification of phytoplankton assemblages in the Northeast Pacific Ocean.

Study and Characterization of an Ancient European Flint White Maize Rich in Anthocyanins: Millo Corvo from Galicia

PubMed Central

Lago, Chiara; Landoni, Michela; Cassani, Elena; Cantaluppi, Enrico; Doria, Enrico; Nielsen, Erik; Giorgi, Annamaria; Pilu, Roberto

2015-01-01

In the second half of the last century, the American dent hybrids began to be widely grown, leading to the disappearance or marginalization of the less productive traditional varieties. Nowadays the characterization of traditional landraces can help breeders to discover precious alleles that could be useful for modern genetic improvement and allow a correct conservation of these open pollinated varieties (opvs). In this work we characterized the ancient coloured cultivar “Millo Corvo” typical of the Spanish region of Galicia. We showed that this cultivar accumulates high amounts of anthocyanins (83.4 mg/100g flour), and by TLC (Thin Layer Chromatography) and HPLC (High Pressure Liquid Chromatography) analysis, we demonstrated that they mainly consisted of cyanidin. Mapping and sequencing data demonstrate that anthocyanin pigmentation is due to the presence of the red color1 gene(r1), a transcription factor driving the accumulation of this pigment in the aleurone layer. Further chemical analysis showed that the kernels are lacking in carotenoids, as confirmed by genetic study. Finally a DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging ability test showed that Millo Corvo, even though lacking carotenoids, has a high antioxidant ability, and could be considered as a functional food due to the presence of anthocyanins. PMID:25961304

Overexpression of the rice carotenoid cleavage dioxygenase 1 gene in Golden Rice endosperm suggests apocarotenoids as substrates in planta.

PubMed

Ilg, Andrea; Yu, Qiuju; Schaub, Patrick; Beyer, Peter; Al-Babili, Salim

2010-08-01

Carotenoids are converted by carotenoid cleavage dioxygenases that catalyze oxidative cleavage reactions leading to apocarotenoids. However, apocarotenoids can also be further truncated by some members of this enzyme family. The plant carotenoid cleavage dioxygenase 1 (CCD1) subfamily is known to degrade both carotenoids and apocarotenoids in vitro, leading to different volatile compounds. In this study, we investigated the impact of the rice CCD1 (OsCCD1) on the pigmentation of Golden Rice 2 (GR2), a genetically modified rice variety accumulating carotenoids in the endosperm. For this purpose, the corresponding cDNA was introduced into the rice genome under the control of an endosperm-specific promoter in sense and anti-sense orientations. Despite high expression levels of OsCCD1 in sense plants, pigment analysis revealed carotenoid levels and patterns comparable to those of GR2, pleading against carotenoids as substrates in rice endosperm. In support, similar carotenoid contents were determined in anti-sense plants. To check whether OsCCD1 overexpressed in GR2 endosperm is active, in vitro assays were performed with apocarotenoid substrates. HPLC analysis confirmed the cleavage activity of introduced OsCCD1. Our data indicate that apocarotenoids rather than carotenoids are the substrates of OsCCD1 in planta.

Sources of Variability in Chlorophyll Analysis by Fluorometry and by High Performance Liquid Chromatography. Chapter 22

NASA Technical Reports Server (NTRS)

VanHeukelem, Laurie; Thomas, Crystal S.; Glibert, Patricia M.

2001-01-01

The need for accurate determination of chlorophyll a (chl a) is of interest for numerous reasons. From the need for ground-truth data for remote sensing to pigment detection for laboratory experimentation, it is essential to know the accuracy of the analyses and the factors potentially contributing to variability and error. Numerous methods and instrument techniques are currently employed in the analyses of chl a. These methods range from spectrophotometric quantification, to fluorometric analysis and determination by high performance liquid chromatography. Even within the application of HPLC techniques, methods vary. Here we provide the results of a comparison among methods and provide some guidance for improving the accuracy of these analyses. These results are based on a round-robin conducted among numerous investigators, including several in the Sensor Intercomparison and Merger for Biological and Interdisciplinary Oceanic Studies (SIMBIOS) and HyCODE Programs. Our purpose here is not to present the full results of the laboratory intercalibration; those results will be presented elsewhere. Rather, here we highlight some of the major factors that may contribute to the variability observed. Specifically, we aim to assess the comparability of chl a analyses performed by fluorometry and HPLC, and we identify several factors in the analyses which may contribute disproportionately to this variability.

Temporal variation in photosynthetic pigments and UV-absorbing compounds in shallow populations of two Hawaiian reef corals

USGS Publications Warehouse

Kuffner, I.B.

2005-01-01

As we seek to understand the physiological mechanisms of coral bleaching, it is important to understand the background temporal variation in photosynthetic pigments and photoprotective compounds that corals exhibit. In this study, reef flat populations of two hermatypic coral species, Montipora capitata (Dana, 1846) and Porites compressa Dana, 1846, were sampled monthly in Kane'ohe Bay, Hawai'i, from January 1998 to March 1999. Surface ultraviolet radiation (UVR) was measured continually during this time period at the same location. High-performance liquid chromatography (HPLC) analysis of photosynthetic pigments and mycosporine-like amino acids (MAAs) revealed temporal changes in concentrations and proportions of these compounds in tissues of both species of coral. Chlorophyll a (chl a), chlorophyll c2 (chl c2), peridinin, and diadinoxanthin concentrations changed on a skeletal weight (M. capitata) or surface area (P. compressa) basis, significantly correlating with seasonal changes in solar input (number of days from the winter solstice). In P. compressa, diadinoxanthin increased in proportion to the total pigment pool during summer months, suggesting an up-regulation of a xanthophyll cycle. In M. capitata, the ratio of chl a: chl c2 decreased during winter months, suggesting photoacclimation to lower light levels. It is surprising that there was not a clear seasonal pattern in total MAA concentration for either species, with the exception of shinorine in P. compressa. The relative stability of MAA concentrations over the course of the year despite a pronounced seasonal trend in UVR suggests either that MAAs are not performing a photoprotective role in these species or that concentrations are kept at a threshold level in the presence of a dynamic light environment. ?? 2005 by University of Hawai'i Press All rights reserved.

Characterization of pigments from different high speed countercurrent chromatography wine fractions.

PubMed

Salas, Erika; Dueñas, Montserrat; Schwarz, Michael; Winterhalter, Peter; Cheynier, Véronique; Fulcrand, Hélène

2005-06-01

A red wine, made from Cabernet Sauvignon (60%) and Tannat (40%) cultivars, was fractionated by high speed countercurrent chromatography (HSCCC). The biphasic solvent system consisting of tert-butyl methyl ether/n-butanol/acetonitrile/water (2/2/1/5, acidified with 0.1% trifluoroacetic acid) was chosen for its demonstrated efficiency in separating anthocyanins. The different native and derived anthocyanins were identified on the basis of their UV-visible spectra, their elution time on reversed-phase high-performance liquid chromatography (HPLC), and their mass spectra, before and after thiolysis. The HSCCC method allowed the separation of different families of anthocyanin-derived pigments that were eluted in different fractions according to their structures. The hydrosoluble fraction was almost devoid of native anthocyanins. Further characterization (glucose quantification, UV-visible absorbance measurements) indicated that it contained flavanol and anthocyanin copolymers in which parts of the anthocyanin units were in colorless forms. Pigments in the hydrosoluble fraction showed increased resistance to sulfite bleaching and to the nucleophilic attack of water.

Identification of an indigo precursor from leaves of Isatis tinctoria (Woad).

PubMed

Maugard, T; Enaud, E; Choisy, P; Legoy, M D

2001-11-01

Indole is presumably a product of indole-3-glycerol phosphate catabolism in Isatis tinctoria. It is oxidized into indoxyl and stored in young leaves as indigo precursor. Further oxidation and dimerization of indoxyl produces indigoid pigments. In this work, we describe an HPLC method dedicated to the identification and quantification of indigoid pigments (indigo, indirubin, isoindigo and isoindirubin) and indigo precursors produced in I. tinctoria (Woad). This work, carried out with two cultivars of I. tinctoria, has confirmed that the quantity of indigo precursors is dependent on the species and the harvest period. In addition we have shown for the first time that young leaves of I. tinctoria, harvested in June contained a new indigo precursor in addition to isatan B (indoxyl-5-ketogluconate) and indican (indoxyl-beta-D-glucoside). We suggest the name "isatan C" for this new indigo precursor in I. tinctoria. Its chemical characteristics point to an dioxindole ester with PM of 395. We have shown that isatan C reacts with isatan B increasing the red pigment production.

Chlorophyll and carotenoid pigments of prochloron (prochlorophyta)

NASA Technical Reports Server (NTRS)

Paerl, H. W.; Lewin, R. A.; Cheng, L.

1983-01-01

High-performance liquid chromatography (HPLC) with a gradient-elution technique was utilized to separate and quantify chlorophylls a and b as well as major carotenoid pigments present in freeze-dried preprations of prochloron-didemnid associations and in Prochloron cells separated from host colonies. Results confirm earlier spectrophotometric evidence for both chlorophylls a and b in this prokaryote. Chlorophyll a:b ratios range from 4.14 to 19.71; generally good agreement was found between ratios determined in isolated cell preprations and in symbiotic colonies (in hospite). These values are 1.5 to 5-fold higher than ratios determined in a variety of eukaryotic green plants. The carotenoids in Prochloron are quantitatively and qualitatively similar to those found in various freshwater and marine blue-green algae (cyanopbytes) from high-light environments. However, Prochloron differs from cyanophytes by the absence of myxoxanthophyll and related glycosidic carotenoids. It pigment characteristics are considered sufficiently different from those of cyanophytes to justify its assignment to a separate algal division.

Preparation and Antioxidant Activity of Ethyl-Linked Anthocyanin-Flavanol Pigments from Model Wine Solutions.

PubMed

Li, Lingxi; Zhang, Minna; Zhang, Shuting; Cui, Yan; Sun, Baoshan

2018-05-03

Anthocyanin-flavanol pigments, formed during red wine fermentation and storage by condensation reactions between anthocyanins and flavanols (monomers, oligomers, and polymers), are one of the major groups of polyphenols in aged red wine. However, knowledge of their biological activities is lacking. This is probably due to the structural diversity and complexity of these molecules, which makes the large-scale separation and isolation of the individual compounds very difficult, thus restricting their further study. In this study, anthocyanins (i.e., malvidin-3-glucoside, cyanidin-3-glucoside, and peonidin-3-glucoside) and (⁻)-epicatechin were first isolated at a preparative scale by high-speed counter-current chromatography. The condensation reaction between each of the isolated anthocyanins and (⁻)-epicatechin, mediated by acetaldehyde, was conducted in model wine solutions to obtain ethyl-linked anthocyanin-flavanol pigments. The effects of pH, molar ratio, and temperature on the reaction rate were investigated, and the reaction conditions of pH 1.7, molar ratio 1:6:10 (anthocyanin/(⁻)-epicatechin/acetaldehyde), and reaction temperature of 35 °C were identified as optimal for conversion of anthocyanins to ethyl-linked anthocyanin-flavanol pigments. Six ethyl-linked anthocyanin-flavanol pigments were isolated in larger quantities and collected under optimal reaction conditions, and their chemical structures were identified by HPLC-QTOF-MS and ECD analyses. Furthermore, DPPH, ABTS, and FRAP assays indicate that ethyl-linked anthocyanin-flavanol pigments show stronger antioxidant activities than their precursor anthocyanins.

Improved HPLC method for the determination of curcumin, demethoxycurcumin, and bisdemethoxycurcumin.

PubMed

Jayaprakasha, Guddadarangavvanahally K; Jagan Mohan Rao, Lingamullu; Sakariah, Kunnumpurath K

2002-06-19

Commercially available curcumin, a bright orange-yellow color pigment of turmeric, consists of a mixture of three curcuminoids, namely, curcumin, demethoxycurcumin, and bisdemethoxycurcumin. These were isolated by column chromatography and identified by spectroscopic studies. The purity of the curcuminoids was analyzed by an improved HPLC method. HPLC separation was performed on a C(18) column using three solvents, methanol, 2% AcOH, and acetonitrile, with detection at 425 nm. Four different commercially available varieties of turmeric, namely, Salem, Erode, Balasore, and local market samples, were analyzed to detect the percentage of these three curcuminoids. The percentages of curcumin, demethoxycurcumin, and bisdemethoxycurcumin as estimated using their calibration curves were found to be 1.06 +/- 0.061 to 5.65 +/- 0.040, 0.83 +/- 0.047 to 3.36 +/- 0.040, and 0.42 +/- 0.036 to 2.16 +/- 0.06, respectively, in four different samples. The total percentages of curcuminoids are 2.34 +/- 0.171 to 9.18 +/- 0.232%.

Hyper-spectral imaging: A promising tool for quantitative pigment analysis of varved lake sediments

NASA Astrophysics Data System (ADS)

Butz, Christoph; Grosjean, Martin; Tylmann, Wojciech

2015-04-01

Varved lake sediments are good archives for past environmental and climate conditions from annual to multi-millennial scales. Among other proxies, concentrations of sedimentary photopigments have been used for temperature reconstructions. However, obtaining well calibrated annually resolved records from sediments still remains challenging. Most laboratory methods used to analyse lake sediments require physical subsampling and are destructive in the process. Hence, temporal resolution and number of data are limited by the amount of material available in the core. Furthermore, for very low sediment accumulation rates annual subsampling is often very difficult or even impossible. To address these problems we explore hyper-spectral imaging as a non-destructive method to analyse lake sediments based on their reflectance spectra in the visible and near infrared spectrum. In contrast to other scanning methods like X-ray fluorescence, VIS/NIR reflectance spectrometry distinguishes between biogeochemical substances rather than single elements. Among others Rein (2003) has shown that VIS-RS can be used to detect relative concentrations of sedimentary photopigments (e.g. chlorins, carotenoids) and clay minerals. In this study hyper-spectral imaging is used to infer ecological proxy data from reflectance spectra of varved lake sediments. Hyper-spectral imaging permits the measurement of an entire sediment core in a single run at high spatial (30x30µm/pixel) and spectral resolutions (~2.8nm) within the visual to near infrared spectrum (400-1000nm). This allows the analysis of data time series and spatial mapping of sedimentary substances (e.g. chlorophylls/bacterio-chlorophylls and diagenetic products) at sub-varve scales. The method is demonstrated on two varved lake sediments from northern Poland showing the distributions of relative concentrations of two types of sedimentary pigments (Chlorophyll-a + derivatives and Bacterio-pheophytin-a) within individual varve years. The relative concentrations from the spectral data set have then been calibrated with absolute concentrations derived by High-Performance-Liquid-Chromatography (HPLC). This results in very high-resolution data sets of absolute sedimentary pigment concentrations suitable for the analysis of seasonal pigment variations.

An Investigation Into HPLC Data Quality Problems

NASA Technical Reports Server (NTRS)

Hooker, Stanford B.; VanHeukelem, Laurie

2011-01-01

This report summarizes the analyses and results produced by a five-member investigative team of Government, university, and industry experts, established by NASA HQ. The team examined data quality problems associated with high performance liquid chromatography (HPLC) analyses of pigment concentrations in seawater samples produced by the San Diego State University (SDSU) Center for Hydro-Optics and Remote Sensing (CHORS). This report shows CHORS did not validate the methods used before placing them into service to analyze field samples for NASA principal investigators (PIs), even though the HPLC literature contained easily accessible method validation procedures, and the importance of implementing them, more than a decade ago. In addition, there were so many sources of significant variance in the CHORS methodologies, that the HPLC system rarely operated within performance criteria capable of producing the requisite data quality. It is the recommendation of the investigative team to a) not correct the data, b) make all the data that was temporarily sequestered available for scientific use, and c) label the affected data with an appropriate warning, e.g., "These data are not validated and should not be used as the sole basis for a scientific result, conclusion, or hypothesis--independent corroborating evidence is required."

Pink berry grape (Vitis vinifera L.) characterization: Reflectance spectroscopy, HPLC and molecular markers.

PubMed

Rustioni, Laura; De Lorenzis, Gabriella; Hârţa, Monica; Failla, Osvaldo

2016-01-01

Color has a fundamental role for the qualitative evaluation and cultivar characterization of fruits. In grape, a normally functional pigment biosynthesis leads to the accumulation of a high quantity of anthocyanins. In this work, 28 Vitis vinifera L. cultivars accumulating low anthocyanins in berries were studied to characterize the biosynthetic dysfunctions in both a phenotypic and genotypic point of view. Reflectance spectroscopy, HPLC profiles and molecular markers related to VvMybA1 and VvMybA2 genes allowed a detailed description of the pigment-related characteristics of these cultivars. Data were consistent concerning the heterozygosity of the non-functional allele in both investigated genes, resulting in a low colored phenotype as described by reflectance. However, the variability in berry colour among our samples was not fully explained by MybA locus, probably due to specific interferences among the biosynthetic pathways, as suggested by the anthocyanin profile variations detected among our samples. The results presented in this work confirmed the importance of the genetic background: grapes accumulating high levels of cyanidin-3-O-glucosides (di-substituted anthocyanin) are generally originated by white cultivar retro-mutations and they seem to preserve the anomalies in the flavonoid hydroxylases enzymes which negatively affect the synthesis of tri-substituted anthocyanins. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

Using CHEMTAX to evaluate seasonal and interannual dynamics of the phytoplankton community off the South-west coast of Portugal

NASA Astrophysics Data System (ADS)

Goela, P. C.; Danchenko, S.; Icely, J. D.; Lubian, L. M.; Cristina, S.; Newton, A.

2014-12-01

CHEMTAX was used to assess the relative contribution of the main phytoplankton classes to the total concentration of Chlorophyll a (Chl a) from the waters off SW coast of Portugal. Sampling campaigns were carried out during all seasons from 2008 to 2012, at three stations located 2, 10 and 18 km from the coast. Samples were taken from the surface, mid-Secchi and Secchi depth, for the determination of Chl a and other phytoplanktonic pigments by HPLC. Supporting data were also obtained including dissolved inorganic nutrients, salinity, transparency, temperature and upwelling indices. The CHEMTAX results were also related to microscopy counts and also spectral analysis of absorption of other samples from the same sampling campaigns. The pigment results showed that diatoms dominated from early spring to summer, coinciding with upwelling conditions, while cryptophytes, prymnesiophytes and prasinophytes dominated in autumn and winter, coinciding with seasonal stratification. Although the contribution of cyanobacteria to total Chl a was generally low, there were occasional sampling campaigns where it was exceptionally high, but these appeared not to be related to upwelling. Dinoflagellates and chrysophytes were minority groups although the pigment marker peridinin that was used to distinguish dinoflagellates was not adequate for distinguishing all the members of this group. CHEMTAX was particularly useful for discriminating between the smaller (0-20 μm) classes of the microplankton that could not be easily identified by microscopy.

Physico-Chemical Evaluation of Rationally Designed Melanins as Novel Nature-Inspired Radioprotectors

PubMed Central

Schweitzer, Andrew D.; Howell, Robertha C.; Jiang, Zewei; Bryan, Ruth A.; Gerfen, Gary; Chen, Chin-Cheng; Mah, Dennis; Cahill, Sean

2009-01-01

Background Melanin, a high-molecular weight pigment that is ubiquitous in nature, protects melanized microorganisms against high doses of ionizing radiation. However, the physics of melanin interaction with ionizing radiation is unknown. Methodology/Principal Findings We rationally designed melanins from either 5-S-cysteinyl-DOPA, L-cysteine/L-DOPA, or L-DOPA with diverse structures as shown by elemental analysis and HPLC. Sulfur-containing melanins had higher predicted attenuation coefficients than non-sulfur-containing melanins. All synthetic melanins displayed strong electron paramagnetic resonance (2.14·1018, 7.09·1018, and 9.05·1017 spins/g, respectively), with sulfur-containing melanins demonstrating more complex spectra and higher numbers of stable free radicals. There was no change in the quality or quantity of the stable free radicals after high-dose (30,000 cGy), high-energy (137Cs, 661.6 keV) irradiation, indicating a high degree of radical stability as well as a robust resistance to the ionizing effects of gamma irradiation. The rationally designed melanins protected mammalian cells against ionizing radiation of different energies. Conclusions/Significance We propose that due to melanin's numerous aromatic oligomers containing multiple π-electron system, a generated Compton recoil electron gradually loses energy while passing through the pigment, until its energy is sufficiently low that it can be trapped by stable free radicals present in the pigment. Controlled dissipation of high-energy recoil electrons by melanin prevents secondary ionizations and the generation of damaging free radical species. PMID:19789711

Molecular diversity, metabolic transformation, and evolution of carotenoid feather pigments in cotingas (Aves: Cotingidae).

PubMed

Prum, Richard O; LaFountain, Amy M; Berro, Julien; Stoddard, Mary Caswell; Frank, Harry A

2012-12-01

Carotenoid pigments were extracted from 29 feather patches from 25 species of cotingas (Cotingidae) representing all lineages of the family with carotenoid plumage coloration. Using high-performance liquid chromatography (HPLC), mass spectrometry, chemical analysis, and ¹H-NMR, 16 different carotenoid molecules were documented in the plumages of the cotinga family. These included common dietary xanthophylls (lutein and zeaxanthin), canary xanthophylls A and B, four well known and broadly distributed avian ketocarotenoids (canthaxanthin, astaxanthin, α-doradexanthin, and adonixanthin), rhodoxanthin, and seven 4-methoxy-ketocarotenoids. Methoxy-ketocarotenoids were found in 12 species within seven cotinga genera, including a new, previously undescribed molecule isolated from the Andean Cock-of-the-Rock Rupicola peruviana, 3'-hydroxy-3-methoxy-β,β-carotene-4-one, which we name rupicolin. The diversity of cotinga plumage carotenoid pigments is hypothesized to be derived via four metabolic pathways from lutein, zeaxanthin, β-cryptoxanthin, and β-carotene. All metabolic transformations within the four pathways can be described by six or seven different enzymatic reactions. Three of these reactions are shared among three precursor pathways and are responsible for eight different metabolically derived carotenoid molecules. The function of cotinga plumage carotenoid diversity was analyzed with reflectance spectrophotometry of plumage patches and a tetrahedral model of avian color visual perception. The evolutionary history of the origin of this diversity is analyzed phylogenetically. The color space analyses document that the evolutionarily derived metabolic modifications of dietary xanthophylls have resulted in the creation of distinctive orange-red and purple visual colors.

Physico-chemical evaluation of rationally designed melanins as novel nature-inspired radioprotectors.

PubMed

Schweitzer, Andrew D; Howell, Robertha C; Jiang, Zewei; Bryan, Ruth A; Gerfen, Gary; Chen, Chin-Cheng; Mah, Dennis; Cahill, Sean; Casadevall, Arturo; Dadachova, Ekaterina

2009-09-30

Melanin, a high-molecular weight pigment that is ubiquitous in nature, protects melanized microorganisms against high doses of ionizing radiation. However, the physics of melanin interaction with ionizing radiation is unknown. We rationally designed melanins from either 5-S-cysteinyl-DOPA, L-cysteine/L-DOPA, or L-DOPA with diverse structures as shown by elemental analysis and HPLC. Sulfur-containing melanins had higher predicted attenuation coefficients than non-sulfur-containing melanins. All synthetic melanins displayed strong electron paramagnetic resonance (2.14.10(18), 7.09.10(18), and 9.05.10(17) spins/g, respectively), with sulfur-containing melanins demonstrating more complex spectra and higher numbers of stable free radicals. There was no change in the quality or quantity of the stable free radicals after high-dose (30,000 cGy), high-energy ((137)Cs, 661.6 keV) irradiation, indicating a high degree of radical stability as well as a robust resistance to the ionizing effects of gamma irradiation. The rationally designed melanins protected mammalian cells against ionizing radiation of different energies. We propose that due to melanin's numerous aromatic oligomers containing multiple pi-electron system, a generated Compton recoil electron gradually loses energy while passing through the pigment, until its energy is sufficiently low that it can be trapped by stable free radicals present in the pigment. Controlled dissipation of high-energy recoil electrons by melanin prevents secondary ionizations and the generation of damaging free radical species.

From Surface Chlorophyll a to Phytoplankton Community Composition in Oceanic Waters

NASA Technical Reports Server (NTRS)

Uitz, Julia; Claustre, Herve; Morel, Andre; Hooker, Stanford B.

2004-01-01

The objective of the present study is to examine the potential of using the near-surface total chlorophyll a concentration (C(sub surf)), as it can be derived from ocean color observation, to infer the column-integrated and the vertical distribution of the phytoplanktonic biomass, both in a quantitative way and in a qualitative way (z.e., in terms of community structure). Within this context, a large HPLC (High Performance Liquid Chromatography) pigment database has been analyzed. It includes 2419 vertical pigment profiles, all sampled in Case-1 waters with various trophic states. The relationshps between C(sub surf) and the total chlorophyll alpha vertical distribution, as previously derived by Morel and Berthon, are fully confirmed, as the present results coincide with the previous ones. This agreement allows to go further, namely to examine the possibility of extracting relationships between C(sub surf) and the vertical composition of the algal assemblages. Thanks to the detailed pigment composition available from HPLC measurements, the contribution of three size classes (micro-, nano-, and pico-phytoplankton) to the local total chlorophyll a concentration can be assessed. Corroborating previous findings (e.g., large species dominate in eutrophc environments, whereas tiny phytoplankton prevail in oligotrophic zones), the results lead to a statistically based parameterization. The predictive skill of this parameterization is successfully tested on a separate data set. With such a tool, the vertical total chlorophyll a profiles associated with each size class can be inferred from the sole knowledge of C(sub surf). By combining this tool with satellite ocean color data, it becomes conceivable to quantify on a global scale the phytoplankton biomass associated with each of the three size classes.

Rapid purification method for vitamin A-derived aging pigments A2E and iso-A2E using cation exchange resin.

PubMed

Jee, Eun Hye; Kim, So Ra; Jang, Young Pyo

2012-08-17

A2E, known to be involved in the pathogenesis of age-related macular degeneration (AMD), is one of the major compounds that accumulate as fluorescent pigments in retinal pigment epithelial (RPE) cells with age and in some retinal disorders. While the biomimetic synthesis of A2E and its cis-isomer, iso-A2E is as simple as 'one-pot' reaction, the purification of these amphiphillic compounds has been a bottleneck for the mass production of these pathophysiologically important eye pigments. In order to provide a new method of rapid purification of A2E and iso-A2E, we employed a cation exchange resin for the separation of these pigments from crude reaction mixture. The reaction mixture was loaded on a weak acid resin and was eluted with 80% methanol with sodium hydroxide (pH 12), 100% methanol, and 100% methanol with 0.1% trifluoroacetic acid (TFA) in sequence. A2E and isoA2E were eluted only with 100% methanol solution containing TFA. Most of unreacted starting materials and intermediates were removed with 80% methanol containing sodium hydroxide. The new method can be used as a relatively simple and economic way to purify A2E and iso-A2E compared to conventional HPLC technique. Copyright © 2012 Elsevier B.V. All rights reserved.

Enological Tannin Effect on Red Wine Color and Pigment Composition and Relevance of the Yeast Fermentation Products.

PubMed

García-Estévez, Ignacio; Alcalde-Eon, Cristina; Puente, Víctor; Escribano-Bailón, M Teresa

2017-11-23

Enological tannins are widely used in the winemaking process either to improve different wine characteristics (color stability, among others) or to compensate for low tannin levels. In this work, the influence of the addition of two different enological tannins, mainly composed of hydrolysable (ellagitannins) and condensed tannins, on the evolution of color and pigment composition of two different types of model systems containing the five main grape anthocyanins was studied. In addition, the effect of the addition of an enological tannin on the color and pigment composition of red wines made from Vitis vinifera L. cv Tempranillo grapes was also studied by high-performance liquid chromatography with diode array detection coupled to mass spectrometry (HPLC-DAD-MS). Results showed that, in model systems, the addition of the enological tannin favored the formation of anthocyanin-derived pigments, such as A-type and B-type vitisins and flavanol-anthocyanin condensation products, provided that the yeast precursors were previously supplied. Moreover, model systems containing the enological tannins were darker and showed higher values of chroma at the end of the study than control ones. The higher formation of these anthocyanin-derived pigments was also observed in the red wines containing the enological tannin. Moreover, these wine also showed lower lightness (L*) values and higher chroma (C* ab ) values than control wines, indicating a higher stabilization of color.

Variation of phytoplankton assemblages along the Mozambique coast as revealed by HPLC and microscopy

NASA Astrophysics Data System (ADS)

Sá, C.; Leal, M. C.; Silva, A.; Nordez, S.; André, E.; Paula, J.; Brotas, V.

2013-05-01

This study is an integrated overview of pigment and microscopic analysis of phytoplankton communities throughout the Mozambican coast. Collected samples revealed notable patterns of phytoplankton occurrence and distribution, with community structure changing between regions and sample depth. Pigment data showed Delagoa Bight, Sofala Bank and Angoche as the most productive regions throughout the sampled area. In general, micro-sized phytoplankton, particularly diatoms, were important contributors to biomass both at surface and sub-surface maximum (SSM) samples, although were almost absent in the northern stations. In contrast, nano- and pico-sized phytoplankton revealed opposing patterns. Picophytoplankton were most abundant at surface, as opposed to nanophytoplankton, which were more abundant at the SSM. Microphytoplankton were associated with cooler southern water masses, while picophytoplankton were related to warmer northern water masses. Nanophytoplankton were found to increase their contribution to biomass with increasing SSM. Microscopy information on the genera and species level revealed the diatoms Chaetoceros spp., Proboscia alata, Pseudo-nitzschia spp., Cylindrotheca closterium and Hemiaulus haukii as the most abundant taxa of the micro-sized phytoplankton. Discosphaera tubifera and Emiliania huxleyi were the most abundant coccolithophores, nano-sized phytoplankton.

[Content comparison of diterpenoid pigments in different processed products of Gardenia jasminoides].

PubMed

Zhang, Cun; Xiao, Yong-Qing; Li, Li; Li, Gui-Liu; Pang, Zhen

2008-11-01

To compare the contents of diterpenoid pigments in the different processed products of Gardenia jasminoides. The separation of Crocin 1, Crocin 2, Crocin 3, Crocetin were determined simultaneously by HPLC on a kromasil C18 column at 35 degrees C with the m methanol-acetonitrile-0.3% formic acid anhydrous in gradient elution as the mobile phrase. The detection wavelength was set at 440 nm and the flow rate was 1.0 mL x min(-1). The obtained linearity of the four components was better over 0.9995 and the average recoveries were 97.77%, 100.05% , 98.40%, 101.02%, respectively. The method is simple, accurate with good reproducibility. The results showed that the remarkable variation regulations appear among the different processed products.

Analysis and enhancement of astaxanthin accumulation in Haematococcus pluvialis.

PubMed

Orosa, M; Franqueira, D; Cid, A; Abalde, J

2005-02-01

The green microalga Haematococcus pluvialis was cultured with different concentrations of NaNO(3) to determine the effect on cell growth and astaxanthin accumulation. The optimum nitrate concentration to obtain astaxanthin and to avoid the cessation of cell division was 0.15 g/l NaNO(3). The ratio chlorophyll a/total carotenoids proved a good physiological indicator of nitrogen deficiency in the cell. The effect of different carbon sources, malonate and acetate, on astaxanthin accumulation was also studied; up to 13 times more carotenoids per cell were accumulated in cultures with malonate than in cultures without this compound. The pigment analysis was performed by a new low toxicity HPLC method capable of separating chlorophylls a and b, carotenes and xanthophylls in a short-period of time, using low volumes of solvents and with an economical price. With this method even echinenone was separated, which had been unsuccessful by any other method.

Spring temperature variability and eutrophication history inferred from sedimentary pigments in the varved sediments of Lake Żabińskie, north-eastern Poland, AD 1907-2008

NASA Astrophysics Data System (ADS)

Amann, Benjamin; Lobsiger, Simon; Fischer, Daniela; Tylmann, Wojciech; Bonk, Alicja; Filipiak, Janusz; Grosjean, Martin

2014-12-01

Varved lake sediments are excellent natural archives providing quantitative insights into climatic and environmental changes at very high resolution and chronological accuracy. However, due to the multitude of responses within lake ecosystems it is often difficult to understand how climate variability interacts with other environmental pressures such as eutrophication, and to attribute observed changes to specific causes. This is particularly challenging during the past 100 years when multiple strong trends are superposed. Here we present a high-resolution multi-proxy record of sedimentary pigments and other biogeochemical data from the varved sediments of Lake Żabińskie (Masurian Lake District, north-eastern Poland, 54°N-22°E, 120 m a.s.l.) spanning AD 1907 to 2008. Lake Żabińskie exhibits biogeochemical varves with highly organic late summer and winter layers separated by white layers of endogenous calcite precipitated in early summer. The aim of our study is to investigate whether climate-driven changes and anthropogenic changes can be separated in a multi-proxy sediment data set, and to explore which sediment proxies are potentially suitable for long quantitative climate reconstructions. We also test if convoluted analytical techniques (e.g. HPLC) can be substituted by rapid scanning techniques (visible reflectance spectroscopy VIS-RS; 380-730 nm). We used principal component analysis and cluster analysis to show that the recent eutrophication of Lake Żabińskie can be discriminated from climate-driven changes for the period AD 1907-2008. The eutrophication signal (PC1 = 46.4%; TOC, TN, TS, Phe-b, high TC/CD ratios total carotenoids/chlorophyll-a derivatives) is mainly expressed as increasing aquatic primary production, increasing hypolimnetic anoxia and a change in the algal community from green algae to blue-green algae. The proxies diagnostic for eutrophication show a smooth positive trend between 1907 and ca 1980 followed by a very rapid increase from ca. 1980 ± 2 onwards. We demonstrate that PC2 (24.4%, Chl-a-related pigments) is not affected by the eutrophication signal, but instead is sensitive to spring (MAM) temperature (r = 0.63, pcorr < 0.05, RMSEP = 0.56 °C; 5-yr filtered). Limnological monitoring data (2011-2013) support this finding. We also demonstrate that scanning visible reflectance spectroscopy (VIS-RS) data can be calibrated to HPLC-measured chloropigment data and be used to infer concentrations of sedimentary Chl-a derivatives {pheophytin a + pyropheophytin a}. This offers the possibility for very high-resolution (multi)millennial-long paleoenvironmental reconstructions.

Mycobacterium shottsii sp. nov., a slowly growing species isolated from Chesapeake Bay striped bass (Morone saxatilis)

USGS Publications Warehouse

Rhodes, M.W.; Kator, H.; Kotob, S.; van Berkum, P.; Kaattari, I.; Vogelbein, W.; Quinn, F.; Floyd, M.M.; Butler, W.R.; Ottinger, C.A.

2003-01-01

Slowly growing, non-pigmented mycobacteria were isolated from striped bass (Morone saxatilis) during an epizootic of mycobacteriosis in the Chesapeake Bay. Growth characteristics, acid-fastness and results of 16S rRNA gene sequencing were consistent with those of the genus Mycobacterium. A unique profile of biochemical reactions was observed among the 21 isolates. A single cluster of eight peaks identified by analysis of mycolic acids (HPLC) resembled those of reference patterns but differed in peak elution times from profiles of reference species of the Mycobacterium tuberculosis complex. One isolate (M175T) was placed within the slowly growing mycobacteria by analysis of aligned 16S rRNA gene sequences and was proximate in phylogeny to Mycobacterium ulcerans and Mycobacterium marinum. However, distinct nucleotide differences were detected in the 16S rRNA gene sequence among M175T, M. ulcerans and M. marinum (99.2% similarity). Isolate M175T could be differentiated from other slowly growing, non-pigmented mycobacteria by its inability to grow at 37??C, production of niacin and urease, absence of nitrate reductase and resistance to isoniazid (1 ??g ml-1), thiacetazone and thiophene-2-carboxylic hydrazide. Based upon these genetic and phenotypic differences, isolate M175T (= ATCC 700981T = NCTC 13215T) is proposed as the type strain of a novel species, Mycobacterium shottsii sp. nov.

Anthocyanins in the bracts of Curcuma species and relationship of the species based on anthocyanin composition.

PubMed

Koshioka, Masaji; Umegaki, Naoko; Boontiang, Kriangsuk; Pornchuti, Witayaporn; Thammasiri, Kanchit; Yamaguchi, Satoshi; Tatsuzawa, Fumi; Nakayama, Masayoshi; Tateishi, Akira; Kubota, Satoshi

2015-03-01

Five anthocyanins, delphinidin 3-O-rutinoside, cyanidin 3-O-rutinoside, petunidin 3-O-rutinoside, malvidin 3-O-glucoside and malvidin 3-O-rutinoside, were identified. Three anthocyanins, delphinidin 3-O-glucoside, cyanidin 3-O-glucoside and pelargonidin 3-O-rutinoside, were putatively identified based on C18 HPLC retention time, absorption spectrum, including λmax, and comparisons with those of corresponding standard anthocyanins, as the compounds responsible for the pink to purple-red pigmentation of the bracts of Curcuma alismatifolia and five related species. Cluster analysis based on four major anthocyanins formed two clusters. One consisted of only one species, C. alismatifolia, and the other consisted of five. Each cluster further formed sub-clusters depending on either species or habitats.

Quantification of active principles and pigments in leaf extracts of Isatis tinctoria by HPLC/UV/MS.

PubMed

Mohn, Tobias; Potterat, Olivier; Hamburger, Matthias

2007-02-01

An HPLC method has been developed and validated for the quantification of the pharmacologically active principles tryptanthrin (1), 1,3-dihydro-3-[(4-hydroxy-3,5-dimethoxyphenyl)methylene]-2 H-indol-2-one (indolinone) (3), indirubin (4), alpha-linolenic acid (2), and indigo (5), an isomer of indirubin, in extracts from the traditional anti-inflammatory plant Isatis tinctoria (woad). The chromatographic separation was performed on a C-18 column with a linear gradient of acetonitrile in water containing 0.1% formic acid. The method combines UV and electrospray MS detection in the positive ion mode for the detection of the alkaloids, with a switch to the negative mode for the analysis of alpha-linolenic acid. The method was applied to the analysis of woad extracts obtained by supercritical fluid (SFE) CO2 extraction, and by pressurized liquid extraction (PLE) with dichloromethane and methanol, respectively. While the highest concentration of alpha-linolenic acid was found in the SFE extract (7.43%), the concentrations of tryptanthrin , indolinone, indirubin and indigo were the highest in the dichloromethane extract (0.30, 0.035, 2.48 and 0.84%, respectively). Compound 3 was not detected in the methanolic extract and only traces of compounds 1, 4 and 5 and low amount of alpha-linolenic acid (0.39%) were present in this extract.

Tolerance of ARPE 19 cells to organophosphorus pesticide chlorpyrifos is limited to concentration and time of exposure.

PubMed

Gomathy, Narayanan; Sumantran, Venil N; Shabna, A; Sulochana, K N

2015-01-01

Age related macular degeneration is a blinding disease common in elder adults. The prevalence of age related macular degeneration has been found to be 1.8% in the Indian population. Organophosphates are widely used insecticides with well documented neurological effects, and the persistent nature of these compounds in the body results in long term health effects. Farmers exposed to organophosphorus pesticides in USA had an earlier onset of age related macular degeneration when compared to unexposed controls. A recent study found significant levels of an organophosphate, termed chlorpyrifos, in the blood samples of Indian farmers. Therefore, in understanding the link between age related macular degeneration and chlorpyrifos, the need for investigation is important. Our data show that ARPE-19 (retinal pigment epithelial cells) exhibit a cytoprotective response to chlorpyrifos as measured by viability, mitochondrial membrane potential, superoxide dismutase activity, and increased levels of glutathione peroxidase and reduced glutathione, after 24 h exposure to chlorpyrifos. However, this cytoprotective response was absent in ARPE-19 cells exposed to the same range of concentrations of chlorpyrifos for 48 h. These results have physiological significance, since HPLC analysis showed that effects of chlorpyrifos were mediated through its entry into ARPE-19 cells. HPLC analysis also showed that chlorpyrifos remained stable, as we recovered up to 80% of the chlorpyrifos added to 6 different ocular tissues. Copyright © 2014. Published by Elsevier Inc.

Rapid and simple immunochemical screening combined with hand-shaking extraction for thiamethoxam residue in agricultural products.

PubMed

Watanabe, Eiki; Kobara, Yuso; Miyake, Shiro

2013-06-01

With the aim of expanding the applicability of a kit-based enzyme-linked immunosorbent assay (ELISA) for the neonicotinoid insecticide thiamethoxam, the ELISA was newly applied to three kinds of agricultural samples (green pepper, eggplant and spinach). To offer the ELISA as a screening analysis for thiamethoxam residues, a rapid and simple method of extraction by hand-shaking was used, and speed-up and simplification of the sample treatment before the ELISA analysis were examined. Finally, the validity of the ELISA combined with the proposed extraction method was verified against a reference high-performance liquid chromatography (HPLC) method using real-world agricultural samples. There were no marked matrix effects derived from green pepper and eggplant extracts. On the other hand, although the effect due to a pigment in spinach extract on the assay performance was significant, it was effectively avoided by increasing the dilution level of the spinach extract. For thiamethoxam-spiked samples, acceptable recoveries of 97.9-109.1% and coefficients of variation of 0.3-11.5% were obtained. Inspection of the validity of the ELISA by comparison with the reference HPLC method showed that the two analytical results were very similar, and a high correlation was found between them (r>0.997). The evaluated ELISA combined with hand-shaking extraction provided a rapid and simple screening analysis that was quantitative and reliable for the detection of thiamethoxam in complex agricultural products. © 2012 Society of Chemical Industry.

Anthocyanins from purple sweet potato (Ipomoea batatas (L.) Lam.) and their color modulation by the addition of phenolic acids and food-grade phenolic plant extracts.

PubMed

Gras, Claudia C; Nemetz, Nicole; Carle, Reinhold; Schweiggert, Ralf M

2017-11-15

Anthocyanin profiles and contents of three purple sweet potato provenances were investigated by HPLC-DAD-MS n . In contrast to widely uniform profiles, the contents of total (558-2477mg/100gDM) and individual anthocyanins varied widely. Furthermore, quantitative and qualitative effects of intermolecular co-pigmentation were studied by adding chlorogenic and rosmarinic acids, and food-grade phenolic apple and rosemary extracts at various dosages to a diluted purple sweet potato concentrate at pH 0.9, 2.6, 3.6, and 4.6. Addition of co-pigments generally increased pK H estimate -values of anthocyanins from 3.28 (without co-pigments) to up to 4.71, thus substantially broadening the pH range wherein colored forms prevail. The most pronounced hyperchromic shift by up to +50.5% at the absorption maximum was observed at pH 4.6. Simply by blending the co-pigments with purple sweet potato anthocyanins at pH-values ranging from 2.6 to 4.6, purplish-blue, light pink, magenta, brick-red, and intense red hues were accessible as expressed by CIE-L ∗ a ∗ b ∗ color values. Copyright © 2017 Elsevier Ltd. All rights reserved.

Isolation of 4,5-O-Dicaffeoylquinic Acid as a Pigmentation Inhibitor Occurring in Artemisia capillaris Thunberg and Its Validation In Vivo.

PubMed

Tabassum, Nadia; Lee, Ji-Hyung; Yim, Soon-Ho; Batkhuu, Galzad Javzan; Jung, Da-Woon; Williams, Darren R

2016-01-01

There is a continual need to develop novel and effective melanogenesis inhibitors for the prevention of hyperpigmentation disorders. The plant Artemisia capillaris Thunberg (Oriental Wormwood) was screened for antipigmentation activity using murine cultured cells (B16-F10 malignant melanocytes). Activity-based fractionation using HPLC and NMR analyses identified the compound 4,5-O-dicaffeoylquinic acid as an active component in this plant. 4,5-O-Dicaffeoylquinic acid significantly reduced melanin synthesis and tyrosinase activity in a dose-dependent manner in the melanocytes. In addition, 4,5-O-dicaffeoylquinic acid treatment reduced the expression of tyrosinase-related protein-1. Significantly, we could validate the antipigmentation activity of this compound in vivo, using a zebrafish model. Moreover, 4,5-O-dicaffeoylquinic acid did not show toxicity in this animal model. Our discovery of 4,5-O-dicaffeoylquinic acid as an inhibitor of pigmentation that is active in vivo shows that this compound can be developed as an active component for formulations to treat pigmentation disorders.

Ultrasensitive and high-throughput analysis of chlorophyll a in marine phytoplankton extracts using a fluorescence microplate reader.

PubMed

Mandalakis, Manolis; Stravinskaitė, Austėja; Lagaria, Anna; Psarra, Stella; Polymenakou, Paraskevi

2017-07-01

Chlorophyll a (Chl a) is the predominant pigment in every single photosynthesizing organism including phytoplankton and one of the most commonly measured water quality parameters. Various methods are available for Chl a analysis, but the majority of them are of limited throughput and require considerable effort and time from the operator. The present study describes a high-throughput, microplate-based fluorometric assay for rapid quantification of Chl a in phytoplankton extracts. Microplate sealing combined with ice cooling was proved an effective means for diminishing solvent evaporation during sample loading and minimized the analytical errors involved in Chl a measurements with a fluorescence microplate reader. A set of operating parameters (settling time, detector gain, sample volume) were also optimized to further improve the intensity and reproducibility of Chl a fluorescence signal. A quadratic regression model provided the best fit (r 2  = 0.9998) across the entire calibration range (0.05-240 pg μL -1 ). The method offered excellent intra- and interday precision (% RSD 2.2 to 11.2%) and accuracy (% relative error -3.8 to 13.8%), while it presented particularly low limits of detection (0.044 pg μL -1 ) and quantification (0.132 pg μL -1 ). The present assay was successfully applied on marine phytoplankton extracts, and the overall results were consistent (average % relative error -14.8%) with Chl a concentrations (including divinyl Chl a) measured by high-performance liquid chromatography (HPLC). More importantly, the microplate-based method allowed the analysis of 96 samples/standards within a few minutes, instead of hours or days, when using a traditional cuvette-based fluorometer or an HPLC system. Graphical abstract TChl a concentrations (i.e. sum of Chl a and divinyl Chl a in ng L -1 ) measured in seawater samples by HPLC and fluorescence microplate reader.

Tropical bat as mammalian model for skin carotenoid metabolism

PubMed Central

Galván, Ismael; Garrido-Fernández, Juan; Ríos, José; Pérez-Gálvez, Antonio; Rodríguez-Herrera, Bernal

2016-01-01

Animals cannot synthesize carotenoid pigments de novo, and must consume them in their diet. Most mammals, including humans, are indiscriminate accumulators of carotenoids but inefficiently distribute them to some tissues and organs, such as skin. This limits the potential capacity of these organisms to benefit from the antioxidant and immunostimulatory functions that carotenoids fulfill. Indeed, to date, no mammal has been known to have evolved physiological mechanisms to incorporate and deposit carotenoids in the skin or hair, and mammals have therefore been assumed to rely entirely on other pigments such as melanins to color their integument. Here we use high-performance liquid chromatography (HPLC) in combination with time-of-flight mass spectrometry (HPLC-TOF/MS) to show that the frugivorous Honduran white bat Ectophylla alba colors its skin bright yellow with the deposition of the xanthophyll lutein. The Honduran white bat is thus a mammalian model that may help developing strategies to improve the assimilation of lutein in humans to avoid macular degeneration. This represents a change of paradigm in animal physiology showing that some mammals actually have the capacity to accumulate dietary carotenoids in the integument. In addition, we have also discovered that the majority of the lutein in the skin of Honduran white bats is present in esterified form with fatty acids, thereby permitting longer-lasting coloration and suggesting bright color traits may have an overlooked role in the visual communication of bats. PMID:27621447

The effect of blue light on periodontal biofilm growth in vitro.

PubMed

Fontana, Carla R; Song, Xiaoqing; Polymeri, Angeliki; Goodson, J Max; Wang, Xiaoshan; Soukos, Nikolaos S

2015-11-01

We have previously shown that blue light eliminates the black-pigmented oral bacteria Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, and Prevotella melaninogenica. In the present study, the in vitro photosensitivity of the above black-pigmented microorganisms and four Fusobacteria species (Fusobacterium nucleatum ss. nucleatum, F. nucleatum ss. vincentii, F. nucleatum ss. polymorphum, Fusobacterium periodonticum) was investigated in pure cultures and human dental plaque suspensions. We also tested the hypothesis that phototargeting the above eight key periodontopathogens in plaque-derived biofilms in vitro would control growth within the dental biofilm environment. Cultures of the eight bacteria were exposed to blue light at 455 nm with power density of 80 mW/cm2 and energy fluence of 4.8 J/cm2. High-performance liquid chromatography (HPLC) analysis of bacteria was performed to demonstrate the presence and amounts of porphyrin molecules within microorganisms. Suspensions of human dental plaque bacteria were also exposed once to blue light at 455 nm with power density of 50 mW/cm2 and energy fluence of 12 J/cm2. Microbial biofilms developed from the same plaque were exposed to 455 nm blue light at 50 mW/cm2 once daily for 4 min (12 J/cm2) over a period of 3 days (4 exposures) in order to investigate the cumulative action of phototherapy on the eight photosensitive pathogens as well as on biofilm growth. Bacterial growth was evaluated using the colony-forming unit (CFU) assay. The selective phototargeting of pathogens was studied using whole genomic probes in the checkerboard DNA-DNA format. In cultures, all eight species showed significant growth reduction (p < 0.05). HPLC demonstrated various porphyrin patterns and amounts of porphyrins in bacteria. Following phototherapy, the mean survival fractions were reduced by 28.5 and 48.2% in plaque suspensions and biofilms, respectively, (p < 0.05). DNA probe analysis showed significant reduction in relative abundances of the eight bacteria as a group in plaque suspensions and biofilms. The cumulative blue light treatment suppressed biofilm growth in vitro. This may introduce a new avenue of prophylactic treatment for periodontal diseases.

Isolation and antisense suppression of flavonoid 3', 5'-hydroxylase modifies flower pigments and colour in cyclamen.

PubMed

Boase, Murray R; Lewis, David H; Davies, Kevin M; Marshall, Gayle B; Patel, Deepa; Schwinn, Kathy E; Deroles, Simon C

2010-06-13

Cyclamen is a popular and economically significant pot plant crop in several countries. Molecular breeding technologies provide opportunities to metabolically engineer the well-characterized flavonoid biosynthetic pathway for altered anthocyanin profile and hence the colour of the flower. Previously we reported on a genetic transformation system for cyclamen. Our aim in this study was to change pigment profiles and flower colours in cyclamen through the suppression of flavonoid 3', 5'-hydroxylase, an enzyme in the flavonoid pathway that plays a determining role in the colour of anthocyanin pigments. A full-length cDNA putatively identified as a F3'5'H (CpF3'5'H) was isolated from cyclamen flower tissue. Amino acid and phylogeny analyses indicated the CpF3'5'H encodes a F3'5'H enzyme. Two cultivars of minicyclamen were transformed via Agrobacterium tumefaciens with an antisense CpF3'5'H construct. Flowers of the transgenic lines showed modified colour and this correlated positively with the loss of endogenous F3'5'H transcript. Changes in observed colour were confirmed by colorimeter measurements, with an overall loss in intensity of colour (C) in the transgenic lines and a shift in hue from purple to red/pink in one cultivar. HPLC analysis showed that delphinidin-derived pigment levels were reduced in transgenic lines relative to control lines while the percentage of cyanidin-derived pigments increased. Total anthocyanin concentration was reduced up to 80% in some transgenic lines and a smaller increase in flavonol concentration was recorded. Differences were also seen in the ratio of flavonol types that accumulated. To our knowledge this is the first report of genetic modification of the anthocyanin pathway in the commercially important species cyclamen. The effects of suppressing a key enzyme, F3'5'H, were wide ranging, extending from anthocyanins to other branches of the flavonoid pathway. The results illustrate the complexity involved in modifying a biosynthetic pathway with multiple branch points to different end products and provides important information for future flower colour modification experiments in cyclamen.

Isolation and antisense suppression of flavonoid 3', 5'-hydroxylase modifies flower pigments and colour in cyclamen

PubMed Central

2010-01-01

Background Cyclamen is a popular and economically significant pot plant crop in several countries. Molecular breeding technologies provide opportunities to metabolically engineer the well-characterized flavonoid biosynthetic pathway for altered anthocyanin profile and hence the colour of the flower. Previously we reported on a genetic transformation system for cyclamen. Our aim in this study was to change pigment profiles and flower colours in cyclamen through the suppression of flavonoid 3', 5'-hydroxylase, an enzyme in the flavonoid pathway that plays a determining role in the colour of anthocyanin pigments. Results A full-length cDNA putatively identified as a F3'5'H (CpF3'5'H) was isolated from cyclamen flower tissue. Amino acid and phylogeny analyses indicated the CpF3'5'H encodes a F3'5'H enzyme. Two cultivars of minicyclamen were transformed via Agrobacterium tumefaciens with an antisense CpF3'5'H construct. Flowers of the transgenic lines showed modified colour and this correlated positively with the loss of endogenous F3'5'H transcript. Changes in observed colour were confirmed by colorimeter measurements, with an overall loss in intensity of colour (C) in the transgenic lines and a shift in hue from purple to red/pink in one cultivar. HPLC analysis showed that delphinidin-derived pigment levels were reduced in transgenic lines relative to control lines while the percentage of cyanidin-derived pigments increased. Total anthocyanin concentration was reduced up to 80% in some transgenic lines and a smaller increase in flavonol concentration was recorded. Differences were also seen in the ratio of flavonol types that accumulated. Conclusion To our knowledge this is the first report of genetic modification of the anthocyanin pathway in the commercially important species cyclamen. The effects of suppressing a key enzyme, F3'5'H, were wide ranging, extending from anthocyanins to other branches of the flavonoid pathway. The results illustrate the complexity involved in modifying a biosynthetic pathway with multiple branch points to different end products and provides important information for future flower colour modification experiments in cyclamen. PMID:20540805

High biological variability of plastids, photosynthetic pigments and pigment forms of leaf primordia in buds.

PubMed

Solymosi, Katalin; Morandi, Dominique; Bóka, Károly; Böddi, Béla; Schoefs, Benoît

2012-05-01

To study the formation of the photosynthetic apparatus in nature, the carotenoid and chlorophyllous pigment compositions of differently developed leaf primordia in closed and opening buds of common ash (Fraxinus excelsior L.) and horse chestnut (Aesculus hippocastanum L.) as well as in closed buds of tree of heaven (Ailanthus altissima P. Mill.) were analyzed with HPLC. The native organization of the chlorophyllous pigments was studied using 77 K fluorescence spectroscopy, and plastid ultrastructure was investigated with electron microscopy. Complete etiolation, i.e., accumulation of protochlorophyllide, and absence of chlorophylls occurred in the innermost leaf primordia of common ash buds. The other leaf primordia were partially etiolated in the buds and contained protochlorophyllide (0.5-1 μg g(-1) fresh mass), chlorophyllides (0.2-27 μg g(-1) fresh mass) and chlorophylls (0.9-643 μg g(-1) fresh mass). Etio-chloroplasts with prolamellar bodies and either regular or only low grana were found in leaves having high or low amounts of chlorophyll a and b, respectively. After bud break, etioplast-chloroplast conversion proceeded and the pigment contents increased in the leaves, similarly to the greening processes observed in illuminated etiolated seedlings under laboratory conditions. The pigment contents and the ratio of the different spectral forms had a high biological variability that could be attributed to (i) various light conditions due to light filtering in the buds resulting in differently etiolated leaf primordia, (ii) to differences in the light-exposed and inner regions of the same primordia in opening buds due to various leaf folding, and (iii) to tissue-specific slight variations of plastid ultrastructure.

Hydrolysis kinetics of astaxanthin esters and stability of astaxanthin of Haematococcus pluvialis during saponification.

PubMed

Yuan, J P; Chen, F

1999-01-01

The reaction kinetics for the hydrolysis of astaxanthin esters and the degradation of astaxanthin during saponification of the pigment extract from the microalga Haematococcus pluvialis were investigated. Different concentrations of sodium hydroxide in methanol were used for the saponification under nitrogen in darkness at ambient temperature (22 degrees C) followed by the analysis of astaxanthins and other carotenoids using an HPLC method. The concentration of methanolic NaOH solution was important for promoting the hydrolysis of astaxanthin esters and minimizing the degradation of astaxanthin during saponification. With a higher concentration of methanolic NaOH solution, the reaction rate of hydrolysis was high, but the degradation of astaxanthin occurred significantly. The rate constants of the hydrolysis reaction (first order) of astaxanthin esters and the degradation reaction (zero-order) of astaxanthin were directly proportional to the concentration of sodium hydroxide in the saponified solution. Although the concentration of sodium hydroxide in the saponified solution was 0.018 M, complete hydrolysis of astaxanthin esters was achieved in 6 h for different concentrations (10-100 mg/L) of pigment extracts. Results also indicated that a higher temperature should be avoided to minimize the degradation of astaxanthin. In addition, during saponification, no loss of lutein, beta-carotene, and canthaxanthin was found.

Transformation of algal turf by echinoids and scarid fishes on French Polynesian coral reefs

NASA Astrophysics Data System (ADS)

Harmelin-Vivien, Mireille L.; Peyrot-Clausade, Mireille; Romano, Jean-Claude

1992-04-01

The respective roles of regular echinoids and scarid fishes in the transformation of turf algae, the main food resource for reef herbivores, were investigated on French Polynesian coral reefs. The role of one species of parrotfish ( Scarus sordidus) was compared with that of four species of echinoids. The degree and ways of degradation of the algal matter were determined by the organic matter percentage, the composition of the sugar fraction, and the concentration and composition of chlorophylltype pigments as assayed by HPLC analysis. Chemical analyses were performed on anterior and posterior intestines for scarids, intestinal contents and faeces for echinoids, and on fresh algal turf as a control of initial food quality. A decrease in mean percentage of organic matter in gut content was observed from intestine (9.7%) to faeces (7%) in sea urchins, but not in parrotfishes. The total sugar fraction decreased from fresh algal turf (32% of total organic matter) to echinoid (28%) to scarid (18%) gut contents. The ratio of insoluble to soluble sugars (I/S ratios) was higher in echinoids (2.6) than in scarid gut contents (1.0). A decrease in the total pigment concentration was measured from fresh algal turf to echinoid and it was found to be even lower in scarid gut contents. Chromatograms showed that the composition of chlorophyll-type pigments in scarid intestines was very similar to fresh algal turf, with a dominance of native forms, mainly chlorophyll a and b. On the contrary, degraded pigment forms dominated in echinoids. The main degraded products were pheophorbides in sea urchins, and chlorophyllides in parrotfishes. These results provided evidence for differentiation in digestive processes occurring in the two types of grazers. Echinoids released higher degraded algal material than did scarids. Thus, these two types of grazers play different roles in the recycling of organic matter on coral reefs.

Comparative Study of Antioxidant Power, Polyphenols, Flavonoids and Betacyanins of the Peel and Pulp of Three Tunisian Opuntia Forms

PubMed Central

Yeddes, Nizar; Chérif, Jamila K.; Guyot, Sylvain; Sotin, Hélène; Ayadi, Malika T.

2013-01-01

The antioxidant activity and the chemical composition of methanol extracts from peel and pulp belonging to two species of Tunisian prickly pears Opuntia ficus indica (spiny and thornless forms) and Opuntia stricta have been studied. The antioxidant capacity was measured by DPPH radical scavenging activity. The total phenolic compound (TPC) and the total flavonoid content were determined by the Folin–Ciocalteu method and colorimetric method, respectively. The phenolic compounds were identified and quantified by high-performance liquid chromatography (HPLC) coupled with an electrospray ionization mass spectrometry (ESI-MS). The results showed that O. stricta fruits present the best antioxidant activities than the two forms of O. ficus indica, while the TPC was more important in O. ficus indica than in the O. stricta fruits. The peels have higher flavonoids than pulp, and the thornless variety has more flavonoid than the spiny. The RP-HPLC and ESI-MS analysis detected two classes of phenolic compounds and betalain pigments. Isorhamnetin derivatives are the dominant flavonol glycoside identified in O. ficus indica (spiny: 65.25 μg·g−1; thornless: 77.03 μg·g−1) and O. stricta peels (19.22 μg·g−1). PMID:26787622

Investigation of genetic variation in scavenger receptor class B, member 1 (SCARB1) and association with serum carotenoids

PubMed Central

McKay, Gareth J; Loane, Edward; Nolan, John M; Patterson, Christopher C; Meyers, Kristin J; Mares, Julie A; Yonova-Doing, Ekaterina; Hammond, Christopher J; Beatty, Stephen; Silvestri, Giuliana

2013-01-01

Objective To investigate association of scavenger receptor class B, member 1 (SCARB1) genetic variants with serum carotenoid levels of lutein (L) and zeaxanthin (Z) and macular pigment optical density (MPOD). Design A cross-sectional study of healthy adults aged 20-70. Participants 302 participants recruited following local advertisement. Methods MPOD was measured by customized heterochromatic flicker photometry. Fasting blood samples were taken for serum L and Z measurement by HPLC and lipoprotein analysis by spectrophotometric assay. Forty-seven single nucleotide polymorphisms (SNPs) across SCARB1 were genotyped using Sequenom technology. Association analyses were performed using PLINK to compare allele and haplotype means, with adjustment for potential confounding and correction for multiple comparisons by permutation testing. Replication analysis was performed in the TwinsUK and CAREDS cohorts. Main outcome measures Odds ratios (ORs) for macular pigment optical density area, serum lutein and zeaxanthin concentrations associated with genetic variations in SCARB1 and interactions between SCARB1 and sex. Results Following multiple regression analysis with adjustment for age, body mass index, sex, high-density lipoprotein cholesterol (HDLc), low-density lipoprotein cholesterol (LDLc), triglycerides, smoking, dietary L and Z levels, 5 SNPs were significantly associated with serum L concentration and 1 SNP with MPOD (P<0.01). Only the association between rs11057841 and serum L withstood correction for multiple comparisons by permutation testing (P<0.01) and replicated in the TwinsUK cohort (P=0.014). Independent replication was also observed in the CAREDS cohort with rs10846744 (P=2×10−4), a SNP in high linkage disequilibrium with rs11057841 (r2=0.93). No significant interactions by sex were found. Haplotype analysis revealed no stronger association than obtained with single SNP analyses. Conclusions Our study has identified association between rs11057841 and serum L concentration (24% increase per T allele) in healthy subjects, independent of potential confounding factors. Our data supports further evaluation of the role for SCARB1 in the transport of macular pigment and the possible modulation of AMD risk through combating the effects of oxidative stress within the retina. PMID:23562302

Spiroketalcarminic Acid, a Novel Minor Anthraquinone Pigment in Cochineal Extract Used in Food Additives.

PubMed

Ito, Yusai; Harikai, Naoki; Ishizuki, Kyoko; Shinomiya, Kazufusa; Sugimoto, Naoki; Akiyama, Hiroshi

2017-09-01

Cochineal extract prepared from the scale insect Dactylopus coccus (American cochineal) has been used as a natural red dye for food, cosmetics, and pharmaceuticals. The major pigment in cochineal extract is carminic acid (CA), an anthraquinone glucoside, and several minor pigments have been previously reported. Our investigation aimed at establishing the safety of cochineal dye products using ultra performance liquid chromatography-photo diode array-electrospray ionization-time of flight (UPLC-PDA-ESI-TOF)/MS found an unknown minor pigment, spiroketalcarminic acid (1), in three commercial cochineal extract samples; cochineal extract used in food additives, carmine that is an aluminum salt of cochineal extract used as natural dye, and a research reagent of CA. The purification of 1 from cochineal extract involved sequential chromatographic techniques, including preparative reversed-phase HPLC. Two dimensional (2D)-NMR and mass analyses established the structure of 1 to be a novel anthraquinone with an unusual 6,5-spiroketal system instead of the C-glucosyl moiety of CA. The absolute stereochemistry of the spiroketal moiety in 1 was determined by nuclear Overhauser effect spectroscopy (NOESY) correlations and optical rotation. No data corresponding to 1 had previously been reported for extracts of dried cochineal insects and traditional art products dyed with cochineal extract, indicating that 1 is likely produced during the preparation of commercial cochineal extract.

Comparison of Aquitaine and Rioja Red Wines: Characterization of Their Phenolic Composition and Evolution from 2000 to 2013.

PubMed

Quaglieri, Cindy; Prieto-Perea, Noelia; Berrueta, Luis Angel; Gallo, Blanca; Rasines-Perea, Zurine; Jourdes, Michael; Teissedre, Pierre-Louis

2017-01-24

Wine chemical analysis was carried out on 194 commercial blended red wines produced by two major wine-growing areas-the Aquitaine (France) and Rioja (Spain) regions-in order to compare the wines of both regions. Anthocyanins and derived pigments, tannins and derivatives were identified and quantified by HPLC-DAD-ESI-MS/MS (high pressure liquid chromatography coupled to diode array detector and mass spectrometry using the electrospray ionization interface). Mean degree of polymerization (mDP) was determined. The influence of the wine-growing region and the predominance of the properties of some grape varieties used are confirmed by the significant differences observed between both regions. Rioja and Bordeaux "generic" (Bordeaux and Bordeaux-Supérieur appellations) red wines showed the highest anthocyanic content and the highest mDP, as these wines are in a majority made from Merlot (Bordeaux "generic") and Tempranillo (Rioja). On the contrary, Bordeaux "specific" regions (Blayais, Médoc, Graves, and Libournais) showed the red wines with the highest total phenolic content and tannin concentration, as the predominant grape variety used is Cabernet Sauvignon. A principal component analysis (PCA) and a hierarchical ascendant classification (HAC) suggesting patterns between the chemical parameters and the distribution of the red wines in three groups were proposed. The comparison of the two wine-growing areas also reveals some similarities between the various grape varieties used. A general effect of a progressive decrease in anthocyanins, anthocyanin-derived pigment and tannins is observed for older wines.

Production of red-flowered plants by genetic engineering of multiple flavonoid biosynthetic genes.

PubMed

Nakatsuka, Takashi; Abe, Yoshiko; Kakizaki, Yuko; Yamamura, Saburo; Nishihara, Masahiro

2007-11-01

Orange- to red-colored flowers are difficult to produce by conventional breeding techniques in some floricultural plants. This is due to the deficiency in the formation of pelargonidin, which confers orange to red colors, in their flowers. Previous researchers have reported that brick-red colored flowers can be produced by introducing a foreign dihydroflavonol 4-reductase (DFR) with different substrate specificity in Petunia hybrida, which does not accumulate pelargonidin pigments naturally. However, because these experiments used dihydrokaempferol (DHK)-accumulated mutants as transformation hosts, this strategy cannot be applied directly to other floricultural plants. Thus in this study, we attempted to produce red-flowered plants by suppressing two endogenous genes and expressing one foreign gene using tobacco as a model plant. We used a chimeric RNAi construct for suppression of two genes (flavonol synthase [FLS] and flavonoid 3'-hydroxylase [F3'H]) and expression of the gerbera DFR gene in order to accumulate pelargonidin pigments in tobacco flowers. We successfully produced red-flowered tobacco plants containing high amounts of additional pelargonidin as confirmed by HPLC analysis. The flavonol content was reduced in the transgenic plants as expected, although complete inhibition was not achieved. Expression analysis also showed that reduction of the two-targeted genes and expression of the foreign gene occurred simultaneously. These results demonstrate that flower color modification can be achieved by multiple gene regulation without use of mutants if the vector constructs are designed resourcefully.

Effect of fining on the colour and pigment composition of young red wines.

PubMed

González-Neves, Gustavo; Favre, Guzmán; Gil, Graciela

2014-08-15

This work aimed to evaluate the effect of four fining agents on the colour and pigment composition of red wines of Tannat. The wines were analysed 15 days after fining and immediately after separation of sediments and bottling. Colour was evaluated by spectrophotometry and polyphenols were analysed by spectrophotometry and HPLC-DAD. The colour intensity of wine was significantly decreased by bentonite and egg albumin. The most remarkable effects on wine phenolic composition were produced by bentonite and gelatin, which significantly decreased anthocyanin and tannin concentrations, respectively. Results show that each fining agent has very different impact on the wine attributes, and their effects depended as well on the composition of the clarified wine. The use of non-traditional agents of fining, as vegetable proteins, may have less impact on the colour and anthocyanin content of red wines. Copyright © 2014 Elsevier Ltd. All rights reserved.

Measurement of food flavonoids by high-performance liquid chromatography: A review.

PubMed

Merken, H M; Beecher, G R

2000-03-01

The flavonoids are plant polyphenols found frequently in fruits, vegetables, and grains. Divided into several subclasses, they include the anthocyanidins, pigments chiefly responsible for the red and blue colors in fruits, fruit juices, wines, and flowers; the catechins, concentrated in tea; the flavanones and flavanone glycosides, found in citrus and honey; and the flavones, flavonols, and flavonol glycosides, found in tea, fruits, vegetables, and honey. Known for their hydrogen-donating antioxidant activity as well as their ability to complex divalent transition metal cations, flavonoids are propitious to human health. Computer-controlled high-performance liquid chromatography (HPLC) has become the analytical method of choice. Many systems have been developed for the detection and quantification of flavonoids across one, two, or three subclasses. A summary of the various HPLC and sample preparation methods that have been employed to quantify individual flavonoids within a subclass or across several subclasses are tabulated in this review.

Cytokinin delays dark-induced senescence in rice by maintaining the chlorophyll cycle and photosynthetic complexes.

PubMed

Talla, Sai Krishna; Panigrahy, Madhusmita; Kappara, Saivishnupriya; Nirosha, P; Neelamraju, Sarla; Ramanan, Rajeshwari

2016-03-01

The phytohormone cytokinin (CK) is known to delay senescence in plants. We studied the effect of a CK analog, 6-benzyl adenine (BA), on rice leaves to understand the possible mechanism by which CK delays senescence in a drought- and heat-tolerant rice cultivar Nagina22 (N22) using dark-induced senescence (DIS) as a surrogate for natural senescence of leaves. Leaves of N22-H-dgl162, a stay-green mutant of N22, and BA-treated N22 showed retention of chlorophyll (Chl) pigments, maintenance of the Chl a/b ratio, and delay in reduction of both photochemical efficiency and rate of oxygen evolution during DIS. HPLC analysis showed accumulation of 7-hydroxymethyl chlorophyll (HmChl) during DIS, and the kinetics of its accumulation correlated with progression of senescence. Transcriptome analysis revealed that several plastid-localized genes, specifically those associated with photosystem II (PSII), showed higher transcript levels in BA-treated N22 and the stay-green mutant leaves compared with naturally senescing N22 leaves. Real-time PCR analyses showed that genes coding for enzymes associated with Chl a/b interconversion and proteins associated with light-harvesting complexes maintained higher transcript levels up to 72h of DIS following BA treatment. The pigment-protein complexes analyzed by green gel remained intact in both N22-H-dgl162 and BA-treated N22 leaves even after 96h of DIS. Thus, CK delays senescence by accumulation of HmChl and up-regulating genes in the Chl cycle, thereby maintaining the Chl a/b ratio. Also, CK treatment retains higher transcript levels of PSII-related genes, resulting in the stability of photosynthetic pigment complexes and functional stay-greenness in rice. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Finding of 13(2) , 17(3) -cyclopheophorbide a enol as a degradation product of chlorophyll in shrunk zooxanthellae of the coral Montipora digitata.

PubMed

Suzuki, Toshiyuki; Casareto, Beatriz Estela; Shioi, Yuzo; Ishikawa, Yoshio; Suzuki, Yoshimi

2015-02-01

We examined the morphology and pigment composition of zooxanthellae in corals subjected to normal temperature (27°C) and thermal stress (32°C). We observed several normal and abnormal morphological types of zooxanthellar cells. Normal cells were intact and their chloroplasts were unbroken (healthy); abnormal cells were shrunken and had partially degraded or broken chloroplasts, or they were bleached and without chloroplasts. At 27°C, most healthy zooxanthellar cells were retained in the coral tissue, whereas shrunken zooxanthellae were expelled. Under thermal stress, the abundance of healthy zooxanthellae declined and the proportion of shrunken/abnormal cells increased in coral tissues. The rate of algal cell expulsion was reduced under thermal stress. Within the shrunken cells, we detected the presence of a chl-like pigment that is not ordinarily found in healthy zooxanthellae. Analysis of the absorption spectrum, absorption maxima, and retention time (by HPLC) indicated that this pigment was 13(2) , 17(3) -cyclopheophorbide a enol (cPPB-aE), which is frequently found in marine and lacustrine sediments, and in protozoans that graze on phytoplankton. The production of cPPB-aE in shrunken zooxanthellae suggests that the chls have been degraded to cPPB-aE, a compound that is not fluorescent. The lack of a fluorescence function precludes the formation of reactive oxygen species. We therefore consider the formation of cPPB-aE in shrunken zooxanthellae to be a mechanism for avoiding oxidative stress. © 2014 The Authors. Journal of Phycology published by Wiley Periodicals, Inc. on behalf of Phycological Society of America.

The Sea Urchin Arbacia lixula: A Novel Natural Source of Astaxanthin

PubMed Central

Cirino, Paola; Brunet, Christophe; Ciaravolo, Martina; Galasso, Christian; Musco, Luigi; Vega Fernández, Tomás; Sansone, Clementina; Toscano, Alfonso

2017-01-01

Several echinoderms, including sea urchins, are valuable sources of bioactive compounds but their nutraceutical potential is largely unexplored. In fact, the gonads of some sea urchin species contain antioxidants including carotenoids and polyhydroxylated naphthoquinones (PHNQ’s), such as echinochrome A. Astaxanthin is known to have particular bioactivity for the prevention of neurodegenerative diseases. This carotenoid is produced by microalgae, while several marine invertebrates can bioaccumulate or synthetize it from metabolic precursors. We determined the carotenoid content and analyzed the bioactivity potential of non-harvested Atlantic-Mediterranean sea urchin Arbacia lixula. The comparison of methanol crude extracts obtained from eggs of farmed and wild specimens revealed a higher bioactivity in farmed individuals fed with a customized fodder. HPLC-analysis revealed a high concentration of astaxanthin (27.0 μg/mg), which was the only pigment observed. This study highlights the potential of farmed A. lixula as a new source of the active stereoisomer of astaxanthin. PMID:28635649

Carotenoid Profile, Antioxidant Capacity, and Chromoplasts of Pink Guava (Psidium guajava L. Cv. 'Criolla') during Fruit Ripening.

PubMed

Rojas-Garbanzo, Carolina; Gleichenhagen, Maike; Heller, Annerose; Esquivel, Patricia; Schulze-Kaysers, Nadine; Schieber, Andreas

2017-05-10

Pigments of pericarp and pulp of pink guava (Psidium guajava L. cv. 'Criolla') were investigated to elucidate the profile and the accumulation of main carotenoids during four stages of fruit ripening by using HPLC-DAD and APCI-MS/MS analysis. Seventeen carotenoids were identified, and changes in their profile during fruit ripening were observed. The carotenoids all-trans-β-carotene, 15-cis-lycopene, and all-trans-lycopene were present in all ripening stages, but all-trans-lycopene was found to be predominant (from 63% to 92% of total carotenoids) and responsible for the high lipophilic antioxidant capacity determined by spectrophotometric assays. By using light and transmission electron microscopy, the development of chromoplasts in pericarp and pulp was demonstrated. The accumulation of all-trans-lycopene and all-trans-β-carotene coincided with the development of large crystals; the chromoplasts of pink guava belong, therefore, to the crystalline type.

UV-visible scanning spectrophotometry and chemometric analysis as tools for carotenoids analysis in cassava genotypes (Manihot esculenta Crantz).

PubMed

Moresco, Rodolfo; Uarrota, Virgílio Gavicho; Pereira, Aline; Tomazzoli, Maíra Maciel; Nunes, Eduardo da C; Peruch, Luiz Augusto Martins; Gazzola, Jussara; Costa, Christopher; Rocha, Miguel; Maraschin, Marcelo

2015-10-21

In this study, the metabolomics characterization focusing on the carotenoid composition of ten cassava (Manihot esculenta) genotypes cultivated in southern Brazil by UV-visible scanning spectrophotometry and reverse phase-high performance liquid chromatography was performed. Cassava roots rich in β-carotene are an important staple food for populations with risk of vitamin A deficiency. Cassava genotypes with high pro-vitamin A activity have been identified as a strategy to reduce the prevalence of deficiency of this vitamin. The data set was used for the construction of a descriptive model by chemometric analysis. The genotypes of yellow-fleshed roots were clustered by the higher concentrations of cis-β-carotene and lutein. Inversely, cream-fleshed roots genotypes were grouped precisely due to their lower concentrations of these pigments, as samples rich in lycopene (red-fleshed) differed among the studied genotypes. The analytical approach (UV-Vis, HPLC, and chemometrics) used showed to be efficient for understanding the chemodiversity of cassava genotypes, allowing to classify them according to important features for human health and nutrition.

UV-visible scanning spectrophotometry and chemometric analysis as tools for carotenoids analysis in cassava genotypes (Manihot esculenta Crantz).

PubMed

Moresco, Rodolfo; Uarrota, Virgílio G; Pereira, Aline; Tomazzoli, Maíra; Nunes, Eduardo da C; Martins Peruch, Luiz Augusto; Gazzola, Jussara; Costa, Christopher; Rocha, Miguel; Maraschin, Marcelo

2015-12-01

In this study, the metabolomics characterization focusing on the carotenoid composition of ten cassava (Manihot esculenta) genotypes cultivated in southern Brazil by UV-visible scanning spectrophotometry and reverse phase-high performance liquid chromatography was performed. Cassava roots rich in β-carotene are an important staple food for populations with risk of vitamin A deficiency. Cassava genotypes with high pro-vitamin A activity have been identified as a strategy to reduce the prevalence of deficiency of this vitamin. The data set was used for the construction of a descriptive model by chemometric analysis. The genotypes of yellow-fleshed roots were clustered by the higher concentrations of cis- β-carotene and lutein. Inversely, cream-fleshed roots genotypes were grouped precisely due to their lower concentrations of these pigments, as samples rich in lycopene (redfleshed) differed among the studied genotypes. The analytical approach (UV-Vis, HPLC, and chemometrics) used showed to be efficient for understanding the chemodiversity of cassava genotypes, allowing to classify them according to important features for human health and nutrition.

Investigating the Effects of Simulated Space conditions on Novel Extremely Halophilic Archaea: Halovarius Luteus gen. nov., sp. nov.

NASA Astrophysics Data System (ADS)

Feshangsaz, Niloofar; Van Loon, ing.. Jack J. W. A.; Nazmi, Kamran; Semsarha, Farid

2016-07-01

Studying halophiles from different environments of Earth provide new insights into our search for life in the universe. Haloarchaea show some unique characteristics and physiological adaptations like acidic proteins against harsh environments such as natural brine with salt concentration approaching saturation (5 M) and regions with low active water. These properties make haloarchaea interesting candidate for astrobiological studies. Halovarius luteus gen. nov., sp. nov. a novel extremely halophilic archaeon from Urmia salt lake, in Iran has been chosen to explore its resistance against a series of extreme conditions. The aim of this study is to assess the resistance of strain DA50T under the effects of simulated space conditions like simulated microgravity, hypergravity, and desiccation. In this paper we will discuss the results of these studies where we specifically focus on changes in carotenoid pigments production and whole cell proteome. This is the first report of very novel Iranian archaea in response to extreme space conditions. The pigments were extracted by acetone and methanol. Pigments were analyzed by scanning the absorbance spectrum in the UV-VIS spectrophotometer. And they were separated by TLC. Whole protein from cell lysate supernatant was extracted after lysis with Bacterial Protein Extraction Reagent and fractionated by RP-HPLC using C18 column. Proteome analyzed by electrophoresis (SDS-PAGE), and MALDI-TOF. Carotenoid pigments are formed under different extreme conditions such as dry environment and gravitational changes. Also the protein composition exhibits alterations after exposure to the same conditions. Our conclusion is that pigments and proteins formation depend on the growth circumstances. Halophiles use this as an adaptation to survive under different environmental conditions.

Correlations Between Macular, Skin, and Serum Carotenoids

PubMed Central

Conrady, Christopher D.; Bell, James P.; Besch, Brian M.; Gorusupudi, Aruna; Farnsworth, Kelliann; Ermakov, Igor; Sharifzadeh, Mohsen; Ermakova, Maia; Gellermann, Werner; Bernstein, Paul S.

2017-01-01

Purpose Ocular and systemic measurement and imaging of the macular carotenoids lutein and zeaxanthin have been employed extensively as potential biomarkers of AMD risk. In this study, we systematically compare dual wavelength retinal autofluorescence imaging (AFI) of macular pigment with skin resonance Raman spectroscopy (RRS) and serum carotenoid levels in a clinic-based population. Methods Eighty-eight patients were recruited from retina and general ophthalmology practices from a tertiary referral center and excluded only if they did not have all three modalities tested, had a diagnosis of macular telangiectasia (MacTel) or Stargardt disease, or had poor AFI image quality. Skin, macular, and serum carotenoid levels were measured by RRS, AFI, and HPLC, respectively. Results Skin RRS measurements and serum zeaxanthin concentrations correlated most strongly with AFI macular pigment volume under the curve (MPVUC) measurements up to 9° eccentricity relative to MPVUC or rotationally averaged macular pigment optical density (MPOD) measurements at smaller eccentricities. These measurements were reproducible and not significantly affected by cataracts. We also found that these techniques could readily identify subjects taking oral carotenoid-containing supplements. Conclusions Larger macular pigment volume AFI and skin RRS measurements are noninvasive, objective, and reliable methods to assess ocular and systemic carotenoid levels. They are an attractive alternative to psychophysical and optical methods that measure MPOD at a limited number of eccentricities. Consequently, skin RRS and MPVUC at 9° are both reasonable biomarkers of macular carotenoid status that could be readily adapted to research and clinical settings. PMID:28728169

[Simultaneous determination of carotenoids and chlorophylls in algae by high performance liquid chromatography].

PubMed

Yuan, J; Zhang, Y; Shi, X; Gong, X; Chen, F

1997-03-01

An isocratic reversed-phase HPLC method for the simultaneous determination of carotenoids and chlorophylls in algae is presented. Both carotenoids and chlorophylls were separated by using a C18 column and a mobile phase consisting of dichloromethane/acetonitrile/methanol/water (22.5:9.5:67.5:0.5). The flow rate was 1.0mL/min. These pigments were detected by a UV-VIS absorbance detector set at 450nm. The average recoveries of these pigments by standard addition method were 99.1% for lutein, 98.5% for alpha-carotene, 99.4% for beta-carotene, 100.6% for chlorophyll a and 99.9% for chlorophyll b. The relative standard deviations of these pigments were 2.4% for lutein, 5.6% for alpha-carotene, 6.0% for beta-carotene, 4.1% for chlorophyll a and 4.0% for chlorophyll b. The calibration curves for these pigments gave good linearity over a wide range of 0-50mg/L (r = 0.9992-0.9999). The retention times were 3.0 min for lutein, 4.7 min for chlorophyll b, 6.3 min for chlorophyll a, 14.1 min for alpha-carotene, and 14.7 min for beta-carotene. Using this method, carotenoids and chlorophylls were determined in Spirulina platensis, Chlorella vulgaris and Haematococcus lacustris. Chlorella vulgaris and Haematococcus lacustris samples were ground in a B. Braun homogenizer. Spirulina platensis sample was smashed by supersonic wave. Acetone was used as extractant.

Anthocyanins and flavonols are responsible for purple color of Lablab purpureus (L.) sweet pods.

PubMed

Cui, Baolu; Hu, Zongli; Zhang, Yanjie; Hu, Jingtao; Yin, Wencheng; Feng, Ye; Xie, Qiaoli; Chen, Guoping

2016-06-01

Lablab pods, as dietary vegetable, have high nutritional values similar to most of edible legumes. Moreover, our studies confirmed that purple lablab pods contain the natural pigments of anthocyanins and flavonols. Compared to green pods, five kinds of anthocyanins (malvidin, delphinidin and petunidin derivatives) were found in purple pods by HPLC-ESI-MS/MS and the major contents were delphinidin derivatives. Besides, nine kinds of polyphenol derivatives (quercetin, myricetin, kaempferol and apigenin derivatives) were detected by UPLC-ESI-MS/MS and the major components were quercetin and myricetin derivatives. In order to discover their molecular mechanism, expression patterns of biosynthesis and regulatory gens of anthocyanins and flavonols were investigated. Experimental results showed that LpPAL, LpF3H, LpF3'H, LpDFR, LpANS and LpPAP1 expressions were significantly induced in purple pods compared to green ones. Meanwhile, transcripts of LpFLS were more abundant in purple pods than green or yellow ones, suggestind that co-pigments of anthocyanins and flavonols are accumulated in purple pods. Under continuously dark condition, no anthocyanin accumulation was detected in purple pods and transcripts of LpCHS, LpANS, LpFLS and LpPAP1 were remarkably repressed, indicating that anthocyanins and flavonols biosynthesis in purple pods was regulated in light-dependent manner. These results indicate that co-pigments of anthocyanins and flavonols contribute to purple pigmentations of pods. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Mycobacterium pseudoshottsii sp. nov., a slowly growing chromogenic species isolated from Chesapeake Bay striped bass (Morone saxatilis)

USGS Publications Warehouse

Rhodes, M.W.; Kator, H.; McNabb, A.; Deshayes, C.; Reyrat, J.-M.; Brown-Elliott, B. A.; Wallace, R.; Trott, K.A.; Parker, J.M.; Lifland, B.; Osterhout, G.; Kaattari, I.; Reece, K.; Vogelbein, W.; Ottinger, C.A.

2005-01-01

A group of slowly growing photochromogenic mycobacteria was isolated from Chesapeake Bay striped bass (Morone saxatilis) during an epizootic of mycobacteriosis. Growth characteristics, acid-fastness and 16S rRNA gene sequencing results were consistent with those of the genus Mycobacterium. Biochemical reactions, growth characteristics and mycolic acid profiles (HPLC) resembled those of Mycobacterium shottsii, a non-pigmented mycobacterium also isolated during the same epizootic. Sequencing of the 16S rRNA genes, the gene encoding the exported repeated protein (erp) and the gene encoding the 65 kDa heat-shock protein (hsp65) and restriction enzyme analysis of the hsp65 gene demonstrated that this group of isolates is unique. Insertion sequences associated with Mycobacterium ulcerans, IS2404 and IS2606, were detected by PCR. These isolates could be differentiated from other slowly growing pigmented mycobacteria by their inability to grow at 37 ??C, production of niacin and urease, absence of nitrate reductase, negative Tween 80 hydrolysis and resistance to isoniazid (1 ??g ml-1), p-nitrobenzoic acid, thiacetazone and thiophene-2-carboxylic hydrazide. On the basis of this polyphasic study, it is proposed that these isolates represent a novel species, Mycobacterium pseudoshottsii sp. nov. The type strain, L15T, has been deposited in the American Type Culture Collection as ATCC BAA-883T and the National Collection of Type Cultures (UK) as NCTC 13318T. ?? 2005 IUMS.

Applications of HPLC/MS in the analysis of traditional Chinese medicines

PubMed Central

Li, Miao; Hou, Xiao-Fang; Zhang, Jie; Wang, Si-Cen; Fu, Qiang; He, Lang-Chong

2012-01-01

In China, traditional Chinese medicines (TCMs) have been used in clinical applications for thousands of years. The successful hyphenation of high-Performance liquid chromatography (HPLC) and mass spectrometry (MS) has been applied widely in TCMs and biological samples analysis. Undoubtedly, HPLC/MS technique has facilitated the understanding of the treatment mechanism of TCMs. We reviewed more than 350 published papers within the last 5 years on HPLC/MS in the analysis of TCMs. The present review focused on the applications of HPLC/MS in the component analysis, metabolites analysis, and pharmacokinetics of TCMs etc. 50% of the literature is related to the component analysis of TCMs, which show that this field is the most populär type of research. In the metabolites analysis, HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry has been demonstrated to be the powerful tool for the characterization of structural features and fragmentation behavior patterns. This paper presented a brief overview of the applications of HPLC/MS in the analysis of TCMs. HPLC/MS in the fingerprint analysis is reviewed elsewhere. PMID:29403684

Carotenoids and Retinoids: Nomenclature, Chemistry, and Analysis.

PubMed

Harrison, Earl H; Curley, Robert W

Carotenoids are polyenes synthesized in plants and certain microorganisms and are pigments used by plants and animals in various physiological processes. Some of the over 600 known carotenoids are capable of metabolic conversion to the essential nutrient vitamin A (retinol) in higher animals. Vitamin A also gives rise to a number of other metabolites which, along with their analogs, are known as retinoids. To facilitate discussion about these important molecules, a nomenclature is required to identify specific substances. The generally accepted rules for naming these important molecules have been agreed to by various Commissions of the International Union of Pure and Applied Chemistry and International Union of Biochemistry. These naming conventions are explained along with comparisons to more systematic naming rules that apply for these organic chemicals. Identification of the carotenoids and retinoids has been advanced by their chemical syntheses, and here, both classical and modern methods for synthesis of these molecules, as well as their analogs, are described. Because of their importance in biological systems, sensitive methods for the detection and quantification of these compounds from various sources have been essential. Early analyses that relied on liquid adsorption and partition chromatography have given way to high-performance liquid chromatography (HPLC) coupled with various detection methods. The development of HPLC coupled to mass spectrometry, particularly LC/MS-MS with Multiple Reaction Monitoring, has resulted in the greatest sensitivity and specificity in these analyses.

Improved analysis of Monascus pigments based on their pH-sensitive UV-Vis absorption and reactivity properties.

PubMed

Shi, Kan; Chen, Gong; Pistolozzi, Marco; Xia, Fenggeng; Wu, Zhenqiang

2016-09-01

Monascus pigments, a mixture of azaphilones mainly composed of red, orange and yellow pigments, are usually prepared in aqueous ethanol and analysed by ultraviolet-visible (UV-Vis) spectroscopy. The pH of aqueous ethanol used during sample preparation and analysis has never been considered a key parameter to control; however, this study shows that the UV-Vis spectra and colour characteristics of the six major pigments are strongly influenced by the pH of the solvent employed. In addition, the increase of solvent pH results in a remarkable increase of the amination reaction of orange pigments with amino compounds, and at higher pH (≥ 6.0) a significant amount of orange pigment derivatives rapidly form. The consequent impact of these pH-sensitive properties on pigment analysis is further discussed. Based on the presented results, we propose that the sample preparation and analysis of Monascus pigments should be uniformly performed at low pH (≤ 2.5) to avoid variations of UV-Vis spectra and the creation of artefacts due to the occurrence of amination reactions, and ensure an accurate analysis that truly reflects pigment characteristics in the samples.

Seed Polyphenolic Profile, Antioxidative Activity, and Fatty Acids Composition of Wild and Cultivated Carthamus Species.

PubMed

Karami, Soraya; Sabzalian, Mohammad R; Rahimmalek, Mehdi

2018-06-01

Total flavonoid content (TFC) and cyanidin-3-glucoside (Cyd-3-glu) of seed and seed coat extract of 16 genotypes from five species of Carthamus were studied, and their major polyphenolic compounds and antioxidant activity of the seed coat extracts were determined using HPLC analysis and DPPH assay, respectively. Additionally, fatty acids composition of the seed oil was analyzed by GC. In general, TFC and Cyd-3-glu content of seed coat extracts were higher than those of seed extracts. A novel breeding line with black seed coat (named A82) depicted lower TFC (3.79 mg QUE/g DW) but higher Cyd-3-glu (24.64 mg/g DW) compared to the white and other seed-pigmented genotypes. DPPH radical scavenging activity showed a strong association with Cyd-3-glu content (r = 0.84), but no correlation with TFC (r = -0.32). HPLC analysis of seed coat extracts revealed that four compounds were dominant constituents including rutin (7.23 - 117.95 mg/100 g DW), apigenin (4.37 - 64.88 mg/100 g DW), quercetin (3.09 - 14.10 mg/100 g DW), and ferulic acid (4.49 - 30.41 mg/100 g DW). Interestingly, the genotype A82 with an appropriate polyunsaturated/saturated fatty acids index (5.46%) and a moderate linoleic fatty acid content (64.70%) had higher nutritional and pharmaceutical value than all the other genotypes. © 2018 Wiley-VHCA AG, Zurich, Switzerland.

Terahertz Analysis of Quinacridone Pigments

NASA Astrophysics Data System (ADS)

Squires, A. D.; Kelly, M.; Lewis, R. A.

2017-03-01

We present terahertz spectroscopy and analysis of two commercially available quinacridone pigments in the 0.5-4.5 THz range. Our results show a clear distinction between quinacridone red and magenta pigments. We reveal four definite absorptions in the terahertz regime common to both pigments, but offset between the pigments by ˜0.2 THz. The lowest-energy line in each pigment is observed to increase in frequency by ˜0.1 THz as the temperature is reduced from 300 to 12 K.

Isolation and purification of all-trans diadinoxanthin and all-trans diatoxanthin from diatom Phaeodactylum tricornutum.

PubMed

Kuczynska, Paulina; Jemiola-Rzeminska, Malgorzata

2017-01-01

Two diatom-specific carotenoids are engaged in the diadinoxanthin cycle, an important mechanism which protects these organisms against photoinhibition caused by absorption of excessive light energy. A high-performance and economical procedure of isolation and purification of diadinoxanthin and diatoxanthin from the marine diatom Phaeodactylum tricornutum using a four-step procedure has been developed. It is based on the use of commonly available materials and does not require advanced technology. Extraction of pigments, saponification, separation by partition and then open column chromatography, which comprise the complete experimental procedure, can be performed within 2 days. This method allows HPLC grade diadinoxanthin and diatoxanthin of a purity of 99 % or more to be obtained, and the efficiency was estimated to be 63 % for diadinoxanthin and 73 % for diatoxanthin. Carefully selected diatom culture conditions as well as analytical ones ensure highly reproducible performance. A protocol can be used to isolate and purify the diadinoxanthin cycle pigments both on analytical and preparative scale.

Structure of the higher plant light harvesting complex I: in vivo characterization and structural interdependence of the Lhca proteins.

PubMed

Klimmek, Frank; Ganeteg, Ulrika; Ihalainen, Janne A; van Roon, Henny; Jensen, Poul E; Scheller, Henrik V; Dekker, Jan P; Jansson, Stefan

2005-03-01

We have investigated the structure of the higher plant light harvesting complex of photosystem I (LHCI) by analyzing PSI-LHCI particles isolated from a set of Arabidopsis plant lines, each lacking a specific Lhca (Lhca1-4) polypeptide. Functional antenna size measurements support the recent finding that there are four Lhca proteins per PSI in the crystal structure [Ben-Shem, A., Frolow, F., and Nelson, N. (2003) Nature 426, 630-635]. According to HPLC analyses the number of pigment molecules bound within the LHCI is higher than expected from reconstitution studies or analyses of isolated native LHCI. Comparison of the spectra of the particles from the different lines reveals chlorophyll absorption bands peaking at 696, 688, 665, and 655 nm that are not present in isolated PSI or LHCI. These bands presumably originate from "gap" or "linker" pigments that are cooperatively coordinated by the Lhca and/or PSI proteins, which we have tentatively localized in the PSI-LHCI complex.

Content of phenolic compounds and free polyamines in black chokeberry (Aronia melanocarpa) after application of polyamine biosynthesis regulators.

PubMed

Hudec, Jozef; Bakos, Dusan; Mravec, Dusan; Kobida, L'ubomír; Burdová, Maria; Turianica, Ivan; Hlusek, Jaroslav

2006-05-17

The total contents of anthocyanins, flavonoids, and phenolics in 60 samples of black chokeberries (Aronia melanocarpa), after treating with catabolites of polyamine biosynthesis (KPAb) and ornithine decarboxylase inhibitor, were analyzed spectrophotometrically, and quercetin and free polyamine contents were analyzed by RP-HPLC with UV detection. The average total contents of the individual substances and phenolic subgroups in control berries were as follows (mg x kg(-1)): anthocyanines, 6408; flavonoids, 664; phenolics, 37,600; quercetin, 349. KPAb decreased total contents of anthocyanines and phenolics only slightly but significantly increased the content of flavonoids. This caused an important change in the abundance of flavonoids in the pigment complex. The absolute content of quercetin was increased, but its ratio to flavonoids content was decreased. Ornithine decarboxylase inhibitor had a markedly different effect as it significantly increased total content of anthocyanins and total phenolics, inhibited the total content of free polyamines, and stimulated the processes of saccharides transformation to phenolic pigments.

Optimal methodologies for terahertz time-domain spectroscopic analysis of traditional pigments in powder form

NASA Astrophysics Data System (ADS)

Ha, Taewoo; Lee, Howon; Sim, Kyung Ik; Kim, Jonghyeon; Jo, Young Chan; Kim, Jae Hoon; Baek, Na Yeon; Kang, Dai-ill; Lee, Han Hyoung

2017-05-01

We have established optimal methods for terahertz time-domain spectroscopic analysis of highly absorbing pigments in powder form based on our investigation of representative traditional Chinese pigments, such as azurite [blue-based color pigment], Chinese vermilion [red-based color pigment], and arsenic yellow [yellow-based color pigment]. To accurately extract the optical constants in the terahertz region of 0.1 - 3 THz, we carried out transmission measurements in such a way that intense absorption peaks did not completely suppress the transmission level. This required preparation of pellet samples with optimized thicknesses and material densities. In some cases, mixing the pigments with polyethylene powder was required to minimize absorption due to certain peak features. The resulting distortion-free terahertz spectra of the investigated set of pigment species exhibited well-defined unique spectral fingerprints. Our study will be useful to future efforts to establish non-destructive analysis methods of traditional pigments, to construct their spectral databases, and to apply these tools to restoration of cultural heritage materials.

A Rapid Method for the Determination of Fucoxanthin in Diatom

PubMed Central

Wang, Li-Juan; Fan, Yong; Parsons, Ronald L.; Hu, Guang-Rong; Zhang, Pei-Yu

2018-01-01

Fucoxanthin is a natural pigment found in microalgae, especially diatoms and Chrysophyta. Recently, it has been shown to have anti-inflammatory, anti-tumor, and anti-obesityactivity in humans. Phaeodactylum tricornutum is a diatom with high economic potential due to its high content of fucoxanthin and eicosapentaenoic acid. In order to improve fucoxanthin production, physical and chemical mutagenesis could be applied to generate mutants. An accurate and rapid method to assess the fucoxanthin content is a prerequisite for a high-throughput screen of mutants. In this work, the content of fucoxanthin in P. tricornutum was determined using spectrophotometry instead of high performance liquid chromatography (HPLC). This spectrophotometric method is easier and faster than liquid chromatography and the standard error was less than 5% when compared to the HPLC results. Also, this method can be applied to other diatoms, with standard errors of 3–14.6%. It provides a high throughput screening method for microalgae strains producing fucoxanthin. PMID:29361768

A simple method for plasma total vitamin C analysis suitable for routine clinical laboratory use.

PubMed

Robitaille, Line; Hoffer, L John

2016-04-21

In-hospital hypovitaminosis C is highly prevalent but almost completely unrecognized. Medical awareness of this potentially important disorder is hindered by the inability of most hospital laboratories to determine plasma vitamin C concentrations. The availability of a simple, reliable method for analyzing plasma vitamin C could increase opportunities for routine plasma vitamin C analysis in clinical medicine. Plasma vitamin C can be analyzed by high performance liquid chromatography (HPLC) with electrochemical (EC) or ultraviolet (UV) light detection. We modified existing UV-HPLC methods for plasma total vitamin C analysis (the sum of ascorbic and dehydroascorbic acid) to develop a simple, constant-low-pH sample reduction procedure followed by isocratic reverse-phase HPLC separation using a purely aqueous low-pH non-buffered mobile phase. Although EC-HPLC is widely recommended over UV-HPLC for plasma total vitamin C analysis, the two methods have never been directly compared. We formally compared the simplified UV-HPLC method with EC-HPLC in 80 consecutive clinical samples. The simplified UV-HPLC method was less expensive, easier to set up, required fewer reagents and no pH adjustments, and demonstrated greater sample stability than many existing methods for plasma vitamin C analysis. When compared with the gold-standard EC-HPLC method in 80 consecutive clinical samples exhibiting a wide range of plasma vitamin C concentrations, it performed equivalently. The easy set up, simplicity and sensitivity of the plasma vitamin C analysis method described here could make it practical in a normally equipped hospital laboratory. Unlike any prior UV-HPLC method for plasma total vitamin C analysis, it was rigorously compared with the gold-standard EC-HPLC method and performed equivalently. Adoption of this method could increase the availability of plasma vitamin C analysis in clinical medicine.

Antioxidant activities of Vaccinium uliginosum L. extract and its active components.

PubMed

Kim, Young-Hee; Bang, Chae-Young; Won, Eun-Kyung; Kim, Jong-Pyung; Choung, Se-Young

2009-08-01

Vaccinium uliginosum L. (also known as bog bilberry) is a low-growing deciduous shrub classified in the Ericaceae family of plants, which includes numerous Vaccinium berries, blueberries, and cranberries. Berries of the Ericaceae family are known to contain organic acids, vitamins, glycosides, and anthocyanins and have been reported to have antioxidant activity. In order to identify the antioxidative principles of V. uliginosum, we separated water extracts into polyphenol, anthocyanin-rich (pigment), and sugar/acid fractions by using ethyl acetate, acidic methanol (MeOH), and 0.01 N HCl. Antioxidant activities were assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH), superoxide radical, and hydroxyl radical assays. The crude extract and fractions containing polyphenol and pigment exhibited the greatest antioxidant activities with 50% inhibitory concentration (IC(50)) values of 85.8 microg/mL, 33.2 microg/mL, and 16.7 microg/mL, respectively, for the DPPH assay and 48.1 microg/mL, 83.8 microg/mL, and 51.9 microg/mL for the nonenzymatic superoxide radical assay. The fractions containing polyphenol, pigment, and sugar/acid significantly inhibited xanthine oxidase. To investigate the functional compounds from the active fractions, we purified the polyphenol fraction and separated the compounds by using chromatographic techniques. The crude extract was dissolved in MeOH and further purified by reversed-phase high-performance liquid chromatography (HPLC) using MeOH-water (35:65 vol/vol) (with 0.04% trifluoroacetic acid) to obtain VU-EA-1 (16.6 mg), VU-EA-2 (8.5 mg), VU-EA-3 (19.8 mg), VU-EA-4 (12.8 mg), VU-EA-5 (6.5 mg), and VU-EA-6 (23.5 mg). The MeOH-washed fraction from the HPLC was concentrated and purified by reversed-phase HPLC using MeOH-water (50:50 vol/vol) to give VU-EA-10 (12.4 mg). Antioxidant activity was assessed by DPPH, superoxide radical, and hydroxyl radical assays. The isolated compounds exhibited dose-dependent antioxidant activity with IC(50) values of 7.6 microg/mL (VU-EA-10) for the DPPH assay, 67.8 microg/mL (VU-EA-4) for the nonenzymatic superoxide radical assay, and 3.7 microg/mL (VU-EA-10) and 7.6 microg/ml (VU-EA-6) for the enzymatic superoxide radical assay and 30% inhibitory concentration values of 0.58 microg/mL (VU-EA-1), 0.57 microg/mL (VU-EA-5), and 0.70 microg/mL (VU-EA-6) for the hydroxyl radical assay. In conclusion, V. uliginosum had potent antioxidative activity, and flavonoids were isolated as the main active principles.

Cluster analysis of historical and modern hard red spring wheat cultivars based on parentage and HPLC analysis of gluten forming proteins

USDA-ARS?s Scientific Manuscript database

In this study, 30 hard red spring (HRS) wheat cultivars released between 1910 and 2013 were analyzed to determine how they cluster in terms of parentage and protein data, analyzed by reverse-phase HPLC (RP-HPLC) of gliadins, and size-exclusion HPLC (SE-HPLC) of unreduced proteins. Dwarfing genes in...

Carotenoid stability during production and storage of tomato juice made from tomatoes with diverse pigment profiles measured by infrared spectroscopy.

PubMed

Rubio-Diaz, Daniel E; Santos, Alejandra; Francis, David M; Rodriguez-Saona, Luis E

2010-08-11

Chemical changes in carotenoids and lipids were studied during production and storage of canned tomato juice using ATR infrared spectroscopy and HPLC. Samples from 10 groups of tomatoes with different carotenoid profiles were analyzed fresh, after hot-break and screening, after canning, and at five different time points during 1 year of storage. An apparent increase of carotenoids was observed after hot-break due to improved extraction efficiency. This increase was accompanied by some degree of lipid oxidation and carotenoid isomerization. Canning produced the most intense changes in the lipid profile with breakdown of triglycerides ( approximately 1743 cm(-1)), formation of fatty acids ( approximately 1712 cm(-1)), and degradation and isomerization of trans-carotenoids ( approximately 960 and approximately 3006 cm(-1)). Isomerization was corroborated by the relative increase of HPLC areas corresponding to carotenoid cis isomers. Canning reduced trans-lycopene, trans-delta-carotene, trans-beta-carotene, and trans-lutein by 30, 34, 43, and 67%, respectively. HPLC data indicate that canning causes a drastic reduction of tetra-cis-lycopene and promotes its isomerization to other geometric forms, including all-trans-lycopene. Infrared spectra of tomato juice lipid fractions correlated well with the number of days in storage (SECV < 11 days, r values > 0.99), demonstrating continuous degradation of lipids. Results demonstrated that individual carotenoids and their isomeric forms behave differently during production and storage of canned tomato juice. Information collected by infrared spectroscopy complemented well that of HPLC, providing marker bands to further the understanding of chemical changes taking place during processing and storage of tomato juice.

Colorimetric Analysis of Hibiscus Beverages and their Potential Antioxidant Properties.

PubMed

Camelo-Méndez, G A; Vanegas-Espinoza, P E; Escudero-Gilete, M L; Heredia, F J; Paredes-López, O; Del Villar-Martínez, A A

2018-05-25

In food industry, roselle beverages and their subproducts could be functional ingredients since they are an excellent source of bioactive compounds with improved performance due to their important anthocyanins content. The aim of this study was to analyze anthocyanin content and antioxidant properties of aqueous infusions elaborated with color contrasting Hibiscus materials and design a mathematical model in order to predict color-composition relationship. Color measurements of beverages from roselle (Negra, Sudan and Rosa) were made by transmission spectrophotometry, anthocyanins quantification was determined by HPLC, and antioxidant potential was evaluated by in vitro methods (ABTS and FRAP assays). Beverages prepared with particle size minor of 250 μm presented until 4- and 2- times more anthocyanins content and antioxidant capacity respectively, in comparison to beverages prepared with powders with particle size major of 750 μm. Positive correlations among pigments composition and color parameters were found (p < 0.05), showing that anthocyanins content, antioxidant capacity, C* ab and h ab values increased in relation with the smallest particle size of flours. Also, mathematical models were stablished to predict anthocyanin content (r ≥ 0.97) and antioxidant capacity (r ≥ 0.89) from color data; we propose equations for quick estimation of the antioxidant capacity in the Hibiscus beverages with high anthocyanin content. The obtained models could be an important tool to be used in food industry for pigment characterization or functional compounds with potential health benefits.

Internal and external factors affecting photosynthetic pigment composition in plants: a meta-analytical approach.

PubMed

Esteban, Raquel; Barrutia, Oihana; Artetxe, Unai; Fernández-Marín, Beatriz; Hernández, Antonio; García-Plazaola, José Ignacio

2015-04-01

Photosynthetic pigment composition has been a major study target in plant ecophysiology during the last three decades. Although more than 2000 papers have been published, a comprehensive evaluation of the responses of photosynthetic pigment composition to environmental conditions is not yet available. After an extensive survey, we compiled data from 525 papers including 809 species (subkingdom Viridiplantae) in which pigment composition was described. A meta-analysis was then conducted to assess the ranges of photosynthetic pigment content. Calculated frequency distributions of pigments were compared with those expected from the theoretical pigment composition. Responses to environmental factors were also analysed. The results revealed that lutein and xanthophyll cycle pigments (VAZ) were highly responsive to the environment, emphasizing the high phenotypic plasticity of VAZ, whereas neoxanthin was very stable. The present meta-analysis supports the existence of relatively narrow limits for pigment ratios and also supports the presence of a pool of free 'unbound' VAZ. Results from this study provide highly reliable ranges of photosynthetic pigment contents as a framework for future research on plant pigments. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

Effects of glycolic acid chemical peeling on facial pigment deposition: evaluation using novel computer analysis of digital-camera-captured images.

PubMed

Kakudo, Natsuko; Kushida, Satoshi; Suzuki, Kenji; Kusumoto, Kenji

2013-12-01

Chemical peeling is becoming increasingly popular for skin rejuvenation in dermatological cosmetic medicine. However, the improvements seen with chemical peeling are often very minor, and it is difficult to conduct a quantitative assessment of pre- and post-treatment appearance. We report the pre- and postpeeling effects for facial pigment deposition using a novel computer analysis method for digital-camera-captured images. Glycolic acid chemical peeling was performed a total of 5 times at 2-week intervals in 23 healthy women. We conducted a computer image analysis by utilizing Robo Skin Analyzer CS 50 and Clinical Suite 2.1 and then reviewed each parameter for the area of facial pigment deposition pre- and post-treatment. Parameters were pigmentation size and four pigmentation categories: little pigmentation and three levels of marked pigmentation (Lv1, 2, and 3) based on detection threshold. Each parameter was measured, and the total area of facial pigmentation was calculated. The total area of little pigmentation and marked pigmentation (Lv1) was significantly reduced. On the other hand, a significant difference was not observed for the total area of marked pigmentation Lv2 and Lv3. This suggests that glycolic acid chemical peeling has an effect on small facial pigment disposition or has an effect on light pigment deposition. As the Robo Skin Analyzer is useful for objectively quantifying and analyzing minor changes in facial skin, it is considered to be an effective tool for accumulating treatment evidence in the cosmetic and esthetic skin field. © 2013 Wiley Periodicals, Inc.

Retinal accumulation of zeaxanthin, lutein, and β-carotene in mice deficient in carotenoid cleavage enzymes.

PubMed

Li, Binxing; Vachali, Preejith P; Shen, Zhengqing; Gorusupudi, Aruna; Nelson, Kelly; Besch, Brian M; Bartschi, Alexis; Longo, Simone; Mattinson, Ty; Shihab, Saeed; Polyakov, Nikolay E; Suntsova, Lyubov P; Dushkin, Alexander V; Bernstein, Paul S

2017-06-01

Carotenoid supplementation can prevent and reduce the risk of age-related macular degeneration (AMD) and other ocular disease, but until now, there has been no validated and well-characterized mouse model which can be employed to investigate the protective mechanism and relevant metabolism of retinal carotenoids. β-Carotene oxygenases 1 and 2 (BCO1 and BCO2) are the only two carotenoid cleavage enzymes found in animals. Mutations of the bco2 gene may cause accumulation of xanthophyll carotenoids in animal tissues, and BCO1 is involved in regulation of the intestinal absorption of carotenoids. To determine whether or not mice deficient in BCO1 and/or BCO2 can serve as a macular pigment mouse model, we investigated the retinal accumulation of carotenoids in these mice when fed with zeaxanthin, lutein, or β-carotene using an optimized carotenoid feeding method. HPLC analysis revealed that all three carotenoids were detected in sera, livers, retinal pigment epithelium (RPE)/choroids, and retinas of all of the mice, except that no carotenoid was detectable in the retinas of wild type (WT) mice. Significantly higher amounts of zeaxanthin and lutein accumulated in the retinas of BCO2 knockout (bco2 -/- ) mice and BCO1/BCO2 double knockout (bco1 -/- /bco2 -/- ) mice relative to BCO1 knockout (bco1 -/- ) mice, while bco1 -/- mice preferred to take up β-carotene. The levels of zeaxanthin and lutein were higher than β-carotene levels in the bco1 -/- /bco2 -/- retina, consistent with preferential uptake of xanthophyll carotenoids by retina. Oxidative metabolites were detected in mice fed with lutein or zeaxanthin but not in mice fed with β-carotene. These results indicate that bco2 -/- and bco1 -/- /bco2 -/- mice could serve as reasonable non-primate models for macular pigment function in the vertebrate eye, while bco1 -/- mice may be more useful for studies related to β-carotene. Copyright © 2017 Elsevier Ltd. All rights reserved.

Size-dependent depletion and community disturbance of phytoplankton under intensive oyster mariculture based on HPLC pigment analysis in Daya Bay, South China Sea.

PubMed

Jiang, Tao; Chen, Feiyu; Yu, Zonghe; Lu, Lin; Wang, Zhaohui

2016-12-01

In this study, we conducted a 14-month investigation in Daya Bay, southern China to understand the effects of oyster farming on phytoplankton community and biomass by using size-fractionated phytopigments. Results proved the filtering effects of oysters on phytoplankton biomass. During the oyster culture period, the average concentration of total chlorophyll a (sum of size-fractionated Chl a) within the farming area was approximately 60% lower than that at the reference site. Phytoplankton depletion in the aquaculture zone mainly occurred in micro-sized fractions (>20 μm) of Chl a, fucoxanthin, and peridinin. The influence of oyster filtration on nano-sized (2.7-20 μm) pigments seemed less than that on micro-sized ones. The depletion of peridinin and 19'-hex-fucoxanthin in aquaculture zone was higher than those of the other pigments, which indicated that flagellated cells might be selectively filtered by oysters and could be more easily influenced by oyster aquaculture. The pico-sized Chl a (<2.7 μm) comprised 24% of total Chl a on the average in the aquaculture zone during the cultural period compared to 6% in the reference site. Picoeukaryote abundance, which was determined via flow cytometry, was significantly higher in the aquaculture zone than in the non-aquaculture areas. The abundance of picoeukaryote is significantly and positively correlated with the concentrations of pico-sized prasinoxanthin, violaxanthin, and neoxanthin, indicating that picoeukaryote is dominated by those in prasinophyte. The results suggest that oyster aquaculture might stimulate the growth of prasinophyte, although the seasonal variations are mainly controlled by the water temperature in the study area. This research highlights the successful use of size-fractionated phytopigments to estimate size-specific phytoplankton biomass and community, which can be applied as a routine method to monitor the environmental effect and food resources of bivalve aquaculture. Copyright © 2016 Elsevier Ltd. All rights reserved.

Free radical scavenging and anti-oxidative activities of an ethanol-soluble pigment extract prepared from fermented Zijuan Pu-erh tea.

PubMed

Fan, Jiang Ping; Fan, Chong; Dong, Wen Min; Gao, Bin; Yuan, Wei; Gong, Jia Shun

2013-09-01

An ethanol-soluble pigment extract was separated from fermented Zijuan Pu-erh tea. The compositions of the ethanol soluble pigment extract were analyzed by high-performance liquid chromatography-tandem mass spectroscopy (HPLC-MS/MS). The extract was prepared into a series of ethanol solutions and analyzed for free radical-scavenging activities (against two free radicals: 1,1-diphenyl-2-picrylhydrazyl (DPPH) and (2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO)) and in vitro anti-oxidative properties. Electron spin resonance spectroscopy showed that the peaks of DPPH and TEMPO decreased with increasing extract concentration, suggesting that the extract had excellent free radical-scavenging activities. In vitro cell culture suggested that, at 50-200 mg/L, the extract had no measurable effect on the viability of vascular endothelial cells (ECV340) but produced significant protective effects for cells that underwent oxidative injuries due to hydrogen peroxide (H₂O₂) treatment. Compared with the H₂O₂ treatment alone cells group, 200 mg/L of the extract increased the activity of superoxide dismutase (SOD) in cells by 397.3%, and decreased the concentration of malondialdehyde (MDA) and the activity of lactate acid dehydrogenase (LDH) by 47.8% and 69.6%, respectively. These results suggest that the extract has excellent free radical scavenging and anti-oxidative properties. Copyright © 2013 Elsevier Ltd. All rights reserved.

Self-Normalized Photoacoustic Technique for the Quantitative Analysis of Paper Pigments

NASA Astrophysics Data System (ADS)

Balderas-López, J. A.; Gómez y Gómez, Y. M.; Bautista-Ramírez, M. E.; Pescador-Rojas, J. A.; Martínez-Pérez, L.; Lomelí-Mejía, P. A.

2018-03-01

A self-normalized photoacoustic technique was applied for quantitative analysis of pigments embedded in solids. Paper samples (filter paper, Whatman No. 1), attached with the pigment: Direct Fast Turquoise Blue GL, were used for this study. This pigment is a blue dye commonly used in industry to dye paper and other fabrics. The optical absorption coefficient, at a wavelength of 660 nm, was measured for this pigment at various concentrations in the paper substrate. It was shown that Beer-Lambert model for light absorption applies well for pigments in solid substrates and optical absorption coefficients as large as 220 cm^{-1} can be measured with this photoacoustic technique.

Development of green extraction processes for Nannochloropsis gaditana biomass valorization.

PubMed

Sánchez-Camargo, Andrea Del Pilar; Pleite, Natalia; Mendiola, José Antonio; Cifuentes, Alejandro; Herrero, Miguel; Gilbert-López, Bienvenida; Ibáñez, Elena

2018-04-23

In the present work, the valorization of Nannochloropsis gaditana biomass is proposed within the concept of biorefinery. To this aim, high-pressure homogenization (HPH) was used to break down the strong cell wall and supercritical fluid extraction (SFE) with pure CO 2 was applied as a first step to extract valuable compounds (such as non-polar lipids and pigments). Extraction of the remaining residue for the recovery of bioactive compounds was studied by means of an experimental design based on response surface methodology (RSM) employing pressurized liquid extraction (PLE) with green solvents such as water and ethanol. Optimum extract was achieved with pure ethanol at 170°C for 20 min, providing an important antioxidant capacity (0.72 ± 0.03 mmol trolox eq g -1 extract). Complete chemical characterization of the optimum extract was carried out by using different chromatographic methods such as reverse-phase high-performance liquid chromatography with diode array detection (RP-HPLC-DAD), normal-phase HPLC with evaporative light scattering detection (NP-HPLC-ELSD) and gas chromatography coupled to mass spectrometry detection (GC-MS); carotenoids (e.g. violaxanthin), chlorophylls and polar lipids were the main compounds observed while palmitoleic, palmitic, myristic acids and the polyunsaturated eicosapentanoic (EPA) acid were the predominant fatty acids in all PLE extracts. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The identification of synthetic organic pigments in modern paints and modern paintings using pyrolysis-gas chromatography-mass spectrometry.

PubMed

Russell, Joanna; Singer, Brian W; Perry, Justin J; Bacon, Anne

2011-05-01

A collection of more than 70 synthetic organic pigments were analysed using pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS). We report on the analysis of diketo-pyrrolo-pyrrole, isoindolinone and perylene pigments which are classes not previously reported as being analysed by this technique. We also report on a number of azo pigments (2-naphthol, naphthol AS, arylide, diarylide, benzimidazolone and disazo condensation pigments) and phthalocyanine pigments, the Py-GC-MS analysis of which has not been previously reported. The members of each class were found to fragment in a consistent way and the pyrolysis products are reported. The technique was successfully applied to the analysis of paints used by the artist Francis Bacon (1909-1992), to simultaneously identify synthetic organic pigments and synthetic binding media in two samples of paint taken from Bacon's studio and micro-samples taken from three of his paintings and one painting attributed to him.

Profiling and quantification of grain anthocyanins in purple pericarp × blue aleurone wheat crosses by high-performance thin-layer chromatography and densitometry.

PubMed

Böhmdorfer, Stefan; Oberlerchner, Josua Timotheus; Fuchs, Christina; Rosenau, Thomas; Grausgruber, Heinrich

2018-01-01

Anthocyanins are abundant secondary metabolites responsible for most blue to blue-black, and red to purple colors of various plant organs. In wheat grains, anthocyanins are accumulated in the pericarp and/or aleurone layer. Anthocyanin pigmented wheat grains can be processed into functional foods with potential health benefits due to the antioxidant properties of the anthocyanins. The grain anthocyanin content can be increased by pyramidizing the different genes responsible for the accumulation of anthocyanins in the different grain layers. Our objective was to develop a high-performance thin-layer chromatography (HPTLC) method that allows the determination of both the anthocyanin profile and the total pigment concentration. Thereby, selection of breeding lines with significantly higher grain anthocyanin content from purple pericarp × blue aleurone wheat crosses should become more efficient than selection based on only visual scoring of grain color and the unspecific determination of anthocyanin concentration by UV/Vis spectroscopy. A wide variability in the grain anthocyanin content was observed in breeding lines and check varieties. The highest concentration of anthocyanins was observed in deep purple (i.e. combination of the purple pericarp and blue aleurone genetics) grained breeding lines, followed by blue aleurone and purple pericarp genotypes. Determination of the total anthocyanin content was included into the chromatographic analysis, rendering an additional photometric analysis unnecessary. Ten target zones were identified in anthocyanin pigmented wheat grains; four of these zones were typically for blue aleurone types, five for purple pericarp types, and one (i.e. kuromanin glucoside) was characteristic for both. Chemometrics applied to the anthocyanin profile recorded by scanning densitometry revealed that peak heights and peak areas are highly correlated and that seven out of the ten target zones were responsible for about 90% of the total variation in the germplasm. Multivariate analysis of these seven target zones allowed not only a separation of the genetic material into purple, blue and deep purple grained genotypes, but also the identification of genotypes with a specific anthocyanin pattern. Thereby, the original classification by visual scoring was overruled in about one-third of the breeding lines. The presented HPTLC method with à côté calibration allowed the profiling of the pigments and quantification of wheat grain anthocyanin content in a single analysis, replacing UV/Vis spectroscopy with subsequent HPLC analysis. Moreover, no sample preparation apart from extraction and filtration is required, and more than 15 samples can be evaluated in one analysis run, corresponding to several dozens of samples per day. Hence, the method fulfills the requirements for screening methods in early generations of a plant breeding program such as high-throughput, small sample size, high repeatability, fast determination, and reasonable costs per sample. Combined with multivariate statistical analysis, the anthocyanin pattern allowed the validation of the genetic background in the offspring of purple × blue wheat crosses and, therefore, the efficient selection of genotypes exhibiting both the cyanidin and delphinidin aglycon.

[Analysis of pigments from Rhodotorula glutinis by Raman spectroscopy and thin layer chromatography].

PubMed

Yuan, Yu-feng; Tao, Zhan-hua; Wang, Xue; Li, Yong-qing; Liu, Jun-xian

2012-03-01

The pigments from Rhodotorula glutinis were separated by using thin layer chromatography, and the result showed that Rhodotorula glutinis cells could synthesize at least three kinds of pigments, which were beta-carotene, torulene, and torularhodin. The Raman spectra based on the three pigments were acquired, and original spectra were preprocessed by background elimination, baseline correction, and three-point-smoothing, then the averaged spectra from different pigments were investigated, and the result indicated that Raman shift which represents C-C bond was different, and the wave number of beta-carotene demonstrated the largest deviation, finally torulene and torularhodin in Rhodotorula glutinis had more content than beta-carotene. Quantitative analysis of Raman peak height ratio revealed that peak height ratio of pigments showed little difference, which could be used as parameters for further research on living cells, providing reference content of pigments. The above results suggest that Raman spectroscopy combined with thin layer chromatography can be applied to analyze pigments from Rhodotorula glutinis, provides abundant information about pigments, and serves as an effective method to study pigments.

Vertical distribution of phytoplankton communities in open ocean: An assessment based on surface chlorophyll

NASA Astrophysics Data System (ADS)

Uitz, Julia; Claustre, Hervé; Morel, André; Hooker, Stanford B.

2006-08-01

The present study examines the potential of using the near-surface chlorophyll a concentration ([Chla]surf), as it can be derived from ocean color observation, to infer the column-integrated phytoplankton biomass, its vertical distribution, and ultimately the community composition. Within this context, a large High-Performance Liquid Chromatography (HPLC) pigment database was analyzed. It includes 2419 vertical pigment profiles, sampled in case 1 waters with various trophic states (0.03-6 mg Chla m-3). The relationships between [Chla]surf and the chlorophyll a vertical distribution, as previously derived by Morel and Berthon (1989), are fully confirmed. This agreement makes it possible to go further and to examine if similar relationships between [Chla]surf and the phytoplankton assemblage composition along the vertical can be derived. Thanks to the detailed pigment composition, and use of specific pigment biomarkers, the contribution to the local chlorophyll a concentration of three phytoplankton groups can be assessed. With some cautions, these groups coincide with three size classes, i.e., microplankton, nanoplankton and picoplankton. Corroborating previous regional findings (e.g., large species dominate in eutrophic environments, whereas tiny phytoplankton prevail in oligotrophic zones), the present results lead to an empirical parameterization applicable to most oceanic waters. The predictive skill of this parameterization is satisfactorily tested on a separate data set. With such a tool, the vertical chlorophyll a profiles of each group can be inferred solely from the knowledge of [Chla]surf. By combining this tool with satellite ocean color data, it becomes possible to quantify on a global scale the phytoplankton biomass associated with each of the three algal assemblages.

Anthocyanin and Carotenoid Contents in Different Cultivars of Chrysanthemum (Dendranthema grandiflorum Ramat.) Flower.

PubMed

Park, Chang Ha; Chae, Soo Cheon; Park, Soo-Yun; Kim, Jae Kwang; Kim, Yong Joo; Chung, Sun Ok; Arasu, Mariadhas Valan; Al-Dhabi, Naif Abdullah; Park, Sang Un

2015-06-15

The flowers of twenty-three cultivars of Dendranthema grandiflorum Ramat. were investigated to determine anthocyanin and carotenoid levels and to confirm the effects of the pigments on the flower colors using high-performance liquid chromatography (HPLC) and electrospray ionization-mass spectrometry (ESI-MS). The cultivars contained the anthocyanins cyanidin 3-glucoside (C3g) and cyanidin 3-(3"-malonoyl) glucoside (C3mg) and the following carotenoids: lutein, zeaxanthin, β-cryptoxanthin, 13-cis-β-carotene, α-carotene, trans-β-carotene, and 9-cis-β-carotene. The cultivar "Magic" showed the greatest accumulation of total and individual anthocyanins, including C3g and C3gm. On the other hand, the highest level of lutein and zeaxanthin was noted in the cultivar "Il Weol". The cultivar "Anastasia" contained the highest amount of carotenoids such as trans-β-carotene, 9-cis-β-carotene, and 13-cis-β-carotene. The highest accumulation of β-cryptoxanthin and α-carotene was noted in the cultivar "Anastasia" and "Il Weol". Our results suggested that 'Magic", "Angel" and "Relance' had high amounts of anthocyanins and showed a wide range of red and purple colors in their petals, whereas "Il Weol', "Popcorn Ball' and "Anastasia" produced higher carotenoid contents and displayed yellow or green petal colors. Interestingly, "Green Pang Pang", which contained a high level of anthocyanins and a medium level of carotenoids, showed the deep green colored petals. "Kastelli", had high level of carotenoids as well as a medium level of anthocyanins and showed orange and red colored petals. It was concluded that each pigment is responsible for the petal's colors and the compositions of the pigments affect their flower colors and that the cultivars could be a good source for pharmaceutical, floriculture, and pigment industries.

Hyperspectral analysis of cultural heritage artifacts: pigment material diversity in the Gough Map of Britain

NASA Astrophysics Data System (ADS)

Bai, Di; Messinger, David W.; Howell, David

2017-08-01

The Gough Map, one of the earliest surviving maps of Britain, was created and extensively revised over the 15th century. In 2015, the map was imaged using a hyperspectral imaging system while in the collection at the Bodleian Library, Oxford University. The goal of the collection of the hyperspectral image (HSI) of the Gough Map was to address questions such as enhancement of faded text for reading and analysis of the pigments used during its creation and revision. In particular, pigment analysis of the Gough Map will help historians understand the material diversity of its composition and potentially the timeline of, and methods used in, the creation and revision of the map. Multiple analysis methods are presented to analyze a particular pigment in the Gough Map with an emphasis on understanding the within-material diversity, i.e., the number and spatial layout of distinct red pigments. One approach for understanding the number of distinct materials in a scene (i.e., endmember selection and dimensionality estimation) is the Gram matrix approach. Here, this method is used to study the within-material differences of pigments in the map with common visual color. The application is a pigment analysis tool that extracts visually common pixels (here, the red pigments) from the Gough Map and estimates the material diversity of the pixels. Results show that the Gough Map is composed of at least five kinds of dominant red pigments with a particular spatial pattern. This research provides a useful tool for historical geographers and cartographic historians to analyze the material diversity of HSI of cultural heritage artifacts.

Quantification method for the appearance of melanin pigmentation using independent component analysis

NASA Astrophysics Data System (ADS)

Ojima, Nobutoshi; Okiyama, Natsuko; Okaguchi, Saya; Tsumura, Norimichi; Nakaguchi, Toshiya; Hori, Kimihiko; Miyake, Yoichi

2005-04-01

In the cosmetics industry, skin color is very important because skin color gives a direct impression of the face. In particular, many people suffer from melanin pigmentation such as liver spots and freckles. However, it is very difficult to evaluate melanin pigmentation using conventional colorimetric values because these values contain information on various skin chromophores simultaneously. Therefore, it is necessary to extract information of the chromophore of individual skins independently as density information. The isolation of the melanin component image based on independent component analysis (ICA) from a single skin image was reported in 2003. However, this technique has not developed a quantification method for melanin pigmentation. This paper introduces a quantification method based on the ICA of a skin color image to isolate melanin pigmentation. The image acquisition system we used consists of commercially available equipment such as digital cameras and lighting sources with polarized light. The images taken were analyzed using ICA to extract the melanin component images, and Laplacian of Gaussian (LOG) filter was applied to extract the pigmented area. As a result, for skin images including those showing melanin pigmentation and acne, the method worked well. Finally, the total amount of extracted area had a strong correspondence to the subjective rating values for the appearance of pigmentation. Further analysis is needed to recognize the appearance of pigmentation concerning the size of the pigmented area and its spatial gradation.

Chromatographic and spectroscopic identification and recognition of ammoniacal cochineal dyes and pigments

NASA Astrophysics Data System (ADS)

Chieli, A.; Sanyova, J.; Doherty, B.; Brunetti, B. G.; Miliani, C.

2016-06-01

In this work a combined chromatographic and spectroscopic approach is used to provide a diagnostic assessment of semi-synthetic ammoniacal cochineal through the syntheses of its dyes and lakes according to art historical recipes. Commercially introduced in the late XIX century as a dye and pigment, it was used to obtain a brilliant purplish/violet nuance which provided a more stable option over carminic acid although its evidenced use in manufacts and artworks of heritage importance have been scarcely documented. Through HPLC-DAD, it has been possible to identify 4-aminocarminic acid as the main component of ammoniacal cochineal highlighting a chemical formula analogous to acid stable carmine, a recent patented food dye. FTIR clearly distinguishes the amine group in the ammoniacal cochineal dye preparation and TLC-SERS allows for an adequate separation and spectral differentiation in its main components to be evidenced. Colloidal SERS has permitted spectral markers useful in discerning ammoniacal cochineal over carminic acid to be highlighted and discussed. Finally, the methods experimented in this study for the identification of ammoniacal cochineal have been validated on analyzing a sample of dyed wool.

A new mechanistic understanding of light-limitation in the seagrass Zostera muelleri.

PubMed

Davey, Peter A; Pernice, Mathieu; Ashworth, Justin; Kuzhiumparambil, Unnikrishnan; Szabó, Milán; Dolferus, Rudy; Ralph, Peter J

2018-03-01

In this study we investigated the effect of light-limitation (∼20 μmol photons m -2  s -1 ) on the southern hemisphere seagrass, Zostera muelleri. RNA sequencing, chlorophyll fluorometry and HPLC techniques were used to investigate how the leaf-specific transcriptome drives changes in photosynthesis and photo-pigments in Z. muelleri over 6 days. 1593 (7.51%) genes were differentially expressed on day 2 and 1481 (6.98%) genes were differentially expressed on day 6 of the experiment. Differential gene expression correlated with significant decreases in rETR Max , I k , an increase in Yi (initial photosynthetic quantum yield of photosystem II), and significant changes in pigment composition. Regulation of carbohydrate metabolism was observed along with evidence that abscisic acid may serve a role in the low-light response of this seagrass. This study provides a novel understanding of how Z. muelleri responds to light-limitation in the marine water column and provides potential molecular markers for future conservation monitoring efforts. Copyright © 2018 Elsevier Ltd. All rights reserved.

Determination of supplemental feeding needs for astaxanthin and canthaxanthin in salmonids by supramolecular solvent-based microextraction and liquid chromatography-UV/VIS spectroscopy.

PubMed

Caballo, Carmen; Costi, Esther María; Sicilia, María Dolores; Rubio, Soledad

2012-09-15

Development of simple and rapid analytical methods for predicting supplemental feeding requirements in aquaculture is a need to reduce production costs. In this article, a supramolecular solvent (SUPRAS) made up of decanoic acid (DeA) assemblies was proposed to simplify sample treatment in the total and individual determination of carotenoids (red-pink pigments) in farmed salmonids. The analytes were quantitatively extracted in a single step that spends a few minutes using a small volume of SUPRAS (i.e. 800 μL) and directly determined in extracts without the interference from fats or other matrix components. The methods based on the combination of microextraction with SUPRAS and photometry or HPLC-UV/VIS spectroscopy were developed for the determination of total and individual carotenoids, respectively. The applicability of the methods was demonstrated by analysing non-fortified and fortified samples of farmed Atlantic salmons and rainbow trouts. Recoveries obtained by photometry and HPLC-UV/VIS spectroscopy were within the intervals 98-104% and 94-106%, respectively. Copyright © 2012 Elsevier Ltd. All rights reserved.

Application of photoacoustic infrared spectroscopy in the forensic analysis of artists' inorganic pigments.

PubMed

von Aderkas, Eleanor L; Barsan, Mirela M; Gilson, Denis F R; Butler, Ian S

2010-12-01

Fourier-transform photoacoustic infrared (PAIR) spectroscopy has been used in the analysis of 12 inorganic pigments commonly in use by artists today, viz., cobalt blue, ultramarine blue, Prussian blue, azurite, malachite, chromium oxide, viridian, cadmium yellow, chrome yellow, iron oxide, yellow ochre and Mars orange. The authenticity of these 12 commercial pigments was first established by recording their Raman spectra. The subsequent PAIR spectra were highly reproducible and matched well in the mid-IR region with previously published data for these pigments. A number of additional overtone and combination bands were also detected that will prove useful in the identification of the pigments in the future. The PAIR technique is a promising and reliable method for the analysis of inorganic pigments, especially since it involves much simpler preparation than is required for conventional IR measurements. Copyright © 2010. Published by Elsevier B.V.

High-pressure liquid chromatography analysis of antibiotic susceptibility disks.

PubMed Central

Hagel, R B; Waysek, E H; Cort, W M

1979-01-01

The analysis of antibiotic susceptibility disks by high-pressure liquid chromatography (HPLC) was investigated. Methods are presented for the potency determination of mecillinam, ampicillin, carbenicillin, and cephalothin alone and in various combinations. Good agreement between HPLC and microbiological data is observed for potency determinations with recoveries of greater than 95%. Relative standard deviations of lower than 2% are recorded for each HPLC method. HPLC methods offer improved accuracy and greater precision when compared to the standard microbiological methods of analysis for susceptibility disks. PMID:507793

Analysis of Poly-β-Hydroxybutyrate in Rhizobium japonicum Bacteroids by Ion-Exclusion High-Pressure Liquid Chromatography and UV Detection †

PubMed Central

Karr, Dale B.; Waters, James K.; Emerich, David W.

1983-01-01

Ion-exclusion high-pressure liquid chromatography (HPLC) was used to measure poly-β-hydroxybutyrate (PHB) in Rhizobium japonicum bacteroids. The products in the acid digest of PHB-containing material were fractionated by HPLC on Aminex HPX-87H ion-exclusion resin for organic acid analysis. Crotonic acid formed from PHB during acid digestion was detected by its intense absorbance at 210 nm. The Aminex-HPLC method provides a rapid and simple chromatographic technique for routine analysis of organic acids. Results of PHB analysis by Aminex-HPLC were confirmed by gas chromatography and spectrophotometric analysis. PMID:16346443

[18F]MEL050 as a melanin-targeted PET tracer: Fully automated radiosynthesis and comparison to 18F-FDG for the detection of pigmented melanoma in mice primary subcutaneous tumors and pulmonary metastases.

PubMed

Rizzo-Padoin, Nathalie; Chaussard, Michael; Vignal, Nicolas; Kotula, Ewa; Tsoupko-Sitnikov, Vadim; Vaz, Sofia; Hontonnou, Fortune; Liu, Wang-Qing; Poyet, Jean-Luc; Vidal, Michel; Merlet, Pascal; Hosten, Benoit; Sarda-Mantel, Laure

2016-12-01

Melanoma is a highly malignant cutaneous tumor of melanin-producing cells. MEL050 is a synthetic benzamide-derived molecule that specifically binds to melanin with high affinity. Our aim was to implement a fully automated radiosynthesis of [ 18 F]MEL050, using for the first time, the AllInOne™ synthesis module (Trasis), and to evaluate the potential of [ 18 F]MEL050 for the detection of pigmented melanoma in mice primary subcutaneous tumors and pulmonary metastases, and to compare it with that of [ 18 F]FDG. Automated radiosynthesis of [ 18 F]MEL050, including HPLC purification and formulation, were performed on an AllInOne™ synthesis module. [ 18 F]MEL050 was synthesized using a one-step bromine-for-fluorine nucleophilic heteroaromatic substitution. Melanoma models were induced by subcutaneous (primary tumor) or intravenous (pulmonary metastases) injection of B16-F10-luc2 cells in NMRI mice. The maximum percentage of [ 18 F]MEL050 Injected Dose per g of lung tissue (%ID/g Max) was determined on PET images, compared to [ 18 F]FDG and correlated to in vivo bioluminescence imaging. The automated radiosynthesis of [ 18 F]MEL050 required an overall radiosynthesis time of 48min, with a yield of 13-18% (not-decay corrected) and radiochemical purity higher than 99%. [ 18 F]MEL050 PET/CT images were concordant with bioluminescence imaging, showing increased radiotracer uptake in all primary subcutaneous tumors and pulmonary metastases of mice. PET quantification of radiotracers uptake in tumors and muscles demonstrated similar tumor-to-background ratio (TBR) with [ 18 F]MEL050 and [ 18 F]FDG in subcutaneous tumors and higher TBR with [ 18 F]MEL050 than with [ 18 F]FDG in pulmonary metastases. We successfully implemented the radiosynthesis of [ 18 F]MEL050 using the AllInOne™ module, including HPLC purification and formulation. In vivo PET/CT validation of [ 18 F]MEL050 was obtained in mouse models of pigmented melanoma, where higher [ 18 F]MEL050 uptake was observed in sub-millimetric pulmonary metastases, comparatively to [ 18 F]FDG. Copyright © 2016 Elsevier Inc. All rights reserved.

A pigment analysis tool for hyperspectral images of cultural heritage artifacts

NASA Astrophysics Data System (ADS)

Bai, Di; Messinger, David W.; Howell, David

2017-05-01

The Gough Map, in the collection at the Bodleian Library, Oxford University, is one of the earliest surviving maps of Britain. Previous research deemed that it was likely created over the 15th century and afterwards it was extensively revised more than once. In 2015, the Gough Map was imaged using a hyperspectral imaging system at the Bodleian Library. The collection of the hyperspectral image (HSI) data was aimed at faded text enhancement for reading and pigment analysis for the material diversity of its composition and potentially the timeline of its creation. In this research, we introduce several methods to analyze the green pigments in the Gough Map, especially the number and spatial distribution of distinct green pigments. One approach, called the Gram Matrix, has been used to estimate the material diversity in a scene (i.e., endmember selection and dimensionality estimation). Here, we use the Gram Matrix technique to study the within-material differences of pigments in the Gough map with common visual color. We develop a pigment analysis tool that extracts visually common pixels, green pigments in this case, from the Gough Map and estimates its material diversity. It reveals that the Gough Map consists of at least six kinds of dominant green pigments. Both historical geographers and cartographic historians will benefit from this work to analyze the pigment diversity using HSI of cultural heritage artifacts.

Application of principal component analysis to multispectral imaging data for evaluation of pigmented skin lesions

NASA Astrophysics Data System (ADS)

Jakovels, Dainis; Lihacova, Ilze; Kuzmina, Ilona; Spigulis, Janis

2013-11-01

Non-invasive and fast primary diagnostics of pigmented skin lesions is required due to frequent incidence of skin cancer - melanoma. Diagnostic potential of principal component analysis (PCA) for distant skin melanoma recognition is discussed. Processing of the measured clinical multi-spectral images (31 melanomas and 94 nonmalignant pigmented lesions) in the wavelength range of 450-950 nm by means of PCA resulted in 87 % sensitivity and 78 % specificity for separation between malignant melanomas and pigmented nevi.

A new quantitative evaluation method for age-related changes of individual pigmented spots in facial skin.

PubMed

Kikuchi, K; Masuda, Y; Yamashita, T; Sato, K; Katagiri, C; Hirao, T; Mizokami, Y; Yaguchi, H

2016-08-01

Facial skin pigmentation is one of the most prominent visible features of skin aging and often affects perception of health and beauty. To date, facial pigmentation has been evaluated using various image analysis methods developed for the cosmetic and esthetic fields. However, existing methods cannot provide precise information on pigmented spots, such as variations in size, color shade, and distribution pattern. The purpose of this study is the development of image evaluation methods to analyze individual pigmented spots and acquire detailed information on their age-related changes. To characterize the individual pigmented spots within a cheek image, we established a simple object-counting algorithm. First, we captured cheek images using an original imaging system equipped with an illumination unit and a high-resolution digital camera. The acquired images were converted into melanin concentration images using compensation formulae. Next, the melanin images were converted into binary images. The binary images were then subjected to noise reduction. Finally, we calculated parameters such as the melanin concentration, quantity, and size of individual pigmented spots using a connected-components labeling algorithm, which assigns a unique label to each separate group of connected pixels. The cheek image analysis was evaluated on 643 female Japanese subjects. We confirmed that the proposed method was sufficiently sensitive to measure the melanin concentration, and the numbers and sizes of individual pigmented spots through manual evaluation of the cheek images. The image analysis results for the 643 Japanese women indicated clear relationships between age and the changes in the pigmented spots. We developed a new quantitative evaluation method for individual pigmented spots in facial skin. This method facilitates the analysis of the characteristics of various pigmented facial spots and is directly applicable to the fields of dermatology, pharmacology, and esthetic cosmetology. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Atomic force microscopy and scanning electron microscopy analysis of daily disposable limbal ring contact lenses.

PubMed

Lorenz, Kathrine Osborn; Kakkassery, Joseph; Boree, Danielle; Pinto, David

2014-09-01

Limbal ring (also known as 'circle') contact lenses are becoming increasingly popular, especially in Asian markets because of their eye-enhancing effects. The pigment particles that give the eye-enhancing effects of these lenses can be found on the front or back surface of the contact lens or 'enclosed' within the lens matrix. The purpose of this research was to evaluate the pigment location and surface roughness of seven types of 'circle' contact lenses. Scanning electron microscopic (SEM) analysis was performed using a variable pressure Hitachi S3400N instrument to discern the placement of lens pigments. Atomic force microscopy (Dimension Icon AFM from Bruker Nano) was used to determine the surface roughness of the pigmented regions of the contact lenses. Atomic force microscopic analysis was performed in fluid phase under contact mode using a Sharp Nitride Lever probe (SNL-10) with a spring constant of 0.06 N/m. Root mean square (RMS) roughness values were analysed using a generalised linear mixed model with a log-normal distribution. Least square means and their corresponding 95% confidence intervals were estimated for each brand, location and pigment combination. SEM cross-sectional images at 500× and 2,000× magnification showed pigment on the surface of six of the seven lens types tested. The mean depth of pigment for 1-DAY ACUVUE DEFINE (1DAD) lenses was 8.1 μm below the surface of the lens, while the remaining lens types tested had pigment particles on the front or back surface. Results of the atomic force microscopic analysis indicated that 1DAD lenses had significantly lower root mean square roughness values in the pigmented area of the lens than the other lens types tested. SEM and AFM analysis revealed pigment on the surface of the lens for all types tested with the exception of 1DAD. Further research is required to determine if the difference in pigment location influences on-eye performance. © 2014 The Authors. Clinical and Experimental Optometry © 2014 Optometrists Association Australia.

Atomic force microscopy and scanning electron microscopy analysis of daily disposable limbal ring contact lenses

PubMed Central

Lorenz, Kathrine Osborn; Kakkassery, Joseph; Boree, Danielle; Pinto, David

2014-01-01

Background Limbal ring (also known as ‘circle’) contact lenses are becoming increasingly popular, especially in Asian markets because of their eye-enhancing effects. The pigment particles that give the eye-enhancing effects of these lenses can be found on the front or back surface of the contact lens or ‘enclosed’ within the lens matrix. The purpose of this research was to evaluate the pigment location and surface roughness of seven types of ‘circle’ contact lenses. Methods Scanning electron microscopic (SEM) analysis was performed using a variable pressure Hitachi S3400N instrument to discern the placement of lens pigments. Atomic force microscopy (Dimension Icon AFM from Bruker Nano) was used to determine the surface roughness of the pigmented regions of the contact lenses. Atomic force microscopic analysis was performed in fluid phase under contact mode using a Sharp Nitride Lever probe (SNL-10) with a spring constant of 0.06 N/m. Root mean square (RMS) roughness values were analysed using a generalised linear mixed model with a log-normal distribution. Least square means and their corresponding 95% confidence intervals were estimated for each brand, location and pigment combination. Results SEM cross-sectional images at 500× and 2,000× magnification showed pigment on the surface of six of the seven lens types tested. The mean depth of pigment for 1-DAY ACUVUE DEFINE (1DAD) lenses was 8.1 μm below the surface of the lens, while the remaining lens types tested had pigment particles on the front or back surface. Results of the atomic force microscopic analysis indicated that 1DAD lenses had significantly lower root mean square roughness values in the pigmented area of the lens than the other lens types tested. Conclusions SEM and AFM analysis revealed pigment on the surface of the lens for all types tested with the exception of 1DAD. Further research is required to determine if the difference in pigment location influences on-eye performance. PMID:24689948

Demonstration and Evaluation of Solid Phase Microextraction for the Assessment of Bioavailability and Contaminant Mobility

DTIC Science & Technology

2012-05-01

methods demonstrated that desorption into solvents suitable for subsequent chemical analysis (into acetonitrile for HPLC analysis or hexane for GC...SPME. Analysis by HPLC with EPA 8310 with fluorescent detection. a) surface water quality criteria (NRWQC) are given for comparison to detection... analysis ) or hexane (for PCB analysis ) was added to the inserts. The vials were then analyzed directly by HPLC (PAHs) or GC-ECD (PCBs). Fiber achieved

Phytoplankton functional community structure in Argentinian continental shelf determined by HPLC pigment signatures

NASA Astrophysics Data System (ADS)

Moreno, D. Vega; Marrero, J. Pérez; Morales, J.; García, C. Llerandi; Úbeda, M. G. Villagarcía; Rueda, M. J.; Llinás, O.

2012-03-01

The Patagonian Sea in Argentina is known as an area with high primary production and biodiversity. The complex hydrodynamic environment resulting from the interaction between the Malvinas-Brazil convergence and the waters over the continental shelf and slope enhances the development of high chlorophyll concentrations, especially in frontal and coastal areas. The composition, distribution and variability of several phytoplankton functional types (PFTs) derived from diagnostic pigments were studied in relation to the local hydrographical conditions, using data from a research cruise carried out on board the RV Bio Hesperides at the end of the Summer season (March 2008). Phytoplankton cell size and PFT distributions were found to be highly influenced by the physical and chemical characteristics of the studied environments. Thus large cells, mainly diatoms, were the dominant size fraction in the southern frontal areas, where the SubAntarctic Surface Waters (SASW) from the Malvinas Current meet shelf waters. However, other groups of microphytoplankton (mPF), mainly dinoflagellates, were also detected in the shallow waters zone influenced by the tidal regime near the Valdes Peninsula. Picophytoplankton (pPF) was an important contributor to the floristic composition in the southern frontal zones, while nanophytoplankton (nPF) was dominant in the stations located over the continental slope, and in the oligotrophic area near Mar del Plata. The ratio between photoprotective and photosynthetic pigments (PPC:PSC) and the photoprotection index (PI) indirectly provide information about the environment and its effect on the PFTs composition.

Does Uniformity of Topical Corticosteroid Ophthalmic Medications: Flourometholone Acetate 0.1 Suspension and Loteprednol Etabonate 0.5 gel

DTIC Science & Technology

2017-01-03

chromatography ( HPLC ) with photodiode array detection at 240 nm. Results: Flarex® had a mean concentration of 93.7% of the declared concentration when shaken...59 60 Journal of Ocular Pharmacology and Therapeutics Charlton E Stevens Rockville, MD. Methanol, ( HPLC grade), was obtained from Sigma-Aldrich...ST. Louis, MO. HPLC analysis of fluorometholone acetate and loteprednol etabonate HPLC analysis of fluorometholone acetate and loteprednol

Induction of lead-binding phytochelatins in vetiver grass [Vetiveria zizanioides (L.)].

PubMed

Andra, Syam S; Datta, Rupali; Sarkar, Dibyendu; Makris, Konstantinos C; Mullens, Conor P; Sahi, Shivendra V; Bach, Stephan B H

2009-01-01

Elevated lead (Pb) concentrations in residential houseyards around house walls painted with Pb-based pigments pose serious human health risks, especially to children. Vetiver grass (Vetiveria zizanioides L.) has shown promise for use in in situ Pb phytoremediation efforts. However, little is known about the biochemical mechanisms responsible for the observed high Pb tolerance by vetiver. We hypothesized that vetiver exposure to Pb induced the synthesis of phytochelatins (PC(n)) and the formation of Pb-PC(n) complexes, alleviating the phytotoxic effects of free Pb ions. Our main objective was to identify PC(n) and Pb-PC(n) complexes in root and shoot compartments of vetiver grass using high-performance liquid chromatography coupled to electrospray mass spectrometry (HPLC-ES-MS). After 7 d of exposure to Pb, vetiver accumulated up to 3000 mg Pb kg(-1) in shoot tissues, but much higher Pb concentrations were measured in root ( approximately 20,000 mg kg(-1)), without phytotoxic symptoms. Scanning electron micrographs showed Pb deposition in the vascular tissues of root and shoot, suggesting Pb translocation to shoot. Collision-induced dissociation analyses in MS/ MS mode during HPLC-ES-MS analysis allowed for the confirmation of four unique PC(n) (n = 1-4) based on their respective amino acid sequence. The high tolerance of vetiver grass to Pb was attributed to the formation of PC(n) and Pb-PC(n) complexes within the plant tissues, using ES-MS and Pb mass isotopic patterns. These data illustrate the mechanism of high Pb tolerance by vetiver grass, suggesting its potential usefulness for the remediation of Pb-contaminated residential sites.

Micro-Raman analysis of the pigments on painted pottery figurines from two tombs of the Northern Wei Dynasty in Luoyang

NASA Astrophysics Data System (ADS)

Liu, Zhaojun; Han, Yunxia; Han, Ligang; Cheng, Yongjian; Ma, Yiqiang; Fang, Li

2013-05-01

The pigments on the painted pottery figurines from two tombs of Northern Wei Dynasty (AD 386-534) in Luoyang were analyzed by Raman microscopy. All the pigments were identified compared with the Raman spectra of standard pigments. The red pigments were identified as haematite, the blue pigment as lapis lazuli, the green pigment as malachite, the black pigment as carbon black and the white pigment as calcite. Similar pigments were used in the two tombs despite the pottery figurines were very different in artistic style. The use of lapis lazuli as blue pigment on Chinese painted pottery figurines was found for the first time. This pigment and the painted pottery figurine of Sogdians are of great archaeological significance because it demonstrated that the trade and cultural exchanges via the Silk Road had extended to Luoyang city in the Northern Wei Dynasty. The result also confirms that micro-Raman spectroscopy is a powerful analytical method for the identification of pigments on ancient artworks.

Bilirubin present in diverse angiosperms.

PubMed

Pirone, Cary; Johnson, Jodie V; Quirke, J Martin E; Priestap, Horacio A; Lee, David

2010-01-01

Bilirubin is an orange-yellow tetrapyrrole produced from the breakdown of heme by mammals and some other vertebrates. Plants, algae and cyanobacteria synthesize molecules similar to bilirubin, including the protein-bound bilins and phytochromobilin which harvest or sense light. Recently, we discovered bilirubin in the arils of Strelitzia nicolai, the White Bird of Paradise Tree, which was the first example of this molecule in a higher plant. Subsequently, we identified bilirubin in both the arils and the flowers of Strelitzia reginae, the Bird of Paradise Flower. In the arils of both species, bilirubin is present as the primary pigment, and thus functions to produce colour. Previously, no tetrapyrroles were known to generate display colour in plants. We were therefore interested in determining whether bilirubin is broadly distributed in the plant kingdom and whether it contributes to colour in other species. In this paper, we use HPLC/UV and HPLC/UV/electrospray ionization-tandem mass spectrometry (HPLC/UV/ESI-MS/MS) to search for bilirubin in 10 species across diverse angiosperm lineages. Bilirubin was present in eight species from the orders Zingiberales, Arecales and Myrtales, but only contributed to colour in species within the Strelitziaceae. The wide distribution of bilirubin in angiosperms indicates the need to re-assess some metabolic details of an important and universal biosynthetic pathway in plants, and further explore its evolutionary history and function. Although colour production was limited to the Strelitziaceae in this study, further sampling may indicate otherwise.

Bilirubin present in diverse angiosperms

PubMed Central

Pirone, Cary; Johnson, Jodie V.; Quirke, J. Martin E.; Priestap, Horacio A.; Lee, David

2010-01-01

Background and aims Bilirubin is an orange-yellow tetrapyrrole produced from the breakdown of heme by mammals and some other vertebrates. Plants, algae and cyanobacteria synthesize molecules similar to bilirubin, including the protein-bound bilins and phytochromobilin which harvest or sense light. Recently, we discovered bilirubin in the arils of Strelitzia nicolai, the White Bird of Paradise Tree, which was the first example of this molecule in a higher plant. Subsequently, we identified bilirubin in both the arils and the flowers of Strelitzia reginae, the Bird of Paradise Flower. In the arils of both species, bilirubin is present as the primary pigment, and thus functions to produce colour. Previously, no tetrapyrroles were known to generate display colour in plants. We were therefore interested in determining whether bilirubin is broadly distributed in the plant kingdom and whether it contributes to colour in other species. Methodology In this paper, we use HPLC/UV and HPLC/UV/electrospray ionization-tandem mass spectrometry (HPLC/UV/ESI-MS/MS) to search for bilirubin in 10 species across diverse angiosperm lineages. Principal results Bilirubin was present in eight species from the orders Zingiberales, Arecales and Myrtales, but only contributed to colour in species within the Strelitziaceae. Conclusions The wide distribution of bilirubin in angiosperms indicates the need to re-assess some metabolic details of an important and universal biosynthetic pathway in plants, and further explore its evolutionary history and function. Although colour production was limited to the Strelitziaceae in this study, further sampling may indicate otherwise. PMID:22476078

Quality Analysis of Chlorogenic Acid and Hyperoside in Crataegi fructus

PubMed Central

Weon, Jin Bae; Jung, Youn Sik; Ma, Choong Je

2016-01-01

Background: Crataegi fructus is a herbal medicine for strong stomach, sterilization, and alcohol detoxification. Chlorogenic acid and hyperoside are the major compounds in Crataegi fructus. Objective: In this study, we established novel high-performance liquid chromatography (HPLC)-diode array detection analysis method of chlorogenic acid and hyperoside for quality control of Crataegi fructus. Materials and Methods: HPLC analysis was achieved on a reverse-phase C18 column (5 μm, 4.6 mm × 250 mm) using water and acetonitrile as mobile phase with gradient system. The method was validated for linearity, precision, and accuracy. About 31 batches of Crataegi fructus samples collected from Korea and China were analyzed by using HPLC fingerprint of developed HPLC method. Then, the contents of chlorogenic acid and hyperoside were compared for quality evaluation of Crataegi fructus. Results: The results have shown that the average contents (w/w %) of chlorogenic acid and hyperoside in Crataegi fructus collected from Korea were 0.0438% and 0.0416%, respectively, and the average contents (w/w %) of 0.0399% and 0.0325%, respectively. Conclusion: In conclusion, established HPLC analysis method was stable and could provide efficient quality evaluation for monitoring of commercial Crataegi fructus. SUMMARY Quantitative analysis method of chlorogenic acid and hyperoside in Crataegi fructus is developed by high.performance liquid chromatography.(HPLC).diode array detectionEstablished HPLC analysis method is validated with linearity, precision, and accuracyThe developed method was successfully applied for quantitative analysis of Crataegi fructus sample collected from Korea and China. Abbreviations used: HPLC: High-performance liquid chromatography, GC: Gas chromatography, MS: Mass spectrometer, LOD: Limits of detection, LOQ: Limits of quantification, RSD: Relative standard deviation, RRT: Relative retention time, RPA: Relation peak area. PMID:27076744

The Comparison Between Nmf and Ica in Pigment Mixture Identification of Ancient Chinese Paintings

NASA Astrophysics Data System (ADS)

Liu, Y.; Lyu, S.; Hou, M.; Yin, Q.

2018-04-01

Since the colour in painting cultural relics observed by our naked eyes or hyperspectral cameras is usually a mixture of several kinds of pigments, the mixed pigments analysis will be an important subject in the field of ancient painting conservation and restoration. This paper aims to find a more effective method to confirm the types of every pure pigment from mixture on the surface of paintings. Firstly, we adopted two kinds of blind source separation algorithms, which are independent component analysis and non-negative matrix factorization, to extract the pure pigment component from mixed spectrum respectively. Moreover, we matched the separated pure spectrum with the pigments spectra library built by our team to determine the pigment type. Furthermore, three kinds of data including simulation data, mixed pigments spectral data measured in laboratory, and the spectral data of an ancient painting were chosen to evaluate the performance of the different algorithms. And the accuracy was compared between the two algorithms. Finally, the experimental results show that non-negative matrix factorization method is more suitable for endmember extraction in the field of ancient painting conservation and restoration.

Daily changes of structure, function and rhodopsin content in the compound eye of the crab Hemigrapsus sanguineus.

PubMed

Arikawa, K; Kawamata, K; Suzuki, T; Eguchi, E

1987-08-01

The compound eye of the crab hemigrapsus sanguineus undergoes daily changes in morphology as determined by light and electron microscopy, both in the quantity of chromophore substances studied by HPLC and in visual sensitivity as shown by electrophysiological techniques. 1. At a temperature of 20 degrees C, the rhabdom occupation ratio (ROR) of an ommatidial retinula was 11.6% (maximum) at midnight, 8.0 times larger than the minimum value at midday (1.4%). 2. Observations by freeze-fracture revealed that the densities of intra-membranous particles (9-11 nm in diameter) of rhabdomeric membrane were ca. 2000/microns 2 and ca. 3000/microns 2 for night and daytime compound eyes, respectively. 3. Screening pigment granules migrated longitudinally and aggregated at night, but dispersed during the day. Reflecting pigment granules migrate transversally in the proximal half of the reticula layer i.e. cytoplasmic extensions containing reflecting pigment granules squeeze between neighbouring retinula cells causing optical isolation (Fig. 4). Thus the screening pigment granules within the retinula cells show longitudinal migration and radial movement so that the daytime rhabdoms are closely surrounded by the pigment granules. 4. At 20 degrees C, the total amount of chromophore of the visual pigment (11-cis and all-trans-retinal) was 1.4 times larger at night than during the day i.e. 46.6 pmol/eye at midnight and 33.2 pmol/eye at midday. Calculations of the total surface area of rhabdomeric membrane, total number of intra-membranous particles in rhabdomeric membrane and the total number of chromophore molecules in a compound eye, indicate that a considerable amount of chromophore-protein complex exists outside the rhabdom during the day. 5. The change in rhabdom size and quantity of chromophore were highly dependent on temperature. At 10 degrees C both rhabdom size and amount of chromophore stayed close to daytime levels throughout the 24 hours. 6. The intracellularly determined relative sensitivity of the dark adapted night eye to a point source of light was about twice as high as the dark-adapted day eye. Most of the increase in the sensitivity is attributed primarily to the effect of reflecting pigment migration around the basement membrane and, secondarily, to the changes in the amount and properties of the photoreceptive membrane. The results form the basis of a detailed discussion as to how an apposition eye can function possibly as a night-eye.

Lipophilization and MS characterization of the main anthocyanins purified from hibiscus flowers.

PubMed

Grajeda-Iglesias, Claudia; Salas, Erika; Barouh, Nathalie; Baréa, Bruno; Figueroa-Espinoza, Maria Cruz

2017-09-01

Hibiscus sabdariffa flowers represent an interesting source of anthocyanins, one of the most important plant pigments, which are responsible of the intense red color of the calyces, and have potential as natural colorants for food applications. Nevertheless, anthocyanins are highly hydrosoluble and unstable compounds. On this basis, the aim of this work was to increase the lipophilicity of the hibiscus anthocyanins by lipophilization, in order to obtain amphiphilic colorants, which could be easily incorporated in lipid-rich food matrices. Octanoyl derivatives of delphinidin-3-O-sambubioside and cyanidin-3-O-sambubioside were chemically obtained for the first time, and characterized by means of HPLC-ESI-MS data. Copyright © 2017. Published by Elsevier Ltd.

Determination of Nitroaromatic, Nitramine, and Nitrate Ester Explosives in Soils Using GC-ECD

DTIC Science & Technology

1999-08-01

for supplying soils from minefields; and Dr. Paul H. Miyares, CRREL, for HPLC analysis of Fort Leonard Wood soil extracts. ii CONTENTS P reface...42 ILLUSTRATIONS Figure 1. Correlation analysis of GC-ECD concentration (mg/kg) estimates with those from HPLC -UV...kg) estimates with those from HPLC -UV analysis using splits of the same acetonitrile extract from archived soils

The use of capillary electrophoresis as part of a specificity testing strategy for mitoguazone dihydrochloride HPLC methods.

PubMed

Thomson, C E; Gray, M R; Baxter, M P

1997-05-01

Capillary electrophoresis (CE) has been used as part of a validation experiment designed to prove the specificity of high performance liquid chromatography (HPLC) methods used for analysis of mitoguazone dihydrochloride drug substance. Data regarding accuracy, precision and sensitivity of the CE methods are presented as well as a comparison of results obtained from CE, HPLC and thin-layer chromatography (TLC) analysis of samples stressed under a variety of conditions. It was concluded that, not only were the HPLC methods being investigated specific, but that CE could potentially be used to replace HPLC for the routine assay of mitoguazone dihydrochloride.

Characterizing pigments with hyperspectral imaging variable false-color composites

NASA Astrophysics Data System (ADS)

Hayem-Ghez, Anita; Ravaud, Elisabeth; Boust, Clotilde; Bastian, Gilles; Menu, Michel; Brodie-Linder, Nancy

2015-11-01

Hyperspectral imaging has been used for pigment characterization on paintings for the last 10 years. It is a noninvasive technique, which mixes the power of spectrophotometry and that of imaging technologies. We have access to a visible and near-infrared hyperspectral camera, ranging from 400 to 1000 nm in 80-160 spectral bands. In order to treat the large amount of data that this imaging technique generates, one can use statistical tools such as principal component analysis (PCA). To conduct the characterization of pigments, researchers mostly use PCA, convex geometry algorithms and the comparison of resulting clusters to database spectra with a specific tolerance (like the Spectral Angle Mapper tool on the dedicated software ENVI). Our approach originates from false-color photography and aims at providing a simple tool to identify pigments thanks to imaging spectroscopy. It can be considered as a quick first analysis to see the principal pigments of a painting, before using a more complete multivariate statistical tool. We study pigment spectra, for each kind of hue (blue, green, red and yellow) to identify the wavelength maximizing spectral differences. The case of red pigments is most interesting because our methodology can discriminate the red pigments very well—even red lakes, which are always difficult to identify. As for the yellow and blue categories, it represents a good progress of IRFC photography for pigment discrimination. We apply our methodology to study the pigments on a painting by Eustache Le Sueur, a French painter of the seventeenth century. We compare the results to other noninvasive analysis like X-ray fluorescence and optical microscopy. Finally, we draw conclusions about the advantages and limits of the variable false-color image method using hyperspectral imaging.

Demonstration and Evaluation of Solid Phase Microextraction for the Assessment of Bioavailability and Contaminant Mobility. ESTCP Cost and Performance Report

DTIC Science & Technology

2012-08-01

subsequent chemical analysis (into acetonitrile for high-performance liquid chromatography [ HPLC ] analysis or hexane for gas chromatography [GC... analysis ) is rapid and complete. In this work, PAHs were analyzed by Waters 2795 HPLC with fluorescent detection (USEPA Method 8310) and PCBs were...detection limits by direct water injection versus SPME with PDMS and coefficient of variation and correlation coefficient for SPME. Analysis by HPLC

Differentiation of Pigment in Eggs Using Carbon ((13)C/(12)C) and Nitrogen ((15)N/(14)N) Stable Isotopes.

PubMed

Sun, Feng M; Shi, Guang Y; Wang, Hui W

2016-07-01

Consumers prefer natural and healthy food, but artificial pigments are often abused in egg products. The study aimed at differentiating the origin of pigments in eggs by applying the technique of carbon ((13)C/(12)C) and nitrogen ((15)N/(14)N) stable isotope analysis. Five hundred sixty laying hens were randomly distributed into 14 treatments, which were divided into four groups: maize, carophyll red pigment, carophyll yellow pigment, and a mixture of carophyll red and yellow pigments. Eggs were collected and pretreated to determe the values of the Roche Yolk Color Fan (RCF), δ(13)C, and δ(15)N. With increasing maize content, the RCF and δ(13)C values of yolks increased. Moreover, the RCF values in the three pigment groups were significantly influenced by the artificial colors, but δ(13)C values were not significantly different, regardless of the existence of pigment. The δ(15)N values in all treatments did not vary as regularly as the carbon stable isotope. A strong positive correlation was found between RCF and δ(13)C in the maize group, but no such correlation was be observed in the pigment groups. It is concluded that carbon stable isotope ratio analysis (δ(13)C) of the yolk can be used to differentiate the origin of the pigment added to eggs.

Verification of rapid method for estimation of added food colorant type in boiled sausages based on measurement of cross section color

NASA Astrophysics Data System (ADS)

Jovanović, J.; Petronijević, R. B.; Lukić, M.; Karan, D.; Parunović, N.; Branković-Lazić, I.

2017-09-01

During the previous development of a chemometric method for estimating the amount of added colorant in meat products, it was noticed that the natural colorant most commonly added to boiled sausages, E 120, has different CIE-LAB behavior compared to artificial colors that are used for the same purpose. This has opened the possibility of transforming the developed method into a method for identifying the addition of natural or synthetic colorants in boiled sausages based on the measurement of the color of the cross-section. After recalibration of the CIE-LAB method using linear discriminant analysis, verification was performed on 76 boiled sausages, of either frankfurters or Parisian sausage types. The accuracy and reliability of the classification was confirmed by comparison with the standard HPLC method. Results showed that the LDA + CIE-LAB method can be applied with high accuracy, 93.42 %, to estimate food color type in boiled sausages. Natural orange colors can give false positive results. Pigments from spice mixtures had no significant effect on CIE-LAB results.

High throughput screening and profiling of high-value carotenoids from a wide diversity of bacteria in surface seawater.

PubMed

Asker, Dalal

2018-09-30

Carotenoids are valuable natural colorants that exhibit numerous health promoting properties, and thus are widely used in food, feeds, pharmaceutical and nutraceuticals industries. In this study, we isolated and identified novel microbial sources that produced high-value carotenoids using high throughput screening (HTS). A total of 701 pigmented microbial strains library including marine bacteria and red yeast was constructed. Carotenoids profiling using HPLC-DAD-MS methods showed 88 marine bacterial strains with potential for the production of high-value carotenoids including astaxanthin (28 strains), zeaxanthin (21 strains), lutein (1 strains) and canthaxanthin (2 strains). A comprehensive 16S rRNA gene based phylogenetic analysis revealed that these strains can be classified into 30 species belonging to five bacterial classes (Flavobacteriia, α-Proteobacteria, γ-Proteobacteria, Actinobacteria and Bacilli). Importantly, we discovered novel producers of zeaxanthin and lutein, and a high diversity in both carotenoids and producing microbial strains, which are promising and highly selective biotechnological sources for high-value carotenoids. Copyright © 2018 Elsevier Ltd. All rights reserved.

A Novel Epiphytic Chlorophyll d-containing Cyanobacterium Isolated from a Mangrove-associated Red Alga.

PubMed

Larkum, Anthony W D; Chen, Min; Li, Yaqiong; Schliep, Martin; Trampe, Erik; West, John; Salih, Anya; Kühl, Michael

2012-12-01

A new habitat and a new chlorophyll (Chl) d-containing cyanobacterium belonging to the genus Acaryochloris are reported in this study. Hyperspectral microscopy showed the presence of Chl d-containing microorganisms in epiphytic biofilms on a red alga (Gelidium caulacantheum) colonizing the pneumato-phores of a temperate mangrove (Avicennia marina). The presence of Chl d was further proven by high performance liquid chromatography (HPLC)-based pigment analysis and by confocal imaging of cultured cells. Enrichment of mangrove biofilm samples under near-infrared radiation (NIR) yielded the new Acaryochloris sp. MPGRS1, which was closely related in terms of 16S rRNA gene sequence to an isolate from the hypertrophic Salton Sea, USA. The new isolate used Chl d as its major photopigment; Chl d and Chl a contents were ~98% and 1%-2% of total cellular chlorophyll, respectively. These findings expand the variety of ecological niches known to harbor Chl d-containing cyanobacteria and support our working hypothesis that such oxyphototrophs may be ubiquitous in habitats depleted of visible light, but with sufficient NIR exposure. © 2012 Phycological Society of America.

Qualitative and Quantitative Analysis of Flower Pigments in Chocolate Cosmos, Cosmos atrosanguineus, and its Hybrids.

PubMed

Amamiya, Kotarou; Iwashina, Tsukasa

2016-01-01

Two major anthocyanins, cyanidin 3-O-glucoside and 3-O-rutinoside, were isolated from the black flowers of Cosmos atrosanguineus cultivar 'Choco Mocha', together with three minor anthocyanins, cyanidin 3-O-malonylglucoside, pelargonidin 3-O-glucoside and 3-O-rutinoside. A chalcone, butein 4'-O-glucoside and three minor flavanones were isolated from the red flowers of C. atrosanguineis x C. sulphureus cultivar 'Rouge Rouge'. The anthocyanins and chalcone accumulation of cultivar 'Choco Mocha' and its hybrid cultivars 'Brown Rouge', 'Forte Rouge', 'Rouge Rouge' and 'Noel Rouge' was surveyed by quantitative HPLC. Total anthocyanins of black flower cultivars 'Choco Mocha' and 'Brown Rouge' were 3-4-folds higher than that of the red flower cultivar 'Noel Rouge'. On the other hand, total chalcone of 'Noel Rouge' was 10-77-folds higher compared with those of other cultivars, 'Brown Rouge', 'Forte Rouge' and 'Rouge Rouge'. It was shown that the flower color variations from red to black of Chocolate Cosmos and its hybrids are due to the difference in the relative amounts of anthocyanins and chalcone.

Qualitative and quantitative analysis of the diuretic component ergone in Polyporus umbellatus by HPLC with fluorescence detection and HPLC-APCI-MS/MS.

PubMed

Zhao, Ying-Yong; Zhao, Ye; Zhang, Yong-Min; Lin, Rui-Chao; Sun, Wen-Ji

2009-06-01

Polyporus umbellatus is a widely used anti-aldosteronic diuretic in Traditional Chinese medicine (TCM). A new, sensitive and selective high-performance liquid chromatography-fluorescence detector (HPLC-FLD) and high-performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (HPLC-APCI-MS/MS) method for quantitative and qualitative determination of ergosta-4,6,8(14),22-tetraen-3-one(ergone), which is the main diuretic component, was provided for quality control of P. umbellatus crude drug. The ergone in the ethanolic extract of P. umbellatus was unambiguously characterized by HPLC-APCI, and further confirmed by comparing with a standard compound. The trace ergone was detected by the sensitive and selective HPLC-FLD. Linearity (r2 > 0.9998) and recoveries of low, medium and high concentration (100.5%, 100.2% and 100.4%) were consistent with the experimental criteria. The limit of detection (LOD) of ergone was around 0.2 microg/mL. Our results indicated that the content of ergone in P. umbellatus varied significantly from habitat to habitat with contents ranging from 2.13 +/- 0.02 to 59.17 +/- 0.05 microg/g. Comparison among HPLC-FLD and HPLC-UV or HPLC-APCI-MS/MS demonstrated that the HPLC-FLD and HPLC-APCI-MS/MS methods gave similar quantitative results for the selected herb samples, the HPLC-UV methods gave lower quantitative results than HPLC-FLD and HPLC-APCI-MS/MS methods. The established new HPLC-FLD method has the advantages of being rapid, simple, selective and sensitive, and could be used for the routine analysis of P. umbellatus crude drug.

Sulfate as a pivotal factor in regulation of Serratia sp. strain S2B pigment biosynthesis.

PubMed

Rastegari, Banafsheh; Karbalaei-Heidari, Hamid Reza

2016-10-01

In the present work, we investigated the prodiginine family as secondary metabolite members. Bacterial strain S2B, with the ability to produce red pigment, was isolated from the Sarcheshmeh copper mine in Iran. 16S rDNA gene sequencing revealed that the strain was placed in the Serratia genus. Pigment production was optimized using low-cost culture medium and the effects of various physicochemical factors were studied via statistical approaches. Purification of the produced pigment by silica gel column chromatography showed a strong red pigment fraction and a weaker orange band. Mass spectrometry, FT-IR spectroscopy and (1)H NMR analysis revealed that the red pigment was prodigiosin and the orange band was a prodigiosin-like analog, with molecular weights of 323 and 317 Da, respectively. Genotoxicity and cytotoxicity studies confirmed their membership in the prodiginine family. Analysis of the production pattern of the pigments in the presence of different concentrations of ammonium salts revealed the role of sulfate as an important factor in regulation of the pigment biosynthesis pathway. Overall, the data showed that regulation of the pigment biosynthesis pathway in Serratia sp. strain S2B was affected by inorganic micronutrients, particularly the sulfate ions. Copyright © 2016 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Chromatographic and spectroscopic identification and recognition of ammoniacal cochineal dyes and pigments.

PubMed

Chieli, A; Sanyova, J; Doherty, B; Brunetti, B G; Miliani, C

2016-06-05

In this work a combined chromatographic and spectroscopic approach is used to provide a diagnostic assessment of semi-synthetic ammoniacal cochineal through the syntheses of its dyes and lakes according to art historical recipes. Commercially introduced in the late XIX century as a dye and pigment, it was used to obtain a brilliant purplish/violet nuance which provided a more stable option over carminic acid although its evidenced use in manufacts and artworks of heritage importance have been scarcely documented. Through HPLC-DAD, it has been possible to identify 4-aminocarminic acid as the main component of ammoniacal cochineal highlighting a chemical formula analogous to acid stable carmine, a recent patented food dye. FTIR clearly distinguishes the amine group in the ammoniacal cochineal dye preparation and TLC-SERS allows for an adequate separation and spectral differentiation in its main components to be evidenced. Colloidal SERS has permitted spectral markers useful in discerning ammoniacal cochineal over carminic acid to be highlighted and discussed. Finally, the methods experimented in this study for the identification of ammoniacal cochineal have been validated on analyzing a sample of dyed wool. Copyright © 2016 Elsevier B.V. All rights reserved.

Porphyrins produce uniquely ephemeral animal colouration: a possible signal of virginity

NASA Astrophysics Data System (ADS)

Galván, Ismael; Camarero, Pablo R.; Mateo, Rafael; Negro, Juan J.

2016-12-01

Colours that underlie animal pigmentation can either be permanent or renewable in the short term. Here we describe the discovery of a conspicuous salmon-pink colouration in the base of bustard feathers and down that has never been reported because of its extraordinarily brief expression. HPLC analyses indicated that its constituent pigments are coproporphyrin III and protoporphyrin IX, which are prone to photodegradation. Accordingly, an experimental exposure of feathers of three bustard species to sunlight produced a rapid disappearance of the salmon-pink colouration, together with a marked decrease in reflectance around 670 nm coinciding with the absorption of porphyrin photoproducts. The disappearance of the salmon-pink colouration can occur in a period as short as 12 min, likely making it the most ephemeral colour phenotype in any extant bird. The presence of this colour trait in males performing sexual displays may thus indicate to females a high probability that the males were performing their first displays and would engage in their first copulations in the breeding season. In dominant males, sperm quality decreases over successive copulations, thus porphyrin-based colouration may evolve as a signal of virginity that allows females to maximize their fitness in lek mating systems.

[Carotenoids and vitamin A in fish].

PubMed

Elmadfa, I; Majchrzak, D

1998-06-01

Seven commercial salmon and six trout samples were investigated. Retinol and the carotenoids astaxanthin and canthaxanthin important for pigmentation of the muscle were determined by RP-HPLC. Vitamin A concentrations of raw salmon samples were 16-19 micrograms/100 g, of smoked salmon samples 9-19 micrograms/ 100 g; retinol values of salmon trout (raw) and trout (raw) reached 14-16 micrograms/100 g and 7-9 micrograms/100 g. Concentrations of astaxanthin the important carotenoid of pigmentation, ranged in samples of salmon from 310-465 micrograms/100 g. Samples of salmon trout showed astaxanthin values between 90 and 536 micrograms/100 g, trout samples only 67-85 micrograms/100 g. Concentrations of canthaxanthin were different in the examined samples and were not detectable in all samples. Highest values of canthaxanthin were found in salmon trout samples (113-226 micrograms/100 g), Irish smoked salmon and stremel-salmon (145-169 micrograms/100 g). Raw samples of salmon and of trout showed only low concentrations of canthaxanthin. Astaxanthin and canthaxanthin together reached values of 419-524 micrograms/100 g for salmon, 316-701 micrograms/100 g for salmon trout, and 72-91 micrograms/100 g for trout samples.

Identification and quantification of Cu-chlorophyll adulteration of edible oils.

PubMed

Fang, Mingchih; Tsai, Chia-Fen; Wu, Guan-Yan; Tseng, Su-Hsiang; Cheng, Hwei-Fang; Kuo, Ching-Hao; Hsu, Che-Lun; Kao, Ya-Min; Shih, Daniel Yang-Chih; Chiang, Yu-Mei

2015-01-01

Cu-pyropheophytin a, the major Cu-pigment of Cu-chlorophyll, was determined in edible oil by high-resolution mass spectrometry with a high-performance liquid chromatography-quadrupole (HPLC-Q)-Orbitrap system and by HPLC coupled with a photodiode-array detector. Respective limit of detection and limit of quantification levels of 0.02 μg/g and 0.05 μg/g were obtained. Twenty-nine commercial oil products marked as olive oil, grapeseed oil and blended oil, all sourced directly from a food company that committed adulteration with Cu-chlorophyll, were investigated. In this company, four green dyes illegally used in oils were seized during factory investigation by the health authorities. The food additive Cu-pyropheophytin a was found in all confiscated samples in concentrations between 0.02 and 0.39 μg/g. Survey results of another 235 commercial oil samples manufactured from other companies, including olive pomace oil, extra virgin olive oil, olive oil, grapeseed oil and blended oil, indicated high positive incidences of 63%, 39%, 44%, 97% and 8%, respectively, with a concentration range between 0.02 and 0.54 μg/g. High Cu-chlorophyll concentrations are indications for fraudulent adulteration of oils.

Optical sensing of concentration and refractive index of pigments in a suspension.

PubMed

Niskanen, Ilpo; Räty, Jukka; Peiponen, Kai-Erik

2010-06-10

We describe an immersion liquid and data analysis method for the simultaneous determination of the refractive index and concentration of pigments by measurement of light transmission of suspensions. A new innovation is that, in the event that two different pigments are simultaneously present in a suspension, it is possible to detect the refractive index of an unknown pigment with the aid of half-width of transmittance and, furthermore, to obtain the concentration of the unknown pigment.

Carotenoids in the Gulf of Gdansk sediments- useful markers of environmental conditions in the past

NASA Astrophysics Data System (ADS)

Krajewska, Magdalena; Szymczak-Żyła, Małgorzata; Kowalewska, Grażyna

2017-04-01

Carotenoids are a large group of natural compounds widespread in the aquatic environment. Most of carotenoids in sediments originate from phytoplankton, macroalgae, vascular plants and bacteria. Carotenoids undergo different reactions in water column and after deposition in sediments. Concentration and relative composition of pigments in sediments depend on such factors like primary production, phytoplankton taxonomy, sedimentation and accumulation rate, hydrological and post-depositional conditions. Because some pigments are unstable and can be degraded both by abiotic and biotic factors - in the presence of light, oxygen, herbivores or microorganisms activity, they provide information about conditions in water column and in sediments. They differ in stability and, due to that, carotenoids in marine sediments are indicators, not only of organic matter sources but also of pre- and post-depositional conditions. This work presents a concentration and distribution of selected carotenoids in recent (6 cores 0-20 cm) and deep (1 core, up to 400 cm) sediments of the Gulf of Gdansk- a highly eutrophic area of high primary production and high sedimentation rate. The sediments were collected during two cruises and analysed in framework of CLISED ('Climate Change Impact on Ecosystem Health- Marine Sediment Indicators') Polish- Norwegian research Project, grant no. 196128. Just after collection, the samples were frozen and kept in such a state until analysis in land laboratory. There, after extraction, carotenoids were analysed using high performance liquid chromatography (HPLC-DAD). Sediment age has been defined using C-14 dating. Sediments contained parent carotenoids, markers of the main phytoplankton groups occurring in the Baltic, e.g. diatoms, green algae and cyanobacteria. B-carotene in sediments is a better, averaged, marker of primary production than chlorophyll- a and similarly stable one as sum of chloropigments-a. Presentation will focus on cyanobacteria and their pigments record in sediments, because cyanobacterial blooms are good indicators of eutrophication and anaerobic conditions in the past. Carotenoids record in deep sediments was varied and there are periods of better or worse carotenoids preservation which proves climate changes.

Photosynthesis, chlorophyll fluorescence, light-harvesting system and photoinhibition resistance of a zeaxanthin-accumulating mutant of Arabidopsis thaliana.

PubMed

Tardy, F; Havaux, M

1996-06-01

The abscisic-acid-deficient aba-1 mutant of Arabidopsis thaliana is unable to epoxidize zeaxanthin. As a consequence, it contains large amounts of this carotenoid and lacks epoxy-xanthophylls. HPLC analysis of pigment contents in leaves, isolated thylakoids and preparations of the major light-harvesting complex of photosystem II (PSII) (LHC-II) indicated that zeaxanthin replaced neoxanthin, violaxanthin and antheraxanthin in the light-harvesting system of PSII in aba-1. Non-denaturing electrophoretic fractionation of solubilized thylakoids showed that the xanthophyll imbalance in aba-1 was associated with a pronounced decrease in trimeric LHC-II in favour of monomeric complexes, with a substantial increase in free pigments (mainly zeaxanthin and chlorophyll b), suggesting a decreased stability of LHC-II. The reduced thermostability of PSII in aba-1 was also deduced from in vivo chlorophyll fluorescence measurements. Wild-type and aba-1 leaves could not be distinguished on the basis of their photosynthetic performance: no significant difference was observed between the two types of leaves for light-limited and light-saturated photosynthetic oxygen evolution, PSII photochemistry and PSII to PSI electron flow. When dark-adapted leaves (grown in white light of 80 mumol m-2s-1) were suddenly exposed to red light of 150 mumol m-2s-1, there was a strong nonphotochemical quenching of chlorophyll fluorescence, the amplitude of which was virtually identical (at steady state) in aba-1 and wild-type leaves, despite the fact that the xanthophyll cycle pigment pool was completely in the form of zeaxanthin in aba-1 and almost exclusively in the form of violaxanthin in the wild type. A high concentration of zeaxanthin in aba-1 thylakoids did not, in itself, provide any particular protection against the photoinhibition of PSII. Taken together, the presented results indicate the following: (1) zeaxanthin can replace epoxy-xanthophylls in LHC-II without significantly affecting the photochemical efficiency of PSII; (2) zeaxanthin does not play any specific role in direct (thermal) energy dissipation in PSII; (3) the photoprotective action of the xanthophyll cycle (rapid photoconversion of violaxanthin to zeaxanthin) is not based on the mere substitution of violaxanthin by zeaxanthin in the chlorophyll antennae.

Comparative HPLC/ESI-MS and HPLC/DAD study of different populations of cultivated, wild and commercial Gentiana lutea L.

PubMed

Mustafa, Ahmed M; Caprioli, Giovanni; Ricciutelli, Massimo; Maggi, Filippo; Marín, Rosa; Vittori, Sauro; Sagratini, Gianni

2015-05-01

The root of Gentiana lutea L., famous for its bitter properties, is often used in alcoholic bitter beverages, food products and traditional medicine to stimulate the appetite and improve digestion. This study presents a new, fast, and accurate HPLC method using HPLC/ESI-MS and HPLC/DAD for simultaneous analysis of iridoids (loganic acid), secoiridoids (gentiopicroside, sweroside, swertiamarin, amarogentin) and xanthones (isogentisin) in different populations of G.lutea L., cultivated in the Monti Sibillini National Park, obtained wild there, or purchased commercially. Comparison of HPLC/ESI-MS and HPLC/DAD indicated that HPLC/ESI-MS is more sensitive, reliable and selective. Analysis of twenty samples showed that gentiopicroside is the most dominant compound (1.85-3.97%), followed by loganic acid (0.11-1.30%), isogentisin (0.03-0.48%), sweroside (0.05-0.35%), swertiamarin (0.08-0.30%), and amarogentin (0.01-0.07%). The results confirmed the high quality of the G.lutea cultivated in the Monti Sibillini National Park. Copyright © 2014 Elsevier Ltd. All rights reserved.

Forensic applications of direct analysis in real time (DART) coupled to Q-orbitrap tandem mass spectrometry for the in situ analysis of pigments from paint evidence.

PubMed

Chen, Tai-Hung; Wu, Shu-Pao

2017-08-01

The accurate examination of paint fragments obtained from an accident, such as those obtained from vehicles involved in a hit-and-run case, is often critical in forensic investigations. However, organic pigments are typically minor components of automotive coatings, which makes discrimination difficult. In this study, direct analysis in real time coupled to Q-orbitrap tandem mass spectrometry (DART-MS) was employed to detect a wide range of common organic pigments in vehicle paints. Twelve common organic pigments used in vehicle paints, such as red, yellow, orange, and purple, were tested, and a database was constructed for future examinations of vehicle paint. Two hit-and-run vehicle accident cases, which occurred in New Taipei City, were investigated by Fourier transform infrared (FTIR) spectroscopy and DART-MS. First, FTIR spectroscopy was employed to study the paint samples as a preliminary screening step. Most of the observed IR peaks were attributed to binder and extenders present in paints. The IR peaks corresponding to the organic pigments were found to be weak and overlapped with those corresponding to resins. On the other hand, DART-MS successfully characterized the organic pigments. DART-MS was found to be excellent for rapidly determining the presence of organic pigments in paint samples without the need for a complicated pre-treatment process or lengthy analysis time. Copyright © 2017 Elsevier B.V. All rights reserved.

Analytical Investigation Of Pigments, Ground Layer And Media Of Cartonnage Fragments From Greek Roman Period

NASA Astrophysics Data System (ADS)

Afifi, Hala. A. M.

Some cartonnage fragments from Hawara, Fayoum Excavation were examined to identify pigments, media and grounds. It belonged to the Greek-Roman period. They were studied by X-ray diffraction (XRD), Energy dispersive X ray analysis (EDS) equipped with Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). These techniques were used to identify the composition and morphology of grounds, nature of pigments and media used in cartonnage fragments. The coarse ground layer was composed of calcite and traces of quartz. The fine ground layer used under the pigments directly was composed of calcite only. Carbon black was used as black pigment while lead oxide as red pigment, showing the influence of Roman and Greek pigments on Egyptian art in these later periods. Blue colorant was identified as cuprorivaite and yellow pigment was goethite. Animal glue was used in the four pigments as medium colored.

EDXRF analysis of the Pentecost Triptych from the "São Pedro de Miragaia" church (Porto, Portugal)

NASA Astrophysics Data System (ADS)

Roldán, Clodoaldo; Ferrero, José; Juanes, David; Carballo, Jorgelina; Calvo, Ana; Aguiar, Maria

2013-01-01

EDXRF spectrometry was used to determine the pigment composition of the Pentecost Triptych, a Flemish masterpiece located at the Church of São Pedro de Miragaia (Porto, Portugal). The triptych was restored at the beginning of the 20th century and presents a great number of zones with repaints and colour alterations that can be observed on the whole of their surface. The identification of the original pigments and restoration pigments was based on their characteristic colour and the correlation between the detected elements and the named "key elements" of ancient pigments (Ca, Fe, Cu, Hg, Pb,…) and modern pigments (Ti, Ba, Cr, Zn, Co, Cd,…). The analysis of the triptych has been carried out in situ by means of portable instrumentation.

Singlet Molecular Oxygen Generation by Light-Activated DHN-Melanin of the Fungal Pathogen Mycosphaerella fijiensis in Black Sigatoka Disease of Bananas

PubMed Central

Beltrán-García, Miguel J.; Prado, Fernanda M.; Oliveira, Marilene S.; Ortiz-Mendoza, David; Scalfo, Alexsandra C.; Pessoa, Adalberto; Medeiros, Marisa H. G.; White, James F.; Di Mascio, Paolo

2014-01-01

In pathogenic fungi, melanin contributes to virulence, allowing tissue invasion and inactivation of the plant defence system, but has never been implicated as a factor for host cell death, or as a light-activated phytotoxin. Our research shows that melanin synthesized by the fungal banana pathogen Mycosphaerella fijiensis acts as a virulence factor through the photogeneration of singlet molecular oxygen O2 (1Δg). Using analytical tools, including elemental analysis, ultraviolet/infrared absorption spectrophometry and MALDI-TOF mass spectrometry analysis, we characterized both pigment content in mycelia and secreted to the culture media as 1,8-dihydroxynaphthalene (DHN)-melanin type compound. This is sole melanin-type in M. fijiensis. Isolated melanins irradiated with a Nd:YAG laser at 532 nm produced monomol light emission at 1270 nm, confirming generation of O2 (1Δg), a highly reactive oxygen specie (ROS) that causes cellular death by reacting with all cellular macromolecules. Intermediary polyketides accumulated in culture media by using tricyclazole and pyroquilon (two inhibitors of DHN-melanin synthesis) were identified by ESI-HPLC-MS/MS. Additionally, irradiation at 532 nm of that mixture of compounds and whole melanized mycelium also generated O2 (1Δg). A pigmented-strain generated more O2 (1Δg) than a strain with low melanin content. Banana leaves of cultivar Cavendish, naturally infected with different stages of black Sigatoka disease, were collected from field. Direct staining of the naturally infected leaf tissues showed the presence of melanin that was positively correlated to the disease stage. We also found hydrogen peroxide (H2O2) but we cannot distinguish the source. Our results suggest that O2 (1Δg) photogenerated by DHN-melanin may be involved in the destructive effects of Mycosphaerella fijiensis on banana leaf tissues. Further studies are needed to fully evaluate contributions of melanin-mediated ROS to microbial pathogenesis. PMID:24646830

High perfomance liquid chromatography fingerprint analysis for quality control of brotowali (Tinospora crispa)

NASA Astrophysics Data System (ADS)

Syarifah, V. B.; Rafi, M.; Wahyuni, W. T.

2017-05-01

Brotowali (Tinospora crispa) is widely used in Indonesia as ingredient of herbal medicine formulation. To ensure the quality, safety, and efficacy of herbal medicine products, its chemical constituents should be continuously evaluated. High performance liquid chromatography (HPLC) fingerprint is one of powerful technique for this quality control process. In this study, HPLC fingerprint analysis method was developed for quality control of brotowali. HPLC analysis was performed in C18 column and detection was performed using photodiode array detector. The optimum mobile phase for brotowali fingerprint was acetonitrile (ACN) and 0.1% formic acid in gradient elution mode at a flow rate of 1 mL/min. The number of peaks detected in HPLC fingerprint of brotowali was 32 peaks and 23 peaks for stems and leaves, respectively. Berberine as marker compound was detected at retention time of 20.525 minutes. Evaluation of analytical performance including precision, reproducibility, and stability prove that this HPLC fingerprint analysis was reliable and could be applied for quality control of brotowali.

Multielemental speciation analysis by advanced hyphenated technique - HPLC/ICP-MS: A review.

PubMed

Marcinkowska, Monika; Barałkiewicz, Danuta

2016-12-01

Speciation analysis has become an invaluable tool in human health risk assessment, environmental monitoring or food quality control. Another step is to develop reliable multielemental speciation methodologies, to reduce costs, waste and time needed for the analysis. Separation and detection of species of several elements in a single analytical run can be accomplished by high performance liquid chromatography hyphenated to inductively coupled plasma mass spectrometry (HPLC/ICP-MS). Our review assembles articles concerning multielemental speciation determination of: As, Se, Cr, Sb, I, Br, Pb, Hg, V, Mo, Te, Tl, Cd and W in environmental, biological, food and clinical samples analyzed with HPLC/ICP-MS. It addresses the procedures in terms of following issues: sample collection and pretreatment, selection of optimal conditions for elements species separation by HPLC and determination using ICP-MS as well as metrological approach. The presented work is the first review article concerning multielemental speciation analysis by advanced hyphenated technique HPLC/ICP-MS. Copyright © 2016 Elsevier B.V. All rights reserved.

Analysis of biodiesel by high performance liquid chromatography using refractive index detector.

PubMed

Syed, Mahin Basha

2017-01-01

High-performance liquid chromatography (HPLC) was used for the determination of compounds occurring during the production of biodiesel from karanja and jatropha oil. Methanol was used for fast monitoring of conversion of karanja and jatropha oil triacylglycerols to fatty acid methyl esters and for quantitation of residual triacylglycerols (TGs), in the final biodiesel product. The individual sample compounds were identified using HPLC. Analysis of fatty acid methyl esters (FAMES) in blends of biodiesel by HPLC using a refractive index and a UV detector at 238 nm. Individual triacylglycerols, diacylglycerols, monoacylglycerols and methyl esters of oleic, linoleic and linolenic acids and free fatty acids were separated within 40 min. Hence HPLC was found to be best for the analysis of biodiesel. Analysis of biodiesel by HPLC using RID detector. Estimation of amount of FAMES in biodiesel. Individual triacylglycerols, diacylglycerols, monoacylglycerols and methyl esters of oleic, linoleic and linolenic acids and free fatty acids were separated within 40 min.

A multi-analytical investigation of semi-conductor pigments with time-resolved spectroscopy and imaging

NASA Astrophysics Data System (ADS)

Nevin, A.; Cesaratto, A.; D'Andrea, C.; Valentini, Gianluca; Comelli, D.

2013-05-01

We present the non-invasive study of historical and modern Zn- and Cd-based pigments with time-resolved fluorescence spectroscopy, fluorescence multispectral imaging and fluorescence lifetime imaging (FLIM). Zinc oxide and Zinc sulphide are semiconductors which have been used as white pigments in paintings, and the luminescence of these pigments from trapped states is strongly dependent on the presence of impurities and crystal defects. Cadmium sulphoselenide pigments vary in hue from yellow to deep red based on their composition, and are another class of semiconductor pigments which emit both in the visible and the near infrared. The Fluorescence lifetime of historical and modern pigments has been measured using both an Optical Multichannel Analyser (OMA) coupled with a Nd:YAG nslaser, and a streak camera coupled with a ps-laser for spectrally-resolved fluorescence lifetime measurements. For Znbased pigments we have also employed Fluorescence Lifetime Imaging (FLIM) for the measurement of luminescence. A case study of FLIM applied to the analysis of the painting by Vincent Van Gogh on paper - "Les Bretonnes et le pardon de Pont-Aven" (1888) is presented. Through the integration of complementary, portable and non-invasive spectroscopic techniques, new insights into the optical properties of Zn- and Cd-based pigments have been gained which will inform future analysis of late 19th] and early 20th C. paintings.

UPLC-MSE Profiling of Phytoplankton Metabolites: Application to the Identification of Pigments and Structural Analysis of Metabolites in Porphyridium purpureum

PubMed Central

Juin, Camille; Bonnet, Antoine; Nicolau, Elodie; Bérard, Jean-Baptiste; Devillers, Romain; Thiéry, Valérie; Cadoret, Jean-Paul; Picot, Laurent

2015-01-01

A fast and high-resolution UPLC-MSE analysis was used to identify phytoplankton pigments in an ethanol extract of Porphyridium purpureum (Pp) devoid of phycobiliproteins. In a first step, 22 standard pigments were analyzed by UPLC-MSE to build a database including retention time and accurate masses of parent and fragment ions. Using this database, seven pigments or derivatives previously reported in Pp were unequivocally identified: β,β-carotene, chlorophyll a, zeaxanthin, chlorophyllide a, pheophorbide a, pheophytin a, and cryptoxanthin. Minor amounts of Divinyl chlorophyll a, a chemotaxonomic pigment marker for prochlorophytes, were also unequivocally identified using the database. Additional analysis of ionization and fragmentation patterns indicated the presence of ions that could correspond to hydroxylated derivatives of chlorophyll a and pheophytin a, produced during the ethanolic extraction, as well as previously described galactosyldiacylglycerols, the thylakoid coenzyme plastoquinone, and gracilamide B, a molecule previously reported in the red seaweed Gracillaria asiatica. These data point to UPLC-MSE as an efficient technique to identify phytoplankton pigments for which standards are available, and demonstrate its major interest as a complementary method for the structural elucidation of ionizable marine molecules. PMID:25913708

On the discovery of an unusual luminescent pigment in Van Gogh's painting "Les bretonnes et le pardon de pont Aven"

NASA Astrophysics Data System (ADS)

Comelli, D.; Nevin, A.; Brambilla, A.; Osticioli, I.; Valentini, G.; Toniolo, L.; Fratelli, M.; Cubeddu, R.

2012-01-01

Spectrally and lifetime-resolved UV-induced luminescence imaging have been applied to the in situ analysis of Vincent Van Gogh's painting on paper "Les bretonnes et le pardon de pont Aven". The in situ investigation revealed the presence of an unusual microsecond green emission from the white painted areas of the watercolour, and this long-lived emission has not been reported in modern paintings by other authors. In order to attribute and suggest an identification of the luminescent pigment, our analysis was integrated with further measurements on commercially available modern white pigments (zinc white, Lithopone white, Blanc Fixe, zinc sulphide, and copper-doped zinc sulphide) and with an in-depth study of the synthesis processes of zinc-based pigments available at the end of the 19th century. Analysis suggested an attribution of the unusual emission from the Van Gogh painting to a particular variety of zinc sulphide pigment, characterized by the presence of copper impurities which were unintentionally introduced in the crystal matrix during its production. The present study is the first application of lifetime imaging for the mapping of long-lived luminescence from the semi-conductor pigment.

[Immuno-affinity chromatographic purification: the study of methods to test citrinin in monascus products by high performance liquid chromatography].

PubMed

Qiu, Wen-qian; Liu, Xiao-xia; Zheng, Kui-cheng; Fu, Wu-sheng

2012-08-01

To establish a method to test citrinin (CIT) in monascus products by immuno-affinity chromatography (IAC)-high performance liquid chromatography (HPLC), and to detect the content of CIT in monascus products in Fujian province. IAC-HPLC was applied to detect the CIT content in monascus products. The conditions to use HPLC were as follows: C(18) reversed-phase chromatographic column, 150.0 mm×4.6mm×3 µm; mobile phase: the volume ratio of acetonitrile and 0.1% phosphoric acid solution at 65:35; isocratic elution; column temperature: 28°C; flow velocity: 0.8 ml/min; fluorescence detector, excitation wavelength (λ(ex)) was 331 nm and emission wavelength (λ(em)) was 500 nm. The standard curved was established by the linear regression of peak area (Y) to CIT content (X, ng/ml). The accuracy and precision of the method would then be verified. And 32 kinds of monascus products were determined and their color values were compared by this method. The standard curve established in this study was Y = 4634.8X-136.42, r = 1.000; whose limits of detection was 20 µg/kg and the limits of qualification was 64 µg/kg. In the range between 200 and 800 µg/kg, the standard recovery rate was 98.9% - 110.0% (n = 3), and the relative standard deviation (RSD) was 0.51% - 1.76%. Out of the 32 samples, CIT was detected from 11 samples of monascus rice, 9 samples of monascus powder and 5 samples of monascus pigments, the content was around 0.212 - 14.500 mg/kg. 4 out of 7 functional monascus samples were detected out CIT, whose content at 0.142 - 0.275 mg/kg. The method to detect CIT in monascus products by IAC-HPLC has been established.

Study on the Spectral Mixing Model for Mineral Pigments Based on Derivative of Ratio Spectroscopy-Take Vermilion and Stone Yellow for Example

NASA Astrophysics Data System (ADS)

Zhao, H.; Hao, Y.; Liu, X.; Hou, M.; Zhao, X.

2018-04-01

Hyperspectral remote sensing is a completely non-invasive technology for measurement of cultural relics, and has been successfully applied in identification and analysis of pigments of Chinese historical paintings. Although the phenomenon of mixing pigments is very usual in Chinese historical paintings, the quantitative analysis of the mixing pigments in the ancient paintings is still unsolved. In this research, we took two typical mineral pigments, vermilion and stone yellow as example, made precisely mixed samples using these two kinds of pigments, and measured their spectra in the laboratory. For the mixing spectra, both fully constrained least square (FCLS) method and derivative of ratio spectroscopy (DRS) were performed. Experimental results showed that the mixing spectra of vermilion and stone yellow had strong nonlinear mixing characteristics, but at some bands linear unmixing could also achieve satisfactory results. DRS using strong linear bands can reach much higher accuracy than that of FCLS using full bands.

Identification of the anti-oxidant components in a two-step solvent extract of bovine bile lipid: Application of reverse phase HPLC, mass spectrometry and fluorimetric assays.

PubMed

Singh, Namrata; Bhattacharyya, Debasish

2016-04-15

An ether extract of nine different bacterial metabolites in combination with two solvent extract (ether followed by ethanol) of bile lipids from ox gall bladder is used as an immune stimulator drug. Over the years bile acids are discussed regarding their anti-oxidant and lipid peroxidation properties. Since some of the bile acids are known to be potent antioxidants, presence of similar activity in the solvent extract of ox bile lipid was investigated using TLC and reverse phase HPLC systems. Fractions from HPLC were analyzed with mass spectrometry using electrospray ionization. The presence of twelve different bile acids along with other substances in small proportions including fatty acids, sulfate conjugates and bile pigments were confirmed. The twelve separated peaks had similar retention times as those of tauroursodeoxycholic acid, glycoursodeoxycholic acid, taurocholic acid, glycocholic acid, glycochenodeoxycholic acid, taurochenodeoxycholic acid, taurodeoxycholic acid, cholic acid, ursodeoxycholic acid, chenodeoxycholic acid, deoxycholic acid, and lithocholic acid. Subsequently, all fractions were tested for their anti-oxidative property on HepG2 cells exposed to H2O2 that served as an oxidative injury model. Four fluorescent dyes H2DCF DA, MitoSOX red, Amplex red and DAF-2 DA were used for estimation of reactive radicals in the HepG2 cells. Among the separated bile acids, tauroursodeoxycholic acid, glycoursodeoxycholic acid and ursodeoxycholic acid prevented the HepG2 cells from H2O2-induced oxidative stress. Copyright © 2015 Elsevier B.V. All rights reserved.

Elucidating the Relationship Between Phytoplankton and Primary Production in the Sargasso Sea Using New Observations of Nanoplankton and Picoplankton.

NASA Astrophysics Data System (ADS)

Matheson, J.; Johnson, R. J.; Bates, N. R.; Parsons, R. J.

2016-02-01

Attempts to model primary production in the subsurface of the Sargasso Sea frequently use HPLC marker pigments to infer phytoplankton community structure, which relies upon assumptions about the phytoplankton community typically determined with limited site-specific data. Recent estimates suggest that nano- and picoplankton account for 90% of the phytoplankton community at BATS and factors such as elevated growth rates and high abundances likely allow these two size classes to exert a strong influence on primary production. To help assess the contribution of nano- and picoplankton on primary production at the BATS site we determine abundances and biovolumes through direct measurements with epifluorescence microscopy in conjunction with flow cytometer picoplankton counts. Using this approach we are able to quantify prymnesiophytes, heterotrophic nano- and dinoflagellates, mixotrophic dinoflagellates, ciliates, diatoms, pico- and nano eukaryotes, and Prochlorococcus. Preliminary analysis of summertime distributions show prymnesiophytes are the dominant nanoplankton group (average upper 140 m concentration of 500 cells ml-1) although heterotrophic nano- and dinoflagellates makeup a greater fraction of nanoplankton biovolume. During the summer period, pico-eukaryotes and Prochlorococcus were found to be the dominant picoplankton groups, which both increased with depth down to the deep chlorophyll maximum where they appear to drive variability. Using these direct observations we investigate the seasonal relationship between phytoplankton community and primary production, specifically by contrasting the stratified summer phase with a well-mixed winter system. Finally, we use these community structure observations with HPLC data to develop algorithms for taxonomy models (i.e. CHEMTAX) to assess modes of variability in phytoplankton community and consequential influences on primary production for the past 25 years at the BATS site.

Phenolic Profiling and Antioxidant Capacity of Eugenia uniflora L. (Pitanga) Samples Collected in Different Uruguayan Locations

PubMed Central

Baenas, Nieves; Gironés-Vilaplana, Amadeo; Cesio, María Verónica; Heinzen, Horacio

2018-01-01

The use of nutrient-rich foods to enhance the wellness, health and lifestyle habits of consumers is globally encouraged. Native fruits are of great interest as they are grown and consumed locally and take part of the ethnobotanic knowledge of the population. Pitanga is an example of a native fruit from Uruguay, consumed as a jelly or an alcoholic beverage. Pitanga has a red-violet pigmentation, which is a common trait for foods that are a good source of antioxidants. Hence, fruits from different Uruguayan regions were analyzed via miniaturized sample preparation method, HPLC-DAD-ESI/MSn and RP-HPLC-DAD techniques to identify and quantify phenolic compounds, respectively. The antioxidant capacity was evaluated via DPPH and ORAC (Oxygen Radical Absorbance Capacity) assays. A multivariate linear regression was applied to correlate the observed antioxidant capacity with the phenolic content. Furthermore, Principal Components Analysis was performed to highlight characteristics between the various samples studied. The main results indicated differences between northern and southern Uruguayan samples. Delphinidin-3-hexoside was present in southern samples (mean of 293.16 µmol/100 g dry weight (DW)) and absent in the sample collected in the north (sample 3). All the samples contain high levels of cyanidin-3-hexoside, but a noticeable difference was found between the northern sample (150.45 µmol/100 g DW) and the southern sample (1121.98 µmol/100 g DW). The antioxidant capacity (mean ORAC of 56370 µmol Trolox®/100 g DW) were high in all the samples compared to the Food and Drug Administration (FDA) database of similar berry-fruits. The results of this study highlight the nutraceutical value of a native fruit that has not been exploited until now. PMID:29695116

Phenolic Profiling and Antioxidant Capacity of Eugenia uniflora L. (Pitanga) Samples Collected in Different Uruguayan Locations.

PubMed

Migues, Ignacio; Baenas, Nieves; Gironés-Vilaplana, Amadeo; Cesio, María Verónica; Heinzen, Horacio; Moreno, Diego A

2018-04-24

The use of nutrient-rich foods to enhance the wellness, health and lifestyle habits of consumers is globally encouraged. Native fruits are of great interest as they are grown and consumed locally and take part of the ethnobotanic knowledge of the population. Pitanga is an example of a native fruit from Uruguay, consumed as a jelly or an alcoholic beverage. Pitanga has a red-violet pigmentation, which is a common trait for foods that are a good source of antioxidants. Hence, fruits from different Uruguayan regions were analyzed via miniaturized sample preparation method, HPLC-DAD-ESI/MS n and RP-HPLC-DAD techniques to identify and quantify phenolic compounds, respectively. The antioxidant capacity was evaluated via DPPH and ORAC (Oxygen Radical Absorbance Capacity) assays. A multivariate linear regression was applied to correlate the observed antioxidant capacity with the phenolic content. Furthermore, Principal Components Analysis was performed to highlight characteristics between the various samples studied. The main results indicated differences between northern and southern Uruguayan samples. Delphinidin-3-hexoside was present in southern samples (mean of 293.16 µmol/100 g dry weight (DW)) and absent in the sample collected in the north (sample 3). All the samples contain high levels of cyanidin-3-hexoside, but a noticeable difference was found between the northern sample (150.45 µmol/100 g DW) and the southern sample (1121.98 µmol/100 g DW). The antioxidant capacity (mean ORAC of 56370 µmol Trolox ® /100 g DW) were high in all the samples compared to the Food and Drug Administration (FDA) database of similar berry-fruits. The results of this study highlight the nutraceutical value of a native fruit that has not been exploited until now.

A flavonoid from Brassica rapa flower as the UV-absorbing nectar guide.

PubMed

Sasaki, Katsunori; Takahashi, Takashi

2002-10-01

The corolla of Brassica rapa has an UV-absorbing zone in its center, known as the nectar guide for attracting pollinating insects. The pigment which plays the role of the nectar guide was isolated from the petals and identified to be isorhamnetin 3,7-O-di-beta-D-glucopyranoside on the basis of MS and NMR spectroscopic data. The D-, L-configurations of the sugar moieties were determined by the fluorometric HPLC method. In plants raised in open field, there was a 13-fold higher content of the compound in the basal parts of the petals compared with the apical parts. This difference in flavonoid content is presumed to contribute to the visual attractiveness of B. rapa flowers to insect pollinators.

Transcriptomic analysis of skin pigmentation variation in the Virginia opossum (Didelphis virginiana).

PubMed

Nigenda-Morales, Sergio F; Hu, Yibo; Beasley, James C; Ruiz-Piña, Hugo A; Valenzuela-Galván, David; Wayne, Robert K

2018-06-01

Skin pigmentation and coat pigmentation are two of the best-studied examples of traits under natural selection given their quantifiable fitness interactions with the environment (e.g., camouflage) and signalling with other organisms (e.g., warning coloration). Previous morphological studies have found that skin pigmentation variation in the Virginia opossum (Didelphis virginiana) is associated with variation in precipitation and temperatures across its distribution range following Gloger's rule (lighter pigmentation in temperate environments). To investigate the molecular mechanism associated with skin pigmentation variation, we used RNA-Seq and quantified gene expression of wild opossums from tropical and temperate populations. Using differential expression analysis and a co-expression network approach, we found that expression variation in genes with melanocytic and immune functions is significantly associated with the degree of skin pigmentation variation and may be underlying this phenotypic difference. We also found evidence suggesting that the Wnt/β-catenin signalling pathway might be regulating the depigmentation observed in temperate populations. Based on our study results, we present several alternative hypotheses that may explain Gloger's rule pattern of skin pigmentation variation in opossum, including changes in pathogen diversity supporting a pathogen-resistant hypothesis, thermal stress associated with temperate environments, and pleiotropic and epistatic interactions between melanocytic and immune genes. © 2018 John Wiley & Sons Ltd.

Traceability of sulfonamide antibiotic treatment by immunochemical analysis of farm animal hair samples.

PubMed

Adrian, Javier; Gratacós-Cubarsí, Marta; Sánchez-Baeza, Francisco; Garcia Regueiro, Jose-Antonio; Castellari, Massimo; Marco, M-Pilar

2009-10-01

The use of hair to trace use of unauthorized substances, therapeutic agents, or their misuse is becoming very attractive since residues can be detected for a long time after treatment. For this purpose, an indirect enzyme-linked immunosorbent assay (ELISA) has been evaluated for its capability to trace sulfonamide antibiotic treatment by analyzing cattle and pig hair samples. Pigmented and nonpigmented hair samples from control and sulfamethazine (SMZ)-treated pigs and calves were collected, extracted under different alkaline conditions, and analyzed by ELISA after just diluting the extracts with the assay buffer. Data analysis following the European recommendations for screening methods demonstrates that the ELISA can detect SMZ in hair samples with a limit of detection (90% of the zero dose (IC(90))) between 30 and 75 ng g(-1). The same samples have been analyzed by HPLC after a dual solid-phase extraction. The ELISA results matched very well those obtained by the chromatographic method, demonstrating that the immunochemical method can be used as a screening tool to trace animal treatments. Between the benefits of this method are the possibility to directly analyze hair extracts with sufficient detectability and its high-throughput capability. Preliminary validation data are reported using an experimental approach inspired on the Commission Decision 2002/657/EC criteria for screening methods.

[A new HPLC procedure for cyclamate in food with pre-chromatographic derivatization].

PubMed

Schwedt, G; Hauck, M

1988-08-01

A high-pressure liquid chromatography (HPLC) procedure for the detection of cyclamate in liquid and solid samples is presented, which depends on oxidation and the reaction of cyclohexylamine with o-phthaldialdehyde to form a condensation product. The results of the HPLC analysis, using an RP-C 18 separation system with UV detection at 242 nm are reported. Contents, from 2 to 400 mg/l, can be detected in less than 2 h (HPLC analysis within 20 min) with relative standard deviations of 4%. Only for cucumber infusions were incomplete recoveries of 68% obtained.

High lenticular tolerance to ultraviolet radiation-B by pigmented guinea-pig; application of a safety limit strategy for UVR-induced cataract.

PubMed

Mody, Vino C; Kakar, Manoj; Söderberg, Per G; Löfgren, Stefan

2012-05-01

The purpose of this study was to determine a threshold measure, maximum tolerable dose (MTD), for avoidance of UVR-B-induced cataract in the pigmented guinea-pig. Thirty pupil-dilated anesthetized young female guinea-pigs, divided into five equal groups, received between 0 and 84.9 kJ/m(2) unilateral UVR-B. Lens extraction and in vitro lens photography occurred 24 hr after exposure. Measurement of intensity of lens light scattering served as quantifying tool for the degree of cataract. Data analysis included regression, using a second order polynomial model. The applied MTD concept was based on the UVR-B dose-response curve obtained for the pigmented guinea-pig. A smaller number of pigmented guinea-pigs, pigmented rats and albino rats underwent morphometric analysis of the anterior segment geometry. All eyes exposed to UVR-B developed cataract in the anterior subcapsular region. MTD for avoidance of UVR-B-induced cataract was 69.0 kJ/m(2) in the pigmented guinea-pig. Iris was considerably thicker in the guinea-pig than in the rats. Lens blockage by the dilated iris was lowest in the guinea-pig. Maximum tolerable dose for avoidance of UVR-B-induced cataract in the pigmented guinea-pig was 69.0 kJ/m(2), over 10-fold higher than the threshold 5 kJ/m(2) obtained by Pitts et al. in the pigmented rabbit. Maximum tolerable dose is an appropriate method for estimation of toxicity for UVR-B-induced cataract in the guinea-pig. The pigmented guinea-pig is significantly less sensitive to UVR-B exposure than the pigmented rabbit and pigmented rat. © 2010 The Authors. Journal compilation © 2010 Acta Ophthalmol.

High performance thin layer chromatography (HPTLC) and high performance liquid chromatography (HPLC) for the qualitative and quantitative analysis of Calendula officinalis-advantages and limitations.

PubMed

Loescher, Christine M; Morton, David W; Razic, Slavica; Agatonovic-Kustrin, Snezana

2014-09-01

Chromatography techniques such as HPTLC and HPLC are commonly used to produce a chemical fingerprint of a plant to allow identification and quantify the main constituents within the plant. The aims of this study were to compare HPTLC and HPLC, for qualitative and quantitative analysis of the major constituents of Calendula officinalis and to investigate the effect of different extraction techniques on the C. officinalis extract composition from different parts of the plant. The results found HPTLC to be effective for qualitative analysis, however, HPLC was found to be more accurate for quantitative analysis. A combination of the two methods may be useful in a quality control setting as it would allow rapid qualitative analysis of herbal material while maintaining accurate quantification of extract composition. Copyright © 2014 Elsevier B.V. All rights reserved.

[The qualitative analysis and quantitative analysis of purification of salvianolic acids by macroreticular resin].

PubMed

Fang, Xin-sheng; Tan, Xiao-mei

2005-09-01

To purify salvianolic acids by macroreticular resin,then mensurate the contents of salvianolic acids and analyse the chromatogram with HPLC. Make salvianolic acids with macroreticular resin; mensurate the content of Salvianolic acids with UV spestrophotometry: the control compound is protocaechuic aldehyde, and the wavelength is 281 nm. Analysis the chromatogram with HPLC, and compare the chromatogram in different technics: zorbax ODS column (4.6 mm x 250 mm, 5 microm), mobilephase: 1% aceticacid-water and methanol in different proportions, the wavelength is 281 nm. The contents of salvianolic acids is 53.8%; HPLC chromatogram indicate that the method is reasonable to make salvianolic acids. Determination of contents and HPLC chromatogram can control the quality of Salvianolic acids more accurately.

A Short History of the Chemistry of Painting.

ERIC Educational Resources Information Center

Friedstein, Harriet G.

1981-01-01

Includes information on: (1) relationship of art and science; (2) paintings' early history; (3) Egyptian, Greek, Chinese, Byzantine, and Medieval painting; (4) chemical analysis of pigments; (5) chemistry of early pigments; and (6) paint media. Tabular data are provided on chemical names for artists' pigments with their earliest known dates. (CS)

The effect of re-dissolution solvents and HPLC columns on the analysis of mycosporine-like amino acids in the eulittoral macroalgae Prasiola crispa and Porphyra umbilicalis

NASA Astrophysics Data System (ADS)

Karsten, Ulf; Escoubeyrou, Karine; Charles, François

2009-09-01

Many macroalgal species that are regularly exposed to high solar radiation such as the eulittoral green alga Prasiola crispa and the red alga Porphyra umbilicalis synthesize and accumulate high concentrations of mycosporine-like amino acids (MAAs) as UV-sunscreen compounds. These substances are typically extracted with a widely used standard protocol following quantification by various high performance liquid chromatography (HPLC) techniques. However, further preparation steps prior to HPLC analysis as well as different HPLC column types have not been systematically checked regarding separation quality and reproducibility. Therefore pure methanol, distilled water and HPLC eluent were evaluated as re-dissolution solvent for dried Prasiola and Porphyra extracts, which were subsequently analyzed on three reversed-phase C8 and C18 HPLC columns. The data indicate that distilled water and the HPLC eluent gave almost identical peak patterns and MAA contents on the C8 and C18 columns. In contrast, the application of the widely used methanol led to double peaks or even the loss of specific peaks as well as to a strong decline in total MAA amounts ranging from about 35% of the maximum in P. crispa to 80% of the maximum in P. umbilicalis. Consequently, methanol should be avoided as re-dissolution solvent for the HPLC sample preparation. An improved protocol for the MAA analysis in macroalgae in combination with a reliable C18 column is suggested.

HPLC-Orbitrap analysis for identification of organic molecules in complex material

NASA Astrophysics Data System (ADS)

Gautier, T.; Schmitz-Afonso, I.; Carrasco, N.; Touboul, D.; Szopa, C.; Buch, A.; Pernot, P.

2015-10-01

We performed High Performance Liquid Chromatography (HPLC) coupled to Orbitrap High Resolution Mass Spectrometry (OHR MS) analysis of Titan's tholins. This analysis allowed us to determine the exact composition and structure of some of the major components of tholins.

Raman spectroscopy for the identification of pigments and color measurement in Dugès watercolors

NASA Astrophysics Data System (ADS)

Frausto-Reyes, C.; Ortiz-Morales, M.; Bujdud-Pérez, J. M.; Magaña-Cota, G. E.; Mejía-Falcón, R.

2009-12-01

Spectroscopic and colorimetric analysis of a representative set of Dugès watercolor paintings was performed. These paintings were the result of scientific studies carried out by the zoologist Alfredo Dugès, who recorded the fauna of the Mexican Republic between 1853 and 1910. Micro-Raman spectroscopy, with an excitation wavelength of 830 nm, and colorimetric techniques were employed in order to understand if different colors with the same hue were reproduced using the same pigments. The color coordinates of the measured areas were obtained in the CIE L* a* b* color space. Raman analysis showed that, in some cases, to reproduce colors with the same hue the pigment employed was not the same. Pigments identified in the watercolors were vermilion, carbon-based black, lead white, gamboge and chrome yellow, Prussian and ultramarine blue. Some of these pigments have been used since ancient times, others as Prussian blue, chrome yellow and synthetic ultramarine blue arrived to the market at the beginning of the 18th and 19th centuries, respectively. Furthermore, regarding the white color, instead of left the paper unpainted, lead white was detected in the eye of a bird. The green color was obtained by mixing Prussian blue with chrome yellow. The results of this work show the suitability of using Raman spectroscopy for watercolor pigment analysis and colorimetric techniques to measure the color of small areas (246 μm × 246 μm) that was the case for the lead white pigment.

Chromatographic and Spectrophotometric Analysis of Phenolic Compounds from Fruits of Libidibia ferrea Martius

PubMed Central

Ferreira, Magda R. A.; Fernandes, Mônica T. M.; da Silva, Wliana A. V.; Bezerra, Isabelle C. F.; de Souza, Tatiane P.; Pimentel, Maria F.; Soares, Luiz A. L.

2016-01-01

Background: Libidibia ferrea (Mart. ex Tul.) L.P. Queiroz (Fabaceae) is a tree which is native to Brazil, widely known as “Jucá,” where its herbal derivatives are used in folk medicine with several therapeutic properties. The constituents, which have already been described in the fruit, are mainly hydrolysable tannins (gallic acid [GA] and ellagic acid [EA]). Objective: The aim of this study was to investigate the phenolic variability in the fruit of L. ferrea by ultraviolet/visible (UV/VIS) and chromatographic methods (high-performance liquid chromatography [HPLC]/high-performance thin layer chromatography [HPTLC]). Materials and Methods: Several samples were collected from different regions of Brazil and the qualitative (fingerprints by HPTLC and HPLC) and quantitative analysis (UV/VIS and HPLC) of polyphenols were performed. Results: The HPTLC and HPLC profiles allowed separation and identification of both major analytical markers: EA and GA. The chemical profiles were similar in a number of spots or peaks for the samples, but some differences could be observed in the intensity or area of the analytical markers for HPTLC or HPLC, respectively. Regarding the quantitative analysis, the polyphenolic content by UV/VIS ranged from 13.99 to 37.86 g% expressed as GA or from 10.75 to 29.09 g% expressed as EA. The contents of EA and GA by liquid chromatography-reversed phase (LC-RP) method ranged from 0.57 to 2.68 g% and from 0.54 to 3.23 g%, respectively. Conclusion: The chemical profiles obtained by HPTLC or HPLC, as well as the quantitative analysis by spectrophotometry or LC-RP method, were suitable for discrimination of each herbal sample and can be used as tools for the comparative analysis of the fruits from L. ferrea. SUMMARY The polyphenols of fruits of Libidibia ferrea can be quantified by UV/VIS and HPLCThe HPLC method was able to detect the gallic and ellagic acids in several samples of fruits of Libidibia ferreaThe phenolic profiles of fruits from Libidibia ferrea by HPTLC and HPLC were reproductible. Abbreviations used: HPTLC: high performance thin layer chromatography, HPLC: high performance liquid chromatography, UV-Vis: spectrophotometry PMID:27279721

Interface for liquid chromatograph-mass spectrometer

DOEpatents

Andresen, B.D.; Fought, E.R.

1989-09-19

A moving belt interface is described for real-time, high-performance liquid chromatograph (HPLC)/mass spectrometer (MS) analysis which strips away the HPLC solvent as it emerges from the end of the HPLC column and leaves a residue suitable for mass-spectral analysis. The interface includes a portable, stand-alone apparatus having a plural stage vacuum station, a continuous ribbon or belt, a drive train magnetically coupled to an external drive motor, a calibrated HPLC delivery system, a heated probe tip and means located adjacent the probe tip for direct ionization of the residue on the belt. The interface is also capable of being readily adapted to fit any mass spectrometer. 8 figs.

Interface for liquid chromatograph-mass spectrometer

DOEpatents

Andresen, Brian D.; Fought, Eric R.

1989-01-01

A moving belt interface for real-time, high-performance liquid chromatograph (HPLC)/mass spectrometer (MS) analysis which strips away the HPLC solvent as it emerges from the end of the HPLC column and leaves a residue suitable for mass-spectral analysis. The interface includes a portable, stand-alone apparatus having a plural stage vacuum station, a continuous ribbon or belt, a drive train magnetically coupled to an external drive motor, a calibrated HPLC delivery system, a heated probe tip and means located adjacent the probe tip for direct ionization of the residue on the belt. The interface is also capable of being readily adapted to fit any mass spectrometer.

Production of colourful pigments consisting of amorphous arrays of silica particles.

PubMed

Yoshioka, Shinya; Takeoka, Yukikazu

2014-08-04

It is desirable to produce colourful pigments that have anti-fading properties and are environmentally friendly. In this Concept, we describe recently developed pigments that exhibit such characteristics. The pigments consist of amorphous arrays of submicron silica particles, and they exhibit saturated and angle-independent structural colours. Variously coloured pigments can be produced by changing the size of the particles, and the saturation of the colour can be controlled by incorporating small amounts of black particles. We review a simple analysis that is useful for interpreting the angular independence of the structural colours and discuss the remaining tasks that must be accomplished for the realistic application of these pigments. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Adulteration and cultivation region identification of American ginseng using HPLC coupled with multivariate analysis

PubMed Central

Yu, Chunhao; Wang, Chong-Zhi; Zhou, Chun-Jie; Wang, Bin; Han, Lide; Zhang, Chun-Feng; Wu, Xiao-Hui; Yuan, Chun-Su

2014-01-01

American ginseng (Panax quinquefolius) is originally grown in North America. Due to price difference and supply shortage, American ginseng recently has been cultivated in northern China. Further, in the market, some Asian ginsengs are labeled as American ginseng. In this study, forty-three American ginseng samples cultivated in the USA, Canada or China were collected and 14 ginseng saponins were determined using HPLC. HPLC coupled with hierarchical cluster analysis and principal component analysis was developed to identify the species. Subsequently, an HPLC-linear discriminant analysis was established to discriminate cultivation regions of American ginseng. This method was successfully applied to identify the sources of 6 commercial American ginseng samples. Two of them were identified as Asian ginseng, while 4 others were identified as American ginseng, which were cultivated in the USA (3) and China (1). Our newly developed method can be used to identify American ginseng with different cultivation regions. PMID:25044150

Molar absorptivity (ε) and spectral characteristics of cyanidin-based anthocyanins from red cabbage.

PubMed

Ahmadiani, Neda; Robbins, Rebecca J; Collins, Thomas M; Giusti, M Monica

2016-04-15

Red cabbage extract contains mono and di-acylated cyanidin (Cy) anthocyanins and is often used as food colorants. Our objectives were to determine the molar absorptivity (ε) of different red cabbage Cy-derivatives and to evaluate their spectral behaviors in acidified methanol (MeOH) and buffers pH 1-9. Major red cabbage anthocyanins were isolated using a semi-preparatory HPLC, dried and weighed. Pigments were dissolved in MeOH and diluted with either MeOH (0.1% HCl) or buffers to obtain final concentrations between 5×10(-5) and 1×10(-3) mol/L. Spectra were recorded and ε calculated using Lambert-Beer's law. The ε in acidified MeOH and buffer pH 1 ranged between ~16,000-30,000 and ~13,000-26,000 L/mol cm, respectively. Most pigments showed higher ε in pH 8 than pH 2, and lowest ε between pH 4 and 6. There were bathochromic shifts (81-105 nm) from pH 1 to 8 and hypsochromic shifts from pH 8 to 9 (2-19 nm). Anthocyanins molecular structures and the media were important variables which greatly influenced their ε and spectral behaviors. Copyright © 2015 Elsevier Ltd. All rights reserved.

Cellulosic fabrics printing with multifunctional encapsulated phthalocyanine pigment blue using phase separation method.

PubMed

Haroun, Ahmed A; Diab, H A; Hakeim, O A

2016-08-01

Aqueous dispersions of citric-acrylate (CAC) oligomer encapsulating C.I. Pigment Blue 15:3 (PB15:3) in the presence of glutaraldhyde were formulated using the phase separation method. FT-IR spectroscopy and centrifuge sedimentation are performed to confirm the encapsulation of pigment into CAC oligomer. The prepared capsules were characterized using thermal gravimetric analysis (TGA) and transmission electron microscope (TEM). The results revealed that the encapsulated pigment had a profound multifunctional impact and minimized the driving force of pigment printing on the cellulosic fabrics. Besides, the encapsulated pigment accelerated the pigment fixation on cellulosic fabrics without drying in one step and reduced the required amount of the binder, compared with the control sample. Furthermore, the printed fabrics exhibited good antibacterial performance against both Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). The presence of the crosslinker could be stabilized the encapsulated pigment on the cellulosic fabrics. Moreover, the light and washing fastness for the printed fabrics using encapsulated pigment are higher than that in case of using control samples. Copyright © 2016 Elsevier Ltd. All rights reserved.

Iris phenotypes and pigment dispersion caused by genes influencing pigmentation

PubMed Central

Hawes, Norman L.; Trantow, Colleen M.; Chang, Bo; John, Simon W.M.

2010-01-01

Summary Spontaneous mutations altering mouse coat colors have been a classic resource for discovery of numerous molecular pathways. Although often overlooked, the mouse iris is also densely pigmented and easily observed, thus representing a similarly powerful opportunity for studying pigment cell biology. Here, we present an analysis of iris phenotypes among sixteen mouse strains with mutations influencing melanosomes. Many of these strains exhibit biologically and medically relevant phenotypes, including pigment dispersion, a common feature of several human ocular diseases. Pigment dispersion was identified in several strains with mutant alleles known to influence melanosomes, including beige, light, and vitiligo. Pigment dispersion was also detected in the recently arising spontaneous coat color variant, nm2798. We have identified the nm2798 mutation as a missense mutation in the Dct gene, an identical re-occurrence of the slaty light mutation. These results suggest that dysregulated events of melanosomes can be potent contributors to the pigment dispersion phenotype. Combined, these findings illustrate the utility of studying iris phenotypes as a means of discovering new pathways, and re-linking old ones, to processes of pigmented cells in health and disease. PMID:18715234

Iris phenotypes and pigment dispersion caused by genes influencing pigmentation.

PubMed

Anderson, Michael G; Hawes, Norman L; Trantow, Colleen M; Chang, Bo; John, Simon W M

2008-10-01

Spontaneous mutations altering mouse coat colors have been a classic resource for discovery of numerous molecular pathways. Although often overlooked, the mouse iris is also densely pigmented and easily observed, thus representing a similarly powerful opportunity for studying pigment cell biology. Here, we present an analysis of iris phenotypes among 16 mouse strains with mutations influencing melanosomes. Many of these strains exhibit biologically and medically relevant phenotypes, including pigment dispersion, a common feature of several human ocular diseases. Pigment dispersion was identified in several strains with mutant alleles known to influence melanosomes, including beige, light, and vitiligo. Pigment dispersion was also detected in the recently arising spontaneous coat color variant, nm2798. We have identified the nm2798 mutation as a missense mutation in the Dct gene, an identical re-occurrence of the slaty light mutation. These results suggest that dysregulated events of melanosomes can be potent contributors to the pigment dispersion phenotype. Combined, these findings illustrate the utility of studying iris phenotypes as a means of discovering new pathways, and re-linking old ones, to processes of pigmented cells in health and disease.

High Throughput, High Content Screening for Novel Pigmentation Regulators Using a Keratinocyte/Melanocyte Co-culture System

PubMed Central

Lee, Ju Hee; Chen, Hongxiang; Kolev, Vihren; Aull, Katherine H.; Jung, Inhee; Wang, Jun; Miyamoto, Shoko; Hosoi, Junichi; Mandinova, Anna; Fisher, David E.

2014-01-01

Skin pigmentation is a complex process including melanogenesis within melanocytes and melanin transfer to the keratinocytes. To develop a comprehensive screening method for novel pigmentation regulators, we used immortalized melanocytes and keratinocytes in co-culture to screen large numbers of compounds. High-throughput screening plates were subjected to digital automated microscopy to quantify the pigmentation via brightfield microscopy. Compounds with pigment suppression were secondarily tested for their effects on expression of MITF and several pigment regulatory genes, and further validated in terms of non-toxicity to keratinocytes/melanocytes and dose dependent activity. The results demonstrate a high-throughput, high-content screening approach, which is applicable to the analysis of large chemical libraries using a co-culture system. We identified candidate pigmentation inhibitors from 4,000 screened compounds including zoxazolamine, 3-methoxycatechol, and alpha-mangostin, which were also shown to modulate expression of MITF and several key pigmentation factors, and are worthy of further evaluation for potential translation to clinical use. PMID:24438532

Modification of flower colour by suppressing β-ring carotene hydroxylase genes in Oncidium.

PubMed

Wang, H-M; To, K-Y; Lai, H-M; Jeng, S-T

2016-03-01

Oncidium 'Gower Ramsey' (Onc. GR) is a popular cut flower, but its colour is limited to bright yellow. The β-ring carotene hydroxylase (BCH2) gene is involved in carotenoid biogenesis for pigment formation. However, the role of BCH2 in Onc. GR is poorly understood. Here, we investigated the functions of three BCH2 genes, BCH-A2, BCH-B2 and BCH-C2 isolated from Onc. GR, to analyse their roles in flower colour. RT-PCR expression profiling suggested that BCH2 was mainly expressed in flowers. The expression of BCH-B2 remained constant while that of BCH-A2 gradually decreased during flower development. Using Agrobacterium tumefaciens to introduce BCH2 RNA interference (RNAi), we created transgenic Oncidium plants with down-regulated BCH expression. In the transgenic plants, flower colour changed from the bright yellow of the wild type to light and white-yellow. BCH-A2 and BCH-B2 expression levels were significantly reduced in the transgenic flower lips, which make up the major portion of the Oncidium flower. Sectional magnification of the flower lip showed that the amount of pigmentation in the papillate cells of the adaxial epidermis was proportional to the intensity of yellow colouration. HPLC analyses of the carotenoid composition of the transgenic flowers suggested major reductions in neoxanthin and violaxanthin. In conclusion, BCH2 expression regulated the accumulation of yellow pigments in the Oncidium flower, and the down-regulation of BCH-A2 and BCH-B2 changed the flower colour from bright yellow to light and white-yellow. © 2015 German Botanical Society and The Royal Botanical Society of the Netherlands.

Mass Spectrometric Analysis of Synthetic Organic Pigments.

PubMed

Sugaya, Naeko; Takahashi, Mitsuko; Sakurai, Katsumi; Tanaka, Nobuko; Okubo, Ichiro; Kawakami, Tsuyoshi

2018-04-18

Though synthetic organic colorants are used in various applications nowadays, there is the concern that impurities by-produced during the manufacturing and degradation products in some of these colorants are persistent organic pollutants and carcinogens. Thus, it is important to identify the synthetic organic colorants in various products, such as commercial paints, ink, cosmetics, food, textile, and plastics. Dyes, which are soluble in water and other solvents, could be analyzed by chromatographic methods. In contrast, it is difficult to analyze synthetic organic pigments by these methods because of their insolubility. This review is an overview of mass spectrometric analysis of synthetic organic pigments by various ionization methods. We highlight a recent study of textile samples by atmospheric pressure solid analysis probe MS. Furthermore, the mass spectral features of synthetic organic pigments and their separation from other components such as paint media and plasticizers are discussed.

Ultrastructural and biochemical analysis of epidermal xanthophores and dermal chromatophores of the teleost Sparus aurata.

PubMed

Ferrer, C; Solano, F; Zuasti, A

1999-04-01

We have studied the pigmentary system of the teleost Sparus aurata skin by electron microscopy and chromatographic analysis. Under electron microscopy, we found the dermis to contain the three major types of recognized chromatophores: melanophores, xanthophores and iridophores. Melanophores were more abundant in the dorsal region, whereas the iridophores were more abundant in the ventral region. The most important discovery was that of epidermal xanthophores. Epidermal xanthophores were the only chromatophores in the epidermis, something only found in S aurata and in a teleost species living in the Antartic sea. In contrast, the biochemical analysis did not establish any special characteristics: we found pteridine and flavin pigments located mostly in the pigmented dorsal region. Riboflavin and pterin were two of the most abundant coloured pigment types, but other colourless pigments such as xanthopterin and isoxanthopterin were also detected.

Studies on chemical constituents and bioactivity of Rosa micrantha: an alternative antioxidants source for food, pharmaceutical, or cosmetic applications.

PubMed

Guimarães, Rafaela; Barros, Lillian; Carvalho, Ana Maria; Ferreira, Isabel C F R

2010-05-26

Rose species have long been used for food and medicinal purposes. Rosa micrantha is one of the rose species that grow feral in the northeastern Portuguese region so-called Nordeste Transmontano. For the first time, chemical composition and bioactivity of their petals, fertilized flowers, unripe, ripening, and overripe hips were evaluated in order to valorize them as sources of important phytochemicals. Chemical characterization included determination of proteins, fats, ash, and carbohydrates, particularly sugars, by HPLC-RI, fatty acids by GC-FID, tocopherols by HPLC-fluorescence, and phenolics, flavonoids, carotenoids, and ascorbic acid by spectrophotometric techniques. Bioactivity was evaluated through screening of antioxidant properties: radical scavenging effects, reducing power, and inhibition of lipid peroxidation. Ripening and overripe hips showed high nutritional value including proteins, carbohydrates, omega-3 and omega-6 fatty acids, energy, sugars, particularly the reducing sugars fructose and glucose, and ascorbic acid (>693 mg/100 g). Fertilized flowers and petals revealed the highest antioxidant activity (EC(50) > 152 microg/mL) and phenolics, flavonoids, and tocopherols contents (>35 mg/100 g). Furthermore, petals, ripening, and overripe hips are important sources of carotenoid pigments (>64 mg/100 g). Because of the diversity and abundance of antioxidants found in this species, some food, cosmetic, and pharmaceutical applications could be explored.

Carotenoid composition of jackfruit (Artocarpus heterophyllus), determined by HPLC-PDA-MS/MS.

PubMed

de Faria, A F; de Rosso, V V; Mercadante, A Z

2009-06-01

Carotenoids are pigments responsible for the yellow-reddish color of many foods and are related to important functions and physiological actions, preventing several chronic-degenerative diseases. The objective of this study was to confirm the carotenoid composition of jackfruit by high-performance liquid chromatography connected to photodiode array and mass spectrometry detectors (HPLC-PDA-MS/MS). The main carotenoids were all-trans-lutein (24-44%), all-trans-beta-carotene (24-30%), all-trans-neoxanthin (4-19%), 9-cis-neoxanthin (4-9%) and 9-cis-violaxanthin (4-10%). Either qualitative or quantitative differences, mainly related to the lutein proportion, were found among three batches of jackfruit. Since the fruits from batch A showed significantly lower contents for almost all carotenoids, it also had the lowest total carotenoid content (34.1 microg/100 g) and provitamin A value, whereas the total carotenoid ranged from 129.0 to 150.3 microg/100 g in the other batches. The provitamin A values from batches B and C were 3.3 and 4.3 microg RAE/100 g, respectively. The carotenoid composition of jackfruit was successfully determined, where 14 of the 18 identified carotenoids were reported for first time. Differences among batches may be due to genetic and/or agricultural factors.

Chemically bonded stationary phases that use synthetic hosts containing aromatic binding clefts: HPLC analysis of nitro-substituted polycyclic aromatic hydrocarbons.

PubMed Central

Zimmerman, S C; Saionz, K W; Zeng, Z

1993-01-01

The synthesis of hosts with improved binding affinities for nitroaromatic guests is described. Association constants for several host-guest complexes were measured in chloroform solution and ranged over three orders of magnitude. Two hosts were covalently linked to silica gel to produce chemically bonded stationary phases for HPLC. The use of these phases for HPLC analysis of nitro-substituted polycyclic aromatic hydrocarbons is discussed. PMID:8433981

Pigmented Cells in the Pineal Gland of Female Viscacha (Lagostomus maximus maximus): A Histochemical and Ultrastructural Study

PubMed Central

Busolini, Fabricio Ivan; Rodríguez, Graciela Beatriz; Filippa, Verónica Palmira

2017-01-01

The presence of pigment has been demonstrated in different nervous structures such as those of retina, substantia nigra, and locus coeruleus. These pigments have also been described in the pineal gland of different mammal species. Histochemical and ultrastructural studies of the pineal gland of female viscacha (Lagostomus maximus maximus) were performed to analyze the presence of pigmented cells under natural conditions and to evaluate a probable relation between pigment content and glandular activity during pregnancy. The following techniques were applied: hematoxylin-eosin, phosphotungstic acid-hematoxylin, Masson-Fontana silver, DOPA histochemistry, Schmorl's reaction and toluidine blue. Estradiol and progesterone serum levels were determined by RIA. The ultrastructural features of the pineal pigment granules were also analyzed. Pigment granules were observed in a random distribution, but the pigmented cells were frequently found near blood vessels. The pineal pigment was histochemically identified as melanin. Differences in the amount of pigmented cells were found between pregnant and nonpregnant viscachas. The ultrastructural analysis revealed the presence of premelanosomes and melanosomes. Estradiol and progesterone levels vary during pregnancy. In conclusion, the changes in the amount of pigment content and hormone levels may indicate that the pineal gland of female viscacha is susceptible to endocrine variations during pregnancy. PMID:29391866

[Analysis of different forms Linderae Radix based on HPLC and NIRS fingerprints].

PubMed

Du, Wei-Feng; Yue, Xian-Ke; Wu, Yao; Ge, Wei-Hong; Lu, Tu-Lin; Wang, Zhi-Min

2016-10-01

Three different forms of Linderae Radix were evaluated by HPLC combined with NIRS fingerprint. The Linderae Radix was divided into three forms, including spindle root, straight root and old root. The HPLC fingerprints were developed, and then cluster analysis was performed using the SPSS software. The near-infrared spectra of Linderae Radix was collected, and then established the discriminant analysis model. The similarity values of the spindle root and straight root all were above 0.990, while the similarity value of the old root was less than 0.850. Two forms of Linderae Radix were obviously divided into three parts by the NIRS model and Cluster analysis. The results of HPLC and FT-NIR analysis showed the quality of Linderae Radix old root was different from the spindle root and straight root. The combined use of the two methods could identify different forms of Linderae Radix quickly and accurately. Copyright© by the Chinese Pharmaceutical Association.

Down-regulated PAR-2 is associated in part with interrupted melanosome transfer in pigmented basal cell epithelioma.

PubMed

Sakuraba, Kazuko; Hayashi, Nobukazu; Kawashima, Makoto; Imokawa, Genji

2004-08-01

In pigmented basal cell epithelioma (BCE), there seems to be an abnormal transfer of melanized melanosomes from proliferating melanocytes to basaloid tumor cells. In this study, the interruption of that melanosome transfer was studied with special respect to the altered function of a phagocytic receptor, protease-activated receptor (PAR)-2 in the basaloid tumor cells. We used electron microscopy to clarify the disrupted transfer at the ultrastructural level and then performed immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) to examine the regulation of a phagocytic receptor, PAR-2, expressed on basaloid tumor cells. Electron microscopic analysis revealed that basaloid tumor cells of pigmented BCE have a significantly lower population of melanosomes ( approximately 16.4%) than do normal keratinocytes located in the perilesional normal epidermis ( approximately 91.0%). In contrast, in pigmented seborrheic keratosis (SK), a similarly pigmented epidermal tumor, the distribution of melanin granules does not differ between the lesional ( approximately 93.9%) and the perilesional normal epidermis ( approximately 92.2 %), indicating that interrupted melanosome transfer occurs in BCE but not in all pigmented epithelial tumors. RT-PCR analysis demonstrated that the expression of PAR-2 mRNA transcripts in basaloid cells is significantly decreased in pigmented BCE compared with the perilesional normal epidermis. In contrast, in pigmented SK, where melanosome transfer to basaloid tumor cells is not interrupted, the expression of PAR-2 mRNA transcripts is comparable between the basaloid tumor cells and the perilesional normal epidermis. Immunohistochemistry demonstrated that basaloid cells in pigmented BCE have less immunostaining for PAR-2 than do keratinocytes in the perilesional normal epidermis whereas in pigmented SK, there is no difference in immunostaining for PAR-2 between the basaloid tumor and the perilesional normal epidermis. These findings suggest that the decreased expression of PAR-2 in the basaloid cells is associated in part with the observed interruption of melanosome transfer in pigmented BCE.

Selenium speciation analysis of Misgurnus anguillicaudatus selenoprotein by HPLC-ICP-MS and HPLC-ESI-MS/MS

USDA-ARS?s Scientific Manuscript database

Analytical methods for selenium (Se) speciation were developed using high performance liquid chromatography (HPLC) coupled to either inductively coupled plasma mass spectrometry (ICP-MS) or electrospray ionization tandem mass spectrometry (ESI-MS/MS). Separations of selenomethionine (Se-Met) and sel...

Determination of vitamin E and carotenoid pigments by high performance liquid chromatography in shell of Chionoecetes opilio.

PubMed

Vilasoa-Martínez, María; Calaza-Ramos, Carina; López-Hernández, Julia; Lage-Yusty, María Asunción; Losada, Perfecto Paseiro; Rodríguez-Bernaldo de Quirós, Ana

2008-06-09

This study reports the optimization of a method for the determination of vitamin E and carotenoids in shells of Chionoecetes opilio samples by online HPLC coupled with UV-vis and fluorescence detectors. The carotenoids were determined with diode-array detector (lambda 450 nm) and vitamin E with fluorescence detection (lambda(ex) 288, lambda(em) 331 nm). Two extractions methods were compared, saponification followed by an extraction step and a simple extraction with acetone. The last one was selected because allows to determine all compounds. Linearity, precisions and recoveries achieved for all compounds were satisfactory. Mean concentrations (mg per 100 g dry weight) were; 23.3 for vitamin E, 9.49 for astaxanthin and 0.2 mg for beta-carotene.

Evaluation of multiple reaction monitoring cubed for the analysis of tachykinin related peptides in rat spinal cord using a hybrid triple quadrupole-linear ion trap mass spectrometer.

PubMed

Pailleux, Floriane; Beaudry, Francis

2014-02-01

Targeted peptide methods generally use HPLC-MS/MRM approaches. Although dependent on the instrumental resolution, interferences may occur while performing analysis of complex biological matrices. HPLC-MS/MRM(3) is a technique, which provides a significantly better selectivity, compared with HPLC-MS/MRM assay. HPLC-MS/MRM(3) allows the detection and quantitation by enriching standard MRM with secondary product ions that are generated within the linear ion trap. Substance P (SP) and neurokinin A (NKA) are tachykinin peptides playing a central role in pain transmission. The objective of this study was to verify whether HPLC-MS/MRM(3) could provide significant advantages over a more traditional HPLC-MS/MRM assay for the quantification of SP and NKA in rat spinal cord. The results suggest that reconstructed MRM(3) chromatograms display significant improvements with the nearly complete elimination of interfering peaks but the sensitivity (i.e. signal-to-noise ratio) was severely reduced. The precision (%CV) observed was between 3.5% and 24.1% using HPLC-MS/MRM and in the range of 4.3-13.1% with HPLC-MS/MRM(3), for SP and NKA. The observed accuracy was within 10% of the theoretical concentrations tested. HPLC-MS/MRM(3) may improve the assay sensitivity to detect difference between samples by reducing significantly the potential of interferences and therefore reduce instrumental errors. Copyright © 2014 Elsevier B.V. All rights reserved.

Melanogenesis of murine melanoma cells induced by hesperetin, a Citrus hydrolysate-derived flavonoid.

PubMed

Huang, Yu-Chun; Liu, Kao-Chih; Chiou, Yi-Ling

2012-03-01

Melanogenesis is a complex process that modulates skin pigmentation to defend photodamage. Citrus is the most widely produced fruit crop in the world. People ingest various citrus fruits in their common diets. In the present study, the acid-hydrolyzed and un-hydrolyzed extracts of orange-type citrus fruits were subjected to analyze flavonoid compositions and assess their effects on melanin synthesis in murine B16-F10 melanoma cells. The acid-hydrolyzed extracts of Citrus sinensis, C. reticulata, and C. aurantium enhanced melanin production. Based on high-performance liquid chromatography (HPLC) analysis, the most abundant flavonoids that were found in citrus hydrolyzed extracts were hesperetin and naringenin. Hesperetin exhibited the most potent activity on melanin synthesis and induced tyrosinase and microphthalmia-associated transcription factor (MITF) expression. Moreover, hesperetin stimulated the activation of mitogen-activated protein kinases (MAPKs), phosphorylation of cAMP-responsive element binding protein (CREB) and glycogen synthase kinase-3β (GSK3β), and subsequently induced the accumulation of β-catenin. This study suggests that the citrus constituent hesperetin might have protective melanogenic potential as a cosmeceutical agent against skin photodamage. Copyright © 2012 Elsevier Ltd. All rights reserved.

Phytoplankton variability and community structure in relation to hydrographic features in the NE Aegean frontal area (NE Mediterranean Sea)

NASA Astrophysics Data System (ADS)

Lagaria, A.; Mandalakis, M.; Mara, P.; Frangoulis, C.; Karatsolis, B.-Th.; Pitta, P.; Triantaphyllou, M.; Tsiola, A.; Psarra, S.

2017-10-01

The structure of phytoplankton community in the salinity-stratified Northeastern Aegean frontal area adjacent to the Dardanelles Straits was investigated on a seasonal basis (autumn, spring and summer) and in relation to circulating water masses: the modified Black Sea Water (BSW) and the Levantine Water (LW). By employing High Performance Liquid Chromatography (HPLC) for the analysis of phytoplankton pigments in conjunction with conventional cell counting methodologies (i.e. inverted light microscopy, flow cytometry) and primary production measurements, a comprehensive qualitative and quantitative characterization of phytoplankton community composition and its activity was conducted. Chlorophyll-a normalized production and estimated growth rates presented the highest values within the 'fresh' BSW mass during summer, though generally growth rates were low (<0.4 d-1) at all seasons. The spatiotemporal variation of BSW outflow was found to greatly affect the relative contribution of pico-, nano- and micro-phytoplankton to total phytoplankton biomass and production. Large cell organisms, and in particular diatoms, were closely associated with the surface BSW masses outflowing from the Straits. Our results showed that all phytoplankton size components were significant over time and space suggesting a rather multivorous food web functioning of the system.

Micro-Raman spectroscopy and chemometrical analysis for the distinction of copper phthalocyanine polymorphs in paint layers.

PubMed

Defeyt, C; Van Pevenage, J; Moens, L; Strivay, D; Vandenabeele, P

2013-11-01

In art analysis, copper phthalocyanine (CuPc) is often identified as an important pigment (PB15) in 20th century artworks. Raman spectroscopy is a very valuable technique for the detection of this pigment in paint systems. However, PB15 is used in different polymorphic forms and identification of the polymorph could retrieve information on the production process of the pigment at the moment. Raman spectroscopy, being a molecular spectroscopic method of analysis, is able to discriminate between polymorphs of crystals. However, in the case of PB15, spectral interpretation is not straightforward, and Raman data treatment requires some improvements concerning the PB15 polymorphic discrimination in paints. Here, Raman spectroscopy is combined with chemometrical analysis in order to develop a procedure allowing us to identify the PB15 crystalline structure in painted layers and in artworks. The results obtained by Linear Discriminant Analysis (LDA), using intensity ratios as variables, demonstrate the ability of this procedure to predict the crystalline structure of a PB15 pigment in unknown paint samples. Copyright © 2013 Elsevier B.V. All rights reserved.

Trace analysis of pollutants in water using the photothermal interferometry as HPLC detector.

PubMed

Seidel, B S; Dübel, O; Faubel, W; Ache, H J

1996-03-01

A new procedure including high performance liquid chromatography in combination with photothermal interference spectroscopy as detection device (HPLC/PIS) has been proposed, optimized and its figures of merit for pesticide residue analysis are shown. The flowing sample under study is set in one arm of a Mach-Zehnder interferometer, and its refractive index is modulated by a periodically chopped continuous wave argon ion laser. As chopper, an acousto optical modulator has been introduced to switch the excitation laser beam between different lines (457 nm, 488 nm, 514 nm) simultaneously. Thus a multi component analysis can be realized either by using an HPLC-system in front of the PIS device or by a multi line Ar(+)-laser, directly. The limit of detection of the HPLC/PIS system reached 71 microg/l of the pesticide di-nitro-ortho-cresol (DNOC).

Two-dimensional fingerprinting approach for comparison of complex substances analysed by HPLC-UV and fluorescence detection.

PubMed

Ni, Yongnian; Liu, Ying; Kokot, Serge

2011-02-07

This work is concerned with the research and development of methodology for analysis of complex mixtures such as pharmaceutical or food samples, which contain many analytes. Variously treated samples (swill washed, fried and scorched) of the Rhizoma atractylodis macrocephalae (RAM) traditional Chinese medicine (TCM) as well as the common substitute, Rhizoma atractylodis (RA) TCM were chosen as examples for analysis. A combined data matrix of chromatographic 2-D HPLC-DAD-FLD (two-dimensional high performance liquid chromatography with diode array and fluorescence detectors) fingerprint profiles was constructed with the use of the HPLC-DAD and HPLC-FLD individual data matrices; the purpose was to collect maximum information and to interpret this complex data with the use of various chemometrics methods e.g. the rank-ordering multi-criteria decision making (MCDM) PROMETHEE and GAIA, K-nearest neighbours (KNN), partial least squares (PLS), back propagation-artificial neural networks (BP-ANN) methods. The chemometrics analysis demonstrated that the combined 2-D HPLC-DAD-FLD data matrix does indeed provide more information and facilitates better performing classification/prediction models for the analysis of such complex samples as the RAM and RA ones noted above. It is suggested that this fingerprint approach is suitable for analysis of other complex, multi-analyte substances.

Novel Pathways of Nitroaromatic Metabolism: Hydroxylamine Formation, Reactivity and Potential for Ring Fission for Destruction of TNT-CU1214

DTIC Science & Technology

2005-08-15

phase was terminated when HPLC analysis showed that the initial TNT was in the form of DHA6NT and aminophenols , products that have been reported...terminated when HPLC analysis showed that the initial TNT was in the form of DHA6NT and aminophenols , products that have been reported previously (2...Department of Defense HPLC High Performance Liquid Chromatography IPTG Isopropyl-β-D-1-thiogalactopyranoside LB Luria-Bertani Broth NB Nitrobenzene

Pigment granule translocation in red ovarian chromatophores from the palaemonid shrimp Macrobrachium olfersi (Weigmann, 1836): functional roles for the cytoskeleton and its molecular motors.

PubMed

Milograna, Sarah Ribeiro; Ribeiro, Márcia Regina; Baqui, Munira Muhammad Abdel; McNamara, John Campbell

2014-12-01

The binding of red pigment concentrating hormone (RPCH) to membrane receptors in crustacean chromatophores triggers Ca²⁺/cGMP signaling cascades that activate cytoskeletal motors, driving pigment granule translocation. We investigate the distributions of microfilaments and microtubules and their associated molecular motors, myosin and dynein, by confocal and transmission electron microscopy, evaluating a functional role for the cytoskeleton in pigment translocation using inhibitors of polymer turnover and motor activity in vitro. Microtubules occupy the chromatophore cell extensions whether the pigment granules are aggregated or dispersed. The inhibition of microtubule turnover by taxol induces pigment aggregation and inhibits re-dispersion. Phalloidin-FITC actin labeling, together with tannic acid fixation and ultrastructural analysis, reveals that microfilaments form networks associated with the pigment granules. Actin polymerization induced by jasplaquinolide strongly inhibits RPCH-induced aggregation, causes spontaneous pigment dispersion, and inhibits pigment re-dispersion. Inhibition of actin polymerization by latrunculin-A completely impedes pigment aggregation and re-dispersion. Confocal immunocytochemistry shows that non-muscle myosin II (NMMII) co-localizes mainly with pigment granules while blebbistatin inhibition of NMMII strongly reduces the RPCH response, also inducing spontaneous pigment dispersion. Myosin II and dynein also co-localize with the pigment granules. Inhibition of dynein ATPase by erythro-9-(2-hydroxy-3-nonyl) adenine induces aggregation, inhibits RPCH-triggered aggregation, and inhibits re-dispersion. Granule aggregation and dispersion depend mainly on microfilament integrity although microtubules may be involved. Both cytoskeletal polymers are functional only when subunit turnover is active. Myosin and dynein may be the molecular motors that drive pigment aggregation. These mechanisms of granule translocation in crustacean chromatophores share various features with those of vertebrate pigment cells. Copyright © 2014 Elsevier Inc. All rights reserved.

Fingerprint chromatogram analysis of Pseudostellaria heterophylla (Miq.) Pax root by high performance liquid chromatography.

PubMed

Han, Chao; Chen, Junhui; Chen, Bo; Lee, Frank Sen-Chun; Wang, Xiaoru

2006-09-01

A simple and reliable high performance liquid chromatographic (HPLC) method has been developed and validated for the fingerprinting of extracts from the root of Pseudostellaria heterophylla (Miq.) Pax. HPLC with gradient elution was performed on an authentic reference standard of powdered P. heterophylla (Miq.) Pax root and 11 plant samples of the root were collected from different geographic locations. The HPLC chromatograms have been standardized through the selection and identification of reference peaks and the normalization of retention times and peak intensities of all the common peaks. The standardized HPLC fingerprints show high stability and reproducibility, and thus can be used effectively for the screening analysis or quality assessment of the root or its derived products. Similarity index calculations based on cosine angle values or correlation methods have been performed on the HPLC fingerprints. As a group, the fingerprints of the P. heterophylla (Miq.) Pax samples studied are highly correlated with closely similar fingerprints. Within the group, the samples can be further divided into subgroups based on hierarchical clustering analysis (HCA). Sample grouping based on HCA coincides nicely with those based on the geographical origins of the samples. The HPLC fingerprinting techniques thus have high potential in authentication or source-tracing types of applications.

Comparison of transferrin isoform analysis by capillary electrophoresis and HPLC for screening congenital disorders of glycosylation.

PubMed

Dave, Mihika B; Dherai, Alpa J; Udani, Vrajesh P; Hegde, Anaita U; Desai, Neelu A; Ashavaid, Tester F

2018-01-01

Transferrin, a major glycoprotein has different isoforms depending on the number of sialic acid residues present on its oligosaccharide chain. Genetic variants of transferrin as well as the primary (CDG) & secondary glycosylation defects lead to an altered transferrin pattern. Isoform analysis methods are based on charge/mass variations. We aimed to compare the performance of commercially available capillary electrophoresis CDT kit for diagnosing congenital disorders of glycosylation with our in-house optimized HPLC method for transferrin isoform analysis. The isoform pattern of 30 healthy controls & 50 CDG-suspected patients was determined by CE using a Carbohydrate-Deficient Transferrin kit. The results were compared with in-house HPLC-based assay for transferrin isoforms. Transferrin isoform pattern for healthy individuals showed a predominant tetrasialo transferrin fraction followed by pentasialo, trisialo, and disialotransferrin. Two of 50 CDG-suspected patients showed the presence of asialylated isoforms. The results were comparable with isoform pattern obtained by HPLC. The commercial controls showed a <20% CV for each isoform. Bland Altman plot showed the difference plot to be within +1.96 with no systemic bias in the test results by HPLC & CE. The CE method is rapid, reproducible and comparable with HPLC and can be used for screening Glycosylation defects. © 2017 Wiley Periodicals, Inc.

Detection of pigment network in dermoscopy images using supervised machine learning and structural analysis.

PubMed

García Arroyo, Jose Luis; García Zapirain, Begoña

2014-01-01

By means of this study, a detection algorithm for the "pigment network" in dermoscopic images is presented, one of the most relevant indicators in the diagnosis of melanoma. The design of the algorithm consists of two blocks. In the first one, a machine learning process is carried out, allowing the generation of a set of rules which, when applied over the image, permit the construction of a mask with the pixels candidates to be part of the pigment network. In the second block, an analysis of the structures over this mask is carried out, searching for those corresponding to the pigment network and making the diagnosis, whether it has pigment network or not, and also generating the mask corresponding to this pattern, if any. The method was tested against a database of 220 images, obtaining 86% sensitivity and 81.67% specificity, which proves the reliability of the algorithm. © 2013 The Authors. Published by Elsevier Ltd. All rights reserved.

Methods of analysis by the U.S. Geological Survey Organic Geochemistry Research Group; determination of chloroacetanilide herbicide metabolites in water using high-performance liquid chromatography-diode array detection and high-performance liquid chromatography/mass spectrometry

USGS Publications Warehouse

Zimmerman, L.R.; Hostetler, K.A.; Thurman, E.M.

2000-01-01

Analytical methods using high-performance liquid chromatography-diode array detection (HPLC-DAD) and high-performance liquid chromatography/mass spectrometry (HPLC/MS) were developed for the analysis of the following chloroacetanilide herbicide metabolites in water: acetochlor ethanesulfonic acid (ESA), acetochlor oxanilic acid (OXA), alachlor ESA, alachlor OXA, metolachlor ESA, and metolachlor OXA. Good precision and accuracy were demonstrated for both the HPLC-DAD and HPLC/MS methods in reagent water, surface water, and ground water. The mean HPLC-DAD recoveries of the chloroacetanilide herbicide metabolites from water samples spiked at 0.25, 0.50, and 2.0 mg/L (micrograms per liter) ranged from 84 to 112 percent, with relative standard deviations of 18 percent or less. The mean HPLC/MS recoveries of the metabolites from water samples spiked at 0.05, 0.20, and 2.0 mg/L ranged from 81 to 125 percent, with relative standard deviations of 20 percent or less. The limit of quantitation (LOQ) for all metabolites using the HPLC-DAD method was 0.20 mg/L, whereas the LOQ using the HPLC/MS method was 0.05 mg/L. These metabolite-determination methods are valuable for acquiring information about water quality and the fate and transport of the parent chloroacetanilide herbicides in water.

Comparison of the phenolic composition of fruit juices by single step gradient HPLC analysis of multiple components versus multiple chromatographic runs optimised for individual families.

PubMed

Bremner, P D; Blacklock, C J; Paganga, G; Mullen, W; Rice-Evans, C A; Crozier, A

2000-06-01

After minimal sample preparation, two different HPLC methodologies, one based on a single gradient reversed-phase HPLC step, the other on multiple HPLC runs each optimised for specific components, were used to investigate the composition of flavonoids and phenolic acids in apple and tomato juices. The principal components in apple juice were identified as chlorogenic acid, phloridzin, caffeic acid and p-coumaric acid. Tomato juice was found to contain chlorogenic acid, caffeic acid, p-coumaric acid, naringenin and rutin. The quantitative estimates of the levels of these compounds, obtained with the two HPLC procedures, were very similar, demonstrating that either method can be used to analyse accurately the phenolic components of apple and tomato juices. Chlorogenic acid in tomato juice was the only component not fully resolved in the single run study and the multiple run analysis prior to enzyme treatment. The single run system of analysis is recommended for the initial investigation of plant phenolics and the multiple run approach for analyses where chromatographic resolution requires improvement.

Root Cause Analysis of Quality Defects Using HPLC-MS Fingerprint Knowledgebase for Batch-to-batch Quality Control of Herbal Drugs.

PubMed

Yan, Binjun; Fang, Zhonghua; Shen, Lijuan; Qu, Haibin

2015-01-01

The batch-to-batch quality consistency of herbal drugs has always been an important issue. To propose a methodology for batch-to-batch quality control based on HPLC-MS fingerprints and process knowledgebase. The extraction process of Compound E-jiao Oral Liquid was taken as a case study. After establishing the HPLC-MS fingerprint analysis method, the fingerprints of the extract solutions produced under normal and abnormal operation conditions were obtained. Multivariate statistical models were built for fault detection and a discriminant analysis model was built using the probabilistic discriminant partial-least-squares method for fault diagnosis. Based on multivariate statistical analysis, process knowledge was acquired and the cause-effect relationship between process deviations and quality defects was revealed. The quality defects were detected successfully by multivariate statistical control charts and the type of process deviations were diagnosed correctly by discriminant analysis. This work has demonstrated the benefits of combining HPLC-MS fingerprints, process knowledge and multivariate analysis for the quality control of herbal drugs. Copyright © 2015 John Wiley & Sons, Ltd.

Tentative characterization of precursor compounds and co-factors of pigment formation in production of 'wu mi' from Vaccinium bracteatum Thunb. Leaves.

PubMed

Fan, Mingcong; Fan, Yihui; Huang, Weiping; Wang, Li; Li, Yan; Qian, Haifeng; Zhang, Hui; Qi, Xiguang

2018-10-01

Vaccinium bracteatum leaves (VBTL) are traditionally used in China to dye rice grains, which assume a deep blue color, named 'Wu mi'. Information on the mechanism of pigment formation is limited. In this study, CIELAB color space parameters were used to represent the color of 'Wu mi'. Precursor compounds of pigments formed during the dyeing process were identified by UPLC Q-TOF MS analysis. The changes in co-factors for pigment formation in VBTL were measured at different growth stages. The L ∗ and b ∗ values of dyed rice increased as the leaves aged, whereas a ∗ values showed irregular changes. Six compounds were tentatively identified as pigment precursors by UPLC Q-TOF MS analysis. The pH and β-glucosidase activity at different growth stages of VBTL were indicated to be crucial co-factors for pigment formation. A tentative hypothesis is presented that iridoid glycosides are hydrolyzed by acids and β-glucosidases to form a dialdehyde structure that binds covalently with amino residues of lysine side chains in rice protein molecules. Copyright © 2018 Elsevier Ltd. All rights reserved.

Red fluorescent protein responsible for pigmentation in trematode-infected Porites compressa tissues.

PubMed

Palmer, Caroline V; Roth, Melissa S; Gates, Ruth D

2009-02-01

Reports of coral disease have increased dramatically over the last decade; however, the biological mechanisms that corals utilize to limit infection and resist disease remain poorly understood. Compromised coral tissues often display non-normal pigmentation that potentially represents an inflammation-like response, although these pigments remain uncharacterized. Using spectral emission analysis and cryo-histological and electrophoretic techniques, we investigated the pink pigmentation associated with trematodiasis, infection with Podocotyloides stenometre larval trematode, in Porites compressa. Spectral emission analysis reveals that macroscopic areas of pink pigmentation fluoresce under blue light excitation (450 nm) and produce a broad emission peak at 590 nm (+/-6) with a 60-nm full width at half maximum. Electrophoretic protein separation of pigmented tissue extract confirms the red fluorescence to be a protein rather than a low-molecular-weight compound. Histological sections demonstrate green fluorescence in healthy coral tissue and red fluorescence in the trematodiasis-compromised tissue. The red fluorescent protein (FP) is limited to the epidermis, is not associated with cells or granules, and appears unstructured. These data collectively suggest that the red FP is produced and localized in tissue infected by larval trematodes and plays a role in the immune response in corals.

[HPLC specific chromatogram of Lamiophlomis Herba and its counterfeit and determination of four effective components].

PubMed

Zan, Ke; Jiao, Xing-Ping; Guo, Li-Nong; Zheng, Jian; Ma, Shuang-Cheng

2016-06-01

This study is to establish the HPLC specific chromatogram and determine four main effective components of Lamiophlomis Herba and its counterfeit.Chlorogenic acid, forsythoside B, acteoside and luteoloside were reference substance.HPLC analysis was performed on a Waters XSelect C₁₈ column (4.6 mm×250 mm,5 μm).The mobile phase was acetonitrile-0.5% phosphoric acid solution (18∶82) with isocratic elution.The flow rate was 1.0 mL•min⁻¹, the detection wavelength was 332 nm and the column temperature was 30 ℃.Chemometrics software Chempattern was employed to analyze the research data.HPLC specific chromatogram of Lamiophlomis Herba from different samples were of high similarity, but the similarity of the HPLC specific chromatogram of its counterfeit were less than 0.65.Both of cluster and principal component analysis can distinguish certified products and adulterants.The HPLC specific chromatogram and contents of four effective components can be used for the quality control of Lamiophlomis Herba and its preparations.It provided scientific basis to standardize the use of the crude drug. Copyright© by the Chinese Pharmaceutical Association.

Major and Modified Nucleosides, RNA, and DNA

NASA Astrophysics Data System (ADS)

Gehrke, Charles W.; Kuo, Kenneth C.

Most analytical chemists are well aware of the rapid rate of development of high-performance liquid chromatography (HPLC) over the past 5 years. A number of articles have been published in Analytical Chemistry on different topics in HPLC and many papers appear in the chromatographic journals. Some books also have been published covering this subject. HPLC has proved to be a very effective, broadly applicable chromatographic method for the separation and analysis of complex molecules in fields as diverse as biochemistry and environmental, pharmaceutical, medical, and polymer chemistry. HPLC is now having a major impact on the clinical and research aspects of medical biochemistry. Although the contributions of HPLC to other disciplines generally complements gas-liquid chromatography, this method is destined to play a much greater role in medical and biochemical research. This is because many of the biomolecules, owing to their molecular complexity and size, are thermally unstable or nonvolatile, preventing or complicating an analysis by GC. A major factor contributing to the powerful advances in biomedical liquid chromatography is the development of reversed-phase high-performance liquid chromatography (RP-HPLC) using n-alkyl and phenyl chemically bonded substrates.

Raman analysis of cobalt blue pigment in blue and white porcelain: A reassessment.

PubMed

Jiang, Xiaochenyang; Ma, Yanying; Chen, Yue; Li, Yuanqiu; Ma, Qinglin; Zhang, Zhaoxia; Wang, Changsui; Yang, Yimin

2018-02-05

Cobalt blue is a famous pigment in human history. In the past decade it is widely reported that the cobalt aluminate has been detected in ancient ceramics as blue colorant in glaze, yet the acquired Raman spectra are incredibly different from that of synthesised references, necessitating a reassessment of such contradictory scenario with more accurate analytic strategies. In this study, micro-Raman spectroscopy (MRS) and scanning electron microscopy (SEM) in association with energy dispersive spectrometry (EDS) were performed on under-glaze cobalt pigments from one submerged blue and white porcelain shard dated from Wanli reign (1573-1620CE) of Ming dynasty (1365-1644CE) excavated at Nan'ao I shipwreck off the southern coast of China. The micro-structural inspection reveals that the pigment particles have characteristics of small account, tiny size, heterogeneously distribution, and more importantly, been completely enwrapped by well-developed anorthite crystals in the glaze, indicating that the signals recorded in previous publications are probably not from cobalt pigments themselves but from outside thickset anorthite shell. The further spectromicroscopic analyses confirm this presumption when the accurate spectra of cobalt aluminate pigment and surrounding anorthite were obtained separately with precise optical positioning. Accordingly, we reassess and clarify the previous Raman studies dedicated to cobalt blue pigment in ancient ceramics, e.g. cobalt blue in celadon glaze, and in turn demonstrate the superiority and necessity of coupling spectroscopic analysis with corresponding structure observation, especially in the characterization of pigments from complicated physico-chemical environment like antiquities. Thus, this study promotes a better understanding of Raman spectroscopy study of cobalt blue pigments in art and archaeology field. Copyright © 2017 Elsevier B.V. All rights reserved.

Raman analysis of cobalt blue pigment in blue and white porcelain: A reassessment

NASA Astrophysics Data System (ADS)

Jiang, Xiaochenyang; Ma, Yanying; Chen, Yue; Li, Yuanqiu; Ma, Qinglin; Zhang, Zhaoxia; Wang, Changsui; Yang, Yimin

2018-02-01

Cobalt blue is a famous pigment in human history. In the past decade it is widely reported that the cobalt aluminate has been detected in ancient ceramics as blue colorant in glaze, yet the acquired Raman spectra are incredibly different from that of synthesised references, necessitating a reassessment of such contradictory scenario with more accurate analytic strategies. In this study, micro-Raman spectroscopy (MRS) and scanning electron microscopy (SEM) in association with energy dispersive spectrometry (EDS) were performed on under-glaze cobalt pigments from one submerged blue and white porcelain shard dated from Wanli reign (1573-1620 CE) of Ming dynasty (1365-1644 CE) excavated at Nan'ao I shipwreck off the southern coast of China. The micro-structural inspection reveals that the pigment particles have characteristics of small account, tiny size, heterogeneously distribution, and more importantly, been completely enwrapped by well-developed anorthite crystals in the glaze, indicating that the signals recorded in previous publications are probably not from cobalt pigments themselves but from outside thickset anorthite shell. The further spectromicroscopic analyses confirm this presumption when the accurate spectra of cobalt aluminate pigment and surrounding anorthite were obtained separately with precise optical positioning. Accordingly, we reassess and clarify the previous Raman studies dedicated to cobalt blue pigment in ancient ceramics, e.g. cobalt blue in celadon glaze, and in turn demonstrate the superiority and necessity of coupling spectroscopic analysis with corresponding structure observation, especially in the characterization of pigments from complicated physico-chemical environment like antiquities. Thus, this study promotes a better understanding of Raman spectroscopy study of cobalt blue pigments in art and archaeology field.

High-speed counter-current chromatography coupled online to high performance liquid chromatography-diode array detector-mass spectrometry for purification, analysis and identification of target compounds from natural products.

PubMed

Liang, Xuejuan; Zhang, Yuping; Chen, Wei; Cai, Ping; Zhang, Shuihan; Chen, Xiaoqin; Shi, Shuyun

2015-03-13

A challenge in coupling high-speed counter-current chromatography (HSCCC) online with high performance liquid chromatography (HPLC) for purity analysis was their time incompatibility. Consequently, HSCCC-HPLC was conducted by either controlling HPLC analysis time and HSCCC flow rate or using stop-and-go scheme. For natural products containing compounds with a wide range of polarities, the former would optimize experimental conditions, while the latter required more time. Here, a novel HSCCC-HPLC-diode array detector-mass spectrometry (HSCCC-HPLC-DAD-MS) was developed for undisrupted purification, analysis and identification of multi-compounds from natural products. Two six-port injection valves and a six-port switching valve were used as interface for collecting key HSCCC effluents alternatively for HPLC-DAD-MS analysis and identification. The ethyl acetate extract of Malus doumeri was performed on the hyphenated system to verify its efficacy. Five main flavonoids, 3-hydroxyphloridzin (1), phloridzin (2), 4',6'-dihydroxyhydrochalcone-2'-O-β-D-glucopyranoside (3, first found in M. doumeri), phloretin (4), and chrysin (5), were purified with purities over 99% by extrusion elution and/or stepwise elution mode in two-step HSCCC, and 25mM ammonium acetate solution was selected instead of water to depress emulsification in the first HSCCC. The online system shortened manipulation time largely compared with off-line analysis procedure and stop-and-go scheme. The results indicated that the present method could serve as a simple, rapid and effective way to achieve target compounds with high purity from natural products. Copyright © 2015 Elsevier B.V. All rights reserved.

Gingival Pigmentation Affected by Smoking among Different Age Groups: A Quantitative Analysis of Gingival Pigmentation Using Clinical Oral Photographs.

PubMed

Kato, Tomotaka; Mizutani, Shinsuke; Takiuchi, Hiroya; Sugiyama, Seiichi; Hanioka, Takashi; Naito, Toru

2017-08-04

The presence of any age-related differences in gingival pigmentation associated with smoking, particularly in a young population, remains to be fully investigated. The purpose of this study was to determine the age-related differences in smoking gingival pigmentation. Gingival pigmentation was analyzed using the gingival melanosis record (GMR) and Hedin's classification with frontal oral photographs taken at 16 dental offices in Japan. Participants were categorized into 10-year age groups, and their baseline photographs were compared. In addition, to evaluate the effect of smoking cessation on gingival pigmentation, subjects were divided into a former smoker group (stopped smoking) and current smoker group. A total of 259 patients 19 to 79 years of age were analyzed. People in their 30s showed the most widespread gingival pigmentation. In addition, subjects in their 20s showed a weak effect of smoking cessation on gingival pigmentation. These findings suggested that the gingival pigmentation induced by smoking was more remarkable in young people than in middle-aged people. This information may be useful for anti-smoking education, especially among young populations with a high affinity for smoking.

The genes and enzymes of the carotenoid metabolic pathway in Vitis vinifera L.

PubMed Central

2012-01-01

Background Carotenoids are a heterogeneous group of plant isoprenoids primarily involved in photosynthesis. In plants the cleavage of carotenoids leads to the formation of the phytohormones abscisic acid and strigolactone, and C13-norisoprenoids involved in the characteristic flavour and aroma compounds in flowers and fruits and are of specific importance in the varietal character of grapes and wine. This work extends the previous reports of carotenoid gene expression and photosynthetic pigment analysis by providing an up-to-date pathway analysis and an important framework for the analysis of carotenoid metabolic pathways in grapevine. Results Comparative genomics was used to identify 42 genes putatively involved in carotenoid biosynthesis/catabolism in grapevine. The genes are distributed on 16 of the 19 chromosomes and have been localised to the physical map of the heterozygous ENTAV115 grapevine sequence. Nine of the genes occur as single copies whereas the rest of the carotenoid metabolic genes have more than one paralogue. The cDNA copies of eleven corresponding genes from Vitis vinifera L. cv. Pinotage were characterised, and four where shown to be functional. Microarrays provided expression profiles of 39 accessions in the metabolic pathway during three berry developmental stages in Sauvignon blanc, whereas an optimised HPLC analysis provided the concentrations of individual carotenoids. This provides evidence of the functioning of the lutein epoxide cycle and the respective genes in grapevine. Similarly, orthologues of genes leading to the formation of strigolactone involved in shoot branching inhibition were identified: CCD7, CCD8 and MAX1. Moreover, the isoforms typically have different expression patterns, confirming the complex regulation of the pathway. Of particular interest is the expression pattern of the three VvNCEDs: Our results support previous findings that VvNCED3 is likely the isoform linked to ABA content in berries. Conclusions The carotenoid metabolic pathway is well characterised, and the genes and enzymes have been studied in a number of plants. The study of the 42 carotenoid pathway genes of grapevine showed that they share a high degree of similarity with other eudicots. Expression and pigment profiling of developing berries provided insights into the most complete grapevine carotenoid pathway representation. This study represents an important reference study for further characterisation of carotenoid biosynthesis and catabolism in grapevine. PMID:22702718

Evaluation of blotchy pigments in the anterior chamber angle as a sign of angle closure

PubMed Central

Rao, Harsha L; Mungale, Sachin C; Kumbar, Tukaram; Parikh, Rajul S; Garudadri, Chandra S

2012-01-01

Background: Blotchy pigments in the anterior chamber (AC) angle are considered diagnostic of primary angle closure (PAC). But there are no reports either on the prevalence of blotchy pigments in AC angles or the validity of this sign. Aims: To determine the prevalence of blotchy pigments in AC angles and to evaluate their relationship with glaucomatous optic neuropathy (GON) in eyes with occludable angles. Setting and Design: Cross-sectional, comparative study. Materials and Methods: Gonioscopy was performed in 1001 eyes of 526 subjects (245 eyes of 148 consecutive, occludable angle subjects and 756 eyes of 378 non-consecutive, open angle subjects), above 35 years of age. Quadrant-wise location of blotchy pigments was documented. Statistical Analysis: Odds of blotchy pigments in occludable angles against that in open angles were evaluated. Relationship of GON with blotchy pigments in occludable angle eyes was evaluated using a multivariate model. Results: Prevalence of blotchy pigments in occludable angles was 28.6% (95% CI, 22.9-34.3) and in open angles was 4.7% (95% CI, 3.2-6.3). Blotchy pigments were more frequently seen in inferior (16%) and superior quadrants (15%) of occludable angles, and inferior quadrant of open angles (4%). Odds of superior quadrant blotchy pigments in occludable angles were 33 times that in open angles. GON was seen in 107 occludable angle eyes. Blotchy pigments were not significantly associated with GON (odds ratio = 0.5; P = 0.1). Conclusions: Blotchy pigments were seen in 28.6% of occludable angle eyes and 4.7% of open angles eyes. Presence of blotchy pigments in the superior quadrant is more common in occludable angles. Presence of GON in occludable angle eyes was not associated with blotchy pigments. PMID:23202393

Development of high performance liquid chromatography method for miconazole analysis in powder sample

NASA Astrophysics Data System (ADS)

Hermawan, D.; Suwandri; Sulaeman, U.; Istiqomah, A.; Aboul-Enein, H. Y.

2017-02-01

A simple high performance liquid chromatography (HPLC) method has been developed in this study for the analysis of miconazole, an antifungal drug, in powder sample. The optimized HPLC system using C8 column was achieved using mobile phase composition containing methanol:water (85:15, v/v), a flow rate of 0.8 mL/min, and UV detection at 220 nm. The calibration graph was linear in the range from 10 to 50 mg/L with r 2 of 0.9983. The limit of detection (LOD) and limit of quantitation (LOQ) obtained were 2.24 mg/L and 7.47 mg/L, respectively. The present HPLC method is applicable for the determination of miconazole in the powder sample with a recovery of 101.28 % (RSD = 0.96%, n = 3). The developed HPLC method provides short analysis time, high reproducibility and high sensitivity.

Leukotriene B4 catabolism: quantitation of leukotriene B4 and its omega-oxidation products by reversed-phase high-performance liquid chromatography.

PubMed

Shak, S

1987-01-01

LTB4 and its omega-oxidation products may be rapidly, sensitively, and specifically quantitated by the methods of solid-phase extraction and reversed-phase high-performance liquid chromatography (HPLC), which are described in this chapter. Although other techniques, such as radioimmunoassay or gas chromatography-mass spectrometry, may be utilized for quantitative analysis of the lipoxygenase products of arachidonic acid, only the technique of reversed-phase HPLC can quantitate as many as 10 metabolites in a single analysis, without prior derivatization. In this chapter, we also reviewed the chromatographic theory which we utilized in order to optimize reversed-phase HPLC analysis of LTB4 and its omega-oxidation products. With this information and a gradient HPLC system, it is possible for any investigator to develop a powerful assay for the potent inflammatory mediator, LTB4, or for any other lipoxygenase product of arachidonic acid.

Rapid NMR method for the quantification of organic compounds in thin stillage.

PubMed

Ratanapariyanuch, Kornsulee; Shen, Jianheng; Jia, Yunhua; Tyler, Robert T; Shim, Youn Young; Reaney, Martin J T

2011-10-12

Thin stillage contains organic and inorganic compounds, some of which may be valuable fermentation coproducts. This study describes a thorough analysis of the major solutes present in thin stillage as revealed by NMR and HPLC. The concentration of charged and neutral organic compounds in thin stillage was determined by excitation sculpting NMR methods (double pulse field gradient spin echo). Compounds identified by NMR included isopropanol, ethanol, lactic acid, 1,3-propanediol, acetic acid, succinic acid, glycerophosphorylcholine, betaine, glycerol, and 2-phenylethanol. The concentrations of lactic and acetic acid determined with NMR were comparable to those determined using HPLC. HPLC and NMR were complementary, as more compounds were identified using both methods. NMR analysis revealed that stillage contained the nitrogenous organic compounds betaine and glycerophosphorylcholine, which contributed as much as 24% of the nitrogen present in the stillage. These compounds were not observed by HPLC analysis.

Lack of tumour pigmentation in conjunctival melanoma is associated with light iris colour and worse prognosis.

PubMed

Brouwer, Niels J; Marinkovic, Marina; Luyten, Gregorius P M; Shields, Carol L; Jager, Martine J

2018-05-18

To investigate whether differences in iris colour, skin colour and tumour pigmentation are related to clinical outcome in conjunctival melanoma. Data of 70 patients with conjunctival melanoma from the Leiden University Medical Center (Leiden, The Netherlands) and 374 patients from the Wills Eye Hospital (Philadephia, USA) were reviewed. The relation between iris colour, skin colour and tumour pigmentation versus clinical parameters and outcome was investigated using univariate and multivariate regression analyses. A light iris colour (blue, grey, green) was present in 261 (59%) patients and a dark colour (hazel, brown) in 183 (41%). A low tumour pigmentation was detected in 130 (40%) and a high pigmentation in 197 (60%) patients. Low tumour pigmentation was associated with light iris colour (p=0.021) but not related to skin colour (p=0.92). In univariate analysis, neither iris nor skin colour was related to clinical outcome, while a low tumour pigmentation was related to metastasis formation (HR 2.37, p=0.004) and death (HR 2.42, p=0.020). In multivariate analysis, low tumour pigmentation was related to the development of recurrences (HR 1.63, p=0.043), metastasis formation (HR 2.48, p=0.004) and death (HR 2.60, p=0.014). Lightly pigmented tumours occurred especially in individuals with lightly coloured irises. While iris colour or skin colour was not significantly related to clinical outcome, a low tumour pigmentation was related to a worse outcome in patients with conjunctival melanoma. The amount and type of melanin in conjunctival melanocytes may be involved in the pathogenesis and behaviour of selected conjunctival melanoma. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

A molecular systems approach to modelling human skin pigmentation: identifying underlying pathways and critical components.

PubMed

Raghunath, Arathi; Sambarey, Awanti; Sharma, Neha; Mahadevan, Usha; Chandra, Nagasuma

2015-04-29

Ultraviolet radiations (UV) serve as an environmental stress for human skin, and result in melanogenesis, with the pigment melanin having protective effects against UV induced damage. This involves a dynamic and complex regulation of various biological processes that results in the expression of melanin in the outer most layers of the epidermis, where it can exert its protective effect. A comprehensive understanding of the underlying cross talk among different signalling molecules and cell types is only possible through a systems perspective. Increasing incidences of both melanoma and non-melanoma skin cancers necessitate the need to better comprehend UV mediated effects on skin pigmentation at a systems level, so as to ultimately evolve knowledge-based strategies for efficient protection and prevention of skin diseases. A network model for UV-mediated skin pigmentation in the epidermis was constructed and subjected to shortest path analysis. Virtual knock-outs were carried out to identify essential signalling components. We describe a network model for UV-mediated skin pigmentation in the epidermis. The model consists of 265 components (nodes) and 429 directed interactions among them, capturing the manner in which one component influences the other and channels information. Through shortest path analysis, we identify novel signalling pathways relevant to pigmentation. Virtual knock-outs or perturbations of specific nodes in the network have led to the identification of alternate modes of signalling as well as enabled determining essential nodes in the process. The model presented provides a comprehensive picture of UV mediated signalling manifesting in human skin pigmentation. A systems perspective helps provide a holistic purview of interconnections and complexity in the processes leading to pigmentation. The model described here is extensive yet amenable to expansion as new data is gathered. Through this study, we provide a list of important proteins essential for pigmentation which can be further explored to better understand normal pigmentation as well as its pathologies including vitiligo and melanoma, and enable therapeutic intervention.

[Cutaneous pigmentation related to intravenous iron extravasation: Analysis from the French pharmacovigilance database].

PubMed

Hermitte-Gandoliere, Alexia; Petitpain, Nadine; Lepelley, Marion; Thomas, Laure; Le Beller, Christine; Astoul, Jacqueline Ponte; Gillet, Pierre

Intravenous iron infusion may be complicated by extravasation and lead to cutaneous pigmentation. We queried the French pharmacovigilance database to assess the spontaneously reported cases over the 2000-2016 period. Fifty-one cases of cutaneous pigmentation related to intravenous iron extravasation were retrieved, none was associated to necrosis. Most of patients were women aged 20 to 49 years old. The pigmentation was mostly a brown coloration, persisting over one month in 19 cases (37.2%) and over 6 months in 9 cases (17.6%). The management of extravasation and pigmentation was heterogeneous and was rarely followed by a decrease of the coloration. Cutaneous pigmentation after intravenous iron extravasation can persist over time and create an aesthetic prejudice, particularly in young women. Standardized extravasation and iron-induced pigmentation management procedures appear necessary. Copyright © 2017 Société française de pharmacologie et de thérapeutique. Published by Elsevier Masson SAS. All rights reserved.

Portable laser-induced breakdown spectroscopy/diffuse reflectance hybrid spectrometer for analysis of inorganic pigments

NASA Astrophysics Data System (ADS)

Siozos, Panagiotis; Philippidis, Aggelos; Anglos, Demetrios

2017-11-01

A novel, portable spectrometer, combining two analytical techniques, laser-induced breakdown spectroscopy (LIBS) and diffuse reflectance spectroscopy, was developed with the aim to provide an enhanced instrumental and methodological approach with regard to the analysis of pigments in objects of cultural heritage. Technical details about the hybrid spectrometer and its operation are presented and examples are given relevant to the analysis of paint materials. Both LIBS and diffuse reflectance spectra in the visible and part of the near infrared, corresponding to several neat mineral pigment samples, were recorded and the complementary information was used to effectively distinguish different types of pigments even if they had similar colour or elemental composition. The spectrometer was also employed in the analysis of different paints on the surface of an ancient pottery sherd demonstrating the capabilities of the proposed hybrid diagnostic approach. Despite its instrumental simplicity and compact size, the spectrometer is capable of supporting analytical campaigns relevant to archaeological, historical or art historical investigations, particularly when quick data acquisition is required in the context of surveys of large numbers of objects and samples.

Quantification of early cutaneous manifestations of chronic venous insufficiency by automated analysis of photographic images: Feasibility and technical considerations.

PubMed

Becker, François; Fourgeau, Patrice; Carpentier, Patrick H; Ouchène, Amina

2018-06-01

We postulate that blue telangiectasia and brownish pigmentation at ankle level, early markers of chronic venous insufficiency, can be quantified for longitudinal studies of chronic venous disease in Caucasian people. Objectives and methods To describe a photographic technique specially developed for this purpose. The pictures were acquired using a dedicated photo stand to position the foot in a reproducible way, with a normalized lighting and acquisition protocol. The image analysis was performed with a tool developed using algorithms optimized to detect and quantify blue telangiectasia and brownish pigmentation and their relative surface in the region of interest. To test the short-term reproducibility of the measures. Results The quantification of the blue telangiectasia and of the brownish pigmentation using an automated digital photo analysis is feasible. The short-term reproducibility is good for blue telangiectasia quantification. It is a less accurate for the brownish pigmentation. Conclusion The blue telangiectasia of the corona phlebectatica and the ankle flare can be assessed using a clinimetric approach based on the automated digital photo analysis.

Independent component analysis-based algorithm for automatic identification of Raman spectra applied to artistic pigments and pigment mixtures.

PubMed

González-Vidal, Juan José; Pérez-Pueyo, Rosanna; Soneira, María José; Ruiz-Moreno, Sergio

2015-03-01

A new method has been developed to automatically identify Raman spectra, whether they correspond to single- or multicomponent spectra. The method requires no user input or judgment. There are thus no parameters to be tweaked. Furthermore, it provides a reliability factor on the resulting identification, with the aim of becoming a useful support tool for the analyst in the decision-making process. The method relies on the multivariate techniques of principal component analysis (PCA) and independent component analysis (ICA), and on some metrics. It has been developed for the application of automated spectral analysis, where the analyzed spectrum is provided by a spectrometer that has no previous knowledge of the analyzed sample, meaning that the number of components in the sample is unknown. We describe the details of this method and demonstrate its efficiency by identifying both simulated spectra and real spectra. The method has been applied to artistic pigment identification. The reliable and consistent results that were obtained make the methodology a helpful tool suitable for the identification of pigments in artwork or in paint in general.

[Simultaneous determination of 6 forbidden colorants in cosmetics by high performance liquid chromatography-tandem mass spectrometry].

PubMed

Lin, Weixuan; Sun, Xingquan; Zhao, Xuerong; Xu, Wei; Guo, Guiyuan

2012-05-01

A method of high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) has been established for the simultaneous determination of six forbidden colorants including Sudan IV, Acid Violet 49, Sudan Blue 2, Solvent Red 49, Basic Violet 1 and Pigment Orange 5 in cream and powdery matrix cosmetics. The samples were extracted with ethanol-acetonitrile (3:2, v/v) solution by ultrasonic technique for 20 min, then centrifuged for purification and enriched by nitrogen blowing sequentially. The analytes were isolated on a Luna C18 column (150 mm x 2.1 mm, 5 microm) by gradient elution with methanol and 10 mmol/L ammonium acetate as the mobile phases, and detected by MS/MS in the multiple reaction monitoring (MRM) mode. The qualitative analysis was based on the retention time and the relative abundance ratio of the characteristic ions, and the quantitative analysis on calibration curve method. The results showed that the limits of quantification (LOQ, S/N= 10) of the six colorants ranged from 0.1 to 10 microg/kg and the average recoveries were from 86.67% to 98.22% with the relative standard deviations (RSDs) from 4.01% to 7.01%. The method is simple and rapid with high sensitivity and good reproducibility, and suitable for the determination of the six forbidden colorants in cosmetics.

Rapid, accurate, and direct determination of total lycopene content in tomato paste

NASA Astrophysics Data System (ADS)

Bicanic, D.; Anese, M.; Luterotti, S.; Dadarlat, D.; Gibkes, J.; Lubbers, M.

2003-01-01

Lycopene that imparts red color to the tomato fruit is the most potent antioxidant among carotenes, an important nutrient and also used as a color ingredient in many food formulations. Since cooked and processed foods derived from tomatoes were shown to provide optimal lycopene boost, products such as paste, puree, juice, etc. are nowadays gaining popularity as dietary sources. The analysis of lycopene in tomato paste (partially dehydrated product prepared by vacuum concentrating tomato juice) is carried out using either high pressure liquid chromatography (HPLC), spectrophotometry, or by evaluating the color. The instability of lycopene during processes of extraction, etc., handling, and disposal of organic solvents makes the preparation of a sample for the analysis a delicate task. Despite a recognized need for accurate and rapid assessment of lycopene in tomato products no such method is available at present. The study described here focuses on a direct determination of a total lycopene content in different tomato pastes by means of the laser optothermal window (LOW) method at 502 nm. The concentration of lycopene in tomato paste ranged between 25 and 150 mg per 100 g product; the results are in excellent agreement with those obtained by spectrophotometry. The time needed to complete LOW analysis is very short, so that decomposition of pigment and the formation of artifacts are minimized. Preliminary results indicate a good degree of reproducibility making the LOW method suitable for routine assays of lycopene content in tomato paste.

X-ray fluorescence analysis of yellow pigments in altarpieces by Valencian artists of the XV and XVI centuries

NASA Astrophysics Data System (ADS)

Ferrero, J. L.; Roldán, C.; Ardid, M.; Navarro, E.

1999-02-01

XRF analysis has allowed a quick and precise detection and identification of the inorganic elements that compose the yellow pigments in altarpieces of the XV and XVI centuries painted by the Valencian artists Miguel Alcañiz, Vicente Macip, Juan de Juanes, Hernando Yáñez de la Almedina and Hernando Llanos. The analyses have been carried out with an XRF portable system that consists of a tube of X-rays and detectors of Si(Li) and cadmium zinc telluride. This system has enabled a non-aggressive and non-destructive analysis of many pieces at the Museo de Bellas Artes of Valencia (Spain). Among the yellow pigments we have identified a pigment composed by lead and tin oxides named lead-tin yellow (Pb 2SnO 4), frequently used in European paintings from the XIV century until the first half of the XVIII century. This fact demonstrates the influence of elements and pictorial techniques from Europe to the region of Valencia.

Lipid analysis via HPLC with a charged aerosol detector

USDA-ARS?s Scientific Manuscript database

Most lipid extracts are a mixture of saturated and unsaturated molecules. Therefore, the most successful HPLC detectors for the quantitative analysis of lipids have involved the use of “universal” or “mass” detectors such as flame ionization detectors (FID) and evaporative light scattering detectors...

An extended chemical analysis of gallstone.

PubMed

Chandran, P; Kuchhal, N K; Garg, P; Pundir, C S

2007-09-01

Chemical composition of gall stones is essential for aetiopathogensis of gallstone disease. We have reported quantitative chemical analysis of total cholesterol bilirubin, calcium, iron and inorganic phosphate in 120 gallstones from haryana. To extend this chemical analysis of gall stones by studying more cases and by analyzing more chemical constituents. A quantitative chemical analysis of total cholesterol, total bilirubin, fatty acids, triglycerides, phospholipids, bile acids, soluble proteins, sodium potassium, magnesium, copper, oxalate and chlorides of biliary calculi (52 cholesterol, 76 mixed and 72 pigment) retrieved from surgical operation of 200 patients from Haryana state was carried out. Total cholesterol as the major component and total bilirubin, phospholipids, triglycerides, bile acids, fatty acids (esterified), soluble protein, calcium, magnesium, iron, copper, sodium, potassium, inorganic phosphate, oxalate and chloride as minor components were found in all types of calculi. The cholesterol stones had higher content of total cholesterol, phospholipids, fatty acids (esterified), inorganic phosphate and copper compared to mixed and pigment stones. The mixed stones had higher content of iron and triglycerides than to cholesterol and pigment stones. The pigment stones were richer in total bilirubin, bile acids, calcium, oxalate, magnesium, sodium, potassium, chloride and soluble protein compared to cholesterol and mixed stones. Although total cholesterol was a major component of cholesterol, mixed and pigment gall stone in Haryana, the content of most of the other lipids, cations and anions was different in different gall stones indicating their different mechanism of formation.

Screening and Analysis of the Marker Components in Ganoderma lucidum by HPLC and HPLC-MSn with the Aid of Chemometrics.

PubMed

Wu, Lingfang; Liang, Wenyi; Chen, Wenjing; Li, Shi; Cui, Yaping; Qi, Qi; Zhang, Lanzhen

2017-04-06

Ganoderma triterpenes (GTs) are the major secondary metabolites of Ganoderma lucidum , which is a popularly used traditional Chinese medicine for complementary cancer therapy. The present study was to establish a fingerprint evaluation system based on Similarity Analysis (SA), Cluster Analysis (CA) and Principal Component Analysis (PCA) for the identification and quality control of G. lucidum . Fifteen samples from the Chinese provinces of Hainan, Neimeng, Shangdong, Jilin, Anhui, Henan, Yunnan, Guangxi and Fujian were analyzed by HPLC-PAD and HPLC-MS n . Forty-seven compounds were detected by HPLC, of which forty-two compounds were tentatively identified by comparing their retention times and mass spectrometry data with that of reference compounds and reviewing the literature. Ganoderic acid B, 3,7,15-trihydroxy-11,23-dioxolanost-8,16-dien-26-oic acid, lucidenic acid A, ganoderic acid G, and 3,7-oxo-12-acetylganoderic acid DM were deemed to be the marker compounds to distinguish the samples with different quality according to both CA and PCA. This study provides helpful chemical information for further research on the anti-tumor activity and mechanism of action of G. lucidum . The results proved that fingerprints combined with chemometrics are a simple, rapid and effective method for the quality control of G. lucidum .

Ellagitannin composition of blackberry as determined by HPLC-ESI-MS and MALDI-TOF-MS.

PubMed

Hager, Tiffany J; Howard, Luke R; Liyanage, Rohana; Lay, Jackson O; Prior, Ronald L

2008-02-13

Blackberries ( Rubus sp.) were evaluated by high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) to identify the ellagitannins present in flesh, torus (receptacle tissue), and seeds. Most ellagitannins were present (or detectable) only in seed tissues. Ellagitannins identified by HPLC-ESI-MS in the seeds included pedunculagin, casuarictin/potentillin, castalagin/vescalagin, lambertianin A/sanguiin H-6, lambertianin C, and lambertianin D. For several of the ellagitannins, isomeric separation was also obtained. The MALDI-TOF-MS analysis was primarily utilized to evaluate and identify high molecular mass (>1000 Da) ellagitannins. The MALDI analysis verified the presence of the ellagitannins identified by HPLC-ESI-MS including lambertianin A/sanguiin H-6, lambertianin C, and lambertianin D, but the analysis also indicated the presence of several other compounds that were most likely ellagitannins based on the patterns observed in the masses (i.e., loss or addition of a gallic acid moiety to a known ellagitannin). This study determined the presence of several possible isomeric forms of ellagitannins previously unidentified in fruit and presents a possible analytical HPLC method for the analysis of the major ellagitannins present in the fruit.

X-ray texture analysis of paper coating pigments and the correlation with chemical composition analysis

NASA Astrophysics Data System (ADS)

Roine, J.; Tenho, M.; Murtomaa, M.; Lehto, V.-P.; Kansanaho, R.

2007-10-01

The present research experiments the applicability of x-ray texture analysis in investigating the properties of paper coatings. The preferred orientations of kaolin, talc, ground calcium carbonate, and precipitated calcium carbonate particles used in four different paper coatings were determined qualitatively based on the measured crystal orientation data. The extent of the orientation, namely, the degree of the texture of each pigment, was characterized quantitatively using a single parameter. As a result, the effect of paper calendering is clearly seen as an increase on the degree of texture of the coating pigments. The effect of calendering on the preferred orientation of kaolin was also evident in an independent energy dispersive spectrometer analysis on micrometer scale and an electron spectroscopy for chemical analysis on nanometer scale. Thus, the present work proves x-ray texture analysis to be a potential research tool for characterizing the properties of paper coating layers.

[Study on the fingerprint of Morus alba from different habitats by HPLC].

PubMed

Chen, Cheng; Li, Hong-Bo; Wang, Liu-Ping; Li, Yun-Rong; Xin, Ning

2012-12-01

To establish HPLC fingerprint of Morus alba from different habitats by HPLC and provide basis for its quality control. HPLC analysis was performed on an Agilent XDB C18 Column (4.6 mm x 250 mm, 5 microm), gradient eluted composed of acetonitrile and 0.3% phosphate acid. The column temperature was set at 35 degrees C and the flow rate was 0.5 mL/min. The detective wavelength was 290 nm. The HPLC fingerprint for 10 batches of Morus alba was studied on their similarity. There were twelve common peaks in the fingerprint. The similarity of 7 batches was above 0.9 and the other batches had low similarity. The HPLC fingerprint can be used for quality control of Morus alba with high characteristics and specificity.

Genetic, epigenetic, and HPLC fingerprint differentiation between natural and ex situ populations of Rhodiola sachalinensis from Changbai Mountain, China.

PubMed

Zhao, Wei; Shi, Xiaozheng; Li, Jiangnan; Guo, Wei; Liu, Chengbai; Chen, Xia

2014-01-01

Rhodiola sachalinensis is an endangered species with important medicinal value. We used inter-simple sequence repeat (ISSR) and methylation-sensitive amplified polymorphism (MSAP) markers to analyze genetic and epigenetic differentiation in different populations of R. sachalinensis, including three natural populations and an ex situ population. Chromatographic fingerprint was used to reveal HPLC fingerprint differentiation. According to our results, the ex situ population of R. sachalinensis has higher level genetic diversity and greater HPLC fingerprint variation than natural populations, but shows lower epigenetic diversity. Most genetic variation (54.88%) was found to be distributed within populations, and epigenetic variation was primarily distributed among populations (63.87%). UPGMA cluster analysis of ISSR and MSAP data showed identical results, with individuals from each given population grouping together. The results of UPGMA cluster analysis of HPLC fingerprint patterns was significantly different from results obtained from ISSR and MSAP data. Correlation analysis revealed close relationships among altitude, genetic structure, epigenetic structure, and HPLC fingerprint patterns (R2 = 0.98 for genetic and epigenetic distance; R2 = 0.90 for DNA methylation level and altitude; R2 = -0.95 for HPLC fingerprint and altitude). Taken together, our results indicate that ex situ population of R. sachalinensis show significantly different genetic and epigenetic population structures and HPLC fingerprint patterns. Along with other potential explanations, these findings suggest that the ex situ environmental factors caused by different altitude play an important role in keeping hereditary characteristic of R. sachalinensis.

Comparison of a fluorometric assay kit with high-performance liquid chromatography for the assessment of serum retinol concentration.

PubMed

Elom, Aglago Kouassivi; Imane, El Menchawy; Kaoutar, Benjeddou; Khalid, El Kari; Asmaa, El Hamdouchi; Mehdi, Azlaf; Noureddine, El Haloui; Hassan, Aguenaou

2015-06-01

Although high-performance liquid chromatography (HPLC) is the commonly used method for the analysis of retinol in biological samples, simple and rapid test kits are available. This study compared a rapid test kit (ICHECK Fluoro®) to HPLC for the assessment of serum retinol concentrations. For the analysis by HPLC, sample preparation included standard deproteinization and extraction phases. The analysis by ICHECK was performed by injecting serum into IEX reagent vials (n=89) and mixing manually for separation. After precipitation of the proteins, the vial was introduced into the chamber of the ICHECK Fluoro and analysed at 0 min (ICHECK0min) and 15 min later (ICHECK15min). Bland and Altman approach was applied to test the agreement between HPLC and ICHECK. Mean HPLC, ICHECK0min and ICHECK15min values were 421.2±106.0 µg/L, 423.1±118.3 µg/L and 413.2±107.6 µg/L, respectively. Retinol concentrations significantly decreased in the IEX solution over time (p<0.001). No significant proportional bias was observed between HPLC and ICHECK0min (r-0.038, p=0.73) and ICHECK15min (r=-0.024, p=0.82). Fixed biases (HPLC minus ICHECK) for ICHECK0min and ICHECK15min were respectively -1.9±23.1 µg/l (p=0.45) and 8.0±22.7 µg/l (p=0.002). ICHECK Fluoro may offer a reliable mean for assessing serum retinol for measurements performed with no significant time delay.

Genetic, Epigenetic, and HPLC Fingerprint Differentiation between Natural and Ex Situ Populations of Rhodiola sachalinensis from Changbai Mountain, China

PubMed Central

Zhao, Wei; Shi, Xiaozheng; Li, Jiangnan; Guo, Wei; Liu, Chengbai; Chen, Xia

2014-01-01

Rhodiola sachalinensis is an endangered species with important medicinal value. We used inter-simple sequence repeat (ISSR) and methylation-sensitive amplified polymorphism (MSAP) markers to analyze genetic and epigenetic differentiation in different populations of R. sachalinensis, including three natural populations and an ex situ population. Chromatographic fingerprint was used to reveal HPLC fingerprint differentiation. According to our results, the ex situ population of R. sachalinensis has higher level genetic diversity and greater HPLC fingerprint variation than natural populations, but shows lower epigenetic diversity. Most genetic variation (54.88%) was found to be distributed within populations, and epigenetic variation was primarily distributed among populations (63.87%). UPGMA cluster analysis of ISSR and MSAP data showed identical results, with individuals from each given population grouping together. The results of UPGMA cluster analysis of HPLC fingerprint patterns was significantly different from results obtained from ISSR and MSAP data. Correlation analysis revealed close relationships among altitude, genetic structure, epigenetic structure, and HPLC fingerprint patterns (R2 = 0.98 for genetic and epigenetic distance; R2 = 0.90 for DNA methylation level and altitude; R2 = –0.95 for HPLC fingerprint and altitude). Taken together, our results indicate that ex situ population of R. sachalinensis show significantly different genetic and epigenetic population structures and HPLC fingerprint patterns. Along with other potential explanations, these findings suggest that the ex situ environmental factors caused by different altitude play an important role in keeping hereditary characteristic of R. sachalinensis. PMID:25386983

Melanocyte pigmentation stiffens murine cardiac tricuspid valve leaflet

PubMed Central

Balani, Kantesh; Brito, Flavia C.; Kos, Lidia; Agarwal, Arvind

2009-01-01

Pigmentation of murine cardiac tricuspid valve leaflet is associated with melanocyte concentration, which affects its stiffness. Owing to its biological and viscoelastic nature, estimation of the in situ stiffness measurement becomes a challenging task. Therefore, quasi-static and nanodynamic mechanical analysis of the leaflets of the mouse tricuspid valve is performed in the current work. The mechanical properties along the leaflet vary with the degree of pigmentation. Pigmented regions of the valve leaflet that contain melanocytes displayed higher storage modulus (7–10 GPa) than non-pigmented areas (2.5–4 GPa). These results suggest that the presence of melanocytes affects the viscoelastic properties of the mouse atrioventricular valves and are important for their proper functioning in the organism. PMID:19586956

Changes in pigment, spectral transmission and element content of pink chicken eggshells with different pigment intensity during incubation.

PubMed

Yu, Yue; Li, Zhanming; Pan, Jinming

2016-01-01

Objective. The objective of this study was to investigate changes in pigment, spectral transmission and element content of chicken eggshells with different intensities of pink pigment during the incubation period. We also investigated the effects of the region (small pole, equator and large pole) and pink pigment intensity of the chicken eggshell on the percent transmission of light passing through the chicken eggshells. Method. Eggs of comparable weight from a meat-type breeder (Meihuang) were used, and divided based on three levels of pink pigment (light, medium and dark) in the eggshells. During the incubation (0-21 d), the values of the eggshell pigment (ΔE, L (∗), a (∗), b (∗)) were measured. The percent transmission of light for different regions and intensities of eggshell pigmentation was measured by using the visible wavelength range of 380-780 nm. Result. Three measured indicators of eggshell color, ΔE, L (∗) and a (∗), did not change significantly during incubation. Compared with other regions and pigment intensities, eggshell at the small pole and with light pigmentation intensity showed the highest percent transmission of light. The transmission value varied significantly (P < 0.001) with incubation time. The element analysis of eggshells with different levels of pink pigment showed that the potassium content of the eggshells for all pigment levels decreased significantly during incubation. Conclusion. In summary, pigment intensity and the region of the eggshell influenced the percent transmission of light of eggshell. Differences in the spectral characteristics of different eggshells may influence the effects of photostimulation during the incubation of eggs. All of these results will be applicable for perfecting the design of light intensity for lighted incubation to improve productivity.

Changes in pigment, spectral transmission and element content of pink chicken eggshells with different pigment intensity during incubation

PubMed Central

Yu, Yue; Li, Zhanming

2016-01-01

Objective. The objective of this study was to investigate changes in pigment, spectral transmission and element content of chicken eggshells with different intensities of pink pigment during the incubation period. We also investigated the effects of the region (small pole, equator and large pole) and pink pigment intensity of the chicken eggshell on the percent transmission of light passing through the chicken eggshells. Method. Eggs of comparable weight from a meat-type breeder (Meihuang) were used, and divided based on three levels of pink pigment (light, medium and dark) in the eggshells. During the incubation (0–21 d), the values of the eggshell pigment (ΔE, L∗, a∗, b∗) were measured. The percent transmission of light for different regions and intensities of eggshell pigmentation was measured by using the visible wavelength range of 380–780 nm. Result. Three measured indicators of eggshell color, ΔE, L∗ and a∗, did not change significantly during incubation. Compared with other regions and pigment intensities, eggshell at the small pole and with light pigmentation intensity showed the highest percent transmission of light. The transmission value varied significantly (P < 0.001) with incubation time. The element analysis of eggshells with different levels of pink pigment showed that the potassium content of the eggshells for all pigment levels decreased significantly during incubation. Conclusion. In summary, pigment intensity and the region of the eggshell influenced the percent transmission of light of eggshell. Differences in the spectral characteristics of different eggshells may influence the effects of photostimulation during the incubation of eggs. All of these results will be applicable for perfecting the design of light intensity for lighted incubation to improve productivity. PMID:27019785

Analysis of pigments in polychromes by use of laser induced breakdown spectroscopy and Raman microscopy

NASA Astrophysics Data System (ADS)

Castillejo, M.; Martín, M.; Silva, D.; Stratoudaki, T.; Anglos, D.; Burgio, L.; Clark, R. J. H.

2000-09-01

Two laser-based analytical techniques, Laser Induced Breakdown Spectroscopy (LIBS) and Raman microscopy, have been used for the identification of pigments on a polychrome from the Rococo period. Detailed spectral data are presented from analyses performed on a fragment of a gilded altarpiece from the church of Escatrón, Zaragoza, Spain. LIBS measurements yielded elemental analytical data which suggest the presence of certain pigments and, in addition, provide information on the stratigraphy of the paint layers. Identification of most pigments and of the materials used in the preparation layer was performed by Raman microscopy.

HPLC Analysis and Cytotoxicity of n-Butanol Extract from Glyphaea brevis Roots Against C6 Glioma Cells.

PubMed

Konan, Koffi Marcel; Mamyrbekova-Bekro, Janat Akhanovna; Bakalara, Norbert; Virieux, David; Pirat, Jean-Luc; Bekro, Yves-Alain

2014-01-01

The n-butanol extract of the roots of Glyphaea brevis was analysed. HPLC analysis suggested the presence of phenolic compounds like protocatechuic acid (PCA). The extract showed moderate cytotoxic activity against C6 glioma cells (EC50 > 1 mg/ml).

HPLC Analysis and Cytotoxicity of n-Butanol Extract from Glyphaea brevis Roots Against C6 Glioma Cells

PubMed Central

Konan, Koffi Marcel; Mamyrbekova-Bekro, Janat Akhanovna; Bakalara, Norbert; Virieux, David; Pirat, Jean-Luc; Bekro, Yves-Alain

2014-01-01

The n-butanol extract of the roots of Glyphaea brevis was analysed. HPLC analysis suggested the presence of phenolic compounds like protocatechuic acid (PCA). The extract showed moderate cytotoxic activity against C6 glioma cells (EC50 > 1 mg/ml). PMID:24634849

Adaptation of cone pigments found in green rods for scotopic vision through a single amino acid mutation

PubMed Central

Kojima, Keiichi; Matsutani, Yuki; Yamashita, Takahiro; Yanagawa, Masataka; Imamoto, Yasushi; Yamano, Yumiko; Wada, Akimori; Hisatomi, Osamu; Nishikawa, Kanto; Sakurai, Keisuke; Shichida, Yoshinori

2017-01-01

Most vertebrate retinas contain a single type of rod for scotopic vision and multiple types of cones for photopic and color vision. The retinas of certain amphibian species uniquely contain two types of rods: red rods, which express rhodopsin, and green rods, which express a blue-sensitive cone pigment (M1/SWS2 group). Spontaneous activation of rhodopsin induced by thermal isomerization of the retinal chromophore has been suggested to contribute to the rod’s background noise, which limits the visual threshold for scotopic vision. Therefore, rhodopsin must exhibit low thermal isomerization rate compared with cone visual pigments to adapt to scotopic condition. In this study, we determined whether amphibian blue-sensitive cone pigments in green rods exhibit low thermal isomerization rates to act as rhodopsin-like pigments for scotopic vision. Anura blue-sensitive cone pigments exhibit low thermal isomerization rates similar to rhodopsin, whereas Urodela pigments exhibit high rates like other vertebrate cone pigments present in cones. Furthermore, by mutational analysis, we identified a key amino acid residue, Thr47, that is responsible for the low thermal isomerization rates of Anura blue-sensitive cone pigments. These results strongly suggest that, through this mutation, anurans acquired special blue-sensitive cone pigments in their green rods, which could form the molecular basis for scotopic color vision with normal red rods containing green-sensitive rhodopsin. PMID:28484015

Raman spectroscopic analysis of a 'noli me tangere' painting.

PubMed

Hibberts, Stephen; Edwards, Howell G M; Abdel-Ghani, Mona; Vandenabeele, Peter

2016-12-13

The discovery of an oil painting in seriously damaged condition with an important historical and a heterodox detail with possible origins in the late fifteenth century has afforded the opportunity for Raman microscopic analysis prior to its restoration being undertaken. The painting depicts a risen Christ following His crucifixion in a 'noli me tangere' pose with three women in an Italian terrace garden with a stone balustrade overlooking a rural landscape and an undoubted view of late-medieval Florence. The picture has suffered much abuse and is in very poor condition, which is possibly attributable to its controversial portrayal of a polydactylic Christ with six toes on His right foot. By the late sixteenth century, after the Council of Trent, this portrayal would almost certainly have been frowned upon by the Church authorities or more controversially as a depiction of the holy. Raman spectroscopic analysis of the pigments places the painting as being consistent chronologically with the Renaissance period following the identification of cinnabar, haematite, red lead, lead white, goethite, verdigris, caput mortuum and azurite with no evidence of more modern synthetic pigments or of modern restoration having been carried out. An interesting pigment mixture found here is that of the organic dye carmine and cinnabar to produce a particular bright red pigment coloration. Stratigraphic examination of the paint fragments has demonstrated the presence of an orange resin layer immediately on top of the canvas substrate, effectively rendering the pigment as a sandwich between this substratal resin and the overlying varnish. The Raman spectroscopic evidence clearly indicates that an attribution of the artwork to the Renaissance is consistent with the scientific analysis of the pigment composition.This article is part of the themed issue 'Raman spectroscopy in art and archaeology'. © 2016 The Author(s).

Raman spectroscopic analysis of a `noli me tangere' painting

NASA Astrophysics Data System (ADS)

Hibberts, Stephen; Edwards, Howell G. M.; Abdel-Ghani, Mona; Vandenabeele, Peter

2016-12-01

The discovery of an oil painting in seriously damaged condition with an important historical and a heterodox detail with possible origins in the late fifteenth century has afforded the opportunity for Raman microscopic analysis prior to its restoration being undertaken. The painting depicts a risen Christ following His crucifixion in a `noli me tangere' pose with three women in an Italian terrace garden with a stone balustrade overlooking a rural landscape and an undoubted view of late-medieval Florence. The picture has suffered much abuse and is in very poor condition, which is possibly attributable to its controversial portrayal of a polydactylic Christ with six toes on His right foot. By the late sixteenth century, after the Council of Trent, this portrayal would almost certainly have been frowned upon by the Church authorities or more controversially as a depiction of the holy. Raman spectroscopic analysis of the pigments places the painting as being consistent chronologically with the Renaissance period following the identification of cinnabar, haematite, red lead, lead white, goethite, verdigris, caput mortuum and azurite with no evidence of more modern synthetic pigments or of modern restoration having been carried out. An interesting pigment mixture found here is that of the organic dye carmine and cinnabar to produce a particular bright red pigment coloration. Stratigraphic examination of the paint fragments has demonstrated the presence of an orange resin layer immediately on top of the canvas substrate, effectively rendering the pigment as a sandwich between this substratal resin and the overlying varnish. The Raman spectroscopic evidence clearly indicates that an attribution of the artwork to the Renaissance is consistent with the scientific analysis of the pigment composition. This article is part of the themed issue "Raman spectroscopy in art and archaeology".

40 CFR 86.542-90 - Records required.

Code of Federal Regulations, 2013 CFR

2013-07-01

... formaldehyde sampling system. (6) The formaldehyde calibration information from the HPLC standards. (7) The concentration of the HPLC analysis of the test sample (formaldehyde). (q) Additional required records for...

40 CFR 86.542-90 - Records required.

Code of Federal Regulations, 2014 CFR

2014-07-01

... formaldehyde sampling system. (6) The formaldehyde calibration information from the HPLC standards. (7) The concentration of the HPLC analysis of the test sample (formaldehyde). (q) Additional required records for...

40 CFR 86.542-90 - Records required.

Code of Federal Regulations, 2011 CFR

2011-07-01

... formaldehyde sampling system. (6) The formaldehyde calibration information from the HPLC standards. (7) The concentration of the HPLC analysis of the test sample (formaldehyde). (q) Additional required records for...

40 CFR 86.542-90 - Records required.

Code of Federal Regulations, 2012 CFR

2012-07-01

... formaldehyde sampling system. (6) The formaldehyde calibration information from the HPLC standards. (7) The concentration of the HPLC analysis of the test sample (formaldehyde). (q) Additional required records for...

Quantitative X-ray diffraction and fluorescence analysis of paint pigment systems : final report.

DOT National Transportation Integrated Search

1978-01-01

This study attempted to correlate measured X-ray intensities with concentrations of each member of paint pigment systems, thereby establishing calibration curves for the quantitative analyses of such systems.

[Indices of pigment formation in complex cytomorphological research on human pigmented skin neoplasms].

PubMed

Naleskina, L A

1985-01-01

Analysis of the topography peculiarities and distribution of oxidized melanine and its precursors (DOPA-oxide activity and catecholamine) in pigment nevuses and malignant melanomas of skin shows that the studied peculiarities are a complex of intersupplementary markers of melanine formation, correlate with the quality and the degree of proliferative process expression in tumours of this genesis and may be used for their malignancy rating.

In situ X-ray fluorescence-based method to differentiate among red ochre pigments and yellow ochre pigments thermally transformed to red pigments of wall paintings from Pompeii.

PubMed

Marcaida, Iker; Maguregui, Maite; Fdez-Ortiz de Vallejuelo, Silvia; Morillas, Héctor; Prieto-Taboada, Nagore; Veneranda, Marco; Castro, Kepa; Madariaga, Juan Manuel

2017-06-01

Most of the magnificent wall paintings from the ancient city of Pompeii are decorated with red and yellow colors coming from the ochre pigments used. The thermal impact of the pyroclastic flow from the eruption of Vesuvius, in AD 79, promoted the transformation of some yellow painted areas to red. In this work, original red ochre, original yellow ochre, and transformed yellow ochre (nowadays showing a red color) of wall paintings from Pompeian houses (House of Marcus Lucretius and House of Gilded Cupids) were analyzed by means of a handheld energy-dispersive X-ray fluorescence spectrometer to develop a fast method that allows chemical differentiation of the original red ochre and the transformed yellow ochre. Principal component analysis of the multivariate obtained data showed that arsenic is the tracer element to distinguish between both red colored ochres. Moreover, Pompeian raw red and yellow ochre pigments recovered from the burial were analyzed in the laboratory with use of a benchtop energy-dispersive X-ray fluorescence spectrometer to confirm the elemental composition and the conclusions drawn from the in situ analysis according to the yellow ochre pigment transformation in real Pompeian wall paintings.

Stable Benzacridine Pigments by Oxidative Coupling of Chlorogenic Acid with Amino Acids and Proteins: Toward Natural Product-Based Green Food Coloring.

PubMed

Iacomino, Mariagrazia; Weber, Fabian; Gleichenhagen, Maike; Pistorio, Valeria; Panzella, Lucia; Pizzo, Elio; Schieber, Andreas; d'Ischia, Marco; Napolitano, Alessandra

2017-08-09

The occasional greening of sweet potatoes and other plant tissues observed during cooking or other food processing has been shown to arise from the autoxidative coupling of chlorogenic acid (CGA, 5-caffeoylquinic acid) with amino acid components, leading to trihydroxybenzacridine pigments. To explore the potential of this reaction for food coloring, we report herein the optimized biomimetic preparation of trihydroxybenzacridine pigments from CGA and amino acids such as glycine and lysine, their straightforward purification by gel filtration chromatography, the UHPLC-MS/MS analysis of the purified pigment fraction, and a detailed characterization of the pH-dependent trihydroxybenzacridine chromophore. Similar green pigments were also obtained by analogous reaction of CGA with a low-cost protein, bovine serum albumin, and by simply adding CGA to chicken egg white (CEW) under stirring. Neither the purified pigments from amino acids nor the pigmented CEW exerted significant toxicity against two human cell lines, Caco-2 and HepG2, at doses compatible with common use in food coloring. Additions of the pure pigments or pigmented CEW to different food matrices imparted intense green hues, and the thermal stability of these preparations proved satisfactory up to 90 °C. The potential application of the greening reaction for the sensing of fish deterioration is also disclosed.

Nonphotosynthetic Pigments as Potential Biosignatures

PubMed Central

Cockell, Charles S.; Meadows, Victoria S.

2015-01-01

Abstract Previous work on possible surface reflectance biosignatures for Earth-like planets has typically focused on analogues to spectral features produced by photosynthetic organisms on Earth, such as the vegetation red edge. Although oxygenic photosynthesis, facilitated by pigments evolved to capture photons, is the dominant metabolism on our planet, pigmentation has evolved for multiple purposes to adapt organisms to their environment. We present an interdisciplinary study of the diversity and detectability of nonphotosynthetic pigments as biosignatures, which includes a description of environments that host nonphotosynthetic biologically pigmented surfaces, and a lab-based experimental analysis of the spectral and broadband color diversity of pigmented organisms on Earth. We test the utility of broadband color to distinguish between Earth-like planets with significant coverage of nonphotosynthetic pigments and those with photosynthetic or nonbiological surfaces, using both 1-D and 3-D spectral models. We demonstrate that, given sufficient surface coverage, nonphotosynthetic pigments could significantly impact the disk-averaged spectrum of a planet. However, we find that due to the possible diversity of organisms and environments, and the confounding effects of the atmosphere and clouds, determination of substantial coverage by biologically produced pigments would be difficult with broadband colors alone and would likely require spectrally resolved data. Key Words: Biosignatures—Exoplanets—Halophiles—Pigmentation—Reflectance spectroscopy—Spectral models. Astrobiology 15, 341–361. PMID:25941875

Operation Guiding Light-Scientific Program and Field Plan. The Pilot Field Experiment for NORDA Project Chemical Dynamics in Ocean Frontal Areas

DTIC Science & Technology

1985-03-01

distribution. Samples of suspended partici’lates will also be collected for later image and elemental analysis . 25 Method of analysis for particle...will be flow injection analysis . This method will allow rapid, continuous analysis of seawater nutrients. Measurements will be made at one minute...5 m intervals) as well as from the underway pumping system. Method of pigment analysis for porphyrin and carotenoid pigments will be separation by

Exploring the potential of hyper-spectral imaging for the biogeochemical analysis of varved lake sediments

NASA Astrophysics Data System (ADS)

Butz, Christoph; Grosjean, Martin; Enters, Dirk; Tylmann, Wojciech

2014-05-01

Varved lake sediments have successfully been used to make inferences about past environmental and climate conditions from annual to multi-millennial scales. Among other proxies, concentrations of sedimentary photopigments have been used for temperature reconstructions. However, obtaining well calibrated annually resolved records from sediments still remains challenging. Most laboratory methods used to analyse lake sediments require physical subsampling and are destructive in the process. Hence, temporal resolution and number of data are limited by the amount of material available in the core. Furthermore, for very low sediment accumulation rates annual subsampling is often very difficult or even impossible. To address these problems we explore hyper-spectral imaging as a new method to analyse lake sediments based on their reflectance spectra in the visible and near infrared spectrum. In contrast to other fast and non-destructive methods like X-ray fluorescence, VIS/NIR reflectance spectrometry distinguishes between biogeochemical substances rather than single elements. Rein (2003) has shown that VIS-RS can be used to detect relative concentrations of sedimentary photopigments (e.g. chlorins, carotenoids) and clay minerals. This study presents an advanced approach using a hyper-spectral camera and remote sensing techniques to infer climate proxy data from reflectance spectra of varved lake sediments. Hyper-spectral imaging allows analysing an entire sediment core in a single measurement, producing a spectral dataset with very high spatial (30x30µm/pixel) and spectral resolutions (~1nm) and a higher spectral range (400-1000nm) compared to previously used spectrophotometers. This allows the analysis of data time series at sub-varve scales or spatial mapping of sedimentary substances (e.g. chlorophyll-a and diagenetic products) at very high resolution. The method is demonstrated on varved lake sediments from northern Poland showing the change of the relative concentrations of chlorin pigments within individual varve years. In a next step absolute concentrations of chlorins derived from HPLC measurements have been calibrated to the spectral data using a linear regression model. This results in a very high-resolution dataset of absolute sedimentary pigment concentrations. In a second example µXRF measurements are used to validate a spectral index for clay mineral detection.

Combined SERS and Raman analysis for the identification of red pigments in cross-sections from historic oil paintings.

PubMed

Frano, Kristen A; Mayhew, Hannah E; Svoboda, Shelley A; Wustholz, Kristin L

2014-12-21

The analysis of paint cross-sections can reveal a remarkable amount of information about the layers and materials in a painting without visibly altering the artwork. Although a variety of analytical approaches are used to detect inorganic pigments as well as organic binders, proteins, and lipids in cross-sections, they do not provide for the unambiguous identification of natural, organic colorants. Here, we develop a novel combined surface-enhanced Raman scattering (SERS), light microscopy, and normal Raman scattering (NRS) approach for the identification of red organic and inorganic pigments in paint cross-sections obtained from historic 18th and 19th century oil paintings. In particular, Ag nanoparticles are directly applied to localized areas of paint cross-sections mounted in polyester resin for SERS analysis of the organic pigments. This combined extractionless non-hydrolysis SERS and NRS approach provides for the definitive identification of carmine lake, madder lake, and vermilion in multiple paint layers. To our knowledge, this study represents the first in situ identification of natural, organic pigments within paint cross-sections from oil paintings. Furthermore, the combination of SERS and normal Raman, with light microscopy provides conservators with a more comprehensive understanding of a painting from a single sample and without the need for sample pretreatment.

Color quality of pigments in cochineals (Dactylopius coccus Costa). Geographical origin characterization using multivariate statistical analysis.

PubMed

Méndez, Jesús; González, Mónica; Lobo, M Gloria; Carnero, Aurelio

2004-03-10

The commercial value of a cochineal (Dactylopius coccus Costa) sample is associated with its color quality. Because the cochineal is a legal food colorant, its color quality is generally understood as its pigment content. Simply put, the higher this content, the more valuable the sample is to the market. In an effort to devise a way to measure the color quality of a cochineal, the present study evaluates different parameters of color measurement such as chromatic attributes (L*, and a*), percentage of carminic acid, tint determination, and chromatographic profile of pigments. Tint determination did not achieve this objective because this parameter does not correlate with carminic acid content. On the other hand, carminic acid showed a highly significant correlation (r = - 0.922, p = 0.000) with L* values determined from powdered cochineal samples. The combination of the information from the spectrophotometric determination of carminic acid with that of the pigment profile acquired by liquid chromatography (LC) and the composition of the red and yellow pigment groups, also acquired by LC, enables greater accuracy in judging the quality of the final sample. As a result of this study, it was possible to achieve the separation of cochineal samples according to geographical origin using two statistical techniques: cluster analysis and principal component analysis.

Effect of Monascus purpureus inoculum concentration on pigment production in jackfruit seed flour substrate

NASA Astrophysics Data System (ADS)

Hamdiyati, Yanti; Kusnadi, Yuliani, Lia Amelia

2016-02-01

The used of synthetic dyes have various negative effects on human health. Roomates pigment produced by Monascus purpureus mold can be used as an alternative natural food coloring. The research on the effect of inoculum concentration's M. purpureus to pigment production on the jackfruit seed flour has been done. The objective of research to is to investigate the effect of inoculum concentration's M. purpureus to the production of red, yellow and orange pigment on the jackfruit seed flour. The concentrations used were 0%, 5%, 10%, and 15% (v/w). The result of the data analysed using One-Way ANOVA showed that the inoculum concentration affected the production of red pigment M. purpureus, as well as the data analysis using the Kruskal-Wallis showed that inoculum concentration has influence on the production of yellow and orange pigments. Inoculum concentration of 15% is the optimum concentration for the production of red, yellow and orange pigments with 0:10, 0:50 and 0:20 absorbance units per gram of sample respectively. Based on the results of the research, it can be concluded that inoculum concentration of M. purpureus influenced the production of red, yellow and orange pigments.

The effect of nutrient supply ratios on organic matter dynamics, phytoplankton community composition and diazotrophy in the eastern tropical South Pacific

NASA Astrophysics Data System (ADS)

Meyer, J.; Lavik, G.; Riebesell, U.

2015-12-01

Upwelling of nutrient loaded water masses with low inorganic nitrogen (N) to phosphorus (P) ratios is thought to favor non-Redfield primary production by phytoplankton species adapted to exponential growth. Additionally, an excess of P (P*) in OMZ-influenced waters is also supposed to provide a niche for nitrogen fixing organisms. In order to assess the influence of low inorganic nutrient ratios on the stoichiometry and composition of primary producers, biogeochemical measurements were carried out in the eastern tropical South Pacific during R/V Meteor cruise M93. A succession of different functional types of phytoplankton was observed along onshore - offshore transects with diatoms dominating the productive upwelling region, while haptophytes, cryptophytes and crysophytes prevailed in the more oligotrophic open ocean. Simultaneously, particulate organic nitrogen to phosphorus ratios increased with increasing distance from shore. The stoichiometry of organic matter, however, always exceeded ratios of 16:1, although nutrient supply ratios were below Redfield proportions in the whole sampling area. A considerable amount of P* was detected in the surface ocean layer above the shelf, which decreased as water masses were advected beyond the shelf slope. Phytoplankton pigment analyses with HPLC revealed the existence of diazotrophic marker pigments in the study area, hinting towards a local replenishment of the N-deficit via nitrogen fixation.

Carotenoid accumulation in orange-pigmented Capsicum annuum fruit, regulated at multiple levels

PubMed Central

Rodriguez-Uribe, Laura; Guzman, Ivette; Rajapakse, Wathsala; Richins, Richard D.; O’Connell, Mary A.

2012-01-01

The pericarp of Capsicum fruit is a rich dietary source of carotenoids. Accumulation of these compounds may be controlled, in part, by gene transcription of biosynthetic enzymes. The carotenoid composition in a number of orange-coloured C. annuum cultivars was determined using HPLC and compared with transcript abundances for four carotenogenic enzymes, Psy, LcyB, CrtZ-2, and Ccs determined by qRT-PCR. There were unique carotenoid profiles as well as distinct patterns of transcription of carotenogenic enzymes within the seven orange-coloured cultivars. In one cultivar, ‘Fogo’, carrying the mutant ccs-3 allele, transcripts were detected for this gene, but no CCS protein accumulated. The premature stop termination in ccs-3 prevented expression of the biosynthetic activity to synthesize the capsanthin and capsorubin forms of carotenoids. In two other orange-coloured cultivars, ‘Orange Grande’ and ‘Oriole’, both with wild-type versions of all four carotenogenic enzymes, no transcripts for Ccs were detected and no red pigments accumulated. Finally, in a third case, the orange-coloured cultivar, Canary, transcripts for all four of the wild-type carotenogenic enzymes were readily detected yet no CCS protein appeared to accumulate and no red carotenoids were synthesized. In the past, mutations in Psy and Ccs have been identified as the loci controlling colour in the fruit. Now there is evidence that a non-structural gene may control colour development in Capsicum. PMID:21948863

Carotenoid accumulation in orange-pigmented Capsicum annuum fruit, regulated at multiple levels.

PubMed

Rodriguez-Uribe, Laura; Guzman, Ivette; Rajapakse, Wathsala; Richins, Richard D; O'Connell, Mary A

2012-01-01

The pericarp of Capsicum fruit is a rich dietary source of carotenoids. Accumulation of these compounds may be controlled, in part, by gene transcription of biosynthetic enzymes. The carotenoid composition in a number of orange-coloured C. annuum cultivars was determined using HPLC and compared with transcript abundances for four carotenogenic enzymes, Psy, LcyB, CrtZ-2, and Ccs determined by qRT-PCR. There were unique carotenoid profiles as well as distinct patterns of transcription of carotenogenic enzymes within the seven orange-coloured cultivars. In one cultivar, 'Fogo', carrying the mutant ccs-3 allele, transcripts were detected for this gene, but no CCS protein accumulated. The premature stop termination in ccs-3 prevented expression of the biosynthetic activity to synthesize the capsanthin and capsorubin forms of carotenoids. In two other orange-coloured cultivars, 'Orange Grande' and 'Oriole', both with wild-type versions of all four carotenogenic enzymes, no transcripts for Ccs were detected and no red pigments accumulated. Finally, in a third case, the orange-coloured cultivar, Canary, transcripts for all four of the wild-type carotenogenic enzymes were readily detected yet no CCS protein appeared to accumulate and no red carotenoids were synthesized. In the past, mutations in Psy and Ccs have been identified as the loci controlling colour in the fruit. Now there is evidence that a non-structural gene may control colour development in Capsicum.

Bioactives in Chinese Proprietary Medicine Modulates 5α-Reductase Activity and Gene Expression Associated with Androgenetic Alopecia

PubMed Central

Tan, Justin J. Y.; Pan, Jing; Sun, Lihan; Zhang, Junying; Wu, Chunyong; Kang, Lifeng

2017-01-01

Androgenetic alopecia (AGA) is characterized by a progressive and patterned transformation of thick, pigmented terminal scalp hairs into short, hypo-pigmented vellus-like hairs. The use of Minoxidil and Finasteride to treat AGA are often associated with complications in safety and efficacy. However, herbal remedies are deemed to have lesser side effects in many societies. This study aims to identify potential hair growth properties of individual compounds from a Chinese proprietary medicine known as Yangxue Shengfa capsule (YSC), used in China for many years for improving AGA. Six marker compounds, including 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside (TSG), Chlorogenic acid, Emodin, Ferulic acid, Isoimperatorin, and Paeoniflorin were used for simultaneous HPLC quantification and anti-AGA in-vitro screening. Simultaneous quantification of these components was performed on 75% (v/v) methanol extracts of YSC, using a Welch Ultimate XB-C18 column and gradient elution. Five compounds significantly promoted cell proliferation in cultured immortalized human Dermal Papilla Cells (DPC). Multiple genes associated with the progression of AGA, including IGF-1, DKK-1, and TGF-β1, were found to be regulated by some of these compounds. Interestingly, Ferulic acid and Emodin demonstrated good pharmacological properties against AGA, thereby concluding the potential of these bioactives to be used in the treatment against AGA. PMID:28450835

Online identification of chlorogenic acids, sesquiterpene lactones, and flavonoids in the Brazilian arnica Lychnophora ericoides Mart. (Asteraceae) leaves by HPLC-DAD-MS and HPLC-DAD-MS/MS and a validated HPLC-DAD method for their simultaneous analysis.

PubMed

Gobbo-Neto, Leonardo; Lopes, Norberto P

2008-02-27

Lychnophora ericoides Mart. (Asteraceae, Vernonieae) is a plant, endemic to Brazil, with occurrence restricted to the "cerrado" biome. Traditional medicine employs alcoholic and aqueous-alcoholic preparations of leaves from this species for the treatment of wounds, inflammation, and pain. Furthermore, leaves of L. ericoides are also widely used as flavorings for the Brazilian traditional spirit "cachaça". A method has been developed for the extraction and HPLC-DAD analysis of the secondary metabolites of L. ericoides leaves. This analytical method was validated with 11 secondary metabolites chosen to represent the different classes and polarities of secondary metabolites occurring in L. ericoides leaves, and good responses were obtained for each validation parameter analyzed. The same HPLC analytical method was also employed for online secondary metabolite identification by HPLC-DAD-MS and HPLC-DAD-MS/MS, leading to the identification of di- C-glucosylflavones, coumaroylglucosylflavonols, flavone, flavanones, flavonols, chalcones, goyazensolide, and eremantholide-type sesquiterpene lactones and positional isomeric series of chlorogenic acids possessing caffeic and/or ferulic moieties. Among the 52 chromatographic peaks observed, 36 were fully identified and 8 were attributed to compounds belonging to series of caffeoylferuloylquinic and diferuloylquinic acids that could not be individualized from each other.

Comparison of UPLC and HPLC methods for determination of vitamin C.

PubMed

Klimczak, Inga; Gliszczyńska-Świgło, Anna

2015-05-15

Ultra performance liquid chromatography (UPLC) and high-performance liquid chromatography (HPLC) methods for determination of ascorbic acid (AA) and total AA (TAA) contents (as the sum of AA and dehydroascorbic acid (DHAA) after its reduction to AA) in fruit beverages and in pharmaceutical preparations were compared. Both methods are rapid: total time of analysis was 15 and 6 min for HPLC and UPLC methods, respectively. The methods were validated in terms of linearity, instrument precision, limits of detection (LOD) and quantification (LOQ), accuracy and recovery. Intra- and inter-day instrument precisions for fruit juices, expressed as RSD, were 2.2% and 2.4% for HPLC, respectively, and 1.7% and 1.9% for UPLC, respectively. For vitamin C tablets, inter- and intra-day precisions were 0.4% and 0.5%, respectively (HPLC), and 0.5% and 0.3%, respectively (UPLC). Both methods were sensitive: LOD was 0.049 μg/mL for HPLC and 0.024 μg/mL for UPLC while LOQs were 0.149 and 0.073 μg/mL for HPLC and UPLC, respectively. These methods could be useful in the routine qualitative and quantitative analysis of AA or TAA in pharmaceutical preparations or fruit beverages. However, UPLC method is more sensitive, faster and consumes less eluent. Copyright © 2014 Elsevier Ltd. All rights reserved.

A novel approach for quantitation of nonderivatized sialic acid in protein therapeutics using hydrophilic interaction chromatographic separation and nano quantity analyte detection.

PubMed

Chemmalil, Letha; Suravajjala, Sreekanth; See, Kate; Jordan, Eric; Furtado, Marsha; Sun, Chong; Hosselet, Stephen

2015-01-01

This paper describes a novel approach for the quantitation of nonderivatized sialic acid in glycoproteins, separated by hydrophilic interaction chromatography, and detection by Nano Quantity Analyte Detector (NQAD). The detection technique of NQAD is based on measuring change in the size of dry aerosol and converting the particle count rate into chromatographic output signal. NQAD detector is suitable for the detection of sialic acid, which lacks sufficiently active chromophore or fluorophore. The water condensation particle counting technology allows the analyte to be enlarged using water vapor to provide highest sensitivity. Derivatization-free analysis of glycoproteins using HPLC/NQAD method with PolyGLYCOPLEX™ amide column is well correlated with HPLC method with precolumn derivatization using 1, 2-diamino-4, 5-methylenedioxybenzene (DMB) as well as the Dionex-based high-pH anion-exchange chromatography (or ion chromatography) with pulsed amperometric detection (HPAEC-PAD). With the elimination of derivatization step, HPLC/NQAD method is more efficient than HPLC/DMB method. HPLC/NQAD method is more reproducible than HPAEC-PAD method as HPAEC-PAD method suffers high variability because of electrode fouling during analysis. Overall, HPLC/NQAD method offers broad linear dynamic range as well as excellent precision, accuracy, repeatability, reliability, and ease of use, with acceptable comparability to the commonly used HPAEC-PAD and HPLC/DMB methods. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

CAPN3, DCT, MLANA and TYRP1 are overexpressed in skin of vitiligo vulgaris Mexican patients.

PubMed

Salinas-Santander, Mauricio; Trevino, Víctor; De la Rosa-Moreno, Eduardo; Verduzco-Garza, Bárbara; Sánchez-Domínguez, Celia N; Cantú-Salinas, Cristina; Ocampo-Garza, Jorge; Lagos-Rodríguez, Armando; Ocampo-Candiani, Jorge; Ortiz-López, Rocio

2018-03-01

Vitiligo is a disorder causing skin depigmentation, in which several factors have been proposed for its pathogenesis: Environmental, genetic and biological aspects of melanocytes, even those of the surrounding keratinocytes. However, the lack of understanding of the mechanisms has complicated the task of predicting the development and progression. The present study used microarray analysis to characterize the transcriptional profile of skin from Vitiligo Vulgaris (VV) patients and the identified transcripts were validated using targeted high-throughput RNA sequencing in a broader set of patients. For microarrays, mRNA was taken from 20 skin biopsies of 10 patients with VV (pigmented and depigmented skin biopsy of each), and 5 biopsies of healthy subjects matched for age and sex were used as a control. A signature was identified that contains the expression pattern of 722 genes between depigmented vitiligo skin vs. healthy control, 1,108 between the pigmented skin of vitiligo vs. healthy controls and 1,927 between pigmented skin, depigmented vitiligo and healthy controls (P<0.05; false discovery rate, <0.1). When comparing the pigmented and depigmented skin of patients with vitiligo, which reflects the real difference between both skin types, 5 differentially expressed genes were identified and further validated in 45 additional VV patients by RNA sequencing. This analysis showed significantly higher RNA levels of calpain-3, dopachrome tautomerase, melan-A and tyrosinase-related protein-1 genes. The data revealed that the pigmented skin of vitiligo is already affected at the level of gene expression and that the main differences between pigmented and non-pigmented skin are explained by the expression of genes associated with pigment metabolism.

The influence of visible light and inorganic pigments on fluorescence excitation emission spectra of egg-, casein- and collagen-based painting media

NASA Astrophysics Data System (ADS)

Nevin, A.; Anglos, D.; Cather, S.; Burnstock, A.

2008-07-01

Spectrofluorimetric analysis of proteinaceous binding media is particularly promising because proteins employed in paintings are often fluorescent and media from different sources have significantly different fluorescence spectral profiles. Protein-based binding media derived from eggs, milk and animal tissue have been used for painting and for conservation, but their analysis using non-destructive techniques is complicated by interferences with pigments, their degradation and their low concentration. Changes in the fluorescence excitation emission spectra of films of binding media following artificial ageing to an equivalent of 50 and 100 years of museum lighting include the reduction of bands ascribed to tyrosine, tryptophan and Maillard reaction products and an increase in fluorescent photodegradation. Fluorescence of naturally aged paint is dependent on the nature of the pigment present and, with egg-based media, in comparison with un-pigmented films, emissions ascribed to amino acids are more pronounced.

The comparative study of four Portuguese sixteenth-century illuminated Manueline Charters based on spectroscopy and chemometrics analysis

NASA Astrophysics Data System (ADS)

Miguel, Catarina; Barrocas-Dias, Cristina; Ferreira, Teresa; Candeias, António

2017-01-01

The comparative study based on spectroscopic analysis of the materials used to produce four sixteenth-century Manueline Charters (the Charters of Alcochete, Terena, Alandroal and Évora) was performed following a systematic analytical approach. SEM-EDS, μ-Raman and μ-FTIR analysis highlighted interesting features between them, namely the use of different pigments and colourants (such as different green and yellow pigments), the presence of pigments alterations and the use of a non-expected extemporaneous material (with the presence of titanium white in the Charter of Alcochete). Principal component analysis restricted to the C-H absorption region (3000-2840 cm-1) was applied to 36 infrared spectra of blue historical samples from the Charters of Alcochete, Terena, Alandroal and Évora, suggesting the use of a mixture of a triglyceride and polysaccharide as binder.

Microanalysis study of archaeological mural samples containing Maya blue pigment

NASA Astrophysics Data System (ADS)

Sánchez del Río, M.; Martinetto, P.; Somogyi, A.; Reyes-Valerio, C.; Dooryhée, E.; Peltier, N.; Alianelli, L.; Moignard, B.; Pichon, L.; Calligaro, T.; Dran, J.-C.

2004-10-01

Elemental analysis by X-ray fluorescence and particle induced X-ray emission is applied to the study of several Mesoamerican mural samples containing blue pigments. The most characteristic blue pigment is Maya blue, a very stable organo-clay complex original from Maya culture and widely used in murals, pottery and sculptures in a vast region of Mesoamerica during the pre-hispanic time (from VIII century) and during the colonization until 1580. The mural samples come from six different archaeological sites (four pre-hispanic and two from XVI century colonial convents). The correlation between the presence of some elements and the pigment colour is discussed. From the comparative study of the elemental concentration, some conclusions are drawn on the nature of the pigments and the technology used.

Silk fibroin film from golden-yellow Bombyx mori is a biocomposite that contains lutein and promotes axonal growth of primary neurons.

PubMed

Pistone, Assunta; Sagnella, Anna; Chieco, Camilla; Bertazza, Gianpaolo; Varchi, Greta; Formaggio, Francesco; Posati, Tamara; Saracino, Emanuela; Caprini, Marco; Bonetti, Simone; Toffanin, Stefano; Di Virgilio, Nicola; Muccini, Michele; Rossi, Federica; Ruani, Giampiero; Zamboni, Roberto; Benfenati, Valentina

2016-05-01

The use of doped silk fibroin (SF) films and substrates from Bombyx mori cocoons for green nanotechnology and biomedical applications has been recently highlighted. Cocoons from coloured strains of B. mori, such as Golden-Yellow, contain high levels of pigments that could have a huge potential for the fabrication of SF based biomaterials targeted to photonics, optoelectronics and neuroregenerative medicine. However, the features of extracted and regenerated SF from cocoons of B. mori Golden-Yellow strain have never been reported. Here we provide a chemophysical characterization of regenerated silk fibroin (RSF) fibers, solution, and films obtained from cocoons of a Golden-Yellow strain of B. mori, by SEM, (1) H-NMR, HPLC, FT-IR, Raman and UV-Vis spectroscopy. We found that the extracted solution and films from B. mori Golden-Yellow fibroin displayed typical Raman spectroscopic and optical features of carotenoids. HPLC-analyses revealed that lutein was the carotenoid contained in the fiber and RSF biopolymer from yellow cocoons. Notably, primary neurons cultured on yellow SF displayed a threefold higher neurite length than those grown of white SF films. The results we report pave the way to expand the potential use of yellow SF in the field of neuroregenerative medicine and provide green chemistry approaches in biomedicine. © 2016 Wiley Periodicals, Inc.

Untargeted Metabolomics Approach in Halophiles: Understanding the Biodeterioration Process of Building Materials

PubMed Central

Adamiak, Justyna; Bonifay, Vincent; Otlewska, Anna; Sunner, Jan A.; Beech, Iwona B.; Stryszewska, Teresa; Kańka, Stanisław; Oracz, Joanna; Żyżelewicz, Dorota; Gutarowska, Beata

2017-01-01

The aim of the study was to explore the halophile metabolome in building materials using untargeted metabolomics which allows for broad metabolome coverage. For this reason, we used high-performance liquid chromatography interfaced to high-resolution mass spectrometry (HPLC/HRMS). As an alternative to standard microscopy techniques, we introduced pioneering Coherent Anti-stokes Raman Scattering Microscopy (CARS) to non-invasively visualize microbial cells. Brick samples saturated with salt solution (KCl, NaCl (two salinity levels), MgSO4, Mg(NO3)2), were inoculated with the mixture of preselected halophilic microorganisms, i.e., bacteria: Halobacillus styriensis, Halobacillus naozhouensis, Halobacillus hunanensis, Staphylococcus succinus, Marinococcus halophilus, Virgibacillus halodenitryficans, and yeast: Sterigmatomyces halophilus and stored at 28°C and 80% relative humidity for a year. Metabolites were extracted directly from the brick samples and measured via HPLC/HRMS in both positive and negative ion modes. Overall, untargeted metabolomics allowed for discovering the interactions of halophilic microorganisms with buildings materials which together with CARS microscopy enabled us to elucidate the biodeterioration process caused by halophiles. We observed that halophile metabolome was differently affected by different salt solutions. Furthermore, we found indications for haloadaptive strategies and degradation of brick samples due to microbial pigment production as a salt stress response. Finally, we detected changes in lipid content related to changes in the structure of phospholipid bilayers and membrane fluidity. PMID:29321766

HPLC ANALYSIS OF VINCLOZOLIN AND ITS METABOLITES IN SERUM

EPA Science Inventory

HPLC ANALYSIS OF VINCLOZOLIN AND ITS METABOLITES IN SERUM. A Sierra-Santoyo1,2, H A Barton1 and M F Hughes1. 1US EPA, ORD, NHEERL, ETD, RTP, NC; 2Toxicology Section, CINVESTAV-IPN, Mexico City, Mexico.

The fungicide vinclozolin (V) is used predominantly for treatment...

Industrial application of green chromatography--I. Separation and analysis of niacinamide in skincare creams using pure water as the mobile phase.

PubMed

Yang, Yu; Strickland, Zackary; Kapalavavi, Brahmam; Marple, Ronita; Gamsky, Chris

2011-03-15

In this work, chromatographic separation of niacin and niacinamide using pure water as the sole component in the mobile phase has been investigated. The separation and analysis of niacinamide have been optimized using three columns at different temperatures and various flow rates. Our results clearly demonstrate that separation and analysis of niacinamide from skincare products can be achieved using pure water as the eluent at 60°C on a Waters XTerra MS C18 column, a Waters XBridge C18 column, or at 80°C on a Hamilton PRP-1 column. The separation efficiency, quantification quality, and analysis time of this new method are at least comparable with those of the traditional HPLC methods. Compared with traditional HPLC, the major advantage of this newly developed green chromatography technique is the elimination of organic solvents required in the HPLC mobile phase. In addition, the pure water chromatography separations described in this work can be directly applied in industrial plant settings without further modification of the existing HPLC equipment. Copyright © 2011 Elsevier B.V. All rights reserved.

Synthesis and characterization of hematite pigment obtained from a steel waste industry.

PubMed

Prim, S R; Folgueras, M V; de Lima, M A; Hotza, D

2011-09-15

Pigments that meet environmental and technology requirements are the focus of the research in the ceramic sector. This study focuses on the synthesis of ceramic pigment by encapsulation of hematite in crystalline and amorphous silica matrix. Iron oxide from a metal sheet rolling process was used as chromophore. A different content of hematite and silica was homogenized by conventional and high energy milling. The powders obtained after calcinations between 1050 and 1200 °C for 2h were characterized by X-ray diffraction and SEM analysis. The pigments were applied to ceramic enamel and porcelain body. The effect of pigment was measured by comparing L*a*b* values of the heated samples. Results showed that the color developed is influenced by variables such as oxide content employed, conditions of milling and processing temperature. The results showed that the use of pigment developed does not interfere in microstructural characteristics of pigmented material. The best hue was obtained from samples with 15 wt% of chromophore, heated at 1200 °C in amorphous silica matrix. Copyright © 2011 Elsevier B.V. All rights reserved.

Composition of prehistoric rock-painting pigments from Egypt (Gilf Kébir area).

PubMed

Darchuk, L; Rotondo, G Gatto; Swaenen, M; Worobiec, A; Tsybrii, Z; Makarovska, Y; Van Grieken, R

2011-12-01

The composition of rock-painting pigments from Egypt (Gilf Kebia area) has been analyzed by means of molecular spectroscopy such as Fourier transform infrared and micro-Raman spectroscopy and scanning electron microscopy coupled to an energy dispersive X-ray spectrometer and X-ray fluorescence analysis. Red and yellow pigments were recognized as red and yellow ochre with additional rutile. Copyright © 2011 Elsevier B.V. All rights reserved.

Analysis of formononetin from black cohosh (Actaea racemosa).

PubMed

Jiang, B; Kronenberg, F; Balick, M J; Kennelly, E J

2006-07-01

Black cohosh has been widely used as an herbal medicine for the treatment of symptoms related to menopause in America and Europe during the past several decades, but the bioactive constituents are still unknown. Formononetin is an isoflavone with known estrogen-like activity. This compound was first reported to be isolated from black cohosh in 1985, but subsequent research in 2002 using HPLC-PDA and LC-MS revealed no evidence to show the presence of formononetin in 13 populations of American black cohosh. A more recent report published in 2004 claimed to detect formononetin in an extract of black cohosh rhizomes using a TLC-fluorescent densitometry method. To further resolve these conflicting reports, we analyzed black cohosh roots and rhizomes for the presence of formononetin, using a combined TLC, HPLC-PDA and LC-MS method. We examined both methanolic and aqueous methanolic black cohosh extracts by HPLC-PDA and LC-MS methods, and did not detect formononetin in any extracts. We further determined the limits of detection of formononetin by HPLC-PDA and LC-MS. Our experimental results indicated that the sensitivity and accuracy of the HPLC-PDA and LC-MS methods for the analysis of formononetin were slightly higher than those of the reported fluorescent method, suggesting that the HPLC-PDA and LC-MS methods were reliable for the analysis of formononetin from black cohosh. We also repeated the reported TLC method to concentrate two fractions from a modern black cohosh sample and an 86-year-old black cohosh sample, respectively, and then analyzed these two fractions for formononetin using the HPLC-PDA and LC-MS method instead of the fluorescent method. Formononetin was not detected by HPLC-PDA or LC-MS. From the results of the present study it is not reasonable to attribute the estrogen-like activity of black cohosh extracts to formononetin.

Ultrastructural analysis of the pigment dispersion syndrome in DBA/2J mice.

PubMed

Schraermeyer, Mareike; Schnichels, Sven; Julien, Sylvie; Heiduschka, Peter; Bartz-Schmidt, Karl-Ulrich; Schraermeyer, Ulrich

2009-11-01

To characterise ocular pigment abnormalities associated with iris atrophy in DBA/2J mice as a model for human pigment dispersion syndrome. Immunohistochemistry, electron and light microscopy were performed to examine the eyes of DBA/2J mice ranging in age from 2.5 to 18 months old. The focus of our study was the description of the ultrastructural modifications in the irides of DBA/2J mice. The DBA/2J mice presented modifications in the melanosomes in all the pigmented parts of the eye, including the retinal pigment epithelial cells and choroidal melanocytes of the ciliary pigment epithelium. The extracellular matrix of the iris stroma disappeared with ageing. Pigmented cells detached from the iris and migrated into the trabecular meshwork exclusively on the anterior iris surface. These cells were identified as macrophages by immunohistochemistry and electron microscopy. There was no evidence that melanocytes or iris pigment epithelial cells migrated into the trabecular meshwork, but they became more and more depigmented. The aqueous outflow was blocked by pigment-laden cells, but not by cellular debris or melanosomes. No substantial amount of extracellular melanosomes was observed. The morphology of melanosomes is aberrant in all pigment cells in the eyes of DBA/2J mice. We conclude that the disease process begins with the transfer of both immature melanosomes from the iris pigment epithelium (IPE) and melanocytes to macrophages, which subsequently migrate into the trabecular meshwork. Accumulating macrophages cause a blockade of the chamber angle. As the disease progresses, the IPE, melanocytes and iris stroma, including blood vessels, disappear, leading to iris atrophy. It is speculated that the loss of these pigment cells is partly caused by reduction of the iris stroma.

Analysis of carbohydrates in Fusarium verticillioides using size-exclusion HPLC – DRI and direct analysis in real time ionization – time-of-flight – mass spectrometry (DART-MS)

USDA-ARS?s Scientific Manuscript database

Direct analysis in real time ionization – time-of-flight – mass spectrometry (DART-MS) and size-exclusion HPLC – DRI are used, respectively, to qualitatively and quantitatively determine the carbohydrates extracted from the corn rot fungus Fusarium verticillioides. In situ permethylation in the DART...

Comparison between non-invasive methods used on paintings by Goya and his contemporaries: hyperspectral imaging vs. point-by-point spectroscopic analysis.

PubMed

Daniel, Floréal; Mounier, Aurélie; Pérez-Arantegui, Josefina; Pardos, Carlos; Prieto-Taboada, Nagore; Fdez-Ortiz de Vallejuelo, Silvia; Castro, Kepa

2017-06-01

The development of non-invasive techniques for the characterization of pigments is crucial in order to preserve the integrity of the artwork. In this sense, the usefulness of hyperspectral imaging was demonstrated. It allows pigment characterization of the whole painting. However, it also sometimes requires the complementation of other point-by-point techniques. In the present article, the advantages of hyperspectral imaging over point-by-point spectroscopic analysis were evaluated. For that purpose, three paintings were analysed by hyperspectral imaging, handheld X-ray fluorescence and handheld Raman spectroscopy in order to determine the best non-invasive technique for pigment identifications. Thanks to this work, the main pigments used in Aragonese artworks, and especially in Goya's paintings, were identified and mapped by imaging reflection spectroscopy. All the analysed pigments corresponded to those used at the time of Goya. Regarding the techniques used, the information obtained by the hyperspectral imaging and point-by-point analysis has been, in general, different and complementary. Given this fact, selecting only one technique is not recommended, and the present work demonstrates the usefulness of the combination of all the techniques used as the best non-invasive methodology for the pigments' characterization. Moreover, the proposed methodology is a relatively quick procedure that allows a larger number of Goya's paintings in the museum to be surveyed, increasing the possibility of obtaining significant results and providing a chance for extensive comparisons, which are relevant from the point of view of art history issues.

Multiple fingerprinting analyses in quality control of Cassiae Semen polysaccharides.

PubMed

Cheng, Jing; He, Siyu; Wan, Qiang; Jing, Pu

2018-03-01

Quality control issue overshadows potential health benefits of Cassiae Semen due to the analytic limitations. In this study, multiple-fingerprint analysis integrated with several chemometrics was performed to assess the polysaccharide quality of Cassiae Semen harvested from different locations. FT-IR, HPLC, and GC fingerprints of polysaccharide extracts from the authentic source were established as standard profiles, applying to assess the quality of foreign sources. Analyses of FT-IR fingerprints of polysaccharide extracts using either Pearson correlation analysis or principal component analysis (PCA), or HPLC fingerprints of partially hydrolyzed polysaccharides with PCA, distinguished the foreign sources from the authentic source. However, HPLC or GC fingerprints of completely hydrolyzed polysaccharides couldn't identify all foreign sources and the methodology using GC is quite limited in determining the monosaccharide composition. This indicates that FT-IR/HPLC fingerprints of non/partially-hydrolyzed polysaccharides, respectively, accompanied by multiple chemometrics methods, might be potentially applied in detecting and differentiating sources of Cassiae Semen. Copyright © 2018 Elsevier B.V. All rights reserved.

Development and Validation of an HPLC Method for Karanjin in Pongamia pinnata linn. Leaves.

PubMed

Katekhaye, S; Kale, M S; Laddha, K S

2012-01-01

A rapid, simple and specific reversed-phase HPLC method has been developed for analysis of karanjin in Pongamia pinnata Linn. leaves. HPLC analysis was performed on a C(18) column using an 85:13.5:1.5 (v/v) mixtures of methanol, water and acetic acid as isocratic mobile phase at a flow rate of 1 ml/min. UV detection was at 300 nm. The method was validated for accuracy, precision, linearity, specificity. Validation revealed the method is specific, accurate, precise, reliable and reproducible. Good linear correlation coefficients (r(2)>0.997) were obtained for calibration plots in the ranges tested. Limit of detection was 4.35 μg and limit of quantification was 16.56 μg. Intra and inter-day RSD of retention times and peak areas was less than 1.24% and recovery was between 95.05 and 101.05%. The established HPLC method is appropriate enabling efficient quantitative analysis of karanjin in Pongamia pinnata leaves.

Development and Validation of an HPLC Method for Karanjin in Pongamia pinnata linn. Leaves

PubMed Central

Katekhaye, S; Kale, M. S.; Laddha, K. S.

2012-01-01

A rapid, simple and specific reversed-phase HPLC method has been developed for analysis of karanjin in Pongamia pinnata Linn. leaves. HPLC analysis was performed on a C18 column using an 85:13.5:1.5 (v/v) mixtures of methanol, water and acetic acid as isocratic mobile phase at a flow rate of 1 ml/min. UV detection was at 300 nm. The method was validated for accuracy, precision, linearity, specificity. Validation revealed the method is specific, accurate, precise, reliable and reproducible. Good linear correlation coefficients (r2>0.997) were obtained for calibration plots in the ranges tested. Limit of detection was 4.35 μg and limit of quantification was 16.56 μg. Intra and inter-day RSD of retention times and peak areas was less than 1.24% and recovery was between 95.05 and 101.05%. The established HPLC method is appropriate enabling efficient quantitative analysis of karanjin in Pongamia pinnata leaves. PMID:23204626

Direct injection analysis of fatty and resin acids in papermaking process waters by HPLC/MS.

PubMed

Valto, Piia; Knuutinen, Juha; Alén, Raimo

2011-04-01

A novel HPLC-atmospheric pressure chemical ionization/MS (HPLC-APCI/MS) method was developed for the rapid analysis of selected fatty and resin acids typically present in papermaking process waters. A mixture of palmitic, stearic, oleic, linolenic, and dehydroabietic acids was separated by a commercial HPLC column (a modified stationary C(18) phase) using gradient elution with methanol/0.15% formic acid (pH 2.5) as a mobile phase. The internal standard (myristic acid) method was used to calculate the correlation coefficients and in the quantitation of the results. In the thorough quality parameters measurement, a mixture of these model acids in aqueous media as well as in six different paper machine process waters was quantitatively determined. The measured quality parameters, such as selectivity, linearity, precision, and accuracy, clearly indicated that, compared with traditional gas chromatographic techniques, the simple method developed provided a faster chromatographic analysis with almost real-time monitoring of these acids. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Authentication and Quantitation of Fraud in Extra Virgin Olive Oils Based on HPLC-UV Fingerprinting and Multivariate Calibration

PubMed Central

Carranco, Núria; Farrés-Cebrián, Mireia; Saurina, Javier

2018-01-01

High performance liquid chromatography method with ultra-violet detection (HPLC-UV) fingerprinting was applied for the analysis and characterization of olive oils, and was performed using a Zorbax Eclipse XDB-C8 reversed-phase column under gradient elution, employing 0.1% formic acid aqueous solution and methanol as mobile phase. More than 130 edible oils, including monovarietal extra-virgin olive oils (EVOOs) and other vegetable oils, were analyzed. Principal component analysis results showed a noticeable discrimination between olive oils and other vegetable oils using raw HPLC-UV chromatographic profiles as data descriptors. However, selected HPLC-UV chromatographic time-window segments were necessary to achieve discrimination among monovarietal EVOOs. Partial least square (PLS) regression was employed to tackle olive oil authentication of Arbequina EVOO adulterated with Picual EVOO, a refined olive oil, and sunflower oil. Highly satisfactory results were obtained after PLS analysis, with overall errors in the quantitation of adulteration in the Arbequina EVOO (minimum 2.5% adulterant) below 2.9%. PMID:29561820

[Preparation of flavonoid reference standards from Scutellariae Radix under the guidance of high performance liquid chromatography-mass spectrometry analysis].

PubMed

Guo, Henan; Yang, Xuedong; Liu, Jun; Zheng, Wenfeng

2012-07-01

Flavonoid reference standards were targeted-prepared from Scutellariae Radix under the guidance of high performance liquid chromatography-mass spectrometry (HPLC-MS) analysis. With HPLC-MS analysis of Scutellariae Radix, 19 flavonoid components were identified by analyzing and comparing their retention times, ultraviolet spectra, and mass spectrometry data with literature. The separation and purification protocols of all targeted flavonoid reference standards were optimally designed according to the results of HPLC-MS analysis and related literature. The ethanol extract of Scutellariae Radix was suspended in water and extracted with petroleum ether, ethyl acetate, and n-butanol successively. The ethyl acetate extract and n-butanol extract were separately subjected to primary separation by low pressure reverse phase preparative chromatography. Then the fractions containing targeted compounds were further purified by low pressure reverse and normal phases preparative chromatography. Finally, baicalin and wogonoside reference standards were obtained from n-butanol extract; baicaelin, wogonin, and oroxylin A reference standards were obtained from ethyl acetate extract. The structures of the 5 reference standards were identified by mass spectrometry (MS) and 1H nuclear magnetic resonance (1H NMR) spectroscopy. The HPLC analytical results showed that the purities of the 5 reference standards were all above 98%. It is demonstrated that the rapid targeted-preparation method under the guidance of the HPLC-MS analysis is applicable for the isolation and preparation of chemical components in traditional Chinese medicines.

Simultaneous HPLC quantitative analysis of active compounds in leaves of Moringa oleifera Lam.

PubMed

Vongsak, Boonyadist; Sithisarn, Pongtip; Gritsanapan, Wandee

2014-08-01

Moringa oleifera Lam. has been used as a traditional medicine for the treatment of numerous diseases. A simultaneous high-performance liquid chromatography (HPLC) analysis was developed and validated for the determination of the contents of crypto-chlorogenic acid, isoquercetin and astragalin, the primary antioxidative compounds, in M. oleifera leaves. HPLC analysis was successfully conducted by using a Hypersil BDS C18 column, eluted with a gradient of methanol-1% acetic acid with a flow rate of 1 mL/min, and detected at 334 nm. Parameters for the validation included linearity, precision, accuracy and limits of detection and quantitation. The developed HPLC method was precise, with relative standard deviation < 2%. The recovery values of crypto-chlorogenic acid, isoquercetin and astragalin in M. oleifera leaf extracts were 98.50, 98.47 and 98.59%, respectively. The average contents of these compounds in the dried ethanolic extracts of the leaves of M. oleifera collected from different regions of Thailand were 0.081, 0.120 and 0.153% (w/w), respectively. The developed HPLC method was appropriate and practical for the simultaneous analysis of crypto-chlorogenic acid, isoquercetin and astragalin in the leaf extract of M. oleifera. This work is valuable as guidance for the standardization of the leaf extracts and pharmaceutical products of M. oleifera. © The Author [2013]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Pyridylamination as a means of analyzing complex sugar chains

PubMed Central

Hase, Sumihiro

2010-01-01

Herein, I describe pyridylamination for versatile analysis of sugar chains. The reducing ends of the sugar chains are tagged with 2-aminopyridine and the resultant chemically stable fluorescent derivatives are used for structural/functional analysis. Pyridylamination is an effective “operating system” for increasing sensitivity and simplifying the analytical procedures including mass spectrometry and NMR. Excellent separation of isomers is achieved by reversed-phase HPLC. However, separation is further improved by two-dimensional HPLC, which involves a combination of reversed-phase HPLC and size-fractionation HPLC. Moreover, a two-dimensional HPLC map is also useful for structural analysis. I describe a simple procedure for preparing homogeneous pyridylamino sugar chains that is less laborious than existing techniques and can be used for functional analysis (e.g., sugar-protein interaction). This novel approach was applied and some of the results are described: i) a glucosyl-serine type sugar chain found in blood coagulation factors; ii) discovery of endo-β-mannosidase (EC 3.2.1.152) and a new type plant α1,2-l-fucosidase; and iii) novel substrate specificity of a cytosolic α-mannosidase. Moreover, using homogeneous sugar chains of a size similar to in vivo substrates we were able to analyze interactions between sugar chains and proteins such as enzymes and lectins in detail. Interestingly, our studies reveal that some enzymes recognize a wider region of the substrate than anticipated. PMID:20431262

Crystalline Organic Pigment-Based Field-Effect Transistors.

PubMed

Zhang, Haichang; Deng, Ruonan; Wang, Jing; Li, Xiang; Chen, Yu-Ming; Liu, Kewei; Taubert, Clinton J; Cheng, Stephen Z D; Zhu, Yu

2017-07-05

Three conjugated pigment molecules with fused hydrogen bonds, 3,7-diphenylpyrrolo[2,3-f]indole-2,6(1H,5H)-dione (BDP), (E)-6,6'-dibromo-[3,3'-biindolinylidene]-2,2'-dione (IIDG), and 3,6-di(thiophen-2-yl)-2,5-dihydropyrrolo-[3,4-c]pyrrole-1,4-dione (TDPP), were studied in this work. The insoluble pigment molecules were functionalized with tert-butoxylcarbonyl (t-Boc) groups to form soluble pigment precursors (BDP-Boc, IIDG-Boc, and TDPP-Boc) with latent hydrogen bonding. The single crystals of soluble pigment precursors were obtained. Upon simple thermal annealing, the t-Boc groups were removed and the soluble pigment precursor molecules with latent hydrogen bonding were converted into the original pigment molecules with fused hydrogen bonding. Structural analysis indicated that the highly crystalline soluble precursors were directly converted into highly crystalline insoluble pigments, which are usually only achievable by gas-phase routes like physical vapor transport. The distinct crystal structure after the thermal annealing treatment suggests that fused hydrogen bonding is pivotal for the rearrangement of molecules to form a new crystal in solid state, which leads to over 2 orders of magnitude enhancement in charge mobility in organic field-effect transistor (OFET) devices. This work demonstrated that crystalline OFET devices with insoluble pigment molecules can be fabricated by their soluble precursors. The results indicated that a variety of commercially available conjugated pigments could be potential active materials for high-performance OFETs.

Pigment identification in artwork using graphite furnace atomic absorption spectrometry.

PubMed

Goltz, D M; Coombs, J; Marion, C; Cloutis, E; Gibson, J; Attas, M; Choo-Smith, L-P; Collins, C

2004-06-17

The use of a sampling technique is described for the identification of metals from inorganic pigments in paint. The sampling technique involves gently contacting a cotton swab with the painted surface to physically remove a minute quantity ( approximately 1-2mug) of pigment. The amount of material removed from the painted surface is invisible to the unaided eye and does not cause any visible effect to the painted surface. The cotton swab was then placed in a 1.5ml polystyrene beaker containing HNO(3) to extract pigment metals prior to analysis using graphite furnace atomic absorption spectrometry (GFAAS). GFAAS is well suited for identifying pigment metals since it requires small samples and many pigments consist of main group elements (e.g. Al) as well as transition metals (e.g. Zn, Fe and Cd). Using Cd (cadmium red) as the test element, the reproducibility of sampling a paint surface with the cotton swab was approximately 13% in either a water or oil medium. To test the feasibility of cotton sampling for pigment identification, samples were obtained from paintings (watercolour and oil) of a local collection. Raman spectra provided complementary information to the GFAAS, which together are essential for positive identification of some pigments. For example, GFAAS indicated the presence of Cu, but the Raman spectra positively identified the modern copper pigment phthalocyanine green (Cu(C(32)Cl(16)N(8)). Both Raman spectroscopy and GFAAS were useful for identifying ZnO as a white pigment.

Improved method for HPLC analysis of polyamines, agmatine and aromatic monoamines in plant tissue

NASA Technical Reports Server (NTRS)

Slocum, R. D.; Flores, H. E.; Galston, A. W.; Weinstein, L. H.

1989-01-01

The high performance liquid chromatographic (HPLC) method of Flores and Galston (1982 Plant Physiol 69: 701) for the separation and quantitation of benzoylated polyamines in plant tissues has been widely adopted by other workers. However, due to previously unrecognized problems associated with the derivatization of agmatine, this important intermediate in plant polyamine metabolism cannot be quantitated using this method. Also, two polyamines, putrescine and diaminopropane, also are not well resolved using this method. A simple modification of the original HPLC procedure greatly improves the separation and quantitation of these amines, and further allows the simulation analysis of phenethylamine and tyramine, which are major monoamine constituents of tobacco and other plant tissues. We have used this modified HPLC method to characterize amine titers in suspension cultured carrot (Daucas carota L.) cells and tobacco (Nicotiana tabacum L.) leaf tissues.

Immunoassay screening of lysergic acid diethylamide (LSD) and its confirmation by HPLC and fluorescence detection following LSD ImmunElute extraction.

PubMed

Grobosch, T; Lemm-Ahlers, U

2002-04-01

In all, 3872 urine specimens were screened for lysergic acid diethylamide (LSD) using the CEDIA DAU LSD assay. Forty-eight samples, mainly from psychiatric patients or drug abusers, were found to be LSD positive, but only 13 (27%) of these could be confirmed by high-performance liquid chromatography with fluorescence detection (HPLC-FLD) following immunoaffinity extraction (IAE). Additional analysis for LSD using the DPC Coat-a-Count RIA was performed to compare the two immunoassay screening methods. Complete agreement between the DPC RIA assay and HPLC-FLD results was observed at concentrations below a cutoff concentration of 500 pg/mL. Samples that were LSD positive in the CEDIA DAU assay but not confirmed by HPLC-FLD were also investigated for interfering compounds using REMEDI HS drug-profiling system. REMEDI HS analysis identified 15 compounds (parent drugs and metabolites) that are believed to cross-react in the CEDIA DAU LSD assay: ambroxol, prilocaine, pipamperone, diphenhydramine, metoclopramide, amitriptyline, doxepine, atracurium, bupivacaine, doxylamine, lidocaine, mepivacaine, promethazine, ranitidine, and tramadole. The IAE/HPLC-FLD combination is rapid, easy to perform and reliable. It can reduce costs when standard, rather than more advanced, HPLC equipment is used, especially for labs that perform analyses for LSD infrequently. The chromatographic analysis of LSD, nor-LSD, and iso-LSD is not influenced by any of the tested cross-reacting compounds even at a concentration of 100 ng/mL.

Comparison of UHPLC and HPLC in benzodiazepines analysis of postmortem samples: a case-control study.

PubMed

Behnoush, Behnam; Sheikhazadi, Ardeshir; Bazmi, Elham; Fattahi, Akbar; Sheikhazadi, Elham; Saberi Anary, Seyed Hossein

2015-04-01

The aim of this study was to compare system efficiency and analysis duration regarding the solvent consumption and system maintenance in high-pressure liquid chromatography (HPLC) and ultra high-pressure liquid chromatography (UHPLC). In a case-control study, standard solutions of 7 benzodiazepines (BZs) and 73 biological samples such as urine, tissue, stomach content, and bile that screened positive for BZs were analyzed by HPLC and UHPLC in laboratory of forensic toxicology during 2012 to 2013. HPLC analysis was performed using a Knauer by 100-5 C-18 column (250 mm × 4.6 mm) and Knauer photodiode array detector (PAD). UHPLC analysis was performed using Knauer PAD detector with cooling autosampler and Eurospher II 100-3 C-18 column (100 mm × 3 mm) and also 2 pumps. The mean retention time, standard deviation, flow rate, and repeatability of analytical results were compared by using 2 methods. Routine runtimes in HPLC and UHPLC took 40 and 15 minutes, respectively. Changes in mobile phase composition of the 2 methods were not required. Flow rate and solvent consumption in UHPLC decreased. Diazepam and flurazepam were detected more frequently in biological samples. In UHPLC, small particle size and short length of column cause effective separation of BZs in a very short time. Reduced flow rate, solvent consumption, and injection volume cause more efficiency and less analysis costs. Thus, in the detection of BZs, UHPLC is an accurate, sensitive, and fast method with less cost of analysis.

Comparison of UHPLC and HPLC in Benzodiazepines Analysis of Postmortem Samples

PubMed Central

Behnoush, Behnam; Sheikhazadi, Ardeshir; Bazmi, Elham; Fattahi, Akbar; Sheikhazadi, Elham; Saberi Anary, Seyed Hossein

2015-01-01

Abstract The aim of this study was to compare system efficiency and analysis duration regarding the solvent consumption and system maintenance in high-pressure liquid chromatography (HPLC) and ultra high-pressure liquid chromatography (UHPLC). In a case–control study, standard solutions of 7 benzodiazepines (BZs) and 73 biological samples such as urine, tissue, stomach content, and bile that screened positive for BZs were analyzed by HPLC and UHPLC in laboratory of forensic toxicology during 2012 to 2013. HPLC analysis was performed using a Knauer by 100-5 C-18 column (250 mm × 4.6 mm) and Knauer photodiode array detector (PAD). UHPLC analysis was performed using Knauer PAD detector with cooling autosampler and Eurospher II 100-3 C-18 column (100 mm × 3 mm) and also 2 pumps. The mean retention time, standard deviation, flow rate, and repeatability of analytical results were compared by using 2 methods. Routine runtimes in HPLC and UHPLC took 40 and 15 minutes, respectively. Changes in mobile phase composition of the 2 methods were not required. Flow rate and solvent consumption in UHPLC decreased. Diazepam and flurazepam were detected more frequently in biological samples. In UHPLC, small particle size and short length of column cause effective separation of BZs in a very short time. Reduced flow rate, solvent consumption, and injection volume cause more efficiency and less analysis costs. Thus, in the detection of BZs, UHPLC is an accurate, sensitive, and fast method with less cost of analysis. PMID:25860209

Fingerprint of Hedyotis diffusa Willd. by HPLC-MS.

PubMed

Yang, Ting; Yang, Yi-Hua; Yang, Ju-Yun; Chen, Ben-Mei; Duan, Ju-Ping; Yu, Shu-Yi; Ouyang, Hong-Tao; Cheng, Jun-Ping; Chen, Yu-Xiang

2008-01-01

A HPLC-MS fingerprint method has been developed based on the consistent chromatographic features of the major chemical constituents among 10 batches of Hedyotis diffusa Willd. Chromatographic separation was conducted on a Hypersil-Keystone Hypurity C(18) column using methanol:water:acetic acid as the mobile phase. Major compounds, including oleanolic acid, ursolic acid and ferulic acid, were analysed by HPLC-MS. Their analysis was ascertained by comparison with data derived from the standard compounds. The HPLC-MS fingerprint was successfully applied to analyse and differentiate samples from different geographical origins, or processing methods. H. diffusa was well distinguished from Hedyotis chrysotricha by HPLC-MS. Therefore the establishment of fingerprint of H. diffusa is critical in assessing and controlling its overall quality.

Study of phenformin metabolism in rat liver microsomes by HPLC, CE and on-line HPLC-electrospray ionization mass spectrometry.

PubMed

Llambias, E B; Luo, J

1996-01-01

Methods for the analysis of phenformin and its metabolite by high-performance liquid chromatography (HPLC), capillary electrophoresis (CE) and high-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESIMS) are developed. The effects of pH, buffer concentration and proportion of organic modifier on the retention of the compounds in HPLC have been studied. The optimum condition was used for the separation and identification of phenformin and its metabolite in microsomal metabolism by HPLC-ESIMS. A simple CE method is also described for the separation of these compounds. Optimum incubation conditions and cofactor requirements for the formation of 4-hydroxyphenformin by microsomal preparations of rat liver were determined. A linear response in the formation of product was found with increasing concentrations of protein and up to 15 min incubation. High concentrations of phenformin inhibited its metabolite formation, and K(m) was 4 microM.

Determination of sulfonamides and trimethoprim using high temperature HPLC with simultaneous temperature and solvent gradient.

PubMed

Giegold, Sascha; Teutenberg, Thorsten; Tuerk, Jochen; Kiffmeyer, Thekla; Wenclawiak, Bernd

2008-10-01

A fast HPLC method for the analysis of eight selected sulfonamides (SA) and trimethoprim has been developed with the use of high temperature HPLC. The separation could be achieved in less than 1.5 min on a 50 mm sub 2 microm column with simultaneous solvent and temperature gradient programming. Due to the lower viscosity of the mobile phase and the increased mass transfer at higher temperatures, the separation could be performed on a conventional HPLC system obtaining peak widths at half height between 0.6 and 1.3 s.

Effect of /sup 60/Co-irradiation on penicillin G procaine in veterinary mastitis products

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tsuji, K.; Goetz, J.F.; Vanmeter, W.

The effect of /sup 60/Co-irradation on penicillin G procaine in a peanut oil-based veterinary mastitis product was examined by reversed-phase high-performance liquid chromatography (HPLC). The HPLC method is capable of separating and quantifiying procaine, penicillin G, and various degradation compounds. Values obtained by the HPLC method on the product irradiated and stored at various temperatures correlated well with those of the microbiological assay. No significant decrease in the procaine was detected even after 4.0-Mrad irradiation. The HPLC method is applicable for analysis of other beta-lactam antibiotics.

Population-level coordination of pigment response in individual cyanobacterial cells under altered nitrogen levels

DOE Office of Scientific and Technical Information (OSTI.GOV)

Murton, Jaclyn; Nagarajan, Aparna; Nguyen, Amelia Y.

Cyanobacterial phycobilisome (PBS) pigment-protein complexes harvest light and transfer the energy to reaction centers. Previous ensemble studies have shown that cyanobacteria respond to changes in nutrient availability by modifying the structure of PBS complexes, but this process has not been visualized for individual pigments at the single-cell level due to spectral overlap. We characterized the response of four key photosynthetic pigments to nitrogen depletion and repletion at the subcellular level in individual, live Synechocystis sp. PCC 6803 cells using hyperspectral confocal fluorescence microscopy and multivariate image analysis. Our results revealed that PBS degradation and re-synthesis comprise a rapid response tomore » nitrogen fluctuations, with coordinated populations of cells undergoing pigment modifications. Chlorophyll fluorescence originating from photosystem I and II decreased during nitrogen starvation, but no alteration in subcellular chlorophyll localization was found. Lastly, we observed differential rod and core pigment responses to nitrogen deprivation, suggesting that PBS complexes undergo a stepwise degradation process.« less

FOX and ETS family transcription factors regulate the pigment cell lineage in planarians.

PubMed

He, Xinwen; Lindsay-Mosher, Nicole; Li, Yan; Molinaro, Alyssa M; Pellettieri, Jason; Pearson, Bret J

2017-12-15

Many pigment cells acquire unique structural properties and gene expression profiles during animal development. The underlying differentiation pathways have been well characterized in cells formed during embryogenesis, such as the neural crest-derived melanocyte. However, much less is known about the developmental origins of pigment cells produced in adult organisms during tissue homeostasis and repair. Here we report a lineage analysis of ommochrome- and porphyrin-producing cells in the brown, freshwater planarian Schmidtea mediterranea Using an RNA-sequencing approach, we identified two classes of markers expressed in sequential fashion when new pigment cells are generated during regeneration or in response to pigment cell ablation. We also report roles for FOXF-1 and ETS-1 transcription factors, as well as for an FGFR-like molecule, in the specification and maintenance of this cell type. Together, our results provide insights into mechanisms of adult pigment cell development in the strikingly colorful Platyhelminthes phylum. © 2017. Published by The Company of Biologists Ltd.

Population-level coordination of pigment response in individual cyanobacterial cells under altered nitrogen levels

DOE PAGES

Murton, Jaclyn; Nagarajan, Aparna; Nguyen, Amelia Y.; ...

2017-07-21

Cyanobacterial phycobilisome (PBS) pigment-protein complexes harvest light and transfer the energy to reaction centers. Previous ensemble studies have shown that cyanobacteria respond to changes in nutrient availability by modifying the structure of PBS complexes, but this process has not been visualized for individual pigments at the single-cell level due to spectral overlap. We characterized the response of four key photosynthetic pigments to nitrogen depletion and repletion at the subcellular level in individual, live Synechocystis sp. PCC 6803 cells using hyperspectral confocal fluorescence microscopy and multivariate image analysis. Our results revealed that PBS degradation and re-synthesis comprise a rapid response tomore » nitrogen fluctuations, with coordinated populations of cells undergoing pigment modifications. Chlorophyll fluorescence originating from photosystem I and II decreased during nitrogen starvation, but no alteration in subcellular chlorophyll localization was found. Lastly, we observed differential rod and core pigment responses to nitrogen deprivation, suggesting that PBS complexes undergo a stepwise degradation process.« less

A smart approach to add antibacterial functionality to cellulosic pigment prints.

PubMed

Ibrahim, N A; Eid, B M; Elmaaty, T M Abou; El-Aziz, E Abd

2013-04-15

This study was devoted to enhancing the antibacterial functionality of pigment printed cotton, linen and viscose fabrics. Ag-NP's/PVP colloid, triclosan derivatives, chitosan or choline chloride was successfully incorporated into the pigment paste followed by printing and microwave curing to impart antibacterial activity to the cellulosic prints. Results obtained demonstrate that the modified pigment prints exhibit a remarkable antibacterial activity against the G+ve (Staphylococcus aureus) and G-ve (Escherichia coli) bacteria with a noticeable durability after 20 washing cycles without adversely affecting the printing and softness properties. The extent of printability and functionality of the nominated substrates are significantly governed by the type of: bio-active ingredient, binder, pigment and substrate. TEM, SEM and EDX analysis confirmed the formation of Ag-NP's/PVP colloid, of particle size range 7-14 nm, deposition of cross-linked-binder film onto the modified pigment prints, and the existence of elementary Ag and Si loaded onto fabrics surface, respectively. Copyright © 2013 Elsevier Ltd. All rights reserved.

A multi-analytical approach for the study of the pigments used in the wall paintings from a building complex on the Caelian Hill (Rome)

NASA Astrophysics Data System (ADS)

Fermo, Paola; Piazzalunga, Andrea; de Vos, Mariette; Andreoli, Martina

2013-12-01

In the present study, shards from Roman wall paintings (from the end of the first century to the fourth century A.D.) decorating the domus below the Basilica of SS. John and Paul on the Caelian Hill (Rome), were analyzed in order to identify the pigments used. The analytical techniques employed for the characterization of the pigments were the scanning electron microscope coupled with an energy dispersive spectrometer (SEM-EDS) and infrared spectroscopy (ATR and micro ATR). While SEM-EDS allowed to perform a qualitative analysis of the material, by FT-IR chemical species have been identified. The pigments identified were those mentioned in the literature for the Imperial Roman fresco painting: different types of ochre (yellow and red), mixtures containing lead, green earths and precious pigments such as cinnabar and Egyptian blue. They were often used as mixtures and the use of the most valuable pigments (cinnabar and Egyptian blue) were found in the most ancient rooms.

The Silk-protein Sericin Induces Rapid Melanization of Cultured Primary Human Retinal Pigment Epithelial Cells by Activating the NF-κB Pathway

PubMed Central

Eidet, J. R.; Reppe, S.; Pasovic, L.; Olstad, O. K.; Lyberg, T.; Khan, A. Z.; Fostad, I. G.; Chen, D. F.; Utheim, T. P.

2016-01-01

Restoration of the retinal pigment epithelial (RPE) cells to prevent further loss of vision in patients with age-related macular degeneration represents a promising novel treatment modality. Development of RPE transplants, however, requires up to 3 months of cell differentiation. We explored whether the silk protein sericin can induce maturation of primary human retinal pigment epithelial (hRPE) cells. Microarray analysis demonstrated that sericin up-regulated RPE-associated transcripts (RPE65 and CRALBP). Upstream analysis identified the NF-κB pathway as one of the top sericin-induced regulators. ELISA confirmed that sericin stimulates the main NF-κB pathway. Increased levels of RPE-associated proteins (RPE65 and the pigment melanin) in the sericin-supplemented cultures were confirmed by western blot, spectrophotometry and transmission electron microscopy. Sericin also increased cell density and reduced cell death following serum starvation in culture. Inclusion of NF-κB agonists and antagonists in the culture medium showed that activation of the NF-κB pathway appears to be necessary, but not sufficient, for sericin-induced RPE pigmentation. We conclude that sericin promotes pigmentation of cultured primary hRPE cells by activating the main NF-κB pathway. Sericin’s potential role in culture protocols for rapid differentiation of hRPE cells derived from embryonic or induced pluripotent stem cells should be investigated. PMID:26940175

The Silk-protein Sericin Induces Rapid Melanization of Cultured Primary Human Retinal Pigment Epithelial Cells by Activating the NF-κB Pathway.

PubMed

Eidet, J R; Reppe, S; Pasovic, L; Olstad, O K; Lyberg, T; Khan, A Z; Fostad, I G; Chen, D F; Utheim, T P

2016-03-04

Restoration of the retinal pigment epithelial (RPE) cells to prevent further loss of vision in patients with age-related macular degeneration represents a promising novel treatment modality. Development of RPE transplants, however, requires up to 3 months of cell differentiation. We explored whether the silk protein sericin can induce maturation of primary human retinal pigment epithelial (hRPE) cells. Microarray analysis demonstrated that sericin up-regulated RPE-associated transcripts (RPE65 and CRALBP). Upstream analysis identified the NF-κB pathway as one of the top sericin-induced regulators. ELISA confirmed that sericin stimulates the main NF-κB pathway. Increased levels of RPE-associated proteins (RPE65 and the pigment melanin) in the sericin-supplemented cultures were confirmed by western blot, spectrophotometry and transmission electron microscopy. Sericin also increased cell density and reduced cell death following serum starvation in culture. Inclusion of NF-κB agonists and antagonists in the culture medium showed that activation of the NF-κB pathway appears to be necessary, but not sufficient, for sericin-induced RPE pigmentation. We conclude that sericin promotes pigmentation of cultured primary hRPE cells by activating the main NF-κB pathway. Sericin's potential role in culture protocols for rapid differentiation of hRPE cells derived from embryonic or induced pluripotent stem cells should be investigated.

Inhibition of melanin synthesis pathway by tricyclazole increases susceptibility of Fonsecaea pedrosoi against mouse macrophages.

PubMed

Cunha, Marcel M L; Franzen, Anderson J; Alviano, Daniela S; Zanardi, Erica; Alviano, Celuta S; De Souza, Wanderley; Rozental, Sonia

2005-12-15

Fonsecaea pedrosoi produces melanin, a pigment related to virulence in pathogenic fungi. To understand the involvement of melanin in the protection of fungi, the authors used tricyclazole to inhibit the melanin pathway in F. pedrosoi. Experiments of pigmentation suggested that F. pedrosoi uniquely produces dihydroxynaphthalene-melanin. Pigments produced on cultures modified or not with tricyclazole were extracted by an alkali-acid method and submitted to infrared and ion exchange chromatography analysis; also cytochemistry analysis for cationized ferritin of whole cells was carried out. This group of experiments showed that the tricyclazole treatment on F. pedrosoi produced a melanin-like pigment, but less negatively charged and with less affinity for iron ions than that without the tricyclazole treatment, and this in turn lead to a less negatively charge cell wall surface. Scanning electron microscopy of such pigments showed that the melanin from control cultures maintained their hyphae-like structures, which have been described as "melanin-ghosts," whereas the tricyclazole pigment showed an amorphous surface. Interaction of conidia from cultures of F. pedrosoi, modified by tricyclazole or not, with peritoneal activated macrophages suggested that tricyclazole causes higher association of fungus with macrophages, weakens the fungus capacity to destroy the macrophages, and diminishes the resistance to dry fracture procedures on samples prepared for high resolution scanning electron microscopy. Copyright (c) 2005 Wiley-Liss, Inc.

Analysis of anthocyanins in commercial fruit juices by using nano-liquid chromatography-electrospray-mass spectrometry and high-performance liquid chromatography with UV-vis detector.

PubMed

Fanali, Chiara; Dugo, Laura; D'Orazio, Giovanni; Lirangi, Melania; Dachà, Marina; Dugo, Paola; Mondello, Luigi

2011-01-01

Nano-LC and conventional HPLC techniques were applied for the analysis of anthocyanins present in commercial fruit juices using a capillary column of 100 μm id and a 2.1 mm id narrow-bore C(18) column. Analytes were detected by UV-Vis at 518 nm and ESI-ion trap MS with HPLC and nano-LC, respectively. Commercial blueberry juice (14 anthocyanins detected) was used to optimize chromatographic separation of analytes and other analysis parameters. Qualitative identification of anthocyanins was performed by comparing the recorded mass spectral data with those of published papers. The use of the same mobile phase composition in both techniques revealed that the miniaturized method exhibited shorter analysis time and higher sensitivity than narrow-bore chromatography. Good intra-day and day-to-day precision of retention time was obtained in both methods with values of RSD less than 3.4 and 0.8% for nano-LC and HPLC, respectively. Quantitative analysis was performed by external standard curve calibration of cyanidin-3-O-glucoside standard. Calibration curves were linear in the concentration ranges studied, 0.1-50 and 6-50 μg/mL for HPLC-UV/Vis and nano-LC-MS, respectively. LOD and LOQ values were good for both methods. In addition to commercial blueberry juice, qualitative and quantitative analysis of other juices (e.g. raspberry, sweet cherry and pomegranate) was performed. The optimized nano-LC-MS method allowed an easy and selective identification and quantification of anthocyanins in commercial fruit juices; it offered good results, shorter analysis time and reduced mobile phase volume with respect to narrow-bore HPLC. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Simultaneous determination of potassium guaiacolsulfonate, guaifenesin, diphenhydramine HCl and carbetapentane citrate in syrups by using HPLC-DAD coupled with partial least squares multivariate calibration.

PubMed

Dönmez, Ozlem Aksu; Aşçi, Bürge; Bozdoğan, Abdürrezzak; Sungur, Sidika

2011-02-15

A simple and rapid analytical procedure was proposed for the determination of chromatographic peaks by means of partial least squares multivariate calibration (PLS) of high-performance liquid chromatography with diode array detection (HPLC-DAD). The method is exemplified with analysis of quaternary mixtures of potassium guaiacolsulfonate (PG), guaifenesin (GU), diphenhydramine HCI (DP) and carbetapentane citrate (CP) in syrup preparations. In this method, the area does not need to be directly measured and predictions are more accurate. Though the chromatographic and spectral peaks of the analytes were heavily overlapped and interferents coeluted with the compounds studied, good recoveries of analytes could be obtained with HPLC-DAD coupled with PLS calibration. This method was tested by analyzing the synthetic mixture of PG, GU, DP and CP. As a comparison method, a classsical HPLC method was used. The proposed methods were applied to syrups samples containing four drugs and the obtained results were statistically compared with each other. Finally, the main advantage of HPLC-PLS method over the classical HPLC method tried to emphasized as the using of simple mobile phase, shorter analysis time and no use of internal standard and gradient elution. Copyright © 2010 Elsevier B.V. All rights reserved.

Comparative evaluation of three different ELISA assays and HPLC-ESI-ITMS/MS for the analysis of Nε-carboxymethyl lysine in food samples.

PubMed

Gómez-Ojeda, Armando; Jaramillo-Ortíz, Sarahi; Wrobel, Katarzyna; Wrobel, Kazimierz; Barbosa-Sabanero, Gloria; Luevano-Contreras, Claudia; de la Maza, Maria Pia; Uribarri, Jaime; Del Castillo, Ma Dolores; Garay-Sevilla, Ma Eugenia

2018-03-15

N ε -carboxymethyl-lysine (CML) is measured in food, but there is a controversy concerning the most convenient yet reliable method(s) for this task. This work compares three different ELISA assays and HPLC-ESI-ITMS/MS for the analysis of CML in several food items. The four methods showed the same decreasing order of CML concentration: beef, bacon>chicken > fish>dairy products>grain products>fruits/vegetables. HPLC-ESI-ITMS/MS results highly correlated with those obtained by ELISA performed with monoclonal CML-antibody (β=0.98, p<0.0001) whereas My Bio Source® kit results were not correlated with those provided by Lamider®. Small differences of CML concentrations in food items prepared by different culinary treatment were clearly distinguished by HPLC-ESI-ITMS/MS, but could not always be detected by ELISA. This work demonstrates a reasonable relationship between CM determined by ELISA and HPLC-ESI-ITMS/MS and therefore supports the implementation of ELISA in food CML/AGEs screening. Copyright © 2017. Published by Elsevier Ltd.

Characterization and quantification of flavonoids and saponins in adzuki bean (Vigna angularis L.) by HPLC-DAD-ESI-MSn analysis.

PubMed

Liu, Rui; Cai, Zongwei; Xu, Baojun

2017-09-22

Bioactive activities of adzuki bean have been widely reported, however, the phytochemical information of adzuki bean is incomplete. The aim of this study was to characterize and quantify flavonoids and saponins in adzuki bean. High performance liquid chromatography with diode array detection and electro spray ionization-tandem multi-stage mass spectrometry (HPLC-DAD-ESI-MS n ) were applied to do qualitative and quantitative analyses. A total of 15 compounds from adzuki bean were identified by HPLC-DAD-ESI-MS n . Among 15 compounds identified, four flavonoids (catechin, vitexin-4″-O-glucoside, quercetin-3-O-glucoside, and quercetin-3-O-rutinoside) and six saponins (azukisaponin I, II, III, IV, V, and VI) in adzuki bean were further quantified by external calibration method using HPLC-MS with the program of time segment and extract ion chromatogram (EIC) analysis. Current qualitative and quantitative method based on HPLC and MS technique provides a scientific basis for in vitro and in vivo pharmacological study in the future. Graphical abstract Isolation and characterization of flavonoids and saponins from adzuki bean.

Standardization of a fluconazole bioassay and correlation of results with those obtained by high-pressure liquid chromatography.

PubMed Central

Rex, J H; Hanson, L H; Amantea, M A; Stevens, D A; Bennett, J E

1991-01-01

An improved bioassay for fluconazole was developed. This assay is sensitive in the clinically relevant range (2 to 40 micrograms/ml) and analyzes plasma, serum, and cerebrospinal fluid specimens; bioassay results correlate with results obtained by high-pressure liquid chromatography (HPLC). Bioassay and HPLC analyses of spiked plasma, serum, and cerebrospinal fluid samples (run as unknowns) gave good agreement with expected values. Analysis of specimens from patients gave equivalent results by both HPLC and bioassay. HPLC had a lower within-run coefficient of variation (less than 2.5% for HPLC versus less than 11% for bioassay) and a lower between-run coefficient of variation (less than 5% versus less than 12% for bioassay) and was more sensitive (lower limit of detection, 0.1 micrograms/ml [versus 2 micrograms/ml for bioassay]). The bioassay is, however, sufficiently accurate and sensitive for clinical specimens, and its relative simplicity, low sample volume requirement, and low equipment cost should make it the technique of choice for analysis of routine clinical specimens. PMID:1854166

Detection of pigments of halophilic endoliths from gypsum: Raman portable instrument and European Space Agency's prototype analysis

PubMed Central

Culka, Adam; Osterrothová, Kateřina; Hutchinson, Ian; Ingley, Richard; McHugh, Melissa; Oren, Aharon; Edwards, Howell G. M.; Jehlička, Jan

2014-01-01

A prototype instrument, under development at the University of Leicester, for the future European Space Agency (ESA) ExoMars mission, was used for the analysis of microbial pigments within a stratified gypsum crust from a hypersaline saltern evaporation pond at Eilat (Israel). Additionally, the same samples were analysed using a miniaturized Raman spectrometer, featuring the same 532 nm excitation. The differences in the position of the specific bands, attributed to carotenoid pigments from different coloured layers, were minor when analysed by the ESA prototype instrument; therefore, making it difficult to distinguish among the different pigments. The portable Delta Nu Advantage instrument allowed for the discrimination of microbial carotenoids from the orange/green and purple layers. The purpose of this study was to complement previous laboratory results with new data and experience with portable or handheld Raman systems, even with a dedicated prototype Raman system for the exploration of Mars. The latter is equipped with an excitation wavelength falling within the carotenoid polyene resonance region. The ESA prototype Raman instrument detected the carotenoid pigments (biomarkers) with ease, although further detailed distinctions among them were not achieved. PMID:25368354

Analysis of flavonoids and the flavonoid structural genes in brown fiber of upland cotton.

PubMed

Feng, Hongjie; Tian, Xinhui; Liu, Yongchang; Li, Yanjun; Zhang, Xinyu; Jones, Brian Joseph; Sun, Yuqiang; Sun, Jie

2013-01-01

As a result of changing consumer preferences, cotton (Gossypium Hirsutum L.) from varieties with naturally colored fibers is becoming increasingly sought after in the textile industry. The molecular mechanisms leading to colored fiber development are still largely unknown, although it is expected that the color is derived from flavanoids. Firstly, four key genes of the flavonoid biosynthetic pathway in cotton (GhC4H, GhCHS, GhF3'H, and GhF3'5'H) were cloned and studied their expression profiles during the development of brown- and white cotton fibers by QRT-PCR. And then, the concentrations of four components of the flavonoid biosynthetic pathway, naringenin, quercetin, kaempferol and myricetin in brown- and white fibers were analyzed at different developmental stages by HPLC. The predicted proteins of the four flavonoid structural genes corresponding to these genes exhibit strong sequence similarity to their counterparts in various plant species. Transcript levels for all four genes were considerably higher in developing brown fibers than in white fibers from a near isogenic line (NIL). The contents of four flavonoids (naringenin, quercetin, kaempferol and myricetin) were significantly higher in brown than in white fibers and corresponding to the biosynthetic gene expression levels. Flavonoid structural gene expression and flavonoid metabolism are important in the development of pigmentation in brown cotton fibers.

Analysis of Biomass Sugars Using a Novel HPLC Method

DOE Office of Scientific and Technical Information (OSTI.GOV)

Agblevor, F. A.; Hames, B. R.; Schell, D.

The precise quantitative analysis of biomass sugars is a very important step in the conversion of biomass feedstocks to fuels and chemicals. However, the most accurate method of biomass sugar analysis is based on the gas chromatography analysis of derivatized sugars either as alditol acetates or trimethylsilanes. The derivatization method is time consuming but the alternative high-performance liquid chromatography (HPLC) method cannot resolve most sugars found in biomass hydrolysates. We have demonstrated for the first time that by careful manipulation of the HPLC mobile phase, biomass monomeric sugars (arabinose, xylose, fructose, glucose, mannose, and galactose) can be analyzed quantitatively andmore » there is excellent baseline resolution of all the sugars. This method was demonstrated for standard sugars, pretreated corn stover liquid and solid fractions. Our method can also be used to analyze dimeric sugars (cellobiose and sucrose).« less

High-Performance Liquid Chromatography (HPLC)-Based Detection and Quantitation of Cellular c-di-GMP.

PubMed

Petrova, Olga E; Sauer, Karin

2017-01-01

The modulation of c-di-GMP levels plays a vital role in the regulation of various processes in a wide array of bacterial species. Thus, investigation of c-di-GMP regulation requires reliable methods for the assessment of c-di-GMP levels and turnover. Reversed-phase high-performance liquid chromatography (RP-HPLC) analysis has become a commonly used approach to accomplish these goals. The following describes the extraction and HPLC-based detection and quantification of c-di-GMP from Pseudomonas aeruginosa samples, a procedure that is amenable to modifications for the analysis of c-di-GMP in other bacterial species.

HPLC-based metabolic profiling and quality control of leaves of different Panax species

PubMed Central

Yang, Seung-Ok; Lee, Sang Won; Kim, Young Ock; Sohn, Sang-Hyun; Kim, Young Chang; Hyun, Dong Yoon; Hong, Yoon Pyo; Shin, Yu Su

2013-01-01

Leaves from Panax ginseng Meyer (Korean origin and Chinese origin of Korean ginseng) and P. quinquefolius (American ginseng) were harvested in Haenam province, Korea, and were analyzed to investigate patterns in major metabolites using HPLC-based metabolic profiling. Partial least squares discriminant analysis (PLS-DA) was used to analyze the HPLC chromatogram data. There was a clear separation between Panax species and/or origins from different countries in the PLS-DA score plots. The ginsenoside compounds of Rg1, Re, Rg2, Rb2, Rb3, and Rd in Korean leaves were higher than in Chinese and American ginseng leaves, and the Rb1 level in P. quinquefolius leaves was higher than in P. ginseng (Korean origin or Chinese origin). HPLC chromatogram data coupled with multivariate statistical analysis can be used to profile the metabolite content and undertake quality control of Panax products. PMID:23717177

Development of a percutaneous penetration predictive model by SR-FTIR.

PubMed

Jungman, E; Laugel, C; Rutledge, D N; Dumas, P; Baillet-Guffroy, A

2013-01-30

This work focused on developing a new evaluation criterion of percutaneous penetration, in complement to Log Pow and MW and based on high spatial resolution Fourier transformed infrared (FTIR) microspectroscopy with a synchrotron source (SR-FTIR). Classic Franz cell experiments were run and after 22 h molecule distribution in skin was determined either by HPLC or by SR-FTIR. HPLC data served as reference. HPLC and SR-FTIR results were compared and a new predictive criterion based from SR-FTIR results, named S(index), was determined using a multi-block data analysis technique (ComDim). A predictive cartography of the distribution of molecules in the skin was built and compared to OECD predictive cartography. This new criterion S(index) and the cartography using SR-FTIR/HPLC results provides relevant information for risk analysis regarding prediction of percutaneous penetration and could be used to build a new mathematical model. Copyright © 2012 Elsevier B.V. All rights reserved.

Development and validation of high-performance liquid chromatography and high-performance thin-layer chromatography methods for the quantification of khellin in Ammi visnaga seed

PubMed Central

Kamal, Abid; Khan, Washim; Ahmad, Sayeed; Ahmad, F. J.; Saleem, Kishwar

2015-01-01

Objective: The present study was used to design simple, accurate and sensitive reversed phase-high-performance liquid chromatography RP-HPLC and high-performance thin-layer chromatography (HPTLC) methods for the development of quantification of khellin present in the seeds of Ammi visnaga. Materials and Methods: RP-HPLC analysis was performed on a C18 column with methanol: Water (75: 25, v/v) as a mobile phase. The HPTLC method involved densitometric evaluation of khellin after resolving it on silica gel plate using ethyl acetate: Toluene: Formic acid (5.5:4.0:0.5, v/v/v) as a mobile phase. Results: The developed HPLC and HPTLC methods were validated for precision (interday, intraday and intersystem), robustness and accuracy, limit of detection and limit of quantification. The relationship between the concentration of standard solutions and the peak response was linear in both HPLC and HPTLC methods with the concentration range of 10–80 μg/mL in HPLC and 25–1,000 ng/spot in HPTLC for khellin. The % relative standard deviation values for method precision was found to be 0.63–1.97%, 0.62–2.05% in HPLC and HPTLC for khellin respectively. Accuracy of the method was checked by recovery studies conducted at three different concentration levels and the average percentage recovery was found to be 100.53% in HPLC and 100.08% in HPTLC for khellin. Conclusions: The developed HPLC and HPTLC methods for the quantification of khellin were found simple, precise, specific, sensitive and accurate which can be used for routine analysis and quality control of A. visnaga and several formulations containing it as an ingredient. PMID:26681890

Bacteria entombed in the center of cholesterol gallstones induce fewer infectious manifestations than bacteria in the matrix of pigment stones.

PubMed

Stewart, Lygia; Griffiss, J McLeod; Jarvis, Gary A; Way, Lawrence W

2007-10-01

The clinical significance of bacteria in the pigment centers of cholesterol stones is unknown. We compared the infectious manifestations and characteristics of bacteria from pigment stones and predominantly cholesterol stones. Three hundred forty patients were studied. Bile was cultured. Gallstones were cultured and examined with scanning electron microscopy. Level of bacterial immunoglobulin G (bile, serum), complement killing, and tumor necrosis factor-alpha production were determined. Twenty-three percent of cholesterol stones and 68% of pigment stones contained bacteria (P < 0.0001). Stone culture correlated with scanning electron microscopy results. Pigment stone bacteria were more often present in bile and blood. Cholesterol stone bacteria caused more severe infections (19%) than sterile stones (0%), but less than pigment stone bacteria (57%) (P < 0.0001). Serum and bile from patients with cholesterol stone bacteria had less bacterial-specific immunoglobulin G. Cholesterol stone bacteria produced more slime. Pigment stone bacteria were more often killed by a patient's serum. Tumor necrosis factor-alpha production of the groups was similar. Bacteria are readily cultured from cholesterol stones with pigment centers, allowing for analysis of their virulence factors. Bacteria sequestered in cholesterol stones cause infectious manifestations, but less than bacteria in pigment stones. Possibly because of their isolation, cholesterol stone bacteria were less often present in bile and blood, induced less immunoglobulin G, were less often killed by a patient's serum, and demonstrated fewer infectious manifestations than pigment stone bacteria. This is the first study to analyze the clinical relevance of bacteria within cholesterol gallstones.

Taxonomy, virulence and epidemiology of black-pigmented Bacteroides species in relation to oral infections.

PubMed

van Steenbergen, T J; van Winkelhoff, A J; van der Velden, U; de Graaff, J

1989-01-01

Black-pigmented Bacteroides species are recognized as suspected pathogens of oral infections. Developments in the taxonomy of this group include description of a new asaccharolytic species, Bacteroides salivosus, and proposal for the reclassification of the asaccharolytic species into a separate genus, Porphyromonas. Studies on the pathogenicity and virulence of black-pigmented Bacteroides species have identified Bacteroides gingivalis as the most virulent species. B. gingivalis and Bacteroides intermedius have been associated with periodontal diseases; Bacteroides endodontalis is isolated specifically from infections in the oral cavity, and other black-pigmented Bacteroides species are recovered from oral mucous sites. DNA restriction endonuclease analysis was adapted for typing of B. gingivalis and B. intermedius.

Spectroscopic characterization and quantitative determination of atorvastatin calcium impurities by novel HPLC method

NASA Astrophysics Data System (ADS)

Gupta, Lokesh Kumar

2012-11-01

Seven process related impurities were identified by LC-MS in the atorvastatin calcium drug substance. These impurities were identified by LC-MS. The structure of impurities was confirmed by modern spectroscopic techniques like 1H NMR and IR and physicochemical studies conducted by using synthesized authentic reference compounds. The synthesized reference samples of the impurity compounds were used for the quantitative HPLC determination. These impurities were detected by newly developed gradient, reverse phase high performance liquid chromatographic (HPLC) method. The system suitability of HPLC analysis established the validity of the separation. The analytical method was validated according to International Conference of Harmonization (ICH) with respect to specificity, precision, accuracy, linearity, robustness and stability of analytical solutions to demonstrate the power of newly developed HPLC method.

A double-blind, placebo-controlled study on the effects of lutein and zeaxanthin on photostress recovery, glare disability, and chromatic contrast.

PubMed

Hammond, Billy R; Fletcher, Laura M; Roos, Franz; Wittwer, Jonas; Schalch, Wolfgang

2014-12-02

Past studies have shown that higher macular pigment optical density (MPOD) and lutein (L) and zeaxanthin (Z) supplementation are related to improvements in glare disability, photostress recovery, and chromatic contrast. This study assessed those links using a randomized, double-blind, placebo-controlled design. The visual effects of 1 year of supplementing L (10 mg/d) and Z (2 mg/d) were investigated. One hundred fifteen young, healthy subjects were recruited and randomized into the study (58 received placebo, 57 L+Z). Several dependent measures were collected at baseline and then once every 3 months: serum L and Z measured by HPLC chromatography; MPOD measured using customized heterochromatic flicker photometry; photostress recovery assessed by measuring the time needed to recover visual acquisition of a grating target after 30 seconds of an intense xenon white flash exposure; glare disability evaluated as the energy in a surrounding annulus necessary to veil a central grating target; and chromatic contrast assessed by measuring thresholds for a yellow grating target superposed on a 460-nm background. Macular pigment optical density increased significantly versus placebo at all eccentricities (10, 30, 60, and 105 minutes from the center of the macula). Serum L and Z also increased significantly by the first follow-up visit (at 3 months), and remained elevated throughout the intervention period of 1 year. Chromatic contrast and photostress recovery time improved significantly versus placebo. Glare disability was correlated with macular pigment density throughout the study period but did not increase significantly in the treated group. Daily supplementation with L+Z resulted in significant increase in serum levels and MPOD and improvements in chromatic contrast and recovery from photostress. These results are consistent with past studies showing that increasing MPOD leads to improved visual performance. (ClinicalTrials.gov number, NCT00909090.). Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.

Effects of atropine on refractive development, dopamine release, and slow retinal potentials in the chick.

PubMed

Schwahn, H N; Kaymak, H; Schaeffel, F

2000-01-01

Atropine has previously been found to suppress visually induced myopia both in animals and humans. The mechanism of its action is unclear. We have studied its retinal effects in an in vitro preparation, using the retina-pigment epithelium-choroid complex of the chick eye. In vivo, deprivation myopia was induced by translucent goggles. Atropine solution was injected into the vitreous at two-day intervals. Dopamine release from the retina following atropine injection in vivo and from the in vitro retina preparation was quantified by HPLC-EC. In vitro preparations of the isolated chick retina-pigment epithelium-choroid were superfused with atropine. Light-induced potentials (local ERG), slow standing potentials from the retinal pigment epithelium/neural retina, and extracellular potassium concentrations were recorded. In line with previous findings, intravitreal injections of atropine (25 microg, 250 microg) reduced deprivation myopia in a dose-dependent manner. Atropine increased the release of the neurotransmitter dopamine into the superfusate in vitro at 100-500 microM and into the vitreous in vivo at 250 microg. Before an increase was measured in the vitreous, the retinal dopamine content was elevated. In concentrations equivalent to the intravitreal concentration to suppress myopia in vivo (200-800 microM), atropine induced spreading depression (SD) in the in vitro preparation. In contrast, muscarinic agonists, acetylcholine and pilocarpine, did not induce SD. Atropine reduced the ERG b- and d-wave, led to damped oscillations of RPE potentials, and reversed the ERG c-wave. Atropine suppressed myopia only at doses at which severe nonspecific side effects were observed in the retina. Atropine seems to intrude massively into the vital functions of the retina as indicated by the occurrence of SD. We conclude that atropine, by inducing SD, boosts neurotransmitter release from cellular stores, which may cancel out a presumed retinal signal that controls eye growth and through this, myopia.

Lake and Sea Populations of Mysis relicta (Crustacea, Mysida) with Different Visual-Pigment Absorbance Spectra Use the Same A1 Chromophore

PubMed Central

Belikov, Nikolai; Yakovleva, Marina; Feldman, Tatiana; Demina, Olga; Khodonov, Andrei; Lindström, Magnus; Donner, Kristian; Ostrovsky, Mikhail

2014-01-01

Glacial-relict species of the genus Mysis (opossum shrimps) inhabiting both fresh-water lakes and brackish sea waters in northern Europe show a consistent lake/sea dichotomy in eye spectral sensitivity. The absorbance peak (λmax) recorded by microspectrophotometry in isolated rhabdoms is invariably 20–30 nm red-shifted in “lake” compared with “sea” populations. The dichotomy holds across species, major opsin lineages and light environments. Chromophore exchange from A1 to A2 (retinal → 3,4-didehydroretinal) is a well-known mechanism for red-shifting visual pigments depending on environmental conditions or stages of life history, present not only in fishes and amphibians, but in some crustaceans as well. We tested the hypothesis that the lake/sea dichotomy in Mysis is due to the use of different chromophores, focussing on two populations of M. relicta from, respectively, a Finnish lake and the Baltic Sea. They are genetically very similar, having been separated for less than 10 kyr, and their rhabdoms show a typical lake/sea difference in λmax (554 nm vs. 529 nm). Gene sequencing has revealed no differences translating into amino acid substitutions in the transmembrane parts of their opsins. We determined the chromophore identity (A1 or A2) in the eyes of these two populations by HPLC, using as standards pure chromophores A1 and A2 as well as extracts from bovine (A1) and goldfish (A2) retinas. We found that the visual-pigment chromophore in both populations is A1 exclusively. Thus the spectral difference between these two populations of M. relicta is not due to the use of different chromophores. We argue that this conclusion is likely to hold for all populations of M. relicta as well as its European sibling species. PMID:24516590

Seed Carotenoid and Tocochromanol Composition of Wild Fabaceae Species Is Shaped by Phylogeny and Ecological Factors

PubMed Central

Fernández-Marín, Beatriz; Míguez, Fátima; Méndez-Fernández, Leire; Agut, Agustí; Becerril, José M.; García-Plazaola, José I.; Kranner, Ilse; Colville, Louise

2017-01-01

Carotenoids distribution and function in seeds have been very scarcely studied, notwithstanding their pivotal roles in plants that include photosynthesis and phytohormone synthesis, pigmentation, membrane stabilization and antioxidant activity. Their relationship with tocochromanols, whose critical role in maintaining seed viability has already been evidenced, and with chlorophylls, whose retention in mature seed is thought to have negative effects on storability, remain also unexplored. Here, we aimed at elucidating seed carotenoids relationship with tocochromanols and chlorophylls with regard to phylogenetic and ecological traits and at understanding their changes during germination. The composition and distribution of carotenoids were investigated in seeds of a wide range of wild species across the Fabaceae (the second-most economically important family after the Poaceae). Photosynthetic pigments and tocochromanols were analyzed by HPLC in mature dry seeds of 50 species representative of 5 subfamilies within the Fabaceae (including taxa that represent all continents, biomes and life forms within the family) and at key timepoints during seedling establishment in three species representative of distinct clades. Total-carotenoids content positively correlated with tocopherols in the basal subfamilies Detarioideae, Cercidoideae, and Dialioideae, and with chlorophylls in the Papilionoideae. Papilionoideae lacked tocotrienols and had the highest total-carotenoids, chlorophyll and γ-tocopherol contents. Interestingly, lutein epoxide was present in 72% of the species including several herbs from different subfamilies. Overall, species original from temperate biomes presented higher carotenoids and lower tocochromanols levels than those from tropical biomes. Also shrub species showed higher carotenoids content than herbs and trees. During germination, total content of photosynthetic pigments increased in parallel to changes in relative abundance of carotenoids: zeaxanthin and anteraxanthin decreased and β-carotene augmented. Notably, the highest contents of nutritionally valuable carotenoids were found in Papilionoideae subfamily to which all pulses of socio-economic importance belong. The major differences in carotenoids and tocochromanols composition across the Fabaceae are apparently related to phylogeny in conjunction with ecological traits such as biome and growth form. PMID:28883825

Sustained Release Oral Nanoformulated Green Tea for Prostate Cancer Prevention

DTIC Science & Technology

2012-05-01

31.25 ng/ml. The calibration samples were measured twice. The HPLC was a Shimadzu Prominence LC system containing a CBM-20A system controller...MDS SCIEX Ontario, Canada) equipped with a Turbo V Source and Turbo Ion Spray was coupled to the HPLC . The mass spectrometer was operated in...treated mice were subjected to HPLC and GC mass spectrometry for analysis of pharmacokinetic distribution of EGCG and bioavailability. Though we have

Carotenoid composition in the fruits of red paprika (Capsicum annuum var. lycopersiciforme rubrum) during ripening; biosynthesis of carotenoids in red paprika.

PubMed

Deli, J; Molnár, P; Matus, Z; Tóth, G

2001-03-01

The changes in the carotenoid pigments of the Capsicum annuum var. lycopersiciforme rubrum during maturation have been investigated quantitatively by means of a HPLC technique. In all of the chromatograms, 40 peaks were detected; 34 carotenoids were identified. The total carotenoid content of the ripe fruits was about 1.3 g/100 g of dry weight, of which capsanthin constituted 37%, zeaxanthin was 8%, cucurbitaxanthin A was 7%, capsorubin constituted 3.2%, and beta-carotene accounted for 9%. The remainder was composed of capsanthin 5,6-epoxide, capsanthin 3,6-epoxide, 5,6-diepikarpoxanthin, violaxanthin, antheraxanthin, beta-cryptoxanthin, and several cis isomers and furanoid oxides. The possible biosynthetic routes for the formation of minor carotenoids containing 3,5,6-trihydroxy-beta-, 3,6-epoxy-beta-, and 6-hydroxy-gamma-end groups are described.

Electronic structure and optical properties of some anthocyanins extracted from grapes

NASA Astrophysics Data System (ADS)

Iosub, Ion; Kajzar, Francois; Makowska-Janusik, Malgorzata; Meghea, Aurelia; Tane, Alexandrina; Rau, Ileana

2012-08-01

The pallet of applications of natural dyes and pigments is continuously extending as a stringent need to meet the challenges arisen from sustainable development, particularly related to replacing synthetic chemicals by eco-friendly renewable raw materials. In this respect, the well known nutritive and therapeutic properties demonstrated by anthocyanin compounds present in selective plant extracts of numerous fruits, vegetables and flowers could be largely complemented by their characteristics as chromophores and fluorophores with high potential in promising applications in bio-imaging for diagnostic and therapy, but also in electronic and photonic devices, solar cells, etc. In this context, this paper proposes an improved protocol for extraction and HPLC detection of anthocyanins from black grapes. Moreover, in order to exploit their potential to be applied as chromatic and fluorescence molecular probes, the electronic properties have been studied by correlating spectral characteristics with trichromatic parameters, combined with quantum chemistry computation.

Pigment analysis by Raman microscopy and portable X-ray fluorescence (pXRF) of thirteenth to fourteenth century illuminations and cuttings from Bologna

PubMed Central

Clark, Robin J. H.; Jones, Richard; Gibbs, Robert

2016-01-01

Non-destructive pigment analysis by Raman microscopy (RM) and portable X-ray fluorescence (pXRF) has been carried out on some Bolognese illuminations and cuttings chosen to represent the beginnings, evolution and height of Bolognese illuminated manuscript production. Dating to the thirteenth and fourteenth centuries and held in a private collection, the study provides evidence for the pigments generally used in this period. The results, which are compared with those obtained for other north Italian artwork, show the developments in usage of artistic materials and technique. Also addressed in this study is an examination of the respective roles of RM and pXRF analysis in this area of technical art history. This article is part of the themed issue ‘Raman spectroscopy in art and archaeology’. PMID:27799427

Stability-Indicating Related Substances HPLC Method for Droxidopa and Characterization of Related Substances Using LC-MS and NMR.

PubMed

Kumar, Thangarathinam; Ramya, Mohandass; Arockiasamy Xavier, S J

2016-11-01

Stress degradation studies using high-performance liquid chromatography (HPLC) was performed and validated for Droxidopa (L-DOPS). Droxidopa was susceptible to acid hydrolysis (0.1 N HCl), alkaline hydrolysis (0.15 N NaOH) and thermal degradation (105°C). It was found to be resistant to white light, oxidation and UV light exposure (72 h). The thermal, acid and alkali degradation impurities were detected with the retention time (RT) of 12.7, 19.25 and 22.95 min. Our HPLC method detected process impurities (2R,3R)-2-amino-3-(3,4-dihydroxyphenyl)-3-hydroxypropionic acid (Impurity H), N-Hydroxypthalimide (Impurity N), (2R,3S)-2-amino-3-(benzo[d][1,3]dioxol-5-yl)-3-hydroxypropionic acid (Impurity L) and L-threo n-phthaloyl-3-(3, 4-dihydroxyphenyl)-serine (Intermediate) with RTs of 3.48, 15.5, 25.76 and 28.0 min. The related substances were further characterized and confirmed by liquid chromatography-mass spectroscopy (LC-MS), and nuclear magnetic resonance spectroscopy analysis. Our HPLC method detected up to 0.05 µg/mL of Droxidopa with S/N > 3.0 and quantified up to 0.10 µg /mL of Droxidopa with S/N ratio > 10.0. Droxidopa was highly stable for 12 h after its preparation for HPLC analysis. Our newly developed HPLC method was highly precise, specific, reliable and accurate for the analysis of Droxidopa and its related substances. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Reverse-phase HPLC analysis of human alpha crystallin.

PubMed

Swamy, M S; Abraham, E C

1991-03-01

A rapid and highly sensitive reverse-phase HPLC (RP-HPLC) method was used to separate crystallin subunits from human alpha crystallin. Three distinct peaks were separated; by electrophoretic and immunological analyses the first and second peaks were identified as alpha B and alpha A respectively. On the other hand, peak 3 appeared to be a modified form of alpha crystallin. The ratio of alpha A and alpha B proteins was 3:1 in 1 day old lenses which gradually changed to 2:1 in 17 year old lenses and to 1:1 in the 50 and 82 year old whole lenses and 82 year old lens cortex, with a concomitant increase in the modified alpha, suggesting that alpha A subunits are relatively more involved in aggregation. Analysis of the 82 year old lens nucleus also supported this conclusion. The RP-HPLC analysis of the HMW aggregate fraction showed substantial enrichment of the modified alpha. The alpha A and alpha B subunits independently reassociated to form polymeric alpha crystallin whereas the modified alpha reassociated to form HMW aggregates as shown by molecular sieve HPLC. Hence it appears that the HMW aggregate peak was constituted by modified alpha crystallin. Only in the peak 3 material the 280 nm absorbance was about 2-fold higher than what was expected from the actual protein content. The data suggest that the changes induced by post-translational modifications may have some role in the formation of modified alpha. The present RP-HPLC method is useful in separating these modified alpha from the unmodified alpha A and alpha B subunits.

High-performance liquid chromatography-mass spectrometry for mapping and sequencing glycosaminoglycan-derived oligosaccharides

PubMed Central

Volpi, Nicola; Linhardt, Robert J

2012-01-01

Glycosaminoglycans (GAGs) have proven to be very difficult to analyze and characterize because of their high negative charge density, polydispersity and sequence heterogeneity. As the specificity of the interactions between GAGs and proteins results from the structure of these polysaccharides, an understanding of GAG structure is essential for developing a structure–activity relationship. Electrospray ionization (ESI) mass spectrometry (MS) is particularly promising for the analysis of oligosaccharides chemically or enzymatically generated by GAGs because of its relatively soft ionization capacity. Furthermore, on-line high-performance liquid chromatography (HPLC)-MS greatly enhances the characterization of complex mixtures of GAG-derived oligosaccharides, providing important structural information and affording their disaccharide composition. A detailed protocol for producing oligosaccharides from various GAGs, using controlled, specific enzymatic or chemical depolymerization, is presented, together with their HPLC separation, using volatile reversed-phase ion-pairing reagents and on-line ESI-MS structural identification. This analysis provides an oligosaccharide map together with sequence information from a reading frame beginning at the nonreducing end of the GAG chains. The preparation of oligosaccharides can be carried out in 10 h, with subsequent HPLC analysis in 1–2 h and HPLC-MS analysis taking another 2 h. PMID:20448545

The Effect of maceration period on contents and color brightness of phycoerythrin from Gracilaria sp.

NASA Astrophysics Data System (ADS)

Lidiana, H.; Sulmartiwi, L.; Andriyono, S.

2018-04-01

Natural pigment from seaweed, is currently required by humans as one of active compounds that are usefull in the field of health, cosmetics and food. One of the pigments which potentially can be developed is red phycobilin pigment. The pigment can be obtained from Gracilaria sp. throught maceration method. This research aims to find out wheter there is influence from the long maceration time toward to the content and the phycoerytrin color brightness of Gracilaria sp. The reseach method used experimental experiment with complete randomized design (CRD) which was consisted of nine treatments of long time maceration period three replications. Data analysis was used the Analysis of Variance (ANOVA) and continued to Duncan’s Multiple Distance Test to. The result showed that treatments with the time of maceration for 30 hours was the best treatment on phycoerytrin content about 0.98 mg/g with the brightness value was 19.73 L*, reddish value was 7.23 a*, and yellowish value was 2.87 b* of Gracilaria sp.

Non-invasive and micro-destructive investigation of the Domus Aurea wall painting decorations.

PubMed

Clementi, Catia; Ciocan, Valeria; Vagnini, Manuela; Doherty, Brenda; Tabasso, Marisa Laurenzi; Conti, Cinzia; Brunetti, Brunetto Giovanni; Miliani, Costanza

2011-10-01

The paper reports on the exploitation of an educated multi-technique analytical approach based on a wide non invasive step followed by a focused micro-destructive step, aimed at the minimally invasive identification of the pigments decorating the ceiling of the Gilded Vault of the Domus Aurea in Rome. The combination of elemental analysis with molecular characterization provided by X-ray fluorescence and UV-vis spectroscopies, respectively, allowed for the in situ non-invasive identification of a remarkable number of pigments, namely Egyptian blue, green earth, cinnabar, red ochre and an anthraquinonic lake. The study was completed with the Raman analysis of two bulk samples, in particular, SERS measurements allowed for the speciation of the anthraquinonic pigment. Elemental mapping by scanning electron microscopy-energy dispersive spectrometer combined with micro-fluorimetry on cross-section gave an insight into both the distribution of different blend of pigments and on the nature of the inorganic support of the red dye.

Persistent and widespread occurrence of bioactive quinone pigments during post-Paleozoic crinoid diversification

PubMed Central

Wolkenstein, Klaus

2015-01-01

Secondary metabolites often play an important role in the adaptation of organisms to their environment. However, little is known about the secondary metabolites of ancient organisms and their evolutionary history. Chemical analysis of exceptionally well-preserved colored fossil crinoids and modern crinoids from the deep sea suggests that bioactive polycyclic quinones related to hypericin were, and still are, globally widespread in post-Paleozoic crinoids. The discovery of hypericinoid pigments both in fossil and in present-day representatives of the order Isocrinida indicates that the pigments remained almost unchanged since the Mesozoic, also suggesting that the original color of hypericinoid-containing ancient crinoids may have been analogous to that of their modern relatives. The persistent and widespread occurrence, spatially as well as taxonomically, of hypericinoid pigments in various orders during the adaptive radiation of post-Paleozoic crinoids suggests a general functional importance of the pigments, contributing to the evolutionary success of the Crinoidea. PMID:25730856

Effect of ascorbic acid on the stability of beta-carotene and capsanthin in paprika (Capsicum annuum) powder.

PubMed

Morais, H; Rodrigues, P; Ramos, C; Forgács, E; Cserháti, T; Oliveira, J

2002-10-01

The effect of ascorbic acid, light, and storage on the stability of the pigments beta-carotene and capsanthin in red pepper (Capsicum annuum) powder has been elucidated by determining the amount of pigment in samples treated by various concentrations of ascorbic acid. Determination of pigment concentration has been performed after different storage times using high-performance liquid chromatography. The dependence of the concentration of pigments on the concentration of ascorbic acid, presence of light and the storage time has been assessed by stepwise regression analysis. The concentration of pigments decreased at longer storage time and increased at higher concentration of ascorbic acid, beta-carotene being more sensitive towards storage time and concentration of ascorbic acid than capsanthin. Interaction between the effects of light and storage time, and light and concentration of ascorbic acid has been established.

Detection of pigment network in dermatoscopy images using texture analysis

PubMed Central

Anantha, Murali; Moss, Randy H.; Stoecker, William V.

2011-01-01

Dermatoscopy, also known as dermoscopy or epiluminescence microscopy (ELM), is a non-invasive, in vivo technique, which permits visualization of features of pigmented melanocytic neoplasms that are not discernable by examination with the naked eye. ELM offers a completely new range of visual features. One such prominent feature is the pigment network. Two texture-based algorithms are developed for the detection of pigment network. These methods are applicable to various texture patterns in dermatoscopy images, including patterns that lack fine lines such as cobblestone, follicular, or thickened network patterns. Two texture algorithms, Laws energy masks and the neighborhood gray-level dependence matrix (NGLDM) large number emphasis, were optimized on a set of 155 dermatoscopy images and compared. Results suggest superiority of Laws energy masks for pigment network detection in dermatoscopy images. For both methods, a texel width of 10 pixels or approximately 0.22 mm is found for dermatoscopy images. PMID:15249068

Pigment Production Analysis in Human Melanoma Cells.

PubMed

Hopkin, Amelia Soto; Paterson, Elyse K; Ruiz, Rolando; Ganesan, Anand K

2016-05-25

The human epidermal melanocyte is a highly specialized pigmented cell that serves to protect the epidermis from ultraviolet (UV) damage through the production of melanin, or melanogenesis. Misregulation in melanogenesis leading to either hyper- or hypo-pigmentation is found in human diseases such as malasma and vitiligo. Current therapies for these diseases are largely unsuccessful and the need for new therapies is necessary. In order to identify genes and or compounds that can alter melanogenesis, methods are required that can detect changes in pigment production as well as expression of key melanogenesis transcription factors and enzymes. Here we describe methods to detect changes in melanogenesis in a human melanoma cell line, MNT-1, by (1) analyzing pigment production by measuring the absorbance of melanin present by spectrophotometry, (2) analyzing transcript expression of potent regulators of melanogenesis by qunatitative reverse-transcription (RT)PCR and (3) analyzing protein expression of potent regulators of melanogenesis by Western blot (WB).

Color stabilization of red wines. A chemical and colloidal approach.

PubMed

Alcalde-Eon, Cristina; García-Estévez, Ignacio; Puente, Victor; Rivas-Gonzalo, Julián C; Escribano-Bailón, M Teresa

2014-07-23

The effects of cold treatment and time on CIELAB color parameters and on anthocyanin and anthocyanin-derived pigments composition have been evaluated as has been the effectiveness of either an enological tannin or a mannoprotein (M) on their stabilization. With respect to color, hue (hab) was increased in the wines treated with both enological products. Furthermore, the color changes induced by cold treatment were lessened by the addition of these two enological products, although the protective effect was higher for the wines treated with M. The pigment analysis revealed higher percentages of anthocyanin-derived pigments in tannin and M-treated samples (in both cold treated and not) in relation to control ones. The addition of the enological tannin may favor the synthesis of anthocyanin-derived pigments, which are chemically more stable than native anthocyanins, whereas M seems to stabilize anthocyanin-derived pigments from a colloidal point of view, avoiding their aggregation and further precipitation.

Color characterization of coatings with diffraction pigments.

PubMed

Ferrero, A; Bernad, B; Campos, J; Perales, E; Velázquez, J L; Martínez-Verdú, F M

2016-10-01

Coatings with diffraction pigments present high iridescence, which needs to be characterized in order to describe their appearance. The spectral bidirectional reflectance distribution functions (BRDFs) of six coatings with SpectraFlair diffraction pigments were measured using the robot-arm-based goniospectrophotometer GEFE, designed and developed at CSIC. Principal component analysis has been applied to study the coatings of BRDF data. From data evaluation and based on theoretical considerations, we propose a relevant geometric factor to study the spectral reflectance and color gamut variation of coatings with diffraction pigments. At fixed values of this geometric factor, the spectral BRDF component due to diffraction is almost constant. Commercially available portable goniospectrophotometers, extensively used in several industries (automotive and others), should be provided with more aspecular measurement angles to characterize the complex reflectance of goniochromatic coatings based on diffraction pigments, but they would not require either more than one irradiation angle or additional out-of-plane geometries.

Analysis of major elements in pigmented melanocytic chicken skin using laser-induced breakdown spectroscopy.

PubMed

Lee, Jong Jin; Moon, Youngmin; Han, Jung Hyun; Jeong, Sungho

2017-04-01

The concentration difference of major elements in melanocytic skin with respect to pigmentation level is analysed by laser-induced breakdown spectroscopy (LIBS) to investigate the applicability of LIBS as an in situ feedback tool for selective and complete laser removal of melanocytic skin tissue like nevus. The skin of black silkie chicken which had a characteristic darkly pigmented perifollicular skin surrounded by lightly pigmented extrafollicular skin was used as the sample. The results showed higher LIBS signal intensities of Ca 2+ and Mg 2+ but lower intensities of Na + , Cl - and K + in the perifollicular skin than in the extrafollicular skin, which demonstrated the feasibility to use LIBS as a reliable method to distinguish skin tissues with difference in pigmentation level. Plasma emission of biochemical elements generated with a laser irradiation on melanocytic skin lesion. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Recent advances in micro-scale and nano-scale high-performance liquid-phase chromatography for proteome research.

PubMed

Tao, Dingyin; Zhang, Lihua; Shan, Yichu; Liang, Zhen; Zhang, Yukui

2011-01-01

High-performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS-MS) is regarded as one of the most powerful techniques for separation and identification of proteins. Recently, much effort has been made to improve the separation capacity, detection sensitivity, and analysis throughput of micro- and nano-HPLC, by increasing column length, reducing column internal diameter, and using integrated techniques. Development of HPLC columns has also been rapid, as a result of the use of submicrometer packing materials and monolithic columns. All these innovations result in clearly improved performance of micro- and nano-HPLC for proteome research.

Primary structure of an adipokinetic neuropeptide from the rhinoceros beetle, Oryctes rhinoceros L (Coleoptera: Dynastidae).

PubMed

Ajay Kumar, A P; Gokuldas, M

2011-07-01

Neuropeptides play an important role in cellular communication in vertebrates. This is also true for insects in which many physiological, developmental and behavioral processes are affected by neuropeptides produced in neurosecretory cells of the retrocerebral complex. Small neuropeptides of the adipokinetic hormone/red pigment concentrating hormone family (AKH/RPCH) are one of the important groups of peptides that regulate physiological homeostasis. The present investigation was carried out to elucidate the primary structure of adipokinetic neuropeptides in the rhinoceros beetle, O. rhinoceros. In the present investigation, an adipokinetic neuropeptide from the coconut pest, Oryctes rhinoceros was isolated from corpora cardiaca by HPLC; the chromatographic fractions were tested for adipokinetic activity in the plant bug, Iphita limbata in vivo. Two UV absorbance peaks were found to be significantly active in elevating haemolymph lipid levels. MALDI-MS analysis of the extract indicated that the molecular mass, 1003.70 Da is similar to the already known AKH from another beetle, Melolontha melolontha. MALDI-MS/MS analysis confirmed that its primary structure is exactly similar to the structure reported for the Melme-AKH (pE-L-N-Y-S-P-D-W-NH2). The findings suggest that the distribution of AKH peptides has shown that there exists a taxonomic order or family specificity. This data can be used as additional information to aid in the construction of phylogenetic trees by means of computer programme and protein parsimony algorithms.

Seasonal changes in the sensitivity of river microalgae to atrazine and isoproturon along a contamination gradient.

PubMed

Dorigo, Ursula; Bourrain, Xavier; Bérard, Annette; Leboulanger, Christophe

2004-01-05

A study was undertaken to investigate the environmental impact of herbicides on natural communities of freshwater periphyton and phytoplankton in the river Ozanne and in related nearby water reservoirs, including both pristine and pesticide- (atrazine and isoproturon) contaminated stations. The microalgal toxicity of both herbicides was investigated by short-term studies, using the in vivo fluorescence pattern to perform dose-effect experiments. The taxonomic composition of the communities sampled was assessed by microscopy and by HPLC pigment analysis. The EC50 (periphyton) or EC125 (phytoplankton) values, calculated using in vivo fluorescence endpoints, increased with the herbicide concentration found in the water. In contrast, the structure of the algal communities (periphyton) inhabiting the contaminated stations seemed to be permanently affected when compared to the reference community. A 'memory effect' could be detected, both in herbicide sensitivity and in the structure of periphytic communities that persisted even when peak contaminations had disappeared. This study shows that the response of algal communities is likely to reflect past selection pressures, and suggests that the function and structure of a community could both be modified by the persistent or repeated presence of microcontaminants in natural environments. We could use short-term ecotoxicological tests on freshwater microalgae to assess the effects of past temporary contaminations by agricultural pesticides, and combining this with diversity indices could make it possible to assess the ecotoxicological risk of herbicide contamination even when a complete chemical analysis of the contamination is not feasible.

Erwinia gerundensis sp. nov., a cosmopolitan epiphyte originally isolated from pome fruit trees.

PubMed

Rezzonico, Fabio; Smits, Theo H M; Born, Yannick; Blom, Jochen; Frey, Jürg E; Goesmann, Alexander; Cleenwerck, Ilse; de Vos, Paul; Bonaterra, Anna; Duffy, Brion; Montesinos, Emilio

2016-03-01

A survey to obtain potential antagonists of pome fruit tree diseases yielded two yellow epiphytic bacterial isolates morphologically similar to Pantoea agglomerans , but showing no biocontrol activity. Whole-cell MALDI-TOF mass spectrometry and analysis of 16S rRNA gene and gyrB sequences suggested the possibility of a novel species with a phylogenetic position in either the genus Pantoea or the genus Erwinia . Multi-locus sequence analysis (MLSA) placed the two strains in the genus Erwinia and supported their classification as a novel species. The strains showed general phenotypic characteristics typical of this genus and results of DNA-DNA hybridizations confirmed that they represent a single novel species. Both strains showed a DNA G+C content, as determined by HPLC, of 54.5 mol% and could be discriminated from phylogenetically related species of the genus Erwinia by their ability to utilize potassium gluconate, potassium 2-ketogluconate, maltose, melibiose and raffinose. Whole-genome sequencing of strain EM595 T revealed the presence of a chromosomal carotenoid biosynthesis gene cluster similar to those found in species of the genera Cronobacter and Pantoea that explains the pigmentation of the strain, which is atypical for the genus Erwinia . Additional strains belonging to the same species were recovered from different plant hosts in three different continents, revealing the cosmopolitan nature of this epiphyte. The name Erwinia gerundensis sp. nov. is proposed, with EM595 T ( = LMG 28990 T  = CCOS 903 T ) as the designated type strain.

Centrifugal partition chromatography: A preparative tool for isolation and purification of xylindein from Chlorociboria aeruginosa.

PubMed

Boonloed, Anukul; Weber, Genevieve L; Ramzy, Kelly M; Dias, Veronica R; Remcho, Vincent T

2016-12-23

A centrifugal partition chromatography (CPC) method was developed for the preparative-scale isolation and purification of xylindein from the wood-staining fungi, Chlorociboria aeruginosa. Xylindein, a blue-green pigment naturally secreted from the hyphae and fruiting bodies of the fungus, has great value in the decorative wood industry and textile coloration. Xylindein has great potential for use as a fluorescent labeling agent as well as in organic semiconductor applications. However, a primary limitation of xylindein is its poor solubility in most common HPLC solvents. Consequently, it is arduous to purify using preparative liquid chromatography or solid-phase extraction (SPE). Support-free, liquid-liquid chromatographic methods, including CPC, where solutes are separated based on their different distribution coefficients (K D ) between two immiscible solvent systems, are promising alternatives for the purification of the compound on a preparative scale. In this work, a new biphasic solvent system suitable for CPC separation of xylindein was developed. Various groups of solvents were assessed for their suitability as xylindein extractants. A new solvent system suitable for CPC separation of xylindein, composed of heptane/THF/MEK/acetonitrile/acetic acid/water, was developed. This solvent system yielded a K D value for xylindein of 1.54±0.04, as determined by HPLC (n=3). The compositions of the upper phase and lower phase of the solvent system were determined by Heteronuclear Single Quantum Correlation (HSQC) NMR and proton NMR. A CPC system, equipped with a fraction collector, was used for the isolation of xylindein from crude extracts. The xylindein fractions isolated by the CPC were then analyzed using HPLC and presented as a fractogram. Based on the CPC fractogram, the purified xylindein fractions were achieved after 30min CPC separation time, yielding 71% extraction efficiency. The developed CPC method allowed for isolation of this naturally sourced xylindein in amounts suitable for further study. Copyright © 2016 Elsevier B.V. All rights reserved.

Fingerprint analysis of polysaccharides from different Ganoderma by HPLC combined with chemometrics methods.

PubMed

Sun, Xiaomei; Wang, Haohao; Han, Xiaofeng; Chen, Shangwei; Zhu, Song; Dai, Jun

2014-12-19

A fingerprint analysis method has been developed for characterization and discrimination of polysaccharides from different Ganoderma by high performance liquid chromatography (HPLC) coupled with chemometrics means. The polysaccharides were extracted under ultrasonic-assisted condition, and then partly hydrolyzed with trifluoroacetic acid. Monosaccharides and oligosaccharides in the hydrolyzates were subjected to pre-column derivatization with 1-phenyl-3-methyl-5-pyrazolone and HPLC analysis, which will generate unique fingerprint information related to chemical composition and structure of polysaccharides. The peak data were imported to professional software in order to obtain standard fingerprint profiles and evaluate similarity of different samples. Meanwhile, the data were further processed by hierarchical cluster analysis and principal component analysis. Polysaccharides from different parts or species of Ganoderma or polysaccharides from the same parts of Ganoderma but from different geographical regions or different strains could be differentiated clearly. This fingerprint analysis method can be applied to identification and quality control of different Ganoderma and their products. Copyright © 2014 Elsevier Ltd. All rights reserved.

Effects of hemoglobin variants HbJ Bangkok, HbE, HbG Taipei, and HbH on analysis of glycated hemoglobin via ion-exchange high-performance liquid chromatography.

PubMed

Zhang, Xiu-Ming; Wen, Dong-Mei; Xu, Sheng-Nan; Suo, Ming-Huan; Chen, Ya-Qiong

2018-01-01

To explore the effects of HbJ Bangkok, HbE, HbG Taipei, and α-thalassemia HbH on the results of HbA1c assessment using ion-exchange high-performance liquid chromatography (IE-HPLC). We enrolled five patients in which the results of the IE-HPLC HbA1c assay were inconsistent with the average levels of FBG. We performed hemoglobin capillary (Hb) electrophoresis using whole-blood samples. We also sequenced the genes encoding Hb using dideoxy-mediated chain termination and analyzed HbA1c using borate affinity HPLC (BA-HPLC) and turbidimetric inhibition immunoassay (TINIA). Two patients had the HbJ Bangkok variant. Hb genotypes of these patients were β 41-42 /β J Bangkok and β N /β J Bangkok , and the content of HbJ Bangkok was 93.9% and 52.4%, respectively. The remaining three patients had the following: HbE (β N /β E Hb genotype, 23.6% HbE content), HbG Taipei (β N /β G Taipei Hb genotype, 39.4% HbG Taipei content), and α-thalassemia HbH (6.1% HbH content, 2.8% Hb Bart's content). In the patients with β-thalassemia and HbJ Bangkok variants, the presence of the variants interfered with the results of HbA1c analyses using IE-HPLC and TINIA; in the remaining four patients, there was interference with the results of HbA1c IE-HPLC but not with the TINIA assay. There was no interference with BA-HPLC HbA1c results. HbJ Bangkok, HbE, HbG Taipei Hb, and α-thalassemia HbH disease cause varying degrees of interference with the analysis of HbA1c using IE-HPLC. In these patients, we suggest using methods free from such interference for the analysis of HbA1c and other indicators to monitor blood glucose levels. © 2017 Wiley Periodicals, Inc.

Pigmented lesions of the oral mucosa: A cross-sectional study of 458 histopathological specimens.

PubMed

Tavares, Thalita Soares; Meirelles, Daniela Pereira; de Aguiar, Maria Cássia Ferreira; Caldeira, Patrícia Carlos

2018-06-26

To evaluate clinical, demographic, and histopathological characteristics of pigmented lesions of the oral mucosa. A cross-sectional study was conducted over a 64-year period. Information was collected from medical charts, and all archived histopathologic specimens with diagnoses of any pigmented lesion were retrieved. Statistical analysis was performed using SPSS software. A total of 34,127 archived specimens were reviewed, revealing 458 (1.34%) pigmented lesions, of which 230 were melanocytic and 228 non-melanocytic. Most patients were females (74.2%), white-skinned (49.1%), in the third and seventh decades of life (mean of 45 years). Most lesions were macular (59.8%), followed by plaques and nodules (4.8%), measuring 0-5 mm (41.9%). Cheek mucosa (21.0%), alveolar mucosa (16.6%), and gingiva (11.8%) were the most commonly affected sites. Amalgam tattoo was applied in 212 cases (46.3%), followed by melanotic macule (22.9%) and nevus (20.5%). Other diagnoses included racial pigmentation, exogenous pigmentation, melanotic neuroectodermal tumor of infancy, melanoma, melanocanthoma, smoker's melanosis, and heavy metal pigmentation. Pigmented lesions represent an uncommon diagnosis in oral pathology routines. The most frequent entities are amalgam tattoo, melanotic macule, and nevus. Patients are usually middle-aged women presenting a small, long-lasting, macular lesion on the cheek mucosa. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Phytoplankton Pigment Communities Can be Modeled Using Unique Relationships With Spectral Absorption Signatures in a Dynamic Coastal Environment

NASA Astrophysics Data System (ADS)

Catlett, D.; Siegel, D. A.

2018-01-01

Understanding the roles of phytoplankton community composition in the functioning of marine ecosystems and ocean biogeochemical cycles is important for many ocean science problems of societal relevance. Remote sensing currently offers the only feasible method for continuously assessing phytoplankton community structure on regional to global scales. However, methods are presently hindered by the limited spectral resolution of most satellite sensors and by uncertainties associated with deriving quantitative indices of phytoplankton community structure from phytoplankton pigment concentrations. Here we analyze a data set of concurrent phytoplankton pigment concentrations and phytoplankton absorption coefficient spectra from the Santa Barbara Channel, California, to develop novel optical oceanographic models for retrieving metrics of phytoplankton community composition. Cluster and Empirical Orthogonal Function analyses of phytoplankton pigment concentrations are used to define up to five phytoplankton pigment communities as a representation of phytoplankton functional types. Unique statistical relationships are found between phytoplankton pigment communities and absorption features isolated using spectral derivative analysis and are the basis of predictive models. Model performance is substantially better for phytoplankton pigment community indices compared with determinations of the contributions of individual pigments or taxa to chlorophyll a. These results highlight the application of data-driven chemotaxonomic approaches for developing and validating bio-optical algorithms and illustrate the potential and limitations for retrieving phytoplankton community composition from hyperspectral satellite ocean color observations.

The Association Between Melasma and Postinflammatory Hyperpigmentation in Acne Patients

PubMed Central

Adalatkhah, Hassan; Sadeghi Bazargani, Homayoun

2013-01-01

Background Although, melasma is most prevalent among Asian young women, and also darkly pigmented individuals are particularly prone to developing post inflammatory hyperpigmentation, to the best of our knowledge, there are rare or no studies about the association of melasma and Post inflammatory hyperpigmentation. Objectives The aim of this study was to investigate how likely is a melasma patient to developed post inflammatory hyperpigmentation when compared to patients with inflammatory acne lesions who do not have melasma. Patients and Methods This comparative study was conducted on 400 participants, 200 subjects involved with pigmented lesions of melasma and inflammatory acne lesions and200 involved only with inflammatory Acne lesions without melasma. Melasma, acne and post inflammatory hyper pigmentation, if existed, were assessed by a dermatologist, and pigmentation depth was assessed by wood's lamp. Multivariate logistic regression analysis suitable for study design was used to assess the association between melasma and post-acne pigmentation. Results We found out that 24.1% of patients without melasma had post-acne pigmentation compared to 66.8% in melasma group (P < 0.001). The likelihood of observing post-acne pigmentation was found to be nearly six times more in melasma patients versus those without melasma. Association existed after controlling for possible confounders such as melanin score and time length of self-reported sun exposure, and acne severity score. Conclusions Melasma appears to increase the likelihood of post-acne pigmentation. PMID:24349727

Development of space-stable thermal control coatings for use on large space vehicles

NASA Technical Reports Server (NTRS)

Gilligan, J. E.; Harada, Y.

1976-01-01

The potential of zinc orthotitanate as a pigment for spacecraft thermal control was demonstrated. The properties and performance of pigments prepared by solid state, coprecipitation, and mixed oxalate methods were compared. Environmental tests and subsequent spectral analysis were given primary emphasis.

Three-dimensional characterization of pigment dispersion in dried paint films using focused ion beam-scanning electron microscopy.

PubMed

Lin, Jui-Ching; Heeschen, William; Reffner, John; Hook, John

2012-04-01

The combination of integrated focused ion beam-scanning electron microscope (FIB-SEM) serial sectioning and imaging techniques with image analysis provided quantitative characterization of three-dimensional (3D) pigment dispersion in dried paint films. The focused ion beam in a FIB-SEM dual beam system enables great control in slicing paints, and the sectioning process can be synchronized with SEM imaging providing high quality serial cross-section images for 3D reconstruction. Application of Euclidean distance map and ultimate eroded points image analysis methods can provide quantitative characterization of 3D particle distribution. It is concluded that 3D measurement of binder distribution in paints is effective to characterize the order of pigment dispersion in dried paint films.

Analogs of Estrogen Metabolites as Probes of Estrogen-Induced Tumorigenesis

DTIC Science & Technology

1999-07-01

bromination reaction by reverse phase HPLC revealed a mixture of 4-bromoestradiol (5-10%), 2-bromoestradiol 28 (’-15%) and 2,4- dibromoestradiol 29...mixture. HPLC analysis of the reaction mixture revealed that the estradiol was completely consumed and 2,4-dibromoestradiol 29 was the major product...purification by HPLC .5 A solution of 30 in THF at -78’C was treated with various organolithium reagents and stirred for three hours after which the

Methods of Analysis by the U.S. Geological Survey Organic Geochemistry Research Group-Update and Additions to the Determination of Chloroacetanilide Herbicide Degradation Compounds in Water Using High-Performance Liquid Chromatography/Mass Spectrometry

USGS Publications Warehouse

Lee, E.A.; Kish, J.L.; Zimmerman, L.R.; Thurman, E.

2001-01-01

An analytical method using high-performance liquid chromatography/mass spectrometry (HPLC/MS) was developed by the U.S. Geological Survey in 1999 for the analysis of selected chloroacetanilide herbicide degradation compounds in water. These compounds were acetochlor ethane sulfonic acid (ESA), acetochlor oxanilic acid (OXA), alachlor ESA, alachlor OXA, metolachlor ESA, and metolachlor OXA. The HPLC/MS method was updated in 2000, and the method detection limits were modified accordingly. Four other degradation compounds also were added to the list of compounds that can be analyzed using HPLC/MS; these compounds were dimethenamid ESA, dimethenamid OXA, flufenacet ESA, and flufenacet OXA. Except for flufenacet OXA, good precision and accuracy were demonstrated for the updated HPLC/MS method in buffered reagent water, surface water, and ground water. The mean HPLC/MS recoveries of the degradation compounds from water samples spiked at 0.20 and 1.0 ?g/L (microgram per liter) ranged from 75 to 114 percent, with relative standard deviations of 15.8 percent or less for all compounds except flufenacet OXA, which had relative standard deviations ranging from 11.3 to 48.9 percent. Method detection levels (MDL's) using the updated HPLC/MS method varied from 0.009 to 0.045 ?g/L, with the flufenacet OXA MDL at 0.072 ?g/L. The updated HPLC/MS method is valuable for acquiring information about the fate and transport of the parent chloroacetanilide herbicides in water.

Screening and analysis of bioactive compounds with biofingerprinting chromatogram analysis of traditional Chinese medicines targeting DNA by microdialysis/HPLC.

PubMed

Su, Xingye; Kong, Liang; Li, Xin; Chen, Xueguo; Guo, Ming; Zou, Hanfa

2005-05-27

Biofingerprinting chromatogram analysis, which is defined as the comparison of fingerprinting chromatograms of the extract of traditional Chinese medicines (TCMs) before and after the interaction with biological systems (DNA, protein, cell, etc.), was proposed for screening and analysis of the multiple bioactive compounds in TCMs. A method of microdialysis sampling combined with high performance liquid chromatography (HPLC) was applied to the study of DNA-binding property for the extracts of TCMs. Seven compounds were found to bind to calf thymus DNA (ct-DNA) from the TCMs of Coptis chinensis Franch (Coptis), but only three ones from Phellodendron amurense Rupr. (Phellodendron) and none from Sophoraflavescens Ait. (Sophora) to bind to ct-DNA, respectively. Three of them were identified as berberine, palmatine and jatrorrhizine and their association constants (K) to ct-DNA were determined by microdialysis/HPLC. Competitive binding behaviors of them to ct-DNA were also investigated.

Analysis Study of Stevioside and Rebaudioside A from Stevia rebaudiana Bertoni by Normal Phase SPE and RP-HPLC

NASA Astrophysics Data System (ADS)

Martono, Y.; Rohman, A.; Riyanto, S.; Martono, S.

2018-04-01

Solid Phase Extraction (SPE) method using silica as sorbent for stevioside and rebaudiosida A analysis in Stevia rebaudiana Bertoni leaf have not been performed. The aim of this study is to develop SPE method using silica as sorbent for Reverse Phase-High Performance Liquid Chromatography (RP-HPLC) analysis of stevioside and rebaudiosida A in S. rebaudiana leaf. The results of this study indicate that the optimal conditions for normal phase SPE (silica) are conditioned with 3.0 mL of hexane. The sample loading volume is 0.1 mL. Cartridge is eluted with 1.0 mL acetonitrile: water (80: 20, v/v) to separate both analytes. The cartridge is washed with chloroform and water of 0.3 mL respectively. The developed SPE sample preparation method meets the accuracy and precision test and can be used for the analysis of stevioside and rebaudioside A by RP-HPLC.

A simple analytical procedure to replace HPLC for monitoring treatment concentrations of chloramine-T on fish culture facilities

USGS Publications Warehouse

Dawson, Verdel K.; Meinertz, Jeffery R.; Schmidt, Larry J.; Gingerich, William H.

2003-01-01

Concentrations of chloramine-T must be monitored during experimental treatments of fish when studying the effectiveness of the drug for controlling bacterial gill disease. A surrogate analytical method for analysis of chloramine-T to replace the existing high-performance liquid chromatography (HPLC) method is described. A surrogate method was needed because the existing HPLC method is expensive, requires a specialist to use, and is not generally available at fish hatcheries. Criteria for selection of a replacement method included ease of use, analysis time, cost, safety, sensitivity, accuracy, and precision. The most promising approach was to use the determination of chlorine concentrations as an indicator of chloramine-T. Of the currently available methods for analysis of chlorine, the DPD (N,N-diethyl-p-phenylenediamine) colorimetric method best fit the established criteria. The surrogate method was evaluated under a variety of water quality conditions. Regression analysis of all DPD colorimetric analyses with the HPLC values produced a linear model (Y=0.9602 X+0.1259) with an r2 value of 0.9960. The average accuracy (percent recovery) of the DPD method relative to the HPLC method for the combined set of water quality data was 101.5%. The surrogate method was also evaluated with chloramine-T solutions that contained various concentrations of fish feed or selected densities of rainbow trout. When samples were analyzed within 2 h, the results of the surrogate method were consistent with those of the HPLC method. When samples with high concentrations of organic material were allowed to age more than 2 h before being analyzed, the DPD method seemed to be susceptible to interference, possibly from the development of other chloramine compounds. However, even after aging samples 6 h, the accuracy of the surrogate DPD method relative to the HPLC method was within the range of 80–120%. Based on the data comparing the two methods, the U.S. Food and Drug Administration has concluded that the DPD colorimetric method is appropriate to use to measure chloramine-T in water during pivotal efficacy trials designed to support the approval of chloramine-T for use in fish culture.

Erroneous HbA1c results in a patient with elevated HbC and HbF.

PubMed

Adekanmbi, Joy; Higgins, Trefor; Rodriguez-Capote, Karina; Thomas, Dylan; Winterstein, Jeffrey; Dixon, Tara; Gifford, Jessica L; Krause, Richard; Venner, Allison A; Clarke, Gwen; Estey, Mathew P

2016-11-01

HbA1c is used in the diagnosis and monitoring of diabetes mellitus (DM). Interference from hemoglobin variants is a well-described phenomenon, particularly with HPLC-based methods. While immunoassays may generate more reliable HbA1c results in the presence of some variants, these methods are susceptible to negative interference from high concentrations of HbF. We report a case where an accurate HbA1c result could not be obtained by any available method due to the presence of a compound hemoglobinopathy. HbA1c was measured by HPLC, immunoassay, and capillary electrophoresis. Hemoglobinopathy investigation consisted of a CBC, hemoglobin fractionation by HPLC and electrophoresis, and molecular analysis. HbA1c analysis by HPLC and capillary electrophoresis gave no result. Analysis by immunoassay yielded HbA1c results of 5.9% (Siemens DCA 2000+) and 5.1% (Roche Integra), which were inconsistent with other markers of glycemic control. Hemoglobinopathy investigation showed HbC with the hereditary persistence of fetal hemoglobin-2 Ghana deletion. Reliable HbA1c results may be unobtainable in the presence of some hemoglobinopathies. HPLC and capillary electrophoresis alerted the laboratory to the presence of an unusual hemoglobinopathy. Immunoassays generated falsely low results without warning, which could lead to missed diagnoses and under treatment of patients with DM. Copyright © 2016 Elsevier B.V. All rights reserved.

Optical behaviour of copper phthalocyanine preparations for inkjet inks.

PubMed

Otáhalová, Lenka; Kaplanová, Marie; Gunde, Marta Klanjšek; Puchta, Miloš

2011-06-01

The present study investigates the preparation of the copper phthalocyanine pigment for inkjet printing inks. The pigment particle size distribution was measured with laser diffraction at different times of wet milling. Simultaneously, the absorbance spectra in a visible-near infrared spectral region of the corresponding diluted pigment dispersions were measured. At the beginning of the milling process, the particle size distribution is bimodal, showing the presence of aggregates and agglomerates. During the second hour of milling, the particle size distribution changes to unimodal due to the breaking of agglomerates, and the corresponding absorbance spectra change accordingly. Further milling diminishes the size of pigment aggregates up to the steady state value of around 130 nm, where also the absorbance in the corresponding spectra does not increase. A detailed analysis of intensity and position of the absorbance peak at 340 nm in dependence on the milling time and pigment concentration confirms the idea that an optical spectroscopy could be used for the assessment of optimal milling time required for the preparation of pigments with the maximum absorption ability.

Detailed Investigations on the Solid Cell Culture and Antimicrobial Activities of the Iranian Arnebia euchroma

PubMed Central

Haghbeen, K.; Pourmolaei, S.; Mareftjo, M. J.; Mousavi, A.; Akbari Noghabi, K.; Hosseini Shirazi, F.; Meshkat, A.

2011-01-01

In pursuit of strong shikalkin-producing cell lines, seeds of the Iranian Arnebia euchroma were collected from Dena altitudes in the central Zagross. Chemical analysis showed that the dried root of the plant contained about 8.5% (w/w) shikalkin pigment. The root explants of the young plantlets, obtained from the germinated seeds, were used for establishing callus. Then, parameters effective on proliferation and pigment production of the resulting calli were studied in detail. Accordingly, two modified media called mLS and mM9 were optimized for propagation and pigment production, respectively. Using these media, the biomass of the A.euchroma calli was increased to 600%, and the pigment production reached to a maximum of 16.3 mg per gram of the wet biomass in a period of a subculture (21 days). Parallel to these experiments, the antimicrobial activity of shikalkin pigment was examined on some fungi and Gram-positive and Gram-negative bacteria. Results indicated that the pigment was almost ineffective on fungi and Gram-negative bacteria, but it was meaningfully effective against Micrococcus luteus. PMID:21772789

Method optimization and quality assurance in speciation analysis using high performance liquid chromatography with detection by inductively coupled plasma mass spectrometry

NASA Astrophysics Data System (ADS)

Larsen, Erik H.

1998-02-01

Achievement of optimum selectivity, sensitivity and robustness in speciation analysis using high performance liquid chromatography (HPLC) with inductively coupled mass spectrometry (ICP-MS) detection requires that each instrumental component is selected and optimized with a view to the ideal operating characteristics of the entire hyphenated system. An isocratic HPLC system, which employs an aqueous mobile phase with organic buffer constituents, is well suited for introduction into the ICP-MS because of the stability of the detector response and high degree of analyte sensitivity attained. Anion and cation exchange HPLC systems, which meet these requirements, were used for the seperation of selenium and arsenic species in crude extracts of biological samples. Furthermore, the signal-to-noise ratios obtained for these incompletely ionized elements in the argon ICP were further enhanced by a factor of four by continously introducing carbon as methanol via the mobile phase into the ICP. Sources of error in the HPLC system (column overload), in the sample introduction system (memory by organic solvents) and in the ICP-MS (spectroscopic interferences) and their prevention are also discussed. The optimized anion and cation exchange HPLC-ICP-MS systems were used for arsenic speciation in contaminated ground water and in an in-house shrimp reference sample. For the purpose of verification, HPLC coupled with tandem mass spectrometry with electrospray ionization was additionally used for arsenic speciation in the shrimp sample. With this analytical technique the HPLC retention time in combination with mass analysis of the molecular ions and their collision-induced fragments provide almost conclusive evidence of the identity of the analyte species. The speciation methods are validated by establishing a mass balance of the analytes in each fraction of the extraction procedure, by recovery of spikes and by employing and comparing independent techniques. The urgent need for reference materials certified for elemental species is stressed.

Identification and characterization of the ergochrome gene cluster in the plant pathogenic fungus Claviceps purpurea.

PubMed

Neubauer, Lisa; Dopstadt, Julian; Humpf, Hans-Ulrich; Tudzynski, Paul

2016-01-01

Claviceps purpurea is a phytopathogenic fungus infecting a broad range of grasses including economically important cereal crop plants. The infection cycle ends with the formation of the typical purple-black pigmented sclerotia containing the toxic ergot alkaloids. Besides these ergot alkaloids little is known about the secondary metabolism of the fungus. Red anthraquinone derivatives and yellow xanthone dimers (ergochromes) have been isolated from sclerotia and described as ergot pigments, but the corresponding gene cluster has remained unknown. Fungal pigments gain increasing interest for example as environmentally friendly alternatives to existing dyes. Furthermore, several pigments show biological activities and may have some pharmaceutical value. This study identified the gene cluster responsible for the synthesis of the ergot pigments. Overexpression of the cluster-specific transcription factor led to activation of the gene cluster and to the production of several known ergot pigments. Knock out of the cluster key enzyme, a nonreducing polyketide synthase, clearly showed that this cluster is responsible for the production of red anthraquinones as well as yellow ergochromes. Furthermore, a tentative biosynthetic pathway for the ergot pigments is proposed. By changing the culture conditions, pigment production was activated in axenic culture so that high concentration of phosphate and low concentration of sucrose induced pigment syntheses. This is the first functional analysis of a secondary metabolite gene cluster in the ergot fungus besides that for the classical ergot alkaloids. We demonstrated that this gene cluster is responsible for the typical purple-black color of the ergot sclerotia and showed that the red and yellow ergot pigments are products of the same biosynthetic pathway. Activation of the gene cluster in axenic culture opened up new possibilities for biotechnological applications like the dye production or the development of new pharmaceuticals.

[HPLC fingerprint analysis of flavonoids of phyllanthi fructus from different habitats].

PubMed

Wang, Fei; Wang, Shuai; Meng, Xian-sheng; Bao, Yong-rui; Zhu, Ying-huan

2014-11-01

To establish the HPLC fingerprint of flavonoids of Phyllanthi Fructus from different habitats. HPLC method was adopted. The flavonoids composition of Phyllanthi Fructus from 10 different habitats was determined on an Agilent C, chromatographic column with 0. 5% formic acid water (A)-acetonitrile (B) as the mobile phase in gradient elution under the wavelength of 254 nm. The HPLC fingerprints of flavonoids composition of Phyllanthi Fructus were established to evaluate the qualitiy of them. The HPLC fingerprints of flavonoids composition of Phyllanthi Fructus from 10 different habitats were established. 18 common peaks were found and the similarities of them were more than 0. 90 except the ones from Guangxi and Guangdong. The method is simple, accurate and repeatable. It can be used for research and quality control of the effective components in Phyllanthi Fructus.

Analysis and identification of astaxanthin and its carotenoid precursors from Xanthophyllomyces dendrorhous by high-performance liquid chromatography.

PubMed

Lu, Mingbo; Zhang, Yang'e; Zhao, Chunfang; Zhou, Pengpeng; Yu, Longjiang

2010-01-01

This study presents an HPLC method for simultaneous analysis of astaxanthin and its carotenoid precursors from Xanthophyllomyces dendrorhous. The HPLC method is accomplished by employing a C18 column and the mobile phase methanol/water/acetonitrile/ dichloromethane (70:4:13:13, v/v/v/v). Astaxanthin is quantified by detection at 480 nm. The carotenoid precursors are identified by LC-APCI-MS and UV-vis absorption spectra. Peaks showed in the HPLC chromatogram are identified as carotenoids in the monocyclic biosynthetic pathway or their derivatives. In the monocyclic carotenoid pathway, 3,3'-dihydroxy-beta,psi-carotene-4,4'-dione (DCD) is produced through gamma-carotene and torulene.

Dipeptide Sequence Determination: Analyzing Phenylthiohydantoin Amino Acids by HPLC

NASA Astrophysics Data System (ADS)

Barton, Janice S.; Tang, Chung-Fei; Reed, Steven S.

2000-02-01

Amino acid composition and sequence determination, important techniques for characterizing peptides and proteins, are essential for predicting conformation and studying sequence alignment. This experiment presents improved, fundamental methods of sequence analysis for an upper-division biochemistry laboratory. Working in pairs, students use the Edman reagent to prepare phenylthiohydantoin derivatives of amino acids for determination of the sequence of an unknown dipeptide. With a single HPLC technique, students identify both the N-terminal amino acid and the composition of the dipeptide. This method yields good precision of retention times and allows use of a broad range of amino acids as components of the dipeptide. Students learn fundamental principles and techniques of sequence analysis and HPLC.

HPLC analysis of functionalized poly(amidoamine) dendrimers and the interaction between a folate-dendrimer conjugate and folate binding protein.

PubMed

Shi, Xiangyang; Bi, Xiangdong; Ganser, T Rose; Hong, Seungpyo; Myc, Lukasz A; Desai, Ankur; Holl, Mark M Banaszak; Baker, James R

2006-07-01

Poly(amidoamine) (PAMAM) dendrimers of different generations with carboxyl, acetyl, and hydroxyl terminal groups and a folic acid (FA)-dendrimer conjugate were separated and analyzed using reverse-phase high performance liquid chromatography (HPLC). Analysis of both the individual PAMAM derivatives and the separation of mixed generations can be achieved using a linear gradient 0-50% acetonitrile (ACN) (balance water) within 40 min. We also show that PAMAMs with defined acetylation and carboxylation degrees can be analyzed using HPLC. Furthermore, a generation 5 dendrimer-FA conjugate (G5.75Ac-FA4; Ac denotes acetyl) was analyzed and its specific binding with a bovine folic acid binding protein (FBP) was monitored. The HPLC and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results indicate the formation of three complexes after the binding of G5.75Ac-FA4 with FBP. Dendrimers with FA moieties show much higher specific binding capability with FBP than those without FA moieties. Findings from this study indicate that HPLC is an effective technique not only for characterization and separation of functionalized PAMAM dendrimers and conjugates but also for investigation of the interaction between dendrimers and biomolecules.

Regio- and stereospecific analysis of glycerolipids.

PubMed

Kuksis, Arnis; Itabashi, Yutaka

2005-06-01

In recent years researchers have recognized the potential value of comprehensive lipid profiling (lipidomics), which was invented and promoted by lipidologists who recognized the many valuable applications that grew out of the fields of DNA profiling (genomics) and protein profiling (proteonomics). Through lipid class-selective intrasource ionization and subsequent analysis of two-dimensional cross-peak intensities, the chemical identity and mass composition of individual molecular species of most lipid classes can now be determined in a chloroform extract. There remains, however, the necessity to distinguish the enantiomers and isobaric regioisomers resulting from enzymatic and chemical reactions, which conventional high performance liquid chromatography/mass spectrometry (HPLC/MS) has been slow to accommodate, and tandem MS unable to provide. While reversed-phase HPLC can separate regioisomers, normal-phase HPLC can resolve diastereomers, and chiral-phase HPLC can effect dramatic resolution of enantiomers, the full potential of the combined systems has seldom been exploited. The present chapter calls attention to both recent and earlier combinations of these methodologies with mass spectrometry, which allows the HPLC/ESI (electrospray ionization)-MS/MS separation and identification of enantiomeric diacylglycerols, triacylglycerols, and glycerophospholipids as well as their isobaric regioisomers. These developments permit further expansion of lipid profiling (lipidomics) and better understanding of lipid metabolism.

X-ray fluorescence and imaging analyses of paintings by the Brazilian artist Oscar Pereira Da Silva

NASA Astrophysics Data System (ADS)

Campos, P. H. O. V.; Kajiya, E. A. M.; Rizzutto, M. A.; Neiva, A. C.; Pinto, H. P. F.; Almeida, P. A. D.

2014-02-01

Non-destructive analyses, such as EDXRF (Energy-Dispersive X-Ray Fluorescence) spectroscopy, and imaging were used to characterize easel paintings. The analyzed objects are from the collection of the Pinacoteca do Estado de São Paulo. EDXRF results allowed us to identify the chemical elements present in the pigments, showing the use of many Fe-based pigments, modern pigments, such as cobalt blue and cadmium yellow, as well as white pigments containing lead and zinc used by the artist in different layers. Imaging analysis was useful to identify the state of conservation, the localization of old and new restorations and also to detect and unveil the underlying drawings revealing the artist's creative processes.

Bioavailability of plant pigment phytochemicals in Angelica keiskei in older adults: A pilot absorption kinetic study

PubMed Central

Correa, Camila R; Chen, C-Y. Oliver; Aldini, Giancarlo; Rasmussen, Helen; Ronchi, Carlos F; Berchieri-Ronchi, Carolina; Cho, Soo-Muk; Blumberg, Jeffrey B

2014-01-01

BACKGROUND/OBJECTIVES Angelica keiskei is a green leafy vegetable rich in plant pigment phytochemicals such as flavonoids and carotenoids. This study examined bioavailability of flavonoids and carotenoids in Angelica keiskei and the alteration of the antioxidant performance in vivo. SUBJECTS AND MATERIALS Absorption kinetics of phytochemicals in Angelica keiskei were determined in healthy older adults (> 60 y, n = 5) and subjects with metabolic syndrome (n = 5). Subjects consumed 5 g dry Angelica keiskei powder encapsulated in gelatin capsules with a low flavonoid and carotenoid liquid meal. Plasma samples were collected at baseline, 0.5, 1, 2, 3, 4, 5, 6, 7, and 8 h. Samples were analyzed for flavonoids and carotenoids using HPLC systems with electrochemical and UV detection, respectively, and for total antioxidant performance by fluorometry. RESULTS After ingestion of Angelica keiskei increases in plasma quercetin concentrations were observed at 1-3 and 6-8 hr in the healthy group and at all time points in the metabolic syndrome group compared to baseline (P < 0.05). Plasma lutein concentrations were significantly elevated in both the healthy and metabolic syndrome groups at 8 hr (P < 0.05). Significant increases in total antioxidant performance were also observed in both the healthy and the metabolic syndrome groups compared to baseline (P < 0.05). CONCLUSIONS Findings of this study clearly demonstrate the bioavailability of phytonutrients of Angelica keiskei and their ability to increase antioxidant status in humans. PMID:25324936

Detection of pigments of halophilic endoliths from gypsum: Raman portable instrument and European Space Agency's prototype analysis.

PubMed

Culka, Adam; Osterrothová, Kateřina; Hutchinson, Ian; Ingley, Richard; McHugh, Melissa; Oren, Aharon; Edwards, Howell G M; Jehlička, Jan

2014-12-13

A prototype instrument, under development at the University of Leicester, for the future European Space Agency (ESA) ExoMars mission, was used for the analysis of microbial pigments within a stratified gypsum crust from a hypersaline saltern evaporation pond at Eilat (Israel). Additionally, the same samples were analysed using a miniaturized Raman spectrometer, featuring the same 532 nm excitation. The differences in the position of the specific bands, attributed to carotenoid pigments from different coloured layers, were minor when analysed by the ESA prototype instrument; therefore, making it difficult to distinguish among the different pigments. The portable Delta Nu Advantage instrument allowed for the discrimination of microbial carotenoids from the orange/green and purple layers. The purpose of this study was to complement previous laboratory results with new data and experience with portable or handheld Raman systems, even with a dedicated prototype Raman system for the exploration of Mars. The latter is equipped with an excitation wavelength falling within the carotenoid polyene resonance region. The ESA prototype Raman instrument detected the carotenoid pigments (biomarkers) with ease, although further detailed distinctions among them were not achieved. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

In Silico Analysis of Gene Expression Network Components Underlying Pigmentation Phenotypes in the Python Identified Evolutionarily Conserved Clusters of Transcription Factor Binding Sites

PubMed Central

2016-01-01

Color variation provides the opportunity to investigate the genetic basis of evolution and selection. Reptiles are less studied than mammals. Comparative genomics approaches allow for knowledge gained in one species to be leveraged for use in another species. We describe a comparative vertebrate analysis of conserved regulatory modules in pythons aimed at assessing bioinformatics evidence that transcription factors important in mammalian pigmentation phenotypes may also be important in python pigmentation phenotypes. We identified 23 python orthologs of mammalian genes associated with variation in coat color phenotypes for which we assessed the extent of pairwise protein sequence identity between pythons and mouse, dog, horse, cow, chicken, anole lizard, and garter snake. We next identified a set of melanocyte/pigment associated transcription factors (CREB, FOXD3, LEF-1, MITF, POU3F2, and USF-1) that exhibit relatively conserved sequence similarity within their DNA binding regions across species based on orthologous alignments across multiple species. Finally, we identified 27 evolutionarily conserved clusters of transcription factor binding sites within ~200-nucleotide intervals of the 1500-nucleotide upstream regions of AIM1, DCT, MC1R, MITF, MLANA, OA1, PMEL, RAB27A, and TYR from Python bivittatus. Our results provide insight into pigment phenotypes in pythons. PMID:27698666

In Silico Analysis of Gene Expression Network Components Underlying Pigmentation Phenotypes in the Python Identified Evolutionarily Conserved Clusters of Transcription Factor Binding Sites.

PubMed

Irizarry, Kristopher J L; Bryden, Randall L

2016-01-01

Color variation provides the opportunity to investigate the genetic basis of evolution and selection. Reptiles are less studied than mammals. Comparative genomics approaches allow for knowledge gained in one species to be leveraged for use in another species. We describe a comparative vertebrate analysis of conserved regulatory modules in pythons aimed at assessing bioinformatics evidence that transcription factors important in mammalian pigmentation phenotypes may also be important in python pigmentation phenotypes. We identified 23 python orthologs of mammalian genes associated with variation in coat color phenotypes for which we assessed the extent of pairwise protein sequence identity between pythons and mouse, dog, horse, cow, chicken, anole lizard, and garter snake. We next identified a set of melanocyte/pigment associated transcription factors (CREB, FOXD3, LEF-1, MITF, POU3F2, and USF-1) that exhibit relatively conserved sequence similarity within their DNA binding regions across species based on orthologous alignments across multiple species. Finally, we identified 27 evolutionarily conserved clusters of transcription factor binding sites within ~200-nucleotide intervals of the 1500-nucleotide upstream regions of AIM1, DCT, MC1R, MITF, MLANA, OA1, PMEL, RAB27A, and TYR from Python bivittatus . Our results provide insight into pigment phenotypes in pythons.

Clinical and dermoscopic clues to differentiate pigmented nail bands: an International Dermoscopy Society study.

PubMed

Benati, E; Ribero, S; Longo, C; Piana, S; Puig, S; Carrera, C; Cicero, F; Kittler, H; Deinlein, T; Zalaudek, I; Stolz, W; Scope, A; Pellacani, G; Moscarella, E; Piraccini, B M; Starace, M; Argenziano, G

2017-04-01

Longitudinal melanonychia might be difficult to differentiate and the use of dermoscopy can be useful for the preoperative evaluation and management decision. The aim of our study was to investigate clinical and dermoscopic criteria of acquired longitudinal melanonychia in adults to identify the best predictors of melanoma using a multivariate analysis and to explore eventual new dermoscopic criteria for nail melanoma diagnosis. In this retrospective observational study, 82 histopathologically diagnosed, acquired nail pigmented bands were collected and examined. All variables were included in the analysis and examined as possible predictors of nail melanoma. Both univariate and multivariable analyses have been performed. Among 82 cases, 25 were diagnosed as nail melanoma and 57 as benign lesions (including 32 melanocytic nevi and 25 benign melanocytic hyperplasia). Melanoma cases were significantly associated with a width of the pigmented band higher than 2/3 of the nail plate, grey and black colours, irregularly pigmented lines, Hutchinson and micro-Hutchinson signs, and nail dystrophy. Granular pigmentation, a newly defined dermoscopic criterion, was found in 40% of melanomas and only in 3.51% of benign lesions. Dermoscopic examination of longitudinal melanonychia provides useful information that could help clinicians to improve melanoma recognition. © 2016 European Academy of Dermatology and Venereology.

Remote-Sensing Estimation of Phytoplankton Size Classes From GOCI Satellite Measurements in Bohai Sea and Yellow Sea

NASA Astrophysics Data System (ADS)

Sun, Deyong; Huan, Yu; Qiu, Zhongfeng; Hu, Chuanmin; Wang, Shengqiang; He, Yijun

2017-10-01

Phytoplankton size class (PSC), a measure of different phytoplankton functional and structural groups, is a key parameter to the understanding of many marine ecological and biogeochemical processes. In turbid waters where optical properties may be influenced by terrigenous discharge and nonphytoplankton water constituents, remote estimation of PSC is still a challenging task. Here based on measurements of phytoplankton diagnostic pigments, total chlorophyll a, and spectral reflectance in turbid waters of Bohai Sea and Yellow Sea during summer 2015, a customized model is developed and validated to estimate PSC in the two semienclosed seas. Five diagnostic pigments determined through high-performance liquid chromatography (HPLC) measurements are first used to produce weighting factors to model phytoplankton biomass (using total chlorophyll a as a surrogate) with relatively high accuracies. Then, a common method used to calculate contributions of microphytoplankton, nanophytoplankton, and picophytoplankton to the phytoplankton assemblage (i.e., Fm, Fn, and Fp) is customized using local HPLC and other data. Exponential functions are tuned to model the size-specific chlorophyll a concentrations (Cm, Cn, and Cp for microphytoplankton, nanophytoplankton, and picophytoplankton, respectively) with remote-sensing reflectance (Rrs) and total chlorophyll a as the model inputs. Such a PSC model shows two improvements over previous models: (1) a practical strategy (i.e., model Cp and Cn first, and then derive Cm as C-Cp-Cn) with an optimized spectral band (680 nm) for Rrs as the model input; (2) local parameterization, including a local chlorophyll a algorithm. The performance of the PSC model is validated using in situ data that were not used in the model development. Application of the PSC model to GOCI (Geostationary Ocean Color Imager) data leads to spatial and temporal distribution patterns of phytoplankton size classes (PSCs) that are consistent with results reported from field measurements by other researchers. While the applicability of the PSC model together with its parameterization to other optically complex regions and to other seasons is unknown, the findings of this study suggest that the approach to develop such a model may be extendable to other cases as long as local data are used to select the optimal band and to determine the model coefficients.

The Impact of Quantitative Data Provided by a Multi-spectral Digital Skin Lesion Analysis Device on Dermatologists'Decisions to Biopsy Pigmented Lesions.

PubMed

Farberg, Aaron S; Winkelmann, Richard R; Tucker, Natalie; White, Richard; Rigel, Darrell S

2017-09-01

BACKGROUND: Early diagnosis of melanoma is critical to survival. New technologies, such as a multi-spectral digital skin lesion analysis (MSDSLA) device [MelaFind, STRATA Skin Sciences, Horsham, Pennsylvania] may be useful to enhance clinician evaluation of concerning pigmented skin lesions. Previous studies evaluated the effect of only the binary output. OBJECTIVE: The objective of this study was to determine how decisions dermatologists make regarding pigmented lesion biopsies are impacted by providing both the underlying classifier score (CS) and associated probability risk provided by multi-spectral digital skin lesion analysis. This outcome was also compared against the improvement reported with the provision of only the binary output. METHODS: Dermatologists attending an educational conference evaluated 50 pigmented lesions (25 melanomas and 25 benign lesions). Participants were asked if they would biopsy the lesion based on clinical images, and were asked this question again after being shown multi-spectral digital skin lesion analysis data that included the probability graphs and classifier score. RESULTS: Data were analyzed from a total of 160 United States board-certified dermatologists. Biopsy sensitivity for melanoma improved from 76 percent following clinical evaluation to 92 percent after quantitative multi-spectral digital skin lesion analysis information was provided ( p <0.0001). Specificity improved from 52 percent to 79 percent ( p <0.0001). The positive predictive value increased from 61 percent to 81 percent ( p <0.01) when the quantitative data were provided. Negative predictive value also increased (68% vs. 91%, p<0.01), and overall biopsy accuracy was greater with multi-spectral digital skin lesion analysis (64% vs. 86%, p <0.001). Interrater reliability improved (intraclass correlation 0.466 before, 0.559 after). CONCLUSION: Incorporating the classifier score and probability data into physician evaluation of pigmented lesions led to both increased sensitivity and specificity, thereby resulting in more accurate biopsy decisions.

Ion chromatography for the precise analysis of chloride and sodium in sweat for the diagnosis of cystic fibrosis.

PubMed

Doorn, J; Storteboom, T T R; Mulder, A M; de Jong, W H A; Rottier, B L; Kema, I P

2015-07-01

Measurement of chloride in sweat is an essential part of the diagnostic algorithm for cystic fibrosis. The lack in sensitivity and reproducibility of current methods led us to develop an ion chromatography/high-performance liquid chromatography (IC/HPLC) method, suitable for the analysis of both chloride and sodium in small volumes of sweat. Precision, linearity and limit of detection of an in-house developed IC/HPLC method were established. Method comparison between the newly developed IC/HPLC method and the traditional Chlorocounter was performed, and trueness was determined using Passing Bablok method comparison with external quality assurance material (Royal College of Pathologists of Australasia). Precision and linearity fulfill criteria as established by UK guidelines are comparable with inductively coupled plasma-mass spectrometry methods. Passing Bablok analysis demonstrated excellent correlation between IC/HPLC measurements and external quality assessment target values, for both chloride and sodium. With a limit of quantitation of 0.95 mmol/L, our method is suitable for the analysis of small amounts of sweat and can thus be used in combination with the Macroduct collection system. Although a chromatographic application results in a somewhat more expensive test compared to a Chlorocounter test, more accurate measurements are achieved. In addition, simultaneous measurements of sodium concentrations will result in better detection of false positives, less test repeating and thus faster and more accurate and effective diagnosis. The described IC/HPLC method, therefore, provides a precise, relatively cheap and easy-to-handle application for the analysis of both chloride and sodium in sweat. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.