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Sample records for rapid colorimetric quantification

  1. Rapid quantification of microalgal lipids in aqueous medium by a simple colorimetric method.

    PubMed

    Mishra, Sanjiv K; Suh, William I; Farooq, Wasif; Moon, Myounghoon; Shrivastav, Anupama; Park, Min S; Yang, Ji-Won

    2014-03-01

    Identification of novel microalgal strains with high lipid productivity is one of the most important research topics in renewable biofuel research. However, the major bottleneck in the strain screening process is that currently known methods for the estimation of microalgal lipid are laborious and time-consuming. The present study successfully employed sulpho-phospho-vanillin (SPV) colorimetric method for direct quantitative measurement of lipids within liquid microalgal culture. The SPV reacts with lipids to produce a distinct pink color, and its intensity can be quantified using spectrophotometric methods by measuring absorbance at 530nm. This method was employed for a rapid quantification of intracellular lipid contents within Chlorella sp., Monoraphidium sp., Ettlia sp. and Nannochloropsis sp., all of which were found to have lipid contents ranging in between 10% and 30%. Subsequent analysis of the biomass using gas chromatography confirmed that our protocol is highly accurate (R(2)=0.99). PMID:24463407

  2. A quick colorimetric method for total lipid quantification in microalgae.

    PubMed

    Byreddy, Avinesh R; Gupta, Adarsha; Barrow, Colin J; Puri, Munish

    2016-06-01

    Discovering microalgae with high lipid productivity are among the key milestones for achieving sustainable biodiesel production. Current methods of lipid quantification are time intensive and costly. A rapid colorimetric method based on sulfo-phospho-vanillin (SPV) reaction was developed for the quantification of microbial lipids to facilitate screening for lipid producing microalgae. This method was successfully tested on marine thraustochytrid strains and vegetable oils. The colorimetric method results correlated well with gravimetric method estimates. The new method was less time consuming than gravimetric analysis and is quantitative for lipid determination, even in the presence of carbohydrates, proteins and glycerol. PMID:27050419

  3. Rapid identification of bacteria with a disposable colorimetric sensing array.

    PubMed

    Carey, James R; Suslick, Kenneth S; Hulkower, Keren I; Imlay, James A; Imlay, Karin R C; Ingison, Crystal K; Ponder, Jennifer B; Sen, Avijit; Wittrig, Aaron E

    2011-05-18

    Rapid identification of both species and even specific strains of human pathogenic bacteria grown on standard agar has been achieved from the volatiles they produce using a disposable colorimetric sensor array in a Petri dish imaged with an inexpensive scanner. All 10 strains of bacteria tested, including Enterococcus faecalis and Staphylococcus aureus and their antibiotic-resistant forms, were identified with 98.8% accuracy within 10 h, a clinically important time frame. Furthermore, the colorimetric sensor arrays also proved useful as a simple research tool for the study of bacterial metabolism and as an easy method for the optimization of bacterial production of fine chemicals or other fermentation processes. PMID:21524080

  4. Rapid Quantification of Trimethylamine.

    PubMed

    Li, Zheng; Li, Hao; LaGasse, Maria K; Suslick, Kenneth S

    2016-06-01

    Sensitive detection of trimethylamine both in aqueous and gaseous phases has been accomplished using an inexpensive colorimetric sensor array. Distinctive color change patterns provide facile discrimination over a wide range of concentrations for trimethylamine with >99% accuracy of classification. Calculated limits of detection are well below the diagnostically significant concentration for trimethylaminuria (fish malodor syndrome). The sensor array shows good reversibility after multiple uses and is able to cleanly discriminate trimethylamine from similar amine odorants. Portable sensing of trimethylamine vapors at ppb concentrations is described using a cell phone camera or a hand-held optoelectronic nose. Application of the sensor array in detecting mouth and skin odor as a potential tool for portable diagnosis of trimethylaminuria is also illustrated. PMID:27220015

  5. Colorimetric Quantification and in Situ Detection of Collagen

    ERIC Educational Resources Information Center

    Esteban, Francisco J.; del Moral, Maria L.; Sanchez-Lopez, Ana M.; Blanco, Santos; Jimenez, Ana; Hernandez, Raquel; Pedrosa, Juan A.; Peinado, Maria A.

    2005-01-01

    A simple multidisciplinary and inexpensive laboratory exercise is proposed, in which the undergraduate student may correlate biochemical and anatomical findings. The entire practical session can be completed in one 2.5-3 hour laboratory period, and consists of the quantification of collagen and total protein content from tissue sections--without…

  6. Quantification of Wheat Grain Arabinoxylans Using a Phloroglucinol Colorimetric Assay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Arabinoxylans (AX) play a critical role in end-use quality and nutrition of wheat (Triticum aestivum L.). An efficient, accurate method of AX quantification is desirable as AX plays an important role in processing, end use quality and human health. The objective of this work was to evaluate a stand...

  7. Direct Quantification of Carotenoids in Low Fat Baby Foods Via Laser Photoacoustics and Colorimetric Index *

    NASA Astrophysics Data System (ADS)

    Dóka, O.; Ajtony, Zs.; Bicanic, D.; Valinger, D.; Végvári, Gy.

    2014-12-01

    Carotenoids are important antioxidants found in various foods including those for nutrition of infants. In this investigation, the total carotenoid content (TCC) of nine different commercially available baby foods was quantified using colorimetric index * obtained via reflectance colorimetry (RC) and by laser photoacoustic spectroscopy (LPAS) at 473 nm. The latter requires a minimum of sample preparation and only a one time calibration step which enables practically direct quantification of TCC. Results were verified versus UV-Vis spectrophotometry (SP) as the reference technique. It was shown that RC and LPAS (at 473 nm) provide satisfactory results for *, = 0.9925 and = 0.9972, respectively. Other color indices do not show a correlation with TCC. When determining the TCC in baby foods containing tomatoes, it is necessary to select a different analytical wavelength to compensate for the effect of lycopene's presence in the test samples.

  8. Rapid colorimetric sensing of tetracycline antibiotics with in situ growth of gold nanoparticles.

    PubMed

    Shen, Li; Chen, Jing; Li, Na; He, Pingli; Li, Zhen

    2014-08-11

    A colorimetric assay utilizing the formation of gold nanoparticles was developed to detect tetracycline antibiotics in fluidic samples. Tetracycline antibiotics showed the capability of directly reducing aurate salts into atomic gold which form gold nanoparticles spontaneously under proper conditions. The resulted gold nanoparticles showed characteristic plasmon absorbance at 526 nm, which can be visualized by naked eyes or with a spectrophotometer. UV-vis absorbance of the resulted gold nanoparticles is correlated directly with the concentrations of tetracycline antibiotics in the solution, allowing for quantitative colorimetric detection of tetracycline antibiotics. Reaction conditions, such as pH, temperature, reaction time, and ionic strength were optimized. Sensitivity of the colorimetric assay can be enhanced by the addition of gold nanoparticle seeds, a LOD as low as 20 ng mL(-1) can be achieved with the help of seed particles. The colorimetric assay showed minimum interference from ethanol, methanol, urea, glucose, and other antibiotics such as sulfonamides, amino glycosides etc. Validity of the method was also evaluated on urine samples spiked with tetracycline antibiotics. The method provides a broad spectrum detection method for rapid and sensitive detection of reductive substances such as tetracycline antibiotics in liquid and biological samples. PMID:25066722

  9. A Rapid Colorimetric Sensor of Clenbuterol Based on Cysteamine-Modified Gold Nanoparticles.

    PubMed

    Kang, Jingyan; Zhang, Yujie; Li, Xing; Miao, Lijing; Wu, Aiguo

    2016-01-13

    Demonstrated was a simple visual and rapid colorimetric sensor for detection of clenbuterol (CLB) based on gold nanoparticles (AuNPs) modified with cysteamine (CA) and characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), UV-vis. The solution color from red to blue gray with increasing clenbuterol concentration resulted from the aggregation of AuNPs. The detection limit of clenbuterol is 50 nM by naked eyes. The selectivity of CA-AuNPs detection system for clenbuterol is excellent compared with other interferents in food. This sensor has been successfully applied to detect clenbuterol in real blood sample. PMID:26673452

  10. A Simple Paper-Based Colorimetric Device for Rapid Mercury(II) Assay.

    PubMed

    Chen, Weiwei; Fang, Xueen; Li, Hua; Cao, Hongmei; Kong, Jilie

    2016-01-01

    Contamination of the environment by mercury(II) ions (Hg(2+)) poses a serious threat to human health and ecosystems. Up to now, many reported Hg(2+) sensors require complex procedures, long measurement times and sophisticated instrumentation. We have developed a simple, rapid, low cost and naked-eye quantitative method for Hg(2+) environmental analysis using a paper-based colorimetric device (PCD). The sample solution to which platinum nanoparticles (PtNPs) have been added is dispensed to the detection zone on the PCD, where the 3,3,5,5-tetramethylbenzidine (TMB) substrate has been pre-loaded. The PtNPs effect a rapid oxidization of TMB, inducing blue colorization on the PCD. However, Hg(2+) in the solution rapidly interact with the PtNPs, suppressing the oxidation capacity and hence causing a decrease in blue intensity, which can be observed directly by the naked eye. Moreover, Hg(2+) at concentrations as low as 0.01 uM, can be successfully monitored using a fiber optic device, which gives a digital readout proportional to the intensity of the blue color change. This paper-based colorimetric device (PCD) shows great potential for field measurement of Hg(2+). PMID:27554633

  11. A Simple Paper-Based Colorimetric Device for Rapid Mercury(II) Assay

    PubMed Central

    Chen, Weiwei; Fang, Xueen; Li, Hua; Cao, Hongmei; Kong, Jilie

    2016-01-01

    Contamination of the environment by mercury(II) ions (Hg2+) poses a serious threat to human health and ecosystems. Up to now, many reported Hg2+ sensors require complex procedures, long measurement times and sophisticated instrumentation. We have developed a simple, rapid, low cost and naked-eye quantitative method for Hg2+ environmental analysis using a paper-based colorimetric device (PCD). The sample solution to which platinum nanoparticles (PtNPs) have been added is dispensed to the detection zone on the PCD, where the 3,3,5,5-tetramethylbenzidine (TMB) substrate has been pre-loaded. The PtNPs effect a rapid oxidization of TMB, inducing blue colorization on the PCD. However, Hg2+ in the solution rapidly interact with the PtNPs, suppressing the oxidation capacity and hence causing a decrease in blue intensity, which can be observed directly by the naked eye. Moreover, Hg2+ at concentrations as low as 0.01 uM, can be successfully monitored using a fiber optic device, which gives a digital readout proportional to the intensity of the blue color change. This paper-based colorimetric device (PCD) shows great potential for field measurement of Hg2+. PMID:27554633

  12. A Rapid In Situ Colorimetric Assay for Cobalt Detection by the Naked Eye.

    PubMed

    Kang, Sung-Min; Jang, Sung-Chan; Kim, Gi Yong; Lee, Chang-Soo; Huh, Yun Suk; Roh, Changhyun

    2016-01-01

    A simple, rapid, and convenient colorimetric chemosensor of a specific target toward the end user is still required for on-site detection and real-time monitoring applications. In this study, we developed a rapid in situ colorimetric assay for cobalt detection using the naked eye. Interestingly, a yellow to light orange visual color transition was observed within 3 s when a Chrysoidine G (CG) chemosensor was exposed to cobalt. Surprisingly, the CG chemosensor had great selectivity toward cobalt without any interference of other metal ions. Under optimized conditions, a lower detection limit of 0.1 ppm via a spectrophotometer and a visual detection limit of 2 ppm with a linear range from 0.4 to 1 ppm (R² = 0.97) were determined. Moreover, the CG chemosensor is reversible and maintains its functionality after treatment with chelating agents. In conclusion, we show the superior capabilities of the CG chemosensor, which has the potential to provide extremely facile handling, high sensitivity, and a fast response time for applications of on-site detection to real-time cobalt monitoring for the general public. PMID:27144568

  13. A Rapid In Situ Colorimetric Assay for Cobalt Detection by the Naked Eye

    PubMed Central

    Kang, Sung-Min; Jang, Sung-Chan; Kim, Gi Yong; Lee, Chang-Soo; Huh, Yun Suk; Roh, Changhyun

    2016-01-01

    A simple, rapid, and convenient colorimetric chemosensor of a specific target toward the end user is still required for on-site detection and real-time monitoring applications. In this study, we developed a rapid in situ colorimetric assay for cobalt detection using the naked eye. Interestingly, a yellow to light orange visual color transition was observed within 3 s when a Chrysoidine G (CG) chemosensor was exposed to cobalt. Surprisingly, the CG chemosensor had great selectivity toward cobalt without any interference of other metal ions. Under optimized conditions, a lower detection limit of 0.1 ppm via a spectrophotometer and a visual detection limit of 2 ppm with a linear range from 0.4 to 1 ppm (R2 = 0.97) were determined. Moreover, the CG chemosensor is reversible and maintains its functionality after treatment with chelating agents. In conclusion, we show the superior capabilities of the CG chemosensor, which has the potential to provide extremely facile handling, high sensitivity, and a fast response time for applications of on-site detection to real-time cobalt monitoring for the general public. PMID:27144568

  14. Colorimetric Quantification of Glucose and Cholesterol in Human Blood Using a Nanocomposite Entrapping Magnetic Nanoparticles and Oxidases.

    PubMed

    Kim, Moon Il; Cho, Daeyeon; Park, Hyun Gyu

    2015-10-01

    In this study, a microscale well-plate colorimetric assay for the multiplexed detection of glucose and cholesterol in clinical human blood samples has been developed. This system utilized one-pot nanocomposite entrapping Fe3O4 magnetic nanoparticles (MNPs) as peroxidase mimetics and glucose oxidase (GOx)/cholesterol oxidase (ChOx) in mesoporous silica to detect glucose and cholesterol in blood samples. The sensing mechanism involves the generation of H2O2 by the catalytic action of an immobilized oxidase on the target molecules in the sample. This subsequently activates the MNPs in the mesopores, thereby leading to the conversion of the substrate into a colored end product. This strategy is used to detect the target glucose or cholesterol molecules in the concentration range of 15-250 mg/dL. The response is highly linear and the lower detection limit is 7.5 mg/dL. The aforementioned colorimetric assay is extremely convenient, and it exhibits a high degree of linearity, precision, and reproducibility when employing real human blood samples. Therefore, this assay can be used in clinical practice for the multiplexed and reliable quantification of glucose and cholesterol. PMID:26726446

  15. Comparison of colorimetric methods for the quantification of model proteins in aqueous two-phase systems.

    PubMed

    Glyk, Anna; Heinisch, Sandra L; Scheper, Thomas; Beutel, Sascha

    2015-05-15

    In the current study, the quantification of different model proteins in the presence of typical aqueous two-phase system components was investigated by using the Bradford and bicinchoninic acid (BCA) assays. Each phase-forming component above 1 and 5 wt% had considerable effects on the protein quantification in both assays, respectively, resulting in diminished protein recoveries/absorption values by increasing poly(ethylene glycol) (PEG)/salt concentration and PEG molecular weight. Therefore, a convenient dilution of both components (up to 1 and 5 wt%) before protein quantification is recommended in both assays, respectively, where the BCA assay is favored in comparison with the Bradford assay. PMID:25684109

  16. Gold nanoparticle-based colorimetric aptasensor for rapid detection of six organophosphorous pesticides.

    PubMed

    Bai, Wenhui; Zhu, Chao; Liu, Jinchuan; Yan, Mengmeng; Yang, Shuming; Chen, Ailiang

    2015-10-01

    Fast immunoassay-based screening methods are unavailable for most small-molecule pesticides because of a lack of immunogenicity and the difficulty in obtaining antibodies by animal immunization. Aptamers are single-stranded DNA molecules selected through an in vitro process, which can bind to any target including nonimmunogenic small molecules with high affinity and specificity. Although various aptamer-based sensing methods have been developed for antibiotics, microorganisms, heavy metal ions, and biotoxins, there are few reports on aptamer-based methods for quick detection of organophosphorous pesticides. The gold (Au) nanoparticle (AuNP) colorimetric assay is a widely utilized rapid detection method because of properties such as easy operation and visualized results. In the present study, organophosphorous pesticide aptamers were adsorbed on the surface of AuNPs to stabilize the AuNP solution against high concentrations of salt to prevent AuNP aggregation. After the addition of targets, the aptamers binding to the targets are detached from the AuNPs, resulting in aggregation of AuNPs and a color change from red to purple-blue. The proposed method can detect 6 organophosphorous pesticides with good recoveries from 72% to 135% in environmental river water samples. The present study provides a new way for simple, rapid, and multiplex detection of organophosphorous pesticides. PMID:26031388

  17. Ultrasensitive electrospun fluorescent nanofibrous membrane for rapid visual colorimetric detection of H2O2.

    PubMed

    Senthamizhan, Anitha; Balusamy, Brabu; Aytac, Zeynep; Uyar, Tamer

    2016-02-01

    We report herein a flexible fluorescent nanofibrous membrane (FNFM) prepared by decorating the gold nanocluster (AuNC) on electrospun polysulfone nanofibrous membrane for rapid visual colorimetric detection of H2O2. The provision of AuNC coupled to NFM has proven to be advantageous for facile and quick visualization of the obtained results, permitting instant, selective, and on-site detection. We strongly suggest that the fast response time is ascribed to the enhanced probabilities of interaction with AuNC located at the surface of NF. It has been observed that the color change from red to blue is dependent on the concentration, which is exclusively selective for hydrogen peroxide. The detection limit has been found to be 500 nM using confocal laser scanning microscope (CLSM), visually recognizable with good accuracy and stability. A systematic comparison was performed between the sensing performance of FNFM and AuNC solution. The underlying sensing mechanism is demonstrated using UV spectra, transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS). The corresponding disappearance of the characteristic emissions of gold nanoclusters and the emergence of a localized surface plasmon resonance (LSPR) band, stressing this unique characteristic of gold nanoparticles. Hence, it is evident that the conversion of nanoparticles from nanoclusters has taken place in the presence of H2O2. Our work here has paved a new path for the detection of bioanalytes, highlighting the merits of rapid readout, sensitivity, and user-friendliness. PMID:26637215

  18. Field-deployable colorimetric biosensor system for the rapid detection of pathogenic organisms

    NASA Astrophysics Data System (ADS)

    Duy, Janice

    The rapid identification of pathogenic organisms is necessary for recognizing and managing human and environmental health risks. Numerous detection schemes are available, but most are difficult to employ in non-laboratory settings due to their need for bulky, specialized equipment, multiple reagents, or highly trained personnel. To address this problem, a rapid, field-compatible biosensor system based on the colorimetric detection of nucleic acid hybrids was developed. Peptide nucleic acid (PNA) probes were used to capture ribosomal RNA sequences from environmental samples. Non-target nucleic acids, including single-base mismatches flanked by adenines and uracils, were removed with a micrococcal nuclease digestion step. Matched PNA-RNA hybrids remained intact and were indicated by the cyanine dye DiSC2(5). PNA-containing duplexes function as templates for the aggregation of DiSC2(5), visualized as a change in solution color from blue to purple. This transition can be measured as an increase in the solution absorbance at 540 nm (dye aggregate) at the expense of the dye monomer peak at 650 nm. These concomitant spectral changes were used to calculate a "hybridization signal" using the ratio A aggregate/Amonomer ≈ A540/A650. Testing with pathogenic environmental samples was accomplished using two model organisms: the harmful algal bloom-causing dinoflagellate Alexandrium species, and the potato wart disease-causing fungus Synchytrium endobioticum. In both cases, the colorimetric approach was able to distinguish the targets with sensitivities rivaling those of established techniques, but with the advantages of decreased hands-on time and cost. Assay fieldability was tested with a portable colorimeter designed to quantify the dye-indicated hybridization signal and assembled from commercially available components. Side-by-side testing revealed no difference in the sensing performance of the colorimeter compared to a laboratory spectrophotometer (Pearson's r=0

  19. A colorimetric and ratiometric fluorescent probe for quantification of bovine serum albumin.

    PubMed

    Zeng, Xiaodan; Zhang, Xiaoling; Zhu, Baocun; Jia, Hongying; Li, Yamin; Xue, Juan

    2011-10-01

    A 4-aminonaphthalimide-based ratiometric fluorescent probe 1 employing the internal charge transfer (ICT) mechanism was designed and synthesized to detect bovine serum albumin (BSA). The interaction of 1 and BSA was investigated by fluorescence and UV-vis absorption spectroscopy. Upon treatment with BSA, the probe successfully exhibited a ratiometric fluorescent response at 540 nm and 480 nm. The fluorescent intensity ratio at 540 nm and 480 nm (F(540)/F(480)) increases linearly with BSA concentration in the range of 0-75.0 μg mL(-1) and the detection limit was about 2.4 ng mL(-1). Our strategy is expected to provide a methodology to quantify BSA either by a normal or by a ratiometric and colorimetric way with high sensitivity. PMID:21858298

  20. Screening the toxicity and biodegradability of petroleum hydrocarbons by a rapid colorimetric method.

    PubMed

    Montagnolli, Renato Nallin; Lopes, Paulo Renato Matos; Bidoia, Ederio Dino

    2015-02-01

    Crude oil and petroleum products have a wide variety of hazardous components with high toxicity and low biodegradability. Certain dyes change their colors by intercepting electron transfer reactions during the transformation processes. This study applied resazurin and 2,6-dichlorophenol-indophenol indicators for a rapid screening biodegradation capability and toxicity response to various petroleum products such as motor oil, diesel, gasoline, and phenol. Colorimetry tests were performed in test tubes, and the absorbance values were measured over time. We observed different discoloration profiles after degradation tests using Bacillus subtilis inoculum. Phytotoxicity assays were also performed to compare colorimetric screening assays with a conventional toxicity testing with plants (seed germination). The results indicated that biotransformation of oils can increase its overall toxicity. Intermediate byproducts can be formed through biodegradation and thereby increase the toxicity of oils. The assessment of acute toxicity has shown that phenol is extremely toxic to petroleum-biodegrading microbial communities. Low molecular-weight gasoline was considered biodegradable, but it also exhibited a high acute toxic effect, mainly due to its high solubility and the presence of more volatile compounds that can penetrate cells and potentially damage cellular structures. PMID:25537922

  1. Simple colorimetric method for quantification of surface carboxy groups on polymer particles.

    PubMed

    Hennig, Andreas; Hoffmann, Angelika; Borcherding, Heike; Thiele, Thomas; Schedler, Uwe; Resch-Genger, Ute

    2011-06-15

    We present a novel, simple, and fast colorimetric method to quantify the total number of carboxy groups on polymer microparticle and nanoparticle surfaces. This method exploits that small divalent transition metal cations (M(2+) = Ni(2+), Co(2+), Cd(2+)) are efficiently bound to these surface functional groups, which allows their extraction by a single centrifugation step. Remaining M(2+) in the supernatant is subsequently quantified spectrophotometrically after addition of the metal ion indicator pyrocatechol violet, for which Ni(2+) was identified to be the most suitable transition metal cation. We demonstrate that the difference between added and detected M(2+) is nicely correlated to the number of surface carboxy groups as determined by conductometry, thereby affording a validated measure for the trueness of this procedure. The variation coefficient of ~5% found in reproducibility studies underlines the potential of this novel method that can find conceivable applications for the characterization of different types of poly(carboxylic acid)-functionalized materials, e.g., for quality control by manufacturers of such materials. PMID:21561064

  2. Antimicrobial susceptibility of rapidly growing mycobacteria using the rapid colorimetric method.

    PubMed

    Ramis, I B; Cnockaert, M; von Groll, A; Nogueira, C L; Leão, S C; Andre, E; Simon, A; Palomino, J C; da Silva, P E A; Vandamme, P; Martin, A

    2015-07-01

    Drug susceptibility testing (DST) of rapidly growing mycobacteria (RGM) are recommended for guiding the antimicrobial therapy. We have evaluated the use of resazurin in Mueller-Hinton medium (MHR) for MIC determination of RGM and compared the results with those obtained with the reference standard broth microdilution in Mueller-Hinton (MH) and with the resazurin microtiter assay (REMA) in 7H9 broth. The MIC of eight drugs: amikacin (AMI), cefoxitin (FOX), ciprofloxacin (CIP), clarithromycin (CLA), doxycycline (DOX), linezolid (LZD), moxifloxacin (MXF) and trimethoprim-sulfamethoxazole (TMP-SMX) were evaluated against 76 RGM (18 species) using three methods (MH, MHR, and REMA) in a 96-well plate format incubated at 37 °C over 3-5 days. Results obtained in the MH plates were interpreted by the appearance of turbidity at the bottom of the well before adding the resazurin. MHR and 7H9-REMA plates were read by visual observation for a change in color from blue to pink. The majority of results were obtained at day 5 for MH and 1 day after for MHR and 7H9-REMA. However, the preliminary experiment on time to positivity results using the reference strain showed that the resazurin can be added to the MH at day 2 to produce the results at day 3, but future studies with large sets of strains are required to confirm this suggestion. A high level of agreement (kappa 1.000-0.884) was obtained between the MH and the MHR. Comparison of results obtained with 7H9-REMA, on the other hand, revealed several discrepancies and a lower level of agreement (kappa 1.000-0.111). The majority of the strains were resistant to DOX and TMP-SMX, and the most active antimicrobials for RGM were AMI and FOX. In the present study, MHR represented an excellent alternative for MIC determination of RGM. The results could be read reliably, more easily, and more quickly than with the classical MH method. PMID:25820290

  3. Rapid digital quantification of microfracture populations

    NASA Astrophysics Data System (ADS)

    Gomez, Leonel A.; Laubach, Stephen E.

    2006-03-01

    Populations of microfractures are a structural fabric in many rocks deformed at upper crustal conditions. In some cases these fractures are visible in transmitted-light microscopy as fluid-inclusion planes or cement filled microfractures, but because SEM-based cathodoluminescence (CL) reveals more fractures and delineates their shapes, sizes, and crosscutting relations, it is a more effective structural tool. Yet at magnifications of 150-300×, at which many microfractures are visible, SEM-CL detectors image only small sample areas (0.5-0.1 mm 2) relative to fracture population patterns. The substantial effort required to image and measure centimeter-size areas at high-magnification has impeded quantitative study of microfractures. We present a method for efficient collection of mosaics of high-resolution CL imagery, a preparation method that allows samples to be any size while retaining continuous imagery of rock (no gaps), and software that facilitates fracture mapping and data reduction. Although the method introduced here was developed for CL imagery, it can be used with any other kind of images, including mosaics from petrographic microscopes. Compared with manual measurements, the new method increases several fold the number of microfractures imaged without a proportional increase in level of effort, increases the accuracy and repeatability of fracture measurements, and speeds quantification and display of fracture population attributes. We illustrate the method on microfracture arrays in dolostone from NE Mexico and sandstone from NW Scotland. We show that key aspects of microfracture population attributes are only fully manifest at scales larger than a single thin section.

  4. Straightforward rapid spectrophotometric quantification of total cyanogenic glycosides in fresh and processed cassava products.

    PubMed

    Tivana, Lucas Daniel; Da Cruz Francisco, Jose; Zelder, Felix; Bergenståhl, Bjorn; Dejmek, Petr

    2014-09-01

    In this study, we extend pioneering studies and demonstrate straightforward applicability of the corrin-based chemosensor, aquacyanocobyrinic acid (ACCA), for the instantaneous detection and rapid quantification of endogenous cyanide in fresh and processed cassava roots. Hydrolytically liberated endogenous cyanide from cyanogenic glycosides (CNp) reacts with ACCA to form dicyanocobyrinic acid (DCCA), accompanied by a change of colour from orange to violet. The method was successfully tested on various cassava samples containing between 6 and 200 mg equiv. HCN/kg as verified with isonicotinate/1,3-dimethylbarbiturate as an independent method. The affinity of ACCA sensor to cyanide is high, coordination occurs fast and the colorimetric response can therefore be instantaneously monitored with spectrophotometric methods. Direct applications of the sensor without need of extensive and laborious extraction processes are demonstrated in water-extracted samples, in acid-extracted samples, and directly on juice drops. ACCA showed high precision with a standard deviation (STDV) between 0.03 and 0.06 and high accuracy (93-96%). Overall, the ACCA procedure is straightforward, safe and easily performed. In a proof-of-concept study, rapid screening of ten samples within 20 min has been tested. PMID:24731309

  5. A smartphone-based colorimetric reader coupled with a remote server for rapid on-site catechols analysis.

    PubMed

    Wang, Yun; Li, Yuanyuan; Bao, Xu; Han, Juan; Xia, Jinchen; Tian, Xiaoyu; Ni, Liang

    2016-11-01

    The search of a practical method to analyze cis-diol-containing compounds outside laboratory settings remains a substantial scientific challenge. Herein, a smartphone-based colorimetric reader was coupled with a remote server for rapid on-site analysis of catechols. A smallest-scale 2×2 colorimetric sensor array composed of pH indicators and phenylboronic acid was configured. The array was able to distinguish 13 catechols at 6 serial concentrations, through simultaneous treatment via principal component analysis, hierarchical cluster analysis, and linear discriminant analysis. After both the discriminatory power of the array and the prediction ability of the partial least squares quantitative models were proved to be predominant, the smartphone was coupled to the remote server. All the ΔRGB data were uploaded to the remote server wherein linear discriminant analysis and partial least squares processing modules were established to provide qualitative discrimination and quantitative calculation, respectively, of the analytes in real time. The applicability of this novel method to a real-life scenario was confirmed by the on-site analysis of various catechols from a water sample of the Yangtze River; the feedback result in the smartphone showed the method was able to identify the catechols with 100% accuracy and predict the concentrations to within 0.706-2.240 standard deviation. PMID:27591604

  6. The Rapid and Sensitive Quantitative Determination of Galactose by Combined Enzymatic and Colorimetric Method: Application in Neonatal Screening.

    PubMed

    Kianmehr, Anvarsadat; Mahrooz, Abdolkarim; Ansari, Javad; Oladnabi, Morteza; Shahbazmohammadi, Hamid

    2016-05-01

    The quantitative measurement of galactose in blood is essential for the early diagnosis, treatment, and dietary monitoring of galactosemia patients. In this communication, we aimed to develop a rapid, sensitive, and cost-effective combined method for galactose determination in dry blood spots. This procedure was based on the combination of enzymatic reactions of galactose dehydrogenase (GalDH), dihydrolipoyl dehydrogenase (DLD), and alkaline phosphates with a colorimetric system. The incubation time and the concentration of enzymes used in new method were also optimized. The analytical performance was studied by the precision, recovery, linearity, and sensitivity parameters. Statistical analysis was applied to method comparison experiment. The regression equation and correlation coefficient (R (2)) were Y = 0.0085x + 0.032 and R (2) = 0.998, respectively. This assay exhibited a recovery in the range of 91.7-114.3 % and had the limit detection of 0.5 mg/dl for galactose. The between-run coefficient of variation (CV) was between 2.6 and 11.1 %. The within-run CV was between 4.9 and 9.2 %. Our results indicated that the new and reference methods were in agreement because no significant biases exist between them. Briefly, a quick and reliable combined enzymatic and colorimetric assay was presented for application in newborn mass screening and monitoring of galactosemia patients. PMID:26821257

  7. Centrifugal loop-mediated isothermal amplification microdevice for rapid, multiplex and colorimetric foodborne pathogen detection.

    PubMed

    Oh, Seung Jun; Park, Byung Hyun; Jung, Jae Hwan; Choi, Goro; Lee, Doh C; Kim, Do Hyun; Seo, Tae Seok

    2016-01-15

    We present a centrifugal microfluidic device which enables multiplex foodborne pathogen identification by loop-mediated isothermal amplification (LAMP) and colorimetric detection using Eriochrome Black T (EBT). Five identical structures were designed in the centrifugal microfluidic system to perform the genetic analysis of 25 pathogen samples in a high-throughput manner. The sequential loading and aliquoting of the LAMP cocktail, the primer mixtures, and the DNA sample solutions were accomplished by the optimized zigzag-shaped microchannels and RPM control. We targeted three kinds of pathogenic bacteria (Escherichia coli O157:H7, Salmonella typhimurium and Vibrio parahaemolyticus) and detected the amplicons of the LAMP reaction by the EBT-mediated colorimetric method. For the limit-of-detection (LOD) test, we carried out the LAMP reaction on a chip with serially diluted DNA templates of E. coli O157:H7, and could observe the color change with 380 copies. The used primer sets in the LAMP reaction were specific only to the genomic DNA of E. coli O157:H7, enabling the on-chip selective, sensitive, and high-throughput pathogen identification with the naked eyes. The entire process was completed in 60min. Since the proposed microsystem does not require any bulky and expensive instrumentation for end-point detection, our microdevice would be adequate for point-of-care (POC) testing with high simplicity and high speed, providing an advanced genetic analysis microsystem for foodborne pathogen detection. PMID:26322592

  8. Rapid fluorometric quantification of bacterial cells using Redsafe nucleic acid stain

    PubMed Central

    Khalili, Ehsan; Hosseini, Vahid; Solhi, Roya; Aminian, Mahdi

    2015-01-01

    Background and Objectives: Numerous procedures in biology and medicine require the counting of cells. Direct enumeration of Colony Forming Units (CFUs) is time-consuming and dreary accurate cell counting on plates with high numbers of CFUs is error prone. In this study we report a new indirect cell counting method that was developed based on the use of Redsafe fluorometric assay. The usefulness of Redsafe, a nucleic acid stain, in liquid medium is based on the binding of the fluorescent dye to DNA. Materials and Methods: Redsafe fluorometric assay was evaluated in comparison with MTT colorimetric assay as a colourimetric assay for enumeration of bacterial cells. Results: Obtained results showed that fluorometric assay threshold for LB grown E. coli is 6×104 CFU/ml. Redsafe fluorescent assay can be used as a rapid and inexpensive method for bacterial enumeration and quantification with increased sensitivity. Conclusion: The sensitivity of the Redsafe fluorometric assay for detection and enumeration of bacterial cells was 2-log-unit more than that was observed for the MTT assay. PMID:26885332

  9. O-antigen and virulence profiling of Shiga toxin-producing Escherichia coli by a rapid and cost-effective DNA microarray colorimetric method

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Shiga toxin-producing Escherichia coli (STEC) is a leading cause of foodborne illness worldwide. To evaluate better methods to rapidly detect and genotype Shiga toxin-producing Escherichia coli strains, the present study evaluated the use of the ampliPHOX colorimetric detection technology, based on ...

  10. Rapid quantification method for Legionella pneumophila in surface water.

    PubMed

    Wunderlich, Anika; Torggler, Carmen; Elsässer, Dennis; Lück, Christian; Niessner, Reinhard; Seidel, Michael

    2016-03-01

    World-wide legionellosis outbreaks caused by evaporative cooling systems have shown that there is a need for rapid screening methods for Legionella pneumophila in water. Antibody-based methods for the quantification of L. pneumophila are rapid, non-laborious, and relatively cheap but not sensitive enough for establishment as a screening method for surface and drinking water. Therefore, preconcentration methods have to be applied in advance to reach the needed sensitivity. In a basic test, monolithic adsorption filtration (MAF) was used as primary preconcentration method that adsorbs L. pneumophila with high efficiency. Ten-liter water samples were concentrated in 10 min and further reduced to 1 mL by centrifugal ultrafiltration (CeUF). The quantification of L. pneumophila strains belonging to the monoclonal subtype Bellingham was performed via flow-based chemiluminescence sandwich microarray immunoassays (CL-SMIA) in 36 min. The whole analysis process takes 90 min. A polyclonal antibody (pAb) against L. pneumophila serogroup 1-12 and a monoclonal antibody (mAb) against L. pneumophila SG 1 strain Bellingham were immobilized on a microarray chip. Without preconcentration, the detection limit was 4.0 × 10(3) and 2.8 × 10(3) CFU/mL determined by pAb and mAb 10/6, respectively. For samples processed by MAF-CeUF prior to SMIA detection, the limit of detection (LOD) could be decreased to 8.7 CFU/mL and 0.39 CFU/mL, respectively. A recovery of 99.8 ± 15.9% was achieved for concentrations between 1-1000 CFU/mL. The established combined analytical method is sensitive for rapid screening of surface and drinking water to allow fast hygiene control of L. pneumophila. PMID:26873217

  11. Colorimetric method for rapid detection of Oxacillin resistance in Staphylococcus aureus and its comparison with PCR for mec A gene

    PubMed Central

    Ghanwate, Niraj; Thakare, Prashant; Bhise, P. R.; Gawande, Sonali

    2016-01-01

    Rapid and accurate detection of Methicillin Resistant Staphylococcus aureus (MRSA) is an important role of clinical microbiology laboratories to avoid treatment failure. The detection of MRSA is based on phenotypic assays which require at least 24 h to perform. Detection of the mecA gene or of PBP 2a is the “gold standard”, but not always available. The aim of this study was to evaluate a rapid method for detection of MRSA by using 3 (4, 5 dimethyl thiazole -2-yl) -2, 5 diphenyl tetrazolium bromide (MTT). Total 126 isolates of MRSA were collected from tertiary healthcare center and were confirmed by oxacillin screening agar test as per CLSI guidelines. Amplification of mecA gene was performed by using PCR. MTT assay was carried out for all the isolates in 96 well Microtitre plate and compared with standard methods of CLSI. Out of 126 isolates, 98 were found to be mecA positive. MTT method was found to be 98.98% sensitive and 96.43% specific. The MTT based colorimetric method is rapid and simple test for screening of oxacillin resistance in Staphylococcus aureus. It significantly shortens the time to just 7 h required to obtained a drug susceptibility test and could be useful to screen MRSA. PMID:26960268

  12. Colorimetric method for rapid detection of Oxacillin resistance in Staphylococcus aureus and its comparison with PCR for mec A gene.

    PubMed

    Ghanwate, Niraj; Thakare, Prashant; Bhise, P R; Gawande, Sonali

    2016-01-01

    Rapid and accurate detection of Methicillin Resistant Staphylococcus aureus (MRSA) is an important role of clinical microbiology laboratories to avoid treatment failure. The detection of MRSA is based on phenotypic assays which require at least 24 h to perform. Detection of the mecA gene or of PBP 2a is the "gold standard", but not always available. The aim of this study was to evaluate a rapid method for detection of MRSA by using 3 (4, 5 dimethyl thiazole -2-yl) -2, 5 diphenyl tetrazolium bromide (MTT). Total 126 isolates of MRSA were collected from tertiary healthcare center and were confirmed by oxacillin screening agar test as per CLSI guidelines. Amplification of mecA gene was performed by using PCR. MTT assay was carried out for all the isolates in 96 well Microtitre plate and compared with standard methods of CLSI. Out of 126 isolates, 98 were found to be mecA positive. MTT method was found to be 98.98% sensitive and 96.43% specific. The MTT based colorimetric method is rapid and simple test for screening of oxacillin resistance in Staphylococcus aureus. It significantly shortens the time to just 7 h required to obtained a drug susceptibility test and could be useful to screen MRSA. PMID:26960268

  13. Rapid enumeration of Fecal Coliforms in water by a colorimetric beta-galactosidase assay.

    PubMed Central

    Warren, L S; Benoit, R E; Jessee, J A

    1978-01-01

    The colorimetric beta-galactosidase assay is based upon the enzymatic hydrolysis of the substrate o-nitrophenyl-beta-D-galactoside (ONPG) by fecal coliforms. This technique provides an estimate of the fecal coliform concentration within 8 to 20 h. A 100-ml portion of test sample was passed through a 0.45 micrometer membrane filter. This filter was then incubated at 37 degrees C for 1 h in EC medium followed by the addition of filter-sterilized ONPG. The incubation was continued at 44.5 degrees C until a half-maximum absorbance (at 420 nm) was reached. The time between the start of incubation and the half-maximum absorbance was proportional to the concentration of fecal coliforms present. Escherichia coli (K-12) was used to measure the kinetics of substrate hydrolysis and the response time of different cell concentrations. High cell densities produced an immediate response, whereas 1 cell/ml will produce a response in less than 20 h. In field studies in which samples were taken from a range of grossly polluted streams to relatively clean lake water, a linear correlation between ONPG hydrolysis times and fecal coliform most-probable-number values was established. A total of 302 isolates randomly selected from positive ONPG-EC media, which were derived from 11 different habitats, were identified as E. coli (96.69 percent), Enterobacter cloacae (2.32 percent), Klebsiella pneumoniae (0.66 percent), and Citrobacter freundii (0.33 percent). PMID:414659

  14. Rapid quantitative analysis of lipids using a colorimetric method in a microplate format.

    PubMed

    Cheng, Yu-Shen; Zheng, Yi; VanderGheynst, Jean S

    2011-01-01

    A colorimetric sulfo-phospho-vanillin (SPV) method was developed for high throughput analysis of total lipids. The developed method uses a reaction mixture that is maintained in a 96-well microplate throughout the entire assay. The new assay provides the following advantages over other methods of lipid measurement: (1) background absorbance can be easily corrected for each well, (2) there is less risk of handling and transferring sulfuric acid contained in reaction mixtures, (3) color develops more consistently providing more accurate measurement of absorbance, and (4) the assay can be used for quantitative measurement of lipids extracted from a wide variety of sources. Unlike other spectrophotometric approaches that use fluorescent dyes, the optimal spectra and reaction conditions for the developed assay do not vary with the sample source. The developed method was used to measure lipids in extracts from four strains of microalgae. No significant difference was found in lipid determination when lipid content was measured using the new method and compared to results obtained using a macro-gravimetric method. PMID:21069472

  15. Genetically engineered colorimetric single-chain antibody fusion protein for rapid diagnosis of rabies virus.

    PubMed

    Mousli, M; Turki, I; Kharmachi, H; Dellagi, K

    2008-01-01

    The most widely used test for rabies diagnostics is the fluorescent antibody test, which is recommended by both the World Health Organization and the World Organisation for Animal Health (OIE). This test may be used directly on a smear, and can also be used to confirm the presence of rabies antigen in cell culture or in brain tissue for diagnosis. The colorimetric enzymes are usually coupled to an antibody by chemical means using cross-linking reagents. However, such non-specific procedures lead to heterogeneous conjugates, sometimes with reduced activity and specificity. To bypass these problems, genetic engineering has provided a way to create chimeric bifunctional molecules in which the variable domains of an antibody are genetically linked to unrelated protein tracers. In this study, we describe the successful production of a bifunctional chimeric protein based on alkaline phosphatase-fused anti-rabies virus glycoprotein scFv antibody fragment. We also report the antigen binding properties and the alkaline phosphatase activity of the recombinant conjugate protein. We established its value as a novel in vitro tool for detecting the rabies virus in brain smear in a one-step procedure; it presents a similar sensitivity and specificity to that obtained using standard reagents. PMID:18634511

  16. Rapid susceptibility testing for slowly growing nontuberculous mycobacteria using a colorimetric microbial viability assay based on the reduction of water-soluble tetrazolium WST-1.

    PubMed

    Tsukatani, T; Suenaga, H; Shiga, M; Ikegami, T; Ishiyama, M; Ezoe, T; Matsumoto, K

    2015-10-01

    Rapid susceptibility testing for slowly growing nontuberculous mycobacteria (NTM) using a colorimetric microbial viability assay based on the reduction of the water-soluble tetrazolium salt {2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-1)} using 2,3,5,6-tetramethyl-1,4-benzoquinone as an electron mediator was developed. Using the Clinical and Laboratory Standards Institute (CLSI) method, a long-term incubation time (7-14 days) was required to determine the minimum inhibitory concentrations (MICs) of the slowly growing NTM. The MICs for a variety of different antibiotics against the slowly growing NTM were determined by the WST-1 colorimetric method and compared with those obtained using the broth microdilution methods approved by the CLSI. Good agreement was found between the MICs determined after 3-4 days using the WST-1 colorimetric method and those obtained after 10-14 days using the broth microdilution method. The results suggest that the WST-1 colorimetric assay is a useful method for the rapid determination of the MICs for the slowly growing NTM. PMID:26173690

  17. Integration of Novel Low-Cost Colorimetric, Laser Photometric, and Visual Fluorescent Techniques for Rapid Identification of Falsified Medicines in Resource-Poor Areas: Application to Artemether–Lumefantrine

    PubMed Central

    Green, Michael D.; Hostetler, Dana M.; Nettey, Henry; Swamidoss, Isabel; Ranieri, Nicola; Newton, Paul N.

    2015-01-01

    The availability of falsified antimalarial drugs can be reduced with effective drug regulatory agencies and proper enforcement. Fundamental to these agencies taking action, rapid identification must be made as soon as they appear in the market place. Since falsified antimalarials occur mostly in developing countries, performing drug analysis presents itself with unique challenges. A fundamental factor in choosing a useful technique is affordability and simplicity. Therefore, we suggest a three-tiered drug evaluation strategy for identifying a falsified drug in resource-poor areas. Tier I is a simple comparison of a tablet's weight and dimensions with official specifications. Tier II uses inexpensive photometric devices (laser and fluorescence) to evaluate a tablet. Suspicious samples from Tier I and II assessments are then subjected to a colorimetric assay for active ingredients identification and quantification. In this article, we evaluate a novel colorimetric assay for the simultaneous assessment of both lumefantrine and artemether in co-formulated Coartem™ tablets, and integrate the method with two novel, low-cost, fluorescence and laser photometric devices. Image analysis software is used for the assessments. Although artemether–lumefantrine is used as an example, the strategy may be adapted to other medicines. PMID:25897066

  18. Rapid method for the quantification of hydroquinone concentration: chemiluminescent analysis.

    PubMed

    Chen, Tung-Sheng; Liou, Show-Yih; Kuo, Wei-Wen; Wu, Hsi-Chin; Jong, Gwo-Ping; Wang, Hsueh-Fang; Shen, Chia-Yao; Padma, V Vijaya; Huang, Chih-Yang; Chang, Yen-Lin

    2015-11-01

    Topical hydroquinone serves as a skin whitener and is usually available in cosmetics or on prescription based on the hydroquinone concentration. Quantification of hydroquinone content therefore becomes an important issue in topical agents. High-performance liquid chromatography (HPLC) is the commonest method for determining hydroquinone content in topical agents, but this method is time-consuming and uses many solvents that can become an environmental issue. We report a rapid method for quantifying hydroquinone content by chemiluminescent analysis. Hydroquinone induces the production of hydrogen peroxide in the presence of basic compounds. Hydrogen peroxide induced by hydroquinone oxidized light-emitting materials such as lucigenin, resulted in the production of ultra-weak chemiluminescence that was detected by a chemiluminescence analyzer. The intensity of the chemiluminescence was found to be proportional to the hydroquinone concentration. We suggest that the rapid (measurement time, 60 s) and virtually solvent-free (solvent volume, <2 mL) chemiluminescent method described here for quantifying hydroquinone content may be an alternative to HPLC analysis. PMID:25693839

  19. Colorimetric quantification of mRNA expression in rare tumour cells amplified by multiple ligation-dependent probe amplification.

    PubMed

    Acero Sanchez, Josep L; Henry, Olivier Y F; Mairal, Teresa; Laddach, Nadja; Nygren, Anders; Hauch, Siegfried; Fetisch, Jasmin; O'Sullivan, Ciara K

    2010-07-01

    An enzyme-linked oligonucleotide assay (ELONA) for quantification of mRNA expression of five genes involved in breast cancer, extracted from isolated rare tumour cells and amplified by multiplex ligation-dependent probe amplification (MLPA) is presented. In MLPA, a multiplex oligonucleotide ligation assay is combined with a PCR reaction in which all ligation products are amplified by use of a single primer pair. Biotinylated probes complementary to each of the target sequences were immobilised on the surface of a streptavidin-coated microtitre plate and exposed to single-stranded MLPA products. A universal reporting probe sequence modified with horseradish peroxidase (URP-HRP) and complementary to a universal primer used during the MLPA step was further added to the surface-bound duplex as a reporter probe. Simultaneous addition of anchoring probe and target, followed by addition of reporter probe, rather than sequential addition, was achieved with no significant effect on sensitivity and limits of detection, but considerably reduced the required assay time. Detection limits as low as 20 pmol L(-1), with an overall assay time of 95 min could be achieved with negligible cross-reactivity between probes and non-specific targets present in the MLPA-PCR product. The same MLPA-PCR product was analysed using capillary electrophoresis, the technique typically used for analysis of MLPA products, and good correlation was observed. The assay presented is easy to carry out, relatively inexpensive, rapid, does not require sophisticated instrumentation, and enables quantitative analysis, making it very promising for the analysis of MLPA products. PMID:20526769

  20. Fe3O4 magnetic nanoparticle peroxidase mimetic-based colorimetric assay for the rapid detection of organophosphorus pesticide and nerve agent.

    PubMed

    Liang, Minmin; Fan, Kelong; Pan, Yong; Jiang, Hui; Wang, Fei; Yang, Dongling; Lu, Di; Feng, Jing; Zhao, Jianjun; Yang, Liu; Yan, Xiyun

    2013-01-01

    Rapid and sensitive detection methods are in urgent demand for the screening of extensively used organophosphorus pesticides and highly toxic nerve agents for their neurotoxicity. In this study, we developed a novel Fe(3)O(4) magnetic nanoparticle (MNP) peroxidase mimetic-based colorimetric method for the rapid detection of organophosphorus pesticides and nerve agents. The detection assay is composed of MNPs, acetylcholinesterase (AChE), and choline oxidase (CHO). The enzymes AChE and CHO catalyze the formation of H(2)O(2) in the presence of acetylcholine, which then activates MNPs to catalyze the oxidation of colorimetric substrates to produce a color reaction. After incubation with the organophosphorus neurotoxins, the enzymatic activity of AChE was inhibited and produced less H(2)O(2), resulting in a decreased catalytic oxidation of colorimetric substrates over MNP peroxidase mimetics, accompanied by a drop in color intensity. Three organophosphorus compounds were tested on the assay: acephate and methyl-paraoxon as representative organophosphorus pesticides and the nerve agent Sarin. The novel assay displayed substantial color change after incubation in organophosphorus neurotoxins in a concentration-dependent manner. As low as 1 nM Sarin, 10 nM methyl-paraoxon, and 5 μM acephate are easily detected by the novel assay. In conclusion, by employing the peroxidase-mimicking activity of MNPs, the developed colorimetric assay has the potential of becoming a screening tool for the rapid and sensitive assessment of the neurotoxicity of an overwhelming number of organophosphate compounds. PMID:23153113

  1. Colorimetric quantification of sucrose in presence of thermo-sensitive polymers present in aqueous two-phase systems.

    PubMed

    Ramalakshmi, Subbarayalu; Ooi, Chien Wei; Ariff, Arbakariya B; Ramanan, Ramakrishnan Nagasundara

    2014-01-01

    The use of biodegradable material such as simple carbohydrates and recyclable material such as thermo-sensitive polymers is in need to develop a sustainable aqueous two-phase system (ATPS) for the purification of biomolecules. Accurate determination of sucrose concentration is important in liquid-liquid equilibrium (LLE) study of carbohydrate-based ATPS. The well-established phenol-sulfuric acid method has been widely employed in the measurement of carbohydrate concentration. However, the presence of thermo-sensitive polymers, which has a lower critical solution temperature (LCST) below room temperature, in carbohydrate samples could hamper the precision of spectrophotometric analysis due to the formation of two phases or cloudiness in the sample. Thus, the following modifications were made in an attempt to eliminate the interference occurred during conventional phenol-sulfuric acid assay.•The modified assay for sucrose quantification was performed at an ice-cold temperature throughout the reaction in order to avoid the interference from thermo-sensitive polymers.•This method required a sample volume of 3 μL and hence the volume of other reagents employed was also considerably reduced.•The absorbance was measured at 520 nm which allowed a longer linearity range (0.05-7.5%, w/v). PMID:26150957

  2. Rapid detection and quantification of impact damage in composite structures

    NASA Technical Reports Server (NTRS)

    Smith, Barry T.

    1992-01-01

    It is shown that a multidisciplinary nondestructive evaluation approach for impact damage detection in composite structures can be used to produce a more efficient inspection. The multidisciplinary NDE approach relies on fast large area thermographic inspections along with detailed ultrasonic volumetric imaging. The thermal inspection technique rapidly identifies the impact damage. The ultrasonic volumetric imaging quantifies the impact generated delaminations through the volume of the structure.

  3. Rapid colorimetric assay of urinary beta-galactosidase and N-acetyl-beta-D-glucosaminidase with Cobas Mire Auto-analyzer.

    PubMed

    Xu, G; Zhu, L; Hong, J; Cao, Y; Xia, T

    1999-01-01

    A rapid and simple colorimetric method for the assay of urinary beta-galactosidase (GAL) and N-acetyl-beta-D-glucosaminidase (NAG) using 4-nitrophenyl-glycosides as the substrates and a Cobas Mire Auto-analyzer is described. Optimal conditions for this method including substrate concentration, ionic strength, pH and incubation time were characterized. Endogenous substances in urine did not interfere with the assays. Incubation time could be shortened to 10 minutes. A small volume of an alkaline buffer to terminate the enzyme reaction achieved better sensitivity and accuracy. Buffer-substrate solutions were stable for at least one week at 4 degrees C. Normal values of the two urinary enzymes are also reported. PMID:10323472

  4. Rapid quantification of soilborne pathogen communities in wheat-based long-term field experiments

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Traditional isolation and quantification of inoculum density is difficult for most soilborne pathogens. Quantitative PCR methods have been developed to rapidly identify and quantify many of these pathogens using a single DNA extract from soil. Rainfed experiments operated continuously for up to 84 y...

  5. Simple and rapid quantification of thrombocytes in zebrafish larvae.

    PubMed

    Huarng, Michael C; Shavit, Jordan A

    2015-06-01

    Platelets are a critical component of hemostasis, with disorders of number or function resulting in coagulation disturbances. Insights into these processes have primarily been realized through studies using mammalian models or tissues. Increasingly, zebrafish embryos and larvae have been used to study the protein and cellular components of hemostasis and thrombosis, including the thrombocyte, a nucleated platelet analog. However, investigations of thrombocytes have been somewhat limited due to lack of a robust and simple methodology for quantitation, an important component of platelet studies in mammals. Using video capture, we have devised an assay that produces a rapid, reproducible, and precise measurement of thrombocyte number in zebrafish larvae by counting fluorescently tagged cells. Averaging 1000 frames, we were able to subtract background fluorescence, thus limiting assessment to circulating thrombocytes. This method facilitated rapid assessment of relative thrombocyte counts in a population of 372 zebrafish larvae by a single operator in less than 3 days. This technique requires basic microscopy equipment and rudimentary programming, lends itself to high throughput analysis, and will enhance future studies of thrombopoiesis in the zebrafish. PMID:25790244

  6. Graphene-based rapid and highly-sensitive immunoassay for C-reactive protein using a smartphone-based colorimetric reader.

    PubMed

    Vashist, Sandeep Kumar; Marion Schneider, E; Zengerle, Roland; von Stetten, Felix; Luong, John H T

    2015-04-15

    A novel immunoassay (IA) has been developed for human C-reactive protein (CRP), an important biomarker and tissue preserving factor for infection and inflammation. Graphene nanoplatelets (GNP) and 3-aminopropyltriethoxysilane (APTES) were admixed and covalently attached to a polystyrene based-microtiter plate (MTP), pretreated with KOH. The resulting surface served as a stable layer for the covalent attachment of the anti-human CRP antibody. The IA procedure was based on the one-step kinetics-based sandwich IA employing a minimum number of process steps, whereas the enzymatic reaction solution was monitored by a smartphone-based colorimetric reader. With a limit of detection and a limit of quantification of 0.07ngmL(-1) and 0.9ngmL(-1), it precisely detected CRP spiked in diluted human whole blood and plasma as well as the CRP levels in clinical plasma samples. The results obtained for "real-world" patient samples agreed well with those of the conventional immunosorbent assay and the clinically-accredited analyzer-based IA. The antibody-bound GNP-functionalized MTPs retained its original activity after 6 weeks of storage in 0.1M PBS, pH 7.4 at 4°C. PMID:25460898

  7. A rapid molecular diagnosis of cutaneous leishmaniasis by colorimetric malachite green-loop-mediated isothermal amplification (LAMP) combined with an FTA card as a direct sampling tool.

    PubMed

    Nzelu, Chukwunonso O; Cáceres, Abraham G; Guerrero-Quincho, Silvia; Tineo-Villafuerte, Edwin; Rodriquez-Delfin, Luis; Mimori, Tatsuyuki; Uezato, Hiroshi; Katakura, Ken; Gomez, Eduardo A; Guevara, Angel G; Hashiguchi, Yoshihisa; Kato, Hirotomo

    2016-01-01

    Leishmaniasis remains one of the world's most neglected diseases, and early detection of the infectious agent, especially in developing countries, will require a simple and rapid test. In this study, we established a quick, one-step, single-tube, highly sensitive loop-mediated isothermal amplification (LAMP) assay for rapid detection of Leishmania DNA from tissue materials spotted on an FTA card. An FTA-LAMP with pre-added malachite green was performed at 64°C for 60min using a heating block and/or water bath and DNA amplification was detected immediately after incubation. The LAMP assay had high detection sensitivity down to a level of 0.01 parasites per μl. The field- and clinic-applicability of the colorimetric FTA-LAMP assay was demonstrated with 122 clinical samples collected from patients suspected of having cutaneous leishmaniasis in Peru, from which 71 positives were detected. The LAMP assay in combination with an FTA card described here is rapid and sensitive, as well as simple to perform, and has great potential usefulness for diagnosis and surveillance of leishmaniasis in endemic areas. PMID:26516109

  8. Gold nanoprobe functionalized with specific fusion protein selection from phage display and its application in rapid, selective and sensitive colorimetric biosensing of Staphylococcus aureus.

    PubMed

    Liu, Pei; Han, Lei; Wang, Fei; Petrenko, Valery A; Liu, Aihua

    2016-08-15

    Staphylococcus aureus (S. aureus) is one of the most ubiquitous pathogens in public healthcare worldwide. It holds great insterest in establishing robust analytical method for S. aureus. Herein, we report a S. aureus-specific recognition element, isolated from phage monoclone GQTTLTTS, which was selected from f8/8 landscape phage library against S. aureus in a high-throughput way. By functionalizing cysteamine (CS)-stabilized gold nanoparticles (CS-AuNPs) with S. aureus-specific pVIII fusion protein (fusion-pVIII), a bifunctional nanoprobe (CS-AuNPs@fusion-pVIII) for S. aureus was developed. In this strategy, the CS-AuNPs@fusion-pVIII could be induced to aggregate quickly in the presence of target S. aureus, resulting in a rapid colorimetric response of gold nanoparticles. More importantly, the as-designed probe exhibited excellent selectivity over other bacteria. Thus, the CS-AuNPs@fusion-pVIII could be used as the indicator of target S. aureus. This assay can detect as low as 19CFUmL(-1)S. aureus within 30min. Further, this approach can be applicable to detect S. aureus in real water samples. Due to its sensitivity, specificity and rapidness, this proposed method is promising for on-site testing of S. aureus without using any costly instruments. PMID:27085951

  9. A facile strategy for the preparation of ZnS nanoparticles deposited on montmorillonite and their higher catalytic activity for rapidly colorimetric detection of H2O2.

    PubMed

    Ding, Yanyuan; Sun, Lifang; Jiang, Yanling; Liu, Shunxiang; Chen, Mingxing; Chen, Miaomiao; Ding, Yanan; Liu, Qingyun

    2016-10-01

    In this paper, ZnS nanoparticles deposited on montmorillonite (ZnS-MMT) were prepared by a facile method at room temperature and characterized by powder X-ray diffraction (XRD), Energy-dispersive X-ray Detector (EDX) and transmission electron microscope (TEM), respectively. Significantly, the as-prepared ZnS-MMT nanocomposites have been proven to possess intrinsic peroxidase-like activity that can rapidly catalyze the reaction of peroxidase substrate 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2 and produce a blue color product in less than 30 seconds, which provides a sensitive colorimetric sensor to detect H2O2. Due to the synergistic effects between montmorillonite and ZnS nanoparticles, the obtained ZnS-MMT nanocomposites exhibit higher catalytic activity than that of MMT or ZnS alone. The catalytic behaviors of the ZnS-MMT nanocomposites showed a typical Michaelis-Menten kinetics. The catalytic activity and the catalytic mechanism were investigated using the procedures of steady-state kinetics and hydroxyl radical detection. ESR data revealed that the peroxidase-like activity of ZnS-MMT originated from the generation of OH radicals. PMID:27287113

  10. Rapid Identification and Drug Susceptibility Testing of Mycobacterium tuberculosis: Standard Operating Procedure for Non-Commercial Assays: Part 3: Colorimetric Redox Indicator Assay v1.3.12

    PubMed Central

    Singh, Sarman; Kumar, Parveen; Sharma, Shreya; Mumbowa, Francis; Martin, Anandi; Durier, Nicolas

    2012-01-01

    The previous two standard operating procedures (SOPs) related to the culture and drug susceptibility testing (DST) of Mycobacterium tuberculosis with the microscopic observation drug susceptibility assay (Part 1) and nitrate reductase assay (Part 2). The present SOP is devoted to a third non-commercial culture and DST method known as colorimetric redox indicator assay (CRI). As its name indicates, the CRI detects the ability of the M. tuberculosis to reduce the colored oxidation-reduction indicator when added to a liquid culture of M. tuberculosis, after exposing the growth to different anti-mycobacterial drugs. The change in the color of the indicator denotes the proportionate number of viable Mycobacteria in the medium. The identification and DST results can be obtained in 7-8 days. This SOP document has been developed through the culture and DST subgroup of the STOP tuberculosis (TB) Partnership New Diagnostic Working Group. It is intended for laboratories that would want to use or already use this rapid non-commercial method for culture identification and DST of M. tuberculosis, notably in resource-constraint settings in Asia and Africa. PMID:23440615

  11. Integration of Nanoparticle-Based Paper Sensors into the Classroom: An Example of Application for Rapid Colorimetric Analysis of Antioxidants

    ERIC Educational Resources Information Center

    Sharpe, Erica; Andreescu, Silvana

    2015-01-01

    We describe a laboratory experiment that employs the Nanoceria Reducing Antioxidant Capacity (or NanoCerac) Assay to introduce students to portable nanoparticle-based paper sensors for rapid analysis and field detection of polyphenol antioxidants. The experiment gives students a hands-on opportunity to utilize nanoparticle chemistry to develop…

  12. Rapid Non-Crosslinking Aggregation of DNA-Functionalized Gold Nanorods and Nanotriangles for Colorimetric Single-Nucleotide Discrimination.

    PubMed

    Wang, Guoqing; Akiyama, Yoshitsugu; Takarada, Tohru; Maeda, Mizuo

    2016-01-01

    Gold nanoparticles modified with DNA duplexes are rapidly and spontaneously aggregated at high ionic strength. In contrast, this aggregation is greatly suppressed when the DNA duplex has a single-base mismatch or a single-nucleotide overhang located at the outermost surface of the particle. These colloidal features emerge irrespective of the size and composition of the particle core; however, the effects of the shape remain unexplored. Using gold nanorods and nanotriangles (nanoplatelets), we show herein that both remarkable rapidity in colloidal aggregation and extreme susceptibility to DNA structural perturbations are preserved, regardless of the shape and aspect ratio of the core. It is also demonstrated that the DNA-modified gold nanorods and nanotriangles are applicable to naked-eye detection of a single-base difference in a gene model. The current study corroborates the generality of the unique colloidal properties of DNA-functionalized nanoparticles, and thus enhances the feasibility of their practical use. PMID:26767586

  13. Simple and rapid quantification of brominated vegetable oil in commercial soft drinks by LC-MS.

    PubMed

    Chitranshi, Priyanka; Gamboa da Costa, Gonçalo

    2016-12-15

    We report here a simple and rapid method for the quantification of brominated vegetable oil (BVO) in soft drinks based upon liquid chromatography-electrospray ionization mass spectrometry. Unlike previously reported methods, this novel method does not require hydrolysis, extraction or derivatization steps, but rather a simple "dilute and shoot" sample preparation. The quantification is conducted by mass spectrometry in selected ion recording mode and a single point standard addition procedure. The method was validated in the range of 5-25μg/mL BVO, encompassing the legal limit of 15μg/mL established by the US FDA for fruit-flavored beverages in the US market. The method was characterized by excellent intra- and inter-assay accuracy (97.3-103.4%) and very low imprecision [0.5-3.6% (RSD)]. The direct nature of the quantification, simplicity, and excellent statistical performance of this methodology constitute clear advantages in relation to previously published methods for the analysis of BVO in soft drinks. PMID:27451219

  14. Colorimetric Method for the Estimation of Total Limonoid Aglycones and Glucoside Contents in Citrus Juices

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A method for estimating the total limonoid aglycone and glucoside concentrations in Citrus samples in terms of limonin and limonin glucoside equivalents is presented. The method consists of extraction followed by colorimetric quantification. The colorimetric quantification was based on the formatio...

  15. Ni(II)NTA AuNPs as a low-resource malarial diagnostic platform for the rapid colorimetric detection of Plasmodium falciparum Histidine-Rich Protein-2.

    PubMed

    Gulka, Christopher P; Swartz, Joshua D; Wright, David W

    2015-04-01

    Diagnosing infectious diseases remains a challenge in the developing world where there is a lack of dependable electricity, running water, and skilled technicians. Although rapid immunochromatographic tests (RDTs) have been deployed to diagnose diseases such as malaria, the extreme climate conditions encountered in these regions compounded with the discrepancies in test manufacturing have yielded varying results, so that more innovative and robust technologies are sought. Devoid of antibodies and thermally sensitive materials, we present a robust, colorimetric diagnostic platform for the detection of a malarial biomarker, Plasmodium falciparum Histidine-Rich Protein 2 (PfHRP-II). The assay exploits the optical properties of gold nanoparticles, covalently coupling them to a Ni(II)NTA recognition element specific for PfHRP-II. In the presence of the recombinant malarial biomarker (rcHRP-II), the Ni(II)NTA AuNPs begin to crosslink and aggregate in as little as one minute, triggering a red-to-purple color change in solution. To increase assay sensitivity and platform stability suitable for low-resource regions, the Ni(II)NTA AuNPs were assembled with varying spacer ligands in a mixed monolayer presentation. When assembled with a negatively charged Peg4-thiol ligand, the Ni(II)NTA AuNPs demonstrate low nanomolar limits of rcHRP-II detection in physiological concentrations of human serum albumin and maintain excellent stability at 37°C when stored for four weeks. Detection of the malaria biomarker is also measured by capturing and processing images of aggregated gold nanoparticles with a smartphone camera. By utilizing a smartphone camera and image processing application, there is no significant difference in assay sensitivity and rcHRP-II limit of detection in comparison to a spectrophotometer, further making this diagnostic platform applicable for use in low-resource regions. PMID:25640131

  16. Simple and Portable Magnetic Immunoassay for Rapid Detection and Sensitive Quantification of Plant Viruses

    PubMed Central

    Rettcher, Stefanie; Jungk, Felicitas; Kühn, Christoph; Krause, Hans-Joachim; Nölke, Greta; Commandeur, Ulrich; Fischer, Rainer; Schillberg, Stefan

    2015-01-01

    Plant pathogens cause major economic losses in the agricultural industry because late detection delays the implementation of measures that can prevent their dissemination. Sensitive and robust procedures for the rapid detection of plant pathogens are therefore required to reduce yield losses and the use of expensive, environmentally damaging chemicals. Here we describe a simple and portable system for the rapid detection of viral pathogens in infected plants based on immunofiltration, subsequent magnetic detection, and the quantification of magnetically labeled virus particles. Grapevine fanleaf virus (GFLV) was chosen as a model pathogen. Monoclonal antibodies recognizing the GFLV capsid protein were immobilized onto immunofiltration columns, and the same antibodies were linked to magnetic nanoparticles. GFLV was quantified by immunofiltration with magnetic labeling in a double-antibody sandwich configuration. A magnetic frequency mixing technique, in which a two-frequency magnetic excitation field was used to induce a sum frequency signal in the resonant detection coil, corresponding to the virus concentration within the immunofiltration column, was used for high-sensitivity quantification. We were able to measure GFLV concentrations in the range of 6 ng/ml to 20 μg/ml in less than 30 min. The magnetic immunoassay could also be adapted to detect other plant viruses, including Potato virus X and Tobacco mosaic virus, with detection limits of 2 to 60 ng/ml. PMID:25710366

  17. Selective colorimetric sensors based on the monitoring of an unmodified silver nanoparticles (AgNPs) reduction for a simple and rapid determination of mercury

    NASA Astrophysics Data System (ADS)

    Jarujamrus, Purim; Amatatongchai, Maliwan; Thima, Araya; Khongrangdee, Thatsanee; Mongkontong, Chakrit

    2015-05-01

    In this work, selective colorimetric sensors for simple and rapid detection of Hg(II) ions based on the monitoring of an unmodified silver nanoparticles (AgNPs) reduction were developed. The average diameter of synthesized AgNPs was 8.3 ± 1.4 nm which was characterized by transmission electron microscopy (TEM). The abrupt change in absorbance of the unmodified AgNPs was observed which progressively decreased and slightly shifted to the blue wavelength as the concentration of Hg(II) increased, indicating the oxidation of Ag(0) to Ag(I) occurred. It appears that the AgNPs were oxidized by Hg(II), resulting in disintegration of the AgNPs into smaller particles as well as mediating the reduction of Hg(II) to Hg(0) adsorbed onto the surface of AgNPs. The adsorption of Hg(0) resulted in the lack of sufficient charges on AgNPs surfaces due to the decrease in the surface coverage of negatively charged citrate molecules, which then leaded to enlargement of AgNPs. The calibration curve of this technique was demonstrated from 0.5 to 7 ppm (r2 = 0.995), the limit of detection (LOD) was 0.06 ppm (SDblank/slope of calibration curve) with the precision (RSD, n = 4) of 3.24-4.53. Interestingly, the results show a significant enhance in the Hg(II) analytical sensitivity when Cu(II) is doped onto the unmodified AgNPs, which improves the quantitative detection limit to 0.008 ppm. In addition, greater selectivity toward Hg(II) compared with the other metal ions tested was observed. Furthermore, the percentage recoveries of spiked drinking water, tap water and SRM1641d (mercury in water) were in acceptable range with a good precision (RSD) which were in agreement with the values obtained from graphite furnace atomic absorption spectrometer (GFAAS). The technique proposed in this study provides a rapid, simple, sensitive and selective detection method for Hg(II) in water samples.

  18. Atomic absorption spectroscopic, conductometric and colorimetric methods for determination of some fluoroquinolone antibacterials using ammonium reineckate

    NASA Astrophysics Data System (ADS)

    Al-Ghannam, Sheikha M.

    2008-04-01

    Three accurate, rapid and simple atomic absorption spectrometric (AAS), conductometric and colorimetric methods were developed for the determination of gatifloxacin (GTF), moxifloxacin (MXF) and sparfloxacin (SPF). The proposed methods depend upon the reaction of ammonium reineckate with the studied drugs to form stable precipitate of ion-pair complexes, which was dissolved in acetone. The pink coloured complexes were determined either by AAS or colorimetrically at λmax 525 nm directly using the dissolved complex. Using conductometric titration, the studied drugs could be evaluated in 50% (v/v) acetone. The optimizations of various experimental conditions were described. Optimum concentration ranges for the determination of GTF, MXF and SPF were 5.0-150, 40-440 μg mL -1 and 0.10-1.5 mg mL -1 using atomic absorption (AAS), conductometric and colorimetric methods, respectively. Detection and quantification limits are ranges from 1.5 to 2.3 μg mL -1 using AAS method or 30-45 μg mL -1 using colorimetric method. The proposed procedures have been applied successfully to the analysis of these drugs in pharmaceutical formulations and the results are favourably comparable to the reference methods.

  19. Novel antibody/gold nanoparticle/magnetic nanoparticle nanocomposites for immunomagnetic separation and rapid colorimetric detection of Staphylococcus aureus in milk.

    PubMed

    Sung, Yun Ju; Suk, Ho-Jun; Sung, Hwa Young; Li, Taihua; Poo, Haryoung; Kim, Min-Gon

    2013-05-15

    We demonstrated the new antibody/gold nanoparticle/magnetic nanoparticle nanocomposites (antibody/AuNP/MNPs) and their application in the detection of the foodborne pathogen, Staphylococcus aureus (S. aureus), in milk. The nanocomposites were synthesized by coating the MNPs with bovine serum albumin (BSA) then adsorbing the AuNPs and anti-S. aureus antibodies on their surface. Using the completed immunomagnetic nanostructures, S. aureus inoculated in the milk sample was captured and isolated from the medium using the permanent magnet. The nanoparticle-bound cells as well as the unbound cells in the supernatant were enumerated via surface plating to evaluate the target binding capacity of the nanocomposites. The capture efficiencies of the antibody/AuNP/MNPs were 96% and 78% for S. aureus in PBS and the milk sample respectively, which were significantly higher than those of the antibody-coupled MNPs without any AuNP. The captured cells were also applied to the selective filtration system to produce color signals that were used for the detection of the target pathogen. During the filtration, the cells bound to the antibody/AuNP/MNPs remained on the surface of the membrane filter while unbound nanoparticles passed through the uniform pores of the membrane. After the gold enhancement, the cells-particles complex resting on the membrane surface rendered a visible color, and the signal intensity became higher as the target cell concentration increased. The detection limits of this colorimetric sensor were 1.5×10(3) and 1.5×10(5)CFU for S. aureus in PBS and the milk sample respectively. This sensing mechanism also had the high specificity for S. aureus over the other pathogens such as Escherichia coli, Listeria monocytogenes, and Salmonella enterica. The assay required only 40min to obtain the results. With the use of the appropriate antibodies, our immunomagnetic nanocomposites-based detection strategy can provide an easy, convenient, and rapid sensing method for a

  20. Toward rapid, high-sensitivity, volume-constrained biomarker quantification and validation using backscattering interferometry.

    PubMed

    Olmsted, Ian R; Hassanein, Mohamed; Kussrow, Amanda; Hoeksema, Megan; Li, Ming; Massion, Pierre P; Bornhop, Darryl J

    2014-08-01

    Realizing personalized medicine, which promises to enable early disease detection, efficient diagnostic staging, and therapeutic efficacy monitoring, hinges on biomarker quantification in patient samples. Yet, the lack of a sensitive technology and assay methodology to rapidly validate biomarker candidates continues to be a bottleneck for clinical translation. In our first direct and quantitative comparison of backscattering interferometry (BSI) to fluorescence sensing by ELISA, we show that BSI could aid in overcoming this limitation. The analytical validation study was performed against ELISA for two biomarkers for lung cancer detection: Cyfra 21-1 and Galectin-7. Spiked serum was used for calibration and comparison of analytical figures of merit, followed by analysis of blinded patient samples. Using the ELISA antibody as the probe chemistry in a mix-and-read assay, BSI provided significantly lower detection limits for spiked serum samples with each of the biomarkers. The limit of quantification (LOQ) for Cyrfa-21-1 was measured to be 230 pg/mL for BSI versus 4000 pg/mL for ELISA, and for Galectin-7, it was 13 pg/mL versus 500 pg/mL. The coefficient of variation for 5 day, triplicate determinations was <15% for BSI and <10% for ELISA. The two techniques correlated well, ranging from 3-29% difference for Cyfra 21-1 in a blinded patient sample analysis. The label-free and free-solution operation of BSI allowed for a significant improvement in analysis speed, with greater ease, improved LOQ values, and excellent day-to-day reproducibility. In this unoptimized format, BSI required 5.5-fold less sample quantity needed for ELISA (a 10 point calibration curve measured in triplicate required 36 μL of serum for BSI vs 200 μL for ELISA). The results indicate that the BSI platform can enable rapid, sensitive analytical validation of serum biomarkers and should significantly impact the validation bottleneck of biomarkers. PMID:24954171

  1. Paper-Based Device for Colorimetric and Photoelectrochemical Quantification of the Flux of H2O2 Releasing from MCF-7 Cancer Cells.

    PubMed

    Li, Li; Zhang, Yan; Zhang, Lina; Ge, Shenguang; Liu, Haiyun; Ren, Na; Yan, Mei; Yu, Jinghua

    2016-05-17

    In this work, a novel dual photoelectrochemical/colorimetric cyto-analysis format was first introduced into a microfluidic paper-based analytical device (μ-PAD) for synchronous sensitive and visual detection of H2O2 released from tumor cells based on an in situ hydroxyl radicals ((•)OH) cleaving DNA approach. The resulted μ-PAD offered an excellent platform for high-performance biosensing applications, which was constructed by a layer-by-layer modification of concanavalin A, graphene quantum dots (GQDs) labeled flower-like Au@Pd alloy nanoparticles (NPs) probe, and tumor cells on the surface of the vertically aligned bamboo like ZnO, which grows on a pyknotic Pt NPs modified paper working electrode (ZnO/Pt-PWE). It was the effective matching of energy levels between GQDs and ZnO levels that lead to the enhancement of the photocurrent response compared with the bare ZnO/Pt-PWE. After releasing H2O2, the DNA strand was cleaved by (•)OH generated under the synergistic catalysis of GQDs and Au@Pd alloy NPs and thus, reduced the photocurrent, resulting in a high sensitivity to H2O2 in aqueous solutions with a detection limit of 0.05 nmol observed, much lower than that in the previously reported method. The disengaged probe can result in catalytic chromogenic reaction of substrates, resulting in real-time imaging of H2O2 biological processes. Therefore, this work provided a truly low-cost, simple, and disposable μ-PAD for precise and visual detection of cellular H2O2, which had potential utility to cellular biology and pathophysiology. PMID:27065042

  2. Rapid detection and quantification of milk adulteration using infrared microspectroscopy and chemometrics analysis.

    PubMed

    Santos, P M; Pereira-Filho, E R; Rodriguez-Saona, L E

    2013-05-01

    The application of attenuated total reflectance mid-infrared microspectroscopy (MIR-microspectroscopy) was evaluated as a rapid method for detection and quantification of milk adulteration. Milk samples were purchased from local grocery stores (Columbus, OH, USA) and spiked at different concentrations of whey, hydrogen peroxide, synthetic urine, urea and synthetic milk. Samples were place on a 192-well microarray slide, air-dried and spectra were collected by using MIR-microspectroscopy. Pattern recognition analysis by Soft Independent Modeling of Class Analogy (SIMCA) showed tight and well-separated clusters allowing discrimination of control samples from adulterated milk. Partial Least Squares Regression (PLSR) showed standard error of prediction (SEP) ~2.33, 0.06, 0.41, 0.30 and 0.014 g/L for estimation of levels of adulteration with whey, synthetic milk, synthetic urine, urea and hydrogen peroxide, respectively. Results showed that MIR-microspectroscopy can provide an alternative methodology to the dairy industry for screening potential fraudulent practice for economic adulteration of cow's milk. PMID:23265450

  3. Rapid quantification of inflammation in tissue samples using perfluorocarbon emulsion and fluorine-19 nuclear magnetic resonance

    PubMed Central

    Ahrens, Eric T.; Young, Won-Bin; Xu, Hongyan; Pusateri, Lisa K.

    2016-01-01

    Quantification of inflammation in tissue samples can be a time-intensive bottleneck in therapeutic discovery and preclinical endeavors. We describe a versatile and rapid approach to quantitatively assay macrophage burden in intact tissue samples. Perfluorocarbon (PFC) emulsion is injected intravenously, and the emulsion droplets are effectively taken up by monocytes and macrophages. These ‘in situ’ labeled cells participate in inflammatory events in vivo resulting in PFC accumulation at inflammatory loci. Necropsied tissues or intact organs are subjected to conventional fluorine-19 (19F) NMR spectroscopy to quantify the total fluorine content per sample, proportional to the macrophage burden. We applied these methods to a rat model of experimental allergic encephalomyelitis (EAE) exhibiting extensive inflammation and demyelination in the central nervous system (CNS), particularly in the spinal cord. In a cohort of EAE rats, we used 19F NMR to derive an inflammation index (IFI) in intact CNS tissues. Immunohistochemistry was used to confirm intracellular colocalization of the PFC droplets within CNS CD68+ cells having macrophage morphology. The IFI linearly correlated to mRNA levels of CD68 via real-time PCR analysis. This 19F NMR approach can accelerate tissue analysis by at least an order of magnitude compared with histological approaches. PMID:21548906

  4. Rapid quantification of live cell receptors using bioluminescence in a flow-based microfluidic device.

    PubMed

    Ramji, Ramesh; Cheong, Cheong Fook; Hirata, Hiroaki; Rahman, Abdur Rub Abdur; Lim, Chwee Teck

    2015-02-25

    The number of receptors expressed by cells plays an important role in controlling cell signaling events, thus determining its behaviour, state and fate. Current methods of quantifying receptors on cells are either laborious or do not maintain the cells in their native form. Here, a method integrating highly sensitive bioluminescence, high precision microfluidics and small footprint of lensfree optics is developed to quantify cell surface receptors. This method is safe to use, less laborious, and faster than the conventional radiolabelling and near field scanning methods. It is also more sensitive than fluorescence based assays and is ideal for high throughput screening. In quantifying β(1) adrenergic receptors expressed on the surface of H9c2 cardiomyocytes, this method yields receptor numbers from 3.12 × 10(5) to 9.36 × 10(5) receptors/cell which are comparable with current methods. This can serve as a very good platform for rapid quantification of receptor numbers in ligand/drug binding and receptor characterization studies, which is an important part of pharmaceutical and biological research. PMID:25336403

  5. Rapid quantification of Salmonella in seafood by real-time PCR assay.

    PubMed

    Kumar, Rakesh; Surendran, P K; Thampuran, Nirmala

    2010-03-01

    A quantitative detection method for Salmonella in seafood was developed using a SYBR Green-based real-time PCR assay. The assay was developed using pure Salmonella DNA at different dilution levels [i.e., 1,000 to 2 genome equivalents (GE)]. The sensitivity of the real-time assay for Salmonella in seeded seafood samples was determined, and the minimum detection level was 20 CFU/g, whereas a detection level of 2 CFU/ml was obtained for pure culture in water with an efficiency of > or =85%. The real-time assay was evaluated in repeated experiments with seeded seafood samples and the regression coefficient (R(2)) values were calculated. The performance of the real-time assay was further assessed with naturally contaminated seafood samples, where 4 out of 9 seafood samples tested positive for Salmonella and harbored cells <100 GE/g, which were not detected by direct plating on Salmonella Chromagar media. Thus, the method developed here will be useful for the rapid quantification of Salmonella in seafood, as the assay can be completed within 2-3 h. In addition, with the ability to detect a low number of Salmonella cells in seafood, this proposed method can be used to generate quantitative data on Salmonella in seafood, facilitating the implementation of control measures for Salmonella contamination in seafood at harvest and post-harvest levels. PMID:20372029

  6. Quantification of microsegregation during rapid solidification of Al-Cu powders

    NASA Astrophysics Data System (ADS)

    Prasad, Arvind; Nenein, Hani; Conlon, Kelly

    2006-05-01

    A new technique is introduced to quantify microsegregation during rapid solidification. The quantification involves calculation of the average solute solubility in the primary phase during solidification of an Al-Cu binary alloy. The calculation is based on using volume percent eutectic and weight percent of second phase (in the eutectic), which were obtained experimentally. Neutron diffraction experiments and stereology calculation on scanning electron microscope images were done on impulse atomized Al-Cu alloys of three compositions (nominal), 5 wt pct Cu, 10 wt pct Cu, and 17 wt pct Cu, atomized under N2 and He gas. Neutron diffraction experiments yielded weight percent CuAl2 data and stereology yielded volume percent eutectic data. These two data were first used to determine the weight percent eutectic. Using the weight percent eutectic and weight percent CuAl2 in mass and volume balance equations, the average solute solubility in the primary phase could be calculated. The experimental results of the amount of eutectic, tomography results from previous work, and results from the calculations suggest that the atomized droplets are in metastable state during the nucleation undercooling of the primary phase, and the effect of metastability propagates through to the eutectic formation stage. The metastable effect is more pronounced in alloys with higher solute composition.

  7. Rapid quantification of microRNAs in plasma using a fast real-time PCR system.

    PubMed

    Andrews, William John; Brown, Eoin Daniel; Dellett, Margaret; Hogg, Ruth Esther; Simpson, David Arthur

    2015-05-01

    The ability to rapidly detect circulating small RNAs, in particular microRNAs (miRNAs), would further increase their already established potential as biomarkers for a range of conditions. One rate-limiting factor in miRNA detection is the time taken to perform quantitative real-time PCR (qPCR) amplification. We therefore evaluated the ability of a novel thermal cycler to perform this step in less than 10 minutes. Quantitative PCR was performed on an xxpress thermal cycler (BJS Biotechnologies), which employs a resistive heating system and forced air cooling to achieve thermal ramp rates of 10°C/s, and a conventional Peltier-controlled LightCycler 480 system (Roche) ramping at 4.8°C/s. The quantification cycle (Cq) for detection of 18S rDNA from a standard genomic DNA sample was significantly more variable across the block (F-test, P = 2.4 × 10(-25)) for the xxpress (20.01 ± 0.47 sd) than for the LightCycler (19.87 ± 0.04 sd). RNA was extracted from human plasma, reverse transcribed, and a panel of miRNAs was amplified and detected using SYBR Green. The sensitivities of the two systems were broadly comparable-both detected a panel of miRNAs reliably, and both indicated similar relative abundances. The xxpress thermal cycler facilitates rapid qPCR detection of small RNAs and brings point-of-care diagnostics based upon detection of circulating miRNAs a step closer to reality. PMID:25967903

  8. Microfluidic Integration of a Cloth-Based Hybridization Array System (CHAS) for Rapid, Colorimetric Detection of Enterohemorrhagic Escherichia coli (EHEC) Using an Articulated, Centrifugal Platform.

    PubMed

    Geissler, Matthias; Clime, Liviu; Hoa, Xuyen D; Morton, Keith J; Hébert, Harold; Poncelet, Lucas; Mounier, Maxence; Deschênes, Mylène; Gauthier, Martine E; Huszczynski, George; Corneau, Nathalie; Blais, Burton W; Veres, Teodor

    2015-10-20

    We describe the translation of a cloth-based hybridization array system (CHAS), a colorimetric DNA detection method that is used by food inspection laboratories for colony screening of pathogenic agents, onto a microfluidic chip format. We also introduce an articulated centrifugal platform with a novel fluid manipulation concept based on changes in the orientation of the chip with respect to the centrifugal force field to time the passage of multiple components required for the process. The platform features two movable and motorized carriers that can be reoriented on demand between 0 and 360° during stage rotation. Articulation of the chip can be used to trigger on-the-fly fluid dispensing through independently addressable siphon structures or to relocate solutions against the centrifugal force field, making them newly accessible for downstream transfer. With the microfluidic CHAS, we achieved significant reduction in the size of the cloth substrate as well as the volume of reagents and wash solutions. Both the chip design and the operational protocol were optimized to perform the entire process in a reliable, fully automated fashion. A demonstration with PCR-amplified genomic DNA confirms on-chip detection and identification of Escherichia coli O157:H7 from colony isolates in a colorimetric multiplex assay using rfbO157, fliCH7, vt1, and vt2 genes. PMID:26416260

  9. Rapid separation and quantification of major caseins and whey proteins of bovine milk by capillary electrophoresis.

    PubMed

    Vallejo-Cordoba, B

    1997-01-01

    A rapid capillary zone electrophoresis (CZE) method was established for separating and quantifying major casein and whey proteins in milk. Optimum sample preparation and electrophoretic conditions in a coated capillary maintained at 40 degrees C allowed accurate and reproducible quantification of milk proteins in a single analysis. Sample and run buffer allowed caseins to be maintained in solution by using a combination of urea and a nonionic detergent in phosphate buffer at pH 2.5. Quantitative CZE protein data were derived by calculating percentages and concentrations (mg/mL) of alpha-casein, beta-casein, alpha-lactalbumin, and beta-lactoglobulin. Calibration curves followed linear relationships with highly significant (p < 0.1) correlation coefficients. Relative standard deviations of less than 0.82 (%) for migration times and 2.18 (%) for percent protein indicated that the technique was reproducible. Electrophoretic protein profiles of fresh bovine milk and rehydrated dry milk showed marked quantitative differences in whey protein concentrations. Whey protein represented 12.37 +/- 0.07% beta-lactoglobulin and 3.05 +/- 0.08% alpha-lactalbumin of total protein in typical fresh milk, while only 1.90 +/- 0.16% beta-lactoglobulin and 0.86 +/- 0.04% alpha-lactalbumin of total protein were detected in a commercial rehydrated milk powder. By quantifying these differences, the established technique may allow the detection of substitution of fresh milk with rehydrated milk powder. The accuracy and reproducibility of the technique permitted the quantitation of individual protein concentrations in milk samples, which agreed with ranges reported in the literature. CZE may be well suited for routine use by dairies and regulatory agencies, since it allows the determination of milk proteins in less than 60 min. PMID:9725120

  10. Ultraminiaturized assay for rapid, low cost detection and quantification of clinical and biochemical samples.

    PubMed

    Parween, Shahila; Nahar, Pradip

    2016-04-01

    Herein, we report a simple, sensitive, rapid and low-cost ultraminiaturized assay technique for quantitative detection of 1 μl of clinical or biochemical sample on a novel ultraminiaturized assay plate (UAP). UAP is prepared by making tiny cavities on a polypropylene sheet. As UAP cannot immobilize a biomolecule through absorption, we have activated the tiny cavities of UAP by 1-fluoro-2-nitro-4-azidobenzene in a photochemical reaction. Activated UAP (AUAP) can covalently immobilize any biomolecule having an active nucleophilic group such as amino group. Efficacy of AUAP is demonstrated by detecting human IgE, antibody of hepatitis C virus core antigen and oligonucleotides. Quantification is performed by capturing the image of the colored assay solution and digitally quantifying the image by color saturation without using costly NanoDrop spectrophotometer. Image - based detection of human IgE and an oligonucleotide shows an excellent correlation with absorbance - based assay (recorded in a NanoDrop spectrophotometer); it is validated by Pearson's product-moment correlation with correlation coefficient of r = 0.9545088 and r = 0.9947444 respectively. AUAP is further checked by detecting hepatitis C virus Ab where strong correlation of color saturation with absorbance with respect to concentration is observed. Ultraminiaturized assay successfully detects target oligonucleotides by perfectly hybridizing with their respective complementary oligonucleotide probes but not with a random oligonucleotide. Ultraminiaturized assay technique has substantially reduced the requirement of reagents by 100 times and assay timing by 50 times making it a potential alternative to conventional method. PMID:26973054

  11. Doped colorimetric assay liposomes

    DOEpatents

    Charych, Deborah; Stevens, Raymond C.

    2001-01-01

    The present invention provides compositions comprising colorimetric assay liposomes. The present invention also provides methods for producing colorimetric liposomes and calorimetric liposome assay systems. In preferred embodiments, these calorimetric liposome systems provide high levels of sensitivity through the use of dopant molecules. As these dopants allow the controlled destabilization of the liposome structure, upon exposure of the doped liposomes to analyte(s) of interest, the indicator color change is facilitated and more easily recognized.

  12. Potential of Visible and Near Infrared Spectroscopy and Pattern Recognition for Rapid Quantification of Notoginseng Powder with Adulterants

    PubMed Central

    Nie, Pengcheng; Wu, Di; Sun, Da-Wen; Cao, Fang; Bao, Yidan; He, Yong

    2013-01-01

    Notoginseng is a classical traditional Chinese medical herb, which is of high economic and medical value. Notoginseng powder (NP) could be easily adulterated with Sophora flavescens powder (SFP) or corn flour (CF), because of their similar tastes and appearances and much lower cost for these adulterants. The objective of this study is to quantify the NP content in adulterated NP by using a rapid and non-destructive visible and near infrared (Vis-NIR) spectroscopy method. Three wavelength ranges of visible spectra, short-wave near infrared spectra (SNIR) and long-wave near infrared spectra (LNIR) were separately used to establish the model based on two calibration methods of partial least square regression (PLSR) and least-squares support vector machines (LS-SVM), respectively. Competitive adaptive reweighted sampling (CARS) was conducted to identify the most important wavelengths/variables that had the greatest influence on the adulterant quantification throughout the whole wavelength range. The CARS-PLSR models based on LNIR were determined as the best models for the quantification of NP adulterated with SFP, CF, and their mixtures, in which the rP values were 0.940, 0.939, and 0.867 for the three models respectively. The research demonstrated the potential of the Vis-NIR spectroscopy technique for the rapid and non-destructive quantification of NP containing adulterants. PMID:24129019

  13. Potential of visible and near infrared spectroscopy and pattern recognition for rapid quantification of notoginseng powder with adulterants.

    PubMed

    Nie, Pengcheng; Wu, Di; Sun, Da-Wen; Cao, Fang; Bao, Yidan; He, Yong

    2013-01-01

    Notoginseng is a classical traditional Chinese medical herb, which is of high economic and medical value. Notoginseng powder (NP) could be easily adulterated with Sophora flavescens powder (SFP) or corn flour (CF), because of their similar tastes and appearances and much lower cost for these adulterants. The objective of this study is to quantify the NP content in adulterated NP by using a rapid and non-destructive visible and near infrared (Vis-NIR) spectroscopy method. Three wavelength ranges of visible spectra, short-wave near infrared spectra (SNIR) and long-wave near infrared spectra (LNIR) were separately used to establish the model based on two calibration methods of partial least square regression (PLSR) and least-squares support vector machines (LS-SVM), respectively. Competitive adaptive reweighted sampling (CARS) was conducted to identify the most important wavelengths/variables that had the greatest influence on the adulterant quantification throughout the whole wavelength range. The CARS-PLSR models based on LNIR were determined as the best models for the quantification of NP adulterated with SFP, CF, and their mixtures, in which the rP values were 0.940, 0.939, and 0.867 for the three models respectively. The research demonstrated the potential of the Vis-NIR spectroscopy technique for the rapid and non-destructive quantification of NP containing adulterants. PMID:24129019

  14. Rapid and in vivo quantification of cellular lipids in Chlorella vulgaris using near-infrared Raman spectrometry.

    PubMed

    Lee, Tsung-Hua; Chang, Jo-Shu; Wang, Hsiang-Yu

    2013-02-19

    A rapid and noninvasive quantification method for cellular lipids in Chlorella vulgaris is demonstrated in this study. This method applied near-infrared Raman spectroscopy to monitor the change of signal intensities at 1440 cm(-1) and 2845-3107 cm(-1) along the nitrogen depletion period, and calibration curves relating signal intensity and cellular lipid abundance were established. The calibration curves show that signal intensity at 2845-3107 cm(-1) and cellular lipid abundance were highly correlated. When the calibration curve was applied on the lipid quantification of two unknown samples, the differences between lipid abundances estimated by the calibration curve and measured by gas chromatography were less than 2 wt %. Carotenoids produced a strong and broad peak near 1440 cm(-1), and it weakened the correlation between signal intensity and lipid abundance. The consistency of detection and effects of cellular contents and water on the Raman spectrogram of Chlorella vulgaris were also addressed. The sample pretreatment only involved centrifugation, and the time required for lipid quantification was shortened to less than 1.5 h. The rapid detection has great potential in high-throughput screening of microalgae and also provides valuable information for monitoring the quality of microalgae culture and determining parameters for the mass production of biodiesel from microalgae. PMID:23331037

  15. RNA-Skim: a rapid method for RNA-Seq quantification at transcript level

    PubMed Central

    Zhang, Zhaojun; Wang, Wei

    2014-01-01

    Motivation: RNA-Seq technique has been demonstrated as a revolutionary means for exploring transcriptome because it provides deep coverage and base pair-level resolution. RNA-Seq quantification is proven to be an efficient alternative to Microarray technique in gene expression study, and it is a critical component in RNA-Seq differential expression analysis. Most existing RNA-Seq quantification tools require the alignments of fragments to either a genome or a transcriptome, entailing a time-consuming and intricate alignment step. To improve the performance of RNA-Seq quantification, an alignment-free method, Sailfish, has been recently proposed to quantify transcript abundances using all k-mers in the transcriptome, demonstrating the feasibility of designing an efficient alignment-free method for transcriptome quantification. Even though Sailfish is substantially faster than alternative alignment-dependent methods such as Cufflinks, using all k-mers in the transcriptome quantification impedes the scalability of the method. Results: We propose a novel RNA-Seq quantification method, RNA-Skim, which partitions the transcriptome into disjoint transcript clusters based on sequence similarity, and introduces the notion of sig-mers, which are a special type of k-mers uniquely associated with each cluster. We demonstrate that the sig-mer counts within a cluster are sufficient for estimating transcript abundances with accuracy comparable with any state-of-the-art method. This enables RNA-Skim to perform transcript quantification on each cluster independently, reducing a complex optimization problem into smaller optimization tasks that can be run in parallel. As a result, RNA-Skim uses <4% of the k-mers and <10% of the CPU time required by Sailfish. It is able to finish transcriptome quantification in <10 min per sample by using just a single thread on a commodity computer, which represents >100 speedup over the state-of-the-art alignment-based methods, while delivering

  16. A high throughput solubility assay for drug discovery using microscale shake-flask and rapid UHPLC-UV-CLND quantification.

    PubMed

    Lin, Baiwei; Pease, Joseph H

    2016-04-15

    The rapid determination of key physical properties of lead compounds is essential to the drug discovery process. Solubility is one of the most important properties since good solubility is needed not only for obtaining reliable in vitro and in vivo assay results in early discovery but also to ensure sufficient concentration of the drug being in circulation to get the desired therapeutic exposure at the target of interest. In order for medicinal chemists to tune solubility of lead compounds, a rapid assay is needed to provide solubility data that is accurate and predictive so that it can be reliably used for designing the next generation of compounds with improved properties. To ensure speed and data quality, we developed a high throughput solubility assay that utilizes a single calibration UHPLC-UV-CLND method and a 24h shake-flask format for rapid quantification. A set of 46 model compounds was used to demonstrate that the method is accurate, reproducible and predictive. Here we present development of the assay, including evaluation of quantification method, filtration membranes, equilibrium times, DMSO concentrations, and buffer conditions. A comparison of thermodynamic solubility results to our high throughput 24h shake-flask solubility assay results is also discussed. PMID:26855285

  17. Rapid testing and quantification of Salmonella in ileocaecal lymph nodes of Austrian pigs slaughtered for consumption.

    PubMed

    Mann, Evelyne; Wagner, Martin; Schmoll, Friedrich; Slaghuis, Jörg; Schönenbrücher, Holger; Mester, Patrick

    2014-10-01

    Traditionally, quantitative microbial risk assessment (QMRA) is based on culture-dependent technologies. However, molecular quantification could forge additional, detailed information. A prerequisite of quantitative real-time PCR in animal science is a tissue preparation method where large volumes of tissue material can be reduced and particularly target cells can be concentrated. An easy-to-use sample preparation method for food (Matrix-Lysis) was recently adapted to tissues and now permits quantification of target cells from up to 5 g of organic matrix. The aim of this study was to examine the suitability of Matrix-Lysis for quantification of Salmonella in porcine ileocaecal lymph nodes (ICLNs). After demonstrating constant recovery rates, ICLNs from 540 pigs were examined for Salmonella spp. with Matrix-Lysis. Samples were also analysed using ISO 6579:2002, a combined enrichment/qPCR method and a lateral flow test. It could be shown that qPCR coupled with Matrix-Lysis can contribute to QMRA in food safety by enabling reproducible quantitative data, even at low contamination rates. PMID:25048837

  18. Rapid Quantification of Methamphetamine: Using Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR) and Chemometrics

    PubMed Central

    Hughes, Juanita; Ayoko, Godwin; Collett, Simon; Golding, Gary

    2013-01-01

    In Australia and increasingly worldwide, methamphetamine is one of the most commonly seized drugs analysed by forensic chemists. The current well-established GC/MS methods used to identify and quantify methamphetamine are lengthy, expensive processes, but often rapid analysis is requested by undercover police leading to an interest in developing this new analytical technique. Ninety six illicit drug seizures containing methamphetamine (0.1%–78.6%) were analysed using Fourier Transform Infrared Spectroscopy with an Attenuated Total Reflectance attachment and Chemometrics. Two Partial Least Squares models were developed, one using the principal Infrared Spectroscopy peaks of methamphetamine and the other a Hierarchical Partial Least Squares model. Both of these models were refined to choose the variables that were most closely associated with the methamphetamine % vector. Both of the models were excellent, with the principal peaks in the Partial Least Squares model having Root Mean Square Error of Prediction 3.8, R2 0.9779 and lower limit of quantification 7% methamphetamine. The Hierarchical Partial Least Squares model had lower limit of quantification 0.3% methamphetamine, Root Mean Square Error of Prediction 5.2 and R2 0.9637. Such models offer rapid and effective methods for screening illicit drug samples to determine the percentage of methamphetamine they contain. PMID:23936058

  19. [Evaluation of Basic Performance of "Point Strip ferritin-3000" for Simple and Rapid Quantification of Serum Ferritin].

    PubMed

    Shibusa, Kotoe; Hatayama, Mayumi; Toki, Yasumichi; Yamamoto, Masayo; Ito, Satoshi; Shindo, Motohiro; Fujiya, Mikihiro; Niizeki, Noriyasu; Tomoda, Yutaka; Kawai, Yuichi; Addo, Lynda; Ikuta, Katsuya

    2015-12-01

    Serum ferritin is an excellent marker for total iron content in the body and is essential for the diagnosis of iron deficiency or iron overload. Recently, a simple and rapid method, which utilizes immunochromatography for the quantification of serum ferritin, was developed. However, the range of measurement in previous reagents was limited (10-500 ng/mL). This range is rather narrow and is not fully helpful for the diagnosis of iron overload which sometimes occurs as a result of prolonged transfusions, or for monitoring iron contents during iron chelation therapy against iron overload. In the present study we evaluated the basic performance of the newly developed "Point Strip ferritin-3000", which can measure serum ferritin in the range of 300-3,000 ng/mL. Coefficient of variation (CV) s of within and inter-day assays were in the ranges of 7.3-11.1% and 2.1-5.2%, respectively. Using 87 serum samples obtained from the patients with written informed consents, the correlation coefficient was calculated to be 0.93 compared to the control method. In addition, the quantification of serum ferritin by "Point Strip ferritin-3000" was not influenced by bilirubin, hemoglobin, chyle, rheumatoid factor, or ascorbic acid. From our data, "Point Strip ferritin-3000" is reliable reagent in the range of 300-3,000 ng/mL, and is therefore considered to be useful for the diagnosis of iron overload, as well as for monitoring iron contents during iron chelation therapy. In addition, this quantification method can be easily performed using a small desktop equipment without any special technique, making this system applicable for epidemiological surveys and clinical studies. PMID:27089653

  20. Rapid turbidimetric assay for quantification of fusidic acid in a dermatological cream.

    PubMed

    Curbete, Mariane Machado; Salgado, Hérida Regina Nunes

    2016-06-01

    Fusidic acid is an antibiotic steroid widely used for the treatment of serious infections caused by methicillin-resistant Staphylococcus aureus (MRSA) strains. Microbiological methods are indispensable to determine the mean percentage of antimicrobial in medicaments during manufacturing and quality control processes. The aim of this study was to develop and validate a microbiological method for the quantification of fusidic acid in dermatological cream by turbidimetry, using Staphylococcus epidermidis (ATCC 12228) and casoy broth as the culture medium. The validation parameters were in accordance with ICH specifications and demonstrated accuracy, precision, selectivity, and robustness, with linear ranges from 0.25 to 2.25μgmL(-1). This method is an alternative to the diffusion agar assay currently employed to quantify fusidic acid in dermatological cream, since it is sensitive, fast, and more economical. PMID:27130089

  1. Rapid quantification of low level polymorph content in a solid dose form using transmission Raman spectroscopy.

    PubMed

    Griffen, Julia A; Owen, Andrew W; Burley, Jonathan; Taresco, Vincenzo; Matousek, Pavel

    2016-09-01

    This proof of concept study demonstrates the application of transmission Raman spectroscopy (TRS) to the non-invasive and non-destructive quantification of low levels (0.62-1.32% w/w) of an active pharmaceutical ingredient's polymorphic forms in a pharmaceutical formulation. Partial least squares calibration models were validated with independent validation samples resulting in prediction RMSEP values of 0.03-0.05% w/w and a limit of detection of 0.1-0.2% w/w. The study further demonstrates the ability of TRS to quantify all tablet constituents in one single measurement. By analysis of degraded stability samples, sole transformation between polymorphic forms was observed while excipient levels remained constant. Additionally, a beam enhancer device was used to enhance laser coupling to the sample, which allowed comparable prediction performance at 60 times faster rates (0.2s) than in standard mode. PMID:27218440

  2. Quantification of Primary Dendritic and Secondary Eutectic Nucleation Undercoolings in Rapidly Solidified Hypo-Eutectic Al-Cu Droplets

    NASA Astrophysics Data System (ADS)

    Bogno, A.-A.; Khatibi, P. Delshad; Henein, H.; Gandin, Ch.-A.

    2016-09-01

    This paper reports on the quantification of primary dendritic and secondary eutectic nucleation undercoolings during rapid solidification of impulse atomized hypo-eutectic Al-Cu droplets. The procedure consists in determining the eutectic fraction of each investigated droplet from the fraction of intermetallic Al2Cu obtained by Rietveld refinement analysis of neutrons scattering data. The corresponding eutectic nucleation undercooling is then deduced from the metastable phase diagram of the alloy. The primary dendritic nucleation undercooling is subsequently determined using semi-empirical coarsening models of secondary dendrite arms. The two nucleation undercoolings are finally used as input variables to run a microsegregation model for binary alloys. The fractions of eutectic computed by the microsegregation model compare very favorably with the experimental results.

  3. Quantification of Primary Dendritic and Secondary Eutectic Nucleation Undercoolings in Rapidly Solidified Hypo-Eutectic Al-Cu Droplets

    NASA Astrophysics Data System (ADS)

    Bogno, A.-A.; Khatibi, P. Delshad; Henein, H.; Gandin, Ch.-A.

    2016-06-01

    This paper reports on the quantification of primary dendritic and secondary eutectic nucleation undercoolings during rapid solidification of impulse atomized hypo-eutectic Al-Cu droplets. The procedure consists in determining the eutectic fraction of each investigated droplet from the fraction of intermetallic Al2Cu obtained by Rietveld refinement analysis of neutrons scattering data. The corresponding eutectic nucleation undercooling is then deduced from the metastable phase diagram of the alloy. The primary dendritic nucleation undercooling is subsequently determined using semi-empirical coarsening models of secondary dendrite arms. The two nucleation undercoolings are finally used as input variables to run a microsegregation model for binary alloys. The fractions of eutectic computed by the microsegregation model compare very favorably with the experimental results.

  4. Development of a real-time PCR assay for rapid detection and quantification of Alexandrium minutum (a Dinoflagellate).

    PubMed

    Galluzzi, Luca; Penna, Antonella; Bertozzini, Elena; Vila, Magda; Garcés, Esther; Magnani, Mauro

    2004-02-01

    The marine dinoflagellate genus Alexandrium includes a number of species which produce neurotoxins responsible for paralytic shellfish poisoning (PSP), which in humans may cause muscular paralysis, neurological symptoms, and, in extreme cases, death. A. minutum is the most widespread toxic PSP species in the western Mediterranean basin. The monitoring of coastal waters for the presence of harmful algae also normally involves microscopic examinations of phytoplankton populations. These procedures are time consuming and require a great deal of taxonomic experience, thus limiting the number of specimens that can be analyzed. Because of the genetic diversity of different genera and species, molecular tools may also help to detect the presence of target microorganisms in marine field samples. In this study, we developed a real-time PCR-based assay for rapid detection of all toxic species of the Alexandrium genus in both fixative-preserved environmental samples and cultures. Moreover, we developed a real-time quantitative PCR assay for the quantification of A. minutum cells in seawater samples. Alexandrium genus-specific primers were designed on the 5.8S rDNA region. Primer specificity was confirmed by using BLAST and by amplification of a representative sample of the DNA of other dinoflagellates and diatoms. Using a standard curve constructed with a plasmid containing the ITS1-5.8S-ITS2 A. minutum sequence and cultured A. minutum cells, we determined the absolute number of 5.8S rDNA copies per cell. Consequently, after quantification of 5.8S rDNA copies in samples containing A. minutum cells, we were also able to estimate the number of cells. Several fixed A. minutum bloom sea samples from Arenys Harbor (Catalan Coast, Spain) were analyzed using this method, and quantification results were compared with standard microscopy counting methods. The two methods gave comparable results, confirming that real-time PCR could be a valid, fast alternative procedure for the

  5. A Rapid Molecular Test for Determining Yersinia pestis Susceptibility to Ciprofloxacin by the Quantification of Differentially Expressed Marker Genes

    PubMed Central

    Steinberger-Levy, Ida; Shifman, Ohad; Zvi, Anat; Ariel, Naomi; Beth-Din, Adi; Israeli, Ofir; Gur, David; Aftalion, Moshe; Maoz, Sharon; Ber, Raphael

    2016-01-01

    Standard antimicrobial susceptibility tests used to determine bacterial susceptibility to antibiotics are growth dependent and time consuming. The long incubation time required for standard tests may render susceptibility results irrelevant, particularly for patients infected with lethal bacteria that are slow growing on agar but progress rapidly in vivo, such as Yersinia pestis. Here, we present an alternative approach for the rapid determination of antimicrobial susceptibility, based on the quantification of the changes in the expression levels of specific marker genes following exposure to growth-inhibiting concentrations of the antibiotic, using Y. pestis and ciprofloxacin as a model. The marker genes were identified by transcriptomic DNA microarray analysis of the virulent Y. pestis Kimberley53 strain after exposure to specific concentrations of ciprofloxacin for various time periods. We identified several marker genes that were induced following exposure to growth-inhibitory concentrations of ciprofloxacin, and we confirmed the marker expression profiles at additional ciprofloxacin concentrations using quantitative RT-PCR. Eleven candidate marker transcripts were identified, of which four mRNA markers were selected for a rapid quantitative RT-PCR susceptibility test that correctly determined the Minimal Inhibitory Concentration (MIC) values and the categories of susceptibility of several Y. pestis strains and isolates harboring various ciprofloxacin MIC values. The novel molecular susceptibility test requires just 2 h of antibiotic exposure in a 7-h overall test time, in contrast to the 24 h of antibiotic exposure required for a standard microdilution test. PMID:27242774

  6. A Rapid Molecular Test for Determining Yersinia pestis Susceptibility to Ciprofloxacin by the Quantification of Differentially Expressed Marker Genes.

    PubMed

    Steinberger-Levy, Ida; Shifman, Ohad; Zvi, Anat; Ariel, Naomi; Beth-Din, Adi; Israeli, Ofir; Gur, David; Aftalion, Moshe; Maoz, Sharon; Ber, Raphael

    2016-01-01

    Standard antimicrobial susceptibility tests used to determine bacterial susceptibility to antibiotics are growth dependent and time consuming. The long incubation time required for standard tests may render susceptibility results irrelevant, particularly for patients infected with lethal bacteria that are slow growing on agar but progress rapidly in vivo, such as Yersinia pestis. Here, we present an alternative approach for the rapid determination of antimicrobial susceptibility, based on the quantification of the changes in the expression levels of specific marker genes following exposure to growth-inhibiting concentrations of the antibiotic, using Y. pestis and ciprofloxacin as a model. The marker genes were identified by transcriptomic DNA microarray analysis of the virulent Y. pestis Kimberley53 strain after exposure to specific concentrations of ciprofloxacin for various time periods. We identified several marker genes that were induced following exposure to growth-inhibitory concentrations of ciprofloxacin, and we confirmed the marker expression profiles at additional ciprofloxacin concentrations using quantitative RT-PCR. Eleven candidate marker transcripts were identified, of which four mRNA markers were selected for a rapid quantitative RT-PCR susceptibility test that correctly determined the Minimal Inhibitory Concentration (MIC) values and the categories of susceptibility of several Y. pestis strains and isolates harboring various ciprofloxacin MIC values. The novel molecular susceptibility test requires just 2 h of antibiotic exposure in a 7-h overall test time, in contrast to the 24 h of antibiotic exposure required for a standard microdilution test. PMID:27242774

  7. Fractal circuit sensors enable rapid quantification of biomarkers for donor lung assessment for transplantation

    PubMed Central

    Sage, Andrew T.; Besant, Justin D.; Mahmoudian, Laili; Poudineh, Mahla; Bai, Xiaohui; Zamel, Ricardo; Hsin, Michael; Sargent, Edward H.; Cypel, Marcelo; Liu, Mingyao; Keshavjee, Shaf; Kelley, Shana O.

    2015-01-01

    Biomarker profiling is being rapidly incorporated in many areas of modern medical practice to improve the precision of clinical decision-making. This potential improvement, however, has not been transferred to the practice of organ assessment and transplantation because previously developed gene-profiling techniques require an extended period of time to perform, making them unsuitable in the time-sensitive organ assessment process. We sought to develop a novel class of chip-based sensors that would enable rapid analysis of tissue levels of preimplantation mRNA markers that correlate with the development of primary graft dysfunction (PGD) in recipients after transplant. Using fractal circuit sensors (FraCS), three-dimensional metal structures with large surface areas, we were able to rapidly (<20 min) and reproducibly quantify small differences in the expression of interleukin-6 (IL-6), IL-10, and ATP11B mRNA in donor lung biopsies. A proof-of-concept study using 52 human donor lungs was performed to develop a model that was used to predict, with excellent sensitivity (74%) and specificity (91%), the incidence of PGD for a donor lung. Thus, the FraCS-based approach delivers a key predictive value test that could be applied to enhance transplant patient outcomes. This work provides an important step toward bringing rapid diagnostic mRNA profiling to clinical application in lung transplantation. PMID:26601233

  8. New protocol for the rapid quantification of exopolysaccharides in continuous culture systems of acidophilic bioleaching bacteria.

    PubMed

    Michel, Caroline; Bény, Claire; Delorme, Fabian; Poirier, Laurence; Spolaore, Pauline; Morin, Dominique; d'Hugues, Patrick

    2009-02-01

    In this study, we investigate exopolysaccharide production by a bacterial consortium during the bioleaching of a cobaltiferrous pyrite. Whereas comparable studies have looked at exopolysaccharide production in batch systems, this study focuses on a continuous system comprising a series of four stirred bioreactors and reveals the difficulties in quantifying biomolecules in complex media such as bioleached samples. We also adapted the phenol/sulphuric acid method to take into account iron interference, thus establishing a new protocol for sugar quantification in bioleached samples characterised by low pH (1.4) and high iron concentration (2 g l(-1)). This allows sugar analysis without any prior sample preparation step; only a small amount of sample is needed (0.5 ml) and sample preparation is limited to a single filtration step. We found that free exopolysaccharides represented more than 80% of the total sugars in the bioreactors, probably because stirring creates abrasive conditions and detaches sugars bound to pyrite or bacteria and that they were produced mainly in the first two reactors where bioleaching activity was greatest. However, we could not establish any direct link between the measured exopolysaccharide concentration and bioleaching activity. Exopolysaccharides could have another role (protection against stress) in addition to that in bacterial attachment. PMID:19130051

  9. Development of an assay for rapid detection and quantification of Verticillium dahliae in soil.

    PubMed

    Bilodeau, Guillaume J; Koike, Steven T; Uribe, Pedro; Martin, Frank N

    2012-03-01

    ABSTRACT Verticillium dahliae is responsible for Verticillium wilt on a wide range of hosts, including strawberry, on which low soil inoculum densities can cause significant crop loss. Determination of inoculum density is currently done by soil plating but this can take 6 to 8 weeks to complete and delay the grower's ability to make planting decisions. To provide a faster means for estimating pathogen populations in the soil, a multiplexed TaqMan real-time polymerase chain reaction (PCR) assay based on the ribosomal DNA (rDNA) intergenic spacer (IGS) was developed for V. dahliae. The assay was specific for V. dahliae and included an internal control for evaluation of inhibition due to the presence of PCR inhibitors in DNA extracted from soil samples. An excellent correlation was observed in regression analysis (R(2) = 0.96) between real-time PCR results and inoculum densities determined by soil plating in a range of field soils with pathogen densities as low as 1 to 2 microsclerotia/g of soil. Variation in copy number of the rDNA was also evaluated among isolates by SYBR Green real-time PCR amplification of the V. dahliae-specific amplicon compared with amplification of several single-copy genes and was estimated to range from ≈24 to 73 copies per haploid genome, which translated into possible differences in results among isolates of ≈1.8 cycle thresholds. Analysis of the variation in results of V. dahliae quantification among extractions of the same soil sample indicated that assaying four replicate DNA extractions for each field sample would provide accurate results. A TaqMan assay also was developed to help identify colonies of V. tricorpus on soil plates. PMID:22066673

  10. Rapid Quantification of Hepatitis B Virus DNA by Automated Sample Preparation and Real-Time PCR

    PubMed Central

    Stelzl, Evelyn; Muller, Zsofia; Marth, Egon; Kessler, Harald H.

    2004-01-01

    Monitoring of hepatitis B virus (HBV) DNA in serum by molecular methods has become the standard for assessment of the replicative activity of HBV. Several molecular assays for the detection and quantification of HBV DNA have been described. However, they usually lack automated sample preparation. Moreover, those assays, which are based on PCR, are limited by a short dynamic range (2 to 3 log units). In the present study, the use of RealArt HBV LC PCR Reagents in conjunction with automated extraction on the COBAS AMPLIPREP analyzer was evaluated. Members of an HBV proficiency program panel were tested; linearity, interassay, and intra-assay variations were determined. The performance of the assay in a routine clinical laboratory was evaluated with a total of 117 clinical specimens. When members of the HBV proficiency program panel were tested by the new molecular assay, the results were found to be within ±0.5 log unit of the results obtained by reference laboratories. Determination of linearity resulted in a quasilinear curve over more than 6 log units. The interassay variation of the RealArt HBV LC PCR Reagents by use of the automated sample preparation protocol ranged from 16 to 73%, and the intra-assay variation ranged from 9 to 40%. When clinical samples were tested by the new assay with the automated sample preparation protocol and the results were compared with those obtained by the COBAS AMPLICOR HBV MONITOR Test with manual sample preparation, the results for 76% of all samples with positive results by both tests were found to be within ±0.5 log unit and the results for another 18% were found to be within between 0.5 and 1.0 log unit. In conclusion, the real-time PCR assay with automated sample preparation proved to be suitable for the routine molecular laboratory and required less hands-on time. PMID:15184417

  11. Rapid imaging, detection and quantification of Giardia lamblia cysts using mobile-phone based fluorescent microscopy and machine learning.

    PubMed

    Koydemir, Hatice Ceylan; Gorocs, Zoltan; Tseng, Derek; Cortazar, Bingen; Feng, Steve; Chan, Raymond Yan Lok; Burbano, Jordi; McLeod, Euan; Ozcan, Aydogan

    2015-03-01

    Rapid and sensitive detection of waterborne pathogens in drinkable and recreational water sources is crucial for treating and preventing the spread of water related diseases, especially in resource-limited settings. Here we present a field-portable and cost-effective platform for detection and quantification of Giardia lamblia cysts, one of the most common waterborne parasites, which has a thick cell wall that makes it resistant to most water disinfection techniques including chlorination. The platform consists of a smartphone coupled with an opto-mechanical attachment weighing ~205 g, which utilizes a hand-held fluorescence microscope design aligned with the camera unit of the smartphone to image custom-designed disposable water sample cassettes. Each sample cassette is composed of absorbent pads and mechanical filter membranes; a membrane with 8 μm pore size is used as a porous spacing layer to prevent the backflow of particles to the upper membrane, while the top membrane with 5 μm pore size is used to capture the individual Giardia cysts that are fluorescently labeled. A fluorescence image of the filter surface (field-of-view: ~0.8 cm(2)) is captured and wirelessly transmitted via the mobile-phone to our servers for rapid processing using a machine learning algorithm that is trained on statistical features of Giardia cysts to automatically detect and count the cysts captured on the membrane. The results are then transmitted back to the mobile-phone in less than 2 minutes and are displayed through a smart application running on the phone. This mobile platform, along with our custom-developed sample preparation protocol, enables analysis of large volumes of water (e.g., 10-20 mL) for automated detection and enumeration of Giardia cysts in ~1 hour, including all the steps of sample preparation and analysis. We evaluated the performance of this approach using flow-cytometer-enumerated Giardia-contaminated water samples, demonstrating an average cyst capture

  12. Rapid onsite detection of bacterial spores of biothreat importance by paper-based colorimetric method using erbium-pyrocatechol violet complex.

    PubMed

    Shivakiran, M S; Venkataramana, M; Lakshmana Rao, P V

    2016-01-01

    Dipicolinic acid (DPA) is an important chemical marker for the detection of bacterial spores. In this study, complexes of lanthanide series elements such as erbium, europium, neodymium, and terbium were prepared with pyrocatechol violet and effectively immobilized the pyrocatechol violet (PV)-metal complex on a filter paper using polyvinyl alcohol. These filter paper strips were employed for the onsite detection of bacterial spores. The test filter papers were evaluated quantitatively with different concentrations of DPA and spores of various bacteria. Among the four lanthanide ions, erbium displayed better sensitivity than the other ions. The limit of detection of this test for DPA was 60 μM and 5 × 10(6) spores. The effect of other non-spore-forming bacteria and interfering chemicals on the test strips was also evaluated. The non-spore-forming bacteria did not have considerable effect on the test strip whereas chemicals such as EDTA had significant effects on the test results. The present test is rapid and robust, capable of providing timely results for better judgement to save resources on unnecessary decontamination procedures during false alarms. PMID:26603759

  13. Specific ionic effect for simple and rapid colorimetric sensing assays of amino acids using gold nanoparticles modified with task-specific ionic liquid.

    PubMed

    Wu, Datong; Cai, Pengfei; Tao, Zhihao; Pan, Yuanjiang

    2016-01-01

    In this study, a novel task-specific ionic liquid functionalized gold nanoparticle (TSIL-GNP) was successfully prepared and applied in the recognition of amino acids. Particularly, the surface of GNP was modified with the ionic liquid containing carbamido and ester group via thiol, which was characterized by Fourier transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM). The stability of this material in aqueous solution improves apparently and can remain unchanged for more than three months. The effect of pH was also discussed in this study. Attractive ionic interaction would effectively weaken intensity of the covalent coupling between the metal ion and the functional groups of amino acids. Thus, TSIL-GNP was successfully applied to recognizing serine, aspartic acid, lysine, arginine, and histidine in the presence of Cu(2+) through distinctive color changes. Suspension would be generated once a spot of cysteine was added into the GNPs solution. Results indicated that it had a good linear relationship between extinction coefficients and concentration of amino acids in a wide range of 10(-3)-10(-6) M. Moreover, the proposed strategy was successfully used to analyze the histidine in urinary samples. In brief, TSIL-GNP is a suitable substrate for discrimination of five amino acids in a rapid and simple way without sophisticated instruments. PMID:26703268

  14. Quantification of anthropogenic and natural changes in oil sands mining infrastructure land based on RapidEye and SPOT5

    NASA Astrophysics Data System (ADS)

    Zhang, Ying; Guindon, Bert; Lantz, Nicholas; Shipman, Todd; Chao, Dennis; Raymond, Don

    2014-06-01

    Natural resources development, spanning exploration, production and transportation activities, alters local land surface at various spatial scales. Quantification of these anthropogenic changes, both permanent and reversible, is needed for compliance assessment and for development of effective sustainable management strategies. Multi-spectral high resolution imagery data from SPOT5 and RapidEye were used for extraction and quantification of the anthropogenic and natural changes for a case study of Alberta bitumen (oil sands) mining located in the Western Boreal Plains near Fort McMurray, Canada. Two test sites representative of the major Alberta bitumen production extraction processes, open pit and in situ extraction, were selected. A hybrid change detection approach, combining pixel- and object-based target detection and extraction, is proposed based on Change Vector Analysis (CVA). The extraction results indicate that the changed infrastructure landscapes of these two sites have different footprints linked with their differing oil sands production processes. Pixel- and object-based accuracy assessments have been applied for validation of the change detection results. For manmade disturbances, except for those fine linear features such as the seismic lines, accuracies of about 80% have been achieved at the pixel level while, at the object level, these rise to 90-95%. Since many disturbance features are transient, a new landscape index, entitled the Re-growth Index, has been formulated at single object level specifically to monitor restoration of these features to their natural state. It is found that the temporal behaviour of the Re-growth Index in an individual patch varies depending on the type of natural land cover. In addition, the Re-growth Index is also useful for assessing the detectability of disturbed sites.

  15. Semiautomatic and rapid quantification of heartbeat parameters in Drosophila using optical coherence tomography imaging

    NASA Astrophysics Data System (ADS)

    Guo, Shou-Yuan; Liao, Fang-Tsu; Su, Ming-Tsan; Chang, Cheng-Yi; Su, Hong-Ren; Huang, Jyun-Cin; Kuo, Wen-Chuan

    2013-02-01

    We report a semiautomatic algorithm that is specialized for rapid analysis of beat-to-beat contraction-relaxation parameters of the heart in Drosophila. The presented algorithm adapts the general graph theoretical image segmentation algorithm and a histogram-based thresholding algorithm, which can measure many cardiac parameters, including heart rate, heart period, diastolic and systolic intervals, and end-diastolic and end-systolic areas. Additionally, dynamic cardiac functions, such as arrhythmia index and percent fractional shortening, can be automatically calculated for all the recorded heartbeats over significant periods of time.

  16. Rapid Quantification of 3D Collagen Fiber Alignment and Fiber Intersection Correlations with High Sensitivity

    PubMed Central

    Sun, Meng; Bloom, Alexander B.; Zaman, Muhammad H.

    2015-01-01

    Metastatic cancers aggressively reorganize collagen in their microenvironment. For example, radially orientated collagen fibers have been observed surrounding tumor cell clusters in vivo. The degree of fiber alignment, as a consequence of this remodeling, has often been difficult to quantify. In this paper, we present an easy to implement algorithm for accurate detection of collagen fiber orientation in a rapid pixel-wise manner. This algorithm quantifies the alignment of both computer generated and actual collagen fiber networks of varying degrees of alignment within 5°°. We also present an alternative easy method to calculate the alignment index directly from the standard deviation of fiber orientation. Using this quantitative method for determining collagen alignment, we demonstrate that the number of collagen fiber intersections has a negative correlation with the degree of fiber alignment. This decrease in intersections of aligned fibers could explain why cells move more rapidly along aligned fibers than unaligned fibers, as previously reported. Overall, our paper provides an easier, more quantitative and quicker way to quantify fiber orientation and alignment, and presents a platform in studying effects of matrix and cellular properties on fiber alignment in complex 3D environments. PMID:26158674

  17. Rapid Quantification of 3D Collagen Fiber Alignment and Fiber Intersection Correlations with High Sensitivity.

    PubMed

    Sun, Meng; Bloom, Alexander B; Zaman, Muhammad H

    2015-01-01

    Metastatic cancers aggressively reorganize collagen in their microenvironment. For example, radially orientated collagen fibers have been observed surrounding tumor cell clusters in vivo. The degree of fiber alignment, as a consequence of this remodeling, has often been difficult to quantify. In this paper, we present an easy to implement algorithm for accurate detection of collagen fiber orientation in a rapid pixel-wise manner. This algorithm quantifies the alignment of both computer generated and actual collagen fiber networks of varying degrees of alignment within 5°°. We also present an alternative easy method to calculate the alignment index directly from the standard deviation of fiber orientation. Using this quantitative method for determining collagen alignment, we demonstrate that the number of collagen fiber intersections has a negative correlation with the degree of fiber alignment. This decrease in intersections of aligned fibers could explain why cells move more rapidly along aligned fibers than unaligned fibers, as previously reported. Overall, our paper provides an easier, more quantitative and quicker way to quantify fiber orientation and alignment, and presents a platform in studying effects of matrix and cellular properties on fiber alignment in complex 3D environments. PMID:26158674

  18. Device for rapid quantification of human carotid baroreceptor-cardiac reflex responses

    NASA Technical Reports Server (NTRS)

    Sprenkle, J. M.; Eckberg, D. L.; Goble, R. L.; Schelhorn, J. J.; Halliday, H. C.

    1986-01-01

    A new device has been designed, constructed, and evaluated to characterize the human carotid baroreceptor-cardiac reflex response relation rapidly. This system was designed for study of reflex responses of astronauts before, during, and after space travel. The system comprises a new tightly sealing silicon rubber neck chamber, a stepping motor-driven electrodeposited nickel bellows pressure system, capable of delivering sequential R-wave-triggered neck chamber pressure changes between +40 and -65 mmHg, and a microprocessor-based electronics system for control of pressure steps and analysis and display of responses. This new system provokes classic sigmoid baroreceptor-cardiac reflex responses with threshold, linear, and saturation ranges in most human volunteers during one held expiration.

  19. TREND: a tool for rapid online research literature analysis and quantification.

    PubMed

    Landers, Richard N

    2008-08-01

    The Research Explicator for oNline Databases (TREND) tool was developed out of a need to quantify large research literatures rapidly and objectively on the basis of online research database output. By parsing such output with TREND, a researcher can in minutes extract the most commonly cited articles, the most frequently published authors, a distribution of publication dates, and a variety of other information from a research literature several thousand articles in size. This tool thus enables an increase in productivity both for researchers venturing into new areas of interest and for advisors and instructors putting together core reading lists. The processing of citations from articles represents a unique challenge, however, because deviations from strict APA formatting cause problems that are sometimes difficult to correct mechanically. A case study of one particularly troublesome citation (Baron & Kenny, 1986) is presented. Usage and implications are discussed. PMID:18697661

  20. Quantification of cell viability and rapid screening anti-cancer drug utilizing nanomechanical fluctuation.

    PubMed

    Wu, Shangquan; Liu, Xiaoli; Zhou, Xiarong; Liang, Xin M; Gao, Dayong; Liu, Hong; Zhao, Gang; Zhang, Qingchuan; Wu, Xiaoping

    2016-03-15

    Cancer is a serious threat to human health. Although numerous anti-cancer drugs are available clinically, many have shown toxic side effects due to poor tumor-selectivity, and reduced effectiveness due to cancers rapid development of resistance to treatment. The development of new highly efficient and practical methods to quantify cell viability and its change under drug treatment is thus of significant importance in both understanding of anti-cancer mechanism and anti-cancer drug screening. Here, we present an approach of utilizing a nanomechanical fluctuation based highly sensitive microcantilever sensor, which is capable of characterizing the viability of cells and quantitatively screening (within tens of minutes) their responses to a drug with the obvious advantages of a rapid, label-free, quantitative, noninvasive, real-time and in-situ assay. The microcantilever sensor operated in fluctuation mode was used in evaluating the paclitaxel effectiveness on breast cancer cell line MCF-7. This study demonstrated that the nanomechanical fluctuations of the microcantilever sensor are sensitive enough to detect the dynamic variation in cellular force which is provided by the cytoskeleton, using cell metabolism as its energy source, and the dynamic instability of microtubules plays an important role in the generation of the force. We propose that cell viability consists of two parts: biological viability and mechanical viability. Our experimental results suggest that paclitaxel has little effect on biological viability, but has a significant effect on mechanical viability. This new method provides a new concept and strategy for the evaluation of cell viability and the screening of anti-cancer drugs. PMID:26406457

  1. Rapid and simple UPLC-MS/MS method for precise phytochelatin quantification in alga extracts.

    PubMed

    Bräutigam, Anja; Wesenberg, Dirk; Preud'homme, Hugues; Schaumlöffel, Dirk

    2010-09-01

    Quantitative phytochelatin (PC) analysis is, due to oxidation sensitivity of the PCs, matrix effects, and time consuming sample preparation, still a challenging analytical task. In this study, a rapid, simple, and sensitive method for precise determination of native PCs in crude extracts of the green alga Chlamydomonas reinhardtii was developed. Algae were exposed 48 h to 70 μM Cd. Coupling of ultra performance liquid chromatography and electrospray ionization tandem mass spectrometry with multi-reaction mode transitions for detection permitted the required short-time, high-resolution separation and detection specificity. Thus, under optimized chromatographic conditions, 10 thiol peptides were baseline-separated within 7 min. Relative detection limits in the nanomolar range in microliter sample volumes were achieved (corresponding to absolute detection limits at femtomole level). Next to glutathione (GSH), the most abundant cadmium-induced PCs in C. reinhardtii, namely CysGSH, PC(2), PC(3), CysPC(2), and CysPC(3), were quantified with high reproducibility at concentrations between 15 and 198 nmol g(-1) fresh weight. The biological variation of PC synthesis of nine independently grown alga cultures was determined to be on average 13.7%. PMID:20632163

  2. Rapid guided wave delamination detection and quantification in composites using global-local sensing

    NASA Astrophysics Data System (ADS)

    Tian, Zhenhua; Yu, Lingyu; Leckey, Cara

    2016-08-01

    This paper presents a rapid guided ultrasonic wave inspection approach through global inspection by phased array beamforming and local damage evaluation via wavenumber analysis. The global-local approach uses a hybrid system consisting of a PZT wafer and a non-contact laser vibrometer. The overall inspection is performed in two steps. First, a phased array configured by a small number of measurements performs beamforming and beamsteering over the entire plate in order to detect and locate the presence of the damage. A local area is identified as target damage area for the second step. Then a high density wavefield measurement is taken over the target damage area and a spatial wavenumber imaging is performed to quantitatively evaluate the damage. The two-step inspection has been applied to locate and quantify impact-induced delamination damage in a carbon fiber reinforced polymer composite plate. The detected delamination location, size and shape agree well with those of an ultrasonic C-scan. For the test case studied in this work the global-local approach reduced the total composite inspection (damage detection and characterization) time by ∼97% compared to using a full scan approach.

  3. Electromechanical transducer for rapid detection, discrimination and quantification of lung cancer cells

    NASA Astrophysics Data System (ADS)

    Ali, Waqas; Jalvhei Moghaddam, Fatemeh; Usman Raza, Muhammad; Bui, Loan; Sayles, Bailey; Kim, Young-Tae; Iqbal, Samir M.

    2016-05-01

    Tumor cells are malignant derivatives of normal cells. There are characteristic differences in the mechanophysical properties of normal and tumor cells, and these differences stem from the changes that occur in the cell cytoskeleton during cancer progression. There is a need for viable whole blood processing techniques for rapid and reliable tumor cell detection that do not require tagging. Micropore biosensors have previously been used to differentiate tumor cells from normal cells and we have used a micropore-based electromechanical transducer to differentiate one type of tumor cells from the other types. This device generated electrical signals that were characteristic of the cell properties. Three non-small cell lung cancer (NSCLC) cell lines, NCl-H1155, A549 and NCI-H460, were successfully differentiated. NCI-H1155, due to their comparatively smaller size, were found to be the quickest in translocating through the micropore. Their translocation through a 15 μm micropore caused electrical pulses with an average translocation time of 101 ± 9.4 μs and an average peak amplitude of 3.71 ± 0.42 μA, whereas translocation of A549 and NCI-H460 caused pulses with average translocation times of 126 ± 17.9 μs and 148 ± 13.7 μs and average peak amplitudes of 4.58 ± 0.61 μA and 5.27 ± 0.66 μA, respectively. This transformation of the differences in cell properties into differences in the electrical profiles (i.e. the differences in peak amplitudes and translocation times) with this electromechanical transducer is a quantitative way to differentiate these lung cancer cells. The solid-state micropore device processed whole biological samples without any pre-processing requirements and is thus ideal for point-of-care applications.

  4. Sensitive and rapid RT-qPCR quantification of pathogenic Candida species in human blood.

    PubMed

    Ogata, Kiyohito; Matsuda, Kazunori; Tsuji, Hirokazu; Nomoto, Koji

    2015-10-01

    For accurate diagnosis and appropriate treatment of candidiasis, we developed a highly sensitive quantitative RT-PCR (RT-qPCR) system for five Candida species that have been reported to be the major causes of bloodstream fungal infection (Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis, and Candida krusei), together with a system for all pathogenic Candida species. Cells of each fungal species spiked into human peripheral blood (PB) were specifically detected at a lower detection limit of 10(0) cell/1 mL PB by this system using the newly developed specific primer sets targeting 18S or 26S rRNA of the five Candida species, together with the existing group primer set. The total count of the five Candida spp. as the sum of those obtained by using the five species primer sets was equivalent to the count obtained by using the group primer set, indicating that the group set covered the major five Candida spp. in human blood with the same degree of accuracy as the species primer sets. The RT-qPCR counts of the Candida species were in good agreement with CFU counts obtained by their culture on CHROMagar™, with a lower detection limit of 10(0)cell/mL of PB. Candida rRNA molecules were stably stored for at least 7 days at 4°C by keeping the blood specimens in an RNA stabilizing reagent. These results strongly suggest that this sensitive system is useful for accurate and rapid diagnosis of Candida bloodstream infections. PMID:26232708

  5. Rapid TaqMan-Based Quantification of Chlorophyll d-Containing Cyanobacteria in the Genus Acaryochloris

    PubMed Central

    Behrendt, Lars; Nielsen, Jeppe L.; Sørensen, Søren J.; Larkum, Anthony W. D.; Winther, Jakob R.

    2014-01-01

    Reports of the chlorophyll (Chl) d-containing cyanobacterium Acaryochloris have accumulated since its initial discovery in 1996. The majority of this evidence is based on amplification of the gene coding for the 16S rRNA, and due to the wide geographical distribution of these sequences, a global distribution of Acaryochloris species was suggested. Here, we present a rapid, reliable, and cost-effective TaqMan-based quantitative PCR (qPCR) assay that was developed for the specific detection of Acaryochloris species in complex environmental samples. The TaqMan probe showed detection limits of ∼10 16S rRNA gene copy numbers based on standard curves consisting of plasmid inserts. DNA from five Acaryochloris strains, i.e., MBIC11017, CCMEE5410, HICR111A, CRS, and Awaji-1, exhibited amplification efficiencies of >94% when tested in the TaqMan assay. When used on complex natural communities, the TaqMan assay detected the presence of Acaryochloris species in four out of eight samples of crustose coralline algae (CCA), collected from temperate and tropical regions. In three out of these TaqMan-positive samples, the presence of Chl d was confirmed via high-performance liquid chromatography (HPLC), and corresponding cell estimates of Acaryochloris species amounted to 7.6 × 101 to 3.0 × 103 per mg of CCA. These numbers indicate a substantial contribution of Chl d-containing cyanobacteria to primary productivity in endolithic niches. The new TaqMan assay allows quick and easy screening of environmental samples for the presence of Acaryochloris species and is an important tool to further resolve the global distribution and significance of this unique oxyphototroph. PMID:24632258

  6. Rapid extraction, identification and quantification of oral hypoglycaemic drugs in serum and hair using LC-MS/MS.

    PubMed

    Binz, Tina M; Villani, Nicholas; Neels, Hugo; Schneider, Serge

    2012-11-30

    A sensitive and accurate LC-MS/MS method for the identification and quantification of 5 oral anti-diabetics (glipizide, glibenclamide, gliclazide, gliquidone and metformin) in serum and hair was developed using glibornuride as the internal standard. We have developed a rapid and robust extraction procedure by using acetonitrile for serum protein precipitation and methanol for the extraction of anti-diabetics from hair. Anti-diabetics (ADs) were separated by UPLC over a C18 column and detection was performed on a Waters Xevo TQ MS mass spectrometer in positive ionization mode using electrospray ionization. Each AD was identified by three specific ion transitions in multiple reaction monitoring (MRM) mode. The method was validated according to international guidelines. For all compounds the variation coefficient (CV) was <20%, and accuracies ranged from 85 to 115% in serum and hair. The limits of detection (LODs) were <1.5 ng/mL for all ADs in serum and <3.59 pg/mg in hair. Recoveries varied from 56.41% (gliclazide) to 67.58% (glipizide) in serum and from 68% (gliclazide) to 91.2% (metformin) in hair. The method was successfully applied to quantify ADs in serum of 33 patients and in hair of 15 patients. PMID:22940189

  7. Rapid method for quantification of seven synthetic pigments in colored Chinese steamed buns using UFLC-MS/MS without SPE.

    PubMed

    Gao, He-Gang; Gong, Wen-Jie; Zhao, Yong-Gang

    2015-01-01

    Synthetic pigments are still used instead of natural pigments in many foods and their residues in food could be an important risk to human health. A simple and rapid analytical method combining the low-cost extraction protocol with ultra-fast liquid chromatography-tandem quadrupole mass spectrometry (UFLC-MS/MS) was developed for the simultaneous determination of seven synthetic pigments used in colored Chinese steamed buns. For the first time, ethanol/ammonia solution/water (7:2:1, v/v/v) was used as extraction solution for the synthetic pigments in colored Chinese steamed buns. The results showed that the property of the extraction solution used in this method was more effective than critic acid solution, which is used in the polyamide adsorption method. The limits of quantification for the seven synthetic pigments ranged from 0.15 to 0.50 μg/kg. The present method was successfully applied to samples of colored Chinese steamed buns for food-safety risk monitoring in Zhejiang Province, China. The results found sunset yellow pigment in six out of 300 colored Chinese steamed buns (from 0.50 to 32.6 μg/kg). PMID:25765275

  8. Electromechanical transducer for rapid detection, discrimination and quantification of lung cancer cells.

    PubMed

    Ali, Waqas; Moghaddam, Fatemeh Jalvhei; Raza, Muhammad Usman; Bui, Loan; Sayles, Bailey; Kim, Young-Tae; Iqbal, Samir M

    2016-05-13

    Tumor cells are malignant derivatives of normal cells. There are characteristic differences in the mechanophysical properties of normal and tumor cells, and these differences stem from the changes that occur in the cell cytoskeleton during cancer progression. There is a need for viable whole blood processing techniques for rapid and reliable tumor cell detection that do not require tagging. Micropore biosensors have previously been used to differentiate tumor cells from normal cells and we have used a micropore-based electromechanical transducer to differentiate one type of tumor cells from the other types. This device generated electrical signals that were characteristic of the cell properties. Three non-small cell lung cancer (NSCLC) cell lines, NCl-H1155, A549 and NCI-H460, were successfully differentiated. NCI-H1155, due to their comparatively smaller size, were found to be the quickest in translocating through the micropore. Their translocation through a 15 μm micropore caused electrical pulses with an average translocation time of 101 ± 9.4 μs and an average peak amplitude of 3.71 ± 0.42 μA, whereas translocation of A549 and NCI-H460 caused pulses with average translocation times of 126 ± 17.9 μs and 148 ± 13.7 μs and average peak amplitudes of 4.58 ± 0.61 μA and 5.27 ± 0.66 μA, respectively. This transformation of the differences in cell properties into differences in the electrical profiles (i.e. the differences in peak amplitudes and translocation times) with this electromechanical transducer is a quantitative way to differentiate these lung cancer cells. The solid-state micropore device processed whole biological samples without any pre-processing requirements and is thus ideal for point-of-care applications. PMID:27023745

  9. Development of a colorimetric sensor array for squid spoilage assessment.

    PubMed

    Zaragozá, Patricia; Fuentes, Ana; Ruiz-Rico, María; Vivancos, José-Luis; Fernández-Segovia, Isabel; Ros-Lis, José V; Barat, José M; Martínez-Máñez, Ramón

    2015-05-15

    The aim of this work was to develop and evaluate a rapid, easy-to-use optoelectronic system for the shelf-life assessment of squid in cold storage. For this purpose, an optoelectronic nose was designed, which consisted of an array containing six sensing materials prepared by combining different dyes and two inorganic supports (aluminium oxide and silica gel). Samples were packaged with the colorimetric array and kept in cold storage for 12 days. Squid spoilage was monitored simultaneously by the colorimetric array and by the physico-chemical and microbial analyses during storage. Samples exceeded the acceptability limits for microbial counts on the third day. PCA analysis carried out with CIELab showed that the colorimetric array was able to discriminate between fresh squid fit for consumption and spoiled squid. The statistical models obtained by PLS, with the optoelectronic nose, successfully predicted CO2 and O2 content in the headspace as well as microbial growth. PMID:25577086

  10. Rapid, simple and highly sensitive LC-ESI-MS/MS method for the quantification of tamsulosin in human plasma.

    PubMed

    Ramakrishna, N V S; Vishwottam, K N; Manoj, S; Koteshwara, M; Wishu, S; Varma, D P

    2005-12-01

    A simple, rapid, sensitive and specific liquid chromatography-tandem mass spectrometry method was developed and validated for quantification of tamsulosin (I), a highly selective alpha1-adrenoceptor antagonist used for the treatment of patients with symptomatic benign prostatic hyperplasia. The analyte and internal standard, mosapride (II) were extracted by liquid-liquid extraction with diethyl ether-dichloromethane (70:30, v/v) using a Glas-Col Multi-Pulse Vortexer. The chromatographic separation was performed on a reverse phase Waters symmetry C18 column with a mobile phase of 0.03% formic acid-acetonitrile (30:70, v/v). The protonated analyte was quantitated in positive ionization by multiple reaction monitoring with a mass spectrometer. The mass transitions m/z 409.1 solidus in circle 228.1 and m/z 422.3 solidus in circle 198.3 were used to measure I and II, respectively. The assay exhibited a linear dynamic range of 0.1-50.0 ng/mL for tamsulosin in human plasma. The lower limit of quantitation was 100 pg/mL with a relative standard deviation of less than 10%. Acceptable precision and accuracy were obtained for concentrations over the standard curve ranges. A run time of 2.0 min for each sample made it possible to analyze a throughput of more than 400 human plasma samples per day. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetic, bioavailability or bioequivalence studies. PMID:15828055

  11. Quantification of rapid environmental redox processes with quick-scanning x-ray absorption spectroscopy (Q-XAS).

    PubMed

    Ginder-Vogel, Matthew; Landrot, Gautier; Fischel, Jason S; Sparks, Donald L

    2009-09-22

    Quantification of the initial rates of environmental reactions at the mineral/water interface is a fundamental prerequisite to determining reaction mechanisms and contaminant transport modeling and predicting environmental risk. Until recently, experimental techniques with adequate time resolution and elemental sensitivity to measure initial rates of the wide variety of environmental reactions were quite limited. Techniques such as electron paramagnetic resonance and Fourier transform infrared spectroscopies suffer from limited elemental specificity and poor sensitivity to inorganic elements, respectively. Ex situ analysis of batch and stirred-flow systems provides high elemental sensitivity; however, their time resolution is inadequate to characterize rapid environmental reactions. Here we apply quick-scanning x-ray absorption spectroscopy (Q-XAS), at sub-second time-scales, to measure the initial oxidation rate of As(III) to As(V) by hydrous manganese(IV) oxide. Using Q-XAS, As(III) and As(V) concentrations were determined every 0.98 s in batch reactions. The initial apparent As(III) depletion rate constants (t < 30 s) measured with Q-XAS are nearly twice as large as rate constants measured with traditional analytical techniques. Our results demonstrate the importance of developing analytical techniques capable of analyzing environmental reactions on the same time scale as they occur. Given the high sensitivity, elemental specificity, and time resolution of Q-XAS, it has many potential applications. They could include measuring not only redox reactions but also dissolution/precipitation reactions, such as the formation and/or reductive dissolution of Fe(III) (hydr)oxides, solid-phase transformations (i.e., formation of layered-double hydroxide minerals), or almost any other reaction occurring in aqueous media that can be measured using x-ray absorption spectroscopy. PMID:19805269

  12. Individual Colorimetric Observer Model

    PubMed Central

    Asano, Yuta; Fairchild, Mark D.; Blondé, Laurent

    2016-01-01

    This study proposes a vision model for individual colorimetric observers. The proposed model can be beneficial in many color-critical applications such as color grading and soft proofing to assess ranges of color matches instead of a single average match. We extended the CIE 2006 physiological observer by adding eight additional physiological parameters to model individual color-normal observers. These eight parameters control lens pigment density, macular pigment density, optical densities of L-, M-, and S-cone photopigments, and λmax shifts of L-, M-, and S-cone photopigments. By identifying the variability of each physiological parameter, the model can simulate color matching functions among color-normal populations using Monte Carlo simulation. The variabilities of the eight parameters were identified through two steps. In the first step, extensive reviews of past studies were performed for each of the eight physiological parameters. In the second step, the obtained variabilities were scaled to fit a color matching dataset. The model was validated using three different datasets: traditional color matching, applied color matching, and Rayleigh matches. PMID:26862905

  13. A versatile targeted metabolomics method for the rapid quantification of multiple classes of phenolics in fruits and beverages.

    PubMed

    Vrhovsek, Urska; Masuero, Domenico; Gasperotti, Mattia; Franceschi, Pietro; Caputi, Lorenzo; Viola, Roberto; Mattivi, Fulvio

    2012-09-12

    Compelling evidence of the health benefits of phenolic compounds and their impact on food quality have stimulated the development of analytical methods for the identification and quantification of these compounds in different matrices in recent years. A targeted metabolomics method has been developed for the quantification of 135 phenolics, such as benzoates, phenylpropanoids, coumarins, stilbenes, dihydrochalcones, and flavonoids, in fruit and tea extracts and wine using UPLC/QqQ-MS/MS. Chromatography was optimized to achieve separation of the compounds over a period of 15 min, and MRM transitions were selected for accurate quantification. The method was validated by studying the detection and quantification limits, the linearity ranges, and the intraday and interday repeatability of the analysis. The validated method was applied to the analysis of apples, berries, green tea, and red wine, providing a valuable tool for food quality evaluation and breeding studies. PMID:22468648

  14. A rapid and accurate quantification method for real-time dynamic analysis of cellular lipids during microalgal fermentation processes in Chlorella protothecoides with low field nuclear magnetic resonance.

    PubMed

    Wang, Tao; Liu, Tingting; Wang, Zejian; Tian, Xiwei; Yang, Yi; Guo, Meijin; Chu, Ju; Zhuang, Yingping

    2016-05-01

    The rapid and real-time lipid determination can provide valuable information on process regulation and optimization in the algal lipid mass production. In this study, a rapid, accurate and precise quantification method of in vivo cellular lipids of Chlorella protothecoides using low field nuclear magnetic resonance (LF-NMR) was newly developed. LF-NMR was extremely sensitive to the algal lipids with the limits of the detection (LOD) of 0.0026g and 0.32g/L in dry lipid samples and algal broth, respectively, as well as limits of quantification (LOQ) of 0.0093g and 1.18g/L. Moreover, the LF-NMR signal was specifically proportional to the cellular lipids of C. protothecoides, thus the superior regression curves existing in a wide detection range from 0.02 to 0.42g for dry lipids and from 1.12 to 8.97gL(-1) of lipid concentration for in vivo lipid quantification were obtained with all R(2) higher than 0.99, irrespective of the lipid content and fatty acids profile variations. The accuracy of this novel method was further verified to be reliable by comparing lipid quantification results to those obtained by GC-MS. And the relative standard deviation (RSD) of LF-NMR results were smaller than 2%, suggesting the precision of this method. Finally, this method was successfully used in the on-line lipid monitoring during the algal lipid fermentation processes, making it possible for better understanding of the lipid accumulation mechanism and dynamic bioprocess control. PMID:26948045

  15. Smartphone spectrometer for colorimetric biosensing.

    PubMed

    Wang, Yi; Liu, Xiaohu; Chen, Peng; Tran, Nhung Thi; Zhang, Jinling; Chia, Wei Sheng; Boujday, Souhir; Liedberg, Bo

    2016-05-23

    We report on a smartphone spectrometer for colorimetric biosensing applications. The spectrometer relies on a sample cell with an integrated grating substrate, and the smartphone's built-in light-emitting diode flash and camera. The feasibility of the smartphone spectrometer is demonstrated for detection of glucose and human cardiac troponin I, the latter in conjunction with peptide-functionalized gold nanoparticles. PMID:27163736

  16. Rapid Quantification of Melamine in Different Brands/Types of Milk Powders Using Standard Addition Net Analyte Signal and Near-Infrared Spectroscopy.

    PubMed

    Tang, Bang-Cheng; Cai, Chen-Bo; Shi, Wei; Xu, Lu

    2016-01-01

    Multivariate calibration (MVC) and near-infrared (NIR) spectroscopy have demonstrated potential for rapid analysis of melamine in various dairy products. However, the practical application of ordinary MVC can be largely restricted because the prediction of a new sample from an uncalibrated batch would be subject to a significant bias due to matrix effect. In this study, the feasibility of using NIR spectroscopy and the standard addition (SA) net analyte signal (NAS) method (SANAS) for rapid quantification of melamine in different brands/types of milk powders was investigated. In SANAS, the NAS vector of melamine in an unknown sample as well as in a series of samples added with melamine standards was calculated and then the Euclidean norms of series standards were used to build a straightforward univariate regression model. The analysis results of 10 different brands/types of milk powders with melamine levels 0~0.12% (w/w) indicate that SANAS obtained accurate results with the root mean squared error of prediction (RMSEP) values ranging from 0.0012 to 0.0029. An additional advantage of NAS is to visualize and control the possible unwanted variations during standard addition. The proposed method will provide a practically useful tool for rapid and nondestructive quantification of melamine in different brands/types of milk powders. PMID:27525154

  17. Rapid Quantification of Melamine in Different Brands/Types of Milk Powders Using Standard Addition Net Analyte Signal and Near-Infrared Spectroscopy

    PubMed Central

    2016-01-01

    Multivariate calibration (MVC) and near-infrared (NIR) spectroscopy have demonstrated potential for rapid analysis of melamine in various dairy products. However, the practical application of ordinary MVC can be largely restricted because the prediction of a new sample from an uncalibrated batch would be subject to a significant bias due to matrix effect. In this study, the feasibility of using NIR spectroscopy and the standard addition (SA) net analyte signal (NAS) method (SANAS) for rapid quantification of melamine in different brands/types of milk powders was investigated. In SANAS, the NAS vector of melamine in an unknown sample as well as in a series of samples added with melamine standards was calculated and then the Euclidean norms of series standards were used to build a straightforward univariate regression model. The analysis results of 10 different brands/types of milk powders with melamine levels 0~0.12% (w/w) indicate that SANAS obtained accurate results with the root mean squared error of prediction (RMSEP) values ranging from 0.0012 to 0.0029. An additional advantage of NAS is to visualize and control the possible unwanted variations during standard addition. The proposed method will provide a practically useful tool for rapid and nondestructive quantification of melamine in different brands/types of milk powders. PMID:27525154

  18. SynPAnal: Software for Rapid Quantification of the Density and Intensity of Protein Puncta from Fluorescence Microscopy Images of Neurons

    PubMed Central

    Danielson, Eric; Lee, Sang H.

    2014-01-01

    Continuous modification of the protein composition at synapses is a driving force for the plastic changes of synaptic strength, and provides the fundamental molecular mechanism of synaptic plasticity and information storage in the brain. Studying synaptic protein turnover is not only important for understanding learning and memory, but also has direct implication for understanding pathological conditions like aging, neurodegenerative diseases, and psychiatric disorders. Proteins involved in synaptic transmission and synaptic plasticity are typically concentrated at synapses of neurons and thus appear as puncta (clusters) in immunofluorescence microscopy images. Quantitative measurement of the changes in puncta density, intensity, and sizes of specific proteins provide valuable information on their function in synaptic transmission, circuit development, synaptic plasticity, and synaptopathy. Unfortunately, puncta quantification is very labor intensive and time consuming. In this article, we describe a software tool designed for the rapid semi-automatic detection and quantification of synaptic protein puncta from 2D immunofluorescence images generated by confocal laser scanning microscopy. The software, dubbed as SynPAnal (for Synaptic Puncta Analysis), streamlines data quantification for puncta density and average intensity, thereby increases data analysis throughput compared to a manual method. SynPAnal is stand-alone software written using the JAVA programming language, and thus is portable and platform-free. PMID:25531531

  19. SynPAnal: software for rapid quantification of the density and intensity of protein puncta from fluorescence microscopy images of neurons.

    PubMed

    Danielson, Eric; Lee, Sang H

    2014-01-01

    Continuous modification of the protein composition at synapses is a driving force for the plastic changes of synaptic strength, and provides the fundamental molecular mechanism of synaptic plasticity and information storage in the brain. Studying synaptic protein turnover is not only important for understanding learning and memory, but also has direct implication for understanding pathological conditions like aging, neurodegenerative diseases, and psychiatric disorders. Proteins involved in synaptic transmission and synaptic plasticity are typically concentrated at synapses of neurons and thus appear as puncta (clusters) in immunofluorescence microscopy images. Quantitative measurement of the changes in puncta density, intensity, and sizes of specific proteins provide valuable information on their function in synaptic transmission, circuit development, synaptic plasticity, and synaptopathy. Unfortunately, puncta quantification is very labor intensive and time consuming. In this article, we describe a software tool designed for the rapid semi-automatic detection and quantification of synaptic protein puncta from 2D immunofluorescence images generated by confocal laser scanning microscopy. The software, dubbed as SynPAnal (for Synaptic Puncta Analysis), streamlines data quantification for puncta density and average intensity, thereby increases data analysis throughput compared to a manual method. SynPAnal is stand-alone software written using the JAVA programming language, and thus is portable and platform-free. PMID:25531531

  20. A rapid Fourier-transform infrared (FTIR) spectroscopic method for direct quantification of paracetamol content in solid pharmaceutical formulations

    NASA Astrophysics Data System (ADS)

    Mallah, Muhammad Ali; Sherazi, Syed Tufail Hussain; Bhanger, Muhammad Iqbal; Mahesar, Sarfaraz Ahmed; Bajeer, Muhammad Ashraf

    2015-04-01

    A transmission FTIR spectroscopic method was developed for direct, inexpensive and fast quantification of paracetamol content in solid pharmaceutical formulations. In this method paracetamol content is directly analyzed without solvent extraction. KBr pellets were formulated for the acquisition of FTIR spectra in transmission mode. Two chemometric models: simple Beer's law and partial least squares employed over the spectral region of 1800-1000 cm-1 for quantification of paracetamol content had a regression coefficient of (R2) of 0.999. The limits of detection and quantification using FTIR spectroscopy were 0.005 mg g-1 and 0.018 mg g-1, respectively. Study for interference was also done to check effect of the excipients. There was no significant interference from the sample matrix. The results obviously showed the sensitivity of transmission FTIR spectroscopic method for pharmaceutical analysis. This method is green in the sense that it does not require large volumes of hazardous solvents or long run times and avoids prior sample preparation.

  1. Quantification of whey proteins by reversed phase-HPLC and effectiveness of mid-infrared spectroscopy for their rapid prediction in sweet whey.

    PubMed

    Sturaro, Alba; De Marchi, Massimo; Masi, Antonio; Cassandro, Martino

    2016-01-01

    In the dairy industry, membrane filtration is used to reduce the amount of whey waste and, simultaneously, to recover whey proteins (WP). The composition of WP can strongly affect the filtration treatment of whey, and rapid determination of WP fractions would be of interest for dairy producers to monitor WP recovery. This study aimed to develop mid-infrared spectroscopy (MIRS) prediction models for the rapid quantification of protein in sweet whey, using a validated rapid reversed phase (RP)-HPLC as a reference method. Quantified WP included α-lactalbumin (α-LA), β-lactoglobulin (β-LG) A and B, bovine serum albumin, caseinomacropeptides, and proteose peptone. Validation of RP-HPLC was performed by calculating the relative standard deviation (RSD) in repeatability and reproducibility tests for WP retention time and peak areas. Samples of liquid whey (n=187) were analyzed by RP-HPLC and scanned through MIRS to collect spectral information (900 to 4,000 cm(-1)); statistical analysis was carried out through partial least squares regression and random cross-validation procedure. Retention times in RP-HPLC method were stable (RSD between 0.03 and 0.80%), whereas the RSD of peak area (from 0.25 to 8.48%) was affected by WP relative abundance. Higher coefficients of determination in validation for MIRS model were obtained for protein fractions present in whey in large amounts, such as β-LG (0.58), total identified WP (0.58), and α-LA (0.56). Results of this study suggest that MIRS is an easy method for rapid quantification of detail protein in sweet whey, even if better resolution was achieved with the method based on RP-HPLC. The prediction of WP in sweet whey by MIRS might be used for screening and for classifying sweet whey according to its total and individual WP contents. PMID:26585472

  2. A colorimetric sensor array of porous pigments.

    PubMed

    Lim, Sung H; Kemling, Jonathan W; Feng, Liang; Suslick, Kenneth S

    2009-12-01

    The development of a low-cost, simple colorimetric sensor array capable of the detection and identification of toxic gases is reported. This technology uses a disposable printed array of porous pigments in which metalloporphyrins and chemically-responsive dyes are immobilized in a porous matrix of organically modified siloxanes (ormosils) and printed on a porous membrane. The printing of the ormosil into the membrane is highly uniform and does not lessen the porosity of the membrane, as shown by scanning electron microscopy. When exposed to an analyte, these pigments undergo reactions that result in well-defined color changes due to strong chemical interactions: ligation to metal ions, Lewis or Brønsted acid-base interactions, hydrogen bonding, etc. Striking visual identification of 3 toxic gases has been shown at the IDLH (immediately dangerous to life and health) concentration, at the PEL (permissible exposure level), and at a level well below the PEL. Identification and quantification of analytes were achieved using the color change profiles, which were readily distinguishable in a hierarchical clustering analysis (HCA) dendrogram, with no misclassifications in 50 trials. PMID:19918616

  3. A colorimetric sensor array of porous pigments

    PubMed Central

    Lim, Sung H.; Kemling, Jonathan W.; Feng, Liang

    2010-01-01

    The development of a low-cost, simple colorimetric sensor array capable of detection and identification of toxic gases is reported. This technology uses a disposable printed array of porous pigments in which metalloporphyrins and chemically responsive dyes are immobilized in a porous matrix of organically modified siloxanes (ormosils) and printed on a porous membrane. The printing of the ormosil into the membrane is highly uniform and does not lessen the porosity of the membrane, as shown by scanning electron microscopy. When exposed to an analyte, these pigments undergo reactions that result in well-defined color changes due to strong chemical interactions: ligation to metal ions, Lewis or Bronsted acid-base interactions, hydrogen bonding, etc. Striking visual identification of 3 toxic gases has been shown at the IDLH (immediately dangerous to life and health), at the PEL (permissible exposure level), and at a level well below the PEL. Identification and quantification of analytes were achieved using the color change profiles, which were readily distinguishable in a hierarchical clustering analysis (HCA) dendrogram, with no misclassifications in 50 trials. PMID:19918616

  4. Colorimetric characterization of LED luminaires

    NASA Astrophysics Data System (ADS)

    Costa, C. L. M.; Vieira, R. R.; Pereira, R. C.; Silva, P. V. M.; Oliveira, I. A. A.; Sardinha, A. S.; Viana, D. D.; Barbosa, A. H.; Souza, L. P.; Alvarenga, A. D.

    2015-01-01

    The Optical Metrology Division of Inmetro - National Institute of Metrology, Quality and Technology has recently started the colorimetric characterization of lamps by implementing Correlated Color Temperature (CCT) and Color Rendering Index (CRI) measurements of incandescent lamps, followed by the CFL, and LED lamps and luminaires. Here we present the results for the verification of the color characterization of samples of SSL luminaires for public as well as indoor illumination that are sold in Brazil.

  5. Rapid quantification of major reaction products formed during thermochemical pretreatment of lignocellulosic biomass using GC-MS.

    PubMed

    Humpula, James F; Chundawat, Shishir P S; Vismeh, Ramin; Jones, A Daniel; Balan, Venkatesh; Dale, Bruce E

    2011-04-15

    Accurate quantification of reaction products formed during thermochemical pretreatment of lignocellulosic biomass would lead to a better understanding of plant cell wall deconstruction for production of cellulosic biofuels and biochemicals. However, quantification of some process byproducts, most notably acetamide, acetic acid and furfural, present several analytical challenges using conventional liquid chromatography methods. Therefore, we have developed a high-throughput gas chromatography based mass spectrometric (GC-MS) method in order to quantify relevant compounds without requiring time-consuming sample derivatization prior to analysis. Solvent extracts of untreated, ammonia fiber expansion (AFEX) treated and dilute-acid treated corn stover were analyzed by this method. Biomass samples were extracted with acetone using an automated solvent extractor, serially diluted and directly analyzed using the proposed GC-MS method. Acetone was the only solvent amongst water, methanol and acetonitrile that did not contain detectable background levels of the target compounds or facilitate a buildup of plant-derived residues in the GC injector, which decreased analytical reproducibility. Quantitative results were based on the method of standard addition and external standard calibration curves. PMID:21444255

  6. Rapid and sensitive quantification of C3- and C6-phosphoesters in starch by fluorescence-assisted capillary electrophoresis.

    PubMed

    Verbeke, Jeremy; Penverne, Christophe; D'Hulst, Christophe; Rolando, Christian; Szydlowski, Nicolas

    2016-11-01

    Phosphate groups are naturally present in starch at C3- or C6-position of the glucose residues and impact the structure of starch granules. Their precise quantification is necessary for understanding starch physicochemical properties and metabolism. Nevertheless, reliable quantification of Glc-3-P remains laborious and time consuming. Here we describe a capillary electrophoresis method for simultaneous measurement of both Glc-6-P and Glc-3-P after acid hydrolysis of starch. The sensitivity threshold was estimated at the fg scale, which is compatible with the analysis of less than a μg of sample. The method was validated by analyzing antisense potato lines deficient in SBEs, GWD or GBSS. We show that Glc-3-P content is altered in the latter and that these variations do not correlate with modifications in Glc-6-P content. We anticipate the method reported here to be an efficient tool for high throughput study of starch phosphorylation at both C3- and C6-position. PMID:27516330

  7. Development of a SYBR Green quantitative polymerase chain reaction assay for rapid detection and quantification of infectious laryngotracheitis virus.

    PubMed

    Mahmoudian, Alireza; Kirkpatrick, Naomi C; Coppo, Mauricio; Lee, Sang-Won; Devlin, Joanne M; Markham, Philip F; Browning, Glenn F; Noormohammadi, Amir H

    2011-06-01

    Infectious laryngotracheitis is an acute viral respiratory disease of chickens with a worldwide distribution. Sensitive detection of the causative herpesvirus is particularly important because it can persist in the host at a very low copy number and be transmitted to other birds. Quantification of viral genome copy number is also useful for clinical investigations and experimental studies. In the study presented here, a quantitative polymerase chain reaction (qPCR) assay was developed using SYBR Green chemistry and the viral gene UL15a to detect and quantify infectious laryngotracheitis virus (ILTV) in ILTV-inoculated chicken embryos or naturally infected birds. The specificity of the assay was confirmed using a panel of viral and bacterial pathogens of poultry. The sensitivity of the assay was compared with two conventional PCR assays, virus titration and an antigen-detecting enzyme-linked immunosorbent assay. The qPCR developed in this study was highly sensitive and specific, and has potential for quantification of ILTV in tissues from naturally and experimentally infected birds and embryos. PMID:21711182

  8. Rapid and Thiol-Specific High-Throughput Assay for Simultaneous Relative Quantification of Total Thiols, Protein Thiols, and Nonprotein Thiols in Cells

    PubMed Central

    2015-01-01

    Thiol groups in biological molecules play a significant role in various physiological functions and pathological conditions. Thiols are divided into two major groups: protein thiols and nonprotein thiols. Numerous methods have been reported for thiol assays. Most of these methods have been developed for glutathione, the principal nonprotein thiol, despite the fact that cellular protein thiols are more abundant than glutathione. Further, these methods usually involve a process of biological sample preparation followed by a separation method, and they are time-consuming. We reported previously a series of thiol-specific fluorogenic benzofurazan sulfides. These nonfluorescent benzofurazan sulfides react rapidly and specifically with a thiol to form a strong fluorescent thiol adduct. The rapid reaction, thiol-specific and fluorogenic nature of the sulfides successfully yielded an application of one of the sulfides for relative quantitation of total thiols in live cells through fluorescence microscopy. In this work, we employed the same compound to develop the first high-throughput method for simultaneous monitoring of protein thiols, nonprotein thiols, and total thiols in cells in a 96-well plate on a fluorescence microplate reader at λex = 430 nm and λem = 520 nm, respectively. The method is rapid and sensitive, and has been validated by an HPLC thiol assay method. The method can detect thiols with cell concentrations as low as 500 cells/well. We also demonstrated that the method can readily monitor changes in cellular thiol levels. Although the method cannot provide an absolute quantification for thiols because fluorescence intensity of different thiol adducts varies, it provides an accurate measurement of relative quantification, relative to the control. The method will be a valuable tool in thiol-related biomedical/pharmaceutical research. PMID:25423115

  9. Hand-Held Reader for Colorimetric Sensor Arrays.

    PubMed

    Askim, Jon R; Suslick, Kenneth S

    2015-08-01

    An inexpensive hand-held device for analysis of colorimetric sensor arrays (CSAs) has been developed. The device makes use of a contact image sensor (CIS), technology commonly used in business card scanners, to rapidly collect low-noise colorimetric data for chemical sensing. The lack of moving parts and insensitivity to vibration allow for lower noise and improved scan rates compared to other digital imaging techniques (e.g., digital cameras, flatbed scanners); signal-to-noise ratios are a factor of 3-10 higher than currently used methods, and scan rates are up to 250 times faster without compromising sensitivity. The device is capable of real-time chemical analysis at scan rates up to 48 Hz. PMID:26177346

  10. Rapid quantification of tilidine, nortilidine, and bisnortilidine in urine by automated online SPE-LC-MS/MS.

    PubMed

    Köhler, Christoph; Grobosch, Thomas; Binscheck, Torsten

    2011-04-01

    The opioid tilidine is a prodrug which is hepatically metabolized to active nortilidine and bisnortilidine. Due to the increasing abuse of tilidine by drug users and the lack of a specific immunoassay, we developed an analytical method for the quantification of tilidine, nortilidine, and bisnortilidine in urine suitable for screening. In a following step, this method was used to establish data on excretion kinetics of the substances in order to evaluate the time window of detection after a single oral dose of tilidine/naloxone and also was applied to authentic urine samples from correctional facilities. Urine samples were mixed with internal standard solution and extracted on a weak cation exchanger at pH 6 using a Symbiosis Pico system. The chromatographic separation was achieved within a 3.5-min run time on a Phenylhexyl column (50 × 2.0 mm, 5 μm) via gradient elution (methanol and 0.2% formic acid) at a flow rate of 0.50 mL/min. The ESI-MS/MS was performed on a QTrap 3,200 in positive multiple reaction monitoring mode using two mass transitions per analyte. Validating the method resulted in a lower limit of quantification of 1.0 μg/L followed by a linear calibration range to 100 μg/L for each analyte (r(2) > 0.99). The analytical method allowed the detection of a single dose of a commercially available tilidine solution up to 7 days after administration. Using this highly sensitive method, 55 of 3,665 urine samples were tested positive. PMID:21140136

  11. Capillary electrophoresis coupled with inductively coupled mass spectrometry as an alternative to cloud point extraction based methods for rapid quantification of silver ions and surface coated silver nanoparticles

    PubMed Central

    Qu, Haiou; Mudalige, Thilak K.; Linder, Sean W.

    2016-01-01

    Speciation and accurate quantification of ionic silver and metallic silver nanoparticles are critical to investigate silver toxicity and to determine the shelf-life of products that contain nano silver under various storage conditions. We developed a rapid method for quantification of silver ions and silver nanoparticles using capillary electrophoresis (CE) interfaced with inductively-coupled plasma mass spectrometry (ICPMS). The addition of 2-mercaptopropionylglycine (tiopronin) to the background electrolyte was used to facilitate the chromatographic separation of ionic silver and maintain the oxidation state of silver. The obtained limits of detection were 0.05 μg kg−1 of silver nanoparticles and 0.03 μg kg−1 of ionic silver. Nanoparticles of varied sizes (10–110 nm) with different surface coating, including citrate acid, lipoic acid, polyvinylpyrrolidone and bovine serum albumin (BSA) were successfully analyzed. Particularly good recoveries (>93%) were obtained for both ionic silver and silver nanoparticle in the presence of excess amount of BSA. The method was further tested with six commercially available dietary supplements which varied in concentration and matrix components. The summed values of silver ions and silver nanoparticles correlated well with the total silver concentration determined by ICPMS after acid digestion. This method can serve as an alternative to cloud point extraction technique when the extraction efficiency for protein coated nanoparticles is low. PMID:26724893

  12. Rapid Quantification of Drug Resistance Gene Expression in Candida albicans by Reverse Transcriptase LightCycler PCR and Fluorescent Probe Hybridization

    PubMed Central

    Frade, Joao P.; Warnock, David W.; Arthington-Skaggs, Beth A.

    2004-01-01

    We developed a rapid, sensitive, and reproducible assay to quantify Candida albicans ACT1, CDR1, CDR2, ERG11, and MDR1 mRNA using a two-step reverse transcription and LightCycler real-time PCR (RT-LightCycler PCR) method with sequence-specific hybridization probes. We compared RT-LightCycler PCR with Northern hybridization for quantitative analysis of gene expression in isolates with various fluconazole susceptibilities. Specificity of each LightCycler PCR was verified by LightCycler melting curve analysis and agarose gel electrophoresis of amplified products. Correlation of quantification results between RT-LightCycler PCR and Northern hybridization yielded correlation coefficients of ≥0.91 for all genes except MDR1 (0.74). In this case, reduced correlation was due to the inability of Northern hybridization to accurately quantify the high MDR1 expression in a susceptible dose-dependent isolate which was shown by RT-LightCycler PCR to overexpress MDR1 >200-fold relative to the other isolates tested. In four isolates, low levels of CDR2 mRNA were detected by RT-LightCycler PCR but were undetectable by Northern hybridization. mRNA quantification by RT-LightCycler PCR correlates with Northern hybridization and offers additional advantages, including increased sensitivity and speed of analysis, along with lower RNA concentration requirements and an increased dynamic range of signal detection. PMID:15131174

  13. Capillary electrophoresis coupled with inductively coupled mass spectrometry as an alternative to cloud point extraction based methods for rapid quantification of silver ions and surface coated silver nanoparticles.

    PubMed

    Qu, Haiou; Mudalige, Thilak K; Linder, Sean W

    2016-01-15

    Speciation and accurate quantification of ionic silver and metallic silver nanoparticles are critical to investigate silver toxicity and to determine the shelf-life of products that contain nano silver under various storage conditions. We developed a rapid method for quantification of silver ions and silver nanoparticles using capillary electrophoresis (CE) interfaced with inductively-coupled plasma mass spectrometry (ICPMS). The addition of 2-mercaptopropionylglycine (tiopronin) to the background electrolyte was used to facilitate the chromatographic separation of ionic silver and maintain the oxidation state of silver. The obtained limits of detection were 0.05 μg kg(-1) of silver nanoparticles and 0.03 μg kg(-1) of ionic silver. Nanoparticles of varied sizes (10-110 nm) with different surface coating, including citrate acid, lipoic acid, polyvinylpyrrolidone and bovine serum albumin (BSA) were successfully analyzed. Particularly good recoveries (>93%) were obtained for both ionic silver and silver nanoparticle in the presence of excess amount of BSA. The method was further tested with six commercially available dietary supplements which varied in concentration and matrix components. The summed values of silver ions and silver nanoparticles correlated well with the total silver concentration determined by ICPMS after acid digestion. This method can serve as an alternative to cloud point extraction technique when the extraction efficiency for protein coated nanoparticles is low. PMID:26724893

  14. Colorimetric and fluorimetric assays to quantitate micromolar concentrations of transition metals.

    PubMed

    McCall, K A; Fierke, C A

    2000-09-10

    Transition metal ions, although maintained at low concentrations, play diverse important roles in many biological processes. Two assays useful for the rapid quantification of a range of first-row transition metal ions have been developed. The colorimetric assay extends the 4-(2-pyridylazo)resorcinol assay of Hunt et al. (J. Biol. Chem. 255, 14793 (1984)) to measure nanomole quantities of Co(2+), Ni(2+), and Cu(2+) as well as Zn(2+). The fluorimetric assay takes advantage of the coordination of a number of metal ions (Mn(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+), Cd(2+)) by Fura-2 and can also be used to measure nanomole quantities of these ions. The assays developed here have the advantage of not requiring the extensive sample preparation necessary for other methodologies, such as atomic absorption spectroscopy and inductively coupled plasma emission spectroscopy (ICPES), while being comparable in accuracy to the detection limits of ICPES for the first-row transition metal ions. To demonstrate the effectiveness of these assays, we determined the affinity of carbonic anhydrase II (CA II), a prototypical zinc enzyme, for Ni(2+) and Cd(2+). These data indicate that CA II binds transition metals with high affinity and is much more selective for Zn(2+) over Ni(2+) or Cd(2+) than most small-molecule chelators or other metalloenzymes. PMID:10964414

  15. Quantification of oxyresveratrol analog trans-2,4,3',5'-tetramethoxystilbene in rat plasma by a rapid HPLC method: application in a pre-clinical pharmacokinetic study.

    PubMed

    Lin, Hai-Shu; Choo, Qiu-Yi; Ho, Paul C

    2010-12-01

    A rapid HPLC method was developed and validated for the quantification of oxyresveratrol analog trans-2,4,3',5'-tetramethoxystilbene (oxyresveratrol tetramethyl ether, OTE) in rat plasma. Chromatographic separation was achieved on an RP-HPLC column, which was protected by a guard column through a 12 min gradient delivery of a mixture of acetonitrile-water at 50°C. The UV absorbance at 325 nm was recorded. The retention time of OTE and trans-stilbene (internal standard) was about 7.7 and 8.4 min, respectively. The calibration curves were linear (R(2) ≥ 0.9986) with a lower limit of quantification of 15 ng/mL. The intra- and inter-day variations, in terms of RSD, were all lower than 9.8% while the intra-day and inter-day bias ranged from -8.3 to +9.2%. The pharmacokinetics of OTE was assessed in rats using 2-hydroxypropyl-β-cyclodextrin as a dosing vehicle. After intravenous administration, OTE possessed a long terminal elimination half-life (t(1/2) (λz) = 481 ± 137 min) and slow clearance (Cl = 29.1 ± 3.7 mL/min/kg). Upon oral administration, OTE was rapidly absorbed. However, it only displayed minimal plasma exposure and its absolute oral bioavailability (F) was as low as 4.5 ± 3.2%. Fortunately, the levels of OTE after single oral administration were sufficient to inhibit human cytochrome P450 1B1. PMID:21077256

  16. Colorimetric Solid-Phase Extractor

    NASA Technical Reports Server (NTRS)

    2003-01-01

    The heart of a colorimetric solid phase extractor (CSPE) test kit quickly measures the concentration of the biocides silver or iodine in astronauts' drinking water to determine whether concentrations are safe. When 10 milliliters (ml) of water is drawn through the disk, the disk will turn color (yellow in this picture for iodine) indicating the presence of the biocides. The device could someday be used to test water safety at reservoirs and water treatment plants on Earth. (photo credit: Microanalytical Instrumentation Center, Iowa State University).

  17. Development of a barcode-style lateral flow immunoassay for the rapid semi-quantification of gliadin in foods.

    PubMed

    Yin, Hsin-Yi; Chu, Pei-Tzu; Tsai, Wen-Che; Wen, Hsiao-Wei

    2016-02-01

    In this work, a barcode-style lateral flow immunoassay is developed using two cut-off values (10 and 50 mg kg(-1) gliadin) to provide a semi-quantification for identifying "gluten-free" and "very low gluten" foods, based on the international Codex Alimentarius Standard. This developed assay exhibits favorable specificity in differentiating wheat from seven commonly used grains, with only a slight cross-reaction with barely. The intra-assay and inter-assay CV values of this assay were 1.5-1.7% and 2.5-4.5%, respectively, revealing high reproducibility. In the analysis of 48 food samples, the results of this assay closely agreed with those obtained using AOAC-approved ELISA or strip kits, as the Cohen's kappa coefficients for both comparisons exceeded 0.8. Thus, this developed assay can be used to quickly estimate the gliadin content in foods in order to protect people with wheat allergy or celiac disease from the accidental ingestion of gliadin. PMID:26304432

  18. Rapid quantification of buprenorphine-glucuronide and norbuprenorphine-glucuronide in human urine by LC-MS-MS.

    PubMed

    Hegstad, S; Khiabani, H Z; Øiestad, E L; Berg, T; Christophersen, A S

    2007-05-01

    A liquid chromatography-tandem mass spectrometry (LC-MS-MS) method was developed and validated for the determination of buprenorphine-glucuronide (BUP-G) and norbuprenorphine-glucuronide (NBUP-G) in human urine. The method included a dilution step followed by filtration through a Mini-Uniprep Filter and direct injection onto the LC column. The analytes were quantified in multiple reactions monitoring mode using one transition ion. Norbuprenorpine-d(3) (NBUP-d(3)) was used as the internal standard. The concentration ranges were 6-161 ng/mL for BUP-G and 12-295 ng/mL for NBUP-G. Recoveries determined after filtration for the analytes were 75%. The between-day precision of the method was in the range of 4.8-11%. The limits of quantification were found to be 4.6 ng/mL for BUP-G and 11.8 ng/mL for NBUP-G. Approximately 1000 samples from law enforcement, prison inmates, probation services, and hospitals were analyzed by the presented method. The ratios of drug glucuronides versus creatinine were calculated for a selection of samples (n = 151), where there was information on treatment with buprenorphine between 16 and 20 mg/day. The majority (86%) of the samples had a ratio of BUP-G/creatinine below 570 microg/g, and 76% of the samples had NBUP-G/creatinine lower than 1060 microg/g. The LC-MS-MS method proved to be robust and specific for the determination of BUP-G and NBUP-G in urine. PMID:17555645

  19. Rapid quantification of miglustat in human plasma and cerebrospinal fluid by liquid chromatography coupled with tandem mass spectrometry.

    PubMed

    Guitton, Jérôme; Coste, Sylvie; Guffon-Fouilhoux, Nathalie; Cohen, Sabine; Manchon, Monique; Guillaumont, Marc

    2009-01-15

    Miglustat (OGT 918) is an iminosugar recently introduced in therapeutic as potential alternative therapy in disorders found in several diseases such as Tay-Sachs, Gaucher or Niemann-Pick diseases. A highly sensitive liquid-chromatography-electrospray tandem mass spectrometry (LC-MS/MS) assay was developed for the quantification of miglustat in human plasma and cerebrospinal fluid (CSF). The sample preparation consists in a simple protein precipitation with a mixture of acetonitrile/methanol (75/25) which yields 100% recovery. The isocratic separation utilizes an Atlantis Hilic (3 microm, 150 mm x 2.1 mm) column, with a mobile phase of acetonitrile/water/ammonium acetate buffer (75/10/15, v/v/v) delivered at 230 microl/min. Selected reaction monitoring (SRM) mode was used with the transitions m/z 220-->158 for the miglustat and m/z 208-->m/z 146 for the miglitol (internal standard). Good linearity was observed in a range from 125 to 2500 ng/ml and from 50 to 1000 ng/ml, for plasma and CSF, respectively. The within-run precision of the assay was less than 6%, and the between-run run precision was less than 6.5%, for six replicates at each of three concentrations and evaluated on three separated days for both plasma and CSF mediums. Assay accuracy was in the range of 98-106.5%. Stability of miglustat was reported under a variety of storage conditions. The miglustat concentrations in two children are presented to demonstrate the clinical interest of this new method. PMID:19095507

  20. A Rapid Protocol of Crude RNA/DNA Extraction for RT-qPCR Detection and Quantification of 'Candidatus Phytoplasma prunorum'

    PubMed Central

    Minguzzi, Stefano; Terlizzi, Federica; Lanzoni, Chiara; Poggi Pollini, Carlo; Ratti, Claudio

    2016-01-01

    Many efforts have been made to develop a rapid and sensitive method for phytoplasma and virus detection. Taking our cue from previous works, different rapid sample preparation methods have been tested and applied to Candidatus Phytoplasma prunorum (‘Ca. P. prunorum’) detection by RT-qPCR. A duplex RT-qPCR has been optimized using the crude sap as a template to simultaneously amplify a fragment of 16S rRNA of the pathogen and 18S rRNA of the host plant. The specific plant 18S rRNA internal control allows comparison and relative quantification of samples. A comparison between DNA and RNA contribution to qPCR detection is provided, showing higher contribution of the latter. The method presented here has been validated on more than a hundred samples of apricot, plum and peach trees. Since 2013, this method has been successfully applied to monitor ‘Ca. P. prunorum’ infections in field and nursery. A triplex RT-qPCR assay has also been optimized to simultaneously detect ‘Ca. P. prunorum’ and Plum pox virus (PPV) in Prunus. PMID:26742106

  1. Development of a Rapid Cartilage Damage Quantification Method for the Lateral Tibiofemoral Compartment Using Magnetic Resonance Images: Data from the Osteoarthritis Initiative

    PubMed Central

    Zhang, Ming; Driban, Jeffrey B.; Price, Lori Lyn; Lo, Grace H.; Miller, Eric; McAlindon, Timothy E.

    2015-01-01

    The purpose of this study was to expand and validate the cartilage damage index (CDI) to detect cartilage damage in the lateral tibiofemoral compartment. We used an iterative 3-step process to develop and validate the lateral CDI: development (100 knees), testing (80 knees), and validation (100 knees). The validation set included 100 knees from the Osteoarthritis Initiative that was enriched to include all grades of lateral joint space narrowing (JSN, 0–3). Measurement of the CDI was rapid at 7.4 (s.d. 0.73) minutes per knee pair (baseline and follow-up of one knee). The intratester reliability is good (intraclass correlation coefficient (3, 1 model) = 0.86 to 0.98). At baseline, knees with greater KL grade and lateral JSN had a lower mean CDI (i.e., greater cartilage damage). Baseline lateral CDI is associated with both lateral JSW (r = 0.81 to 0.85, p < 0.01) and HKA (r = −0.30 to −0.33, p < 0.05). The SRM is good (lateral femur SRM = −0.76; lateral tibia SRM = −0.73; lateral tibiofemoral total SRM = −0.87). The lateral tibiofemoral CDI quantification allows for rapid evaluation and is reliable and responsive, with good construct validity. It may be an efficient method to measure lateral tibiofemoral articular cartilage in large clinical and epidemiologic studies. PMID:26713316

  2. A Rapid Protocol of Crude RNA/DNA Extraction for RT-qPCR Detection and Quantification of 'Candidatus Phytoplasma prunorum'.

    PubMed

    Minguzzi, Stefano; Terlizzi, Federica; Lanzoni, Chiara; Poggi Pollini, Carlo; Ratti, Claudio

    2016-01-01

    Many efforts have been made to develop a rapid and sensitive method for phytoplasma and virus detection. Taking our cue from previous works, different rapid sample preparation methods have been tested and applied to Candidatus Phytoplasma prunorum ('Ca. P. prunorum') detection by RT-qPCR. A duplex RT-qPCR has been optimized using the crude sap as a template to simultaneously amplify a fragment of 16S rRNA of the pathogen and 18S rRNA of the host plant. The specific plant 18S rRNA internal control allows comparison and relative quantification of samples. A comparison between DNA and RNA contribution to qPCR detection is provided, showing higher contribution of the latter. The method presented here has been validated on more than a hundred samples of apricot, plum and peach trees. Since 2013, this method has been successfully applied to monitor 'Ca. P. prunorum' infections in field and nursery. A triplex RT-qPCR assay has also been optimized to simultaneously detect 'Ca. P. prunorum' and Plum pox virus (PPV) in Prunus. PMID:26742106

  3. Adobe photoshop quantification (PSQ) rather than point-counting: A rapid and precise method for quantifying rock textural data and porosities

    NASA Astrophysics Data System (ADS)

    Zhang, Xuefeng; Liu, Bo; Wang, Jieqiong; Zhang, Zhe; Shi, Kaibo; Wu, Shuanglin

    2014-08-01

    Commonly used petrological quantification methods are visual estimation, counting, and image analyses. However, in this article, an Adobe Photoshop-based analyzing method (PSQ) is recommended for quantifying the rock textural data and porosities. Adobe Photoshop system provides versatile abilities in selecting an area of interest and the pixel number of a selection could be read and used to calculate its area percentage. Therefore, Adobe Photoshop could be used to rapidly quantify textural components, such as content of grains, cements, and porosities including total porosities and different genetic type porosities. This method was named as Adobe Photoshop Quantification (PSQ). The workflow of the PSQ method was introduced with the oolitic dolomite samples from the Triassic Feixianguan Formation, Northeastern Sichuan Basin, China, for example. And the method was tested by comparing with the Folk's and Shvetsov's "standard" diagrams. In both cases, there is a close agreement between the "standard" percentages and those determined by the PSQ method with really small counting errors and operator errors, small standard deviations and high confidence levels. The porosities quantified by PSQ were evaluated against those determined by the whole rock helium gas expansion method to test the specimen errors. Results have shown that the porosities quantified by the PSQ are well correlated to the porosities determined by the conventional helium gas expansion method. Generally small discrepancies (mostly ranging from -3% to 3%) are caused by microporosities which would cause systematic underestimation of 2% and/or by macroporosities causing underestimation or overestimation in different cases. Adobe Photoshop could be used to quantify rock textural components and porosities. This method has been tested to be precise and accurate. It is time saving compared with usual methods.

  4. Rapid, absolute, and simultaneous quantification of specific pathogenic strain and total bacterial cells using an ultrasensitive dual-color flow cytometer.

    PubMed

    Yang, Lingling; Wu, Lina; Zhu, Shaobin; Long, Yao; Hang, Wei; Yan, Xiaomei

    2010-02-01

    This paper describes a rapid and sensitive strategy for the absolute and simultaneous quantification of specific pathogenic strain and total bacterial cells in a mixture. A laboratory-built compact, high-sensitivity, dual channel flow cytometer (HSDCFCM) was modified to enable dual fluorescence detection. A bacterial cell mixture comprising heat-killed pathogenic Escherichia coli E. coli O157:H7 and harmless E. coli DH5alpha was used as a model system. Pathogenic E. coli O157:H7 cells were selectively labeled by red fluorescent probe via antibody-antigen interaction, and all bacterial cells were stained with membrane-permeable nucleic acid dye that fluoresces green. When each individual bacterium passes through the interrogating laser beam, E. coli O157:H7 emits both red and green fluorescence, while E. coli DH5alpha exhibits only green fluorescence. Because the fluorescence burst generated from each individual bacterial cell was easily distinguished from the background, accurate enumeration and consequently absolute quantification were achieved for both pathogenic and total bacterial cells. By using this strategy, accurate counting of bacteria at a density above 1.0 x 10(5) cells/mL can be accomplished with 1 min of data acquisition time after fluorescent staining. Excellent correlation between the concentrations measured by the HSDCFCM and the conventional plate-counting method were obtained for pure-cultured E. coli O157:H7 (R(2) = 0.9993) and E. coli DH5alpha (R(2) = 0.9998). Bacterial cell mixtures with varying proportions of E. coli O157:H7 and E. coli DH5alpha were measured with good ratio correspondence. We applied the established approach to detecting artificially contaminated drinking water samples; E. coli O157:H7 of 1.0 x 10(2) cells/mL were accurately quantified upon sample enrichment. It is believed that the proposed method will find wide applications in many fields demanding bacterial identification and quantification. PMID:20039721

  5. Rapid detection and quantification of tumor marker carbohydrate antigen 72-4 (CA72-4) using a superparamagnetic immunochromatographic strip.

    PubMed

    Chen, Yanrong; Wang, Kan; Liu, Zongrui; Sun, Rongjin; Cui, Daxiang; He, Jinghua

    2016-03-01

    Rapid and quantitative detection of biomarkers with high sensitivity and specificity has become a common practice in the clinical diagnosis and treatment of cancer. Herein, a quantitative lateral flow immunoassay (LFIA) based on superparamagnetic nanoparticles (SPMNPs) was developed for detection of carbohydrate antigen 72-4 (CA72-4) in human serum. This direct and rapid method did not involve any sample preparation and was accomplished using a test strip in which a sandwich reaction was performed. Probes on the conjugate pad were prepared by coupling monoclonal antibody CC49 specific to CA72-4 onto SPMNPs. Coupled monoclonal antibody B72.3 and goat anti-mouse IgG were loaded onto a nitrocellulose (NC) membrane, serving as the test and control lines, respectively. Initially, results were evaluated by macroscopic observation. Afterwards, the magnetic signal strength of the reaction area was quantified using a magnetic assay reader (MAR). Several parameters that may influence the detection sensitivity were studied and optimized. Under optimal conditions, the proposed method was capable of detecting as low as 0.38 IU/mL of CA72-4 in 20 min and had a wide detection linearity range (0-100 IU/mL). We evaluated 100 clinical samples (70 positive and 30 negative) to assess the validity of these test strips, which exhibited high sensitivity (99 %) and specificity (97 %). The results indicated a high rate of accuracy (98.4-102 %) and a low relative standard deviation according to the average recovery test. In conclusion, the test strips based on SPMNP probes are a rapid, sensitive, and quantitative method for the detection of CA72-4 and possess great potential in point-of-care testing (POCT). Graphical Abstract Schematic illustration of the strategy for CA72-4 detection and quantification using a superparamagnetic immunochromatographic strip. PMID:26825343

  6. Application of a rapid scanning plasma emission detector and gas chromatography for multi-element quantification of halogenated hydrocarbons.

    PubMed

    Zerezghi, M; Mulligan, K J; Caruso, J A

    1984-08-01

    A microwave induced plasma emission detector is used as an element-selective detector for gas chromatography. The spectrometer, which is fitted with a rapid scanning galvanometer mirror, is used to scan a pre-selected spectral window to provide information in the multi-element mode. This information is used to determine the per mole response of some elements as a function of molecular structure. Despite the low microwave powers employed, the response per mole appears to be independent of the molecular structure. Detection limits and linear dynamic ranges are determined by narrowing the spectral coverage to increase the sensitivity. Calibration curves are linear over several orders of magnitude and detection limits are at the pg/sec levels. PMID:6480798

  7. Photoacoustic and Colorimetric Visualization of Latent Fingerprints.

    PubMed

    Song, Kai; Huang, Peng; Yi, Chenglin; Ning, Bo; Hu, Song; Nie, Liming; Chen, Xiaoyuan; Nie, Zhihong

    2015-12-22

    There is a high demand on a simple, rapid, accurate, user-friendly, cost-effective, and nondestructive universal method for latent fingerprint (LFP) detection. Herein, we describe a combination imaging strategy for LFP visualization with high resolution using poly(styrene-alt-maleic anhydride)-b-polystyrene (PSMA-b-PS) functionalized gold nanoparticles (GNPs). This general approach integrates the merits of both colorimetric imaging and photoacoustic imaging. In comparison with the previous methods, our strategy is single-step and does not require the signal amplification by silver staining. The PSMA-b-PS functionalized GNPs have good stability, tunable color, and high affinity for universal secretions (proteins/polypeptides/amino acids), which makes our approach general and flexible for visualizing LFPs on different substrates (presumably with different colors) and from different people. Moreover, the unique optical property of GNPs enables the photoacoustic imaging of GNPs-deposited LFPs with high resolution. This allows observation of level 3 hyperfine features of LFPs such as the pores and ridge contours by photoacoustic imaging. This technique can potentially be used to identify chemicals within LFP residues. We believe that this dual-modality imaging of LFPs will find widespread use in forensic investigations and medical diagnostics. PMID:26528550

  8. Development of a colony lift immunoassay to facilitate rapid detection and quantification of Escherichia coli O157:H7 from agar plates and filter monitor membranes.

    PubMed

    Ingram, D T; Lamichhane, C M; Rollins, D M; Carr, L E; Mallinson, E T; Joseph, S W

    1998-07-01

    E. coli O157:H7 is a food-borne adulterant that can cause hemorrhagic ulcerative colitis and hemolytic uremic syndrome. Faced with an increasing risk of foods contaminated with E. coli O157:H7, food safety officials are seeking improved methods to detect and isolate E. coli O157:H7 in hazard analysis and critical control point systems in meat- and poultry-processing plants. A colony lift immunoassay was developed to facilitate the positive identification and quantification of E. coli O157:H7 by incorporating a simple colony lift enzyme-linked immunosorbent assay with filter monitors and traditional culture methods. Polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, Mass.) were prewet with methanol and were used to make replicates of every bacterial colony on agar plates or filter monitor membranes that were then reincubated for 15 to 18 h at 36 +/- 1 degree C, during which the colonies not only remained viable but were reestablished. The membranes were dried, blocked with blocking buffer (Kirkegaard and Perry Laboratories [KPL], Gaithersburg, Md.), and exposed for 7 min to an affinity-purified horseradish peroxidase-labeled goat anti-E. coli O157 antibody (KPL). The membranes were washed, exposed to a 3,3',5,5'-tetramethylbenzidine membrane substrate (TMB; KPL) or aminoethyl carbazole (AEC; Sigma Chemical Co., St. Louis, Mo.), rinsed in deionized water, and air dried. Colonies of E. coli O157:H7 were identified by either a blue (via TMB) or a red (via AEC) color reaction. The colored spots on the PVDF lift membrane were then matched to their respective parent colonies on the agar plates or filter monitor membranes. The colony lift immunoassay was tested with a wide range of genera in the family Enterobacteriaceae as well as different serotypes within the E. coli genus. The colony lift immunoassay provided a simple, rapid, and accurate method for confirming the presence of E. coli O157:H7 colonies isolated on filter monitors or spread plates by

  9. A study on the use of near-infrared spectroscopy for the rapid quantification of major compounds in Tanreqing injection

    NASA Astrophysics Data System (ADS)

    Li, Wenlong; Cheng, Zhiwei; Wang, Yuefei; Qu, Haibin

    2013-01-01

    In this paper we describe the strategy used in the development and validation of a near infrared spectroscopy method for the rapid determination of baicalin, chlorogenic acid, ursodeoxycholic acid (UDCA), chenodeoxycholic acid (CDCA), and the total solid contents (TSCs) in the Tanreqing injection. To increase the representativeness of calibration sample set, a concentrating-diluting method was adopted to artificially prepare samples. Partial least square regression (PLSR) was used to establish calibration models, with which the five quality indicators can be determined with satisfied accuracy and repeatability. In addition, the slope/bias (S/B) method was used for the models transfer between two different types of NIR instruments from the same manufacturer, which is contributing to enlarge the application range of the established models. With the presented method, a great deal of time, effort and money can be saved when large amounts of Tanreqing injection samples need to be analyzed in a relatively short period of time, which is of great significance to the traditional Chinese medicine (TCM) industries.

  10. Two novel methods for rapid detection and quantification of DNMT3A R882 mutations in acute myeloid leukemia.

    PubMed

    Mancini, Melissa; Hasan, Syed Khizer; Ottone, Tiziana; Lavorgna, Serena; Ciardi, Claudia; Angelini, Daniela F; Agostini, Francesca; Venditti, Adriano; Lo-Coco, Francesco

    2015-03-01

    DNMT3A mutations represent one of the most frequent gene alterations detectable in acute myeloid leukemia with normal karyotype. Although various recurrent somatic mutations of DNMT3A have been described, the most common mutation is located at amino acid R882 in the methyltransferase domain of the gene. DNMT3A mutations have been reported to be stable during disease progression and are associated with unfavorable outcome in acute myeloid leukemia patients with normal karyotype. Because of their prognostic significance and high stability during disease evolution, DNMT3A mutations might represent highly informative biomarkers for minimal residual disease monitoring. We describe a new rapid diagnostic RT-PCR assay based on TauI restriction enzyme reaction to identify DNMT3A R882 mutations at diagnosis. In addition, we developed a sensitive and specific test based on peptide nucleic acid real-time PCR technology to monitor DNMT3A R882H mutation. We identified 24 DNMT3A R882H mutated patients out of 134 acute myeloid leukemia screened samples and we analyzed in these patients the kinetics of minimal residual disease after induction and consolidation therapy. This assay may be useful to better assess response to therapy in patients with acute myeloid leukemia bearing the DNMT3A R882H mutation. PMID:25554589

  11. Surfactant-based ionic liquids for extraction of phenolic compounds combined with rapid quantification using capillary electrophoresis.

    PubMed

    Huang, Fangzhi; Berton, Paula; Lu, Chengfei; Siraj, Noureen; Wang, Chun; Magut, Paul K S; Warner, Isiah M

    2014-09-01

    A rapid liquid phase extraction employing a novel hydrophobic surfactant-based room temperature ionic liquid (RTIL), tetrabutylphosphonium dioctyl sulfosuccinate ([4C4 P][AOT]), coupled with capillary electrophoretic-UV (CE-UV) detection is developed for removal and determination of phenolic compounds. The long-carbon-chain RTIL used is sparingly soluble in most solvents and can be used to replace volatile organic solvents. This fact, in combination with functional-surfactant-anions, is proposed to reduce the interfacial energy of the two immiscible liquid phases, resulting in highly efficient extraction of analytes. Several parameters that influence the extraction efficiencies, such as extraction time, RTIL type, pH value, and ionic strength of aqueous solutions, were investigated. It was found that, under acidic conditions, most of the investigated phenols were extracted from aqueous solution into the RTIL phase within 12 min. Good linearity was observed over the concentration range of 0.1-80.0 μg/mL for all phenols investigated. The precision of this method, expressed as RSD, was determined to be within 3.4-5.3% range. The LODs (S/N = 3) of the method were in the range of 0.047-0.257 μg/mL. The proposed methodology was successfully applied to determination of phenols in real water samples. PMID:24798689

  12. QuShape: Rapid, accurate, and best-practices quantification of nucleic acid probing information, resolved by capillary electrophoresis

    PubMed Central

    Karabiber, Fethullah; McGinnis, Jennifer L.; Favorov, Oleg V.; Weeks, Kevin M.

    2013-01-01

    Chemical probing of RNA and DNA structure is a widely used and highly informative approach for examining nucleic acid structure and for evaluating interactions with protein and small-molecule ligands. Use of capillary electrophoresis to analyze chemical probing experiments yields hundreds of nucleotides of information per experiment and can be performed on automated instruments. Extraction of the information from capillary electrophoresis electropherograms is a computationally intensive multistep analytical process, and no current software provides rapid, automated, and accurate data analysis. To overcome this bottleneck, we developed a platform-independent, user-friendly software package, QuShape, that yields quantitatively accurate nucleotide reactivity information with minimal user supervision. QuShape incorporates newly developed algorithms for signal decay correction, alignment of time-varying signals within and across capillaries and relative to the RNA nucleotide sequence, and signal scaling across channels or experiments. An analysis-by-reference option enables multiple, related experiments to be fully analyzed in minutes. We illustrate the usefulness and robustness of QuShape by analysis of RNA SHAPE (selective 2′-hydroxyl acylation analyzed by primer extension) experiments. PMID:23188808

  13. Zinc Biosorption by Seaweed Illustrated by the Zincon Colorimetric Method and the Langmuir Isotherm

    ERIC Educational Resources Information Center

    Areco, Maria Mar; dos Santos Afonso, Maria; Valdman, Erika

    2007-01-01

    An experiment is conducted to promote biotechnology knowledge that is an emerging technology on cleaning treatment, showing the potential of seaweed to remove heavy-metal ions from solution. The rapid and accurate determination of zinc in aqueous solution by the zincon colorimetric method gives an interesting and simple experiment for any…

  14. Identification of Escherichia coli O157 by Using a Novel Colorimetric Detection Method with DNA Microarrays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Shiga toxin-producing Escherichia coli O157:H7 is a leading cause of foodborne illness worldwide. To evaluate better methods to rapidly detect and genotype E. coli O157 strains, the present study evaluated the use of ampliPHOX, a novel colorimetric detection method based on photopolymerization, for...

  15. Novel, Rapid Identification, and Quantification of Adulterants in Extra Virgin Olive Oil Using Near-Infrared Spectroscopy and Chemometrics.

    PubMed

    Azizian, Hormoz; Mossoba, Magdi M; Fardin-Kia, Ali Reza; Delmonte, Pierluigi; Karunathilaka, Sanjeewa R; Kramer, John K G

    2015-07-01

    A new, rapid Fourier transform near infrared (FT-NIR) spectroscopic procedure is described to screen for the authenticity of extra virgin olive oils (EVOO) and to determine the kind and amount of an adulterant in EVOO. To screen EVOO, a partial least squares (PLS1) calibration model was developed to estimate a newly created FT-NIR index based mainly on the relative intensities of two unique carbonyl overtone absorptions in the FT-NIR spectra of EVOO and other mixtures attributed to volatile (5280 cm(-1)) and non-volatile (5180 cm(-1)) components. Spectra were also used to predict the fatty acid (FA) composition of EVOO or samples spiked with an adulterant using previously developed PLS1 calibration models. Some adulterated mixtures could be identified provided the FA profile was sufficiently different from those of EVOO. To identify the type and determine the quantity of an adulterant, gravimetric mixtures were prepared by spiking EVOO with different concentrations of each adulterant. Based on FT-NIR spectra, four PLS1 calibration models were developed for four specific groups of adulterants, each with a characteristic FA composition. Using these different PLS1 calibration models for prediction, plots of predicted vs. gravimetric concentrations of an adulterant in EVOO yielded linear regression functions with four unique sets of slopes, one for each group of adulterants. Four corresponding slope rules were defined that allowed for the determination of the nature and concentration of an adulterant in EVOO products by applying these four calibration models. The standard addition technique was used for confirmation. PMID:26050093

  16. Rapid diagnosis and quantification of Francisella tularensis in organs of naturally infected common squirrel monkeys (Saimiri sciureus).

    PubMed

    Abril, Carlos; Nimmervoll, Helena; Pilo, Paola; Brodard, Isabelle; Korczak, Bozena; Markus, Seiler; Miserez, Raymond; Frey, Joachim

    2008-02-01

    Francisella tularensis, a small Gram-negative facultative intracellular bacterium, is the causative agent of tularaemia, a severe zoonotic disease transmitted to humans mostly by vectors such as ticks, flies and mosquitoes. The disease is endemic in many parts of the northern hemisphere. Among animals, the most affected species belong to rodents and lagomorphs, in particular hares. However, in the recent years, many cases of tularaemia among small monkeys in zoos were reported. We have developed a real-time PCR that allows to quantify F. tularensis in tissue samples. Using this method, we identified the spleen and the kidney as the most heavily infected organ containing up to 400 F. tularensis bacteria per simian host cell in two common squirrel monkeys (Saimiri sciureus) from a zoo that died of tularaemia. In other organs such as the brain, F. tularensis was detected at much lower titres. The strain that caused the infection was identified as F. tularensis subsp. holarctica biovar I, which is susceptible to erythromycin. The high number of F. tularensis present in soft organs such as spleen, liver and kidney represents a high risk for persons handling such carcasses and explains the transmission of the disease to a pathologist during post-mortem analysis. Herein, we show that real-time PCR allows a reliable and rapid diagnosis of F. tularensis directly from tissue samples of infected animals, which is crucial in order to attempt accurate prophylactic measures, especially in cases where humans or other animals have been exposed to this highly contagious pathogen. PMID:17875369

  17. Sensitive and rapid analytical method for the quantification of glucosamine in human plasma by ultra high performance liquid chromatography with tandem mass spectrometry.

    PubMed

    Yang, Wen; Zheng, Xiaohong; Simpemba, Ernest; Ma, Pengcheng; Ding, Li

    2015-06-01

    A highly sensitive and rapid ultra high performance liquid chromatography with tandem mass spectrometry method has been developed and validated for the determination of glucosamine in human plasma using miglitol as the internal standard. Special attention was paid to achieve the high throughput and sensitivity of the established method, and the absence of a matrix effect on the analytes. The sample preparation procedure involved a simple deproteinization step. The chromatographic separation was achieved on a Waters ACQUITY HSS Cyano column using a mixture of acetonitrile/2 mM ammonium acetate solution containing 0.03% formic acid (80:20, v/v) as the mobile phase with a very short run time of 1.5 min. This method was validated over the concentration range of 10-3000 ng/mL for glucosamine. The intra- and inter-batch precision was <13.9% for the low, medium, and high quality control samples. The established method is highly sensitive with a lower limit of quantification of 10 ng/mL, low enough to determine the circadian rhythm on endogenous glucosamine level in human plasma, which has not been reported in detail until now. The method was successfully applied to characterize the pharmacokinetic profile of glucosamine in healthy volunteers following a single oral administration of 750 or 1500 mg glucosamine hydrochloride. PMID:25802209

  18. Combination of crossflow ultrafiltration, monolithic affinity filtration, and quantitative reverse transcriptase PCR for rapid concentration and quantification of model viruses in water.

    PubMed

    Pei, Lu; Rieger, Martin; Lengger, Sandra; Ott, Sonja; Zawadsky, Claudia; Hartmann, Nils Marten; Selinka, Hans-Christoph; Tiehm, Andreas; Niessner, Reinhard; Seidel, Michael

    2012-09-18

    We present a rapid and effective adsorption-elution method based on monolithic affinity filtration (MAF) for the concentration and purification of waterborne viruses. The MAF column consists of a hydrolyzed macroporous epoxy-based polymer. High recoveries were achieved by columns for the bacterial virus (bacteriophage) MS2 110 (±19)%, as model organism, as well as for human adenoviruses 42.4 (±3.4)% and murine noroviruses 42.6 (±1.9)%. This new concentration and purification method was combined with crossflow ultrafiltration (CUF). Because of the adsorption of the examined viruses to the macroporous surface of the MAF column at pH 3, concentrated matrix components by CUF can be removed. Bacteriophages MS2 were spiked in tap water and concentrated with the new CUF-MAF concentration method by a volumetric factor of 10(4) within 33 min. Furthermore, the detection limit for quantification of bacteriophage MS2 by quantitative reverse transcriptase PCR (qRT-PCR) could be improved from 79.47 to 0.0056 GU mL(-1) by a factor of 1.4 × 10(4). In a first study, we have shown that this method could also be applied for river water containing naturally MS2 and MS2-like phages. PMID:22917471

  19. Development of a Taqman real-time PCR assay for rapid detection and quantification of Vibrio tapetis in extrapallial fluids of clams

    PubMed Central

    Richard, Gaëlle G.; Le Bris, Cédric; Paillard, Christine

    2015-01-01

    The Gram-negative bacterium Vibrio tapetis is known as the causative agent of Brown Ring Disease (BRD) in the Manila clam Venerupis (=Ruditapes) philippinarum. This bivalve is the second most important species produced in aquaculture and has a high commercial value. In spite of the development of several molecular methods, no survey has been yet achieved to rapidly quantify the bacterium in the clam. In this study, we developed a Taqman real-time PCR assay targeting virB4 gene for accurate and quantitative identification of V. tapetis strains pathogenic to clams. Sensitivity and reproducibility of the method were assessed using either filtered sea water or extrapallial fluids of clam injected with the CECT4600T V. tapetis strain. Quantification curves of V. tapetis strain seeded in filtered seawater (FSW) or extrapallial fluids (EF) samples were equivalent showing reliable qPCR efficacies. With this protocol, we were able to specifically detect V. tapetis strains down to 1.125 101 bacteria per mL of EF or FSW, taking into account the dilution factor used for appropriate template DNA preparation. This qPCR assay allowed us to monitor V. tapetis load both experimentally or naturally infected Manila clams. This technique will be particularly useful for monitoring the kinetics of massive infections by V. tapetis and for designing appropriate control measures for aquaculture purposes. PMID:26713238

  20. Rapid and selective quantification of L-theanine in ready-to-drink teas from Chinese market using SPE and UPLC-UV.

    PubMed

    Chen, Guoqiang; Wang, Yun; Song, Weiqi; Zhao, Bo; Dou, Yuling

    2012-11-15

    An ultra performance liquid chromatography (UPLC) method combined with solid phase extraction (SPE) sample pre-treatment was developed and validated for the rapid quantification of L-theanine in ready-to-drink (RTD) teas. UPLC analysis of twenty-seven RTD teas from the Chinese market revealed that the L-theanine levels in various types of RTD teas were significantly different. RTD green teas were found to contain highest mean L-theanine level (37.85±20.54 mg/L), followed by jasmine teas (36.60±12.08 mg/L), Tieguanying teas (18.54±3.46 mg/L) black teas (16.89±6.56), Pu-erh teas (11.31±0.90 mg/L) and oolong teas (3.85±2.27 mg/L). The ratio of total polyphenols content to L-theanine content could be used as a featured parameter for differentiating RTD teas. L-theanine in RTD teas could be a reliable quality parameter that is complementary to total polyphenols. PMID:22868106

  1. A rapid and sensitive LC-MS/MS method for quantification of 3,29-dibenzoyl rarounitriol in rat plasma: application to a pharmacokinetic study.

    PubMed

    Zhao, Chengliang; Zhang, Nan; Chen, Bin; Song, Youxin; Zhu, Naiqiang; Zhao, Long; Liu, Changjian; Meng, Xiangwei

    2015-08-01

    A rapid, sensitive and high-throughput liquid chromatography-tandem mass spectrometry was established and validated to assay the concentrations of 3,29-dibenzoyl rarounitriol in rat plasma. Plasma samples were processed by liquid-liquid extraction with ethyl acetate and separated on a Hypersil Gold C18 column (50 × 4.6 mm, 3 µm) at an isocratic flow rate of 0.5 mL/min using methanol-10 mm ammonium acetate-formic acid (90:10:0.1, v/v/v) as mobile phase. The total run time was 5 min for each sample. MS/MS detection was accomplished in selected reaction monitoring mode with positive electrospray ionization. The calibration curve was linear over the concentration range of 0.125-50 ng/mL with lower limit of quantification of 0.125 ng/mL. The intra- and inter-day precisions were <10.1% in terms of coefficient of variation, and the accuracy was within ±11.7% in terms of relative error. The developed method was successfully applied to a pharmacokinetic study of 3,29-dibenzoyl rarounitriol following intragastric administration of 3.65 mg/kg to Wistar rats. PMID:25611485

  2. Simple and rapid determination of phthalates using microextraction by packed sorbent and gas chromatography with mass spectrometry quantification in cold drink and cosmetic samples.

    PubMed

    Kaur, Ramandeep; Heena; Kaur, Ripneel; Rani, Susheela; Malik, Ashok Kumar

    2016-03-01

    A simple and rapid method using microextraction by packed sorbent coupled with gas chromatography and mass spectrometry has been developed for the analysis of five phthalates, namely, diethyl phthalate, benzyl-n-butyl phthalate, dicyclohexyl phthalate, di-n-butyl phthalate, and di-n-propyl phthalate, in cold drink and cosmetic samples. The various parameters that influence the microextraction by packed sorbent performance such as extraction cycle (extract-discard), type and amount of solvent, washing solvent, and pH have been studied. The optimal conditions of microextraction using C18 as the packed sorbent were 15 extraction cycles with water as washing solvent and 3 × 10 μL of ethyl acetate as the eluting solvent. Chromatographic separation was also optimized for injection temperature, flow rate, ion source, interface temperature, column temperature gradient and mass spectrometry was evaluated using the scan and selected ion monitoring data acquisition mode. Satisfactory results were obtained in terms of linearity with R(2) >0.9992 within the established concentration range. The limit of detection was 0.003-0.015 ng/mL, and the limit of quantification was 0.009-0.049 ng/mL. The recoveries were in the range of 92.35-98.90% for cold drink, 88.23-169.20% for perfume, and 88.90-184.40% for cream. Analysis by microextraction by packed sorbent promises to be a rapid method for the determination of these phthalates in cold drink and cosmetic samples, reducing the amount of sample, solvent, time and cost. PMID:26683135

  3. Highly Sensitive Colorimetric Detection of Ochratoxin A by a Label-Free Aptamer and Gold Nanoparticles

    PubMed Central

    Luan, Yunxia; Chen, Jiayi; Li, Cheng; Xie, Gang; Fu, Hailong; Ma, Zhihong; Lu, Anxiang

    2015-01-01

    A label-free aptamer-based assay for the highly sensitive and specific detection of Ochratoxin A (OTA) was developed using a cationic polymer and gold nanoparticles (AuNPs). The OTA aptamer was used as a recognition element for the colorimetric detection of OTA based on the aggregation of AuNPs by the cationic polymer. By spectroscopic quantitative analysis, the colorimetric assay could detect OTA down to 0.009 ng/mL with high selectivity in the presence of other interfering toxins. This study offers a new alternative in visual detection methods that is rapid and sensitive for OTA detection. PMID:26690477

  4. Rapid analysis and quantification of fluorescent brighteners in wheat flour by Tri-step infrared spectroscopy and computer vision technology

    NASA Astrophysics Data System (ADS)

    Guo, Xiao-Xi; Hu, Wei; Liu, Yuan; Gu, Dong-Chen; Sun, Su-Qin; Xu, Chang-Hua; Wang, Xi-Chang

    2015-11-01

    Fluorescent brightener, industrial whitening agent, has been illegally used to whitening wheat flour. In this article, computer vision technology (E-eyes) and colorimetry were employed to investigate color difference among different concentrations of fluorescent brightener in wheat flour using DMS as an example. Tri-step infrared spectroscopy (Fourier transform-infrared spectroscopy coupled with second derivative infrared spectroscopy (SD-IR) and two dimensional correlation infrared spectroscopy (2DCOS-IR)) was used to identify and quantitate DMS in wheat flour. According to color analysis, the whitening effect was significant when added with less than 30 mg/g DMS but when more than 100 mg/g, the flour began greenish. Thus it was speculated that the concentration of DMS should be below 100 mg/g in real flour adulterant with DMS. With the increase of the concentration, the spectral similarity of wheat flour with DMS to DMS standard was increasing. SD-IR peaks at 1153 cm-1, 1141 cm-1, 1112 cm-1, 1085 cm-1 and 1025 cm-1 attributed to DMS were regularly enhanced. Furthermore, it could be differentiated by 2DOS-IR between DMS standard and wheat flour added with DMS low to 0.05 mg/g and the bands in the range of 1000-1500 cm-1 could be an exclusive range to identify whether wheat flour contained DMS. Finally, a quantitative prediction model based on IR spectra was established successfully by Partial least squares (PLS) with a concentration range from 1 mg/g to 100 mg/g. The calibration set gave a determination coefficient of 0.9884 with a standard error (RMSEC) of 5.56 and the validation set presented a determination coefficient of 0.9881 with a standard error of 5.73. It was demonstrated that computer vision technology and colorimetry were effective to estimate the content of DMS in wheat flour and the Tri-step infrared macro-fingerprinting combined with PLS was applicable for rapid and nondestructive fluorescent brightener identification and quantitation.

  5. A rapid and highly sensitive UPLC-MS-MS method for the quantification of zolpidem tartrate in human EDTA plasma and its application to pharmacokinetic study.

    PubMed

    Reddy, Dendhi Chandrapal; Bapuji, Akula Tukaram; Rao, Vepakomma Suryanarayana; Himabindu, Vurimindi; Ravinder, Sreedasyam

    2012-07-01

    A rapid and high sensitive liquid chromatography-tandem mass spectrometry (LC-MS-MS) method was developed and validated for the quantification of zolpidem in human EDTA plasma using ondansetron (IS) as an internal standard. The analyte and IS were extracted from human plasma using ethyl acetate and separated on a C18 column (Inertsil-ODS, 5 µm, 4.6 × 50 mm) interfaced with a triple quadrupole tandem mass spectrometer. The mobile phase, which consisted of a mixture of methanol and 20 mM ammonium formate (pH 5.00 ± 0.05; 75:25 v/v), was injected at a flow rate of 0.40 mL/min. The retention times of zolpidem and IS were approximately 1.76 and 1.22. The LC run time was 3 min. The electrospray ionization source was operated in positive ion mode. Multiple reaction monitoring used the [M + H](+) ions m/z 308.13 → 235.21 for zolpidem and m/z 294.02 → 170.09 for the ondansetron, respectively. Five freeze-thaw cycles was established at -20 and -70°C.The linearity of the response/concentration curve was established in human EDTA plasma over the concentration range 0.10-149.83 ng/mL. The lower detection limit [(signal-to-noise (S/N) > 3] was 0.04 ng/mL and the lower limit of quantification (S/N > 10) was 0.10 ng/mL. This LC-MS-MS method was validated with intra-batch and inter-batch precision of 0.52-8.66.The intra-batch and inter-batch accuracy was 96.66-106.11. Recovery of zolpidem in human plasma was 87.00% and IS recovery was 81.60%. The primary pharmacokinetic parameters were T(max) (h) = (1.25 ± 0.725), C(max) (ng/mL) (127.80 ± 34.081), AUC(0→t), = (665.37 ± 320.982) and AUC(0→∞), 686.03 ± 342.952, respectively. PMID:22689921

  6. Gold nanoparticles based colorimetric nanodiagnostics for cancer and infectious diseases

    NASA Astrophysics Data System (ADS)

    Valentini, Paola; Persano, Stefano; Cecere, Paola; Sabella, Stefania; Pompa, Pier Paolo

    2014-03-01

    Traditional in vitro diagnostics requires specialized laboratories and costly instrumentation, both for the amplification of nucleic acid targets (usually achieved by PCR) and for the assay readout, often based on fluorescence. We are developing hybrid nanomaterials-based sensors for the rapid and low-cost diagnosis of various disease biomarkers, for applications in portable platforms for diagnostics at the point-of-care. To this aim, we exploited the size and distancedependent optical properties of gold nanoparticles (AuNPs) to achieve colorimetric detection. Moreover, in order to avoid the complexity of thermal cycles associated to traditional PCR, the design of our systems includes signal amplification schemes, achieved by the use of enzymes (nucleases, helicase) or DNAzymes. Focused on instrument-free and sensitive detection, we carefully combined the intrinsic sensitivity by multivalency of functionalized AuNPs with isothermal and non-stringent enzyme-aided reaction conditions, controlled AuNPs aggregates, universal reporters and magnetic microparticles, the latter used both as a substrate and as a means for the colorimetric detection. We obtained simple and robust assays for the sensitive (pM range or better) naked-eye detection of cancer or infectious diseases (HPV, HCV) biomarkers, requiring no instrumentation except for a simple heating plate. Finally, we are also developing non-medical applications of these bio-nanosensors, such as in the development of on-field rapid tests for the detection of pollutants and other food and water contaminants.

  7. [Detection of viable metabolically active yeast cells using a colorimetric assay].

    PubMed

    Růzicka, F; Holá, V

    2008-02-01

    The increasing concern of yeasts able to form biofilm brings about the need for susceptibility testing of both planktonic and biofilm cells. Detection of viability or metabolic activity of yeast cells after exposure to antimicrobials plays a key role in the assessment of susceptibility testing results. Colorimetric assays based on the color change of the medium in the presence of metabolically active cells proved suitable for this purpose. In this study, the usability of a colorimetric assay with the resazurin redox indicator for monitoring the effect of yeast inoculum density on the reduction rate was tested. As correlation between the color change rate and inoculum density was observed, approximate quantification of viable cells was possible. The assay would be of relevance to antifungal susceptibility testing in both planktonic and biofilm yeasts. PMID:18318392

  8. Multiplexed Colorimetric Solid-Phase Extraction

    NASA Technical Reports Server (NTRS)

    Gazda, Daniel B.; Fritz, James S.; Porter, Marc D.

    2009-01-01

    Multiplexed colorimetric solid-phase extraction (MC-SPE) is an extension of colorimetric solid-phase extraction (C-SPE) an analytical platform that combines colorimetric reagents, solid phase extraction, and diffuse reflectance spectroscopy to quantify trace analytes in water. In CSPE, analytes are extracted and complexed on the surface of an extraction membrane impregnated with a colorimetric reagent. The analytes are then quantified directly on the membrane surface using a handheld diffuse reflectance spectrophotometer. Importantly, the use of solid-phase extraction membranes as the matrix for impregnation of the colorimetric reagents creates a concentration factor that enables the detection of low concentrations of analytes in small sample volumes. In extending C-SPE to a multiplexed format, a filter holder that incorporates discrete analysis channels and a jig that facilitates the concurrent operation of multiple sample syringes have been designed, enabling the simultaneous determination of multiple analytes. Separate, single analyte membranes, placed in a readout cartridge create unique, analyte-specific addresses at the exit of each channel. Following sample exposure, the diffuse reflectance spectrum of each address is collected serially and the Kubelka-Munk function is used to quantify each water quality parameter via calibration curves. In a demonstration, MC-SPE was used to measure the pH of a sample and quantitate Ag(I) and Ni(II).

  9. High-throughput and rapid quantification of lipids by nanoflow UPLC-ESI-MS/MS: application to the hepatic lipids of rabbits with nonalcoholic fatty liver disease.

    PubMed

    Byeon, Seul Kee; Lee, Jong Cheol; Chung, Bong Chul; Seo, Hong Seog; Moon, Myeong Hee

    2016-07-01

    A rapid and high-throughput quantification method (approximately 300 lipids within 20 min) was established using nanoflow ultrahigh-pressure liquid chromatography-tandem mass spectrometry (nUPLC-ESI-MS/MS) with selective reaction monitoring (SRM) and applied to the quantitative profiling of the hepatic lipids of rabbits with different metabolic conditions that stimulate the development of nonalcoholic fatty liver disease (NAFLD). Among the metabolic conditions of rabbits in this study [inflammation (I), high-cholesterol diet (HC), and high-cholesterol diet combined with inflammation (HCI)], significant perturbation in hepatic lipidome (>3-fold and p < 0.01) was observed in the HC and HCI groups, while no single lipid showed a significant change in group I. In addition, this study revealed a dramatic increase (>2-fold) in relatively high-abundant monohexosylceramides (MHCs), sphingomyelins (SMs), and triacylglycerols (TGs) in both the HC and HCI groups, especially in MHCs as all 11 MHCs increased by larger than 3- to 12-fold. As the levels of the relatively high-abundant lipids in the above classes increased, the total lipidome level of each class increased significantly by approximately 2-fold to 5-fold. Other classes of lipids also generally increased, which was likely induced by the increase in mitogenic and nonapoptotic MHCs and SMs, as they promote cell proliferation. On the other hand, a slight decrease in the level of apoptotic ceramides (Cers) was observed, which agreed with the general increase in total lipid level. As distinct changes in hepatic lipidome were observed from HC groups, this suggests that HC or HCI is highly associated with NAFLD but not inflammation alone itself. Graphical Abstract Schematic of lipidomic analysis from hepatic tissue using nanoflow LC-ESI-MS/MS and nUPLC-ESI-MS/MS. PMID:27178550

  10. Rapid quantification of Escherichia coli in food and media using bacteriophage T7 amplification and liquid chromatography-multiple reaction monitoring tandem mass spectrometry.

    PubMed

    Banu, Mazlina; Ng, Daniel; Zheng, Lu; Goh, Lin-Tang; Bi, Xuezhi; Ow, Dave Siak-Wei

    2014-12-20

    Conventional microbiological assays have been a valuable tool for specific enumeration of indicative bacteria of relevance to food and public health, but these culture-based methods are time-consuming and require tedious biochemical and morphological identification. In this work, we exploit the ability of bacteriophage T7 to specifically infect Escherichia coli and amplify nearly a 100-fold in 1–2 h. Bacteriophage amplification is integrated with liquid chromatography-multiple reaction monitoring tandem mass spectrometry (LC-MRM–MS/MS) for quantitation of phage-specific peptides. Heavy isotopic 15N labeled T7 is introduced as the inoculum phage and internal standard. Quantification is performed by determining the ratio of phage-specific peptides over the internal standard which value is proportional to E. coli numbers. A broad dynamic range of 6-log orders ranging from 3.0 × 10(3) to 3.0 × 10(9) CFU/ml is attained in LB, while between 4.1 × 10(4)–2.7 × 10(9) CFU/ml and 1.9 × 10(3)–3.0 × 10(7) CFU/ml was enumerated respectively in coconut water and apple juice. With this method, viable E. coli are quantified in 4 h with a detection limit of 3.0 × 10(3) CFU/ml, 4.1 × 10(4) CFU/ml and 1.9 × 10(3) CFU/ml in LB, coconut water and apple juice, respectively. This method has potential as a rapid tool for detection of fecal contamination during food bioprocessing and distribution to safeguard public health. PMID:25456056

  11. Rapid and sensitive screening and selective quantification of antibiotics in human urine by two-dimensional ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry.

    PubMed

    Wang, He-Xing; Wang, Bin; Zhou, Ying; Jiang, Qing-Wu

    2014-12-01

    A rapid and sensitive method for the screening and selective quantification of antibiotics in urine by two-dimensional ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was developed. This method allowed the injection of 200 μL urine extract. The 200-μL injection volume used in this method increased the absolute sensitivity for target antibiotics in solvent by an average 13.3 times, with a range from 8.4 to 28.5 times, compared with the 10-μL conventional injection volume. A 96-well solid phase extraction procedure was established to eliminate the contamination on the chromatographic column resulting from the large-volume injection and increase the throughput of sample preparation. Fourteen target antibiotics from six common categories (β-lactams, quinolones, tetracyclines, macrolides, sulfonamides, and chloramphenicols) were selected as model compounds, and a database containing an additional 74 antibiotics was compiled for posttarget screening. The limit of detection of the target antibiotics, defined as a signal-to-noise ratio of 3, ranged from 0.04 to 1.99 ng/mL. The mean interday recoveries ranged between 79.6 and 121.3 %, with a relative standard deviation from 2.9 to 18.3 % at three spiking levels of 20 ng/mL, 50 ng/mL, and 100 ng/mL. This method was successfully applied in 60 real urine samples from schoolchildren aged 8-11 years, and four target antibiotics (azithromycin, sulfadiazine, trimethoprim, and oxytetracycline) and two posttarget antibiotics (sulfadimidine and cefaclor) were found in the urine samples. This method can be used as a large-scale biomonitoring tool for exposure of the human population to antibiotics. PMID:25354887

  12. A Colorimetric Bioassay for Perchlorate

    NASA Astrophysics Data System (ADS)

    Heinnickel, M. L.; Smith, S.; Coates, J. D.

    2007-12-01

    Recognition of perchlorate (ClO4-) as a widespread contaminant across the United States and its potential adverse affects towards human health has motivated the EPA to place ClO4- on its contaminant candidate list for drinking water supplies. While a federal MCL has not yet been set, a recommended public health goal of 1 ppb (μg.L-1) was established by the US EPA in 2002. To date, methods of detection require use of sensitive ion chromatographic equipment that are expensive, time consuming, and require highly trained personnel for use. Our studies are focused on the development of a highly sensitive, simple, and robust colorimetric bioassay based on the primary enzyme involved in microbial ClO4- reduction, the perchlorate reductase (Pcr). A previously published assay used reduced methyl viologen (MV, the dye is reduced with sodium hydrosulfite) as an electron donor to demonstrate Pcr activity. The assay directly correlates the amount of MV oxidized with the amount of ClO4- reduced by assuming a transfer of four electrons. To test this assumption, we compared actual concentrations of MV oxidized to ClO4- reduced in this assay. ClO4- concentrations were determined using a Dionex ICS-500 ion chromatography system, while MV concentrations were determined using a standard curve generated at 578 nm. Comparisons between the two revealed that twelve molecules of MV were oxidized for each molecule of ClO4- reduced. The oxidation of these additional eight MV molecules is explained by the interaction of the dye with chlorite (the product of the Pcr reaction) and other contaminants that could be present in the enzyme prep. This unsettling result indicated this assay would be problematic for the detection of ClO4- in soil, which has many chemicals that could react with MV. To improve upon this assay, we have tried to reduce ClO4- using less reactive dyes and reductants. The reductants ascorbic acid, NADH, and dithiothreitol drive Pcr catalyzed ClO4- reduction, however, they

  13. Colorimetric Immunoassay for Detection of Tumor Markers

    PubMed Central

    Yin, Yongmei; Cao, Ya; Xu, Yuanyuan; Li, Genxi

    2010-01-01

    Tumor markers are substances, usually proteins, produced by the body in response to cancer growth, or by the cancer tissue itself. They can be detected in blood, urine, or tissue samples, and the discovery and detection of tumor markers may provide earlier diagnosis of cancer and improved therapeutic intervention. Colorimetric immunoassays for tumor marker detection have attracted considerable attention, due to their simplicity and high efficiency. The traditionally used colorimetric immunoassays for the detection of tumor markers are based on enzyme-linked immunosorbent assays, and the great achievement of nanotechnology has further opened opportunities for the development of such kind of immunoassays. This paper will summarize recent advances in the field of colorimetric immunoassays for detecting tumor markers, which is aimed to provide an overview in this field, as well as experimental guidance for the learner. PMID:21614193

  14. Sensitiveness of the colorimetric estimation of titanium

    USGS Publications Warehouse

    Wells, R.C.

    1911-01-01

    The accuracy of the colorimetric estimation of titanium is practically constant over concentrations ranging from the strongest down to those containing about 1.5 mg. TiO2 in 100 cc. The change in concentration required to produce a perceptible difference in intensity between two solutions, at favorable concentrations, was found to be about 6.5 per cent, which does not differ much from the results of others with chromium and copper solutions. With suitable precautions, such as comparing by substitution and taking the mean of several settings or of the two perceptibly different extremes, the accuracy of the colorimetric comparisons appears to be about 2 per cent.

  15. Development of a Tunable LED-Based Colorimetric Source

    PubMed Central

    Brown, Steven W.; Santana, Carlos; Eppeldauer, George P.

    2002-01-01

    A novel, spectrally tunable light-source utilizing light emitting diodes (LEDs) for radiometric, photometric, and colorimetric applications is described. The tunable source can simulate standard sources and can be used as a transfer source to propagate photometric and colorimetric scales from calibrated reference instruments to test artifacts with minimal increase in uncertainty. In this prototype source, 40 LEDs with 10 different spectral distributions were mounted onto an integrating sphere. A voltage-to-current control circuit was designed and implemented, enabling independent control of the current sent to each set of four LEDs. The LEDs have been characterized for stability and dependence on drive current. The prototype source demonstrates the feasibility of development of a spectrally tunable LED source using LEDs with up to 40 different spectral distributions. Simulations demonstrate that such a source would be able to approximate standard light-source distributions over the visible spectral range—from 380 nm to 780 nm—with deviations on the order of 2 %. The tunable LED source can also simulate spectral distributions of special sources such as discharge lamps and display monitors. With this tunable source, a test instrument can be rapidly calibrated against a variety of different source distributions tailored to the anticipated uses of the artifact. Target uncertainties for the calibration of test artifacts are less than 2 % in luminance and 0.002 in chromaticity for any source distribution.

  16. Reactive Arrays of Colorimetric Sensors for Metabolite and Steroid Identification.

    PubMed

    Batres, Gary; Jones, Talia; Johnke, Hannah; Wilson, Mark; Holmes, Andrea E; Sikich, Sharmin

    2014-12-31

    The work described herein examines a rapid mix-and-measure method called DETECHIP suitable for screening of steroids and metabolites. The addition of steroids and metabolites to reactive arrays of colorimetric sensors generated characteristic color "fingerprints" that were used to identify the analyte. A color analysis tool was used to identify the analyte pool that now includes biologically relevant analytes. The mix-and-measure arrays allowed the detection of disease metabolites, orotic acid and argininosuccinic acid; and the steroids androsterone, 1,4-androstadiene, testosterone, stanozolol, and estrone. The steroid 1,4-androstadiene was also detected by this method while dissolved in synthetic urine. Some of the steroids, such as androstadiene, stanozolol, and androsterone were co-dissolved with (2-hydroxypropyl)-β-cyclodextrin in order to increase solubility in aqueous buffered solutions. The colorimetric arrays do not intend to eliminate ELISA or mass spectroscopy based screening, but to possibly provide an alternative analytical detection method for steroids and metabolites. PMID:25019034

  17. Development of a new colorimetric assay for lipoxygenase activity.

    PubMed

    Lu, Weiqiang; Zhao, Xue; Xu, Zhongyu; Dong, Ningning; Zou, Shien; Shen, Xu; Huang, Jin

    2013-10-15

    Lipoxygenases (LOXs) are a family of non-heme iron-containing dioxygenases that catalyze the hydroperoxidation of lipids, containing a cis,cis-1,4-pentadiene structure. A rapid and reliable colorimetric assay for determination of the activity of three human functional lipoxygenase isoforms (5-lipoxygenase, platelet 12-lipoxygenase, and 15-lipoxygenase-1) is developed in this article. In the new assay, LOX-derived lipid hydroperoxides oxidize the ferrous ion (Fe²⁺) to the ferric ion (Fe³⁺), the latter of which binds with thiocyanate (SCN⁻) to generate a red ferrithiocyanate (FTC) complex. The absorbance of the FTC complex can be easily measured at 480 nm. Because 5-LOX can be stimulated by many cofactors, the effects of its cofactors (Ca²⁺, ATP, dithiothreitol, glutathione, L-α-phosphatidylcholine, and ethylenediaminetetraacetic acid) on the color development of the FTC complex are also determined. The assay is adaptive for purified LOXs and cell lysates containing active LOXs. We use the new colorimetric assay in a 96-well format to evaluate several well-known LOX inhibitors, the IC₅₀ values of which are in good agreement with previously reported data. The reliability and reproducibility of the assay make it useful for in vitro screening for inhibitors of LOXs and, therefore, should accelerate drug discovery for clinical application. PMID:23811155

  18. Rapid Identification and Quantification of Natural Antioxidants in the Seeds of Rhubarb from Different Habitats in China Using Accelerated Solvent Extraction and HPLC-DAD-ESI-MSn-DPPH Assay.

    PubMed

    Tan, Liang; Geng, Dan-dan; Hu, Feng-zu; Dong, Qi

    2016-01-01

    In this study, the 10 accessions of rhubarb seeds from different habitats in China were investigated. Lipids were removed using petroleum ether, and the effective components were then separated using accelerated solvent extraction with 80% aqueous methanol. An off-line 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical scavenging method was used as the marker to evaluate the total antioxidant capability of extracts. On-line high-performance liquid chromatography-diode-array detectors-electrospray ionization-tandem mass spectrometry (HPLC-DAD-ESI-MS(n)) and HPLC-DAD-DPPH assays were developed for rapid identification and quantification of individual free-radical scavengers in extracts of rhubarb seeds. Ten free-radical scavengers from methanolic extracts of the rhubarb seeds were screened, five of which were identified and quantitatively analyzed: epicatechin, myricetin, hyperoside, quercitrin and quercetin. All were identified in rhubarb seeds for the first time and can be regarded as the major potent antioxidants in rhubarb seeds due to representing most of the total free-radical scavenging activity. Preliminary analysis of structures was performed for another five antioxidants. Based on our validation results, the developed method can be used for rapid separation, convenient identification and quantification of the multiple antioxidative constituents in rhubarb seeds, featuring good quantification parameters, accuracy and precision. The results are important to clarify the material basis and therapeutic mechanism of rhubarb seeds. PMID:26206792

  19. Nucleic acid-coupled colorimetric analyte detectors

    DOEpatents

    Charych, Deborah H.; Jonas, Ulrich

    2001-01-01

    The present invention relates to methods and compositions for the direct detection of analytes and membrane conformational changes through the detection of color changes in biopolymeric materials. In particular, the present invention provide for the direct colorimetric detection of analytes using nucleic acid ligands at surfaces of polydiacetylene liposomes and related molecular layer systems.

  20. Visualizing Capsaicinoids: Colorimetric Analysis of Chili Peppers

    ERIC Educational Resources Information Center

    Thompson, Robert Q.; Chu, Christopher; Gent, Robin; Gould, Alexandra P.; Rios, Laura; Vertigan, Theresa M.

    2012-01-01

    A colorimetric method for total capsaicinoids in chili pepper ("Capsicum") fruit is described. The placental material of the pepper, containing 90% of the capsaicinoids, was physically separated from the colored materials in the pericarp and extracted twice with methanol, capturing 85% of the remaining capsaicinoids. The extract, evaporated and…

  1. Colorimetric determination of tobramycin in parenteral solutions.

    PubMed

    Das Gupta, V

    1988-06-01

    A colorimetric method based on a reaction between tobramycin and alkaline copper sulphate solution has been proposed to quantify tobramycin in injections. The excipients present and normal saline did not interfere with the assay procedure. A tobramycin sample which was decomposed using either sulphuric acid or sodium hydroxide solution indicated fairly good stability on both sides of the pH scale. PMID:3209627

  2. Colorimetric determination of o-phenylenediamine in water samples based on the formation of silver nanoparticles as a colorimetric probe

    NASA Astrophysics Data System (ADS)

    Li, Nan; Gu, Yu; Gao, Mengmeng; Wang, Zilu; Xiao, Deli; Li, Yun; Lin, Rui; He, Hua

    2015-04-01

    A simple, rapid and cost-effective method for visual colorimetric detection of o-phenylenediamine (OPD) based on the formation of silver nanoparticles (AgNPs) has been developed in this paper. Silver ions can be reduced to AgNPs by OPD in a few minutes, causing changes in absorption spectra and color of the reaction system. Therefore, colorimetric detection of OPD could be realized by a UV-vis spectrophotometer or even the naked eye. Results showed that the absorption intensity of AgNPs at 416 nm exhibited a good linear correlation (R2 = 0.998) with OPD concentration in the range from 10-6 to 8 × 10-5 mol L-1 and the detection limit (3 σ/S) was calculated to be 1.61 × 10-7 mol L-1. Furthermore, as low as 4 × 10-6 mol L-1 OPD can be visualized by the naked eye without the requirement of any complicated or expensive instruments. This proposed method has been successfully applied to determine OPD in water samples, and may provide an innovative platform in the development of sensors for guiding environmental monitoring in the future.

  3. A Wash-Free Homogeneous Colorimetric Immunoassay Method

    PubMed Central

    Liu, Huiqiao; Rong, Pengfei; Jia, Hongwei; Yang, Jie; Dong, Bo; Dong, Qiong; Yang, Cejun; Hu, Pengzhi; Wang, Wei; Liu, Haitao; Liu, Dingbin

    2016-01-01

    Rapid and convenient biosensing platforms could be beneficial to timely diagnosis and treatment of diseases in virtually any care settings. Sandwich immunoassays, the most commonly used methods for protein detection, often rely on expensive tags such as enzyme and tedious wash and incubation procedures operated by skilled labor. In this report, we revolutionized traditional sandwich immunoassays by providing a wash-free homogeneous colorimetric immunoassay method without requirement of any separation steps. The proposed strategy was realized by controlling the growth of gold nanoparticles (AuNPs) to mediate the interparticle spacing in the protein-AuNP oligomers. We have demonstrated the successful in vitro detection of cancer biomarker in serum samples from patients with high clinical sensitivity and specificity. PMID:26722373

  4. Paper-based tuberculosis diagnostic devices with colorimetric gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Tsai, Tsung-Ting; Shen, Shu-Wei; Cheng, Chao-Min; Chen, Chien-Fu

    2013-08-01

    A colorimetric sensing strategy employing gold nanoparticles and a paper assay platform has been developed for tuberculosis diagnosis. Unmodified gold nanoparticles and single-stranded detection oligonucleotides are used to achieve rapid diagnosis without complicated and time-consuming thiolated or other surface-modified probe preparation processes. To eliminate the use of sophisticated equipment for data analysis, the color variance for multiple detection results was simultaneously collected and concentrated on cellulose paper with the data readout transmitted for cloud computing via a smartphone. The results show that the 2.6 nM tuberculosis mycobacterium target sequences extracted from patients can easily be detected, and the turnaround time after the human DNA is extracted from clinical samples was approximately 1 h.

  5. Hybrid nanosensor for colorimetric and ultrasensitive detection of nuclease contaminations

    NASA Astrophysics Data System (ADS)

    Cecere, Paola; Valentini, Paola; Pompa, Pier Paolo

    2016-04-01

    Nucleases are ubiquitous enzymes that degrade DNA or RNA, thus they can prejudice the good outcome of molecular biology experiments involving nucleic acids. We propose a colorimetric test for the naked-eye detection of nuclease contaminations. The system uses an hybrid nanosensor, based on gold nanoparticles functionalized with DNA probes. Our assay is rapid, instrument-free, simple and low-cost. Moreover, it reaches sensitivity equal or better than those of commercial kits, and presents a lot of advantageous aspects. Therefore, it is very competitive, with a real market potential. This test will be relevant in routine process monitoring in scientific laboratories, and in quality control in clinical laboratories and industrial processes, allowing the simultaneous detection of nucleases with different substrate specificities and large-scale screening.

  6. Optimized enzymatic colorimetric assay for determination of hydrogen peroxide (H2O2) scavenging activity of plant extracts

    PubMed Central

    Fernando, Chamira Dilanka; Soysa, Preethi

    2015-01-01

    The classical method to determine hydrogen peroxide (H2O2) scavenging activity of plant extracts is evaluated by measuring the disappearance of H2O2 at a wavelength of 230 nm. Since this method suffers from the interference of phenolics having strong absorption in the UV region, a simple and rapid colorimetric assay was developed where plant extracts are introduced to H2O2, phenol and 4-aminoantipyrine reaction system in the presence of horseradish peroxidase (HRP). This reaction yields a quinoneimine chromogen which can be measured at 504 nm. Decrease in the colour intensity reflects the H2O2 scavenged by the plant material. • Optimum conditions determined for this assay were 30 min reaction time, 37 °C, pH 7, enzyme concentration of 1 U/ml and H2O2 concentration of 0.7 mM. The limit of detection (LOD) and limit of quantitation (LOQ) were 136 μM and 411 μM, respectively. • Half maximal effective concentration required to scavenge 50% of H2O2 in the system (EC50 value) calculated for several plant extracts and standard antioxidants resulted in coefficient of variance (CV%) of the EC50 values less than 3.0% and correlation coefficient values (R2) > 0.95 for all dose response curves obtained. • This method is convenient and very precise which is suitable for the rapid quantification of H2O2 scavenging ability of standard antioxidants and natural antioxidants present in plant extracts. PMID:26285798

  7. Colorimetric detection of Shewanella oneidensis based on immunomagnetic capture and bacterial intrinsic peroxidase activity

    NASA Astrophysics Data System (ADS)

    Wen, Junlin; Zhou, Shungui; Chen, Junhua

    2014-06-01

    Rapid detection and enumeration of target microorganisms is considered as a powerful tool for monitoring bioremediation process that typically involves cleaning up polluted environments with functional microbes. A novel colorimetric assay is presented based on immunomagnetic capture and bacterial intrinsic peroxidase activity for rapidly detecting Shewanella oneidensis, an important model organism for environmental bioremediation because of its remarkably diverse respiratory abilities. Analyte bacteria captured on the immunomagnetic beads provided a bacterial out-membrane peroxidase-amplified colorimetric readout of the immunorecognition event by oxidizing 3, 3', 5, 5'-tetramethylbenzidine (TMB) in the present of hydrogen peroxide. The high-efficiency of immunomagnetic capture and signal amplification of peroxidase activity offers an excellent detection performance with a wide dynamic range between 5.0 × 103 and 5.0 × 106 CFU/mL toward target cells. Furthermore, this method was demonstrated to be feasible in detecting S. oneidensis cells spiked in environmental samples. The proposed colorimetric assay shows promising environmental applications for rapid detection of target microorganisms.

  8. Colorimetric detection of Shewanella oneidensis based on immunomagnetic capture and bacterial intrinsic peroxidase activity

    PubMed Central

    Wen, Junlin; Zhou, Shungui; Chen, Junhua

    2014-01-01

    Rapid detection and enumeration of target microorganisms is considered as a powerful tool for monitoring bioremediation process that typically involves cleaning up polluted environments with functional microbes. A novel colorimetric assay is presented based on immunomagnetic capture and bacterial intrinsic peroxidase activity for rapidly detecting Shewanella oneidensis, an important model organism for environmental bioremediation because of its remarkably diverse respiratory abilities. Analyte bacteria captured on the immunomagnetic beads provided a bacterial out-membrane peroxidase-amplified colorimetric readout of the immunorecognition event by oxidizing 3, 3′, 5, 5′-tetramethylbenzidine (TMB) in the present of hydrogen peroxide. The high-efficiency of immunomagnetic capture and signal amplification of peroxidase activity offers an excellent detection performance with a wide dynamic range between 5.0 × 103 and 5.0 × 106 CFU/mL toward target cells. Furthermore, this method was demonstrated to be feasible in detecting S. oneidensis cells spiked in environmental samples. The proposed colorimetric assay shows promising environmental applications for rapid detection of target microorganisms. PMID:24898751

  9. An organic indicator functionalized graphene oxide nanocomposite-based colorimetric assay for the detection of sarcosine

    NASA Astrophysics Data System (ADS)

    Xue, Zhonghua; Yin, Bo; Wang, Hui; Li, Mengqian; Rao, Honghong; Liu, Xiuhui; Zhou, Xinbin; Lu, Xiaoquan

    2016-03-01

    Rapid detection of sarcosine is a key requirement for both diagnosis and treatment of disease. We report here a simple yet sensitive colorimetric nanocomposite platform for rapid detection of sarcosine in alkaline media. The approach exploited the benefits of a rapid color-producing reaction between an organic indicator, 1,2-naphthoquinone-4-sulphonic acid sodium salt (NQS), and the analyte of sarcosine species as well as the good catalytic ability of graphene oxide (GO) to the formation of highly colored products due to its good water dispersibility, extremely large surface area and facile surface modification. As a result, a NQS functionalized GO nanocomposite through π-π stacking has been demonstrated to be useful as a highly efficient catalyst system for the selective and sensitive colorimetric determination of sarcosine by providing a nanocomposite-amplified colorimetric response. Meanwhile, the strategy offered excellent selectivity toward sarcosine species against other amino acids as well as a satisfying detection limit of 0.73 μM. More importantly, by using an electrochemical method, a credible sensing mechanism of GO nanocomposite-based colorimetric platform for a special analyte determination can be easily verified and elucidated, which also provides an attractive alternative to conventional characterization strategies.Rapid detection of sarcosine is a key requirement for both diagnosis and treatment of disease. We report here a simple yet sensitive colorimetric nanocomposite platform for rapid detection of sarcosine in alkaline media. The approach exploited the benefits of a rapid color-producing reaction between an organic indicator, 1,2-naphthoquinone-4-sulphonic acid sodium salt (NQS), and the analyte of sarcosine species as well as the good catalytic ability of graphene oxide (GO) to the formation of highly colored products due to its good water dispersibility, extremely large surface area and facile surface modification. As a result, a NQS

  10. A gold nanoparticles-based colorimetric test to detect single nucleotide polymorphisms for improvement of personalized therapy of psoriasis

    NASA Astrophysics Data System (ADS)

    Marsella, Alessandra; Valentini, Paola; Tarantino, Paolo; Congedo, Maurizio; Pompa, Pier Paolo

    2016-04-01

    We report a simple, rapid and low-cost test, based on gold nanoparticles, for the naked-eye colorimetric detection of a signature of single nucleotide polymorphisms (SNPs) relevant for the personalized medicine of psoriasis patients. We validated the colorimetric assay on real-world DNA samples from a cohort of 30 psoriasis patients and we compared the results, in double-blind, with those obtained with two state-of-the-art instrumental techniques, namely reverse dot blotting and direct sequencing, finding 100% agreement. We demonstrated high accuracy, sensitivity and specificity of the colorimetric test that can be easily adapted for the genotypization of different SNPs, important for the pharmacogenomics of various diseases, and in other fields, such as food traceability and population structure analysis.

  11. Colorimetric characterization models based on colorimetric characteristics evaluation for active matrix organic light emitting diode panels.

    PubMed

    Gong, Rui; Xu, Haisong; Tong, Qingfen

    2012-10-20

    The colorimetric characterization of active matrix organic light emitting diode (AMOLED) panels suffers from their poor channel independence. Based on the colorimetric characteristics evaluation of channel independence and chromaticity constancy, an accurate colorimetric characterization method, namely, the polynomial compensation model (PC model) considering channel interactions was proposed for AMOLED panels. In this model, polynomial expressions are employed to calculate the relationship between the prediction errors of XYZ tristimulus values and the digital inputs to compensate the XYZ prediction errors of the conventional piecewise linear interpolation assuming the variable chromaticity coordinates (PLVC) model. The experimental results indicated that the proposed PC model outperformed other typical characterization models for the two tested AMOLED smart-phone displays and for the professional liquid crystal display monitor as well. PMID:23089779

  12. A one-tube multiplexed colorimetric strategy based on plasmonic nanoparticles combined with non-negative matrix factorization.

    PubMed

    Liu, Yizhen; Fang, Wei; Wu, Zitong; Zhou, Guohua; Yi, Wen; Zhou, Xiaodong; Shen, Aiguo; Hu, Jiming

    2014-10-01

    Herein, a one-tube colorimetric platform has been developed for the simultaneous determination of two analytes (DNA as model object) in one tube with picomolar sensitivity. SPR-active nanoparticles are used to encode reporter probes sensitive to oligonucleotides associated with hepatitis A virus Vall7 polyprotein gene (HVA) and hepatitis B virus surface-antigen gene (HVB) respectively and magnetic beads (MBs) serve as the removal tool. In this mixed nanoparticles based biosensor, the addition of target analytes could change the concentration of each nanoparticle, leading to different colors of the supernatant. The influence of spectral overlap has been eliminated by a non-negative matrix factorization (NMF). With the assistance of NMF, the limit of detection (LOD) can be determinated as pM level without amplification. On the whole, this nanosensor boasts the advantages of high sensitivity and low sample consumption. Simultaneous colorimetric detection and quantification of two molecules in one tube are demonstrated. PMID:25059165

  13. Loop-Mediated Isothermal Amplification (LAMP) for Rapid Detection and Quantification of Dehalococcoides Biomarker Genes in Commercial Reductive Dechlorinating Cultures KB-1 and SDC-9

    PubMed Central

    Kanitkar, Yogendra H.; Stedtfeld, Robert D.; Steffan, Robert J.; Hashsham, Syed A.

    2016-01-01

    Real-time quantitative PCR (qPCR) protocols specific to the reductive dehalogenase (RDase) genes vcrA, bvcA, and tceA are commonly used to quantify Dehalococcoides spp. in groundwater from chlorinated solvent-contaminated sites. In this study, loop-mediated isothermal amplification (LAMP) was developed as an alternative approach for the quantification of these genes. LAMP does not require a real-time thermal cycler (i.e., amplification is isothermal), allowing the method to be performed using less-expensive and potentially field-deployable detection devices. Six LAMP primers were designed for each of three RDase genes (vcrA, bvcA, and tceA) using Primer Explorer V4. The LAMP assays were compared to conventional qPCR approaches using plasmid standards, two commercially available bioaugmentation cultures, KB-1 and SDC-9 (both contain Dehalococcoides species). DNA was extracted over a growth cycle from KB-1 and SDC-9 cultures amended with trichloroethene and vinyl chloride, respectively. All three genes were quantified for KB-1, whereas only vcrA was quantified for SDC-9. A comparison of LAMP and qPCR using standard plasmids indicated that quantification results were similar over a large range of gene concentrations. In addition, the quantitative increase in gene concentrations over one growth cycle of KB-1 and SDC-9 using LAMP was comparable to that of qPCR. The developed LAMP assays for vcrA and tceA genes were validated by comparing quantification on the Gene-Z handheld platform and a real-time thermal cycler using DNA isolated from eight groundwater samples obtained from an SDC-9-bioaugmented site (Tulsa, OK). These assays will be particularly useful at sites subject to bioaugmentation with these two commonly used Dehalococcoides species-containing cultures. PMID:26746711

  14. Visual Screening and Colorimetric Determination of Clenbuterol and Ractopamine Using Unmodified Gold Nanoparticles as Probe.

    PubMed

    Luo, Yeli; Liu, Xin; Guo, Jiajia; Gao, Hanting; Li, Ying; Xu, Jingyue; Shen, Fei; Sun, Chunyan

    2016-01-01

    In this paper, a sensitive method for the colorimetric detection of clenbuterol and ractopamine using citrate-stabilized gold nanoparticles (AuNPs) as probe was developed. The concentration of clenbuterol and ractopamine could be determined with naked eyes or a UV-vis spectrometer. By optimizing the influence of NaHSO₄ and incubation time, clenbuterol could be detected in the linear range of 0.1-4 µg/mL with the detection limit of 0.0158 µg/mL, and ractopamine could be detected in the linear range of 1-9 µg/mL with the detection limit of 0.0229 µg/mL. The proposed method could be successfully applied to detect clenbuterol and ractopamine in pig urines by a simple pre-treatment with excellent recoveries. The proposed colorimetric assay exhibits good reproducibility and accuracy, providing a simple and rapid method for the analysis of clenbuterol and ractopamine. PMID:27398486

  15. Comparison of a gas chromatographic and colorimetric method for the determination of plasma paracetamol.

    PubMed

    Chambers, R E; Jones, K

    1976-07-01

    Plasma paracetamol levels have generally been determined either by gas chromatography (Stewart and Willis, 1975), which relies on complex equipment, or by spectrophotometry (Knepil, 1974), which can be time-consuming. The introduction by Glynn and Kendal (1975) of a simple colorimetric method based on the reaction of paracetamol with nitrous acid to give 2-nitro-4-acetamidophenol appears to have overcome these disadvantages, thereby providing a suitable procedure for the rapid measurement of plasma paracetamol in cases of overdose. The method was reported to be specific for paracetamol, no interference being caused either by the sulphate and glucuronide conjugates of paracetamol or by a large number of other commonly found drugs. This communication presents the results of a study in which plasma paracetamol levels determined by the colorimetric method were compared with those determined by an established gas chromatographic technique. PMID:952476

  16. Discrimination of honeys using colorimetric sensor arrays, sensory analysis and gas chromatography techniques.

    PubMed

    Tahir, Haroon Elrasheid; Xiaobo, Zou; Xiaowei, Huang; Jiyong, Shi; Mariod, Abdalbasit Adam

    2016-09-01

    Aroma profiles of six honey varieties of different botanical origins were investigated using colorimetric sensor array, gas chromatography-mass spectrometry (GC-MS) and descriptive sensory analysis. Fifty-eight aroma compounds were identified, including 2 norisoprenoids, 5 hydrocarbons, 4 terpenes, 6 phenols, 7 ketones, 9 acids, 12 aldehydes and 13 alcohols. Twenty abundant or active compounds were chosen as key compounds to characterize honey aroma. Discrimination of the honeys was subsequently implemented using multivariate analysis, including hierarchical clustering analysis (HCA) and principal component analysis (PCA). Honeys of the same botanical origin were grouped together in the PCA score plot and HCA dendrogram. SPME-GC/MS and colorimetric sensor array were able to discriminate the honeys effectively with the advantages of being rapid, simple and low-cost. Moreover, partial least squares regression (PLSR) was applied to indicate the relationship between sensory descriptors and aroma compounds. PMID:27041295

  17. Rapid quantification of a cleavable antibody-conjugated drug by liquid chromatography/tandem mass spectrometry with microwave-assisted enzymatic cleavage.

    PubMed

    Xu, Ling; Packer, Laura E; Qian, Mark G; Wu, Jing-Tao

    2016-09-01

    Antibody-drug conjugates (ADCs) play an increasingly important role for targeted cancer treatment. One class of ADCs has attracted particular interest in drug development. These ADCs employ a cleavable chemistry linkage for drugs and utilize the reduced interchain disulfide cysteine residues for conjugation. In this work, a novel bioanalytical method for the quantification of a cleavable antibody-conjugated drug in plasma was developed, qualified, and implemented. This novel method significantly improves throughput by combining a microwave-assisted, enzymatic cleavage of conjugated drugs from ADCs with a 96-well based sample preparation procedure to immunocapture ADCs in plasma. The released drug is subsequently quantified using a LC/MS/MS method. Our results represent a high-throughput, generic, and sensitive quantification method for antibody-conjugated microtubule inhibitors (such as MMAE) for preclinical PK/PD studies. The linear range of the standard curve for antibody conjugated drug (MMAE) was from 2.01 to 2010ng/mL with an excellent linearity (r(2)>0.997). The intra-run precision was below 8.14% and accuracy was from -7.71% to -1.08%. No matrix effect or carryover was observed for this method. This method was successfully used to measure the level of conjugated drug in a preclinical PK/PD study in mice. PMID:27281578

  18. Rapid and sensitive quantification of levoglucosan in aerosols by high-performance anion-exchange chromatography with positive electrospray ionization mass spectrometry (HPAEC-positive ESI-MS)

    NASA Astrophysics Data System (ADS)

    Asakawa, Daichi; Furuichi, Yuko; Yamamoto, Atsushi; Oku, Yuichiro; Funasaka, Kunihiro

    2015-12-01

    A convenient quantification method for underivatized levoglucosan, which is a tracer for biomass burning influenced particulate matter (PM), has been established using high-performance anion-exchange chromatography (HPAEC) coupled to positive electrospray ionization mass spectrometry ((+)ESI-MS). Levoglucosan was chromatographically separated from its isomers (mannosan and galactosan) and detected selectively with positive ESI-MS. Limits of detection and quantification for this method were 0.40 and 1.3 ng mL-1, respectively. A comparison of simultaneous measurements by this method and conventional derivatization gas chromatography/mass spectrometry showed a good linearity with a slope of 1.008 and a determination coefficient of 0.9932. The developed method was applied to ambient suspended particulate matter hourly collected by continuous particulate monitors at 10 stations. The hourly concentration of levoglucosan during August 9-11, 2011, was 1.7-918 ng m-3 and its distribution indicated the transportation of biomass burning aerosols of a forest fire. This is the first report of horizontal distribution of the hourly levoglucosan concentration in Japan.

  19. New two dimensional liquid-phase colorimetric assay based on old iodine-starch complexation for the naked-eye quantitative detection of analytes.

    PubMed

    Nie, Jinfang; Brown, Tom; Zhang, Yun

    2016-06-14

    A new type of quantitative point-of-care liquid-phase colorimetric assay (LPCA) termed as two dimensional (2D) LPCA is designed using old iodine-starch complexation reaction firstly discovered in 1814. The naked-eye 2D LPCA's utility is well demonstrated with the simple, low-cost, portable detection and quantification of model analytes in buffer and/or human serum samples without the use of external electronic readers. PMID:27074248

  20. QUANTIFICATION AND INTERPRETATION OF TOTAL PETROLEUM HYDROCARBONS IN SEDIMENT SAMPLES BY A GC/MS METHOD AND COMPARISON WITH EPA 418.1 AND A RAPID FIELD METHOD

    EPA Science Inventory

    ABSTRACT: Total Petroleum hydrocarbons (TPH) as a lumped parameter can be easily and rapidly measured or monitored. Despite interpretational problems, it has become an accepted regulatory benchmark used widely to evaluate the extent of petroleum product contamination. Three cu...

  1. A rapid method for the simultaneous quantification of the major tocopherols, carotenoids, free and esterified sterols in canola (Brassica napus) oil using normal phase liquid chromatography.

    PubMed

    Flakelar, Clare L; Prenzler, Paul D; Luckett, David J; Howitt, Julia A; Doran, Gregory

    2017-01-01

    A normal phase high performance liquid chromatography (HPLC) method was developed to simultaneously quantify several prominent bioactive compounds in canola oil vis. α-tocopherol, γ-tocopherol, δ-tocopherol, β-carotene, lutein, β-sitosterol, campesterol and brassicasterol. The use of sequential diode array detection (DAD) and tandem mass spectrometry (MS/MS) allowed direct injection of oils, diluted in hexane without derivatisation or saponification, greatly reducing sample preparation time, and permitting the quantification of both free sterols and intact sterol esters. Further advantages over existing methods included increased analytical selectivity, and a chromatographic run time substantially less than other reported normal phase methods. The HPLC-DAD-MS/MS method was applied to freshly extracted canola oil samples as well as commercially available canola, palm fruit, sunflower and olive oils. PMID:27507459

  2. Development and validation of a rapid and sensitive LC-ESI-MS/MS method for ondansetron quantification in human plasma and its application in comparative bioavailability study.

    PubMed

    Moreira, Roberto F; Salvadori, Myriam C; Azevedo, Cristina P; Oliveira-Silva, Diogo; Borges, Diego C; Moreno, Ronílson A; Sverdloff, Carlos E; Borges, Ney C

    2010-11-01

    The validation of a high throughput and specific method using a high-performance liquid chromatography coupled to electrospray (ES+) ionization tandem triple quadrupole mass spectrometric (LC-ESI-MS/MS) method for ondansetron quantification in human plasma is described. Human plasma samples were extracted by liquid-liquid extraction (LLE) using methyl tert-butyl ether and analyzed by LC-ESI-MS/MS. The limit of quantification was 0.2 ng/mL and the method was linear in the range 0.2-60 ng/mL. The intra-assay precisions ranged from 1.6 to 7.7%, while inter-assay precisions ranged from 2.1 to 5.1%. The intra-assay accuracies ranged from 97.5 to 108.2%, and the inter-assay accuracies ranged from 97.3 to 107.0%. The analytical method was applied to evaluate the relative bioavailability of two pharmaceutical formulations containing 8 mg of ondansetron each in 25 healthy volunteers using a randomized, two-period crossover design. The geometric mean and respective 90% confidence interval (CI) of ondansetron test/reference percent ratios were 90.15% (81.74-99.44%) for C(max) and 93.11% (83.01-104.43%) for AUC(₀-t). Based on the 90% confidence interval of the individual ratios (test formulation/reference formulation) for C(max) and AUC(₀-inf), it was concluded that the test formulation is bioequivalent to the reference one with respect to the rate and extent of absorption of ondansetron. PMID:20954214

  3. Development and validation of a reliable and rapid LC-MS/MS method for simultaneous quantification of sacubitril and valsartan in rat plasma and its application to a pharmacokinetic study.

    PubMed

    Chunduri, Raja Haranadha Babu; Dannana, Gowri Sankar

    2016-09-01

    A selective, sensitive and rapid liquid chromatographic method with electrospray ionization tandem mass spectrometric detection has been developed and validated for simultaneous quantification of sacubitril and valsartan in rat plasma using telmisartan as internal standard (IS). The analytes were extracted by deprotenization of 50 μL of plasma sample using 200 μL of acetonitrile. In a short chromatographic run of 1.50 min run time, separation was achieved on a Hypersil Gold C18 column using a mobile phase composed of 0.1% formic acid in Milli-Q water-0.1% formic acid in acetonitrile in gradient elution mode. The quantification of target compounds was performed in a positive electrospray ionization mode and multiple reaction monitoring. Response was a linear function of concentration in the ranges of 0.5-20,000 ng/mL for both analytes, with r(2)  > 0.9997. The intra- and inter-day precision and accuracy results were <15% and acceptable as per US Food and Drug Administration guidelines. Stability of compounds were established in a battery of stability studies, i.e. bench-top, autosampler and long-term storage stability as well as freeze-thaw cycles. The validated method can be used as a routine method to support pharmacokinetic studies. Copyright © 2016 John Wiley & Sons, Ltd. PMID:26876742

  4. Development of a rapid column-switching LC-MS/MS method for the quantification of THCCOOH and THCCOOH-glucuronide in whole blood for assessing cannabis consumption frequency.

    PubMed

    Hädener, Marianne; Weinmann, Wolfgang; Schürch, Stefan; König, Stefan

    2016-03-01

    The concentration of 11-nor-9-carboxy-Δ(9)-tetrahydrocannabinol (THCCOOH) in whole blood is used as a parameter for assessing the consumption behavior of cannabis consumers. The blood level of THCCOOH-glucuronide might provide additional information about the frequency of cannabis use. To verify this assumption, a column-switching liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the rapid and direct quantification of free and glucuronidated THCCOOH in human whole blood was newly developed. The method comprised protein precipitation, followed by injection of the processed sample onto a trapping column and subsequent gradient elution to an analytical column for separation and detection. The total LC run time was 4.5 min. Detection of the analytes was accomplished by electrospray ionization in positive ion mode and selected reaction monitoring using a triple-stage quadrupole mass spectrometer. The method was fully validated by evaluating the following parameters: linearity, lower limit of quantification, accuracy and imprecision, selectivity, extraction efficiency, matrix effect, carry-over, dilution integrity, analyte stability, and re-injection reproducibility. All acceptance criteria were analyzed and the predefined criteria met. Linearity ranged from 5.0 to 500 μg/L for both analytes. The method was successfully applied to whole blood samples from a large collective of cannabis consumers, demonstrating its applicability in the forensic field. PMID:26781107

  5. Rapid Quantification of Bacteria in Infected Root Canals Using Fluorescence Reagents and a Membrane Filter: A Pilot Study on Its Clinical Application to the Evaluation of the Outcomes of Endodontic Treatment

    PubMed Central

    Sato, Takuichi; Yamaki, Keiko; Ishida, Naoko; Shoji, Megumi; Sato, Emika; Abiko, Yuki; Hashimoto, Kazuhiro; Takeuchi, Yasuhisa; Matsuyama, Junko; Shimauchi, Hidetoshi; Takahashi, Nobuhiro

    2012-01-01

    Objective. The bacterial examination has been performed during the course of the root canal treatment. In the present pilot study, the new developed method, using fluorescence reagents and a membrane filter, was applied to the detection and quantification of bacteria in infected root canals, in order to evaluate the outcomes of the treatment. Methods. Six infected root canals with periapical lesions from 5 subjects were included. Informed consent was obtained from all subjects (age ranges, 23–79 years). Samples from infected root canals were collected at the beginning of the treatment (termed #25 First), the end of the first day of treatment (termed #55 First), and the next appointment day (termed #55 Second). Then, the bacterial count (CFU) was measured using fluorescence reagents (4′,6′-diamidino-2-phenylindole and propidium iodide) and the polycarbonate membrane filter by Bioplorer. Results. The mean ± SD of CFU in the sample of “#25 First” was (1.0 ± 1.4) × 105. As the root canal treatment progressed, the CFU decreased as 7.9 × 103 (#55 First) and 4.3 × 102 (#55 Second). Conclusion. In the present pilot study, rapid detection and quantification of bacteria in infected root canals were found to be successfully performed using fluorescence reagents and a membrane filter (Bioplorer analysis). PMID:22693506

  6. Reversible colorimetric probes for mercury sensing.

    PubMed

    Coronado, Eugenio; Galán-Mascarós, José R; Martí-Gastaldo, Carlos; Palomares, Emilio; Durrant, James R; Vilar, Ramón; Gratzel, M; Nazeeruddin, Md K

    2005-09-01

    The selectivity and sensitivity of two colorimetric sensors based on the ruthenium complexes N719 [bis(2,2'-bipyridyl-4,4'-dicarboxylate)ruthenium(II) bis(tetrabutylammonium) bis(thiocyanate)] and N749 [(2,2':6',2' '-terpyridine-4,4',4' '-tricarboxylate)ruthenium(II) tris(tetrabutylammonium) tris(isothiocyanate)] are described. It was found that mercury ions coordinate reversibly to the sulfur atom of the dyes' NCS groups. This interaction induces a color change in the dyes at submicromolar concentrations of mercury. Furthermore, the color change of these dyes is selective for mercury(II) when compared with other ions such as lead(II), cadmium(II), zinc(II), or iron(II). The detection limit for mercury(II) ions--using UV-vis spectroscopy--in homogeneous aqueous solutions is estimated to be approximately 20 ppb for N719 and approximately 150 ppb for N749. Moreover, the sensor molecules can be adsorbed onto high-surface-area mesoporous metal oxide films, allowing reversible heterogeneous sensing of mercury ions in aqueous solution. The results shown herein have important implications in the development of new reversible colorimetric sensors for the fast, easy, and selective detection and monitoring of mercuric ions in aqueous solutions. PMID:16131215

  7. Using the iPhone as a device for a rapid quantitative analysis of trinitrotoluene in soil.

    PubMed

    Choodum, Aree; Kanatharana, Proespichaya; Wongniramaikul, Worawit; Daeid, Niamh Nic

    2013-10-15

    Mobile 'smart' phones have become almost ubiquitous in society and are typically equipped with a high-resolution digital camera which can be used to produce an image very conveniently. In this study, the built-in digital camera of a smart phone (iPhone) was used to capture the results from a rapid quantitative colorimetric test for trinitrotoluene (TNT) in soil. The results were compared to those from a digital single-lens reflex (DSLR) camera. The colored product from the selective test for TNT was quantified using an innovative application of photography where the relationships between the Red Green Blue (RGB) values and the concentrations of colorimetric product were exploited. The iPhone showed itself to be capable of being used more conveniently than the DSLR while providing similar analytical results with increased sensitivity. The wide linear range and low detection limits achieved were comparable with those from spectrophotometric quantification methods. Low relative errors in the range of 0.4 to 6.3% were achieved in the analysis of control samples and 0.4-6.2% for spiked soil extracts with good precision (2.09-7.43% RSD) for the analysis over 4 days. The results demonstrate that the iPhone provides the potential to be used as an ideal novel platform for the development of a rapid on site semi quantitative field test for the analysis of explosives. PMID:24054571

  8. Colorimetric and ultrasensitive bioassay based on a dual-amplification system using aptamer and DNAzyme.

    PubMed

    Tang, Longhua; Liu, Yang; Ali, Md Monsur; Kang, Dong Ku; Zhao, Weian; Li, Jinghong

    2012-06-01

    Rapid detection of ultralow amount of biomarkers in a biologically complex mixture remains a major challenge. Herein, we report a novel aptamer-based protein detection assay that integrates two signal amplification processes, namely, polymerase-mediated rolling-circle amplification (RCA) and DNA enzyme-catalyzed colorimetric reaction. The target biomarker is captured in a sandwich assay by primary aptamer-functionalized microbeads (MBs) and a secondary aptamer that is connected to a RCA primer/circular template complex. RCA reaction, which amplifies the single biomarker binding events by a factor of hundreds to thousands (the first amplification) produces a long DNA molecule containing multiple DNAzyme units. The peroxidase-like DNAzyme catalyzes the oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (the second amplification), which generates a blue-green colorimetric signal. This new biosensing platform permits the ultrasensitive, label-free, colorimetric detection of biomarker in real time. Using platelet-derived growth factor B-chain (PDGF-BB) as a model system, we demonstrated that our assay can detect a protein marker specifically in a serum-containing medium, at a concentration as low as 0.2 pg/mL in ∼2 h, which rivals traditional assays such as ELISA. We anticipate this simple methodology for biomarker detection can find utility in point-of-care applications. PMID:22533853

  9. Optimization of Polydiacetylene-Coated Superparamagnetic Magnetite Biosensor for Colorimetric Detection of Biomarkers.

    PubMed

    Chan, Terence; Verma, Mohit S; Gu, Frank X

    2015-04-01

    Biosensors for point-of-care testing of critical illnesses are urgently needed, especially in many areas of poor healthcare infrastructure. Polydiacetylene-based sensors are ideal because of their unique colorimetric properties where blue to red color shifts can be observed with the naked eye. In this work, a colorimetric biosensor capable of simple, rapid magnetic separation is optimized, using horse IgG as a model antibody, to obtain higher sensitivity. Composed of a unique combination of polydiacetylene and superparamagnetic iron oxide, the biosensor is fabricated at varying ratios of polydiacetylene to demonstrate optimization of color responsiveness. At increasing polydiacetylene ratios, improved color responsiveness and aqueous dispersion are observed, but the magnetic separation efficiency starts to suffer. The optimal color response is obtained at 90 wt% polydiacetylene. In addition, a 50 times improved lower detection limit of 0.01 mg/mL horse IgG is achieved, a relevant biomarker concentration for diagnosing sepsis. This platform provides a promising colorimetric biosensor for point-of-care use. PMID:26353474

  10. Rapid quantification of conjugated and unconjugated bile acids and C27 precursors in dried blood spots and small volumes of serum.

    PubMed

    Janzen, N; Sander, S; Terhardt, M; Das, A M; Sass, J O; Kraetzner, R; Rosewich, H; Rosevich, H; Peter, M; Sander, J

    2010-06-01

    The aim of the study was to develop a method for fast and reliable diagnosis of peroxisomal diseases and to facilitate differential diagnosis of cholestatic hepatopathy. For the quantification of bile acids and their conjugates as well as C(27) precursors di- and trihydroxycholestanoic acid (DHCA, THCA), in small pediatric blood samples we combined HPLC separation on a reverse-phase C18 column with ESI-MS/MS analysis in the negative ion mode. Analysis was done with good precision (CV 3,7%-11.1%) and sufficient sensitivity (LOQ: 11-91 nmol/L) without derivatization. Complete analysis of 17 free and conjugated bile acids from dried blood spots and 10 microL serum samples, respectively, was performed within 12 min. Measurement of conjugated primary bile acids plus DHCA and THCA as well as ursodeoxycholic acid was done in 4.5 min. In blood spots of healthy newborns, conjugated primary bile acids were found in the range of 0.01 to 2.01 micromol/L. Concentrations of C(27) precursors were below the detection limit in normal controls. DHCA and THCA were specifically elevated in cases of peroxysomal defects and one Zellweger patient. PMID:20093478

  11. Rapid quantification of imidazolium-based ionic liquids by hydrophilic interaction liquid chromatography: Methodology and an investigation of the retention mechanisms.

    PubMed

    Hawkins, Cory A; Rud, Anna; Guthrie, Margaret L; Dietz, Mark L

    2015-06-26

    The separation of nine N,N'-dialkylimidazolium-based ionic liquids (ILs) by an isocratic hydrophilic interaction high-performance liquid chromatographic method using an unmodified silica column was investigated. The chosen analytical conditions using a 90:10 acetonitrile-ammonium formate buffer mobile phase on a high-purity, unmodified silica column were found to be efficient, robust, and sensitive for the determination of ILs in a variety of solutions. The retention window (k' = 2-11) was narrower than that of previous methods, resulting in a 7-min runtime for the nine IL homologues. The lower limit of quantification of the method, 2-3 μmol L(-1), was significantly lower than those reported previously for HPLC-UV methods. The effects of systematically modifying the IL cation alkyl chain length, column temperature, and mobile-phase water and buffer content on solute retention were examined. Cation exchange was identified as the dominant retention mechanism for most of the solutes, with a distinct (single methylene group) transition to a dominant partitioning mode at the highest solute polarity. PMID:25979537

  12. Rapid Quantification of Four Anthocyanins in Red Grape Wine by Hydrophilic Interaction Liquid Chromatography/Triple Quadrupole Linear Ion Trap Mass Spectrometry.

    PubMed

    Sun, Yongming; Xia, Biqi; Chen, Xiangzhun; Duanmu, Chuansong; Li, Denghao; Han, Chao

    2015-01-01

    The identification and quantification of four anthocyanins (cyanidin-3-O-glucoside, peonidin-3-O-glucoside, delphinidin-3-O-glucoside, and malvidin-3-O-glucoside) in red grape wine were carried out by hydrophilic interaction liquid chromatography/triple quadrupole linear ion trap MS (HILIC/QTrap-MS/MS). Samples were diluted directly and separated on a Merck ZIC HILIC column with 20 mM ammonium acetate solution-acetonitrile mobile phase. Quantitative data acquisition was carried out in the multiple reaction monitoring mode. Additional identification and confirmation of target compounds were performed using the enhanced product ion mode of the linear ion trap. The LOQs were in the range 0.05-1.0 ng/mL. The average recoveries were in the range 94.6 to 104.5%. The HILIC/QTrap-MS/MS platform offers the best sensitivity and specificity for characterization and quantitative determination of the four anthocyanins in red grape wines and fulfills the quality criteria for routine laboratory application. PMID:26651575

  13. Beetroot-pigment-derived colorimetric sensor for detection of calcium dipicolinate in bacterial spores.

    PubMed

    Gonçalves, Letícia Christina Pires; Da Silva, Sandra Maria; DeRose, Paul C; Ando, Rômulo Augusto; Bastos, Erick Leite

    2013-01-01

    In this proof-of-concept study, we describe the use of the main red beet pigment betanin for the quantification of calcium dipicolinate in bacterial spores, including Bacillus anthracis. In the presence of europium(III) ions, betanin is converted to a water-soluble, non-luminescent orange 1∶1 complex with a stability constant of 1.4 × 10(5) L mol(-1). The addition of calcium dipicolinate, largely found in bacterial spores, changes the color of the aqueous solution of [Eu(Bn)(+)] from orange to magenta. The limit of detection (LOD) of calcium dipicolinate is around 2.0 × 10(-6) mol L(-1) and the LOD determined for both spores, B. cereus and B. anthracis, is (1.1 ± 0.3)× 10(6) spores mL(-1). This simple, green, fast and low cost colorimetric assay was selective for calcium dipicolinate when compared to several analogous compounds. The importance of this work relies on the potential use of betalains, raw natural pigments, as colorimetric sensors for biological applications. PMID:24019934

  14. Development of an extraction method based on new porous organogel materials coupled with liquid chromatography-mass spectrometry for the rapid quantification of bisphenol A in urine.

    PubMed

    ter Halle, Alexandra; Claparols, Catherine; Garrigues, Jean Christophe; Franceschi-Messant, Sophie; Perez, Emile

    2015-10-01

    A new method based on the use of porous organogel materials in combination with liquid chromatography-tandem mass spectrometry (LC-MS-MS) was assessed for the quantification of trace contaminants in complex matrices. As a demonstration of the use of these new materials, the contaminant chosen as a model was bisphenol A (BPA) and its extraction was investigated in urine. Organogel materials consist of an organic solvent immobilized by an organogelator. The composition of the organogel materials was optimized in terms of extraction efficiency and compatibility with LC-MS-MS. Porosity was introduced into the organogel by means of the particulate leaching method using sugar crystals. This new absorbing material is simple to use; the extraction method is reduced to a few steps. The originality of the method lies in the complete dissolution of the material for analysis by LC-MS-MS. The matrix effect of the organogel components was studied and was found to be minimal in atmospheric-pressure chemical ionization (APCI) compared to electrospray ionization (ESI) in negative mode. The influence of matrix components on the extraction was investigated by working with different media (acidified water, synthetic urine, horse urine and human urine). The partition coefficient was not affected within the margin of error (±0.1). After optimization, bisphenol A recoveries from urine samples reached 80%. The actual concentration factor was 10. The relative standard deviation (RSD, n=6) for the extraction and determination of BPA in horse urine spiked at 10ngmL(-1) was 9%. Tests with spiked human urine showed that the extraction performances were the same as with the solutions tested previously. The use of porous organogel allowed a fast, simple, sensitive, robust, green method to be developed for the determination of trace contaminants in complex matrices. PMID:26342874

  15. Development of a rapid HPLC-UV method for simultaneous quantification of protodioscin and rutin in white and green asparagus spears.

    PubMed

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

    2010-01-01

    Asparagus (Asparagus officinalis L.) spears are rich in bioactive compounds such as protodioscin, a saponin, and rutin, a flavonoid. Protodioscin and rutin are routinely quantified separately, and an approach permitting simultaneous measurement would significantly improve speed of analysis. We have optimized an extraction procedure and modified a method of high-performance liquid chromatography by coupling to an ultraviolet detector to simultaneously analyze protodioscin and rutin in asparagus extracts. An acidic ethanol solvent was more efficient than methanol, acetonitrile, or water in coextraction of protodioscin and rutin. Protodioscin and rutin were detected at 210 nm, with retention times of 12.6 min and 7.9 min, respectively. The method was validated by high linear correlations between 3.13 and 1000.0 μg/mL for protodioscin (r(2)= 0.9999), and between 0.3 and 1087.5 μg/mL for rutin (r(2)= 0.9997). The limit(s) of detection and quantification for protodioscin were 1.6 μg/mL and 3.13 μg/mL, respectively, and for rutin 0.2 μg/mL and 0.3 μg/mL, respectively. White asparagus spears and the crown of the plants were revealed to be rich sources of protodioscin and contained 2.59 to 10.4 mg/g dry weight. Green asparagus spears, particularly the upper portion, were rich in rutin and contained between 1.51 and 7.29 mg/g dry weight. PMID:21535581

  16. Colorimetric calibration of coupled infrared simulation system

    NASA Astrophysics Data System (ADS)

    Zhang, Ying; Fei, Jindong; Gao, Yang; Du, Jian

    2015-10-01

    In order to test 2-color infrared sensors, a coupled infrared simulation system can generate radiometric outputs with wavelengths that range from less than 3 microns to more than 12 microns. There are two channels in the coupled simulation system, optically combined by a diachronic beam combiner. Each channel has an infrared blackbody, a filter, a diaphragm, and diaphragm-motors. The system is projected to the sensor under testing by a collimator. This makes it difficult to calibrate the system with only one-band thermal imager. Errors will be caused in the radiance levels measured by the narrow band thermal imager. This paper describes colorimetric temperature measurement techniques that have been developed to perform radiometric calibrations of these infrared simulation systems above. The calibration system consists of two infrared thermal imagers; one is operated at the wavelength range of MW-IR, and the other at the range of LW-IR.

  17. Sensitive colorimetric cytotoxicity measurement using alarmar blue.

    PubMed

    Page, B; Page, M

    1995-01-01

    Cytotoxicity testing of anticancer drugs requires techniques which are sensitive, reproductible and applicable to large scale testing using automated instruments. End point staining of cells or viability stains are reliable but they are often not sensitive enough or not suitable for automation. We have already described a fluorometric cytotoxicity assay using a non fluorescent substrate, Alarmar Blue, which upon reduction in living cells, yields a fluorescent product. This assay is sensitive and highly reproductible but it is limited since automated fluorescent plate readers are not found frequently in laboratories. The conditions are now described for using Alamar Blue as a substrate in a colorimetric cytotoxicity assay. This new economical assay could be used with advantage for large scale screening of cytotoxic agents in microtitration plates. PMID:21597689

  18. Accurate colorimetric feedback for RGB LED clusters

    NASA Astrophysics Data System (ADS)

    Man, Kwong; Ashdown, Ian

    2006-08-01

    We present an empirical model of LED emission spectra that is applicable to both InGaN and AlInGaP high-flux LEDs, and which accurately predicts their relative spectral power distributions over a wide range of LED junction temperatures. We further demonstrate with laboratory measurements that changes in LED spectral power distribution with temperature can be accurately predicted with first- or second-order equations. This provides the basis for a real-time colorimetric feedback system for RGB LED clusters that can maintain the chromaticity of white light at constant intensity to within +/-0.003 Δuv over a range of 45 degrees Celsius, and to within 0.01 Δuv when dimmed over an intensity range of 10:1.

  19. A novel coumarin Schiff-base as a Ni(II) ion colorimetric sensor.

    PubMed

    Wang, Lingyun; Ye, Decheng; Cao, Derong

    2012-05-01

    A novel coumarin Schiff base compound (L) prepared from 7-diethylaminocoumarin-3-aldehyde and 3-amino-7-hydroxycoumarin was synthesized and evaluated as a chemoselective Ni(2+) sensor. Addition of Ni(2+) to CH(3)CN solution of L resulted in a rapid color change from yellow to red together with a large red shift from 465 to 516 nm. Moreover, other common alkali-, alkaline earth-, transition- and rare earth metal ions induced no or minimal spectral changes. Experimental results indicated that L could be used as a potential Ni(2+) colorimetric and naked-eye chemosensor in CH(3)CN solution. PMID:22306449

  20. Colorimetric assay of noramidopyrine methanesulfonate sodium in formulations and in blood and urine samples.

    PubMed

    Diab, A H

    1977-04-01

    A simple, rapid, specific and sensitive colorimetric method is proposed for the quantitative estimation of noramido-pyrine methanesulfonate sodium in different dosage forms as well as in blood and urine samples. The method is based on the reaction of 3-sulfonic-5-amino-alpha-naphthol with formaldehyde liberated from noramidopyrine methanesulfonate sodium after treatment with conc. sulfuric acid where a yellow colour appeared immediately which turned to blue on dilution with water. The blue colour obeyed Beer's law (10--400 microgram) and remained stable for more than 1 h. The effect of other drugs, tablet excipients, parentral vehicles and suppository bases was studied. PMID:896911

  1. Chloride accelerated Fenton chemistry for the ultrasensitive and selective colorimetric detection of copper.

    PubMed

    Shan, Zhi; Lu, Mingsheng; Wang, Li; MacDonald, Bruce; MacInnis, Judy; Mkandawire, Martin; Zhang, Xu; Oakes, Ken D

    2016-02-01

    A highly selective, ultrasensitive (visual and instrumental detection limits of 40 nM and 0.1 nM, respectively), environmentally-friendly, simple and rapid colorimetric sensor was developed for the detection of copper(II) in water. This sensor is based on a novel signal-amplification mechanism involving reactive halide species (RHSs) including chlorides or bromides, which accelerate copper Fenton reactions oxidizing the chromogenic substrate to develop colour. The results of this study expand our understanding of copper-based Fenton chemistry. PMID:26685747

  2. Hemicyanine-based colorimetric chemosensors: Different recognition mechanisms for CN- sensing

    NASA Astrophysics Data System (ADS)

    Gwon, Seon-Yeong; Lee, Eun-Mi; Kim, Sung-Hoon

    2012-10-01

    Simple hemicyanine dyes were synthesized by a classical condensation reaction. The structures of these dyes were characterized by 1H NMR spectroscopy, elemental analysis, and FAB-Mass spectrometry. The sensing behavior of the hemicyanines towards a selection of anions was investigated by UV-vis absorption spectroscopy. These structurally simple dyes displayed a rapid response and high selectivity for cyanide ion over other common anions in the DMSO/H2O solution. Different sensing mechanism of the dyes to cyanide ion were confirmed by 1H NMR studies, together with theoretical calculations based on DFT and PPP-MO methods. These dyes show no colorimetric response for other anions investigated.

  3. Rapid separation and concentration of food-borne pathogens in food samples prior to quantification by viable-cell counting and real-time PCR.

    PubMed

    Fukushima, Hiroshi; Katsube, Kazunori; Hata, Yukiko; Kishi, Ryoko; Fujiwara, Satomi

    2007-01-01

    Buoyant density gradient centrifugation has been used to separate bacteria from complex food matrices, as well as to remove compounds that inhibit rapid detection methods, such as PCR, and to prevent false-positive results due to DNA originating from dead cells. Applying a principle of buoyant density gradient centrifugation, we developed a method for rapid separation and concentration following filtration and low- and high-speed centrifugation, as well as flotation and sedimentation buoyant density centrifugation, for 12 food-borne pathogens (Salmonella enterica, Escherichia coli, Yersinia enterocolitica, Campylobacter jejuni, Vibrio cholerae O139, Vibrio parahaemolyticus O3K6, Vibrio vulnificus, Providencia alcalifaciens, Aeromonas hydrophila, Bacillus cereus, Staphylococcus aureus, and Clostridium perfringens) in 13 different food homogenates. This method can be used prior to real-time quantitative PCR (RTi-qPCR) and viable-cell counting. Using this combined method, the target organisms in the food samples theoretically could be concentrated 250-fold and detected at cell concentrations as low as 10(1) to 10(3) CFU/g using the RTi-qPCR assay, and amounts as small as 10(0) to 10(1) CFU/g could be isolated using plate counting. The combined separation and concentration methods and RTi-qPCR confirmed within 3 h the presence of 10(1) to 10(2) CFU/g of Salmonella and C. jejuni directly in naturally contaminated chicken and the presence of S. aureus directly in remaining food items in a poisoning outbreak. These results illustrated the feasibility of using these assays for rapid inspection of bacterial food contamination during a real-world outbreak. PMID:17056684

  4. Multi-class, multi-residue analysis of trace veterinary drugs in milk by rapid screening and quantification using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry.

    PubMed

    Zhang, Yaqian; Li, Xiang; Liu, Xiaomao; Zhang, Jinjie; Cao, Yanzhong; Shi, Zhihong; Sun, Hanwen

    2015-12-01

    A simple and rapid multi-class multi-residue analytical method was developed for the screening and quantification of veterinary drugs in milk by ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). A total of 90 veterinary drugs investigated belonged to almost 20 classes including lincomycins, macrolides, sulfonamides, quinolones, tetracyclines, β-agonists, β-lactams, sedatives, β-receptor antagonists, sex hormones, glucocorticoids, nitroimidazoles, benzimidazoles, nitrofurans, and some others. A modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) procedure was developed for the sample preparation without the solid-phase extraction step. The linearity, sensitivity, accuracy, repeatability, and reproducibility of the method were fully validated. The response of the detector was linear for each target compound in a wide concentration range with a correlation coefficient (R(2)) of 0.9973 to 0.9999 (among them R(2)>0.999 for 73 of 90 analytes). The range of the limit of quantification for these compounds in the milk ranged from 0.10 to 17.30μg/kg. The repeatability and reproducibility were in the range of 2.11 to 9.62% and 2.76 to 13.9%, respectively. The average recoveries ranged from 72.62 to 122.2% with the RSD (n=6) of 1.30 to 9.61% at 3 concentration levels. For the screening method, the data of the precursor and product ions of the target analytes were simultaneously acquired under the all ions MS/MS mode in a single run. An accurate mass database for the confirmation and identification of the target compounds was established. The applicability of the screening method was verified by applying to real milk samples. The proposed analytical method allows the identification and confirmation of the target veterinary drugs at trace levels employing quick analysis time. Certain veterinary drugs were detected in some cases. PMID:26506545

  5. Ferromagnetic particles as a rapid and robust sample preparation for the absolute quantification of seven eicosanoids in human plasma by UHPLC-MS/MS.

    PubMed

    Suhr, Anna Catharina; Bruegel, Mathias; Maier, Barbara; Holdt, Lesca Miriam; Kleinhempel, Alisa; Teupser, Daniel; Grimm, Stefanie H; Vogeser, Michael

    2016-06-01

    We used ferromagnetic particles as a novel technique to deproteinize plasma samples prior to quantitative UHPLC-MS/MS analysis of seven eicosanoids [thromboxane B2 (TXB2), prostaglandin E2 (PGE2), PGD2, 5-hydroxyeicosatetraenoic acid (5-HETE), 11-HETE, 12-HETE, arachidonic acid (AA)]. A combination of ferromagnetic particle enhanced deproteination and subsequent on-line solid phase extraction (on-line SPE) realized quick and convenient semi-automated sample preparation-in contrast to widely used manual SPE techniques which are rather laborious and therefore impede the investigation of AA metabolism in larger patient cohorts. Method evaluation was performed according to a protocol based on the EMA guideline for bioanalytical method validation, modified for endogenous compounds. Calibrators were prepared in ethanol. The calibration curves were found to be linear in a range of 0.1-80ngmL(-1) (TXB2, PGE2, PGD2), 0.05-40ngmL(-1) (5-HETE, 11-HETE), 0.5-400ngmL(-1) (12-HETE) and 25-9800ngmL(-1) (AA). Regarding all analytes and all quality controls, the resulting precision data (inter-assay 2.6 %-15.5 %; intra-assay 2.5 %-15.1 %, expressed as variation coefficient) as well as the accuracy results (inter-assay 93.3 %-125 %; intra-assay 91.7 %-114 %) were adequate. Further experiments addressing matrix effect, recovery and robustness, yielded also very satisfying results. As a proof of principle, the newly developed LC-MS/MS assay was employed to determine the capacity of AA metabolite release after whole blood stimulation in healthy blood donors. For this purpose, whole blood specimens of 5 healthy blood donors were analyzed at baseline and after a lipopolysaccharide (LPS) induced blood cell activation. In several baseline samples some eicosanoids levels were below the Lower Limit of Quantification. However, in the stimulated samples all chosen eicosanoids (except PGD2) could be quantified. These results, in context with those obtained in validation, demonstrate the

  6. Evaluation of a PCR/DNA Probe Colorimetric Membrane Assay for Identification of Campylobacter spp. in Human Stool Specimens

    PubMed Central

    Collins, Evelyn; Glennon, Maura; Hanley, Shirley; Murray, Anne-Marie; Cormican, Martin; Smith, Terry; Maher, Majella

    2001-01-01

    DNA was extracted from 50 human stool specimens using the QIAamp DNA stool minikit. PCR amplification was followed by post-PCR hybridization to DNA probes specific for the Campylobacter genus, Campylobacter jejuni, and Campylobacter coli in a colorimetric membrane assay. Thirty-two of 38 culture-positive specimens were PCR/DNA probe positive for C. jejuni. The assay is rapid and simple and can be applied to stool specimens for the detection of Campylobacter. PMID:11682549

  7. ELISA-PLA: A novel hybrid platform for the rapid, highly sensitive and specific quantification of proteins and post-translational modifications.

    PubMed

    Tong, Qing-He; Tao, Tao; Xie, Li-Qi; Lu, Hao-Jie

    2016-06-15

    Detection of low-abundance proteins and their post-translational modifications (PTMs) remains a great challenge. A conventional enzyme-linked immunosorbent assay (ELISA) is not sensitive enough to detect low-abundance PTMs and suffers from nonspecific detection. Herein, a rapid, highly sensitive and specific platform integrating ELISA with a proximity ligation assay (PLA), termed ELISA-PLA, was developed. Using ELISA-PLA, the specificity was improved by the simultaneous and proximate recognition of targets through multiple probes, and the sensitivity was significantly improved by rolling circle amplification (RCA). For GFP, the limit of detection (LOD) was decreased by two orders of magnitude compared to that of ELISA. Using site-specific phospho-antibody and pan-specific phospho-antibody, ELISA-PLA was successfully applied to quantify the phosphorylation dynamics of ERK1/2 and the overall tyrosine phosphorylation level of ERK1/2, respectively. ELISA-PLA was also used to quantify the O-GlcNAcylation of AKT, c-Fos, CREB and STAT3, which is faster and more sensitive than the conventional immunoprecipitation and western blotting (IP-WB) method. As a result, the sample consumption of ELISA-PLA was reduced 40-fold compared to IP-WB. Therefore, ELISA-PLA could be a promising platform for the rapid, sensitive and specific detection of proteins and PTMs. PMID:26866564

  8. Transformation of microcystins to 2-methyl-3-methoxy-4-phenylbutyric acid by room temperature ozone oxidation for rapid quantification of total microcystins.

    PubMed

    Zhang, L L; Yu, R P; Wang, L P; Wu, S F; Song, Q J

    2016-04-20

    Microcystins (MCs) are cyanobacterial hepatotoxins capable of accumulation into animal tissues. To determine the total microcystins in water, a novel analytical method, including ozonolysis, methylation of 2-methyl-3-methoxy-4-phenylbutyric acid (MMPB) with methylchloroformate (MCF) and gas chromatography mass spectrometry (GC-MS) detection was developed. The results show that MCs can be oxidized by ozone to produce MMPB at ambient temperature, proving ozonation is an effective, rapid and green method for the transformation of MCs to MMPB without secondary pollution. The oxidation conditions as well as the esterification process were optimized and, subsequently applied to analysis of environmental samples. The method shows wide linear range and high sensitivity with a detection limit of 0.34 μg L(-1). The established method was successfully applied to the analysis of microcystins in water samples. PMID:26975781

  9. Quantification of rifampicin in human plasma and cerebrospinal fluid by a highly sensitive and rapid liquid chromatographic–tandem mass spectrometric method

    PubMed Central

    Srivastava, Abhishek; Waterhouse, David; Ardrey, Alison; Ward, Stephen A.

    2012-01-01

    A highly sensitive and rapid liquid chromatography tandem mass spectrometry (LC–MS/MS) method has been developed to measure the levels of the antitubercular drug rifampicin (RIF) in human plasma and cerebrospinal fluid (CSF). The analyte and internal standard (IS) were isolated from plasma and CSF by a simple organic solvent based precipitation of proteins followed by centrifugation. Detection was carried out by electrospray positive ionization mass spectrometry in the multiple-reaction monitoring (MRM) mode. The assay was linear in the concentration range 25–6400 ng/mL with intra- and inter-day precision of <7% and <8%, respectively. The validated method was applied to the study of RIF pharmacokinetics in human CSF and plasma over 25 h period after a 10 mg/kg oral dose. PMID:22709606

  10. Ultrafast colorimetric determination of predominant protein structure evolution with gold nanoplasmonic particles

    NASA Astrophysics Data System (ADS)

    Kim, Hye Young; Choi, Inhee

    2016-01-01

    The intracellular and extracellular accumulation of disordered proteins and aggregated proteins occurs in many protein conformational diseases, such as aging-related neurodegeneration and alcoholic liver diseases. However, the conventional methods to study protein structural changes are limited for the rapid detection and monitoring of protein aggregation because of long incubation times (i.e., usually several days), complicated sample pretreatment steps, and expensive instrumentation. Here, we describe an ultrafast colorimetric method for the real-time monitoring of protein structure evolution and the determination of predominant structures via nanoparticle-assisted protein aggregation. During the aggregation process, nanoparticles act as nucleation cores, which form networks depending on the structures of the protein aggregates, and accelerate the kinetics of the protein aggregation. Simultaneously, these nanoparticles exhibit colorimetric responses according to their embedded shapes (e.g., fibrillar and amorphous) on the protein aggregates. We observed distinct spectral shifts and concomitant colorimetric responses of concentration- and type-dependent protein aggregation with the naked eye within a few minutes (<2 min) under acidic conditions. Moreover, the morphological transitions from small aggregates to larger aggregates of nanoparticle-assisted protein aggregates were visualized with dark-field microscope imaging, which show a similar trend with that of protein aggregates formed without the aid of nanoparticles. Finally we show that our proposed method can be utilized to screen the protein aggregation propensity under a variety of conditions such as different pH levels, high temperature, and chemicals. These findings suggest that the proposed method is an easy way to study the molecular biophysics of protein aggregation and to rapidly screen anti-aggregation drugs for protein conformational diseases.The intracellular and extracellular accumulation of

  11. Rapid quantification of the toxic alga Prymnesium parvum in natural samples by use of a specific monoclonal antibody and solid-phase cytometry.

    PubMed

    West, N J; Bacchieri, R; Hansen, G; Tomas, C; Lebaron, P; Moreau, H

    2006-01-01

    The increasing incidence of harmful algal blooms around the world and their associated health and economic effects require the development of methods to rapidly and accurately detect and enumerate the target species. Here we describe use of a solid-phase cytometer to detect and enumerate the toxic alga Prymnesium parvum in natural samples, using a specific monoclonal antibody and indirect immunofluorescence. The immunoglobulin G antibody 16E4 exhibited narrow specificity in that it recognized several P. parvum strains and a Prymnesium nemamethecum strain but it did not cross-react with P. parvum strains from Scandinavia or any other algal strains, including species of the closely related genus Chrysochromulina. Prymnesium sp. cells labeled with 16E4 were readily detected by the solid-phase cytometer because of the large fluorescence signal and the signal/noise ratio. Immunofluorescence detection and enumeration of cultured P. parvum cells preserved with different fixatives showed that the highest cell counts were obtained when cells were fixed with either glutaraldehyde or formaldehyde plus the cell protectant Pluronic F-68, whereas the use of formaldehyde alone resulted in significantly lower counts. Immunofluorescence labeling and analysis with the solid-phase cytometer of fixed natural samples from a bloom of P. parvum occurring in Lake Colorado in Texas gave cell counts that were close to those obtained by the traditional method of counting using light microscopy. These results show that a solid-phase cytometer can be used to rapidly enumerate natural P. parvum cells and that it could be used to detect other toxic algae, with an appropriate antibody or DNA probe. PMID:16391128

  12. Dual-Modal Colorimetric/Fluorescence Molecular Probe for Ratiometric Sensing of pH and Its Application.

    PubMed

    Wu, Luling; Li, Xiaolin; Huang, Chusen; Jia, Nengqin

    2016-08-16

    As traditional pH meters cannot work well for minute regions (such as subcellular organelles) and in harsh media, molecular pH-sensitive devices for monitoring pH changes in diverse local heterogeneous environments are urgently needed. Here, we report a new dual-modal colorimetric/fluorescence merocyanine-based molecular probe (CPH) for ratiometric sensing of pH. Compared with previously reported pH probes, CPH bearing the benzyl group at the nitrogen position of the indolium group and the phenol, which is used as the acceptor for proton, could respond to pH changes immediately through both the ratiometric fluorescence signal readout and naked-eye colorimetric observation. The sensing process was highly stable and reversible. Most importantly, the suitable pKa value (6.44) allows CPH to presumably accumulate in lysosomes and become a lysosome-target fluorescent probe. By using CPH, the intralysosomal pH fluctuation stimulated by antimalaria drug chloroquine was successfully tracked in live cells through the ratiometric fluorescence images. Additionally, CPH could be immobilized on test papers, which exhibited a rapid and reversible colorimetric response to acid/base vapor through the naked-eye colorimetric analysis. This proof-of-concept study presents the potential application of CPH as a molecular tool for monitoring intralysosomal pH fluctuation in live cells, as well as paves the way for developing the economic, reusable, and fast-response optical pH meters for colorimetric sensing acid/base vapor with direct naked-eye observation. PMID:27431089

  13. Simple and Rapid Determination of Ferulic Acid Levels in Food and Cosmetic Samples Using Paper-Based Platforms

    PubMed Central

    Tee-ngam, Prinjaporn; Nunant, Namthip; Rattanarat, Poomrat; Siangproh, Weena; Chailapakul, Orawon

    2013-01-01

    Ferulic acid is an important phenolic antioxidant found in or added to diet supplements, beverages, and cosmetic creams. Two designs of paper-based platforms for the fast, simple and inexpensive evaluation of ferulic acid contents in food and pharmaceutical cosmetics were evaluated. The first, a paper-based electrochemical device, was developed for ferulic acid detection in uncomplicated matrix samples and was created by the photolithographic method. The second, a paper-based colorimetric device was preceded by thin layer chromatography (TLC) for the separation and detection of ferulic acid in complex samples using a silica plate stationary phase and an 85:15:1 (v/v/v) chloroform: methanol: formic acid mobile phase. After separation, ferulic acid containing section of the TLC plate was attached onto the patterned paper containing the colorimetric reagent and eluted with ethanol. The resulting color change was photographed and quantitatively converted to intensity. Under the optimal conditions, the limit of detection of ferulic acid was found to be 1 ppm and 7 ppm (S/N = 3) for first and second designs, respectively, with good agreement with the standard HPLC-UV detection method. Therefore, these methods can be used for the simple, rapid, inexpensive and sensitive quantification of ferulic acid in a variety of samples. PMID:24077320

  14. Rapid quantification of bacteria and viruses in influent, settled water, activated sludge and effluent from a wastewater treatment plant using flow cytometry.

    PubMed

    Ma, Lili; Mao, Guannan; Liu, Jie; Yu, Hui; Gao, Guanghai; Wang, Yingying

    2013-01-01

    As microbiological parameters are important in monitoring the correct operation of wastewater treatment plants and controlling the microbiological quality of wastewater, the abundances of total bacteria (including intact and damaged bacterial cells) and total viruses in wastewater were investigated using a combination of ultrasonication and flow cytometry. The comparisons between flow cytometry (FCM) and other cultivation-independent methods (adenosine tri-phosphate (ATP) analysis for bacteria enumeration and epifluorescence microscopy (EFM) for virus enumeration) gave very similar patterns of microbial abundance changes, suggesting that FCM is suitable for targeting and obtaining reliable counts for bacteria and viruses in wastewater samples. The main experimental results obtained were: (1) effective removal of total bacteria in wastewater, with a decrease from an average concentration of 1.74 × 10(8)counts ml(-1) in raw wastewater to 3.91 × 10(6)counts ml(-1) in the effluent, (2) compared to influent raw wastewater, the average concentration of total viruses in the treated effluent (3.94 × 10(8)counts ml(-1)) exhibited no obvious changes, (3) the applied FCM approach is a rapid, easy, and convenient tool for understanding the microbial dynamics and monitoring microbiological quality in wastewater treatment processes. PMID:24185058

  15. Rapid and efficient method for the quantification of lychnopholide in rat plasma by liquid chromatography-tandem mass spectrometry for pharmacokinetic application.

    PubMed

    Lachi-Silva, Larissa; Sousa, João Paulo Barreto; Montanha, Maiara Camotti; Sy, Sherwin K B; Lopes, João Luis Callegari; Silva, Denise Brentan; Lopes, Norberto Peporine; Diniz, Andréa; Kimura, Elza

    2016-07-01

    Lychnopholide is a sesquiterpene lactone usually obtained from Lychnophora and Eremanthus species and has pharmacological activities that include anti-inflammatory and anti-tumor. Lychnopholide isolated from Eremanthus matogrossenssis was analyzed in this study. The aims of this study were to develop and validate an analytical methodology by LC-MS/MS and to quantify lychnopholide in rat plasma. Chromatographic separation was achieved on a C18 column using isocratic elution with the mobile phase consisting of methanol and water (containing 0.1% formic acid) at a flow rate of 0.4 mL/min. The detection was performed in multiple-reaction monitoring mode using electrospray ionization in positive mode. The method validation was performed in accordance with regulatory guidelines and the results met the acceptance criteria. The linear range of detection was 10-200 ng/mL (r > 0.9961). The intra- and inter-day assay variability were <6.2 and <11.7%, respectively. The extraction recovery was approximately 63% using liquid-liquid extraction with chloroform. Lychnopholide was detected in plasma up to 60 min after intravenous administration in rats. This rapid and sensitive method for the analysis of the sesquiterpene lactone lychnopholide in rat plasma can be applied to pharmacokinetic studies of this compound. Copyright © 2016 John Wiley & Sons, Ltd. PMID:26577838

  16. Establishing a TaqMan-Based Real-Time PCR Assay for the rapid detection and quantification of the newly emerged duck tembusu virus

    PubMed Central

    2011-01-01

    To establish an accurate, rapid, and a quantifiable method for the detection of the newly emerged duck Tembusu virus (DTMUV) that recently caused a widespread infectious disease in ducks in China, we developed a TaqMan-based real-time PCR assay by using E gene-specific primers and a TaqMan probe. This real-time PCR assay was 100 times more sensitive than the conventional PCR. The reproducibility and specificity of the real-time PCR assay were confirmed using plasmids containing E genes or RNAs and DNAs extracted from well-known viruses causing duck diseases. The reliability of this real-time PCR assay was confirmed in 19 of the 24 swab samples, 22 of the 24 tissue samples collected from experimentally infected ducks, as well as 15 of the 21 clinical samples collected from sick ducks since they were verified as DTMUV-positive. The results reveal that the newly established real-time PCR assay might be a useful diagnostic method for epidemiologically investigating and closely observing the newly emerged DTMUV. PMID:21978536

  17. Flow cytometry-assisted quantification of γH2AX expression has potential as a rapid high-throughput biodosimetry tool.

    PubMed

    Achel, Daniel G; Serafin, Antonio M; Akudugu, John M

    2016-08-01

    Large-scale radiological events require immediate and accurate estimates of doses received by victims, and possibly the first responders, to assist in treatment decisions. Although there are numerous efforts worldwide to develop biodosimetric tools to adequately handle triage needs during radiological incidents, such endeavours do not seem to actively involve sub-Saharan Africa which currently has a significant level of nuclear-related activity. To initiate a similar interest in Africa, ex vivo radiation-induced γH2AX expression in peripheral blood lymphocytes from fourteen healthy donors was assessed using flow cytometry. While the technique shows potential for use as a rapid high-throughput biodosimetric tool for radiation absorbed doses up to 5 Gy, significant inter-individual differences in γH2AX expression emerged. Also, female donors exhibited higher levels of γH2AX expression than their male counterparts. To address these shortcomings, gender-based in-house dose-response curves for γH2AX induction in lymphocytes 2, 4, and 6 h after X-ray irradiation are proposed for the South African population. The obtained results show that γH2AX is a good candidate biomarker for biodosimetry, but might need some refinement and validation through further studies involving a larger cohort of donors. PMID:27262315

  18. Quantification of Gas-Wall Partitioning in Teflon Environmental Chambers Using Rapid Bursts of Low-Volatility Oxidized Species Generated in Situ.

    PubMed

    Krechmer, Jordan E; Pagonis, Demetrios; Ziemann, Paul J; Jimenez, Jose L

    2016-06-01

    Partitioning of gas-phase organic compounds to the walls of Teflon environmental chambers is a recently reported phenomenon than can affect the yields of reaction products and secondary organic aerosol (SOA) measured in laboratory experiments. Reported time scales for reaching gas-wall partitioning (GWP) equilibrium (τGWE) differ by up to 3 orders of magnitude, however, leading to predicted effects that vary from substantial to negligible. A new technique is demonstrated here in which semi- and low-volatility oxidized organic compounds (saturation concentration c* < 100 μg m(-3)) were photochemically generated in rapid bursts in situ in an 8 m(3) environmental chamber, and then their decay in the absence of aerosol was measured using a high-resolution chemical ionization mass spectrometer (CIMS) equipped with an "inlet-less" NO3(-) ion source. Measured τGWE were 7-13 min (rel. std. dev. 33%) for all compounds. The fraction of each compound that partitioned to the walls at equilibrium follows absorptive partitioning theory with an equivalent wall mass concentration in the range 0.3-10 mg m(-3). Measurements using a CIMS equipped with a standard ion-molecule reaction region showed large biases due to the contact of compounds with walls. On the basis of these results, a set of parameters is proposed for modeling GWP in chamber experiments. PMID:27138683

  19. Quantification of forest carbon degradation in Nicaragua using RapidEye remote sensing data: El Cuá and Wiwili case studies

    NASA Astrophysics Data System (ADS)

    Argoty, F. N.; Cifuentes, M.; Imbach, P. A.; Vilchez, S.; Casanoves, F.; Ibrahim, M.; Vierling, L. A.

    2012-12-01

    Forest degradation and deforestation affect ecosystem function and climate regulation services such as carbon storage. Historically, Central America has been a deforestation and forest degradation hotspot. Wiwili and El Cuá municipalities in northern Nicaragua are no exception, where subsistence agriculture and cattle ranch expansion have driven deforestation and other wood extraction activities, leading to various levels of forest degradation. Reduction of Emissions from forest Degradation and Deforestation (REDD) projects are proposed as a tool to slow the degradation and loss of carbon stocks by restoring carbon to its natural levels in order to mitigate carbon dioxide emissions that cause global warming. REDD projects require baseline estimations of current carbon stocks and forest degradation status. We estimated carbon stocks across a forest degradation gradient based on common biophysical variables and commercially available (RapidEye) remote sensing data. We measured 80 temporary forest plots (50x20m) for aboveground biomass to sample a gradient of forest degradation at two municipalities (El Cuá and Wiwili) in northern Nicaragua. We measured biomass in trees (≥10 cm DBH), saplings (5-9.9 cm DBH), other growth forms (ferns, palms and woody vines), and large detritus (snags and downed wood). Biomass was estimated by a range of allometric models and a constant conversion factor (0.47) was applied to calculate aboveground carbon stocks. Remote sensing data from a RapidEye scene for 02/2010 provided data for 5 spectral bands and 19 vegetation indexes at 6 m spatial resolution. Precipitation, temperature, altitude, slope, canopy cover, and aspect were also used as input variables for carbon modeling. We tested linear mixed models, generalized additive mixed models and regression tree approaches to explain carbon stocks based on vegetation indexes and biophysical variables. Additionally, we grouped plots into low (17-168 Mg C ha-1), medium (168-302 Mg C ha-1

  20. Evaluation of a rapid method for the simultaneous quantification of ribavirin, sofosbuvir and its metabolite in rat plasma by UPLC-MS/MS.

    PubMed

    Shi, Xiaojun; Zhu, Dedong; Lou, Jie; Zhu, Bo; Hu, Ai-rong; Gan, Dongmei

    2015-10-01

    A rapid and sensitive ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the determination of ribavirin, sofosbuvir and its metabolite GS-331007 in rat plasma was established. The analytes and the internal standard (midazolam) were separated on an Acquity UPLC BEH C18 chromatography column (2.1mm×50mm, 1.7μm) using gradient elution with a mobile phase of acetonitrile and 0.1% formic acid in water at a flow rate of 0.4mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z 245.1→113.1 for ribavirin, m/z 530.3→243.1 for sofosbuvir, m/z 261.5→113.1 for GS-331007 and m/z 326.2→291.1 for midazolam (IS) using a positive electrospray ionization interface. The method was validated over a concentration range of 5-1000ng/mL for ribavirin, 10-2000ng/mL for sofosbuvir and 10-2000ng/mL for GS-331007. Total time for each chromatograph was 3.0min. The intra- and inter-day precision and accuracy of the quality control samples at low, medium, and high concentration levels exhibited relative standard deviations (RSD) <10.0% and the accuracy values ranged from -10.6% to 11.6%. The method was successfully applied to a pharmacokinetic study of ribavirin, sofosbuvir and GS-331007 in rats. PMID:26363369

  1. Development of a real-time PCR assay (SYBR Green I) for rapid identification and quantification of scyphomedusae Aurelia sp.1 planulae

    NASA Astrophysics Data System (ADS)

    Wang, Jianyan; Zhen, Yu; Mi, Tiezhu; Yu, Zhigang; Wang, Guoshan

    2015-07-01

    The complicated life cycle of Aurelia spp., comprising benthic asexually-reproducing polyps and sexually-reproducing medusae, makes it hard for researchers to identify and track them, especially for early stage individuals, such as planulae. To solve this problem, we developed a real-time PCR assay (SYBR Green I) to identify planulae in both cultured and natural seawater samples. Species-specific primers targeting Aurelia sp.1 mitochondrial 16S rDNA (mt 16S rDNA) regions were designed. Using a calibration curve constructed with plasmids containing the Aurelia sp.1 mt 16S rDNA fragment and a standard curve for planulae, the absolute number of mt 16S rDNA copies per planula was determined and from that the total number of planulae per sample was estimated. For the field samples, a 100-fold dilution of the sample DNA combined with a final concentration of 0.2 μg/μL BSA in the PCR reaction mixture was used to remove real-time PCR inhibitors. Samples collected in Jiaozhou Bay from July to September 2012 were subsequently analyzed using this assay. Peak Aurelia sp.1 planula abundance occurred in July 2012 at stations near Hongdao Island and Qingdao offshore; abundances were very low in August and September. The real-time PCR assay (SYBR Green I) developed here negates the need for traditional microscopic identification, which is laborious and time-consuming, and can detect and quantify jellyfish planulae in field plankton samples rapidly and specifically.

  2. Rapid, online quantification of H2S in JP-8 fuel reformate using near-infrared cavity-enhanced laser absorption spectroscopy.

    PubMed

    Dong, Feng; Junaedi, Christian; Roychoudhury, Subir; Gupta, Manish

    2011-06-01

    One of the key challenges in reforming military fuels for use with fuel cells is their high sulfur content, which can poison the fuel cell anodes. Sulfur-tolerant fuel reformers can convert this sulfur into H(2)S and then use a desulfurizing bed to remove it prior to the fuel cell. In order to optimize and verify this desulfurization process, a gas-phase sulfur analyzer is required to measure H(2)S at low concentrations (<1 ppm(v)) in the presence of other reforming gases (e.g., 25-30% H(2), 10-15% H(2)O, 15% CO, 5% CO(2), 35-40% N(2), and trace amounts of light hydrocarbons). In this work, we utilize near-infrared cavity-enhanced optical absorption spectroscopy (off-axis ICOS) to quantify H(2)S in a JP-8 fuel reformer product stream. The sensor provides rapid (2 s), highly precise (±0.1 ppm(v)) measurements of H(2)S in reformate gases over a wide dynamic range (0-1000 ppm(v)) with a low detection limit (3σ = ±0.09 ppm(v) in 1 s) and minimal cross-interferences from other present species. It simultaneously quantifies CO(2) (±0.2%), CH(4) (±150 ppm(v)), C(2)H(4) (±30 ppm(v)), and H(2)O (±300 ppm(v)) in the reformed gas for a better characterization of the fuel reforming process. Other potential applications of this technology include measurement of coal syngas and H(2)S in natural gas. By including additional near-infrared, distributive feedback diode lasers, the instrument can also be extended to other reformate species, including CO and H(2). PMID:21486070

  3. Basic design principles of colorimetric vision systems

    NASA Astrophysics Data System (ADS)

    Mumzhiu, Alex M.

    1998-10-01

    Color measurement is an important part of overall production quality control in textile, coating, plastics, food, paper and other industries. The color measurement instruments such as colorimeters and spectrophotometers, used for production quality control have many limitations. In many applications they cannot be used for a variety of reasons and have to be replaced with human operators. Machine vision has great potential for color measurement. The components for color machine vision systems, such as broadcast quality 3-CCD cameras, fast and inexpensive PCI frame grabbers, and sophisticated image processing software packages are available. However the machine vision industry has only started to approach the color domain. The few color machine vision systems on the market, produced by the largest machine vision manufacturers have very limited capabilities. A lack of understanding that a vision based color measurement system could fail if it ignores the basic principles of colorimetry is the main reason for the slow progress of color vision systems. the purpose of this paper is to clarify how color measurement principles have to be applied to vision systems and how the electro-optical design features of colorimeters have to be modified in order to implement them for vision systems. The subject of this presentation far exceeds the limitations of a journal paper so only the most important aspects will be discussed. An overview of the major areas of applications for colorimetric vision system will be discussed. Finally, the reasons why some customers are happy with their vision systems and some are not will be analyzed.

  4. Colorimetric Sensor Array for White Wine Tasting

    PubMed Central

    Chung, Soo; Park, Tu San; Park, Soo Hyun; Kim, Joon Yong; Park, Seongmin; Son, Daesik; Bae, Young Min; Cho, Seong In

    2015-01-01

    A colorimetric sensor array was developed to characterize and quantify the taste of white wines. A charge-coupled device (CCD) camera captured images of the sensor array from 23 different white wine samples, and the change in the R, G, B color components from the control were analyzed by principal component analysis. Additionally, high performance liquid chromatography (HPLC) was used to analyze the chemical components of each wine sample responsible for its taste. A two-dimensional score plot was created with 23 data points. It revealed clusters created from the same type of grape, and trends of sweetness, sourness, and astringency were mapped. An artificial neural network model was developed to predict the degree of sweetness, sourness, and astringency of the white wines. The coefficients of determination (R2) for the HPLC results and the sweetness, sourness, and astringency were 0.96, 0.95, and 0.83, respectively. This research could provide a simple and low-cost but sensitive taste prediction system, and, by helping consumer selection, will be able to have a positive effect on the wine industry. PMID:26213946

  5. Performance evaluation of a colorimetric hydrazine dosimeter

    NASA Astrophysics Data System (ADS)

    Brenner, Karen P.; Rose-Pehrsson, Susan L.

    1994-06-01

    A dosimeter for real-time, colorimetric detection of hydrazine in air has been developed. The passive badge consists of a dosimeter card containing a vanillin solution coated on a thin paper substrate. The active patch consists of a thick cellulose substrate coated with a vanillin solution. When placed in a plastic sample holder attached to a personnel pump, up to 5 L/min can be drawn through the active badge substrate. Through a condensation reaction, vanillin reacts with hydrazine to form a colored product that absorbs in the visible region. The hydrazone formed in the reaction is yellow; its intensity is proportional to the dose. When exposed passively to hydrazine, the experimental detection limit is less than 20 ppb-hrs. Extrapolated results indicate a detection limit of less than 5 ppb-hrs for long sampling periods. Actively sampling of hydrazine vapors gives an experimental detection limit of less than 100 ppb-L at a sample rate of 5 L/min. Relative humidity effects on badge response were minor. High humidity enhanced the color development on the vanillin badge; while low humidity had no effect on badge response. Interference testing of the dosimeters revealed a tobacco smoke interference. Preliminary shelf life tests indicated no decrease in sensitivity to hydrazine when stored at room temperature for 6 months.

  6. Analysis of the Precursor rRNA Fractions of Rapidly Growing Mycobacteria: Quantification by Methods That Include the Use of a Promoter (rrnA P1) as a Novel Standard†

    PubMed Central

    Menéndez, María del Carmen; Rebollo, María José; Núñez, María del Carmen; Cox, Robert A.; García, María Jesús

    2005-01-01

    Mycobacterial species are able to control rRNA production through variations in the number and strength of promoters controlling their rrn operons. Mycobacterium chelonae and M. fortuitum are members of the rapidly growing mycobacterial group. They carry a total of five promoters each, encoded, respectively, by one and two rrn operons per genome. Quantification of precursor rrn transcriptional products (pre-rrn) has allowed detection of different promoter usage during cell growth. Bacteria growing in several culture media with different nutrient contents were compared. Balanced to stationary phases were analyzed. Most promoters were found to be used at different levels depending on the stage of bacterial growth and the nutrient content of the culture medium. Some biological implications are discussed. Sequences of the several promoters showed motifs that could be correlated to their particular level of usage. A product corresponding to the first rrnA promoter in both species, namely, rrnA P1, was found to contribute at a low and near-constant level to pre-rRNA synthesis, regardless of the culture medium used and the stage of growth analyzed. This product was used as a standard to quantitate rRNA gene expression by real-time PCR when M. fortuitum infected macrophages. It was shown that this bacterium actively synthesizes rRNA during the course of infection and that one of its rrn operons is preferentially used under such conditions. PMID:15629925

  7. A rapid and sensitive LC-MS/MS method for quantification of quercetin-3-O-β-d-glucopyranosyl-7-O-β-d-gentiobioside in plasma and its application to a pharmacokinetic study.

    PubMed

    He, Xin; Tao, Guizhou; Gao, Hang; Li, Keyan; Zhang, Yazhuo; Sun, Limin; Zhang, Yingjie

    2016-09-01

    A rapid and sensitive LC-MS/MS method with good accuracy and precision was developed and validated for the pharmacokinetic study of quercetin-3-O-β-d-glucopyranosyl-7-O-β-d-gentiobioside (QGG) in Sprague-Dawley rats. Plasma samples were simply precipitated by methanol and then analyzed by LC-MS/MS. A Venusil® ASB C18 column (2.1 × 50 mm, i.d. 5 μm) was used for separation, with methanol-water (50:50, v/v) as the mobile phase at a flow rate of 300 μL/min. The optimized mass transition ion-pairs (m/z) for quantitation were 787.3/301.3 for QGG, and 725.3/293.3 for internal standard. The linear range was 7.32-1830 ng/mL with an average correlation coefficient of 0.9992, and the limit of quantification was 7.32 ng/mL. The intra- and inter-day precision and accuracy were less than ±15%. At low, medium and high quality control concentrations, the recovery and matrix effect of the analyte and IS were in the range of 89.06-92.43 and 88.58-97.62%, respectively. The method was applied for the pharmacokinetic study of QGG in Sprague-Dawley rats. Copyright © 2016 John Wiley & Sons, Ltd. PMID:26848536

  8. A rapid and reliable UPLC-MS/MS method for the identification and quantification of fourteen synthetic anti-diabetic drugs in adulterated Chinese proprietary medicines and dietary supplements.

    PubMed

    Li, Ning; Cui, Mei; Lu, Xiumei; Qin, Feng; Jiang, Kun; Li, Famei

    2010-11-01

    An ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for simultaneous qualitative and quantitative analysis of 14 synthetic anti-diabetic drugs in adulterated Chinese proprietary medicines (CPMs) and dietary supplements. The samples were prepared by ultrasonic extraction with methanol and separated on a C₁₈ column with mobile phase consisting of acetonitrile and water (both containing 0.10% formic acid). Gradient elution was applied with a flow rate of 0.20 mL/min. Two transitions from protonated molecules were monitored for each synthetic anti-diabetic drug in positive mode of electrospray ionization (ESI). The two transitions, the peak area ratio of the two transitions and the retention time were used for identification. The more intensive transition was used for quantification. The analysis time was 6 min per sample. Satisfactory linear relationships were estimated between the peak area and the concentration with correlation coefficients higher than 0.995. The limit of detection ranged from 0.03 to 5.45 ng/mL. The relative standard deviation of intra-day precision was below 7.6%, the RSD of inter-day precision was below 15% and the relative error of accuracy was between -10 and 7.8%. The proposed method is rapid, selective, reliable and was successfully applied to the analysis of 30 real samples of 22 CPMs and eight dietary supplements from the local market in China. PMID:20954219

  9. Rapid Detection and Quantification of RNA of Ebola and Marburg Viruses, Lassa Virus, Crimean-Congo Hemorrhagic Fever Virus, Rift Valley Fever Virus, Dengue Virus, and Yellow Fever Virus by Real-Time Reverse Transcription-PCR

    PubMed Central

    Drosten, Christian; Göttig, Stephan; Schilling, Stefan; Asper, Marcel; Panning, Marcus; Schmitz, Herbert; Günther, Stephan

    2002-01-01

    Viral hemorrhagic fevers (VHFs) are acute infections with high case fatality rates. Important VHF agents are Ebola and Marburg viruses (MBGV/EBOV), Lassa virus (LASV), Crimean-Congo hemorrhagic fever virus (CCHFV), Rift Valley fever virus (RVFV), dengue virus (DENV), and yellow fever virus (YFV). VHFs are clinically difficult to diagnose and to distinguish; a rapid and reliable laboratory diagnosis is required in suspected cases. We have established six one-step, real-time reverse transcription-PCR assays for these pathogens based on the Superscript reverse transcriptase-Platinum Taq polymerase enzyme mixture. Novel primers and/or 5′-nuclease detection probes were designed for RVFV, DENV, YFV, and CCHFV by using the latest DNA database entries. PCR products were detected in real time on a LightCycler instrument by using 5′-nuclease technology (RVFV, DENV, and YFV) or SybrGreen dye intercalation (MBGV/EBOV, LASV, and CCHFV). The inhibitory effect of SybrGreen on reverse transcription was overcome by initial immobilization of the dye in the reaction capillaries. Universal cycling conditions for SybrGreen and 5′-nuclease probe detection were established. Thus, up to three assays could be performed in parallel, facilitating rapid testing for several pathogens. All assays were thoroughly optimized and validated in terms of analytical sensitivity by using in vitro-transcribed RNA. The ≥95% detection limits as determined by probit regression analysis ranged from 1,545 to 2,835 viral genome equivalents/ml of serum (8.6 to 16 RNA copies per assay). The suitability of the assays was exemplified by detection and quantification of viral RNA in serum samples of VHF patients. PMID:12089242

  10. A selectively fluorescein-based colorimetric probe for detecting copper(II) ion.

    PubMed

    Zhang, Li; Zhang, Xianhong

    2014-12-10

    A novel fluorescein derivative 3-bromo-5-methylsalicylaldehyde fluorescein hydrazone (BMSFH) has been synthesized by reacting fluorescein hydrazide with 3-bromo-5-methylsalicylaldehyde and was developed as a new colorimetric probe for detection of Cu(2+). In the presence of Cu(2+) the BMSFH exhibits a rapid color change from colorless to yellow together with an obvious new band appeared at 502nm in the UV-vis absorption spectra. However, other common alkali-, alkaline earth-, transition- and rare earth metal ions induced no or minimal spectral changes. This change is attributed to BMSFH via coordination with Cu(2+) in a 1:1 stoichiometry and this binding to Cu(2+) is reversible, as indicated by the bleaching of the color when the Cu(2+) is extracted with EDTA. Experimental results indicate that the BMSFH can provide a rapid, selective and sensitive response to Cu(2+) with a linear dynamic range 3.0-330μmol/L and can be used as a potential Cu(2+) colorimetric probe in aqueous solution. PMID:24929315

  11. Simple colorimetric detection of doxycycline and oxytetracycline using unmodified gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Li, Jie; Fan, Shumin; Li, Zhigang; Xie, Yuanzhe; Wang, Rui; Ge, Baoyu; Wu, Jing; Wang, Ruiyong

    2014-08-01

    The interaction between tetracycline antibiotics and gold nanoparticles was studied. With citrate-coated gold nanoparticles as colorimetric probe, a simple and rapid detection method for doxycycline and oxytetracycline has been developed. This method relies on the distance-dependent optical properties of gold nanoparticles. In weakly acidic buffer medium, doxycycline and oxytetracycline could rapidly induce the aggregation of gold nanoparticles, resulting in red-to-blue (or purple) colour change. The experimental parameters were optimized with regard to pH, the concentration of the gold nanoparticles and the reaction time. Under optimal experimental conditions, the linear range of the colorimetric sensor for doxycycline/oxytetracycline was 0.06-0.66 and 0.59-8.85 μg mL-1, respectively. The corresponding limit of detection for doxycycline and oxytetracycline was 0.0086 and 0.0838 μg mL-1, respectively. This assay was sensitive, selective, simple and readily used to detect tetracycline antibiotics in food products.

  12. Dystrophin quantification

    PubMed Central

    Anthony, Karen; Arechavala-Gomeza, Virginia; Taylor, Laura E.; Vulin, Adeline; Kaminoh, Yuuki; Torelli, Silvia; Feng, Lucy; Janghra, Narinder; Bonne, Gisèle; Beuvin, Maud; Barresi, Rita; Henderson, Matt; Laval, Steven; Lourbakos, Afrodite; Campion, Giles; Straub, Volker; Voit, Thomas; Sewry, Caroline A.; Morgan, Jennifer E.; Flanigan, Kevin M.

    2014-01-01

    Objective: We formed a multi-institution collaboration in order to compare dystrophin quantification methods, reach a consensus on the most reliable method, and report its biological significance in the context of clinical trials. Methods: Five laboratories with expertise in dystrophin quantification performed a data-driven comparative analysis of a single reference set of normal and dystrophinopathy muscle biopsies using quantitative immunohistochemistry and Western blotting. We developed standardized protocols and assessed inter- and intralaboratory variability over a wide range of dystrophin expression levels. Results: Results from the different laboratories were highly concordant with minimal inter- and intralaboratory variability, particularly with quantitative immunohistochemistry. There was a good level of agreement between data generated by immunohistochemistry and Western blotting, although immunohistochemistry was more sensitive. Furthermore, mean dystrophin levels determined by alternative quantitative immunohistochemistry methods were highly comparable. Conclusions: Considering the biological function of dystrophin at the sarcolemma, our data indicate that the combined use of quantitative immunohistochemistry and Western blotting are reliable biochemical outcome measures for Duchenne muscular dystrophy clinical trials, and that standardized protocols can be comparable between competent laboratories. The methodology validated in our study will facilitate the development of experimental therapies focused on dystrophin production and their regulatory approval. PMID:25355828

  13. Colorimetric paper-based detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes from large volumes of agricultural water.

    PubMed

    Bisha, Bledar; Adkins, Jaclyn A; Jokerst, Jana C; Chandler, Jeffrey C; Pérez-Méndez, Alma; Coleman, Shannon M; Sbodio, Adrian O; Suslow, Trevor V; Danyluk, Michelle D; Henry, Charles S; Goodridge, Lawrence D

    2014-01-01

    This protocol describes rapid colorimetric detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes from large volumes (10 L) of agricultural waters. Here, water is filtered through sterile Modified Moore Swabs (MMS), which consist of a simple gauze filter enclosed in a plastic cartridge, to concentrate bacteria. Following filtration, non-selective or selective enrichments for the target bacteria are performed in the MMS. For colorimetric detection of the target bacteria, the enrichments are then assayed using paper-based analytical devices (µPADs) embedded with bacteria-indicative substrates. Each substrate reacts with target-indicative bacterial enzymes, generating colored products that can be detected visually (qualitative detection) on the µPAD. Alternatively, digital images of the reacted µPADs can be generated with common scanning or photographic devices and analyzed using ImageJ software, allowing for more objective and standardized interpretation of results. Although the biochemical screening procedures are designed to identify the aforementioned bacterial pathogens, in some cases enzymes produced by background microbiota or the degradation of the colorimetric substrates may produce a false positive. Therefore, confirmation using a more discriminatory diagnostic is needed. Nonetheless, this bacterial concentration and detection platform is inexpensive, sensitive (0.1 CFU/ml detection limit), easy to perform, and rapid (concentration, enrichment, and detection are performed within approximately 24 hr), justifying its use as an initial screening method for the microbiological quality of agricultural water. PMID:24962090

  14. Colorimetric Paper-based Detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes from Large Volumes of Agricultural Water

    PubMed Central

    Bisha, Bledar; Adkins, Jaclyn A.; Jokerst, Jana C.; Chandler, Jeffrey C.; Pérez-Méndez, Alma; Coleman, Shannon M.; Sbodio, Adrian O.; Suslow, Trevor V.; Danyluk, Michelle D.; Henry, Charles S.; Goodridge, Lawrence D.

    2014-01-01

    This protocol describes rapid colorimetric detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes from large volumes (10 L) of agricultural waters. Here, water is filtered through sterile Modified Moore Swabs (MMS), which consist of a simple gauze filter enclosed in a plastic cartridge, to concentrate bacteria. Following filtration, non-selective or selective enrichments for the target bacteria are performed in the MMS. For colorimetric detection of the target bacteria, the enrichments are then assayed using paper-based analytical devices (µPADs) embedded with bacteria-indicative substrates. Each substrate reacts with target-indicative bacterial enzymes, generating colored products that can be detected visually (qualitative detection) on the µPAD. Alternatively, digital images of the reacted µPADs can be generated with common scanning or photographic devices and analyzed using ImageJ software, allowing for more objective and standardized interpretation of results. Although the biochemical screening procedures are designed to identify the aforementioned bacterial pathogens, in some cases enzymes produced by background microbiota or the degradation of the colorimetric substrates may produce a false positive. Therefore, confirmation using a more discriminatory diagnostic is needed. Nonetheless, this bacterial concentration and detection platform is inexpensive, sensitive (0.1 CFU/ml detection limit), easy to perform, and rapid (concentration, enrichment, and detection are performed within approximately 24 hr), justifying its use as an initial screening method for the microbiological quality of agricultural water. PMID:24962090

  15. Quantification of plant chlorophyll content using Google Glass.

    PubMed

    Cortazar, Bingen; Koydemir, Hatice Ceylan; Tseng, Derek; Feng, Steve; Ozcan, Aydogan

    2015-04-01

    Measuring plant chlorophyll concentration is a well-known and commonly used method in agriculture and environmental applications for monitoring plant health, which also correlates with many other plant parameters including, e.g., carotenoids, nitrogen, maximum green fluorescence, etc. Direct chlorophyll measurement using chemical extraction is destructive, complex and time-consuming, which has led to the development of mobile optical readers, providing non-destructive but at the same time relatively expensive tools for evaluation of plant chlorophyll levels. Here we demonstrate accurate measurement of chlorophyll concentration in plant leaves using Google Glass and a custom-developed software application together with a cost-effective leaf holder and multi-spectral illuminator device. Two images, taken using Google Glass, of a leaf placed in our portable illuminator device under red and white (i.e., broadband) light-emitting-diode (LED) illumination are uploaded to our servers for remote digital processing and chlorophyll quantification, with results returned to the user in less than 10 seconds. Intensity measurements extracted from the uploaded images are mapped against gold-standard colorimetric measurements made through a commercially available reader to generate calibration curves for plant leaf chlorophyll concentration. Using five plant species to calibrate our system, we demonstrate that our approach can accurately and rapidly estimate chlorophyll concentration of fifteen different plant species under both indoor and outdoor lighting conditions. This Google Glass based chlorophyll measurement platform can display the results in spatiotemporal and tabular forms and would be highly useful for monitoring of plant health in environmental and agriculture related applications, including e.g., urban plant monitoring, indirect measurements of the effects of climate change, and as an early indicator for water, soil, and air quality degradation. PMID:25669673

  16. Quantification of Plant Chlorophyll Content Using Google Glass

    PubMed Central

    Cortazar, Bingen; Koydemir, Hatice Ceylan; Tseng, Derek; Feng, Steve; Ozcan, Aydogan

    2015-01-01

    Measuring plant chlorophyll concentration is a well-known and commonly used method in agriculture and environmental applications for monitoring plant health, which also correlates with many other plant parameters including, e.g., carotenoids, nitrogen, maximum green fluorescence, etc. Direct chlorophyll measurement using chemical extraction is destructive, complex and time-consuming, which has led to the development of mobile optical readers, providing non-destructive but at the same time relatively expensive tools for evaluation of plant chlorophyll levels. Here we demonstrate accurate measurement of chlorophyll concentration in plant leaves using Google Glass and a custom-developed software application together with a cost-effective leaf holder and multi-spectral illuminator device. Two images, taken using Google Glass, of a leaf placed in our portable illuminator device under red and white (i.e., broadband) light-emitting-diode (LED) illumination are uploaded to our servers for remote digital processing and chlorophyll quantification, with results returned to the user in less than 10 seconds. Intensity measurements extracted from the uploaded images are mapped against gold-standard colorimetric measurements made through a commercially available reader to generate calibration curves for plant leaf chlorophyll concentration. Using five plant species to calibrate our system, we demonstrate that our approach can accurately and rapidly estimate chlorophyll concentration of fifteen different plant species under both indoor and outdoor lighting conditions. This Google Glass based chlorophyll measurement platform can display the results in spatiotemporal and tabular forms and would be highly useful for monitoring of plant health in environmental and agriculture related applications, including e.g., urban plant monitoring, indirect measurements of the effects of climate change, and as an early indicator for water, soil, and air quality degradation. PMID:25669673

  17. Development and validation of a rapid turboflow LC-MS/MS method for the quantification of LSD and 2-oxo-3-hydroxy LSD in serum and urine samples of emergency toxicological cases.

    PubMed

    Dolder, Patrick C; Liechti, Matthias E; Rentsch, Katharina M

    2015-02-01

    Lysergic acid diethylamide (LSD) is a widely used recreational drug. The aim of the present study is to develop a quantitative turboflow LC-MS/MS method that can be used for rapid quantification of LSD and its main metabolite 2-oxo-3-hydroxy LSD (O-H-LSD) in serum and urine in emergency toxicological cases without time-consuming extraction steps. The method was developed on an ion-trap LC-MS/MS instrument coupled to a turbulent-flow extraction system. The validation data showed no significant matrix effects and no ion suppression has been observed in serum and urine. Mean intraday accuracy and precision for LSD were 101 and 6.84%, in urine samples and 97.40 and 5.89% in serum, respectively. For O-H-LSD, the respective values were 97.50 and 4.99% in urine and 107 and 4.70% in serum. Mean interday accuracy and precision for LSD were 100 and 8.26% in urine and 101 and 6.56% in serum, respectively. For O-H-LSD, the respective values were 101 and 8.11% in urine and 99.8 and 8.35% in serum, respectively. The lower limit of quantification for LSD was determined to be 0.1 ng/ml. LSD concentrations in serum were expected to be up to 8 ng/ml. 2-Oxo-3-hydroxy LSD concentrations in urine up to 250 ng/ml. The new method was accurate and precise in the range of expected serum and urine concentrations in patients with a suspected LSD intoxication. Until now, the method has been applied in five cases with suspected LSD intoxication where the intake of the drug has been verified four times with LSD concentrations in serum in the range of 1.80-14.70 ng/ml and once with a LSD concentration of 1.25 ng/ml in urine. In serum of two patients, the O-H-LSD concentration was determined to be 0.99 and 0.45 ng/ml. In the urine of a third patient, the O-H-LSD concentration was 9.70 ng/ml. PMID:25542574

  18. Colorimetric-spectral clustering: a tool for multispectral image compression

    NASA Astrophysics Data System (ADS)

    Ciprian, R.; Carbucicchio, M.

    2011-11-01

    In this work a new compression method for multispectral images has been proposed: the 'colorimetric-spectral clustering'. The basic idea arises from the well-known cluster analysis, a multivariate analysis which finds the natural links between objects grouping them into clusters. In the colorimetric-spectral clustering compression method, the objects are the spectral reflectance factors of the multispectral images that are grouped into clusters on the basis of their colour difference. In particular two spectra can belong to the same cluster only if their colour difference is lower than a threshold fixed before starting the compression procedure. The performance of the colorimetric-spectral clustering has been compared to the k-means cluster analysis, in which the Euclidean distance between spectra is considered, to the principal component analysis and to the LabPQR method. The colorimetric-spectral clustering is able to preserve both the spectral and the colorimetric information of a multispectral image, allowing this information to be reproduced for all pixels of the image.

  19. ``Red-to-blue'' colorimetric detection of cysteine via anti-etching of silver nanoprisms

    NASA Astrophysics Data System (ADS)

    Li, Yonglong; Li, Zihou; Gao, Yuexia; Gong, An; Zhang, Yujie; Hosmane, Narayan S.; Shen, Zheyu; Wu, Aiguo

    2014-08-01

    The reported strategies for cysteine (Cys) colorimetric detection based on noble metal nanomaterials include triggering aggregation, etching or fluorescence quenching of nanomaterials by Cys. In this study, we propose a new strategy for Cys colorimetric detection, i.e. anti-etching of silver nanoprisms (AgNPRs). In the absence of Cys, iodide ions (I-) could etch the corners and edges of AgNPRs and induce the morphology transition from nanoprism to nanodisk, which results in color change of the AgNPR dispersion from blue to red. In its presence, however, Cys can prevent the AgNPRs from I- attack. In that case, the color of the AgNPR dispersion containing I- and Cys remains blue. The mechanism is confirmed by using UV-vis spectra, TEM, DLS, Raman spectra and XPS spectra. According to the sensing effect of the Cys detection system, the concentration of I- incubated with AgNPRs, incubation time of AgNPRs and I-, and pH of AgNPR dispersions are optimized to 5.0 μM, 10 min, and pH 6.2, respectively. Under the optimized conditions, the proposed Cys detection system has excellent selectivity and high sensitivity. The limit of detection (LOD) of our Cys detection system is 25 nM by the naked eye, which is much better than the reported lowest LOD by eye-vision (100 nM), and 10 nM by UV-vis spectroscopy. The results of Cys detection in rabbit urine or plasma samples reinforce that our Cys detection system is applicable for rapid colorimetric detection of Cys in real body fluid samples.The reported strategies for cysteine (Cys) colorimetric detection based on noble metal nanomaterials include triggering aggregation, etching or fluorescence quenching of nanomaterials by Cys. In this study, we propose a new strategy for Cys colorimetric detection, i.e. anti-etching of silver nanoprisms (AgNPRs). In the absence of Cys, iodide ions (I-) could etch the corners and edges of AgNPRs and induce the morphology transition from nanoprism to nanodisk, which results in color change of the

  20. "Red-to-blue" colorimetric detection of cysteine via anti-etching of silver nanoprisms.

    PubMed

    Li, Yonglong; Li, Zihou; Gao, Yuexia; Gong, An; Zhang, Yujie; Hosmane, Narayan S; Shen, Zheyu; Wu, Aiguo

    2014-09-21

    The reported strategies for cysteine (Cys) colorimetric detection based on noble metal nanomaterials include triggering aggregation, etching or fluorescence quenching of nanomaterials by Cys. In this study, we propose a new strategy for Cys colorimetric detection, i.e. anti-etching of silver nanoprisms (AgNPRs). In the absence of Cys, iodide ions (I(-)) could etch the corners and edges of AgNPRs and induce the morphology transition from nanoprism to nanodisk, which results in color change of the AgNPR dispersion from blue to red. In its presence, however, Cys can prevent the AgNPRs from I(-) attack. In that case, the color of the AgNPR dispersion containing I(-) and Cys remains blue. The mechanism is confirmed by using UV-vis spectra, TEM, DLS, Raman spectra and XPS spectra. According to the sensing effect of the Cys detection system, the concentration of I(-) incubated with AgNPRs, incubation time of AgNPRs and I(-), and pH of AgNPR dispersions are optimized to 5.0 μM, 10 min, and pH 6.2, respectively. Under the optimized conditions, the proposed Cys detection system has excellent selectivity and high sensitivity. The limit of detection (LOD) of our Cys detection system is 25 nM by the naked eye, which is much better than the reported lowest LOD by eye-vision (100 nM), and 10 nM by UV-vis spectroscopy. The results of Cys detection in rabbit urine or plasma samples reinforce that our Cys detection system is applicable for rapid colorimetric detection of Cys in real body fluid samples. PMID:25083798

  1. Colorimetric assay for heterogeneous-catalyzed lipase activity: enzyme-regulated gold nanoparticle aggregation.

    PubMed

    Zhang, Wei; Tang, Yan; Liu, Jia; Jiang, Ling; Huang, Wei; Huo, Feng-Wei; Tian, Danbi

    2015-01-14

    Lipase is a neglected enzyme in the field of gold nanoparticle-based enzyme assays. This paper reports a novel colorimetric probe to rapidly visualize lipase activities by using Tween 20 functioned GNPs (Tween 20-GNPs) as a reporter. The present strategy hence could overcome the limitations caused by the heterogeneous interface in lipase assay. Catalytic hydrolytic cleavage of the ester bond in Tween 20-GNPs by lipase will trigger the rapid aggregation of GNPs at a high salt solution. The color change from red to purple could be used to sense the activity of lipase. The detection limit (3σ) is as low as 2.8 × 10-2 mg/mL. A preliminary enzyme activity screening was carried out for seven commercially purchased lipase samples. It also has been successfully applied to detecting lipase in fermentation broth of Bacillus subtilis without any pretreatment. PMID:25516269

  2. Smartphone based health accessory for colorimetric detection of biomarkers in sweat and saliva.

    PubMed

    Oncescu, Vlad; O'Dell, Dakota; Erickson, David

    2013-08-21

    The mobile health market is rapidly expanding and portable diagnostics tools offer an opportunity to decrease costs and increase the availability of healthcare. Here we present a smartphone based accessory and method for the rapid colorimetric detection of pH in sweat and saliva. Sweat pH can be correlated to sodium concentration and sweat rate in order to indicate to users the proper time to hydrate during physical exercise and avoid the risk of muscle cramps. Salivary pH below a critical threshold is correlated with enamel decalcification, an acidic breakdown of calcium in the teeth. We conduct a number of human trials with the device on a treadmill to demonstrate the ability to monitor changes in sweat pH due to exercise and electrolyte intake and predict optimal hydration. Additionally, we perform trials to measure salivary pH over time to monitor the effects of diet on oral health risks. PMID:23784453

  3. Accelerated colorimetric immunosensing using surface-modified porous monoliths and gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Chuag, Shao-Hsuan; Chen, Guan-Hua; Chou, Hsin-Hao; Shen, Shu-Wei; Chen, Chien-Fu

    2013-08-01

    A rapid and sensitive immunoassay platform integrating polymerized monoliths and gold nanoparticles (AuNPs) has been developed. The porous monoliths are photopolymerized in situ within a silica capillary and serve as solid support for high-mass transport and high-density capture antibody immobilization to create a shorter diffusion length for antibody-antigen interactions, resulting in a rapid assay and low reagent consumption. AuNPs are modified with detection antibodies and are utilized as signals for colorimetric immunoassays without the need for enzyme, substrate and sophisticated equipment for quantitative measurements. This platform has been verified by performing a human IgG sandwich immunoassay with a detection limit of 0.1 ng ml-1. In addition, a single assay can be completed in 1 h, which is more efficient than traditional immunoassays that require several hours to complete.

  4. Emergency First Responders' Experience with Colorimetric Detection Methods

    SciTech Connect

    Sandra L. Fox; Keith A. Daum; Carla J. Miller; Marnie M. Cortez

    2007-10-01

    Nationwide, first responders from state and federal support teams respond to hazardous materials incidents, industrial chemical spills, and potential weapons of mass destruction (WMD) attacks. Although first responders have sophisticated chemical, biological, radiological, and explosive detectors available for assessment of the incident scene, simple colorimetric detectors have a role in response actions. The large number of colorimetric chemical detection methods available on the market can make the selection of the proper methods difficult. Although each detector has unique aspects to provide qualitative or quantitative data about the unknown chemicals present, not all detectors provide consistent, accurate, and reliable results. Included here, in a consumer-report-style format, we provide “boots on the ground” information directly from first responders about how well colorimetric chemical detection methods meet their needs in the field and how they procure these methods.

  5. Lanthanide-labeled immunochromatographic strips for the rapid detection of Pantoea stewartii subsp. stewartii.

    PubMed

    Zhang, Fan; Zou, Mingqiang; Chen, Yan; Li, Jinfeng; Wang, Yanfei; Qi, Xiaohua; Xue, Qiang

    2014-01-15

    The lateral flow immunoassay is used in commercial pregnancy detection, and is an accepted point-of-care testing technique. The most widely used format for lateral flow immunochromatographic strips uses gold nanoparticles for colorimetric detection. However, this method often suffers from poor quantitative discrimination and low analytical sensitivity. To address these limitations, lanthanide chelate-loaded silica nanoparticles have been used as fluorescent labels. The fluorescent nanoparticles can easily bind to antibodies, with dextran as a linker. The strip reader described here was based on a sandwich immunoreaction performed on a strip, using lanthanide-labeled antibodies that served as signal vehicles for the fluorescent readout. The strip reader was used as a quantitative test system. In this work, Pantoea stewartii subsp. stewartii (Pss) was used as a model analyte to demonstrate the use of the strip reader. Under optimal conditions, the detection limit was determined as 10(3)cfu/mL. The quantification limit was calculated to be 10(4)cfu/mL. The detection limit for Pss was 100 times lower than those displayed by colloidal gold-labeled strips or ELISAs. No cross-reactions were observed with the other nine strains, indicating the good specificity of the Pss strip. This strip showed good stability in repeated tests. The tests using the fluorescence immunochromatographic strip were easy to perform, rapid, and sensitive. Methods using fluorescence strips and a strip reader have the potential to be a powerful tool for the quantification of bacteria. PMID:23928093

  6. Colorimetric blood-gas monitoring sensors

    NASA Astrophysics Data System (ADS)

    Proctor, Keith J.; Seifert, George P.

    1993-05-01

    Colorimetric fiber optic sensors have been developed for measuring the pH and pCO2 of blood. These sensors are fabricated using a single 125 micrometers diameter optical fiber. Located at the distal end of the fiber is a capsule that contains a pH sensitive dye. The pCO2 sensor is fabricated from a pH sensor with the addition of a salt, bicarbonate, and the encapsulation with an ion impermeable gas permeable membrane. The distal end of the capsule is terminated with a reflective surface. The reflective surface can either be a polished metallic surface or, in this case, a TiO2 impregnated epoxy. The disposable sensor mates with an optical connector that contains two optical fibers of the same size as the disposable sensor. The two fibers within the optical cable provide a light path for both the antegrade and retrograde optical signals. These fibers are terminated at either the LED source or the detector. A prototype sensor assembly that incorporates the measurement of three physiological parameters (pH, pCO2, and sO2) has been demonstrated to fit within a standard 20 gauge arterial catheter, typically used for radial artery blood pressure monitoring, without significant damping of the blood pressure waveform. The pH sensor has a range of 6.9 - 7.8 with a precision of 0.01 pH units and the pCO2 sensor has a range of 15 - 95 mm Hg with a precision of 3 mm Hg. The long term drift pH drift is less than 0.01 pH unit per 8 hours and the pCO2 drift is less than 1 mm Hg per 8 hours. Sensor performance in the canine has demonstrated that the pH sensor is accurate to within +/- 0.03 pH units and the pCO2 sensor is accurate to within +/- 3 mm Hg when compared to a typical blood gas analyzer.

  7. Automatic colorimetric calibration of human wounds

    PubMed Central

    2010-01-01

    Background Recently, digital photography in medicine is considered an acceptable tool in many clinical domains, e.g. wound care. Although ever higher resolutions are available, reproducibility is still poor and visual comparison of images remains difficult. This is even more the case for measurements performed on such images (colour, area, etc.). This problem is often neglected and images are freely compared and exchanged without further thought. Methods The first experiment checked whether camera settings or lighting conditions could negatively affect the quality of colorimetric calibration. Digital images plus a calibration chart were exposed to a variety of conditions. Precision and accuracy of colours after calibration were quantitatively assessed with a probability distribution for perceptual colour differences (dE_ab). The second experiment was designed to assess the impact of the automatic calibration procedure (i.e. chart detection) on real-world measurements. 40 Different images of real wounds were acquired and a region of interest was selected in each image. 3 Rotated versions of each image were automatically calibrated and colour differences were calculated. Results 1st Experiment: Colour differences between the measurements and real spectrophotometric measurements reveal median dE_ab values respectively 6.40 for the proper patches of calibrated normal images and 17.75 for uncalibrated images demonstrating an important improvement in accuracy after calibration. The reproducibility, visualized by the probability distribution of the dE_ab errors between 2 measurements of the patches of the images has a median of 3.43 dE* for all calibrated images, 23.26 dE_ab for all uncalibrated images. If we restrict ourselves to the proper patches of normal calibrated images the median is only 2.58 dE_ab! Wilcoxon sum-rank testing (p < 0.05) between uncalibrated normal images and calibrated normal images with proper squares were equal to 0 demonstrating a highly

  8. Colorimetric determination of Timolol concentration based on localized surface plasmon resonance of silver nanoparticles

    NASA Astrophysics Data System (ADS)

    Amirjani, Amirmostafa; Bagheri, Mozhgan; Heydari, Mojgan; Hesaraki, Saeed

    2016-09-01

    In this work, a rapid and simple colorimetric method based on the surface plasmon resonance of silver nanoparticles (AgNPs) was developed for the detection of the drug Timolol. The method used is based on the interaction of Timolol with the surface of the as‑synthesized AgNPs, which promotes aggregation of the nanoparticles. This aggregation exploits the surface plasmon resonance through the electric dipole–dipole interaction and coupling among the agglomerated particles, hence bringing forth distinctive changes in the spectra as well as the color of colloidal silver. UV‑vis spectrophotometery was used to monitor the changes of the localized surface plasmon resonance of AgNPs at wavelengths of 400 and 550 nm. The developed colorimetric sensor has a wide dynamic range of 1.0 × 10‑7 M–1.0 × 10‑3 M for detection of Timolol with a low detection limit of 1.2 × 10‑6 M. The proposed method was successfully applied for the determination of Timolol concentration in ophthalmic eye‑drop solution with a response time lower than 40 s.

  9. Sensitive colorimetric detection of Listeria monocytogenes based on isothermal gene amplification and unmodified gold nanoparticles.

    PubMed

    Fu, Zhongyu; Zhou, Xiaoming; Xing, Da

    2013-12-15

    Listeria monocytogenes (L. monocytogenes), one of most problematic food-borne bacteria, is mainly transmitted through the food chain and may cause listeriosis. Therefore, the development of rapid and sensitive L. monocytogenes detection technique has become an urgent task. In this study, we proposed a method using hyperbranching rolling circle amplification (HRCA) combined with gold nanoparticle (GNP) based colorimetric strategy to offer an isothermal, highly sensitive and specific assay for the detection of L. monocytogenes. First, a linear padlock probe targeting a specific sequence in the hly gene was designed and followed with a ligation by Taq DNA ligase. After ligation, further amplification by HRCA with a thiolated primer and an unlabeled primer is performed. The resulting thiolated HRCA products were then captured onto GNP surface and made GNP more salt-tolerant. Detection of the bacteria can be achieved by a facilitated GNP based colorimetric testing using naked eyes. Through this approach, as low as 100 aM synthetic hly gene targets and about 75 copies of L. monocytogenes can be detected. The specificity is evaluated by distinguishing target L. monocytogenes from other bacteria. The artificial contaminated food samples were also detected for its potential applications in real food detection. This method described here is ideal for bacteria detection due to its simplicity and high sensitivity. PMID:23948710

  10. Colorimetric determination of Timolol concentration based on localized surface plasmon resonance of silver nanoparticles.

    PubMed

    Amirjani, Amirmostafa; Bagheri, Mozhgan; Heydari, Mojgan; Hesaraki, Saeed

    2016-09-16

    In this work, a rapid and simple colorimetric method based on the surface plasmon resonance of silver nanoparticles (AgNPs) was developed for the detection of the drug Timolol. The method used is based on the interaction of Timolol with the surface of the as-synthesized AgNPs, which promotes aggregation of the nanoparticles. This aggregation exploits the surface plasmon resonance through the electric dipole-dipole interaction and coupling among the agglomerated particles, hence bringing forth distinctive changes in the spectra as well as the color of colloidal silver. UV-vis spectrophotometery was used to monitor the changes of the localized surface plasmon resonance of AgNPs at wavelengths of 400 and 550 nm. The developed colorimetric sensor has a wide dynamic range of 1.0 × 10(-7) M-1.0 × 10(-3) M for detection of Timolol with a low detection limit of 1.2 × 10(-6) M. The proposed method was successfully applied for the determination of Timolol concentration in ophthalmic eye-drop solution with a response time lower than 40 s. PMID:27504595

  11. A novel colorimetric biosensor for monitoring and detecting acute toxicity in water.

    PubMed

    Zhai, Junfeng; Yong, Daming; Li, Jing; Dong, Shaojun

    2013-01-21

    This work presents a new colorimetric microorganism biosensor for monitoring and detecting acute toxicity in water, where prussian blue (PB) is used as the colorimetric indicator and E. coli as the model bacterial. In this biosensor, the electron mediator, ferricyanide, accepts electrons from E. coli during respiration to produce ferrocyanide, which subsequently reacts with ferric ions to yield PB, a famous material with a blue color. Since toxicants can inhibit the respiratory activity of E. coli and then reduce the ferrocyanide and consequent PB production, toxicity can be easily detected by measuring the decrease in the production of PB induced by toxicants. Three important toxicants, 3,5-dichlorophenol (DCP), As(3+), Cr(6+) are tested and the detection limits are 3.2, 25, and 3.2 ppm, respectively. Moreover, we could identify the yellow green to dark green color change by naked eye even at concentrations as low as 12.5 ppm for both DCP and Cr(6+). Subsequently, the acute toxicities of groundwater and south lake water are successfully determined by this sensor. This biosensor is rapid, sensitive and cost-effective, and can thus be regarded as a promising biosensor for giving an early warning of acute water toxicity. PMID:23187797

  12. Functional gold nanoparticles coupled with microporous membranes: a flow controlled assay for colorimetric visualization of proteins.

    PubMed

    Chen, Yu-Yuan; Unnikrishnan, Binesh; Li, Yu-Jia; Huang, Chih-Ching

    2014-11-21

    We report a rapid and simple assay for colorimetric visualization of thrombin at nanomolar levels using functional gold nanoparticles (FAuNPs) coupled with microporous membranes. We used a 29-mer thiolated-thrombin-binding-aptamer (TBA29) to prepare TBA29 functionalized AuNPs (TBA29-AuNPs) for the selective detection of human thrombin. The sensing mechanism is based on the principle of TBA29-AuNPs flowing down through the nitrocellulose membrane (NCM) pores at different flow rates after binding to thrombin. Compared with free TBA29-AuNPs, when thrombin-TBA29-AuNPs were dropped on the NCM, the particles flowed down more easily through the NCM pores along with the buffer solution due to the increase in the gravity of particles. Therefore, color intensities of TBA29-AuNPs on the NCM depended on the concentration of thrombin; the color intensity was lighter when the concentration of thrombin was higher. Thrombin can be detected at the nanomolar level with the naked eye using this colorimetric probe. A protein G modified AuNP based probe (PG-AuNPs/NCM) was employed to detect human immunoglobulin G (hIgG) in plasma samples to demonstrate the practicality of our sensing system. Also, fibrinogen modified Au NPs were analyzed to demonstrate that this concept of detection could be extended to other proteins or systems, by functionalizing with suitable molecules. PMID:25267979

  13. Highly sensitive and specific colorimetric detection of cancer cells via dual-aptamer target binding strategy.

    PubMed

    Wang, Kun; Fan, Daoqing; Liu, Yaqing; Wang, Erkang

    2015-11-15

    Simple, rapid, sensitive and specific detection of cancer cells is of great importance for early and accurate cancer diagnostics and therapy. By coupling nanotechnology and dual-aptamer target binding strategies, we developed a colorimetric assay for visually detecting cancer cells with high sensitivity and specificity. The nanotechnology including high catalytic activity of PtAuNP and magnetic separation & concentration plays a vital role on the signal amplification and improvement of detection sensitivity. The color change caused by small amount of target cancer cells (10 cells/mL) can be clearly distinguished by naked eyes. The dual-aptamer target binding strategy guarantees the detection specificity that large amount of non-cancer cells and different cancer cells (10(4) cells/mL) cannot cause obvious color change. A detection limit as low as 10 cells/mL with detection linear range from 10 to 10(5) cells/mL was reached according to the experimental detections in phosphate buffer solution as well as serum sample. The developed enzyme-free and cost effective colorimetric assay is simple and no need of instrument while still provides excellent sensitivity, specificity and repeatability, having potential application on point-of-care cancer diagnosis. PMID:26042871

  14. Improving of enzyme immunoassay for detection and quantification of the target molecules using silver nanoparticles

    NASA Astrophysics Data System (ADS)

    Syrvatka, Vasyl J.; Slyvchuk, Yurij I.; Rozgoni, Ivan I.; Gevkan, Ivan I.; Overchuk, Marta O.

    2014-02-01

    Modern routine enzyme immunoassays for detection and quantification of biomolecules have several disadvantages such as high cost, insufficient sensitivity, complexity and long-term execution. The surface plasmon resonance of silver nanoparticles gives reasons of creating new in the basis of simple, highly sensitive and low cost colorimetric assays that can be applied to the detection of small molecules, DNA, proteins and pollutants. The main aim of the study was the improving of enzyme immunoassay for detection and quantification of the target molecules using silver nanoparticles. For this purpose we developed method for synthesis of silver nanoparticles with hyaluronic acid and studied possibility of use these nanoparticles in direct determination of target molecules concentration (in particular proteins) and for improving of enzyme immunoassay. As model we used conventional enzyme immunoassays for determination of progesterone and estradiol concentration. We obtained the possibility to produce silver nanoparticles with hyaluronan homogeneous in size between 10 and 12 nm, soluble and stable in water during long term of storage using modified procedure of silver nanoparticles synthesis. New method allows to obtain silver nanoparticles with strong optical properties at the higher concentrations - 60-90 μg/ml with the peak of absorbance at the wavelength 400 nm. Therefore surface plasmon resonance of silver nanoparticles with hyaluronan and ultraviolet-visible spectroscopy provide an opportunity for rapid determination of target molecules concentration (especial protein). We used silver nanoparticles as enzyme carriers and signal enhancers. Our preliminary data show that silver nanoparticles increased absorbance of samples that allows improving upper limit of determination of estradiol and progesterone concentration.

  15. A portable lab-on-a-chip system for gold-nanoparticle-based colorimetric detection of metal ions in water.

    PubMed

    Zhao, Chen; Zhong, Guowei; Kim, Da-Eun; Liu, Jinxia; Liu, Xinyu

    2014-09-01

    Heavy metal ions released into various water systems have a severe impact on the environment and human beings, and excess exposure to toxic metal ions through drinking water poses high risks to human health and causes life-threatening diseases. Thus, there is high demand for the development of a rapid, low-cost, and sensitive method for detection of metal ions in water. We present a portable analytical system for colorimetric detection of lead (Pb(2+)) and aluminum (Al(3+)) ions in water based on gold nanoparticle probes and lab-on-a-chip instrumentation. The colorimetric detection of metal ions is conducted via single-step assays with low limits of detection (LODs) and high selectivity. We design a custom-made microwell plate and a handheld colorimetric reader for implementing the assays and quantifying the signal readout. The calibration experiments demonstrate that this portable system provides LODs of 30 ppb for Pb(2+) and 89 ppb for Al(3+), both comparable to bench-top analytical spectrometers. It promises an effective platform for metal ion analysis in a more economical and convenient way, which is particularly useful for water quality monitoring in field and resource-poor settings. PMID:25332734

  16. Localized surface plasmon resonance of gold nanoparticles as colorimetric probes for determination of Isoniazid in pharmacological formulation

    NASA Astrophysics Data System (ADS)

    Zargar, Behrooz; Hatamie, Amir

    2013-04-01

    Isoniazid is an important antibiotic, which is widely used to treat tuberculosis. This study presents a colorimetric method for the determination of Isoniazid based on localized surface plasmon resonance (LSPR) property of gold nanoparticles. An LSPR band is produced by reducing gold ions in solution using Isoniazid as the reducing agent. Influences of the following relevant variables were examined and optimized in the experiment, formation time of gold nanoparticles, pH, buffer and stabilizer. These tests demonstrated that under optimum conditions the absorbance of Au nanoparticles at 530 nm related linearly to the concentration of Isoniazid in the range of 1.0-8.0 μg mL-1 with a detection limit of 0.98 μg mL-1. This colorimetric method has been successfully applied to the determine Isoniazid in tablets and spiked serum samples. The proposed colorimetric assay exhibits good reproducibility and accuracy, providing a simple and rapid method for analysis of Isoniazid.

  17. Resazurin Microtiter Assay Plate Testing of Mycobacterium tuberculosis Susceptibilities to Second-Line Drugs: Rapid, Simple, and Inexpensive Method

    PubMed Central

    Martin, Anandi; Camacho, Mirtha; Portaels, Françoise; Palomino, Juan Carlos

    2003-01-01

    The emergence of multidrug-resistant tuberculosis calls for new, rapid drug susceptibility tests. We have tested 150 Mycobacterium tuberculosis isolates against the second-line drugs ethionamide, kanamycin, capreomycin, ofloxacin, and para-aminosalicylic acid by the colorimetric resazurin microtiter assay and the proportion method. By visual reading, MICs were obtained after 8 days. A very good correlation between results by the colorimetric resazurin microtiter assay and the proportion method was obtained. The colorimetric resazurin microtiter assay is inexpensive, rapid, and simple to perform, and implementation of the assay is feasible for low-resource countries. PMID:14576129

  18. A colorimetric assay for alpha-hydroxynitrile lyase.

    PubMed

    Selmar, D; Carvalho, F J; Conn, E E

    1987-10-01

    A colorimetric assay for alpha-hydroxynitrile lyase which utilizes acetone cyanohydrin as a substrate is described. The assay is based on measurement of the HCN formed when the lyase catalyzes the dissociation of acetone cyanohydrin. The procedure was devised for use with the optically inactive acetone cyanohydrin but will be applicable to enzymes utilizing other cyanohydrins. PMID:3674409

  19. A modified colorimetric method for phytic acid analysis in soybean

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A quantitative, reproducible, and efficient phytic acid assay procedure is needed to screen breeding populations and support genetic studies in soybeans. The objective of this study was to modify the colorimetric Wade reagent method and compare the accuracy and applicability of this new method in de...

  20. Evaluation of a Colorimetric Personal Dosimeter for Nitrogen Oxide.

    ERIC Educational Resources Information Center

    Diamond, Philip

    A personal colorimetric dosimeter for nitrogen dioxide was developed. Tests were performed to determine the response of these strips to various concentrations of NO2. The dosimeter strips were satisfactory for approximate determinations of total exposure (concentration + time) of nitrogen dioxide. The total exposure was calculated in terms of time…

  1. Photonic crystal structures with tunable structure color as colorimetric sensors.

    PubMed

    Wang, Hui; Zhang, Ke-Qin

    2013-01-01

    Colorimetric sensing, which transduces environmental changes into visible color changes, provides a simple yet powerful detection mechanism that is well-suited to the development of low-cost and low-power sensors. A new approach in colorimetric sensing exploits the structural color of photonic crystals (PCs) to create environmentally-influenced color-changeable materials. PCs are composed of periodic dielectrics or metallo-dielectric nanostructures that affect the propagation of electromagnetic waves (EM) by defining the allowed and forbidden photonic bands. Simultaneously, an amazing variety of naturally occurring biological systems exhibit iridescent color due to the presence of PC structures throughout multi-dimensional space. In particular, some kinds of the structural colors in living organisms can be reversibly changed in reaction to external stimuli. Based on the lessons learned from natural photonic structures, some specific examples of PCs-based colorimetric sensors are presented in detail to demonstrate their unprecedented potential in practical applications, such as the detections of temperature, pH, ionic species, solvents, vapor, humidity, pressure and biomolecules. The combination of the nanofabrication technique, useful design methodologies inspired by biological systems and colorimetric sensing will lead to substantial developments in low-cost, miniaturized and widely deployable optical sensors. PMID:23539027

  2. Photonic Crystal Structures with Tunable Structure Color as Colorimetric Sensors

    PubMed Central

    Wang, Hui; Zhang, Ke-Qin

    2013-01-01

    Colorimetric sensing, which transduces environmental changes into visible color changes, provides a simple yet powerful detection mechanism that is well-suited to the development of low-cost and low-power sensors. A new approach in colorimetric sensing exploits the structural color of photonic crystals (PCs) to create environmentally-influenced color-changeable materials. PCs are composed of periodic dielectrics or metallo-dielectric nanostructures that affect the propagation of electromagnetic waves (EM) by defining the allowed and forbidden photonic bands. Simultaneously, an amazing variety of naturally occurring biological systems exhibit iridescent color due to the presence of PC structures throughout multi-dimensional space. In particular, some kinds of the structural colors in living organisms can be reversibly changed in reaction to external stimuli. Based on the lessons learned from natural photonic structures, some specific examples of PCs-based colorimetric sensors are presented in detail to demonstrate their unprecedented potential in practical applications, such as the detections of temperature, pH, ionic species, solvents, vapor, humidity, pressure and biomolecules. The combination of the nanofabrication technique, useful design methodologies inspired by biological systems and colorimetric sensing will lead to substantial developments in low-cost, miniaturized and widely deployable optical sensors. PMID:23539027

  3. A Colorimetric Process to Visualize Erythrocyte Exovesicles Aggregates

    ERIC Educational Resources Information Center

    Saldanha, Carlota; Santos, Nuno C.; Martins-Silva, J.

    2004-01-01

    A biochemistry laboratory class protocol is described in order to create an opportunity for students to apply by doing the theoretical concepts underlying biomolecules and vesicles properties, together with the principles of centrifugation and colorimetric methodologies. Through simple procedures the students will i) observe the segregation of the…

  4. Improvement on the polynomial modeling of digital camera colorimetric characterization

    NASA Astrophysics Data System (ADS)

    Huang, Xiaoqiao; Yu, Hongfei; Shi, Junsheng; Tai, Yonghang

    2014-11-01

    The digital camera has become a requisite for people's life, also essential in imaging applications, and it is important to get more accurate colors with digital camera. The colorimetric characterization of digital camera is the basis of image copy and color management process. One of the traditional methods for deriving a colorimetric mapping between camera RGB signals and the tristimulus values CIEXYZ is to use polynomial modeling with 3×11 polynomial transfer matrices. In this paper, an improved polynomial modeling is presented, in which the normalized luminance replaces the camera inherent RGB values in the traditional polynomial modeling. The improved modeling can be described by a two stage model. The first stage, relationship between the camera RGB values and normalized luminance with six gray patches in the X-rite ColorChecker 24-color card was described as "Gamma", camera RGB values were converted into normalized luminance using Gamma. The second stage, the traditional polynomial modeling was improved to the colorimetric mapping between normalized luminance and the CIEXYZ. Meanwhile, this method was used under daylight lighting environment, the users can not measure the CIEXYZ of the color target char using professional instruments, but they can accomplish the task of the colorimetric characterization of digital camera. The experimental results show that: (1) the proposed method for the colorimetric characterization of digital camera performs better than traditional polynomial modeling; (2) it's a feasible approach to handle the color characteristics using this method under daylight environment without professional instruments, the result can satisfy for request of simple application.

  5. Analysis of antifreeze protein activity using colorimetric gold nanosensors

    NASA Astrophysics Data System (ADS)

    Jing, Xu; Choi, Ho-seok; Park, Ji-In; Kim, Young-Pil

    2015-07-01

    High activity and long stability of antifreeze proteins (AFPs), also known as ice-binding proteins (IBPs), are necessary for exerting their physiological functions in biotechnology and cryomedicine. Here we report a simple analysis of antifreeze protein activity and stability based on self-assembly of gold nanoparticles (AuNPs) via freezing and thawing cycles. While the mercaptosuccinic acid-capped AuNP (MSA-AuNP) was easily self-assembled after a freezing/thawing cycle, due to the mechanical attack of ice crystal on the MSA-AuNP surface, the presence of AFP impeded the self-assembly of MSA-AuNP via the interaction of AFP with ice crystals via freezing and thawing cycles, which led to a strong color in the MSA-AuNP solution. As a result, the aggregation parameter (E520/E650) of MSA-AuNP showed the rapid detection of both activity and stability of AFPs. We suggest that our newly developed method is very suitable for measuring antifreeze activity and stability in a simple and rapid manner with reliable quantification.

  6. Simplified three-dimensional tissue clearing and incorporation of colorimetric phenotyping

    PubMed Central

    Sung, Kevin; Ding, Yichen; Ma, Jianguo; Chen, Harrison; Huang, Vincent; Cheng, Michelle; Yang, Cindy F.; Kim, Jocelyn T.; Eguchi, Daniel; Di Carlo, Dino; Hsiai, Tzung K.; Nakano, Atsushi; Kulkarni, Rajan P.

    2016-01-01

    Tissue clearing methods promise to provide exquisite three-dimensional imaging information; however, there is a need for simplified methods for lower resource settings and for non-fluorescence based phenotyping to enable light microscopic imaging modalities. Here we describe the simplified CLARITY method (SCM) for tissue clearing that preserves epitopes of interest. We imaged the resulting tissues using light sheet microscopy to generate rapid 3D reconstructions of entire tissues and organs. In addition, to enable clearing and 3D tissue imaging with light microscopy methods, we developed a colorimetric, non-fluorescent method for specifically labeling cleared tissues based on horseradish peroxidase conversion of diaminobenzidine to a colored insoluble product. The methods we describe here are portable and can be accomplished at low cost, and can allow light microscopic imaging of cleared tissues, thus enabling tissue clearing and imaging in a wide variety of settings. PMID:27498769

  7. Colorimetric detection of urine glucose based ZnFe2O4 magnetic nanoparticles.

    PubMed

    Su, Li; Feng, Jie; Zhou, Ximin; Ren, Cuiling; Li, Honghong; Chen, Xingguo

    2012-07-01

    In this paper, we discovered that ZnFe(2)O(4) magnetic nanoparticles (MNPs) possess intrinsic peroxidase-like activity. ZnFe(2)O(4) MNPs exhibit several advantages such as high catalytic efficiency, good stability, monodispersion, and rapid separation over other peroxidase nanomimetics and horseradish peroxidase (HRP). ZnFe(2)O(4) MNPs were used as a colorimetric biosensor for the detection of urine glucose. This method is simple, inexpensive, highly sensitive, and selective for glucose detection using glucose oxidase (GOx) and ZnFe(2)O(4) MNPs with a linear range from 1.25 × 10(-6) to 1.875 × 10(-5) mol L(-1) with a detection limit of 3.0 × 10(-7) mol L(-1). The color change observable by the naked eyes based on the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) is the principle for the sensing of urine glucose level. PMID:22702236

  8. Preparation of a novel colorimetric luminescence sensor strip for the detection of indole-3-acetic acid.

    PubMed

    Liu, Yan; Dong, Haitao; Zhang, Wenzhu; Ye, Zhiqiang; Wang, Guilan; Yuan, Jingli

    2010-06-15

    A novel colorimetric luminescence sensor strip for the detection of indole-3-acetic acid (IAA) has been fabricated by using green emissive quantum dots of cadmium telluride (CdTe QDs) as a background layer and a red emissive europium chelate, [4'-(9-anthryl)-2,2':6',2''-terpyridine-6,6''-diyl]bis(methylenenitrilo) tetrakis(acetate)-Eu(3+) (ATTA-Eu(3+)), as a specific sensing layer coated on the surface of glass slide, respectively. The luminescence response of the sensor strip is given by the dramatic changes in emission colors from green to red at different IAA concentrations. This approach provides a simple, rapid, sensitive and accurate method for the detection of IAA without using any special scientific instruments. PMID:20353890

  9. Colorimetric As (V) detection based on S-layer functionalized gold nanoparticles.

    PubMed

    Lakatos, Mathias; Matys, Sabine; Raff, Johannes; Pompe, Wolfgang

    2015-11-01

    Herein, we present simple and rapid colorimetric and UV/VIS spectroscopic methods for detecting anionic arsenic (V) complexes in aqueous media. The methods exploit the aggregation of S-layer-functionalized spherical gold nanoparticles of sizes between 20 and 50 nm in the presence of arsenic species. The gold nanoparticles were functionalized with oligomers of the S-layer protein of Lysinibacillus sphaericus JG-A12. The aggregation of the nanoparticles results in a color change from burgundy-red for widely dispersed nanoparticles to blue for aggregated nanoparticles. A detailed signal analysis was achieved by measuring the shift of the particle plasmon resonance signal with UV/VIS spectroscopy. To further improve signal sensitivity, the influence of larger nanoparticles was tested. In the case of 50 nm gold nanoparticles, a concentration of the anionic arsenic (V) complex lower than 24 ppb was detectable. PMID:26452816

  10. Simplified three-dimensional tissue clearing and incorporation of colorimetric phenotyping.

    PubMed

    Sung, Kevin; Ding, Yichen; Ma, Jianguo; Chen, Harrison; Huang, Vincent; Cheng, Michelle; Yang, Cindy F; Kim, Jocelyn T; Eguchi, Daniel; Di Carlo, Dino; Hsiai, Tzung K; Nakano, Atsushi; Kulkarni, Rajan P

    2016-01-01

    Tissue clearing methods promise to provide exquisite three-dimensional imaging information; however, there is a need for simplified methods for lower resource settings and for non-fluorescence based phenotyping to enable light microscopic imaging modalities. Here we describe the simplified CLARITY method (SCM) for tissue clearing that preserves epitopes of interest. We imaged the resulting tissues using light sheet microscopy to generate rapid 3D reconstructions of entire tissues and organs. In addition, to enable clearing and 3D tissue imaging with light microscopy methods, we developed a colorimetric, non-fluorescent method for specifically labeling cleared tissues based on horseradish peroxidase conversion of diaminobenzidine to a colored insoluble product. The methods we describe here are portable and can be accomplished at low cost, and can allow light microscopic imaging of cleared tissues, thus enabling tissue clearing and imaging in a wide variety of settings. PMID:27498769

  11. Colorimetric Detection of Small Molecules in Complex Matrixes via Target-Mediated Growth of Aptamer-Functionalized Gold Nanoparticles.

    PubMed

    Soh, Jun Hui; Lin, Yiyang; Rana, Subinoy; Ying, Jackie Y; Stevens, Molly M

    2015-08-01

    A versatile and sensitive colorimetric assay that allows the rapid detection of small-molecule targets using the naked eye is demonstrated. The working principle of the assay integrates aptamer-target recognition and the aptamer-controlled growth of gold nanoparticles (Au NPs). Aptamer-target interactions modulate the amount of aptamer strands adsorbed on the surface of aptamer-functionalized Au NPs via desorption of the aptamer strands when target molecules bind with the aptamer. Depending on the resulting aptamer coverage, Au NPs grow into morphologically varied nanostructures, which give rise to different colored solutions. Au NPs with low aptamer coverage grow into spherical NPs, which produce red-colored solutions, whereas Au NPs with high aptamer coverage grow into branched NPs, which produce blue-colored solutions. We achieved visible colorimetric response and nanomolar detection limits for the detection of ochratoxin A (1 nM) in red wine samples, as well as cocaine (1 nM) and 17β-estradiol (0.2 nM) in spiked synthetic urine and saliva, respectively. The detection limits were well within clinically and physiologically relevant ranges, and below the maximum food safety limits. The assay is highly sensitive, specific, and able to detect an array of analytes rapidly without requiring sophisticated equipment, making it relevant for many applications, such as high-throughput drug and clinical screening, food sampling, and diagnostics. Furthermore, the assay is easily adapted as a chip-based platform for rapid and portable target detection. PMID:26197040

  12. Phenylboronic acid modified silver nanoparticles for colorimetric dynamic analysis of glucose.

    PubMed

    Cao, Ke; Jiang, Xiaomei; Yan, Suting; Zhang, Laiying; Wu, Weitai

    2014-02-15

    The development of advanced nanostructures that allow dynamic quantification of glucose level can contribute to tight glucose control in diabetes management and other medical/biological fields. In this paper, we demonstrated that the assemblies of the 5-amino-2-fluorophenylboronic acid modified silver nanoparticles (FPBA-AgNPs) can be employed for highly modulating, sensitive, and selective colorimetric sensing of glucose over a physiologically important concentration range of 0-20mM at a physiological pH of 7.4. The glucose-modulated assembly of the FPBA-AgNPs occurred by the regulable formation of interparticle linkages via the bridged binding of 1,2-cis-diols and 5,6-cis-diols (for furanose form; or 4,6-cis-diols for pyranose form), respectively, of a glucose molecule to two FPBA-AgNPs. The detection limit was 89.0 μM. The mean error of glucose detection in a macro-bio-system, blood serum of adult, was smaller than 10%. Furthermore, we show that the glucose level variations associated with a model biological reaction process can be monitored by using the FPBA-AgNPs, whilst with the reaction mechanism remaining nearly unchanged. PMID:24055932

  13. Colorimetric determination of melamine in milk using unmodified silver nanoparticles

    NASA Astrophysics Data System (ADS)

    Kumar, Naveen; Kumar, Harish; Mann, Bimlesh; Seth, Raman

    2016-03-01

    Melamine is nitrogen rich chemical compound used as an adulterant in dairy products by unscrupulous people to increase the apparent protein content. This incident prompted the researchers to develop simple methods for easy detection of melamine in food samples. In the present paper, we report a simple and sensitive colorimetric method for detection of melamine in milk based on silver nanoparticles. This method relies upon the principle that melamine causes the aggregation of silver nanoparticles, resulting in abrupt color change from yellow to red under optimized conditions. The concentration of melamine in adulterated sample can be quantitated by monitoring the absorption spectra of silver nanoparticles using ultraviolet-visible (UV-Vis) spectrometer. The present colorimetric method which utilizes silver nanoparticles of 35 nm can reliably detect melamine down to a concentration of 0.04 mg l- 1.

  14. Colorimetric determination of melamine in milk using unmodified silver nanoparticles.

    PubMed

    Kumar, Naveen; Kumar, Harish; Mann, Bimlesh; Seth, Raman

    2016-03-01

    Melamine is nitrogen rich chemical compound used as an adulterant in dairy products by unscrupulous people to increase the apparent protein content. This incident prompted the researchers to develop simple methods for easy detection of melamine in food samples. In the present paper, we report a simple and sensitive colorimetric method for detection of melamine in milk based on silver nanoparticles. This method relies upon the principle that melamine causes the aggregation of silver nanoparticles, resulting in abrupt color change from yellow to red under optimized conditions. The concentration of melamine in adulterated sample can be quantitated by monitoring the absorption spectra of silver nanoparticles using ultraviolet-visible (UV-Vis) spectrometer. The present colorimetric method which utilizes silver nanoparticles of 35 nm can reliably detect melamine down to a concentration of 0.04 mg l(-1). PMID:26654965

  15. Azomethine H colorimetric method for determining dissolved boron in water

    USGS Publications Warehouse

    Spencer, R.R.; Erdmann, D.E.

    1979-01-01

    An automated colorimetric method for determining dissolved boron in water is described. The boron is complexed with azomethine H, which is readily available as the condensation product of H acid (8-amino-1-naphthol-3,6-disulfonic acid) and salicylaldehyde. The absorbance of the yellow complex formed is then measured colorimetrically at 410 nm. Interference effects from other dissolved species are minimized by the addition of diethylenetriaminepentaacetic acid (DTPA); however, iron, zinc, and bicarbonate interfere at concentrations above 400 ??g/L, 2000 ??g/L, and 200 mg/L, respectively. The bicarbonate interference can be eliminated by careful acidification of the sample with concentrated HCl to a pH between 5 and 6. Thirty samples per hour can be routinely analyzed over the range of from 10 to 400 ??g/L, boron.

  16. A selective colorimetric and ratiometric fluorescent probe for hydrogen sulfide.

    PubMed

    Wu, Ming-Yu; Li, Kun; Hou, Ji-Ting; Huang, Zheng; Yu, Xiao-Qi

    2012-10-01

    A reaction-based colorimetric and ratiometric fluorescent probe based on an ICT-strategy for selective detection of H(2)S that exploited the H(2)S-mediated reduction of nitrocompound to amines was explored. And it displayed high selectivity for H(2)S over other relevant reactive sulfur, oxygen, nitrogen species and other anions with more than 120 nm blue shift and the change of emission intensity ratio inducted by H(2)S was over 4750. PMID:22965805

  17. Surface plasmon resonance based selective and sensitive colorimetric determination of azithromycin using unmodified silver nanoparticles in pharmaceuticals and human plasma.

    PubMed

    Chavada, Vijay D; Bhatt, Nejal M; Sanyal, Mallika; Shrivastav, Pranav S

    2017-01-01

    In this article we report a novel method for colorimetric sensing and selective determination of a non-chromophoric drug-azithromycin, which lacks native absorbance in the UV-Visible region using unmodified silver nanoparticles (AgNPs). The citrate-capped AgNps dispersed in water afforded a bright yellow colour owing to the electrostatic repulsion between the particles due to the presence of negatively charged surface and showed surface plasmon resonance (SPR) band at 394nm. Addition of positively charged azithromycin at a concentration as low as 0.2μM induced rapid aggregation of AgNPs by neutralizing the negative charge on the particle surface. This phenomenon resulted in the colour change from bright yellow to purple which could be easily observed by the naked eye. This provided a simple platform for rapid determination of azithromycin based on colorimetric measurements. The factors affecting the colorimetric response like pH, volume of AgNPs suspension and incubation time were suitably optimized. The validated method was found to work efficiently in the established concentration range of 0.2-100.0μM using two different calibration models. The selectivity of the method was also evaluated by analysis of nanoparticles-aggregation response upon addition of several anions, cations and some commonly prescribed antibiotics. The method was successfully applied for the analysis of azithromycin in pharmaceuticals and spiked human plasma samples with good accuracy and precision. The simplicity, efficiency and cost-effectiveness of the method hold tremendous potential for the analysis of such non-chromophoric pharmaceuticals. PMID:27419643

  18. Simple Colorimetric Determination of the Manganese Content in Photosynthetic Membranes

    SciTech Connect

    Semin, B. K.; Seibert, M.

    2009-01-01

    The functional Mn content of intact photosystem II membrane fragments was measured as 4.06 {+-} 0.13 Mn/reaction center when determined using a simple, sensitive colorimetric assay that will also work with thylakoids and core complexes. This procedure requires minimal sample material, does not need expensive assay equipment, requires four simple steps, and only takes 20-30 min to perform. These include (a) removal of the adventitious Mn ions by CaCl{sub 2} treatment of the membranes, (b) extraction of the Mn from the O{sub 2}-evolving complex with hydrochloric acid, (c) purification of the extract by centrifugation followed by filtration of the supernatant through an Acrodisc syringe filter (0.2 {micro}m nylon membrane), and (d) colorimetric determination of Mn in the extract using the reaction of the chromogenic agent, 3,3',5,5'-tetramethylbenzidine, with previously oxidized Mn(II) cations carried out at high pH. The colorimetric assay itself has been used previously by Serrat (Mikrochim Acta 129:77-80, 1998) for assaying Mn concentrations in sea water and drinking water.

  19. Discrimination of Dental Caries Using Colorimetric Characteristics of Fluorescence Spectrum.

    PubMed

    Chen, Qingguang; Zhu, Haihua; Xu, Ying; Lin, Bin; Chen, Hui

    2015-01-01

    The feasibility of colorimetric parameters for the discrimination of the stages of dental caries based on a light-induced autofluorescence spectrum at a 405-nm excitation wavelength was investigated. The fluorescence spectra of 4 groups of tooth samples (10 sound, 10 early-stage decay, 14 established decay, and 10 severe decay), which were classified by the International Caries Detection and Assessment System, were experimentally measured in vitro. The carious lesion samples had an additional fluorescence peak at around 627 nm. The mathematical relation of the fluorescence spectrum and human color perception was established and computed. With increasing severity, the fluorescence color changed from green to yellow according to the colorimetric parameters of the CIE 1931 (x, y) chromaticity coordinates and dominant wavelengths. The results from a one-way ANOVA of the dominant wavelength showed a statistically significant difference among the 4 classified groups. The colorimetric parameters of the light-induced fluorescence spectrum can potentially be applied to evaluate the various carious levels. PMID:26112288

  20. Functional self-assembling bolaamphiphilic polydiacetylenes as colorimetric sensor scaffolds

    SciTech Connect

    Song, Jie; Cisar, Justin S.; Bertozzi, Carolyn R.

    2004-05-28

    Conjugated polymers capable of responding to external stimuli by changes in optical, electrical or electrochemical properties can be used for the construction of direct sensing devices. Polydiacetylene-based systems are attractive for sensing applications due to their colorimetric response to changes in the local environment. Here we present the design, preparation and characterization of self-assembling functional bolaamphiphilic polydiacetylenes (BPDAs) inspired by Nature's strategy for membrane stabilization. We show that by placing polar headgroups on both ends of the diacetylene lipids in a transmembranic fashion, and altering the chemical nature of the polar surface residues, the conjugated polymers can be engineered to display a range of radiation-, thermal- and pH-induced colorimetric responses. We observed dramatic nanoscopic morphological transformations accompanying charge-induced chromatic transitions, suggesting that both side chain disordering and main chain rearrangement play important roles in altering the effective conjugation lengths of the poly(ene-yne). These results establish the foundation for further development of BPDA-based colorimetric sensors.

  1. Colorimetric TMPRSS2-ERG Gene Fusion Detection in Prostate Cancer Urinary Samples via Recombinase Polymerase Amplification

    PubMed Central

    Koo, Kevin M.; Wee, Eugene J.H.; Trau, Matt

    2016-01-01

    TMPRSS2 (Exon 1)-ERG (Exon 4) is the most frequent gene fusion event in prostate cancer (PC), and is highly PC-specific unlike the current serum prostate specific antigen (PSA) biomarker. However, TMPRSS2-ERG levels are currently measured with quantitative reverse-transcription PCR (RT-qPCR) which is time-consuming and requires costly equipment, thus limiting its use in clinical diagnostics. Herein, we report a novel rapid, cost-efficient and minimal-equipment assay named “FusBLU” for detecting TMPRSS2-ERG gene fusions from urine. TMPRSS2-ERG mRNA was amplified by isothermal reverse transcription-recombinase polymerase amplification (RT-RPA), magnetically-isolated, and detected through horseradish peroxidase (HRP)-catalyzed colorimetric reaction. FusBLU was specific for TMPRSS2-ERG mRNA with a low visual detection limit of 105 copies. We also demonstrated assay readout versatility on 3 potentially useful platforms. The colorimetric readout was detectable by naked eye for a quick yes/no evaluation of gene fusion presence. On the other hand, a more quantitative TMPRSS2-ERG detection was achievable by absorbance/electrochemical measurements. FusBLU was successfully applied to 12 urinary samples and results were validated by gold-standard RT-qPCR. We also showed that sediment RNA was likely the main source of TMPRSS2-ERG mRNA in urinary samples. We believe that our assay is a potential clinical screening tool for PC and could also have wide applications for other disease-related fusion genes. PMID:27375789

  2. Colorimetric TMPRSS2-ERG Gene Fusion Detection in Prostate Cancer Urinary Samples via Recombinase Polymerase Amplification.

    PubMed

    Koo, Kevin M; Wee, Eugene J H; Trau, Matt

    2016-01-01

    TMPRSS2 (Exon 1)-ERG (Exon 4) is the most frequent gene fusion event in prostate cancer (PC), and is highly PC-specific unlike the current serum prostate specific antigen (PSA) biomarker. However, TMPRSS2-ERG levels are currently measured with quantitative reverse-transcription PCR (RT-qPCR) which is time-consuming and requires costly equipment, thus limiting its use in clinical diagnostics. Herein, we report a novel rapid, cost-efficient and minimal-equipment assay named "FusBLU" for detecting TMPRSS2-ERG gene fusions from urine. TMPRSS2-ERG mRNA was amplified by isothermal reverse transcription-recombinase polymerase amplification (RT-RPA), magnetically-isolated, and detected through horseradish peroxidase (HRP)-catalyzed colorimetric reaction. FusBLU was specific for TMPRSS2-ERG mRNA with a low visual detection limit of 10(5) copies. We also demonstrated assay readout versatility on 3 potentially useful platforms. The colorimetric readout was detectable by naked eye for a quick yes/no evaluation of gene fusion presence. On the other hand, a more quantitative TMPRSS2-ERG detection was achievable by absorbance/electrochemical measurements. FusBLU was successfully applied to 12 urinary samples and results were validated by gold-standard RT-qPCR. We also showed that sediment RNA was likely the main source of TMPRSS2-ERG mRNA in urinary samples. We believe that our assay is a potential clinical screening tool for PC and could also have wide applications for other disease-related fusion genes. PMID:27375789

  3. A colorimetric detection of acrylamide in potato chips based on nucleophile-initiated thiol-ene Michael addition.

    PubMed

    Hu, Qinqin; Fu, Yingchun; Xu, Xiahong; Qiao, Zhaohui; Wang, Ronghui; Zhang, Ying; Li, Yanbin

    2016-02-01

    Acrylamide (AA), a neurotoxin and a potential carcinogen, has been found in various thermally processed foods such as potato chips, biscuits, and coffee. Simple, cost-effective, and sensitive methods for the rapid detection of AA are needed to ensure food safety. Herein, a novel colorimetric method was proposed for the visual detection of AA based on a nucleophile-initiated thiol-ene Michael addition reaction. Gold nanoparticles (AuNPs) were aggregated by glutathione (GSH) because of a ligand-replacement, accompanied by a color change from red to purple. In the presence of AA, after the thiol-ene Michael addition reaction between GSH and AA with the catalysis of a nucleophile, the sulfhydryl group of GSH was consumed by AA, which hindered the subsequent ligand-replacement and the aggregation of AuNPs. Therefore, the concentration of AA could be determined by the visible color change caused by dispersion/aggregation of AuNPs. This new method showed high sensitivity with a linear range from 0.1 μmol L(-1) to 80 μmol L(-1) and a detection limit of 28.6 nmol L(-1), and especially revealed better selectivity than the fluorescence sensing method reported previously. Moreover, this new method was used to detect AA in potato chips with a satisfactory result in comparison with the standard methods based on chromatography, which indicated that the colorimetric method can be expanded for the rapid detection of AA in thermally processed foods. PMID:26699696

  4. Development of a VHH-Based Erythropoietin Quantification Assay.

    PubMed

    Kol, Stefan; Kallehauge, Thomas Beuchert; Adema, Simon; Hermans, Pim

    2015-08-01

    Erythropoietin (EPO) quantification during cell line selection and bioreactor cultivation has traditionally been performed with ELISA or HPLC. As these techniques suffer from several drawbacks, we developed a novel EPO quantification assay. A camelid single-domain antibody fragment directed against human EPO was evaluated as a capturing antibody in a label-free biolayer interferometry-based quantification assay. Human recombinant EPO can be specifically detected in Chinese hamster ovary cell supernatants in a sensitive and pH-dependent manner. This method enables rapid and robust quantification of EPO in a high-throughput setting. PMID:25764454

  5. A Sensitive, Colorimetric, High-Throughput Loop-Mediated Isothermal Amplification Assay for the Detection of Plasmodium knowlesi.

    PubMed

    Britton, Sumudu; Cheng, Qin; Grigg, Matthew J; William, Timothy; Anstey, Nicholas M; McCarthy, James S

    2016-07-01

    The simian parasite Plasmodium knowlesi is now the commonest cause of malaria in Malaysia and can rapidly cause severe and fatal malaria. However, microscopic misdiagnosis of Plasmodium species is common, rapid antigen detection tests remain insufficiently sensitive and confirmation of P. knowlesi requires polymerase chain reaction (PCR). Thus available point-of-care diagnostic tests are inadequate. This study reports the development of a simple, sensitive, colorimetric, high-throughput loop-mediated isothermal amplification assay (HtLAMP) diagnostic test using novel primers for the detection of P. knowlesi. This assay is able to detect 0.2 parasites/μL, and compared with PCR has a sensitivity of 96% for the detection of P. knowlesi, making it a potentially field-applicable point-of-care diagnostic tool. PMID:27162264

  6. Cost Effective Paper-Based Colorimetric Microfluidic Devices and Mobile Phone Camera Readers for the Classroom

    ERIC Educational Resources Information Center

    Koesdjojo, Myra T.; Pengpumkiat, Sumate; Wu, Yuanyuan; Boonloed, Anukul; Huynh, Daniel; Remcho, Thomas P.; Remcho, Vincent T.

    2015-01-01

    We have developed a simple and direct method to fabricate paper-based microfluidic devices that can be used for a wide range of colorimetric assay applications. With these devices, assays can be performed within minutes to allow for quantitative colorimetric analysis by use of a widely accessible iPhone camera and an RGB color reader application…

  7. Colorimetric elastase sensor with peptide conjugated cellulose nanocrystals is interfaced to dialysis membranes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clinical detection of human neutrophil elastase (HNE) as point of care biomarker or in situ colorimetric adjuvant to chronic wound dressings presents potential advantages in the management of chronic wounds. A colorimetric approach to the detection of HNE using cotton cellulose nanocrystals (CCN) i...

  8. Colorimetric Detection and Identification of Natural and Artificial Sweeteners

    PubMed Central

    Musto, Christopher J.; Lim, Sung H.; Suslick, Kenneth S.

    2009-01-01

    A disposable, low-cost colorimetric sensor array has been created by pin-printing onto a hydrophilic membrane 16 chemically responsive nanoporous pigments made from indicators immobilized in an organically modified silane (ormosil). The array has been used to detect and identify 14 different natural and artificial sweeteners at millimolar concentrations as well as commonly used individual serving sweetener packets. The array has shown excellent reproducibility and long shelf-life and has been optimized to work in the biological pH regime. PMID:20337402

  9. A non-aggregation colorimetric assay for thrombin based on catalytic properties of silver nanoparticles.

    PubMed

    Li, Jie; Li, Wei; Qiang, Weibing; Wang, Xi; Li, Hui; Xu, Danke

    2014-01-01

    In this paper, we developed a simple and rapid colorimetric assay for protein detection based on the reduction of dye molecules catalyzed by silver nanoparticles (AgNPs). Aptamer-modified magnetic particles and aptamer-functionalized AgNPs were employed as capture and detection probes, respectively. Introduction of thrombin as target protein could form a sandwich-type complex involving catalytically active AgNPs, whose catalytic activity was monitored on the catalytic reduction of rhodamine B (RhB) by sodium borohydride (NaBH4). The amount of immobilized AgNPs on the complex increased along with the increase of the thrombin concentration, thus the detection of thrombin was achieved via recording the decrease in absorbance corresponding to RhB. This method has adopted several advantages from the key factors involved, i.e., the sandwich binding of affinity aptamers contributed to the increased specificity; magnetic particles could result in rapid capture and separation processes; the conjugation of AgNPs would lead to a clear visual detection. It allows for the detection limit of thrombin down to picomolar level by the naked eye, with remarkable selectivity over other proteins. Moreover, it is possible to apply this method to the other targets with two binding sites as well. PMID:24356228

  10. Biofunctionalized gold nanoparticles for colorimetric sensing of botulinum neurotoxin A light chain.

    PubMed

    Liu, Xiaohu; Wang, Yi; Chen, Peng; Wang, Yusong; Zhang, Jinling; Aili, Daniel; Liedberg, Bo

    2014-03-01

    Botulinum neurotoxin is considered as one of the most toxic food-borne substances and is a potential bioweapon accessible to terrorists. The development of an accurate, convenient, and rapid assay for botulinum neurotoxins is therefore highly desirable for addressing biosafety concerns. Herein, novel biotinylated peptide substrates designed to mimic synaptosomal-associated protein 25 (SNAP-25) are utilized in gold nanoparticle-based assays for colorimetric detection of botulinum neurotoxin serotype A light chain (BoLcA). In these proteolytic assays, biotinylated peptides serve as triggers for the aggregation of gold nanoparticles, while the cleavage of these peptides by BoLcA prevents nanoparticle aggregation. Two different assay strategies are described, demonstrating limits of detection ranging from 5 to 0.1 nM of BoLcA with an overall assay time of 4 h. These hybrid enzyme-responsive nanomaterials provide rapid and sensitive detection for one of the most toxic substances known to man. PMID:24484451

  11. A new diatom growth inhibition assay using the XTT colorimetric method.

    PubMed

    Jiang, Weina; Akagi, Takuya; Suzuki, Hidekazu; Takimoto, Ayaka; Nagai, Hiroshi

    2016-01-01

    Marine biofouling, which leads to significant operational stress and economic damage on marine infrastructures, is a major problem in marine related industries. Currently, the most common way to avoid marine biofouling involves the use of biocidal products in surface coatings. However, the need for environmentally friendly antibiofouling compounds has increased rapidly with the recent global prohibition of harmful antifoulants, such as tributyltin (TBT). In particular, periphytic diatoms have been shown to contribute significantly to biofilms, which play an important role in biofouling. Therefore, inhibiting the proliferation of fouling diatoms is a very important step in the prevention of marine biofouling. In this study, we developed a new, rapid, accurate, and convenient growth inhibition assay using the XTT colorimetric method to prevent the growth of the fouling periphytic diatom, Nitzschia amabilis Hidek. Suzuki (replaced synonym, Nitzschia laevis Hustedt). The feasibility of this method was verified by determining the growth inhibition activities of two standard photosynthetic inhibitors, DCMU and CuSO4. However, neither inhibitor had any cytotoxic activities at the range of concentrations tested. Moreover, this method was applied by screening and purification of herbicidic but non-cytotoxic compounds from cyanobacteria extracts. Our results demonstrate the utility of this newly established growth inhibition assay for the identification of marine anti-biofouling compounds. PMID:26945522

  12. Evaluation of a colorimetric reagent strip assay for urine specific gravity.

    PubMed

    Kirschbaum, B B

    1983-06-01

    N-Multistix SG provides a convenient colorimetric assay for the determination of the specific gravity (sp. gr.) of freshly voided urine. When compared with results obtained by standard hydrometry, the colorimetric assay sp. gr. was observed to decrease by as much as 0.010 units as urine pH increased from 5 to 7. Moderate levels of proteinuria that did not alter hydrometer readings effectively raised the colorimetric sp. gr. by 0.005-0.010 units. The colorimetric assay was almost completely insensitive to clinically encountered concentrations of glucose and urea but responded appropriately to monovalent salts. The magnitude of these observed discrepancies places serious limitations on the value of the colorimetric sp. gr. measurement. PMID:6846262

  13. A lab-in-a-briefcase for rapid prostate specific antigen (PSA) screening from whole blood.

    PubMed

    Barbosa, Ana I; Castanheira, Ana P; Edwards, Alexander D; Reis, Nuno M

    2014-08-21

    We present a new concept for rapid and fully portable prostate specific antigen (PSA) measurements, termed "lab-in-a-briefcase", which integrates an affordable microfluidic ELISA platform utilising a melt-extruded fluoropolymer microcapillary film (MCF) containing an array of 10 200 μm internal diameter capillaries, a disposable multi-syringe aspirator (MSA), a sample tray pre-loaded with all of the required immunoassay reagents, and a portable film scanner for colorimetric signal digital quantification. Each MSA can perform 10 replicate microfluidic immunoassays on 8 samples, allowing 80 measurements to be made in less than 15 minutes based on semi-automated operation, without the need of additional fluid handling equipment. The assay was optimised for the measurement of a clinically relevant range of PSA of 0.9 to 60.0 ng ml(-1) in 15 minutes with CVs on the order of 5% based on intra-assay variability when read using a consumer flatbed film scanner. The PSA assay performance in the MSA remained robust in undiluted or 1 : 2 diluted human serum or whole blood, and the matrix effect could simply be overcome by extending sample incubation times. The PSA "lab-in-a-briefcase" is particularly suited to a low-resource health setting, where diagnostic labs and automated immunoassay systems are not accessible, by allowing PSA measurement outside the laboratory using affordable equipment. PMID:24989886

  14. HPAEC-PAD quantification of Haemophilus influenzae type b polysaccharide in upstream and downstream samples.

    PubMed

    van der Put, Robert M F; de Haan, Alex; van den IJssel, Jan G M; Hamidi, Ahd; Beurret, Michel

    2015-11-27

    Due to the rapidly increasing introduction of Haemophilus influenzae type b (Hib) and other conjugate vaccines worldwide during the last decade, reliable and robust analytical methods are needed for the quantitative monitoring of intermediate samples generated during fermentation (upstream processing, USP) and purification (downstream processing, DSP) of polysaccharide vaccine components. This study describes the quantitative characterization of in-process control (IPC) samples generated during the fermentation and purification of the capsular polysaccharide (CPS), polyribosyl-ribitol-phosphate (PRP), derived from Hib. Reliable quantitative methods are necessary for all stages of production; otherwise accurate process monitoring and validation is not possible. Prior to the availability of high performance anion exchange chromatography methods, this polysaccharide was predominantly quantified either with immunochemical methods, or with the colorimetric orcinol method, which shows interference from fermentation medium components and reagents used during purification. Next to an improved high performance anion exchange chromatography-pulsed amperometric detection (HPAEC-PAD) method, using a modified gradient elution, both the orcinol assay and high performance size exclusion chromatography (HPSEC) analyses were evaluated. For DSP samples, it was found that the correlation between the results obtained by HPAEC-PAD specific quantification of the PRP monomeric repeat unit released by alkaline hydrolysis, and those from the orcinol method was high (R(2)=0.8762), and that it was lower between HPAEC-PAD and HPSEC results. Additionally, HPSEC analysis of USP samples yielded surprisingly comparable results to those obtained by HPAEC-PAD. In the early part of the fermentation, medium components interfered with the different types of analysis, but quantitative HPSEC data could still be obtained, although lacking the specificity of the HPAEC-PAD method. Thus, the HPAEC

  15. Development of colorimetric solid phase extraction (C-SPE) for in-flight monitoring of spacecraft water supplies

    NASA Astrophysics Data System (ADS)

    Gazda, Daniel Bryan

    2004-12-01

    Colorimetric solid phase extraction (C-SPE) is a sorption-spectrophotometric technique that combines colorimetric reagents, solid phase extraction, and diffuse reflectance spectroscopy to quantify trace analytes in water samples. In C-SPE, a syringe is used to meter a known volume of sample through a membrane impregnated with a selective colorimetric reagent along with any additives required to optimize the complexation of the reagent and analyte. As the sample is passed through the membrane, analytes are extracted and complexed, leading to a detectable change in the optical characteristics of the membrane. The analyte-reagent complex is then quantified directly on the membrane, using a hand-held diffuse reflectance spectrophotometer. This dissertation focuses on the development, ground testing, and microgravity flight demonstration of C-SPE methods to meet the near- and long-term water quality monitoring needs of NASA. To this end, the ability of C-SPE to function in a microgravity environment was tested through performance evaluations of methods for the determination of the biocidal agents silver(I) and iodine on the KC-135 microgravity simulator. The biocidal iodine platform was investigated further to determine which iodine species is responsible for the C-SPE signal. Through systematic comparisons of C-SPE results and UV-Visible absorbance studies it was determined that biocidally active I2 is the iodine species complexed by poly(vinylpyrrolidone). The application of C-SPE to additional target water quality parameters is demonstrated through the determination of nickel(II), a metal leachate found in archived water samples from the International Space Station, using dimethylglyoxime. This method introduced a new variation of C-SPE, the quantification of trace analytes based on the collection of an insoluble, colored precipitate. The nickel(II) method was then combined with the method for biocidal silver(I) and a new method to measure sample pH to create a

  16. The rapid identification and quantification of iso-α-acids and reduced iso-α-acids in blood using UHPLC-MS/MS: validation of a novel marker for beer consumption.

    PubMed

    Rodda, Luke N; Gerostamoulos, Dimitri; Drummer, Olaf H

    2013-12-01

    A method for the detection of iso-α-acid (IAA) type ingredient congeners that are derived from the hop plant (Humulus lupulus L.) was developed to detect recent consumption of beer in blood. Three structurally similar but chemically altered IAA, also used as beer-specific ingredients, are known as "reduced IAA", consisting of the rho-, tetrahydro-, and hexahydro-IAA were also targeted. The use of a simple protein precipitation extraction and ultrahigh-performance liquid chromatography system coupled with a tandem mass spectrometer system enabled detection of these analytes in both antemortem and postmortem blood. Extracts were injected onto a C18 solid-core column under gradient elution to achieve separation of isobaric analogs and isomers within a 10-min run time. Electrospray ionization in negative multiple reaction monitoring mode was used to monitor three transitions for each of the analytes that were ultimately grouped together to form a calibration curve for quantification of each of the four IAA groups. The method was fully validated according to international guidelines that included extraction efficiency, matrix effects, process efficiency, ion suppression/enhancement of co-eluting analytes, selectivity, crosstalk, accuracy and precision, stabilities, and lower limits of quantification. Finally, applicability of the method described was demonstrated by the detection of IAA ingredient congeners in the blood of a volunteer following the consumption of a relatively small amount of beer in a pilot study. PMID:24177342

  17. Interfacial polymerization for colorimetric labeling of protein expression in cells.

    PubMed

    Lilly, Jacob L; Sheldon, Phillip R; Hoversten, Liv J; Romero, Gabriela; Balasubramaniam, Vivek; Berron, Brad J

    2014-01-01

    Determining the location of rare proteins in cells typically requires the use of on-sample amplification. Antibody based recognition and enzymatic amplification is used to produce large amounts of visible label at the site of protein expression, but these techniques suffer from the presence of nonspecific reactivity in the biological sample and from poor spatial control over the label. Polymerization based amplification is a recently developed alternative means of creating an on-sample amplification for fluorescence applications, while not suffering from endogenous labels or loss of signal localization. This manuscript builds upon polymerization based amplification by developing a stable, archivable, and colorimetric mode of amplification termed Polymer Dye Labeling. The basic concept involves an interfacial polymer grown at the site of protein expression and subsequent staining of this polymer with an appropriate dye. The dyes Evans Blue and eosin were initially investigated for colorimetric response in a microarray setting, where both specifically stained polymer films on glass. The process was translated to the staining of protein expression in human dermal fibroblast cells, and Polymer Dye Labeling was specific to regions consistent with desired protein expression. The labeling is stable for over 200 days in ambient conditions and is also compatible with modern mounting medium. PMID:25536421

  18. Paper bioassay based on ceria nanoparticles as colorimetric probes.

    PubMed

    Ornatska, Maryna; Sharpe, Erica; Andreescu, Daniel; Andreescu, Silvana

    2011-06-01

    We report the first use of redox nanoparticles of cerium oxide as colorimetric probes in bioanalysis. The method is based on changes in the physicochemical properties of ceria nanoparticles, used here as chromogenic indicators, in response to the analyte. We show that these particles can be fully integrated in a paper-based bioassay. To construct the sensor, ceria nanoparticles and glucose oxidase were coimmobilized onto filter paper using a silanization procedure. In the presence of glucose, the enzymatically generated hydrogen peroxide induces a visual color change of the ceria nanoparticles immobilized onto the bioactive sensing paper, from white-yellowish to dark orange, in a concentration-dependent manner. A detection limit of 0.5 mM glucose with a linear range up to 100 mM and a reproducibility of 4.3% for n = 11 ceria paper strips were obtained. The assay is fully reversible and can be reused for at least 10 consecutive measurement cycles, without significant loss of activity. Another unique feature is that it does not require external reagents, as all the sensing components are fixed onto the paper platform. The bioassay can be stored for at least 79 days at room temperature while maintaining the same analytical performance. An example of analytical application was demonstrated for the detection of glucose in human serum. The results demonstrate the potential of this type of nanoparticles as novel components in the development of robust colorimetric bioassays. PMID:21524141

  19. Interfacial Polymerization for Colorimetric Labeling of Protein Expression in Cells

    PubMed Central

    Lilly, Jacob L.; Sheldon, Phillip R.; Hoversten, Liv J.; Romero, Gabriela; Balasubramaniam, Vivek; Berron, Brad J.

    2014-01-01

    Determining the location of rare proteins in cells typically requires the use of on-sample amplification. Antibody based recognition and enzymatic amplification is used to produce large amounts of visible label at the site of protein expression, but these techniques suffer from the presence of nonspecific reactivity in the biological sample and from poor spatial control over the label. Polymerization based amplification is a recently developed alternative means of creating an on-sample amplification for fluorescence applications, while not suffering from endogenous labels or loss of signal localization. This manuscript builds upon polymerization based amplification by developing a stable, archivable, and colorimetric mode of amplification termed Polymer Dye Labeling. The basic concept involves an interfacial polymer grown at the site of protein expression and subsequent staining of this polymer with an appropriate dye. The dyes Evans Blue and eosin were initially investigated for colorimetric response in a microarray setting, where both specifically stained polymer films on glass. The process was translated to the staining of protein expression in human dermal fibroblast cells, and Polymer Dye Labeling was specific to regions consistent with desired protein expression. The labeling is stable for over 200 days in ambient conditions and is also compatible with modern mounting medium. PMID:25536421

  20. A new colorimetric method for the estimation of glycosylated hemoglobin.

    PubMed

    Nayak, S S; Pattabiraman, T N

    1981-02-01

    A new colorimetric method, based on the phenol sulphuric acid reaction of carbohydrates, is described for the determination of glycosylated hemoglobin. Hemolyzates were treated with 1 mol/l oxalic acid in 2 mol/l Hcl for 4 h at 100 degrees C, the protein was precipitated with trichloroacetic acid, and the free sugars and hydroxymethyl furfural in the protein free supernatant were treated with phenol and sulphuric acid to form the color. The new method is compared to the thiobarbituric acid method and the ion-exchange chromatographic method for the estimation of glycosylated hemoglobin in normals and diabetics. The increase in glycosylated hemoglobin in diabetic patients as estimated by the phenol-sulphuric acid method was more significant (P less than 0.001) than the increase observed by the thiobarbituric acid method (P less than 0.01). The correlation between the phenol-sulphuric acid method and the column method was better (r = 0.91) than the correlation between the thiobarbituric acid method and the column method (r = 0.84). No significant correlation between fasting and postprandial blood sugar level and glycosylated hemoglobin level as determined by the two colorimetric methods was observed in diabetic patients. PMID:7226519

  1. A colorimetric sensor array for detection of triacetone triperoxide vapor.

    PubMed

    Lin, Hengwei; Suslick, Kenneth S

    2010-11-10

    Triacetone triperoxide (TATP), one of the most dangerous primary explosives, has emerged as an explosive of choice for terrorists in recent years. Owing to the lack of UV absorbance, fluorescence, or facile ionization, TATP is extremely difficult to detect directly. Techniques that are able to detect generally require expensive instrumentation, need extensive sample preparation, or cannot detect TATP in the gas phase. Here we report a simple and highly sensitive colorimetric sensor for the detection of TATP vapor with semiquantitative analysis from 50 ppb to 10 ppm. By using a solid acid catalyst to pretreat a gas stream, we have discovered that a colorimetric sensor array of redox sensitive dyes can detect even very low levels of TATP vapor from its acid decomposition products (e.g., H(2)O(2)) with limits of detection (LOD) below 2 ppb (i.e., <0.02% of its saturation vapor pressure). Common potential interferences (e.g., humidity, personal hygiene products, perfume, laundry supplies, volatile organic compounds, etc.) do not generate an array response, and the array can also differentiate TATP from other chemical oxidants (e.g., hydrogen peroxide, bleach, tert-butylhydroperoxide, peracetic acid). PMID:20949933

  2. Optical fiber waveguide sensor for the colorimetric detection of ammonia

    NASA Astrophysics Data System (ADS)

    Schmitt, Katrin; Rist, Jonas; Peter, Carolin; Wöllenstein, Jürgen

    2011-06-01

    We present the development and characterization of a fiber-optic colorimetric gas sensor combined with the electronic circuitry for measurement control and RFID communication. The gas sensor detects ammonia using a 300 μm polyolefin fiber coated with a gas-sensitive polymer film. The spectral and time-dependent sensitivity of various polymer films was tested in transmission measurements. Light from a standard LED at λ = 590 nm was coupled into the polyolefin fiber through the front face. A prototype of the gas sensor with the direct coupling method was tested under realistic measurement conditions, i.e. battery-driven and in a completely autonomous mode. The sensor system showed good sensitivity to the ammonia concentrations and response times in the order of minutes. The achievable power consumption was below 100μW.The films contained the pH-sensitive dyes bromocresol purple or bromophenol blue embedded in either ethyl cellulose or polyvinyl butyral, and optionally tributyl phosphate as plasticizer. The bromophenol blue based films showed a strong reaction to ammonia, with saturation concentrations around 1000 ppm and response times of about 15 seconds to 100ppm. The colorimetric reaction was simulated using a simple kinetic model which was in good agreement with the experimental results.

  3. Interface engineering catalytic graphene for smart colorimetric biosensing.

    PubMed

    Liu, Meng; Zhao, Huimin; Chen, Shuo; Yu, Hongtao; Quan, Xie

    2012-04-24

    Herein a hybrid catalyst consisting of "naked" Au-NPs in situ grown on graphene sheets is engineered, which exhibits a synergetic effect in mimicking peroxidase at its interface, although free Au-NPs or graphene alone has very little activity. What is more, one of the unique features of our synergetic catalyst is that its interface can be reversibly switched from "inactive" to "active" upon treatment with different ssDNA species in solution, thus providing a powerful and versatile basis for designing graphene/DNA-based label-free colorimetric biosensors. Compared with other signal transduction modes in traditional graphene/aptamer-based systems, our novel signaling strategy not only avoids any labeling or modification procedures but also reduces the background signal due to the "off-on" switching mode during the sensing. Furthermore, this facile and general approach can be applicable to the other extended graphene/aptamer-based systems for colorimetric detection of a wide range of analytes. We envision that the tunable graphene-based smart interface could find potential applications in the development of biocatalysis, bioassays, and smart material devices in the future. PMID:22443302

  4. [Colorimetric characterization of LCD based on wavelength partition spectral model].

    PubMed

    Liu, Hao-Xue; Cui, Gui-Hua; Huang, Min; Wu, Bing; Xu, Yan-Fang; Luo, Ming

    2013-10-01

    To establish a colorimetrical characterization model of LCDs, an experiment with EIZO CG19, IBM 19, DELL 19 and HP 19 LCDs was designed and carried out to test the interaction between RGB channels, and then to test the spectral additive property of LCDs. The RGB digital values of single channel and two channels were given and the corresponding tristimulus values were measured, then a chart was plotted and calculations were made to test the independency of RGB channels. The results showed that the interaction between channels was reasonably weak and spectral additivity property was held well. We also found that the relations between radiations and digital values at different wavelengths varied, that is, they were the functions of wavelength. A new calculation method based on piecewise spectral model, in which the relation between radiations and digital values was fitted by a cubic polynomial in each piece of wavelength with measured spectral radiation curves, was proposed and tested. The spectral radiation curves of RGB primaries with any digital values can be found out with only a few measurements and fitted cubic polynomial in this way and then any displayed color can be turned out by the spectral additivity property of primaries at given digital values. The algorithm of this method was discussed in detail in this paper. The computations showed that the proposed method was simple and the number of measurements needed was reduced greatly while keeping a very high computation precision. This method can be used as a colorimetrical characterization model. PMID:24409730

  5. DNA transducer-triggered signal switch for visual colorimetric bioanalysis.

    PubMed

    Chen, Wenhong; Yan, Yurong; Zhang, Ye; Zhang, Xuemei; Yin, Yibing; Ding, Shijia

    2015-01-01

    A simple and versatile colorimetric biosensor has been developed for sensitive and specific detection of a wide range of biomolecules, such as oligonucleotides and aptamer-recognized targets. Combining the signal transducer and catalyzed hairpin assembly (CHA)-based signal amplification, the target DNA binds with the hairpin DNA to form a new nucleic acid sequence and creates a toehold in the transducer for initiating the recycle amplification reaction of CHA. The catalyzed assembly process produces a large amount of G-rich DNA. In the presence of hemin, the G-rich DNA forms G-quadruplex/hemin complex and mimic horseradish peroxidase activity, which catalyzes a colorimetric reaction. Under optimal conditions, the calibration curve of synthetic target DNA has good linearity from 50 pM to 200 nM with a detection limit of 32 pM. This strategy has been successfully applied to detect S. pneumoniae as low as 156 CFU mL(-1), and shows a good specificity against closely related streptococci and major pathogenic bacteria. In addition, the developed method enables successful visual analysis of S. pneumoniae in clinical samples by the naked eye. Importantly, this method demonstrates excellent assay versatility for sensitively detecting oligonucleotides or aptamer-recognized targets. PMID:26060886

  6. Colorimetric logic response based on aptamer functionalized colloidal crystal hydrogels

    NASA Astrophysics Data System (ADS)

    Ye, Baofen; Wang, Huan; Ding, Haibo; Zhao, Yuanjin; Pu, Yuepu; Gu, Zhongze

    2015-04-01

    A novel colorimetric logic system based on the aptamer-cross-linked colloidal crystal hydrogel (CCH) was developed. With the input stimuli of Hg2+ and Ag+, the CCH displayed shrinking response and colour change corresponding to the logical ``OR'' and ``AND'' gate. The visualization of the logic output signals is realized.A novel colorimetric logic system based on the aptamer-cross-linked colloidal crystal hydrogel (CCH) was developed. With the input stimuli of Hg2+ and Ag+, the CCH displayed shrinking response and colour change corresponding to the logical ``OR'' and ``AND'' gate. The visualization of the logic output signals is realized. Electronic supplementary information (ESI) available: I. Experimental section. II. Photograph of the aptamer functionalized CCH in the presence of different targets. III. The specificity of the aptamer functionalized CCH. IV. Relationship between the input ion concentration and the reflection wavelength blue shift. V. The logic swelling kinetics of CCH. See DOI: 10.1039/c5nr00586h

  7. Colorimetric paper bioassay for the detection of phenolic compounds.

    PubMed

    Alkasir, Ramiz S J; Ornatska, Maryna; Andreescu, Silvana

    2012-11-20

    A new type of paper based bioassay for the colorimetric detection of phenolic compounds including phenol, bisphenol A, catechol and cresols is reported. The sensor is based on a layer-by-layer (LbL) assembly approach formed by alternatively depositing layers of chitosan and alginate polyelectrolytes onto filter paper and physically entrapping the tyrosinase enzyme in between these layers. The sensor response is quantified as a color change resulting from the specific binding of the enzymatically generated quinone to the multilayers of immobilized chitosan on the paper. The color change can be quantified with the naked eye but a digitalized picture can also be used to provide more sensitive comparison to a calibrated color scheme. The sensor was optimized with respect to the number of layers, pH, enzyme, chitosan and alginate amounts. The colorimetric response was concentration dependent, with a detection limit of 0.86 (±0.1) μg/L for each of the phenolic compounds tested. The response time required for the sensor to reach steady-state color varied between 6 and 17 min depending on the phenolic substrate. The sensor showed excellent storage stability at room temperature for several months (92% residual activity after 260 days storage) and demonstrated good functionality in real environmental samples. A procedure to mass-produce the bioactive sensors by inkjet printing the LbL layers of polyelectrolyte and enzyme on paper is demonstrated. PMID:23113670

  8. Sensitive and visual detection of sequence-specific DNA-binding protein via a gold nanoparticle-based colorimetric biosensor.

    PubMed

    Ou, Li-Juan; Jin, Pei-Yan; Chu, Xia; Jiang, Jian-Hui; Yu, Ru-Qin

    2010-07-15

    A novel exonuclease III (Exo III) protection-based colorimetric biosensing strategy was developed for rapid, sensitive, and visual detection of sequence-specific DNA-binding proteins. This strategy relied on the protection of DNA-cross-linked gold nanoparticle (AuNP) aggregates from Exo III-mediated digestion by specific interactions of target proteins with their binding sequences. Interestingly, we disclosed a new finding that binding of target proteins to their binding sequences in the aggregated AuNP network rendered a stable and long-period protection of DNA. Unlike conventional fluorescence assays merely based on temporal protection of DNA from Exo III digestion, the stable protection afforded a static color transition indicator for DNA-protein interactions with no time-dependent monitoring required in the assay. Therefore, it furnished the developed strategy with improved technical robustness and operational convenience. Furthermore, we introduced thioctic acid as a stable anchor for tethering DNA on AuNPs. This DNA-tethering protocol circumvented the interferences from thiol compounds in common enzymatic systems. The Exo III protection-based colorimetric biosensor was demonstrated using a model target of TATA binding protein, a key transcriptional factor involving in various transcriptional regulatory networks. The results revealed that the method allowed a specific, simple, and quantitative assay of the target protein with a linear response range from 0 to 120 nM and a detection limit of 10 nM. PMID:20565105

  9. A Colorimetric Microplate Assay for DNA-Binding Activity of His-Tagged MutS Protein.

    PubMed

    Banasik, Michał; Sachadyn, Paweł

    2016-09-01

    A simple microplate method was designed for rapid testing DNA-binding activity of proteins. The principle of the assay involves binding of tested DNA by his-tagged protein immobilized on a nickel-coated ELISA plate, following colorimetric detection of biotinylated DNA with avidin conjugated to horseradish peroxidase. The method was used to compare DNA mismatch binding activities of MutS proteins from three bacterial species. The assay required relatively low amounts of tested protein (approximately 0.5-10 pmol) and DNA (0.1-10 pmol) and a relatively short time of analysis (up to 60 min). The method is very simple to apply and convenient to test different buffer conditions of DNA-protein binding. Sensitive colorimetric detection enables naked eye observations and quantitation with an ELISA reader. The performance of the assay, which we believe is a distinguishing trait of the method, is based on two strong and specific molecular interactions: binding of a his-tagged protein to a nickel-coated microplate and binding of biotinylated DNA to avidin. In the reported experiments, the solution was used to optimize the conditions for DNA mismatch binding by MutS protein; however, the approach could be implemented to test nucleic acids interactions with any protein of interest. PMID:27241123

  10. Colorimetric Evaluation of the Viability of the Microalga Dunaliella Salina as a Test Tool for Nanomaterial Toxicity.

    PubMed

    Golubev, Alexander A; Prilepskii, Artur Y; Dykman, Lev A; Khlebtsov, Nikolai G; Bogatyrev, Vladimir A

    2016-05-01

    A diagnostic test system was developed to determine the toxicity of nanomaterials to the saltwater microalga Dunaliella salina through evaluation of cell death and changes in the culture growth rate at various toxicant concentrations, providing LC50 and other toxicological metrics. The viability of cells was shown to decrease with decreasing chlorophyll absorption of red light by damaged cells. This correlation was confirmed by independent fluorescence microscopic measurements of live and dead cells in the population. Two standard colorless pollutants, hydrogen peroxide and formaldehyde, were used to validate the colorimetric method. The method's performance is exemplified with three Ag-containing preparations (Ag nitrate, Ag proteinate, and 20-nm Ag nanoparticles) and with cetyltrimethylammonium bromide (CTAB) mixed with colloidal 15-nm Au and 20-nm Ag nanoparticles. The toxicity of the Ag-containing preparations to D. salina decreased in the order Ag nitrate ≥ Ag proteinate ≫ colloidal Ag. The toxicity of colloidal Au-CTAB mixtures was found to depend mostly on the content of free CTAB. The toxicity of colloidal Ag increased substantially in the presence of CTAB. The results suggest that our D. salina-based colorimetric test system can be used for simple and rapid preliminary screening of the toxicity of different nanomaterials. PMID:26865664

  11. Optimizing Colorimetric Assay Based on V2O5 Nanozymes for Sensitive Detection of H2O2 and Glucose

    PubMed Central

    Sun, Jiaheng; Li, Chunyan; Qi, Yanfei; Guo, Shuanli; Liang, Xue

    2016-01-01

    Nanozyme-based chemical sensing is a rapidly emerging field of research. Herein, a simple colorimetric assay for the detection of hydrogen peroxide and glucose based on the peroxidase-like activity of V2O5 nanozymes has been established. In this assay, the effects of pH, substrate, nanozyme concentrations and buffer solution have been investigated. It was found that compared with 3,3′,5,5′-tetramethylbenzidine (TMB), the enzyme substrate o-phenylenediamine (OPD) seriously interfered with the H2O2 detection. Under the optimal reaction conditions, the resulting sensor displayed a good response to H2O2 with a linear range of 1 to 500 μM, and a detection limit of 1 μM at a signal-to-noise ratio of 3. A linear correlation was established between absorbance intensity and concentration of glucose from 10 to 2000 μM, with a detection limit of 10 μM. The current work presents a simple, cheap, more convenient, sensitive, and easy handling colorimetric assay. PMID:27110794

  12. A simple highly sensitive and selective aptamer-based colorimetric sensor for environmental toxins microcystin-LR in water samples.

    PubMed

    Li, Xiuyan; Cheng, Ruojie; Shi, Huijie; Tang, Bo; Xiao, Hanshuang; Zhao, Guohua

    2016-03-01

    A simple and highly sensitive aptamer-based colorimetric sensor was developed for selective detection of Microcystin-LR (MC-LR). The aptamer (ABA) was employed as recognition element which could bind MC-LR with high-affinity, while gold nanoparticles (AuNPs) worked as sensing materials whose plasma resonance absorption peaks red shifted upon binding of the targets at a high concentration of sodium chloride. With the addition of MC-LR, the random coil aptamer adsorbed on Au NPs altered into regulated structure to form MC-LR-aptamer complexes and broke away from the surface of Au NPs, leading to the aggregation of AuNPs, and the color converted from red to blue due to the interparticle plasmon coupling. Results showed that our aptamer-based colorimetric sensor exhibited rapid and sensitive detection performance for MC-LR with linear range from 0.5 nM to 7.5 μM and the detection limit reached 0.37 nM. Meanwhile, the pollutants usually coexisting with MC-LR in pollutant water samples had not demonstrated disturbance for detecting of MC-LR. The mechanism was also proposed suggesting that high affinity interaction between aptamer and MC-LR significantly enhanced the sensitivity and selectivity for MC-LR detection. Besides, the established method was utilized in analyzing real water samples and splendid sensitivity and selectivity were obtained as well. PMID:26619046

  13. Optimizing Colorimetric Assay Based on V₂O₅ Nanozymes for Sensitive Detection of H₂O₂ and Glucose.

    PubMed

    Sun, Jiaheng; Li, Chunyan; Qi, Yanfei; Guo, Shuanli; Liang, Xue

    2016-01-01

    Nanozyme-based chemical sensing is a rapidly emerging field of research. Herein, a simple colorimetric assay for the detection of hydrogen peroxide and glucose based on the peroxidase-like activity of V₂O₅ nanozymes has been established. In this assay, the effects of pH, substrate, nanozyme concentrations and buffer solution have been investigated. It was found that compared with 3,3',5,5'-tetramethylbenzidine (TMB), the enzyme substrate o-phenylenediamine (OPD) seriously interfered with the H₂O₂ detection. Under the optimal reaction conditions, the resulting sensor displayed a good response to H₂O₂ with a linear range of 1 to 500 μM, and a detection limit of 1 μM at a signal-to-noise ratio of 3. A linear correlation was established between absorbance intensity and concentration of glucose from 10 to 2000 μM, with a detection limit of 10 μM. The current work presents a simple, cheap, more convenient, sensitive, and easy handling colorimetric assay. PMID:27110794

  14. A colorimetric probe based on desensitized ionene-stabilized gold nanoparticles for single-step test for sulfate ions.

    PubMed

    Arkhipova, Viktoriya V; Apyari, Vladimir V; Dmitrienko, Stanislava G

    2015-03-15

    Desensitized ionene-stabilized gold nanoparticles have been prepared and applied as a colorimetric probe for the single-step test for sulfate ions at the relatively high concentration level. The approach is based on aggregation of the nanoparticles leading to the change in absorption spectra and color of the solution. These nanoparticles are characterized by the decreased sensitivity due to both electrostatic and steric stabilization, which allows for simple, and rapid direct single-step determination of sulfate at the relatively high concentration level in real water samples without sample pretreatment or dilution. Influence of different factors (the time of interaction, pH, the concentrations of sulfate ions and the nanoparticles) on the aggregation and analytical performance of the procedure was investigated. The method allows for the determination of sulfate ions in the mass range of 0.2-0.4 mg with RSD of 5% from the sample volume of less than 2 mL. It has a sharp dependence of the colorimetric response on the concentration of sulfate, which makes it prospective for indicating deviations of the sulfate concentration regarding some declared value chosen within the above range. The time of the analysis is 2 min. The method was applied to the analysis of mineral water samples. PMID:25574653

  15. Atomic absorption spectroscopic, conductometric and colorimetric methods for determination of fluoroquinolone antibiotics using ammonium reineckate ion-pair complex formation

    NASA Astrophysics Data System (ADS)

    Ragab, Gamal H.; Amin, Alaa S.

    2004-03-01

    Three accurate, rapid and simple atomic absorption spectrometric, conductometric and colorimetric methods were developed for the determination of norfloxacin (NRF), ciprofloxacin (CIP), ofloxacin (OFL) and enrofloxacin (ENF). The proposed methods depend upon the reaction of ammonium reineckate with the studied drugs to form stable precipitate of ion-pair complexes, which was dissolved in acetone. The pink coloured complexes were determined either by AAS or colorimetrically at λmax 525 nm directly using the dissolved complex. Using conductometric titration, the studied drugs could be evaluated in 50% (v/v) acetone in the range 5.0-65, 4.0-48, 5.0-56 and 6.0-72 μg ml -1 of NRF, CPF, OFL and ENF, respectively. The optimizations of various experimental conditions were described. The results obtained showed good recoveries of 99.15±1.15, 99.30±1.40, 99.60±1.50, and 99.00±1.25% with relative standard deviations of 0.81, 1.06, 0.97, and 0.69% for NRF, CPF, OFL, and ENF, respectively. Applications of the proposed methods to representative pharmaceutical formulations are successfully presented.

  16. A colorimetric biosensor for detection of attomolar microRNA with a functional nucleic acid-based amplification machine.

    PubMed

    Li, Dandan; Cheng, Wei; Yan, Yurong; Zhang, Ye; Yin, Yibing; Ju, Huangxian; Ding, Shijia

    2016-01-01

    A functional nucleic acid-based amplification machine was designed for simple and label-free ultrasensitive colorimetric biosensing of microRNA (miRNA). The amplification machine was composed of a complex of trigger template and C-rich DNA modified molecular beacon (MB) and G-rich DNA (GDNA) as the probe, polymerase and nicking enzyme, and a dumbbell-shaped amplification template. The presence of target miRNA triggered MB mediated strand displacement to cyclically release nicking triggers, which led to a toehold initiated rolling circle amplification to produce large amounts of GDNAs. The formed GDNAs could stack with hemin to form G-quadruplex/hemin DNAzyme, a well-known horseradish peroxidase (HRP) mimic, for catalyzing a colorimetric reaction. The modified MB improved the stringent target recognition and reduced background signal. The proposed sensing strategy showed very high sensitivity and selectivity with a wide dynamic range from 10 aM to 1.0 nM, and enabled successful visual analysis of trace amount of miRNA in real sample by the naked eye. This rapid and highly efficient signal amplification strategy provided a simple and sensitive platform for miRNA detection. It would be a versatile and powerful tool for clinical molecular diagnostics. PMID:26695292

  17. Colorimetric detection of Mn2+ using silver nanoparticles cofunctionalized with 4-mercaptobenzoic acid and melamine as a probe.

    PubMed

    Zhou, Ying; Zhao, Hong; Li, Chang; He, Peng; Peng, Wenbo; Yuan, Longfei; Zeng, Lixi; He, Yujian

    2012-08-15

    A facile, selective and highly sensitive method is proposed for colorimetric detection of manganese ions using 4-mercaptobenzoic acid (4-MBA) and melamine (MA) modified silver nanoparticles (AgNPs). The presence of Mn(2+) induces the aggregation of AgNPs through cooperative metal-ligand interaction, resulting in a color change from bright yellow to purple. The cofunctionalized AgNPs showed obvious advantages over the ones functionalized only by 4-MBA or MA in terms of selectivity. Mn(2+) could be monitored by colorimetric response of AgNPs by a UV-vis spectrophotometer or even naked eyes. The absorbance ratio (A(550 nm)/A(408 nm)) is linear with the concentration of Mn(2+) ranging from 5×10(-7) mol/L to 1×10(-5) mol/L with a correlation coefficient of 0.993, and the detection limit is as low as 5×10(-8) mol/L. Particularly, this cost-effective process also allowed rapid and simple determination of the Mn(2+) in drinking water. PMID:22841088

  18. A dual-mode colorimetric and fluorometric "light on" sensor for thiocyanate based on fluorescent carbon dots and unmodified gold nanoparticles.

    PubMed

    Zhao, Dan; Chen, Chuanxia; Lu, Lixia; Yang, Fan; Yang, Xiurong

    2015-12-21

    A novel, highly sensitive and selective dual-readout (colorimetric and fluorometric) sensor based on fluorescent carbon dots (CDs) and unmodified gold nanoparticles (AuNPs) for the detection of thiocyanate (SCN(-)) was proposed. Amino-functionalized CDs could be readily adsorbed onto the surface of citrate-stabilized AuNPs through Au-N interactions, leading to the aggregation of AuNPs and the nonfluorescent off-state of CDs arising from potential fluorescence resonance energy transfer (FRET). However, SCN(-) had a stronger affinity toward AuNPs and could compete with CDs to bind onto the surface of AuNPs in priority, which prevented the aggregation of AuNPs and fluorescence quenching of CDs. Correspondingly, both the colorimetric and fluorometric signals remained "light-on". The color of the sensing solution remained red and the fluorescence remained unquenched. A distinguishable change in the color was observed at a SCN(-) concentration of 1 μM by the naked eye and a detection limit as low as 0.036 μM was obtained by virtue of fluorescence spectroscopy. Both colorimetric and fluorometric sensors exhibited excellent selectivity toward SCN(-) over other common metallic ions and anions. In addition, such a sensing assay featured simplicity, rapidity, cost-effectiveness and ease of operation without further modification. The accuracy and precision were evaluated based on the quantitative detection of SCN(-) in tap water and saliva samples with satisfactory results. PMID:26567774

  19. Loop-mediated isothermal amplification for rapid and semiquantitative detection of Loa loa infection.

    PubMed

    Drame, Papa M; Fink, Doran L; Kamgno, Joseph; Herrick, Jesica A; Nutman, Thomas B

    2014-06-01

    Rapid and accurate tests are currently needed to identify individuals with high levels of Loa loa microfilaria (mf), so that these individuals may be excluded from mass ivermectin administration campaigns against onchocerciasis and lymphatic filariasis being conducted in areas where Onchocerca volvulus, Wuchereria bancrofti, and L. loa are coendemic. To address this need, colorimetric loop-mediated isothermal amplification (LAMP) assays targeting the L. loa-specific gene sequences LLMF72 and LLMF342 were developed for the detection and quantification of L. loa microfilaremia. Both LAMP assays were highly specific (100%) for L. loa infection compared to the absence of infection or infection with related filarial pathogens. The LLMF72-based LAMP assay showed greater analytic sensitivity (limit of detection, 0.1 pg/ml of genomic DNA [gDNA] and/or 5 mf/ml) than the LLMF342-based LAMP assay (10 pg/ml of gDNA and/or 50 mf/ml), and its analytic sensitivity was similar to that of LLMF72-based quantitative PCR (qPCR). A high level of correlation was observed between microfilaria counts as determined by LLMF72-based qPCR and time to positivity by the LAMP assay, and performance measures of sensitivity, specificity, and positive and negative predictive values were similar for both assays when applied to field-collected clinical samples. By simply varying the run time, the LAMP assay was able to accurately distinguish individuals at risk for serious adverse events (SAEs) after exposure to ivermectin, using thresholds of >5,000 mf/ml and >30,000 mf/ml as indicators of increasing levels of risk. In summary, LLMF72 LAMP represents a new molecular diagnostic tool that is readily applicable as a point-of-care method for L. loa microfilarial detection and quantification in resource-limited countries where L. loa infection is endemic. PMID:24696020

  20. BacT/Alert: an automated colorimetric microbial detection system.

    PubMed Central

    Thorpe, T C; Wilson, M L; Turner, J E; DiGuiseppi, J L; Willert, M; Mirrett, S; Reller, L B

    1990-01-01

    BacT/Alert (Organon Teknika Corp., Durham, N.C.) is an automated microbial detection system based on the colorimetric detection of CO2 produced by growing microorganisms. Results of an evaluation of the media, sensor, detection system, and detection algorithm indicate that the system reliably grows and detects a wide variety of bacteria and fungi. Results of a limited pilot clinical trial with a prototype research instrument indicate that the system is comparable to the radiometric BACTEC 460 system in its ability to grow and detect microorganisms in blood. On the basis of these initial findings, large-scale clinical trials comparing BacT/Alert with other commercial microbial detection systems appear warranted. PMID:2116451

  1. Colorimetric and fluorometric dual-readout sensor for lysozyme.

    PubMed

    Zheng, Hanye; Qiu, Suyan; Xu, Kefeng; Luo, Linguang; Song, Yibiao; Lin, Zhenyu; Guo, Longhua; Qiu, Bin; Chen, Guonan

    2013-11-01

    A novel, highly sensitive and selective dual-readout sensor (colorimetric and fluorometric) for the detection of lysozyme was proposed. The fluorescence of triazolylcoumarin molecules was quenched by Au nanoparticles (AuNPs) initially through the fluorescence resonance energy transfer (FRET), after the addition of lysozyme, the stronger binding of lysozyme onto the surfaces of AuNPs made triazolylcoumarin molecules remove from the AuNPs surface and led to the recovery of the fluorescence of triazolylcoumarin molecules, and accompanied by the discernable color change of the solution from red to purple. The lowest detectable concentration for lysozyme was 50 ng mL(-1) by the naked eye, and the limit of detection (LOD) was 23 ng mL(-1) by fluorescence measurements. In addition, satisfactory results for lysozyme detection in hen egg white were confirmed in the study. Moreover, the presented sensor provides a reliable option to determine lysozyme with high sensitivity and selectivity. PMID:23978821

  2. Comparison of spectral colorimetric measurements vs. color pictures in dermatology

    NASA Astrophysics Data System (ADS)

    Blain, P.; Michel, F.; Moreau, V.,; Vanhooteghem, O.; Renotte, Y.; Habraken, S.

    2010-04-01

    We studied scars and wounds depths and surfaces thanks to our interferometric fringes projector 3D scanner1, 2. Color information of a wound indicates its deterioration level. That's why the visual color restitution, as realistic as possible, is a highly important parameter. Firstly our acquired 3D pictures were color mapped with an image recorded by a RGB camera. The results were not efficient enough. In order to improve our technique and provide more precise information, we add a spectral characterization to the set-up. Before adding the spectral information and a realistic color mapping to the 3D measurements, we evaluate the performances of colorimetric measurements. The tests have been made on mice with scars on their back.

  3. Simple colorimetric method for seminal plasma zinc assay.

    PubMed

    Fuentes, J; Miró, J; Riera, J

    1982-01-01

    Zinc concentration in seminal plasma is one of the best biochemical parameters with which to assess prostatic function in the study of male infertility. Nevertheless, this parameter is not determined in many laboratories because of technical difficulties. In this paper a simple colorimetric method for seminal plasma zinc assay is presented. This assay is performed with only one reagent solution making deproteinization and extraction with organic solvents unnecessary. The method is based on the reaction between 1-(2-pyridylazo)-2-naphthol and zinc which yields a coloured complex, that is soluble in the reagent solution when dodecyl hydrogen sulfate sodium salt is present. The solubilized complex has an absorbance peak at 560 nm. The accuracy of this method is proved by statistical comparison with atomic absorption spectrophotometry. Furthermore, the method has a good precision and is free of interference. PMID:7125243

  4. Simple colorimetric method for determination of thiamine hydrochloride in pharmaceuticals.

    PubMed

    Sane, R T; Doshi, V J; Jukar, S; Joshi, S K; Sawant, S V; Pandit, U R

    1985-01-01

    A simple colorimetric method is described for the determination of thiamine hydrochloride (vitamin B1) in dosage forms. The method is based on measurement of a yellow complex formed when thiamine HCl is treated with p-methylaminophenol sulfate (Metol) under alkaline conditions. Compounds such as vitamins A, B2, B6, B12, C, D, and E, and niacinamide, citric acid, liquid glucose, calcium pantothenate, biotin, liver extract, and folic acid do not interfere in the reaction. Extracting the complex into chloroform before quantitation enhances the stability of the reaction product and removes interference of water-soluble colored constituents in syrup samples. Statistical validation shows that the method is precise and accurate. Results agree well with those obtained by other methods in the literature. PMID:3980419

  5. Scoliosis quantification: an overview

    PubMed Central

    Kawchuk, Greg; McArthur, Ross

    1997-01-01

    Scoliotic curvatures have long been a focus of attention for clinicians and research scientists alike. The study, treatment and ultimately, the prevention of this prevalent health condition are impeded by the absence of an accurate, reliable, convenient and safe method of scoliosis quantification. The purpose of this paper is to provide an overview of the current methods of scoliosis quantification for clinicians who address this condition in their practices.

  6. Colorimetric analysis of outdoor illumination across varieties of atmospheric conditions.

    PubMed

    Peyvandi, Shahram; Hernández-Andrés, Javier; Olmo, F J; Nieves, Juan Luis; Romero, Javier

    2016-06-01

    Solar illumination at ground level is subject to a good deal of change in spectral and colorimetric properties. With an aim of understanding the influence of atmospheric components and phases of daylight on colorimetric specifications of downward radiation, more than 5,600,000 spectral irradiance functions of daylight, sunlight, and skylight were simulated by the radiative transfer code, SBDART [Bull. Am. Meteorol. Soc.79, 2101 (1998)], under the atmospheric conditions of clear sky without aerosol particles, clear sky with aerosol particles, and overcast sky. The interquartile range of the correlated color temperatures (CCT) for daylight indicated values from 5712 to 7757 K among the three atmospheric conditions. A minimum CCT of ∼3600  K was found for daylight when aerosol particles are present in the atmosphere. Our analysis indicated that hemispheric daylight with CCT less than 3600 K may be observed in rare conditions in which the level of aerosol is high in the atmosphere. In an atmosphere with aerosol particles, we also found that the chromaticity of daylight may shift along the green-purple direction of the Planckian locus, with a magnitude depending on the spectral extinction by aerosol particles and the amount of water vapor in the atmosphere. The data analysis showed that an extremely high value of CCT, in an atmosphere without aerosol particles, for daylight and skylight at low sun, is mainly due to the effect of Chappuis absorption band of ozone at ∼600  nm. In this paper, we compare our data with well-known observations from previous research, including the ones used by the CIE to define natural daylight illuminants. PMID:27409431

  7. Ink-jet printed colorimetric gas sensors on plastic foil

    NASA Astrophysics Data System (ADS)

    Courbat, Jerome; Briand, Danick; de Rooij, Nico F.

    2010-08-01

    An all polymeric colorimetric gas sensor with its associated electronics for ammonia (NH3) detection targeting low-cost and low-power applications is presented. The gas sensitive layer was inkjet printed on a plastic foil. The use of the foil directly as optical waveguide simplified the fabrication, made the device more cost effective and compatible with large scale fabrication techniques, such as roll to roll processes. Concentrations of 500 ppb of NH3 in nitrogen with 50% of RH were measured with a power consumption of about 868 μW in an optical pulsed mode of operation. Such sensors foresee applications in the field of wireless systems, for environmental and safety monitoring. The fabrication of the planar sensor was based on low temperature processing. The waveguide was made of PEN or PET foil and covered with an ammonia sensitive layer deposited by inkjet printing, which offered a proper and localized deposition of the film. The influence of the substrate temperature and its surface pretreatment were investigated to achieve the optimum deposition parameters for the printed fluid. To improve the light coupling from the light source (LED) to the detectors (photodiodes), polymeric micro-mirrors were patterned in an epoxy resin. With the printing of the colorimetric film and additive patterning of polymeric micro-mirrors on plastic foil, a major step was achieved towards the implementation of full plastic selective gas sensors. The combination with printed OLED and PPD would further lead to an integrated all polymeric optical transducer on plastic foil fully compatible with printed electronics processes.

  8. A sensitive and label-free trypsin colorimetric sensor with cytochrome c as a substrate.

    PubMed

    Zhang, Lufeng; Du, Jianxiu

    2016-05-15

    The development of simple and sensitive methods for protease sensing plays important roles in clinical diagnostics and drug development. Here a simple, rapid, label-free, and sensitive trypsin colorimetric sensor was developed by employing cytochrome c (cyt c) as an enzyme substrate and 3,3´,5,5´-tetramethylbenzidine (TMB) as a chromogenic reagent. It was found that cyt c hardly catalyzes H2O2-mediated TMB oxidation to produce a blue solution. But the hydrolysate of cyt c by trypsin displays an intense catalytic effect on the aforementioned reaction, resulting in the formation of a blue solution immediately. The detection process allows visually perceiving as low as 50 ng/mL trypsin with the naked eyes. With the aid of a spectrophotometer, the absorbance at 652 nm was proportional to the concentration of trypsin in the range from 5.0 ng/mL to 2.0 μg/mL with a detection limit of 4.5 ng/mL. The sensor showed better precision with relative standard deviation of 2.5% and 1.7% for eleven repetitive measurements of 50.0 ng/mL and 1.0 μg/mL trypsin solution, respectively. The procedure has been successfully applied to the determination of trypsin in human urines and for inhibitor screening, demonstrating its potential application in clinic diagnosis and drug development. PMID:26724537

  9. Colorimetric recognition of pazufloxacin mesilate based on the aggregation of gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Kong, Sumei; Liao, Ming; Gu, Yu; Li, Nan; Wu, Pinping; Zhang, Tingting; He, Hua

    2016-03-01

    A novel colorimetric nanomaterial-assisted optical sensor for pazufloxacin mesilate was proposed for the first time. Pazufloxacin mesilate could induce the aggregation of glucose-reduced gold nanoparticles (AuNPs) through hydrogen-bonding interaction and electrostatic attraction, leading to the changes in color and absorption spectra of AuNPs. The effect of different factors such as pH, the amount of AuNPs, reaction time and reaction temperature was inspected. Under the optimum condition, UV-vis spectra showed that the absorption ratio (A670/A532) was linear with the concentration of pazufloxacin mesilate in the range from 9 × 10- 8 mol L- 1 to 7 × 10- 7 mol L- 1 with a linear coefficient of 0.9951. This method can be applied to detecting pazufloxacin mesilate with an ultralow detection limit of 7.92 × 10- 9 mol L- 1 without any complicated instruments. Through inspecting other analytes and ions, the anti-interference performance of AuNP detection system for pazufloxacin mesilate was excellent. For its high efficiency, rapid response rate as well as wide linear range, it had been successfully used to the analysis of pazufloxacin mesilate in human urine quantificationally.

  10. Colorimetric recognition of pazufloxacin mesilate based on the aggregation of gold nanoparticles.

    PubMed

    Kong, Sumei; Liao, Ming; Gu, Yu; Li, Nan; Wu, Pinping; Zhang, Tingting; He, Hua

    2016-03-15

    A novel colorimetric nanomaterial-assisted optical sensor for pazufloxacin mesilate was proposed for the first time. Pazufloxacin mesilate could induce the aggregation of glucose-reduced gold nanoparticles (AuNPs) through hydrogen-bonding interaction and electrostatic attraction, leading to the changes in color and absorption spectra of AuNPs. The effect of different factors such as pH, the amount of AuNPs, reaction time and reaction temperature was inspected. Under the optimum condition, UV-vis spectra showed that the absorption ratio (A670/A532) was linear with the concentration of pazufloxacin mesilate in the range from 9×10(-8) mol L(-1) to 7×10(-7) mol L(-1) with a linear coefficient of 0.9951. This method can be applied to detecting pazufloxacin mesilate with an ultralow detection limit of 7.92×10(-9) mol L(-1) without any complicated instruments. Through inspecting other analytes and ions, the anti-interference performance of AuNP detection system for pazufloxacin mesilate was excellent. For its high efficiency, rapid response rate as well as wide linear range, it had been successfully used to the analysis of pazufloxacin mesilate in human urine quantificationally. PMID:26774816

  11. DNAzyme-based colorimetric sensing of lead (Pb2+) using unmodified gold nanoparticle probes

    NASA Astrophysics Data System (ADS)

    Wei, Hui; Li, Bingling; Li, Jing; Dong, Shaojun; Wang, Erkang

    2008-03-01

    Novel functional oligonucleotides, especially DNAzymes with RNA-cleavage activity, have been intensively studied due to their potential applications in therapeutics and sensors. Taking advantage of the high specificity of 17E DNAzyme for Pb2+, highly sensitive and selective fluorescent, electrochemical and colorimetric sensors have been developed for Pb2+. In this work, we report a simple, sensitive and label-free 17E DNAzyme-based sensor for Pb2+ detection using unmodified gold nanoparticles (GNPs) based on the fact that unfolded single-stranded DNA could be adsorbed on the citrate protected GNPs while double-stranded DNA could not. By our method the substrate cleavage by the 17E DNAzyme in the presence of Pb2+ could be monitored by color change of GNPs, thereby Pb2+ detection was realized. The detection of Pb2+ could be realized within 20 min, with a detection limit of 500 nM. The selectivity of our sensor has been investigated by challenging the sensing system with other divalent metal ions. Since common steps such as modification and separation could be successfully avoided, the sensor developed here could provide a simple, cost-effective yet rapid and sensitive measurement tool for Pb2+ detection and may prove useful in the development of sensors for clinical toxicology and environmental monitoring in the future.

  12. Studies of a colorimetric array consisted of metalloporphrins and Pt complexes

    NASA Astrophysics Data System (ADS)

    Luo, Jing; Ma, Tengshuang; Li, Lu; Pan, Pinghua; Xu, Jianhua; Jiang, Yadong; Yang, Yajie

    2010-10-01

    We have studied the absorption of volatile organic compounds (VOCs) and vapochromic characterization by a colorimetric array based on metalloporphyrins and Pt complexes: ZnTPP, CuTPP, and [Pt(Me2bizmpy)Cl]Cl (TPP=tetraphenylporphrins; Me2bzimpy=2,6-bis(N-methylbenzimidazol-2-yl)pyridine). The array was exposed to pyridine, tetrahydrofuran, ethyl acetate, methanol and acetonitrile. The experimental results demonstrate that the Pt complexes are sensitive to methanol and acetonitrile, while ZnTPP and CuTPP present an obvious response to pyridine, tetrahydrofuran and ethyl acetate. The response behaviors are reversible and rapid within seconds. The changes of Ultra Violet Visible (UV-visible) absorption spectrums of ZnTPP and CuTPP LB films before and after exposure upon the VOCs are also discussed. In addition, the differences of absorption spectrum of Pt complexes dissolved in several solvents are mentioned. The results show that [Pt(Me2bizmpy)Cl]Cl, CuTPP and ZnTPP exhibit reversible changes of action with VOCs, and they show promising future to be used in Smell-Seeing Electronic Nose to identify VOCs.

  13. Novel core etching technique of gold nanoparticles for colorimetric dopamine detection.

    PubMed

    Lee, Ho-Cheng; Chen, Tzu-Heng; Tseng, Wei-Lung; Lin, Che-Hsin

    2012-11-21

    This study develops a novel and high performance colorimetric probe for dopamine (DA) detection. Aqueous-phase gold nanoparticles (AuNPs) extracted with 4-(dimethylamino)pyridine (DMAP) from toluene solvent are used as the reaction probes. The original AuNPs of diameter around 13 nm separate into 2-5 nm sizes when dopamine (DA) is added, resulting in the color change of the AuNP solution from red to blackish green. Transmission electron microscopy (TEM) observations and dynamic light scattering (DLS) tests show that the AuNPs break into their smaller sizes right after addition of DA. The results confirm that the DMAP capped AuNPs are etched by the DA molecules due to the strong affinity between DA and AuNPs, thus causing a blue shift in the absorption spectrum. The concentration of DA is quantitatively monitored by using a UV-Vis spectrometer with a limit of detection (LOD) as low as 5 nM. In addition, the results also show that the methods developed appear to have no significant problems in detecting DA in the sample even with the presence of (10 mM) common interferents such as ascorbic acid (AA), homovanillic acid (HVA), catechol (CA) and glutathione (GSH). The developed AuNP etching protocol for dopamine detection provides a novel and versatile approach for rapid biosensing applications. PMID:23016153

  14. Colorimetric growth assay for epidermal cell cultures by their crystal violet binding capacity.

    PubMed

    Bonnekoh, B; Wevers, A; Jugert, F; Merk, H; Mahrle, G

    1989-01-01

    The application of a simple, rapid, and inexpensive colorimetric growth assay was tested for human epidermal cells subcultured in uncoated plastic dishes. Cell layers were incubated with a crystal violet (CV) solution (0.2% with ethanol 2% in 0.5 M Tris-Cl buffer, pH 7.8) for 10 min at room temperature. After rinsing with 0.5 M Tris-Cl (pH 7.8) the cell layer was dried and decolorized with a sodium-dodecylsulfate solution (0.5% with ethanol 50% in 0.5 M Tris-Cl, pH 7.8) for 60 min at 37 degrees C. The extinction of the supernatant was read at the absorption maximum of 586 nm. The protein content of attached cells as classical parameter for quantifying cell growth was strongly related to CV extinction with a correlation coefficient of r = 0.98. Furthermore, the subcellular protein binding qualities of CV were analyzed. The water-soluble protein fraction of cultured epidermal cells was separated by sodium-dodecylsulfate polyacrylamide gel electrophoresis and stained with CV. We found a staining pattern which was qualitatively very similar to that of Coomassie blue, however less intense. Keratin electrophoresis revealed an affinity of CV to the 48, 50, and 56 kD cytokeratins. In conclusion, this CV assay is a reliable and simple method for the monitoring of epidermal cell growth in cultures. PMID:2482013

  15. Gold nanoparticles mediated colorimetric assay for HIV-Tat protein detection

    NASA Astrophysics Data System (ADS)

    Hashwan, Saeed S. Ba; Ruslinda, A. Rahim; Fatin, M. F.; Gopinath, Subash C. B.; Thivina, V.; Tony, V. C. S.; Arshad, M. K. Md.; Hashim, U.

    2016-07-01

    Gold-nanoparticle (AuNP) based colorimetric assays have been formulated for different biomolecular interactions. With this assay the probe such as antibody immobilized on the Au surface and in the presence of appropriate binding partner (antigen), will interact with each other on the Au surface. By following this strategy, herein we formulated a detection system with two anti-HIV-Tat antibodies, Mono (McAb) - and polyclonal (PcAb) by immobilizing them independently with different AuNPs. Under this condition, these two antibodies are under dispersed condition, and in the presence of HIV-Tat antigen, these molecules will be connected and forms the aggregation of AuNPs. This strategy yield rapid results, can be monitored by the spectral changes in UV-Vis spectrophotometry. Experiments were performed with two different methods using two anti-HIV-Tats monoclonal and one Polyclonal antibody against the antigen HIV-Tat. Between these methods conjugation of HIV-Tat and McAb on the AuNP followed by addition of PcAb yielded better results.

  16. Leaching-resistant carrageenan-based colorimetric oxygen indicator films for intelligent food packaging.

    PubMed

    Vu, Chau Hai Thai; Won, Keehoon

    2014-07-23

    Visual oxygen indicators can give information on the quality and safety of packaged food in an economic and simple manner by changing color based on the amount of oxygen in the packaging, which is related to food spoilage. In particular, ultraviolet (UV)-activated oxygen indicators have the advantages of in-pack activation and irreversibility; however, these dye-based oxygen indicator films suffer from dye leaching upon contact with water. In this work, we introduce carrageenans, which are natural sulfated polysaccharides, to develop UV-activated colorimetric oxygen indicator films that are resistant to dye leakage. Carrageenan-based indicator films were fabricated using redox dyes [methylene blue (MB), azure A, and thionine], a sacrificial electron donor (glycerol), an UV-absorbing photocatalyst (TiO2), and an encapsulation polymer (carrageenan). They showed even lower dye leakage in water than conventional oxygen indicator films, owing to the electrostatic interaction of anionic carrageenan with cationic dyes. The MB/TiO2/glycerol/carrageenan oxygen indicator film was successfully bleached upon UV irradiation, and it regained color very rapidly in the presence of oxygen compared to the other waterproof oxygen indicator films. PMID:24979322

  17. Prediction of warmed-over flavour development in cooked chicken by colorimetric sensor array.

    PubMed

    Kim, Su-Yeon; Li, Jinglei; Lim, Na-Ri; Kang, Bo-Sik; Park, Hyun-Jin

    2016-11-15

    The aim of this study was to develop a simple and rapid method based on colorimetric sensor array (CSA) for evaluation of warmed-over flavour (WOF) in cooked chicken. All samples were classified according to storage time by CSA coupled with principle component analysis (PCA) or hierarchical cluster analysis (HCA). The CSA data were used to establish prediction models with thiobarbituric acid reactive substances (TBARS), pentanal, hexanal, or heptanal associated with WOF by partial least square regression (PLSR). For the TBARS model, the coefficient of determination (rp(2)) was 0.9997 in the prediction range of 0.28-0.69mg/kg. In each of the models for pentanal, hexanal, and heptanal, all rp(2) were higher than 0.960 in the range of 0.58-2.10mg/kg, 5.50-11.69mg/kg, and 0.09-0.16mg/kg, respectively. These results demonstrate that the CSA was able to predict WOF development and to distinguish between each storage time. PMID:27283653

  18. Magnetic Bead-Based Colorimetric Immunoassay for Aflatoxin B1 Using Gold Nanoparticles

    PubMed Central

    Wang, Xu; Niessner, Reinhard; Knopp, Dietmar

    2014-01-01

    A competitive colorimetric immunoassay for the detection of aflatoxin B1 (AFB) has been established using biofunctionalized magnetic beads (MBs) and gold nanoparticles (GNPs). Aflatoxin B1-bovine serum albumin conjugates (AFB-BSA) modified MBs were employed as capture probe, which could specifically bind with GNP-labeled anti-AFB antibodies through immunoreaction, while such specific binding was competitively inhibited by the addition of AFB. After magnetic separation, the supernatant solution containing unbound GNPs was directly tested by UV-Vis spectroscopy. The absorption intensity was directly proportional to the AFB concentration. The influence of GNP size, incubation time and pH was investigated in detail. After optimization, the developed method could detect AFB in a linear range from 20 to 800 ng/L, with the limit of detection at 12 ng/L. The recoveries for spiked maize samples ranged from 92.8% to 122.0%. The proposed immunoassay provides a promising approach for simple, rapid, specific and cost-effective detection of toxins in the field of food safety. PMID:25405511

  19. Colorimetric microdetermination of captopril in pure form and in pharmaceutical formulations

    NASA Astrophysics Data System (ADS)

    Shama, Sayed Ahmed; El-Sayed Amin, Alla; Omara, Hany

    2006-11-01

    A simple, rapid, accurate, precise and sensitive colorimetric method for the determination of captopril (CAP) in bulk sample and in dosage forms is described. The method is based on oxidation of the drug by potassium permanganate in acidic medium and determination of the unreacted oxidant by measuring the decrease in absorbance for five different dyes; methylene blue (MB); acid blue 74 (AB), acid red 73 (AR), amaranth dye (AM) and acid orange 7 (AO) at a suitable λmax (660, 610, 510, 520, and 485 nm), respectively. Regression analysis of Beer's plots showed good correlation in the concentration ranges (0.4 12.5, 0.3 10, 0.5 11, 0.4 8.3 and 0.5 9.3 μg ml-1), respectively. The apparent molar absorbtivity, Sandell sensitivity, detection and quantitation limits were calculated. For more accurate results, Ringbom optimum concentration ranges were 0.5 12, 0.5 9.6, 0.6 10.5, 0.5 8.0 and 0.7 9.0 μg ml-1, respectively. The validity of the proposed method was tested by analyzing in pure and dosage forms containing CAP whether alone or in combination with hydrochlorothiazide. Statistical analysis of the results reflects that the proposed procedures are precise, accurate and easily applicable for the determination of CAP in pure form and in pharmaceutical preparations. Also, the stability constant was determined and the free energy change was calculated potentiometrically.

  20. Homogeneous detection of unamplified genomic DNA sequences based on colorimetric scatter of gold nanoparticle probes

    PubMed Central

    Storhoff, James J; Lucas, Adam D; Garimella, Viswanadham; Bao, Y Paul; Müller, Uwe R

    2005-01-01

    Nucleic acid diagnostics is dominated by fluorescence-based assays that use complex and expensive enzyme-based target or signal-amplification procedures1–6. Many clinical diagnostic applications will require simpler, inexpensive assays that can be done in a screening mode. We have developed a ‘spot-and-read’ colorimetric detection method for identifying nucleic acid sequences based on the distance-dependent optical properties of gold nanoparticles. In this assay, nucleic acid targets are recognized by DNA-modified gold probes, which undergo a color change that is visually detectable when the solutions are spotted onto an illuminated glass waveguide. This scatter-based method enables detection of zeptomole quantities of nucleic acid targets without target or signal amplification when coupled to an improved hybridization method that facilitates probe-target binding in a homogeneous format. In comparison to a previously reported absorbance-based method7, this method increases detection sensitivity by over four orders of magnitude. We have applied this method to the rapid detection of mecA in methicillin-resistant Staphylococcus aureus genomic DNA samples. PMID:15170215

  1. Quantification of ecotoxicological tests based on bioluminescence using Polaroid film.

    PubMed

    Tamminen, Manu V; Virta, Marko P J

    2007-01-01

    Assays based on the measurement of bacterial luminescence are widely used in ecotoxicology. Bacterial strains responding either to general toxicity or specific pollutants are rapid, cost-effective and easy to use. However, quantification of the signal requires relatively expensive instrumentation. We show here that the detection of luminescence of BioTox, a Vibrio fischeri-based toxicity test, and of a specific recombinant bacterial strain for arsenic determination, is possible using common Polaroid film. The exposed films can be used for visual or computer-assisted quantification of the signal. Qualitative visual comparison to standards can be used in the rapid and relatively accurate estimation of toxicity or pollutant concentration. The computer-assisted method significantly improves the accuracy and quantification of the results. The results obtained by computer-assisted quantification were in good agreement with the values obtained with a luminometer. PMID:16949132

  2. The catalytic activity of Ag2S-montmorillonites as peroxidase mimetic toward colorimetric detection of H2O2.

    PubMed

    Liu, Qingyun; Jiang, Yanling; Zhang, Leyou; Zhou, Xinpei; Lv, Xintian; Ding, Yanyuan; Sun, Lifang; Chen, Pengpeng; Yin, Hailiang

    2016-08-01

    Nanocomposites based on silver sulfide (Ag2S) and Ca-montmorillonite (Ca(2+)-MMT) were synthesized by a simple hydrothermal method. The nanocomposites were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM) and Fourier transform infrared spectra (FTIR). The as-prepared Ag2S-MMT nanocomposites were firstly demonstrated to possess intrinsic peroxidase-like activity and could rapidly catalytically oxidize the substrate 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2 to produce a blue product which can be seen by the naked eye in only one minute. The experimental results revealed that the Ag2S-MMT nanocomposites exhibit higher thermal durance. Based on the TMB-H2O2 catalyzed color reaction, the Ag2S-MMT nanocomposites were exploited as a new type of biosensor for detection and estimation of H2O2 through a simple, cheap and selective colorimetric method. PMID:27157733

  3. A low cost, safe, disposable, rapid and self-sustainable paper-based platform for diagnostic testing: lab-on-paper.

    PubMed

    Costa, M N; Veigas, B; Jacob, J M; Santos, D S; Gomes, J; Baptista, P V; Martins, R; Inácio, J; Fortunato, E

    2014-03-01

    There is a strong interest in the use of biopolymers in the electronic and biomedical industries, mainly towards low-cost applications. The possibility of developing entirely new kinds of products based on cellulose is of current interest, in order to enhance and to add new functionalities to conventional paper-based products. We present our results towards the development of paper-based microfluidics for molecular diagnostic testing. Paper properties were evaluated and compared to nitrocellulose, the most commonly used material in lateral flow and other rapid tests. Focusing on the use of paper as a substrate for microfluidic applications, through an eco-friendly wax-printing technology, we present three main and distinct colorimetric approaches: (i) enzymatic reactions (glucose detection); (ii) immunoassays (antibodies anti-Leishmania detection); (iii) nucleic acid sequence identification (Mycobacterium tuberculosis complex detection). Colorimetric glucose quantification was achieved through enzymatic reactions performed within specific zones of the paper-based device. The colouration achieved increased with growing glucose concentration and was highly homogeneous, covering all the surface of the paper reaction zones in a 3D sensor format. These devices showed a major advantage when compared to the 2D lateral flow glucose sensors, where some carryover of the coloured products usually occurs. The detection of anti-Leishmania antibodies in canine sera was conceptually achieved using a paper-based 96-well enzyme-linked immunosorbent assay format. However, optimization is still needed for this test, regarding the efficiency of the immobilization of antigens on the cellulose fibres. The detection of Mycobacterium tuberculosis nucleic acids integrated with a non-cross-linking gold nanoprobe detection scheme was also achieved in a wax-printed 384-well paper-based microplate, by the hybridization with a species-specific probe. The obtained results with the above

  4. A low cost, safe, disposable, rapid and self-sustainable paper-based platform for diagnostic testing: lab-on-paper

    NASA Astrophysics Data System (ADS)

    Costa, M. N.; Veigas, B.; Jacob, J. M.; Santos, D. S.; Gomes, J.; Baptista, P. V.; Martins, R.; Inácio, J.; Fortunato, E.

    2014-03-01

    There is a strong interest in the use of biopolymers in the electronic and biomedical industries, mainly towards low-cost applications. The possibility of developing entirely new kinds of products based on cellulose is of current interest, in order to enhance and to add new functionalities to conventional paper-based products. We present our results towards the development of paper-based microfluidics for molecular diagnostic testing. Paper properties were evaluated and compared to nitrocellulose, the most commonly used material in lateral flow and other rapid tests. Focusing on the use of paper as a substrate for microfluidic applications, through an eco-friendly wax-printing technology, we present three main and distinct colorimetric approaches: (i) enzymatic reactions (glucose detection); (ii) immunoassays (antibodies anti-Leishmania detection); (iii) nucleic acid sequence identification (Mycobacterium tuberculosis complex detection). Colorimetric glucose quantification was achieved through enzymatic reactions performed within specific zones of the paper-based device. The colouration achieved increased with growing glucose concentration and was highly homogeneous, covering all the surface of the paper reaction zones in a 3D sensor format. These devices showed a major advantage when compared to the 2D lateral flow glucose sensors, where some carryover of the coloured products usually occurs. The detection of anti-Leishmania antibodies in canine sera was conceptually achieved using a paper-based 96-well enzyme-linked immunosorbent assay format. However, optimization is still needed for this test, regarding the efficiency of the immobilization of antigens on the cellulose fibres. The detection of Mycobacterium tuberculosis nucleic acids integrated with a non-cross-linking gold nanoprobe detection scheme was also achieved in a wax-printed 384-well paper-based microplate, by the hybridization with a species-specific probe. The obtained results with the above

  5. A novel colorimetric assay of β-D-glucans in basidiomycete strains by alcian blue dye in a 96-well microtiter plate.

    PubMed

    C Semedo, Magda; Karmali, Amin; Fonseca, Luís

    2015-01-01

    Basidiomycete strains synthesize several types of β-d-glucans, which play a major role in the medicinal properties of mushrooms. Therefore, the specific quantification of these β-d-glucans in mushroom strains is of great biochemical importance. Because published assay methods for these β-d-glucans present some disadvantages, a novel colorimetric assay method for β-d-glucan with alcian blue dye was developed. The complex formation was detected by following the decrease in absorbance in the range of 620 nm and by hypsochromic shift from 620 to 606 nm (∼14 nm) in UV-Vis spectrophotometer. Analysis of variance was used for optimization of the slope of the calibration curve by using the assay mixture containing 0.017% (w/v) alcian blue in 2% (v/v) acetic acid at pH 3.0. The high-throughput colorimetric assay method on microtiter plates was used for quantification of β-d-glucans in the range of 0-0.8 μg, with a slope of 44.15 × 10(-2) and a limit of detection of 0.017 μg/well. Recovery experiments were carried out by using a sample of Hericium erinaceus, which exhibited a recovery of 95.8% for β-1,3-d-glucan. The present assay method exhibited a 10-fold higher sensitivity and a 59-fold lower limit of detection compared with the published method with congo red. β-d-glucans of several mushrooms strains were isolated from fruiting bodies and mycelia, and they were quantified by this assay method. This assay method is fast, specific, simple, and it can be used to quantify β-d-glucans from other biological sources. PMID:26317582

  6. A colorimetric sensor based on anodized aluminum oxide (AAO) substrate for the detection of nitroaromatics.

    SciTech Connect

    Liu, Y.; Wang, H. H.; Indacochea, J. E.; Wang, M. L.

    2011-12-15

    Simple and low cost colorimetric sensors for explosives detection were explored and developed. Anodized aluminum oxide (AAO) with large surface area through its porous structure and light background color was utilized as the substrate for colorimetric sensors. Fabricated thin AAO films with thickness less than {approx} 500 nm allowed us to observe interference colors which were used as the background color for colorimetric detection. AAO thin films with various thickness and pore-to-pore distance were prepared through anodizing aluminum foils at different voltages and times in dilute sulfuric acid. Various interference colors were observed on these samples due to their difference in structures. Accordingly, suitable anodization conditions that produce AAO samples with desired light background colors for optical applications were obtained. Thin film interference model was applied to analyze the UV-vis reflectance spectra and to estimate the thickness of the AAO membranes. We found that the thickness of produced AAO films increased linearly with anodization time in sulfuric acid. In addition, the growth rate was higher for AAO anodized using higher voltages. The thin film interference formulism was further validated with a well established layer by layer deposition technique. Coating poly(styrene sulfonate) sodium salt (PSS) and poly(allylamine hydrochloride) (PAH) layer by layer on AAO thin film consistently shifted its surface color toward red due to the increase in thickness. The red shift of UV-vis reflectance was correlated quantitatively to the number of layers been assembled. This sensitive red shift due to molecular attachment (increase in thickness) on AAO substrate was applied toward nitroaromatics detection. Aminopropyltrimethoxysilane (APTS) which can be attached onto AAO nanowells covalently through silanization and attract TNT molecules was coated and applied for TNT detection. UV-vis spectra of AAO with APTS shifted to the longer wavelength side due to

  7. Polyacrylic acid-coated cerium oxide nanoparticles: An oxidase mimic applied for colorimetric assay to organophosphorus pesticides.

    PubMed

    Zhang, Shi-Xiang; Xue, Shi-Fan; Deng, Jingjing; Zhang, Min; Shi, Guoyue; Zhou, Tianshu

    2016-11-15

    It is important and urgent to develop reliable and highly sensitive methods that can provide on-site and rapid detection of extensively used organophosphorus pesticides (OPs) for their neurotoxicity. In this study, we developed a novel colorimetric assay for the detection of OPs based on polyacrylic acid-coated cerium oxide nanoparticles (PAA-CeO2) as an oxidase mimic and OPs as inhibitors to suppress the activity of acetylcholinesterase (AChE). Firstly, highly dispersed PAA-CeO2 was prepared in aqueous solution, which could catalyze the oxidation of TMB to produce a color reaction from colorless to blue. And the enzyme of AChE was used to catalyze the substrate of acetylthiocholine (ATCh) to produce thiocholine (TCh). As a thiol-containing compound with reducibility, TCh can decrease the oxidation of TMB catalyzed by PAA-CeO2. Upon incubated with OPs, the enzymatic activity of AChE was inhibited to produce less TCh, resulting in more TMB catalytically oxidized by PAA-CeO2 to show an increasing blue color. The two representative OPs, dichlorvos and methyl-paraoxon, were tested using our proposed assay. The novel assay showed notable color change in a concentration-dependent manner, and as low as 8.62 ppb dichlorvos and 26.73 ppb methyl-paraoxon can be readily detected. Therefore, taking advantage of such oxidase-like activity of PAA-CeO2, our proposed colorimetric assay can potentially be a screening tool for the precise and rapid evaluation of the neurotoxicity of a wealth of OPs. PMID:27208478

  8. Colorimetric solid-phase extraction method for Cu(II) ion determination using 2-hydroxybenzaldehyde benzoylhydrazone as sensing reagent.

    PubMed

    Espada-Bellido, Estrella; Galindo-Riaño, Maria Dolores; García-Vargas, Manuel; Narayanaswamy, Ramaier

    2014-01-01

    A new sensor based on the use of 2-hydroxybenzaldehyde benzoylhydrazone as a colorimetric reagent immobilized onto styrenedivinylbenzene disks has been carried out for the determination of Cu(II) ions within several minutes. The sensor is designed on a rapid and easy two-step procedure: (1) the extraction of Cu(II) ions onto a disk loaded with the copper-selective colorimetric reagent and (2) the determination of the complexed analyte directly on the surface of the disk using diffuse reflectance measurements at 400 nm. The color of the disk changed from white to green in the presence of Cu(II) ions. The work herein details the optimization of the sensing system employing a fractional factorial design 3(3-1) considering three variables (pH, immobilization time, and amount of ligand immobilized onto the disk). The Pareto chart and response surfaces in a spherical domain indicated that the optimum conditions for the sensing of copper ions were pH = 7, with a ligand immobilization time of 10 min and 6.25 mg of reagent loaded onto the disk. Under the optimum conditions, the analytical parameters of the proposed method were determined. The calibration graph was linear over the range of 0 to 2.5 mg L(-1) of Cu(II) with a detection limit of 0.21 mg L(-1). The relative standard deviation for six measurements of 1 mg L(-1) of Cu(II) was found to be 4.87%. The interference from inorganic salts and other metals was found not to be of major concern when monitoring copper ions in water samples. The simplicity and rapidity of this technique make it convenient and amenable for on-site and routine analysis. PMID:24694697

  9. Operator-Friendly Technique and Quality Control Considerations for Indigo Colorimetric Measurement of Ozone Residual

    EPA Science Inventory

    Drinking water ozone disinfection systems measure ozone residual concentration, C, for regulatory compliance reporting of concentration-times-time (CT), and the resultant log-inactivation of virus, Giardia and Cryptosporidium. The indigotrisulfonate (ITS) colorimetric procedure i...

  10. A novel highly selective colorimetric sensor for aluminum (III) ion using Schiff base derivative.

    PubMed

    Wang, Dong-Fang; Ke, Ying-Chang; Guo, Hong-Xu; Chen, Jianhua; Weng, Wen

    2014-03-25

    A novel colorimetric sensor, 2-hydroxy naphthaldehyde isonicotinoyl hydrazone (HINH), was easily synthesized by the condensation of isoniazid and 2-hydroxy-1-naphthaldehyde. The as-prepared compound showed effective colorimetric single selectivity and high sensitivity for aluminum cation in CH3CN/H2O (1:3) binary solutions. The detection limit is 1.0×10(-8) M Al(3+) based on UV-vis changes. PMID:24316541

  11. Quantitative analysis of modeled ATP hydrolysis in water by a colorimetric sensor array.

    PubMed

    Minami, Tsuyoshi; Emami, Fereshteh; Nishiyabu, Ryuhei; Kubo, Yuji; Anzenbacher, Pavel

    2016-06-14

    Self-assembled colorimetric sensors have been prepared from Zn(II)-DPA-attached phenylboronic acid (·Zn) and catechol-type dyes. The ·Zn-dye sensors display selectivity towards oligophosphate over monophosphates. The colorimetric sensor assay (·Zn-dye) is utilized to monitor a model of a metabolic reaction where ATP is hydrolyzed to pyrophosphate (PPi) and AMP. PMID:27241171

  12. Colorimetric visualization of glucose at the submicromole level in serum by a homogenous silver nanoprism-glucose oxidase system.

    PubMed

    Xia, Yunsheng; Ye, Jingjing; Tan, Kanghui; Wang, Jiajing; Yang, Guang

    2013-07-01

    In this study, we design a homogeneous system consisting of Ag nanoprisms and glucose oxidase (GOx) for simple, sensitive, and low-cost colorimetric sensing of glucose in serum. The unmodified Ag nanoprisms and GOx are first mixed with each other. Glucose is then added in the homogeneous mixture. Finally, the nanoplates are etched from triangle to round by H2O2 produced by the enzymatic oxidation, which leads to a more than 120 nm blue shift of the surface plasmon resonance (SPR) absorption band of the Ag nanoplates. This large wavelength shift can be used not only for visual detection (from blue to mauve) of glucose by naked eyes but for reliable and convenient glucose quantification in the range from 2.0 × 10(-7) to 1.0 × 10(-4) M. The detection limit is as low as 2.0 × 10(-7) M, because the used Ag nanoprisms possess (1) highly reactive edges/tips and (2) strongly tip sharpness and aspect ratio dependent SPR absorption. Owing to ultrahigh sensitivity, only 10-20 μL of serum is enough for a one-time determination. The proposed glucose sensor has great potential in the applications of point-of-care diagnostics, especially for third-world countries where high-tech diagnostics aids are inaccessible to the bulk of the population. PMID:23706061

  13. A Colorimetric Interdental Probe as a Standard Method to Evaluate Interdental Efficiency of Interdental Brush

    PubMed Central

    Bourgeois, D.; Carrouel, F.; Llodra, J.C.; Bravo, M.; Viennot, S.

    2015-01-01

    The aim of this study is to evaluate the concordance between the empirical choice of interdental brushes of different diameters compared to the gold standard, the IAP CURAPROX© calibrating colorimetric probe. It is carried out with the aim of facilitating the consensus development of best practices. All the subjects’ interproximal spaces were evaluated using the reference technique (colorimetric probe), then after a time lapse of 1.2 ± 0.2 hours, using the empirical clinical technique (brushes) by the same examiner. Each examiner explored 3 subjects. The order the patients were examined with the colorimetric interdental probe (CIP) was random. 446 sites were selected in the study out of 468 potential sites. The correspondence of scores between interdental bushes vs. colorimetric probe is 43.0% [95%-CI: 38.5-47.6]. In 33.41% of the 446 sites, the brush is inferior to the probe; in 23.54% of cases, the brush is superior to the probe. Among the discrepancies there is thus a tendency for the subjects to use brushes with smaller diameter than that recommended by the colorimetric probe. This review has found very high-quality evidence that colorimetric probes plus interdental brushing is more beneficial than interdental brushing alone for increase the concordance between the empirical choice of interdental brushes of different diameters compared to the gold standard. Uncertainties remain and further research is required to provide detailed data on user satisfaction. PMID:26966470

  14. Development and validation of a high-throughput micro solid-phase extraction method coupled with ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry for rapid identification and quantification of phenolic metabolites in human plasma and urine.

    PubMed

    Feliciano, Rodrigo P; Mecha, Elsa; Bronze, Maria R; Rodriguez-Mateos, Ana

    2016-09-16

    A rapid and high-throughput micro-solid phase extraction (μ-SPE) method coupled with ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC Q-TOF MS) analysis was optimized and validated for the quantification of 67 (poly)phenol metabolites in human plasma and urine using authentic standards. The method was fully validated in terms of specificity, linearity, method detection limit (MDL), method quantification limit (MQL), repeatability, intra- and inter-day precision, accuracy and matrix effects. The method proved to be specific and results showed linearity of responses for all compounds, with MDL ranging between 0.04nM and 86nM in plasma and between 0.01nM and 136nM in urine. MQL ranged between 0.14nM and 286nM in plasma and between 0.03nM and 465nM in urine. Repeatability varied between 1.7 and 9.2% in plasma and between 2.2% and 10.4% in urine. Median precision values of 8.7 and 11.5% (intra-day), and 10.8% and 10.0% (inter-day) were obtained in plasma and urine, respectively. The median recovery was 89% in both biological matrices. Matrix effects were determined and median values of -1.2% and -6.8% in plasma and urine were obtained. After method validation, 49 and 57 compounds, including phase II and gut microbial metabolites, were quantified in plasma and urine, respectively, following cranberry juice consumption. This methodology can be applied to large-scale human dietary intervention trials allowing for high sample throughput. PMID:27527878

  15. Quantification of nonclassicality

    NASA Astrophysics Data System (ADS)

    Gehrke, C.; Sperling, J.; Vogel, W.

    2012-11-01

    To quantify single-mode nonclassicality, we start from an operational approach. A positive semidefinite observable is introduced to describe a measurement setup. The quantification is based on the negativity of the normally ordered version of this observable. Perfect operational quantumness corresponds to the quantum-noise-free measurement of the chosen observable. Surprisingly, even moderately squeezed states may exhibit perfect quantumness for a properly designed measurement. The quantification is also considered from an axiomatic viewpoint, based on the algebraic structure of the quantum states and the quantum superposition principle. Basic conclusions from both approaches are consistent with this fundamental principle of the quantum world.

  16. Specific S-nitrosothiol (thionitrite) quantification as solution nitrite after vanadium(III) reduction and ozone-chemiluminescent detection.

    PubMed

    Ewing, J F; Janero, D R

    1998-09-01

    Increasing evidence suggests that S-nitrosothiols (thionitrites) might represent naturally occurring nitric oxide surrogates and function as intermediates in nitrogen monoxide metabolism. A facile, sensitive, and selective micromethod has been developed and validated for quantification of S-nitrosothiols as their mercury-displaceable nitrogen monoxide content. In this method, brief (5-min), room-temperature pretreatment of S-nitrosothiol with a molar excess of aqueous mercuric chloride was used to liberate into solution, quantitatively, the nitrogen monoxide moiety, which rapidly and quantitatively converted to its stable solution end-product, nitrite. Solution nitrite was reduced back to nitric oxide with vanadium(III), and the nitric oxide was detected by gas-phase chemiluminescence after reaction with ozone in a commercial nitric oxide analyzer. A linear relationship was observed between S-nitrosothiol-bound nitrogen monoxide and ozone-chemiluminescent detector response over a wide range (16.3-3500 pmol) of nitric oxide, as generated by reaction of vanadium(III) with either nitrite standard or mercury-treated S-nitrosothiol. Assay response was quantitatively identical for equivalent amounts of nitrite and S-nitrosothiol-bound nitrogen monoxide. The method displayed 96% selectivity for nitrite vs. nitrate and negligible (<2%) interference by nitrosated compounds bearing nitrogen monoxide moieties bound to either nitrogen or carbon. The lower limits of quantitative sensitivity and qualitative detection were below 50 and 20 pmol S-nitrosothiol-bound nitrogen monoxide-equivalents, respectively. The intraday and interday coefficients of variation did not exceed 8%. This technique has been applied to quantify structurally diverse natural and synthetic S-nitrosothiols with quantitative recovery from complex biological samples such as culture media and plasma at levels of nitrogen monoxide-equivalents undetectable by the popular Saville colorimetric method. PMID:9741600

  17. Colour thresholding and objective quantification in bioimaging

    NASA Technical Reports Server (NTRS)

    Fermin, C. D.; Gerber, M. A.; Torre-Bueno, J. R.

    1992-01-01

    Computer imaging is rapidly becoming an indispensable tool for the quantification of variables in research and medicine. Whilst its use in medicine has largely been limited to qualitative observations, imaging in applied basic sciences, medical research and biotechnology demands objective quantification of the variables in question. In black and white densitometry (0-256 levels of intensity) the separation of subtle differences between closely related hues from stains is sometimes very difficult. True-colour and real-time video microscopy analysis offer choices not previously available with monochrome systems. In this paper we demonstrate the usefulness of colour thresholding, which has so far proven indispensable for proper objective quantification of the products of histochemical reactions and/or subtle differences in tissue and cells. In addition, we provide interested, but untrained readers with basic information that may assist decisions regarding the most suitable set-up for a project under consideration. Data from projects in progress at Tulane are shown to illustrate the advantage of colour thresholding over monochrome densitometry and for objective quantification of subtle colour differences between experimental and control samples.

  18. Correlating human color similarity judgments and colorimetric representations

    NASA Astrophysics Data System (ADS)

    Vertan, Constantin C.; Ciuc, Mihai; Stoica, A.; Zamfir, Marta; Buzuloiu, Vasile V.; Fernandez-Maloigne, Christine

    2003-10-01

    A color similarity test was conducted on the 24 color patches of a Gretag Macbeth color checker. Color similarities were measured either by distances between standard colorimetric representations (such as RGB, Lab or spectral reflectance curves) or by human observer judgments. In each case, the dissimilarity matrix was processed by a classical, metric, multidimensional scaling algorithm, in order to produce a visually-interpretable two-dimensional plot of color dissimilarity. The analysis of the plots produces some interesting conclusions. First, the plots produced by the Lab, RGB and spectral representations exhibit very evident variation axes according to the luminance and basic chromatic differences (red-green, blue-yellow). This behavior (trivial for the Lab representation) suggests that the color similarity measurement by chromatic differences is implicitly embedded in the RGB and spectral representations. The color dissimilarity plots associated to the human judgments (for any individual, as well as for an "average" observer) exhibit a different organization, which mixes hue, saturation and luminance (HSV). According to these plots, the human similarity judgment is not entirely HSV-based. We prove that it is possible to obtain the same color dissimilarity plots if a fuzzy color model is assumed. The fuzzy color model provides similarity coefficients (similarity degrees) between pairs of colors, based on their inter-distance, according to an imposed "color confusion" control parameter, which seems to be relevant for the human vision.

  19. Colorimetric assay of blood coagulation factor XIII in plasma.

    PubMed

    Lee, K N; Birckbichler, P J; Patterson, M K

    1988-05-01

    In this new colorimetric assay for Factor XIII in plasma, 5-(biotinamido)pentylamine is used as the amine substrate. Factor XIII, a zymogen, is transformed by thrombin and Ca2+ to active Factor XIIIa, and the incorporation of 5-(biotinamido)pentylamine into N,N-dimethylcasein is used to measure catalytically active Factor XIIIa. The biotinylated enzymatic product is immobilized onto 96-well microtiter plates, complexed with streptavidin-beta-galactosidase, and the absorbance at 405 nm is monitored for production of p-nitrophenol from p-nitrophenyl-beta-D-galactopyranoside. Concentrations of N,N-dimethylcasein, 5-(biotinamido)pentylamine, Ca2+, and thrombin were chosen to allow near-maximum velocity of amine incorporation. A linear relationship was obtained between assay product and plasma volume, from 0.5 to 50 microL of plasma. Results correlated well (r greater than 0.924) with those from the most frequently utilized radiometric filter-paper assay for Factor XIII. The method appears to be ideal for routine diagnostic estimation of Factor XIII in plasma because of its simplicity, its lack of use of radioisotopes, and its potential for assay of large numbers of samples by use of microtiter plates and automated plate readers. PMID:2897256

  20. Direct colorimetric method for determination of organophosphates in human urine.

    PubMed

    Namera, A; Utsumi, Y; Yashiki, M; Ohtani, M; Imamura, T; Kojima, T

    2000-01-20

    A simple and sensitive method for determination of organophosphorus pesticides in human urine was developed by detecting the color complexes which resulted from reactions of organophosphorus pesticides and 4-(4-nitrobenzyl)pyridine (NBP) in urine. Based on studies of reaction conditions, e.g. reaction temperature and time, and reagent concentration, a colorimetric method was established. A 0.1-ml volume of NBP (45% in acetone) was added to a 1.0-ml volume of a urine sample, and the mixture was heated at 100 degrees C for 20 min. After cooling, 0.1 ml of tetraethylenepentamine was added. The organophosphorus pesticides showed a characteristic purplish blue color and the coloring complexes which were produced were stable for several hours. Furthermore, these complexes could be determined spectrophotometrically. The detection limits were 0.10-10 microg/ml in urine. The required time for analysis was approximately 30 min for one sample. Comparing the result of the proposed method with those of the GC-MS method, the results were similar for the 12 poisoning cases studied. Thus, the proposed method is useful for detection of these pesticides in critical care practices. PMID:10612713

  1. Colorimetric multiplexed immunoassay for sequential detection of tumor markers.

    PubMed

    Wang, Jing; Cao, Ya; Xu, Yuanyuan; Li, Genxi

    2009-10-15

    In this paper, a very simple and easily-operated colorimetric multiplexed immunoassay method for sequential detection of tumor markers has been presented. Magnetic microparticles which are conjugated with biotinylated antibodies are firstly added into the test solution. After fast magnetic collection, these complexes are separated from non-specific proteins. Through different enzymatic reactions of 3,3',5,5'-tetramethylbenzidine (TMB) and o-phenylenediamine (OPD) catalyzed by horseradish peroxidase molecules which are loaded on the surfaces of gold nanoparticles, two antigens carcinoembryonic antigen and alpha-fetoprotein can be detected even with naked eyes. The detection limit obtained from the spectrophotometric measurements is as low as 0.02 ng/mL. This proposed method also has high specificity and reproducibility, as well as excellent efficiency of 94 min for the detection of serum samples. So, this new multiplexed immunoassay method might be a promising approach for the diagnosis of cancer and some other diseases in clinical applications. PMID:19726177

  2. Colorimetric detection of catalytic reactivity of nanoparticles in complex matrices.

    PubMed

    Corredor, Charlie; Borysiak, Mark D; Wolfer, Jay; Westerhoff, Paul; Posner, Jonathan D

    2015-03-17

    There is a need for new methodologies to quickly assess the presence and reactivity of nanoparticles (NPs) in commercial, environmental, and biological samples since current detection techniques require expensive and complex analytical instrumentation. Here, we investigate a simple and portable colorimetric detection assay that assesses the surface reactivity of NPs, which can be used to detect the presence of NPs, in complex matrices (e.g., environmental waters, serum, urine, and in dissolved organic matter) at as low as part per billion (ppb) or ng/mL concentration levels. Surface redox reactivity is a key emerging property related to potential toxicity of NPs with living cells, and is used in our assays as a key surrogate for the presence of NPs and a first tier analytical strategy toward assessing NP exposures. We detect a wide range of metal (e.g., Ag and Au) and oxide (e.g., CeO2, SiO2, VO2) NPs with a diameter range of 5 to 400 nm and multiple capping agents (tannic acid (TA), polyvinylpyrrolidone (PVP), branched polyethylenimine (BPEI), polyethylene glycol (PEG)). This method is sufficiently sensitive (ppb levels) to measure concentrations typically used in toxicological studies, and uses inexpensive, commercially available reagents. PMID:25635807

  3. Point-of-care colorimetric detection with a smartphone.

    PubMed

    Shen, Li; Hagen, Joshua A; Papautsky, Ian

    2012-11-01

    Paper-based immunoassays are becoming powerful and low-cost diagnostic tools, especially in resource-limited settings. Inexpensive methods for quantifying these assays have been shown using desktop scanners, which lack portability, and cameras, which suffer from the ever changing ambient light conditions. In this work, we introduce a novel approach of quantifying colors of colorimetric diagnostic assays with a smartphone that allows high accuracy measurements in a wide range of ambient conditions, making it a truly portable system. Instead of directly using the red, green, and blue (RGB) intensities of the color images taken by a smartphone camera, we use chromaticity values to construct calibration curves of analyte concentrations. We demonstrate the high accuracy of this approach in pH measurements with linear response ranges of 1-12. These results are comparable to those reported using a desktop scanner or silicon photodetectors. To make the approach adoptable under different lighting conditions, we developed a calibration technique to compensate for measurement errors due to variability in ambient light. This technique is applicable to a number of common light sources, such as sun light, fluorescent light, or smartphone LED light. Ultimately, the entire approach can be integrated in an "app" to enable one-click reading, making our smartphone based approach operable without any professional training or complex instrumentation. PMID:22996728

  4. Color To Colorant Conversions In A Colorimetric Separation System

    NASA Astrophysics Data System (ADS)

    Holub, Richard; Kearsley, Wayne

    1990-06-01

    Within several years of establishment of the CIE's Standard Observer, Neugebauer based upon it a device-independent interpretation of ink mixture in halftone color reproduction. Despite limitations of the model, his approach is of fundamental importance at a time in which document preparation in distributed computing environments is coming of age. Fruition of this technology demands a device independent or colorimetric method for representing and rendering color. A commercial, digital system for implementing such an approach (Kodak Designmaster 8000) has been described and will be reviewed here with particular regard to the output of color imagery on four-color printers. Inversion of Neugebauer's model for conversion of color to device specific signals must be constrained by the color gamut of the output medium and the utilization of the black printer (Gray Component Replacement). The interaction of the black printer with the device gamut and its effect on image quality and metamerism will receive detailed consideration. It will be shown that colorimetry is a powerful tool in maintaining accurate color reproduction during substitution of black for chromatic components, but that properties of the colorants must also be considered in order to realize commercially acceptable GCR. Section 7 of the paper is an enumeration of summary conclusions.

  5. Quercetin as colorimetric reagent for determination of zirconium

    USGS Publications Warehouse

    Grimaldi, F.S.; White, C.E.

    1953-01-01

    Methods described in the literature for the determination of zirconium are generally designed for relatively large amounts of this element. A good procedure using colorimetric reagent for the determination of trace amounts is desirable. Quercetin has been found to yield a sensitive color reaction with zirconium suitable for the determination of from 0.1 to 50?? of zirconium dioxide. The procedure developed involves the separation of zirconium from interfering elements by precipitation with p-dimethylaminoazophenylarsonic acid prior to its estimation with quercetin. The quercetin reaction is carried out in 0.5N hydrochloric acid solution. Under the operating conditions it is indicated that quercetin forms a 2 to 1 complex with zirconium; however, a 2 to 1 and a 1 to 1 complex can coexist under special conditions. Approximate values for the equilibrium constants of the complexes are K1 = 0.33 ?? 10-5 and K2 = 1.3 ?? 10-9. Seven Bureau of Standards samples of glass sands and refractories were analyzed with excellent results. The method described should find considerable application in the analysis of minerals and other materials for macro as well as micro amounts of zirconium.

  6. Adaptive Fourier modeling for quantification of tremor.

    PubMed

    Riviere, C N; Reich, S G; Thakor, N V

    1997-06-01

    A new computational method for quantification of tremor, the weighted frequency Fourier linear combiner (WFLC), is presented. This technique rapidly determines the frequency and amplitude of tremor by adjusting its filter weights according to a gradient search method. It provides continual tracking of frequency and amplitude modulations over the course of a test. By quantifying time-varying characteristics, the WFLC assists in correctly interpreting the results of spectral analysis, particularly for recordings exhibiting multiple spectral peaks. It therefore supplements spectral analysis, providing a more accurate picture of tremor than spectral analysis alone. The method has been incorporated into a desktop tremor measurement system to provide clinically useful analysis of tremor recorded during handwriting and drawing using a digitizing tablet. Simulated data clearly demonstrate tracking of variations in frequency and amplitude. Clinical recordings then show specific examples of quantification of time-varying aspects of tremor. PMID:9210577

  7. Highly Sensitive Colorimetric Cancer Cell Detection Based on Dual Signal Amplification.

    PubMed

    Yu, Tao; Dai, Pan-Pan; Xu, Jing-Juan; Chen, Hong-Yuan

    2016-02-24

    Facile and efficient detection of cancer cells at their preclinical stages is one of the central challenges in cancer diagnostics. A direct, rapid, highly sensitive and specific biosensor for detection of cancer biomarkers is desirable in early diagnosis and prognosis of cancer. In this work, we developed, for the first time, an easy and intuitive dispersion-dominated colorimetric strategy for cancer cell detection based on combining multi-DNA released from an aptamer scaffold with cyclic enzymatic amplification, which was triggered by aptamer DNA conformational switch and demonstrated by non-cross-linking gold nanoparticles (Au NPs) aggregation. First, five kinds of messenger DNAs (mDNAs) were aligned on the cancer cell aptamers modified on magnetic beads (MBs) to form mDNAs-Apt-MBs biocompatible nanosensors. In the presence of target cells, the aptamer would bind to the receptors on the cell membranes, and mDNAs would be released, resulting in the first amplification that one biological binding event would cause the release of multiple kinds of mDNAs simultaneously. After magnetic separation, the released mDNAs were introduced into the cyclic enzymatic amplification to cleave more single strand DNA (ssDNA) fragments. Instead of modification of Au NPs, these fragments and mDNAs could be adsorbed on the surface of Au NPs to prevent particle aggregation and ensure the stability and color of solution in high salt environments. The linear response for HL-60 cells in a concentration range from 10 to 10(4) cells was obtained with a detection limit of four cells in buffer solution. Moreover, the feasibility of the proposed strategy was demonstrated in a diluted serum sample. This dual signal amplification method can be extended to other types of cancer cells, which has potential application in point-of-care cancer diagnosis. PMID:26824724

  8. A colorimetric method for the molecular weight determination of polyethylene glycol using gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Ling, Kai; Jiang, Hongyan; Zhang, Qiqing

    2013-12-01

    A gold nanoparticle (AuNP)-based colorimetric method was developed for the molecular weight (MW) determination of polyethylene glycol (PEG), a commonly used hydrophilic polymer. Addition of a salt solution to PEG-coated AuNP solutions helps in screening the electrostatic repulsion between nanoparticles and generating a color change of the solutions from wine red to blue in 10 min in accordance with the MW of PEG, which illustrates the different stability degrees (SDs) of the AuNPs. The SDs are calculated by the absorbance ratios of the stable to the aggregated AuNPs in the solution. The root mean square end-to-end length (< h 2>1/2) of PEG molecules shows a linear fit to the SDs of the PEG-coated AuNPs in a range of 1.938 ± 0.156 to 10.151 ± 0.176 nm. According to the Derjaguin-Landau-Verwey-Overbeek theory, the reason for this linear relationship is that the thickness of the PEG adlayer is roughly equivalent to the < h 2>1/2 of the PEG molecules in solution, which determines the SDs of the AuNPs. Subsequently, the MW of the PEG can be obtained from its < h 2>1/2 using a mathematical relationship between < h 2>1/2 and MW of PEG molecule. Applying this approach, we determined the < h 2>1/2 and the MW of four PEG samples according to their absorbance values from the ordinary ultraviolet-visible spectrophotometric measurements. Therefore, the MW of PEG can be distinguished straightforwardly by visual inspection and determined by spectrophotometry. This novel approach is simple, rapid, and sensitive.

  9. A Method for Selecting Structure-switching Aptamers Applied to a Colorimetric Gold Nanoparticle Assay

    PubMed Central

    Martin, Jennifer A.; Smith, Joshua E.; Warren, Mercedes; Chávez, Jorge L.; Hagen, Joshua A.; Kelley-Loughnane, Nancy

    2015-01-01

    Small molecules provide rich targets for biosensing applications due to their physiological implications as biomarkers of various aspects of human health and performance. Nucleic acid aptamers have been increasingly applied as recognition elements on biosensor platforms, but selecting aptamers toward small molecule targets requires special design considerations. This work describes modification and critical steps of a method designed to select structure-switching aptamers to small molecule targets. Binding sequences from a DNA library hybridized to complementary DNA capture probes on magnetic beads are separated from nonbinders via a target-induced change in conformation. This method is advantageous because sequences binding the support matrix (beads) will not be further amplified, and it does not require immobilization of the target molecule. However, the melting temperature of the capture probe and library is kept at or slightly above RT, such that sequences that dehybridize based on thermodynamics will also be present in the supernatant solution. This effectively limits the partitioning efficiency (ability to separate target binding sequences from nonbinders), and therefore many selection rounds will be required to remove background sequences. The reported method differs from previous structure-switching aptamer selections due to implementation of negative selection steps, simplified enrichment monitoring, and extension of the length of the capture probe following selection enrichment to provide enhanced stringency. The selected structure-switching aptamers are advantageous in a gold nanoparticle assay platform that reports the presence of a target molecule by the conformational change of the aptamer. The gold nanoparticle assay was applied because it provides a simple, rapid colorimetric readout that is beneficial in a clinical or deployed environment. Design and optimization considerations are presented for the assay as proof-of-principle work in buffer to

  10. Colorimetric sensing of copper(II) based on catalytic etching of gold nanoparticles.

    PubMed

    Liu, Ruili; Chen, Zhaopeng; Wang, Shasha; Qu, Chengli; Chen, Lingxin; Wang, Zhuo

    2013-08-15

    Based on the catalytic etching of gold nanoparticles (AuNPs), a label-free colorimetric probe was developed for the detection of Cu(2+) in aqueous solutions. AuNPs were first stabilized by hexadecyltrimethylammonium bromide in NH3-NH4Cl (0.6M/0.1M) solutions. Then thiosulfate (S2O3(2-)) ions were introduced and AuNPs were gradually dissolved by dissolved oxygen. With the further addition of Cu(2+), Cu(NH3)4(2+) oxidized AuNPs to produce Au(S2O3)2(3-) and Cu(S2O3)3(5-), while the later was oxid-ized to Cu(NH3)4(2+) again by dissolved oxygen. The dissolving rate of AuNPs was thereby remarkably promoted and Cu(2+) acted as the catalyst. The process went on due to the sufficient supply of dissolved oxygen and AuNPs were rapidly etched. Meanwhile, a visible color change from red to colorless was observed. Subsequent tests confirmed such a non-aggregation-based method as a sensitive (LOD=5.0 nM or 0.32 ppb) and selective (at least 100-fold over other metal ions except for Pb(2+) and Mn(2+)) way for the detection of Cu(2+) (linear range, 10-80 nM). Moreover, our results show that the color change induced by 40 nM Cu(2+) can be easily observed by naked eyes, which is particularly applicable to fast on-site investigations. PMID:23708534

  11. A single marker choice strategy in simultaneous characterization and quantification of multiple components by rapid resolution liquid chromatography coupled with triple quadrupole tandem mass spectrometry (RRLC-QqQ-MS).

    PubMed

    Ning, Zhangchi; Liu, Zhenli; Song, Zhiqian; Zhao, Siyu; Dong, Yunzhuo; Zeng, Honglian; Shu, Yisong; Lu, Cheng; Liu, Yuanyan; Lu, Aiping

    2016-05-30

    Single standard to determine multi-components (SSDMC) method has been accepted as an efficient technique for the quality control of Traditional Chinese medicines (TCMs), especially for overcoming the shortage of reference standards. HPLC-UV methods have been applied to establish SSDMC method for quantitative analysis in several plant medicines and Chinese patent medicines, however, no LC-MS methods have been used. The purpose of this study is to put forward an improved strategy for the choice of single marker in SSDMC using rapid resolution liquid chromatography coupled with triple quadrupole tandem mass spectrometry (RRLC-QqQ-MS). Five different Panax genus plants, recorded in the Chinese Pharmacopeia 2015 edition, were used as research subjects. An improved SSDMC strategy for simultaneous characterization and determination of 18 bioactive saponins in five Panax plants was put forward, and which was validated to be more superior. Then, it was fully investigated with respect to linearity, LODs, LOQs, precision and accuracy. Coupling with multivariate statistical analysis, the established and validated SSDMC strategy could be successively used in discrimination of the five Panax genus plants. PMID:26955755

  12. Quantification of Hepatic Steatosis With Dual-Energy Computed Tomography

    PubMed Central

    Artz, Nathan S.; Hines, Catherine D.G.; Brunner, Stephen T.; Agni, Rashmi M.; Kühn, Jens-Peter; Roldan-Alzate, Alejandro; Chen, Guang-Hong; Reeder, Scott B.

    2012-01-01

    Objective The aim of this study was to compare dual-energy computed tomography (DECT) and magnetic resonance imaging (MRI) for fat quantification using tissue triglyceride concentration and histology as references in an animal model of hepatic steatosis. Materials and Methods This animal study was approved by our institution's Research Animal Resource Center. After validation of DECT and MRI using a phantom consisting of different triglyceride concentrations, a leptin-deficient obese mouse model (ob/ob) was used for this study. Twenty mice were divided into 3 groups based on expected levels of hepatic steatosis: low (n = 6), medium (n = 7), and high (n = 7) fat. After MRI at 3 T, a DECT scan was immediately performed. The caudate lobe of the liver was harvested and analyzed for triglyceride concentration using a colorimetric assay. The left lateral lobe was also extracted for histology. Magnetic resonance imaging fat-fraction (FF) and DECT measurements (attenuation, fat density, and effective atomic number) were compared with triglycerides and histology. Results Phantom results demonstrated excellent correlation between triglyceride content and each of the MRI and DECT measurements (r2 ≥ 0.96, P ≤ 0.003). In vivo, however, excellent triglyceride correlation was observed only with attenuation (r2 = 0.89, P < 0.001) and MRI-FF (r2 = 0.92, P < 0.001). Strong correlation existed between attenuation and MRI-FF (r2 = 0.86, P < 0.001). Nonlinear correlation with histology was also excellent for attenuation and MRI-FF. Conclusions Dual-energy computed tomography (CT) data generated by the current Gemstone Spectral Imaging analysis tool do not improve the accuracy of fat quantification in the liver beyond what CT attenuation can already provide. Furthermore, MRI may provide an excellent reference standard for liver fat quantification when validating new CT or DECT methods in human subjects. PMID:22836309

  13. Quantificational logic of context

    SciTech Connect

    Buvac, Sasa

    1996-12-31

    In this paper we extend the Propositional Logic of Context, to the quantificational (predicate calculus) case. This extension is important in the declarative representation of knowledge for two reasons. Firstly, since contexts are objects in the semantics which can be denoted by terms in the language and which can be quantified over, the extension enables us to express arbitrary first-order properties of contexts. Secondly, since the extended language is no longer only propositional, we can express that an arbitrary predicate calculus formula is true in a context. The paper describes the syntax and the semantics of a quantificational language of context, gives a Hilbert style formal system, and outlines a proof of the system`s completeness.

  14. 4-mercaptophenylboronic acid functionalized gold nanoparticles for colorimetric sialic acid detection.

    PubMed

    Sankoh, Supannee; Thammakhet, Chongdee; Numnuam, Apon; Limbut, Warakorn; Kanatharana, Proespichaya; Thavarungkul, Panote

    2016-11-15

    A simple and selective colorimetric sensor for sialic acid detection, based on the aggregation of 4-mercaptophenylboronic acid functionalized gold nanoparticles (4-MPBA-AuNPs) was developed. The color of the solution changed from wine-red to blue after binding with sialic acid. The colorimetric sensor provided good analytical performances with a linear dynamic range of 80µM to 2.00mM and a 68±2µM limit of detection without any effect from possible interferences and sample matrix. In addition, the quantitative results were obtained within only 10min. This developed sensor was used to detect sialic acid in blood serum samples and the results were in good agreement with those from the current periodate-resorcinol method (P>0.05) thus indicating that this developed colorimetric sensor can be used as an alternative method for sialic acid detection with a shorter analysis time and a high accuracy. PMID:27266659

  15. Colorimetric detection of bisphenol A based on unmodified aptamer and cationic polymer aggregated gold nanoparticles.

    PubMed

    Zhang, Dongwei; Yang, Jiayi; Ye, Jing; Xu, Lurong; Xu, Hanchu; Zhan, Shenshan; Xia, Bing; Wang, Lumei

    2016-04-15

    In this study, a colorimetric method was exploited to detect bisphenol A (BPA) based on BPA-specific aptamer and cationic polymer-induced aggregation of gold nanoparticles (AuNPs). The principle of this assay is very classical. The aggregation of AuNPs was induced by the concentration of cationic polymer, which is controlled by specific recognition of aptamer with BPA and the reaction of aptamer and cationic polymer forming "duplex" structure. This method enables colorimetric detection of BPA with selectivity and a detection limit of 1.50 nM. In addition, this colorimetric method was successfully used to determine spiked BPA in tap water and river water samples. PMID:26820097

  16. Colorimetric characterization of digital cameras with unrestricted capture settings applicable for different illumination circumstances

    NASA Astrophysics Data System (ADS)

    Fang, Jingyu; Xu, Haisong; Wang, Zhehong; Wu, Xiaomin

    2016-05-01

    With colorimetric characterization, digital cameras can be used as image-based tristimulus colorimeters for color communication. In order to overcome the restriction of fixed capture settings adopted in the conventional colorimetric characterization procedures, a novel method was proposed considering capture settings. The method calculating colorimetric value of the measured image contains five main steps, including conversion from RGB values to equivalent ones of training settings through factors based on imaging system model so as to build the bridge between different settings, scaling factors involved in preparation steps for transformation mapping to avoid errors resulted from nonlinearity of polynomial mapping for different ranges of illumination levels. The experiment results indicate that the prediction error of the proposed method, which was measured by CIELAB color difference formula, reaches less than 2 CIELAB units under different illumination levels and different correlated color temperatures. This prediction accuracy for different capture settings remains the same level as the conventional method for particular lighting condition.

  17. Simple, field portable colorimetric detection device for organic peroxides and hydrogen peroxide

    DOEpatents

    Pagoria, Philip F.; Mitchell, Alexander R.; Whipple, Richard E.; Carman, M. Leslie; Reynolds, John G.; Nunes, Peter; Shields, Sharon J.

    2010-11-09

    A simple and effective system for the colorimetric determination of organic peroxides and hydrogen peroxide. A peroxide pen utilizing a swipe material attached to a polyethylene tube contains two crushable vials. The two crushable vials contain a colorimetric reagent separated into dry ingredients and liquid ingredients. After swiping a suspected substance or surface the vials are broken, the reagent is mixed thoroughly and the reagent is allowed to wick into the swipe material. The presence of organic peroxides or hydrogen peroxide is confirmed by a deep blue color.

  18. Relationship between colorimetric (instrumental) evaluation and consumer-defined beef colour acceptability.

    PubMed

    Holman, Benjamin W B; Mao, Yanwei; Coombs, Cassius E O; van de Ven, Remy J; Hopkins, David L

    2016-11-01

    The relationship between instrumental colorimetric values (L*, a*, b*, the ratio of reflectance at 630nm and 580nm) and consumer perception of acceptable beef colour was evaluated using a web-based survey and standardised photographs of beef m. longissimus lumborum with known colorimetrics. Only L* and b* were found to relate to average consumer opinions of beef colour acceptability. Respondent nationality was also identified as a source of variation in beef colour acceptability score. Although this is a preliminary study with the findings necessitating additional investigation, these results suggest L* and b* as candidates for developing instrumental thresholds for consumer beef colour expectations. PMID:27294520

  19. Rapid Method for Measuring Extracellular Water in Yeast Preparations

    PubMed Central

    Watson, R. W.; Levinson, M. L.

    1967-01-01

    A rapid procedure for the quantitative determination of extracellular water in bulk bakers' yeast was developed on the basis of the solute dilution principle. A reagent is prepared by synthesizing the diazonium ion of p-aminobenzoic acid and coupling it to peptone. This “azopeptone reagent” permits direct colorimetric measurement, which accounts for the rapidity and simplicity of the test. Potential errors due to osmotic effects are avoided by supplementing the reagent with saline and, more importantly, minimizing the duration of contact between reagent and cells. The new method has acceptable accuracy and precision, and may also be suitable for use with other microorganisms. PMID:6043615

  20. Multifunctional Oval Shape Gold Nanoparticle Based Selective Detection of Breast Cancer Cells Using Simple Colorimetric and Highly Sensitive Two-Photon Scattering Assay

    PubMed Central

    Lu, Wentong; Arumugam, Sri Ranjini; Senapati, Dulal; Singh, Anant K.; Arbneshi, Tahir; Yu, Sadia Afrin Khan Hongtao; Ray, Paresh Chandra

    2010-01-01

    Breast cancer is the most common cancer among women and it is the second leading cause of cancer deaths in women today. The key to the effective and ultimately successful treatment of diseases such as cancer is an early and accurate diagnosis. Driven by the need, in this article, we report for the first time a simple colorimetric and highly sensitive two-photon scattering assay for highly selective and sensitive detection of breast cancer SK-BR-3 cell lines in 100-cells/ml level using multifunctional (monoclonal anti-HER2/c-erb-2 antibody and S6 RNA aptamers conjugated) oval shape gold nanoparticle based nanoconjugate. When multifunctional oval shape gold nanoparticles were mixed with breast cancer SK-BR-3 cell line, a distinct color change occurs and two-photon scattering intensity increases by about 13 times. Experimental data with HaCaT non-cancerous cell line, as well as with MDA-MB-231 breast cancer cell line clearly demonstrated that our assay was highly sensitive to SK-BR-3 and it was able to distinguish from other breast cancer cell line which expresses low levels of HER-2. The mechanism of selectivity and assay’s response change, have been discussed. Our experimental results reported here open up a new possibility of rapid, easy and reliable diagnosis of cancer cell lines by monitoring the colorimetric change and measuring TPS intensity from multifunctional gold nanosystems. PMID:20155973

  1. Fluorescence turn-on and colorimetric dual readout assay of glutathione over cysteine based on the fluorescence inner-filter effect of oxidized TMB on TMPyP.

    PubMed

    Jiang, Xiangyu; Geng, Fenghua; Wang, Yongxiang; Liu, Jinhua; Qu, Peng; Xu, Maotian

    2016-07-15

    Quantitative fluorescence turn-on and colorimetric detection of glutathione (GSH) with rapid speed, low cost have attained much attention. Herein, we developed a sensitive fluorescence turn-on and colorimetric sensor for GSH based on the inner-filter effect (IFE), which is the first time to select oxTMB and TMPyP as the IFE absorber and fluorophore pair, respectively. The absorption band of oxTMB matches well with the emission band of TMPyP in the IFE-based fluorescent assay. In the absence of GSH, the absorption peak of oxTMB at 652nm significantly overlaps with the emission of TMPyP, resulting in the efficient IFE and inhibition of the fluorescence of TMPyP. In the presence of GSH, the absorption intensity at 652nm decreases, generating the recovery of the fluorescence of TMPyP. Therefore, this approach is demonstrated to be a novel candidate for detection of GSH, with high sensitivity and selectivity. The linear dynamic range for the concentrations of GSH is between 0.1μM to 20μM along with a limit of detection (LOD) of about 30nM (calculated LOD as 3σ/slope). Finally, this novel sensor was successfully applied for GSH detection in fetal calf serum, and satisfactory recovery was achieved. PMID:26971272

  2. Label-free colorimetric sensing of copper(II) ions based on accelerating decomposition of H2O2 using gold nanorods as an indicator.

    PubMed

    Wang, Shasha; Chen, Zhaopeng; Chen, Ling; Liu, Ruili; Chen, Lingxin

    2013-04-01

    A novel label-free colorimetric strategy was reported for sensitive detection of copper ions (Cu(2+)) by using the decelerating etching of gold nanorods (GNRs). H2O2 was employed as the oxidant for corrosion of GNRs, leading to the decrease of the aspect ratio of GNRs. In the absence of Cu(2+), the redox corrosion of GNRs by H2O2 occurred rapidly, causing the distinct color change of GNRs from bluish green to purplish red. By virtue of the strong and specific catalysis by Cu(2+) of the decomposition of H2O2, the rate of redox corrosion can be decelerated. Relevant experimental parameters, including pH value, concentrations of NaSCN and H2O2, incubation temperature and time were evaluated. Under optimal conditions, our method gave a good linear range of 10-300 nM (R = 0.9985) for Cu(2+) and the detection limit with the naked eye is as low as 10 nM. Thus, the proposed colorimetric sensor is simple, sensitive (4.96 nM) and selective, and it has been successfully applied to detect Cu(2+) in shellfish samples. Moreover, the potential mechanism was also discussed. PMID:23420019

  3. A cost-effective colorimetric assay for phenolic O-methyltransferases and characterization of caffeate 3-O-methyltranferases from Populus trichocarpa

    SciTech Connect

    Bhuiya, M.W.; Liu, C.

    2009-01-01

    S-Adenosyl-L-methionine (AdoMet)-dependent O-methyltransferases (OMTs) catalyze the transmethylation of a variety of phenolics in bacteria, plants, and humans. To rapidly characterize phenolic OMT activities, we adapted Gibbs reagent, the dye originally used for detecting phenols, to develop a convenient assay method for measuring the catalytic properties of enzymatic transmethylation of phenolics. We demonstrated that Gibbs reagent reacted with phenolics yielding distinct absorptive characters that we used to further develop the assay to monitor the reactivities of phenolic OMTs. To validate the method, we identified two caffeate/5-hydroxyferulate 3/5-O-methyltransferases (COMTs) from the black cottonwood, Populus trichocarpa. Together with a few other plant type I OMTs, we demonstrated that our Gibbs reagent-mediated colorimetric assay could reliably determine the functions and kinetic parameters of phenolic OMTs. Because Gibbs reagent reacting with different regioselectively modified phenolics displays different colorimetric properties, the assay method can be used to monitor both substrate specificity and the regioselectivity of phenolic OMTs.

  4. Systematic Assessment of RNA-Seq Quantification Tools Using Simulated Sequence Data

    PubMed Central

    Chandramohan, Raghu; Wu, Po-Yen; Phan, John H.; Wang, May D.

    2016-01-01

    RNA-sequencing (RNA-seq) technology has emerged as the preferred method for quantification of gene and isoform expression. Numerous RNA-seq quantification tools have been proposed and developed, bringing us closer to developing expression-based diagnostic tests based on this technology. However, because of the rapidly evolving technologies and algorithms, it is essential to establish a systematic method for evaluating the quality of RNA-seq quantification. We investigate how different RNA-seq experimental designs (i.e., variations in sequencing depth and read length) affect various quantification algorithms (i.e., HTSeq, Cufflinks, and MISO). Using simulated data, we evaluate the quantification tools based on four metrics, namely: (1) total number of usable fragments for quantification, (2) detection of genes and isoforms, (3) correlation, and (4) accuracy of expression quantification with respect to the ground truth. Results show that Cufflinks is able to use the largest number of fragments for quantification, leading to better detection of genes and isoforms. However, HTSeq produces more accurate expression estimates. Moreover, each quantification algorithm is affected differently by varying sequencing depth and read length, suggesting that the selection of quantification algorithms should be application-dependent.

  5. Validated Colorimetric Assay of Clonidine Hydrochloride from Pharmaceutical Preparations.

    PubMed

    Corciova, Andreia

    2016-01-01

    Clonidine hydrochloride is an antihypertensive agent used for migraine prophylaxis, attention deficit hyperactivity disorder, menopausal flushing and Tourette syndrome. The quantity of the active substance in pharmaceutical preparations must be within specific limits, in agreement with the respective label claim. Therefore, the aim of this study was to establish the conditions for two spectrophotometric methods for clonidine determination, based on the formation of the ion pair complex between clonidine hydrochloride and thymol blue/bromophenol blue. A Jasco UV-Vis 530 spectrophotometer was used for the analysis and the maxim absorbance was measured at 418 nm/448 nm against blank solution. After validation, the methods were used for quantification of clonidine hydrochloride in two commercial samples (tablets). The recovery of active substance varies between 98.06 and 100.13 % without interferences from the excipients. PMID:27610155

  6. Validated Colorimetric Assay of Clonidine Hydrochloride from Pharmaceutical Preparations

    PubMed Central

    Corciova, Andreia

    2016-01-01

    Clonidine hydrochloride is an antihypertensive agent used for migraine prophylaxis, attention deficit hyperactivity disorder, menopausal flushing and Tourette syndrome. The quantity of the active substance in pharmaceutical preparations must be within specific limits, in agreement with the respective label claim. Therefore, the aim of this study was to establish the conditions for two spectrophotometric methods for clonidine determination, based on the formation of the ion pair complex between clonidine hydrochloride and thymol blue/bromophenol blue. A Jasco UV-Vis 530 spectrophotometer was used for the analysis and the maxim absorbance was measured at 418 nm/448 nm against blank solution. After validation, the methods were used for quantification of clonidine hydrochloride in two commercial samples (tablets). The recovery of active substance varies between 98.06 and 100.13 % without interferences from the excipients. PMID:27610155

  7. Design of a dual-function peptide probe as a binder of angiotensin II and an inducer of silver nanoparticle aggregation for use in label-free colorimetric assays.

    PubMed

    Okochi, Mina; Kuboyama, Masashi; Tanaka, Masayoshi; Honda, Hiroyuki

    2015-09-01

    Label-free colorimetric assays using metallic nanoparticles have received much recent attention, for their application in simple and sensitive methods for detection of biomolecules. Short peptide probes that can bind to analyte biomolecules are attractive ligands in molecular nanotechnology; however, identification of biological recognition motifs is usually based on trial-and-error experiments. Herein, a peptide probe was screened for colorimetric detection of angiotensin II (Ang II) using a mechanism for non-crosslinking aggregation of silver nanoparticles (AgNPs). The dual-function peptides, which bind to the analyte and induce AgNP aggregation, were identified using a two-step strategy: (1) screening of an Ang II-binding peptide from an Ang II receptor sequence library, using SPOT technology, which enable peptides synthesis on cellulose membranes via an Fmoc method and (2) selection of peptide probes that effectively induce aggregation of AgNPs using a photolinker modified peptide array. Using the identified peptide probe, KGKNKRRR, aggregation of AgNPs was detected by observation of a pink color in the absence of Ang II, whereas AgNPs remained dispersed in the presence of Ang II (yellow). The color changes were not observed in the presence of other hormone molecules. Ang II could be detected within 15 min, with a detection limit of 10 µM, by measuring the ratio of absorbance at 400 nm and 568 nm; the signal could also be observed with the naked eye. These data suggest that the peptide identified here could be used as a probe for simple and rapid colorimetric detection of Ang II. This strategy for the identification of functional peptides shows promise for the development of colorimetric detection of various diagnostically important biomolecules. PMID:26003717

  8. Colorimetric Detection of Escherichia coli Based on the Enzyme-Induced Metallization of Gold Nanorods.

    PubMed

    Chen, Juhong; Jackson, Angelyca A; Rotello, Vincent M; Nugen, Sam R

    2016-05-01

    A novel enzyme-induced metallization colorimetric assay is developed to monitor and measure beta-galactosidase (β-gal) activity, and is further employed for colorimetric bacteriophage (phage)-enabled detection of Escherichia coli (E. coli). This assay relies on enzymatic reaction-induced silver deposition on the surface of gold nanorods (AuNRs). In the presence of β-gal, the substrate p-aminophenyl β-d-galactopyranoside is hydrolyzed to produce p-aminophenol (PAP). Reduction of silver ions by PAP generates a silver shell on the surface of AuNRs, resulting in the blue shift of the longitudinal localized surface plasmon resonance peak and multicolor changes of the detection solution from light green to orange-red. Under optimized conditions, the detection limit for β-gal is 128 pM, which is lower than the conventional colorimetric assay. Additionally, the assay has a broader dynamic range for β-gal detection. The specificity of this assay for the detection of β-gal is demonstrated against several protein competitors. Additionally, this technique is successfully applied to detect E. coli bacteria cells in combination with bacteriophage infection. Due to the simplicity and short incubation time of this enzyme-induced metallization colorimetric method, the assay is well suited for the detection of bacteria in low-resource settings. PMID:26997252

  9. Potassium sorbate residue levels and persistence in citrus fruit as detected by a simple colorimetric method

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Postharvest applications of potassium sorbate (PS) to fresh citrus fruit control fungal decay pathogens, such as Penicillium digitatum, cause of green mold. Although PS effectiveness has been examined repeatedly, little is known about PS residues. A colorimetric method that employed extraction of th...

  10. A nanoplasmonic probe as a triple channel colorimetric sensor array for protein discrimination.

    PubMed

    Mao, Jinpeng; Lu, Yuexiang; Chang, Ning; Yang, Jiaoe; Yang, Jiacheng; Zhang, Sichun; Liu, Yueying

    2016-06-20

    The salt-induced aggregation, nanoparticle regrowth and self-assembly behaviors of gold nanoparticles (AuNPs) and DNA conjugates could be changed after interaction with different proteins, generating various color changes and a unique fingerprint pattern for each protein. The triple-channel colorimetric signals have been employed for protein discrimination with the naked eye. PMID:27228956

  11. A novel colorimetric sensor for Hg(2+) based on hybridization chain reaction and silver nanowire amplification.

    PubMed

    Tang, Shurong; Tong, Ping; Wang, Meili; Chen, Jinghua; Li, Guangwen; Zhang, Lan

    2015-10-18

    Through the silver ion catalysis to form colored KMnO4, and combined with the DNA hybridization chain reaction and silver nanowire for signal amplification, a highly sensitive and selective colorimetric sensor has been developed for the detection of Hg(2+). PMID:26313655

  12. Pyridoxamine-5-phosphate enzyme-linked immune mass spectrometric assay substrate for linear absolute quantification of alkaline phosphatase to the yoctomole range applied to prostate specific antigen.

    PubMed

    Florentinus-Mefailoski, Angelique; Marshall, John G

    2014-11-01

    There is a need to measure proteins that are present in concentrations below the detection limits of existing colorimetric approaches with enzyme-linked immunoabsorbent assays (ELISA). The powerful enzyme alkaline phosphatase conjugated to the highly specific bacterial protein streptavidin binds to biotinylated macromolecules like proteins, antibodies, or other ligands and receptors with a high affinity. The binding of the biotinylated detection antibody, with resulting amplification of the signal by the catalytic production of reporter molecules, is key to the sensitivity of ELISA. The specificity and amplification of the signal by the enzyme alkaline phosphatase in ELISA together with the sensitivity of liquid chromatography electrospray ionization and mass spectrometry (LC-ESI-MS) to detect femtomole to picomole amounts of reporter molecules results in an ultrasensitive enzyme-linked immune mass spectrometric assay (ELIMSA). The novel ELIMSA substrate pyridoxamine-5-phosphate (PA5P) is cleaved by the enzyme alkaline phosphatase to yield the basic and hydrophilic product pyridoxamine (PA) that elutes rapidly with symmetrical peaks and a flat baseline. Pyridoxamine (PA) and (13)C PA were both observed to show a linear relationship between log ion intensity and quantity from picomole to femtomole amounts by liquid chromatography-electrospray ionization and mass spectrometry. Four independent methods, (i) internal (13)C isotope PA dilution curves, (ii) internal (13)C isotope one-point calibration, (iii) external PA standard curve, and (iv) external (13)C PA standard curve, all agreed within 1 digit in the same order of magnitude on the linear quantification of PA. Hence, a mass spectrometer can be used to robustly detect 526 ymol of the alkaline phosphatase streptavidin probe and accurately quantify zeptomole amounts of PSA against log linear absolute standard by micro electrospray on a simple ion trap. PMID:25259405

  13. Multispectral image analysis for algal biomass quantification.

    PubMed

    Murphy, Thomas E; Macon, Keith; Berberoglu, Halil

    2013-01-01

    This article reports a novel multispectral image processing technique for rapid, noninvasive quantification of biomass concentration in attached and suspended algae cultures. Monitoring the biomass concentration is critical for efficient production of biofuel feedstocks, food supplements, and bioactive chemicals. Particularly, noninvasive and rapid detection techniques can significantly aid in providing delay-free process control feedback in large-scale cultivation platforms. In this technique, three-band spectral images of Anabaena variabilis cultures were acquired and separated into their red, green, and blue components. A correlation between the magnitude of the green component and the areal biomass concentration was generated. The correlation predicted the biomass concentrations of independently prepared attached and suspended cultures with errors of 7 and 15%, respectively, and the effect of varying lighting conditions and background color were investigated. This method can provide necessary feedback for dilution and harvesting strategies to maximize photosynthetic conversion efficiency in large-scale operation. PMID:23554374

  14. Rapid detection and quantification of impact damage in composite structures

    NASA Technical Reports Server (NTRS)

    Zalameda, Joseph N.; Farley, Gary; Smith, Barry T.

    1991-01-01

    NDE results from thermographic and volumetric ultrasonic techniques are presented to illustrate the multidisciplinary NDE approach to impact-damage detection in such composite structures as are increasingly prevalent in helicopters. Attention is given to both flat-panel and 'y-stiffened' panel specimens; these were fabricated either with kevlar or carbon fiber through-the-thickness reinforcements. While thermal inspection identifies impact damage, volumetric imaging quantifies the impact-generated delaminations through the volume of the structure.

  15. RAPID CONCENTRATION, DETECTION, AND QUANTIFICATION OF PATHOGENS IN DRINKING WATER

    EPA Science Inventory

    1. Concentration of microbes in water using a lanthanum-based colloidal destabilization method and determine bacteria concentrations by fluorescence-based oxygen microrespirometry
     
    Lanthanum-based salts (e.g., ...

  16. Nanoparticle-catalyzed reductive bleaching for fabricating turn-off and enzyme-free amplified colorimetric bioassays.

    PubMed

    Li, Wei; Qiang, Weibing; Li, Jie; Li, Hui; Dong, Yifan; Zhao, Yaju; Xu, Danke

    2014-01-15

    Nanoparticle-catalyzed reductive bleaching reactions of colored substrates are emerging as a class of novel indicator reactions for fabricating enzyme-free amplified colorimetric biosensing (turn-off mode), which are exactly opposite to the commonly used oxidative coloring processes of colorless substrates in traditional enzyme-catalyzed amplified colorimetric bioassays (turn-on mode). In this work, a simple theoretical analysis shows that the sensitivity of this colorimetric bioassay can be improved by increasing the amplification factor (kcatΔt), or enhancing the binding affinity between analyte and receptor (Kd), or selecting the colored substrates with high extinction coefficients (ε). Based on this novel strategy, we have developed a turn-off and cost-effective amplified colorimetric thrombin aptasensor. This aptasensor made full use of sandwich binding of two affinity aptamers for increased specificity, magnetic particles for easy separation and enrichment, and gold nanoparticle (AuNP)-catalyzed reductive bleaching reaction to generate the amplified colorimetric signal. With 4-nitrophenol (4-NP) as the non-dye colored substrate, colorimetric bioassay of thrombin was achieved by the endpoint method with a detection limit of 91pM. In particular, when using methylene blue (MB) as the substrate, for the first time, a more convenient and efficient kinetic-based colorimetric thrombin bioassay was achieved without the steps of acidification termination and magnetic removal of particles, with a low detection limit of 10pM, which was superior to the majority of the existing colorimetric thrombin aptasensors. The proposed colorimetric protocol is expected to hold great promise in field analysis and point-of-care applications. PMID:23962710

  17. Intrinsic enzyme mimicking activity of gold nanoclusters upon visible light triggering and its application for colorimetric trypsin detection.

    PubMed

    Wang, Guang-Li; Jin, Lu-Yi; Dong, Yu-Ming; Wu, Xiu-Ming; Li, Zai-Jun

    2015-02-15

    In this research, a novel enzyme mimetics based on the photochemical property of gold nanoclusters was demonstrated. It was found that the bovine serum albumin (BSA) stabilized red or blue emitting gold nanoclusters (Au NCs) exhibited enzyme-like activity under visible light irradiation. The BSA-Au NCs had better stability against stringent conditions compared to natural enzyme. In addition, the photostimulated enzyme mimetics of BSA-Au NCs showed several unprecedented advantages over natural peroxidase or other existing alternatives based on nanomaterials, such as the independence of hydrogen peroxide on activity and the easily regulated activity by light irradiation. The mechanism of the photoresponsive enzyme-like activity of BSA-Au NCs was investigated. The photoactivated BSA-Au NCs was designed to develop a facile, cheap, and rapid colorimetric assay to detect trypsin through trypsin digestion of the protein template of BSA-stabilized Au NCs. The limit of detection for trypsin was 0.6 μg/mL, which was much lower than the average level of trypsin in patient's urine or serum. PMID:25310483

  18. Colorimetric detection of Bi (III) in water and drug samples using pyridine-2,6-dicarboxylic acid modified silver nanoparticles

    NASA Astrophysics Data System (ADS)

    Mohammadi, Somayeh; Khayatian, Gholamreza

    2015-09-01

    A new selective, simple, fast and sensitive method is developed for sensing assay of Bi (III) using pyridine-2,6-dicarboxylic acid or dipicolinic acid (DPA) modified silver nanoparticles (DPA-AgNPs). Silver nanoparticles (AgNPs) were synthesized by reducing silver nitrate (AgNO3) with sodium borohydride (NaBH4) in the presence of DPA. Bismuth detection is based on color change of nanoparticle solution from yellow to red that is induced in the presence of Bi (III). Aggregation of DPA-AgNPs has been confirmed with UV-vis absorption spectra and transmission electron microscopy (TEM) images. Under the optimized conditions, a good linear relationship (correlation coefficient r = 0.995) is obtained between the absorbance ratio (A525/A390) and the concentration of Bi (III) in the 0.40-8.00 μM range. This colorimetric probe allows Bi (III) to be rapidly quantified with a 0.01 μM limit of detection. The present method successfully applied to determine bismuth in real water and drug samples. Recoveries of water samples were in the range of 91.2-99.6%.

  19. Microfluidic toner-based analytical devices: disposable, lightweight, and portable platforms for point-of-care diagnostics with colorimetric detection.

    PubMed

    Oliveira, Karoliny Almeida; de Souza, Fabrício Ribeiro; de Oliveira, Cristina Rodrigues; da Silveira, Lucimeire Antonelli; Coltro, Wendell Karlos Tomazelli

    2015-01-01

    This chapter describes the development of microfluidic toner-based analytical devices (μTADs) to perform clinical diagnostics using a scanner or cell-phone camera. μTADs have been produced in a platform composed of polyester and toner by the direct-printing technology (DPT) in a matter of minutes. This technology offers simplicity and versatility, and it does not require any sophisticated instrumentation. Toner-based devices integrate the current generation of disposable analytical devices along paper-based chips. The cost of one μTAD has been estimated to be lower than $0.10. In addition, these platforms are lightweight and portable thus enabling their use for point-of-care applications. In the last 5 years, great efforts have been dedicated to spread out the use of μTADs in bioassays. The current chapter reports the fabrication of printed microplates and integrated microfluidic toner-based devices for dengue diagnostics and rapid colorimetric assays with clinically relevant analytes including cholesterol, triglycerides, total proteins, and glucose. The use of μTADs associated with cell-phone camera may contribute to the health care, in special, to people housed in developing regions or with limited access to clinics and hospitals. PMID:25626533

  20. A universal probe design for colorimetric detection of single-nucleotide variation with visible readout and high specificity

    PubMed Central

    Chen, Xueping; Zhou, Dandan; Shen, Huawei; Chen, Hui; Feng, Wenli; Xie, Guoming

    2016-01-01

    Single-nucleotide variation (SNV) is a crucial biomarker for drug resistance-related detection in cancer and bacterial infection. However, the unintended binding of DNA probes limits the specificity of SNV detection, and the need for redesigned sequences compromise the universality of SNV assay. Herein, we demonstrated a universal and low-cost assay for the colorimetric discrimination of drug-resistance related point mutation. By the use of a universal DNA probe and a split G-quadruplex, the signal could be recognized by naked eye at room temperature. The DNA probe was used as a signal reporter which not only improved the universality, but also enabled high specificity of probe hybridization. This assay was successfully applied in the detection of cancer-related SNV in the epidermal growth factor receptor (EGFR) gene, kirsten rat sarcoma viral oncogene homologue (KRAS), and tuberculosis drug-resistance related point mutation in RNA polymerase beta subunit gene (rpoB) with high specificity and visible readout. This method was simple, rapid, high-throughput and effective, which was suitable for point-of-care applications. PMID:26830326

  1. Detection of measles, mumps and rubella viruses by immuno-colorimetric assay and its application in focus reduction neutralization tests.

    PubMed

    Vaidya, Sunil R; Kumbhar, Neelakshi S; Bhide, Vandana S

    2014-12-01

    Measles, mumps and rubella are vaccine-preventable diseases; however limited epidemiological data are available from low-income or developing countries. Thus, it is important to investigate the transmission of these viruses in different geographical regions. In this context, a cell culture-based rapid and reliable immuno-colorimetric assay (ICA) was established and its utility studied. Twenty-three measles, six mumps and six rubella virus isolates and three vaccine strains were studied. Detection by ICA was compared with plaque and RT-PCR assays. In addition, ICA was used to detect viruses in throat swabs (n = 24) collected from patients with suspected measles or mumps. Similarly, ICA was used in a focus reduction neutralization test (FRNT) and the results compared with those obtained by a commercial IgG enzyme immuno assay. Measles and mumps virus were detected 2 days post-infection in Vero or Vero-human signaling lymphocytic activation molecule cells, whereas rubella virus was detected 3 days post-infection in Vero cells. The blue stained viral foci were visible by the naked eye or through a magnifying glass. In conclusion, ICA was successfully used on 35 virus isolates, three vaccine strains and clinical specimens collected from suspected cases of measles and mumps. Furthermore, an application of ICA in a neutralization test (i.e., FRNT) was documented; this may be useful for sero-epidemiological, cross-neutralization and pre/post-vaccine studies. PMID:25244651

  2. Preparation of CuO/ZnO nanocomposite and its application as a cysteine/homocysteine colorimetric and fluorescence detector.

    PubMed

    Šimšíková, Michaela; Čechal, Jan; Zorkovská, Anna; Antalík, Marián; Šikola, Tomáš

    2014-11-01

    Cysteine and homocysteine play a crucial role in many biological functions but abnormal levels of these amino acids may lead to various forms of pathogenesis. Therefore, selective and easy-to-use methods for the detection of cysteine and homocysteine are essential for the early diagnosis of developing diseases. In this paper we report on a rapid, straightforward and highly selective method for the detection of cysteine (Cys) and homocysteine (Hcy) which uses a CuO/ZnO nanocomposite as a dual colorimetric and fluorometric assay. The presence of Cys and Hcy in a solution of these nanorods (NRs) induces a change in its color from light blue to dark grey which is visible to the naked eye. This is accompanied by a blue shift in the absorption spectra from 725 nm to 650 nm and a decrease in the intensity of CuO/ZnO nanocomposite emission. These changes are ascribed to the reduction of Cu(II) to Cu(0), and the oxidation of cysteine (homocysteine) and subsequent formation of the disulfide bond. This novel assay method does not respond to any other amino-acid which is present in living organisms; therefore the selective determination of cysteine (homocysteine) with a lower analyte limit of 40 μM (4.8 μg mL(-1)) can be carried out in aqueous solutions without the need for any sophisticated instrumentation, fluorophore molecules or complicated procedures. PMID:25465753

  3. Dextrin-mediated synthesis of Ag NPs for colorimetric assays of Cu(2+) ion and Au NPs for catalytic activity.

    PubMed

    Bankura, Kalipada; Rana, Dipak; Mollick, Md Masud Rahaman; Pattanayak, Sutanuka; Bhowmick, Biplab; Saha, Nayan Ranjan; Roy, Indranil; Midya, Tarapada; Barman, Gadadhar; Chattopadhyay, Dipankar

    2015-09-01

    A facile one-pot approach for rapid synthesis of silver and gold nanoparticles (Ag NPs and Au NPs) with narrow size distribution and good stability was described by reducing silver nitrate and chloroauric acid with polysaccharide dextrin. Here, dextrin was used as both a reducing and stabilizing agent for synthesis of NPs. The as-synthesized Ag NPs and Au NPs were characterized by UV-visible absorption spectroscopy, transmission electron microscopy (TEM) and X-ray diffraction (XRD). The Ag NPs and Au NPs exhibited an absorption maxima at 404 and 547 nm respectively. TEM images showed NPs in the range of 8-28 nm. The crystallinity of the NPs was measured by XRD analysis. Furthermore, the as-prepared Ag NPs revealed colorimetric sensor property for detection of Cu(2+) ions based on changes in absorbance resulting from metal ion-induced aggregation of NPs or direct deposition of metal ions onto NPs. The as-prepared Au NPs exhibited a notable catalytic activity toward the reduction of 4-nitrophenol to 4-aminophenol in the presence of NaBH4. PMID:26143120

  4. Colorimetric detection of Bi (III) in water and drug samples using pyridine-2,6-dicarboxylic acid modified silver nanoparticles.

    PubMed

    Mohammadi, Somayeh; Khayatian, Gholamreza

    2015-09-01

    A new selective, simple, fast and sensitive method is developed for sensing assay of Bi (III) using pyridine-2,6-dicarboxylic acid or dipicolinic acid (DPA) modified silver nanoparticles (DPA-AgNPs). Silver nanoparticles (AgNPs) were synthesized by reducing silver nitrate (AgNO3) with sodium borohydride (NaBH4) in the presence of DPA. Bismuth detection is based on color change of nanoparticle solution from yellow to red that is induced in the presence of Bi (III). Aggregation of DPA-AgNPs has been confirmed with UV-vis absorption spectra and transmission electron microscopy (TEM) images. Under the optimized conditions, a good linear relationship (correlation coefficient r=0.995) is obtained between the absorbance ratio (A525/A390) and the concentration of Bi (III) in the 0.40-8.00 μM range. This colorimetric probe allows Bi (III) to be rapidly quantified with a 0.01 μM limit of detection. The present method successfully applied to determine bismuth in real water and drug samples. Recoveries of water samples were in the range of 91.2-99.6%. PMID:25919329

  5. In situ formation of phosphorescent molecular gold(I) cluster in a macroporous polymer film to achieve colorimetric cyanide sensing.

    PubMed

    Zong, Chenghua; Zheng, Li Rong; He, Wenhui; Ren, Xiaoyan; Jiang, Chunhuan; Lu, Lehui

    2014-02-01

    A highly phosphorescent molecular Au(I) cluster capable of rapid, sensitive, and selective detection of cyanide has been successfully fabricated. The origin of the outstanding sensing performance of the molecular Au(I) cluster toward cyanide is justified by X-ray absorption near edge structure (XANES) and extended X-ray absorption fine structure (EXAFS) analyses. The response mechanism employed with the molecular Au(I) cluster and the cost-effectiveness in cyanide detection affords several key sensor features, making this molecular Au(I) cluster-based sensor unique compared to other cyanide sensing schemes. Importantly, by exploring the phosphorescent properties of the molecular Au(I) cluster in solid state, we demonstrate the first example of the molecular gold(I) cluster-based macroporous sensing film for colorimetric detection of cyanide in complex samples, including red wine, coffee, juice, and soil. Remarkably, the as-prepared sensing film inherits the sensing ability of the molecular Au(I) cluster, and offers a high mechanical flexibility and novel opportunities for real-time monitoring cyanide release in cassava manufacturing. PMID:24397707

  6. Quantification of Hepcidin-related Iron Accumulation in the Rat Liver.

    PubMed

    Böser, Preethne; Mordashova, Yulia; Maasland, Mark; Trommer, Isabel; Lorenz, Helga; Hafner, Mathias; Seemann, Dietmar; Mueller, Bernhard K; Popp, Andreas

    2016-02-01

    Hepcidin was originally detected as a liver peptide with antimicrobial activity and it functions as a central regulator in the systemic iron metabolism. Consequently suppression of hepcidin leads to iron accumulation in the liver. AbbVie developed a monoclonal antibody ([mAb]; repulsive guidance molecule [RGMa/c] mAb) that downregulates hepcidin expression by influencing the RGMc/bone morphogenetic protein (BMP)/neogenin receptor complex and causes iron deposition in the liver. In a dose range finding study with RGMa/c mAb, rats were treated with different dose levels for a total of 4 weekly doses. The results of this morphometric analysis in the liver showed that iron accumulation is not homogenous between liver lobes and the left lateral lobe was the most responsive lobe in the rat. Quantitative hepcidin messenger RNA analysis showed that the left lateral lobe was the most responsive lobe showing hepcidin downregulation with increasing antibody dose. In addition, the morphometric analysis had higher sensitivity than the chemical iron extraction and quantification using a colorimetric assay. In conclusion, the Prussian blue stain in combination with semi-quantitative and quantitative morphometric analysis is the most reliable method to demonstrate iron accumulation in the liver compared to direct measurement of iron in unfixed tissue using a colorimetric assay. PMID:26839325

  7. Surface-plasmon-based colorimetric detection of Cu(II) ions using label-free gold nanoparticles in aqueous thiosulfate systems

    NASA Astrophysics Data System (ADS)

    Tripathy, Suraj Kumar; Woo, Ju Yeon; Han, Chang-Soo

    2012-08-01

    We report colorimetric, label-free and non-aggregation-based gold nanoparticle (AuNP) probes for the highly selective detection of Cu(II) ions in aqueous environments. This detection scheme relies on the ability of Cu(II) ions to catalyze the leaching of gold at room temperature in the presence of thiosulfate species and ammonia. This simple and cost-effective probe provides rapid detection of Cu(II) ions at concentrations as low as 10 ppm. A similar detection method using AuNPs in ammonia-free thiosulfate solution is also viable at moderate reaction temperature (50 °C). The ammonia-free method also leads to marked damping and red-shifting of the surface plasmon resonance signal of the AuNP dispersion. The two methods clearly differ in the nature of the surface plasmon damping phenomenon, and their working mechanisms are plausibly explained based on the experimental investigations.

  8. Carbon dots-assisted colorimetric and fluorometric dual-mode protocol for acetylcholinesterase activity and inhibitors screening based on the inner filter effect of silver nanoparticles.

    PubMed

    Zhao, Dan; Chen, Chuanxia; Sun, Jian; Yang, Xiurong

    2016-06-01

    In this work, we proposed an original and versatile dual-readout (colorimetric and fluorometric) protocol by means of silver nanoparticles (AgNPs) and fluorescent carbon dots (CDs), which was amenable to rapid, ultrasensitive assay of acetylcholinesterase (AChE) activity and its inhibitors. The sensing mechanism was based on the non-fluorescence state of CDs resulting from the inner filter effect (IFE) of AgNPs and the specific AChE-catalyzed hydrolysis of acetylthiocholine (ATCh) into thiocholine (TCh). Herein, the generated positively-charged and thiol-bearing TCh at trace concentration levels could trigger the aggregation of AgNPs through the well-known electrostatic and Ag-SH interactions, thereby turning the sensing solutions grey and recovering the IFE-quenched fluorescence simultaneously. Furthermore, the existence of IFE mechanism was conceivably confirmed by combining the zeta potentials, fluorescence spectra, UV-vis spectra, fluorescence lifetime and TEM measurements. As far as we know, the present study has reported the first dual-mode proposal for assessing AChE activity by using a CDs-based IFE sensing strategy, where the detection limit was as low as 0.021 mU mL(-1) and 0.016 mU mL(-1) by colorimetric and fluorometric measurements, respectively. On the other hand, the proposed assay was feasible to screen AChE inhibitors such as tacrine and carbaryl. Meanwhile, this rationally designed dual-mode sensing platform featured simplicity, rapidity, flexibility and diversity, which was demonstrated by the quantitative detection of spiked carbaryl in apple juice samples with satisfactory results. PMID:27099097

  9. Absorbance amplification using chromophore-nanoplasmon coupling for ultrasensitive protein quantification.

    PubMed

    Seo, Sujin; Edwards, Lonna; Logan Liu, Gang

    2015-10-01

    A plasmonic nanoscale Lycurgus cup array (nanoLCA), via near-field interaction with chromophores in commercial colorimetric biochemical assays, can drastically enhance assay sensitivity by over 2 orders of magnitude. A 96-microwell plate modified by placing the plasmonic nanoLCA on the well-bottom was used with the commercial Bradford protein quantification assay. Plasmons on the nanoLCA serve as an energy donor to matched resonance chromophores, and the near-field plasmonic energy coupling effect results in an increase in absorbance value at the plasmonic resonance wavelength. Even with a 5.1-fold reduced sample volume, a limit of detection enhancement factor of 200 is accomplished using the nanoLCA compared to using the conventional Bradford assay without plasmon aid. The nanoLCA-microplate sensing platform is readily scalable to 384- or 1536-microwell plates, which further reduces the sensing volume and boosts detection throughput with the enhanced sensitivity. PMID:26284911

  10. 4-Aminothiophenol functionalized gold nanoparticle-based colorimetric sensor for the determination of nitramine energetic materials.

    PubMed

    Üzer, Ayşem; Can, Ziya; Akın, Ilknur; Erçağ, Erol; Apak, Reşat

    2014-01-01

    The heterocyclic nitramine compounds, hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX), are two most important military-purpose high explosives. Differentiation of RDX and HMX with colorimetric methods of determination has not yet been made because of their similar chemical structures. In this study, a sensitive colorimetric method for the determination of RDX and HMX was proposed on the basis of differential kinetics in the hydrolysis of the two compounds (yielding nitrite as a product) followed by their colorimetric determination using 4-aminothiophenol (4-ATP) modified gold nanoparticles (AuNPs) and naphthylethylene diamine (NED) as coupling agent for azo-dye formation, abbreviated as "4-ATP-AuNP+NED" colorimetric method. After alkaline hydrolysis in a 1 M Na2CO3 + 0.04 M NaOH mixture solution at room temperature, only RDX (but not HMX) was hydrolyzed to give a sufficient colorimetric response in neutralized solution, the molar absorptivity (ε) at 565 nm and the limit of detection (LOD) for RDX being (17.6 ± 1.3) × 10(3) L mol(-1) cm(-1) and 0.55 μg mL(-1), respectively. On the other hand, hot water bath (at 60 °C) hydrolysis enabled both nitramines, RDX and HMX, to give substantial colorimetric responses; i.e., ε and LOD for RDX were (32.8 ± 0.5) × 10(3) L mol(-1)cm(-1) and 0.20 μg mL(-1) and for HMX were (37.1 ± 2.8) × 10(3) L mol(-1)cm(-1) and 0.24 μg mL(-1), respectively. Unlike other AuNP-based nitrite sensors in the literature showing absorbance quenching within a relatively narrow concentration range, the developed sensor operated with an absorbance increase over a wide range of nitrite. Synthetic mixtures of (RDX + HMX) gave additive responses, and the proposed method was statistically validated against HPLC using nitramine mixtures. PMID:24299426

  11. Statistical Quantification of Methylation Levels by Next-Generation Sequencing

    PubMed Central

    Wu, Guodong; Yi, Nengjun; Absher, Devin; Zhi, Degui

    2011-01-01

    Background/Aims Recently, next-generation sequencing-based technologies have enabled DNA methylation profiling at high resolution and low cost. Methyl-Seq and Reduced Representation Bisulfite Sequencing (RRBS) are two such technologies that interrogate methylation levels at CpG sites throughout the entire human genome. With rapid reduction of sequencing costs, these technologies will enable epigenotyping of large cohorts for phenotypic association studies. Existing quantification methods for sequencing-based methylation profiling are simplistic and do not deal with the noise due to the random sampling nature of sequencing and various experimental artifacts. Therefore, there is a need to investigate the statistical issues related to the quantification of methylation levels for these emerging technologies, with the goal of developing an accurate quantification method. Methods In this paper, we propose two methods for Methyl-Seq quantification. The first method, the Maximum Likelihood estimate, is both conceptually intuitive and computationally simple. However, this estimate is biased at extreme methylation levels and does not provide variance estimation. The second method, based on Bayesian hierarchical model, allows variance estimation of methylation levels, and provides a flexible framework to adjust technical bias in the sequencing process. Results We compare the previously proposed binary method, the Maximum Likelihood (ML) method, and the Bayesian method. In both simulation and real data analysis of Methyl-Seq data, the Bayesian method offers the most accurate quantification. The ML method is slightly less accurate than the Bayesian method. But both our proposed methods outperform the original binary method in Methyl-Seq. In addition, we applied these quantification methods to simulation data and show that, with sequencing depth above 40–300 (which varies with different tissue samples) per cleavage site, Methyl-Seq offers a comparable quantification

  12. Advancing agricultural greenhouse gas quantification*

    NASA Astrophysics Data System (ADS)

    Olander, Lydia; Wollenberg, Eva; Tubiello, Francesco; Herold, Martin

    2013-03-01

    1. Introduction Better information on greenhouse gas (GHG) emissions and mitigation potential in the agricultural sector is necessary to manage these emissions and identify responses that are consistent with the food security and economic development priorities of countries. Critical activity data (what crops or livestock are managed in what way) are poor or lacking for many agricultural systems, especially in developing countries. In addition, the currently available methods for quantifying emissions and mitigation are often too expensive or complex or not sufficiently user friendly for widespread use. The purpose of this focus issue is to capture the state of the art in quantifying greenhouse gases from agricultural systems, with the goal of better understanding our current capabilities and near-term potential for improvement, with particular attention to quantification issues relevant to smallholders in developing countries. This work is timely in light of international discussions and negotiations around how agriculture should be included in efforts to reduce and adapt to climate change impacts, and considering that significant climate financing to developing countries in post-2012 agreements may be linked to their increased ability to identify and report GHG emissions (Murphy et al 2010, CCAFS 2011, FAO 2011). 2. Agriculture and climate change mitigation The main agricultural GHGs—methane and nitrous oxide—account for 10%-12% of anthropogenic emissions globally (Smith et al 2008), or around 50% and 60% of total anthropogenic methane and nitrous oxide emissions, respectively, in 2005. Net carbon dioxide fluxes between agricultural land and the atmosphere linked to food production are relatively small, although significant carbon emissions are associated with degradation of organic soils for plantations in tropical regions (Smith et al 2007, FAO 2012). Population growth and shifts in dietary patterns toward more meat and dairy consumption will lead to

  13. Iodide-Responsive Cu-Au Nanoparticle-Based Colorimetric Platform for Ultrasensitive Detection of Target Cancer Cells.

    PubMed

    Ye, Xiaosheng; Shi, Hui; He, Xiaoxiao; Wang, Kemin; He, Dinggeng; Yan, Lv'an; Xu, Fengzhou; Lei, Yanli; Tang, Jinlu; Yu, Yanru

    2015-07-21

    Colorimetric analysis is promising in developing facile, fast, and point-of-care cancer diagnosis techniques, but the existing colorimetric cancer cell assays remain problematic because of dissatisfactory sensitivity as well as complex probe design or synthesis. To solve the problem, we here present a novel colorimetric analytical strategy based on iodide-responsive Cu-Au nanoparticles (Cu-Au NPs) combined with the iodide-catalyzed H2O2-TMB (3,3,5,5-tetramethylbenzidine) reaction system. In this strategy, bimetallic Cu-Au NPs prepared with an irregular shape and a diameter of ∼15 nm could chemically absorb iodide, thus indirectly inducing colorimetric signal variation of the H2O2-TMB system. By further utilizing its property of easy biomolecule modification, a versatile colorimetric platform was constructed for detection of any target that could cause the change of Cu-Au NPs concentration via molecular recognition. As proof of concept, an analysis of human leukemia CCRF-CEM cells was performed using aptamer Sgc8c-modified Cu-Au NPs as the colorimetric probe. Results showed that Sgc8c-modified Cu-Au NPs successfully achieved a simple, label-free, cost-effective, visualized, selective, and ultrasensitive detection of cancer cells with a linear range from 50 to 500 cells/mL and a detection limit of 5 cells in 100 μL of binding buffer. Moreover, feasibility was demonstrated for cancer cell analysis in diluted serum samples. The iodide-responsive Cu-Au NP-based colorimetric strategy might not only afford a new design pattern for developing cancer cell assays but also greatly extend the application of the iodide-catalyzed colorimetric system. PMID:26100583

  14. Label-free colorimetric detection of biological thiols based on target-triggered inhibition of photoinduced formation of AuNPs

    NASA Astrophysics Data System (ADS)

    Lim Jung, Ye; Park, Jung Hun; Kim, Moon Il; Park, Hyun Gyu

    2016-02-01

    A label-free colorimetric method for the detection of biological thiols (biothiols) was developed. This method is based on prevention of the photoinduced reduction of auric ions (Au(III)) in the presence of amino acids (acting as a reducing agent) by biothiols; the photoinduced reduction is inhibited due to the strong interaction of the biothiols with Au(III). In this method, the sample was first incubated in an assay solution containing Au(III) and threonine; the sample solution was then exposed to 254 nm UV light. For samples without biothiols, this process led to the photoreduction of Au(III) followed by growth of gold nanoparticles accompanied by the visually detectable development of a red coloration typified by an absorption peak at ca 530 nm. Conversely, in the presence of biothiols, reduction of Au(III) to Au(0) was prevented by entrapment of Au(III) within the biothiols via the thiol group. The solution thus remained colorless even after UV irradiation, which was used as an indicator of the presence of biothiols. Using this strategy, biothiols were very conveniently analyzed by monitoring color changes of the samples with the naked eye or a UV-vis spectrometer. The strategy based on this interesting phenomenon exhibited high selectivity toward biothiols over common amino acids and was successfully employed for reliable quantification of biothiols present in human plasma, demonstrating its great potential for clinical applications.

  15. Label-free colorimetric detection of biological thiols based on target-triggered inhibition of photoinduced formation of AuNPs.

    PubMed

    Jung, Ye Lim; Park, Jung Hun; Kim, Moon Il; Park, Hyun Gyu

    2016-02-01

    A label-free colorimetric method for the detection of biological thiols (biothiols) was developed. This method is based on prevention of the photoinduced reduction of auric ions (Au(III)) in the presence of amino acids (acting as a reducing agent) by biothiols; the photoinduced reduction is inhibited due to the strong interaction of the biothiols with Au(III). In this method, the sample was first incubated in an assay solution containing Au(III) and threonine; the sample solution was then exposed to 254 nm UV light. For samples without biothiols, this process led to the photoreduction of Au(III) followed by growth of gold nanoparticles accompanied by the visually detectable development of a red coloration typified by an absorption peak at ca 530 nm. Conversely, in the presence of biothiols, reduction of Au(III) to Au(0) was prevented by entrapment of Au(III) within the biothiols via the thiol group. The solution thus remained colorless even after UV irradiation, which was used as an indicator of the presence of biothiols. Using this strategy, biothiols were very conveniently analyzed by monitoring color changes of the samples with the naked eye or a UV-vis spectrometer. The strategy based on this interesting phenomenon exhibited high selectivity toward biothiols over common amino acids and was successfully employed for reliable quantification of biothiols present in human plasma, demonstrating its great potential for clinical applications. PMID:26671249

  16. Enhancement of Colorimetric Response of Enzymatic Reactions by Thermally Evaporated Plasmonic Thin Films: Application to Glial Fibrillary Acidic Protein

    PubMed Central

    Abel, Biebele; Kabir, Tabassum S.; Odukoya, Babatunde; Mohammed, Muzaffer; Aslan, Kadir

    2015-01-01

    We report the enhancement of the colorimetric response of horseradish peroxidase (HRP) and alkaline phosphatase (AP) in bioassays by thermally evaporated silver, gold, copper and nickel thin films. In this regard, a model bioassay based on biotin-avidin interactions was employed. Biotin groups and enzymes were introduced to all surfaces using a biotinylated linker molecule and avidin, respectively. The colorimetric response of HRP in the model bioassay carried out on the plasmonic thin films were up to 4.4-fold larger as compared to control samples (i.e., no plasmonic thin films), where the largest enhancement of colorimetric response was observed on silver thin films. The colorimetric response of AP on plasmonic thin films was found to be similar to those observed on control samples, which was attributed to the loss of enzymes from the surface during the bioassay steps. The extent of enzymes immobilized on to plasmonic thin films was found to affect the colorimetric response of the model bioassay. These findings allowed us to demonstrate the use of silver thin films for the detection of glial fibrillary acidic protein (GFAP), where the colorimetric response of the standard bioassays for GFAP was enhanced up to 67% as compared to bioassays on glass slides. PMID:25663850

  17. Label-free colorimetric sensor for ultrasensitive detection of heparin based on color quenching of gold nanorods by graphene oxide.

    PubMed

    Fu, Xiuli; Chen, Lingxin; Li, Jinhua; Lin, Meng; You, Huiyan; Wang, Wenhai

    2012-04-15

    A novel label-free colorimetric strategy was developed for ultrasensitive detection of heparin by using the super color quenching capacity of graphene oxide (GO). Hexadecyltrimethylammonium bromide (CTAB)-stabilized gold nanorods (AuNRs) could easily self-assembly onto the surface of GO through electrostatic interaction, resulting in decrease of the surface plasmon resonance (SPR) absorption and consequent color quenching change of the AuNRs from deep to light. Polycationic protamine was used as a medium for disturbing the electrostatic interaction between AuNRs and GO. The AuNRs were prevented from being adsorbed onto the surface of GO because of the stronger interaction between protamine and GO, showing a native color of the AuNRs. On the contrary, in the presence of heparin, which was more easily to combine with protamine, the AuNRs could self-assembly onto the surface of GO, resulting in the native color disappearing of AuNRs. As the concentration of heparin increased, the color of AuNRs would gradually fade until almost colorless. The amounts of self-assembly AuNRs were proportional to the concentration of heparin, and thereby the changes in the SPR absorption and color had been used to monitor heparin levels. Under optimized conditions, good linearity was obtained in a range of 0.02-0.28 μg/mL (R=0.9957), and a limit of detection was 5 ng/mL. The simultaneous possession of high sensitivity and selectivity, simplicity, rapidity, and visualization enabled this sensor to be potentially applicable for ultrasensitive and rapid on-site detection toward trace heparin. PMID:22387039

  18. Wrappers, Aspects, Quantification and Events

    NASA Technical Reports Server (NTRS)

    Filman, Robert E.

    2005-01-01

    Talk overview: Object infrastructure framework (OIF). A system development to simplify building distributed applications by allowing independent implementation of multiple concern. Essence and state of AOP. Trinity. Quantification over events. Current work on a generalized AOP technology.

  19. Chemically responsive nanoporous pigments: colorimetric sensor arrays and the identification of aliphatic amines.

    PubMed

    Bang, Jin Ho; Lim, Sung H; Park, Erwin; Suslick, Kenneth S

    2008-11-18

    A general method has been developed for the preparation of microspheres of nanoporous pigments, their formulation into chemically responsive pigment inks, and the printing of these inks as colorimetric sensor arrays. Using an ultrasonic-spray aerosol-gel synthesis from chemically responsive dyes and common silica precursors, 16 different nanoporous pigment microspheres have been prepared and characterized. New colorimetric sensor arrays have been created by printing inks of these chemically responsive pigments as primary sensor elements; these arrays have been successfully tested for the detection, identification, and quantitation of toxic aliphatic amines. Among 11 structurally similar amines, complete identification of each analyte without confusion was achieved using hierarchical cluster analysis (HCA). Furthermore, visual identification of ammonia gas was easily made at the IDLH (immediately dangerous to life or health), PEL (permissible exposure limits), and 0.1 PEL concentrations with high reproducibility. PMID:18950204

  20. Enzyme-free colorimetric detection systems based on the DNA strand displacement competition reaction

    NASA Astrophysics Data System (ADS)

    Zhang, Z.; Birkedal, V.; Gothelf, K. V.

    2016-05-01

    The strand displacement competition assay is based on the dynamic equilibrium of the competitive hybridization of two oligonucleotides (A and B) to a third oligonucleotide (S). In the presence of an analyte that binds to a specific affinity-moiety conjugated to strand B, the equilibrium shifts, which can be detected by a shift in the fluorescence resonance energy transfer signal between dyes attached to the DNA strands. In the present study we have integrated an ATP aptamer in the strand B and demonstrated the optical detection of ATP. Furthermore we explore a new readout method using a split G-quadruplex DNAzyme for colorimetric readout of the detection of streptavidin by the naked eye. Finally, we integrate the whole G-quadruplex DNAzyme system in a single DNA strand and show that it is applicable to colorimetric detection.

  1. Bio-functionalized silver nanoparticles for selective colorimetric sensing of toxic metal ions and antimicrobial studies.

    PubMed

    Vinod Kumar, V; Anbarasan, S; Christena, Lawrence Rene; SaiSubramanian, Nagarajan; Philip Anthony, Savarimuthu

    2014-08-14

    Hibiscus Sabdariffa (Gongura) plant extracts (leaves (HL) and stem (HS)) were used for the first time in the green synthesis of bio-functionalized silver nanoparticles (AgNPs). The bio-functionality of AgNPs has been successfully utilized for selective colorimetric sensing of potentially health and environmentally hazardous Hg(2+), Cd(2+) and Pb(2+) metal ions at ppm level in aqueous solution. Importantly, clearly distinguishable colour for all three metal ions was observed. The influence of extract preparation condition and pH were also explored on the formation of AgNPs. Both selectivity and sensitivity differed for AgNPs synthesized from different parts of the plant. Direct correlation between the stability of green synthesized AgNPs at different pH and its antibacterial effects has been established. The selective colorimetric sensing of toxic metal ions and antimicrobial effect of green synthesized AgNPs demonstrated the multifunctional applications of green nanotechnology. PMID:24717716

  2. Re-evaluation of colorimetric Cl- data from natural waters with DOC

    USGS Publications Warehouse

    Norton, S.A.; Handlet, M.J.; Kahl, J.S.; Peters, N.E.

    1996-01-01

    Colorimetric Cl- data from natural solutions that contain dissolved organic carbon (DOC) may be biased high. We evaluated aquatic Cl- concentrations in ecosystem compartments at the Bear Brook Watershed, Maine, and from lakes in Maine, using ion chromatography and colorimetry. DOC imparts a positive interference on colorimetric Cl- results proportional to DOC concentrations at approximately 0.8 ??eq Cl-/L per mg DOC/L. The interference is not a function of Cl- concentration. The resulting bias in concentrations of Cl- may be 50% or more of typical environmental values for Cl- in areas remote from atmospheric deposition of marine aerosols. Such biased data in the literature appear to have led to spurious conclusions about recycling of Cl- by forests, the usefulness of Cl- as a conservative tracer in watershed studies, and calculations of elemental budgets, ion balance, charge density of DOC, and dry deposition factors.

  3. Gold nanoparticles for the colorimetric and fluorescent detection of ions and small organic molecules

    NASA Astrophysics Data System (ADS)

    Liu, Dingbin; Wang, Zhuo; Jiang, Xingyu

    2011-04-01

    In recent years, gold nanoparticles (AuNPs) have drawn considerable research attention in the fields of catalysis, drug delivery, imaging, diagnostics, therapy and biosensors due to their unique optical and electronic properties. In this review, we summarized recent advances in the development of AuNP-based colorimetric and fluorescent assays for ions including cations (such as Hg2+, Cu2+, Pb2+, As3+, Ca2+, Al3+, etc) and anions (such as NO2-, CN-, PF6-, F-, I-, oxoanions), and small organic molecules (such as cysteine, homocysteine, trinitrotoluene, melamine and cocaine, ATP, glucose, dopamine and so forth). Many of these species adversely affect human health and the environment. Moreover, we paid particular attention to AuNP-based colorimetric and fluorescent assays in practical applications.

  4. Bio-functionalized silver nanoparticles for selective colorimetric sensing of toxic metal ions and antimicrobial studies

    NASA Astrophysics Data System (ADS)

    Vinod Kumar, V.; Anbarasan, S.; Christena, Lawrence Rene; SaiSubramanian, Nagarajan; Philip Anthony, Savarimuthu

    2014-08-01

    Hibiscus Sabdariffa (Gongura) plant extracts (leaves (HL) and stem (HS) were used for the first time in the green synthesis of bio-functionalized silver nanoparticles (AgNPs). The bio-functionality of AgNPs has been successfully utilized for selective colorimetric sensing of potentially health and environmentally hazardous Hg2+, Cd2+ and Pb2+ metal ions at ppm level in aqueous solution. Importantly, clearly distinguishable colour for all three metal ions was observed. The influence of extract preparation condition and pH were also explored on the formation of AgNPs. Both selectivity and sensitivity differed for AgNPs synthesized from different parts of the plant. Direct correlation between the stability of green synthesized AgNPs at different pH and its antibacterial effects has been established. The selective colorimetric sensing of toxic metal ions and antimicrobial effect of green synthesized AgNPs demonstrated the multifunctional applications of green nanotechnology.

  5. Design, Certification, and Deployment of the Colorimetric Water Quality Monitoring Kit (CWQMK)

    NASA Technical Reports Server (NTRS)

    Gazda, Daniel B.; Nolan, Daniel J.; Rutz, Jeff A.; Schultz, John R.; Siperko, Lorraine M.; Porter, Marc D.; Lipert, Robert J.; Flint, Stephanie M.; McCoy, J. Torin

    2010-01-01

    In August 2009, an experimental water quality monitoring kit based on Colorimetric Solid Phase Extraction (CSPE) technology was delivered to the International Space Station (ISS) aboard STS-128/17A. The kit, called the Colorimetric Water Quality Monitoring Kit (CWQMK), was flown and deployed as a Station Development Test Objective (SDTO) experiment on the ISS. The goal of the SDTO experiment is to evaluate the acceptability of CSPE technology for routine water quality monitoring on the ISS. This paper provides an overview of the SDTO experiment, as well as a detailed description of the CWQMK hardware and a summary of the testing and analysis conducted to certify the CWQMK for use on the ISS. The initial results obtained from the SDTO experiment are also reported and discussed in detail

  6. Direct Colorimetric Detection of a Receptor-Ligand Interaction by a Polymerized Bilayer Assembly

    NASA Astrophysics Data System (ADS)

    Charych, Deborah H.; Nagy, Jon O.; Spevak, Wayne; Bednarski, Mark D.

    1993-07-01

    Detection of receptor-ligand interactions is generally accomplished by indirect assays such as enzyme-linked immunosorbent assay. A direct colorimetric detection method based on a polydiacetylene bilayer assembled on glass microscope slides has been developed. The bilayer is composed of a self-assembled monolayer of octadecylsilane and a Langmuir-Blodgett monolayer of polydiacetylene. The polydiacetylene layer is functionalized with an analog of sialic acid, the receptor-specific ligand for the influenza virus hemagglutinin. The sialic acid ligand serves as a molecular recognition element and the conjugated polymer backbone signals binding at the surface by a chromatic transition. The color transition is readily visible to the naked eye as a blue to red color change and can be quantified by visible absorption spectroscopy. Direct colorimetric detection by polydiacetylene films offers new possibilities for diagnostic applications and screening for new drug candidates or binding ligands.

  7. Colorimetric and fluorometric detection of neomycin based on conjugated polydiacetylene supramolecules.

    PubMed

    Zhou, Guodong; Wang, Fang; Wang, Huilin; Kambam, Srinivasulu; Chen, Xiaoqiang

    2013-06-13

    Utilizing the colorimetric and fluorogenic changes, a system based on polydiacetylenes (PDAs) is developed for the detection of neomycin. The PDA supramolecules polymerized from the mixed liposome composed of N-(3-hydroxyphenyl)pentacosa-10,12-diynamide (PCDA-AP) and pentacosa-10,12-diynoic acid (PCDA) at an optimized ratio of 1:9 display a unique colorimetric change (blue to red) and fluorescent enhancement in the presence of neomycin. The detection limit for neomycin is estimated to be 2.55 × 10(-7) M by the fluorogenic method. The optical changes induced by neomycin can be attributed to the disruption of the hydrogen bonding between phenol and carboxylic acid from PCDA-AP and PCDA. PMID:23649672

  8. An overwhelmingly selective colorimetric sensor for Ag(+) using a simple modified polyacrylonitrile fiber.

    PubMed

    Xing, Xiaoli; Yang, Huixiao; Tao, Minli; Zhang, Wenqin

    2015-10-30

    A carboxymethyl-dithiocarbamate immobilized polyacrylonitrile fiber colorimetric sensor has been synthesized. This fiber sensor exhibits excellent selectivity and sensitivity for Ag(+) in aqueous solution with a remarkable color change from light pink to red-brown over a wide pH range of 2-12. The sensor responds selectively to Ag(+) in the presence of other ions, including Mg(2+), Al(3+), Ca(2+), Cr(3+), Mn(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+), Cd(2+), Hg(2+) and Pb(2+). The colorimetric sensor has an extremely fast response time (10s) and a low visual limit of detection (5.53×10(-12) mol/L). The fiber sensor also undergoes an obvious color change in the presence of Ag(+) solutions containing EDTA, NaCl or NaBr. Density functional theory optimization reveals that the sensor and Ag(+) interact via a seven-membered ring complexation mechanism. PMID:25967097

  9. Colorimetric evaluation of iPhone apps for colour vision tests based on the Ishihara test.

    PubMed

    Dain, Stephen J; AlMerdef, Ali

    2016-05-01

    Given the versatility of smart phone displays, it was inevitable that applications (apps) providing colour vision testing would appear as an option. In this study, the colorimetric characteristics of five available iPhone apps for colour vision testing are assessed as a prequel to possible clinical evaluation. The colours of the displays produced by the apps are assessed with reference to the colours of a printed Ishihara test. The visual task is assessed on the basis of the colour differences and the alignment to the dichromatic confusion lines. The apps vary in quality and while some are colorimetrically acceptable, there are also some problems with their construction in making them a clinically useful app rather than curiosity driven self-testing. There is no reason why, in principle, a suitable test cannot be designed for smart phones. PMID:27146711

  10. A real-time colorimetric and ratiometric fluorescent probe for sulfite.

    PubMed

    Wu, Ming-Yu; He, Ting; Li, Kun; Wu, Ming-Bo; Huang, Zheng; Yu, Xiao-Qi

    2013-05-21

    A real-time colorimetric and ratiometric fluorescent probe based on modulating the intramolecular charge transfer (ICT) of the coumarin platform for selective detection of sulfite is presented. This reaction based probe utilized the Michael addition to the dicyano-vinyl group with the detection limit of 5.8 × 10(-5) M. The probe displayed a high selectivity for sulfite over other anions and reactive sulfur especially for biothiols including cysteine (Cys), homocysteine (Hcy) and glutathione (GSH), with about 100 nm blue shift and more than 230 times intensity ratios change of the emission spectrum. Meanwhile, it could be easily observed that the probe for sulfite changes from red to pale yellow by the naked eye, and from red to blue under UV lamp immediately after the sulfite is added. To the best of our knowledge, it is the fastest response probe for sulfite ever reported, which could give a colorimetric and ratiometric fluorescent response instantly. PMID:23563108

  11. G-quadruplex-generating polymerase chain reaction for visual colorimetric detection of amplicons.

    PubMed

    Bhadra, Sanchita; Codrea, Vlad; Ellington, Andrew D

    2014-01-15

    We have developed a self-reporting polymerase chain reaction (PCR) system for visual colorimetric gene detection and distinction of single nucleotide polymorphisms (SNPs). Amplification is performed using target-specific primers modified with a 5'-end tail that is complementary to a G-quadruplex deoxyribozyme-forming sequence. At end-point, G-quadruplexes are forced to fold from PCR-generated duplex DNA and then are used to colorimetrically report the successful occurrence of PCR by assaying their peroxidase activity using a chromogenic substrate. Furthermore, primer design considerations for the G-quadruplex-generating PCR system have allowed us to visually distinguish SNPs associated with Mycobacterium tuberculosis drug resistance alleles. PMID:24135653

  12. Direct colorimetric detection of a receptor-ligand interaction by a polymerized bilayer assembly

    SciTech Connect

    Charych, D.H.; Nagy, J.O.; Bednarski, M.D. ); Spevak, W. )

    1993-07-30

    Detection of receptor-ligand interactions is generally accomplished by indirect assays such as enzyme-linked immunosorbent assay. A direct colorimetric detection method based on a polydiacetylene bilayer assembled on glass microscope slides has been developed. The bilayer is composed of a self-assembled monolayer of octadecylsilane and a Langmuir-Blodgett monolayer of polydiacetylene. The polydiacetylene layer is functionalized with an analog of sialic acid, the receptor-specific ligand for the influenza virus hemagglutinin. The sialic acid ligand serves as a molecular recognition element and the conjugated polymer backbone signals binding at the surface by a chromatic transition. The color transition is readily visible to the naked eye as a blue to red color change and can be quantified by visible absorption spectroscopy. Direct colorimetric detection by polydiacetylene films offers new possibilities for diagnostic applications and screening for new drug candidates or binding ligands.

  13. Colorimetric detection of hazardous gases using a remotely operated capturing and processing system.

    PubMed

    Montes-Robles, Roberto; Moragues, María Esperanza; Vivancos, José-Luis; Ibáñez, Javier; Fraile, Rubén; Martínez-Máñez, Ramón; García-Breijo, Eduardo

    2015-11-01

    This paper presents an electronic system for the automatic detection of hazardous gases. The proposed system implements colorimetric sensing algorithms, thus providing a low-cost solution to the problem of gas sensing. It is remotely operated and it performs the tasks of image capturing and processing, hence obtaining colour measurements in RGB (Red-Green-Blue) space that are subsequently sent to a remote operator via the internet. A prototype of the system has been built to test its performance. Specifically, experiments have been carried out aimed at the detection of CO, CO2, NO, NO2, SO2 and formaldehyde at diverse concentrations by using a chromogenic array composed by 13 active and 2 inert compounds. Statistical analyses of the results reveal a good performance of the electronic system and the feasibility of remote hazardous gas detection using colorimetric sensor arrays. PMID:26434416

  14. Nitrogen quantification with SNMS

    NASA Astrophysics Data System (ADS)

    Goschnick, J.; Natzeck, C.; Sommer, M.

    1999-04-01

    Plasma-based secondary neutral mass spectrometry (plasma SNMS) is a powerful analytical method for determining the elemental concentrations of almost any kind of material at low cost by using a cheap quadrupole mass filter. However, a quadrupole-based mass spectrometer is limited to nominal mass resolution. Atomic signals are sometimes superimposed by molecular signals (2 or 3 atomic clusters such as CH +, CH 2+ or metal oxide clusters) and/or intensities of double-charged species. Especially in the case of nitrogen several interferences can impede the quantification. This article reports on methods to recognize and deconvolute superpositions of N + with CH 2+, Li 2+, and Si 2+ at mass 14 D (Debye) occurring during analysis of organic and inorganic substances. The recognition is based on the signal pattern of N +, Li +, CH +, and Si +. The latter serve as indicators for a probable interference of molecular or double-charged species with N on mass 14 D. The subsequent deconvolution use different shapes of atomic and cluster kinetic energy distributions (kEDs) to determine the quantities of the intensity components by a linear fit of N + and non-atomic kEDs obtained from several organic and inorganic standards into the measured kED. The atomic intensity fraction yields a much better nitrogen concentration than the total intensity of mass 14 D after correction.

  15. Quantification of human responses

    NASA Technical Reports Server (NTRS)

    Steinlage, R. C.; Gantner, T. E.; Lim, P. Y. W.

    1992-01-01

    Human perception is a complex phenomenon which is difficult to quantify with instruments. For this reason, large panels of people are often used to elicit and aggregate subjective judgments. Print quality, taste, smell, sound quality of a stereo system, softness, and grading Olympic divers and skaters are some examples of situations where subjective measurements or judgments are paramount. We usually express what is in our mind through language as a medium but languages are limited in available choices of vocabularies, and as a result, our verbalizations are only approximate expressions of what we really have in mind. For lack of better methods to quantify subjective judgments, it is customary to set up a numerical scale such as 1, 2, 3, 4, 5 or 1, 2, 3, ..., 9, 10 for characterizing human responses and subjective judgments with no valid justification except that these scales are easy to understand and convenient to use. But these numerical scales are arbitrary simplifications of the complex human mind; the human mind is not restricted to such simple numerical variations. In fact, human responses and subjective judgments are psychophysical phenomena that are fuzzy entities and therefore difficult to handle by conventional mathematics and probability theory. The fuzzy mathematical approach provides a more realistic insight into understanding and quantifying human responses. This paper presents a method for quantifying human responses and subjective judgments without assuming a pattern of linear or numerical variation for human responses. In particular, quantification and evaluation of linguistic judgments was investigated.

  16. Colorimetric recognition of different enzymology-concerning transition metals based on a hybrid cluster complex.

    PubMed

    Li, Shun-Hua; Yuan, Wen-Tao; Zheng, Hong; Xu, Jin-Gou

    2004-07-01

    A hybrid cluster complex, formed by chelating a chromogenic ligand to a [2Fe-2S] cluster, sensitively exhibited differential colorimetric responses towards Hg2+, Cd2+, Cr3+, Pb2, Sn2+, Cu2+, Zn2+, Fe3+ and Co2+ in water at physiological pH. Speciation of some of these metal elements, such as Cr(III) and Sn(IV), was also studied by UV/Vis absorption. PMID:15293390

  17. A simple ratiometric and colorimetric chemosensor for the selective detection of fluoride in DMSO buffered solution

    NASA Astrophysics Data System (ADS)

    Niu, Hu; Shu, Qinghai; Jin, Shaohua; Li, Bingjun; Zhu, Jiaping; Li, Lijie; Chen, Shusen

    2016-01-01

    A derivative of squaramide (cyclobuta[b]quinoxaline-1, 2(3H, 8H)-dione) has been synthesized for the ratiometric and colorimetric sensing of F- in aqueous solution in competitive fashion. With F-, probe 1 showed a highly selective naked-eye detectable color change along with a characteristic UV-Vis absorbance over other tested ions, which probably originates from the deprotonation occurred between 1 and F-, as proved by the 1H NMR titration experiments and DFT calculations.

  18. Magnetic bead-based reverse colorimetric immunoassay strategy for sensing biomolecules.

    PubMed

    Gao, Zhuangqiang; Xu, Mingdi; Hou, Li; Chen, Guonan; Tang, Dianping

    2013-07-16

    A novel reverse colorimetric immunoassay (RCIA) strategy was for the first time designed and utilized for sensitive detection of low-abundance protein (prostate-specific antigen, PSA, used in this case) in biological fluids by coupling highly catalytic efficient catalase with magnetic bead-based peroxidase mimics. To construct such a RCIA system, two nanostructures including magnetic beads and gold nanoparticles were first synthesized and functionalized with anti-PSA capture antibody and catalase/anti-PSA detection antibody, respectively. Thereafter, a specific sandwich-type immunoassay format was employed for determination of PSA by using functional gold nanoparticles as enzymatic bioreactors and anti-PSA-conjugated magnetic beads as a colorimetric developer. The carried catalase, followed by the sandwiched immunocomplex, partially consumed the added hydrogen peroxide in the detection solution, which slowed down the catalytic efficiency of magnetic bead-based peroxidase mimics toward TMB/H2O2, thereby weakening the visible color and decreasing the colorimetric density. Different from conventional colorimetric immunoassay, the RCIA method determined the residual hydrogen peroxide in the substrate after consumption. Under the optimal conditions, the developed RCIA exhibited a wide dynamic range of 0.05-20 ng mL(-1) toward PSA with a detection limit of 0.03 ng mL(-1) at the 3Sblank level. Intra- and interassay coefficients of variation were below 6.1% and 9.3%, respectively. Additionally, the methodology was further validated for the analysis of 12 PSA clinical serum specimens, giving results in good accordance with those obtained by the commercially available enzyme-linked immunosorbent assay (ELISA) method. PMID:23806145

  19. A near infrared colorimetric and fluorometric probe for organophosphorus nerve agent mimics by intramolecular amidation.

    PubMed

    Hu, Xiao-Xiao; Su, Yue-Ting; Ma, Yun-Wei; Zhan, Xin-Qi; Zheng, Hong; Jiang, Yun-Bao

    2015-10-21

    A near infrared probe for sensitive colorimetric and fluorimetric detection of nerve agent mimics, DCP and DCNP, was reported based on the activation of a carboxylic acid group by the mimics to conduct an intramolecular amidation reaction in the heptamethine chromophore, where its absorption or excitation maximum wavelength could be greatly red-shifted by attenuating the electron-donating ability of the amine group in the bridgehead site of heptamethine cyanine. PMID:26323249

  20. A colorimetric and fluorescent probe for fluoride ions based on 6-acetyl-2-naphthol.

    PubMed

    Hou, Peng; Chen, Song; Song, Xiangzhi

    2014-08-01

    A colorimetric and turn-on fluorescent probe for fluoride ions, tert-butyldimethylsilane 6-acetyl-2-naphtholate, was readily synthesized from 6-acetyl-2-naphthol and tert-butyldimethylchlorosilane (TBSCl). The probe exhibits high sensitivity and good selectivity for fluoride ions in acetonitrile. The inherent mechanism involves the cleavage of the Si-O bond in the probe, which induced yellow color formation and prominent fluorescence enhancement. PMID:23881573

  1. Direct visualization of lead corona and its nanomolar colorimetric detection using anisotropic gold nanoparticles.

    PubMed

    Dwivedi, Charu; Chaudhary, Abhishek; Gupta, Abhishek; Nandi, Chayan K

    2015-03-11

    The study presents dithiothreitol (DTT) functionalized anisotropic gold nanoparticles (GNP) based colorimetric sensor for detection of toxic lead ions in water. Our results demonstrate the selectivity and sensitivity of the developed sensor over various heavy metal ions with detection limit of ∼9 nM. The mechanism of sensing is explained on the basis of unique corona formation around the DTT functionalized anisotropic GNP. PMID:25719820

  2. Chromatic distortion during angioscopy: assessment and correction by quantitative colorimetric angioscopic analysis.

    PubMed

    Lehmann, K G; Oomen, J A; Slager, C J; deFeyter, P J; Serruys, P W

    1998-10-01

    Angioscopy represents a diagnostic tool with the unique ability of assessing the true color of intravascular structures. Current angioscopic interpretation is entirely subjective, however, and the visual interpretation of color has been shown to be marginal at best. The quantitative colorimetric angioscopic analysis system permits the full characterization of angioscopic color using two parameters (C1 and C2), derived from a custom color coordinate system, that are independent of illuminating light intensity. Measurement variability was found to be low (coefficient of variation = 0.06-0.64%), and relatively stable colorimetric values were obtained even at the extremes of illumination power. Variability between different angioscopic catheters was good (maximum difference for C1, 0.022; for C2, 0.015). Catheter flexion did not significantly distort color transmission. Although the fiber optic illumination bundle was found to impart a slight yellow tint to objects in view (deltaC1 = 0.020, deltaC2 = 0.024, P < 0.0001) and the imaging bundle in isolation imparted a slight red tint (deltaC1 = 0.043, deltaC2 = -0.027, P < 0.0001), both of these artifacts could be corrected by proper white balancing. Finally, evaluation of regional chromatic characteristics revealed a radially symmetric and progressive blue shift in measured color when moving from the periphery to the center of an angioscopic image. An algorithm was developed that could automatically correct 93.0-94.3% of this error and provide accurate colorimetric measurements independent of spatial location within the angioscopic field. In summary, quantitative colorimetric angioscopic analysis provides objective and highly reproducible measurements of angioscopic color. This technique can correct for important chromatic distortions present in modern angioscopic systems. It can also help overcome current limitations in angioscopy research and clinical use imposed by the reliance on visual perception of color. PMID

  3. Use of photopolymerization for the rapid and cost-effective identification of Shiga toxin-producing Escherichia coli on DNA microarrays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Shiga toxin-producing Escherichia coli O157:H7 is a leading cause of foodborne illness worldwide. To evaluate better methods to rapidly detect and genotype E. coli O157 virulent strains, the present study explored the use of photopolymerization, a colorimetric and photoinduced signal amplification d...

  4. Colorimetric monitoring of formaldehyde in indoor environment using built-in camera on mobile phone.

    PubMed

    Sekine, Yoshika; Katori, Risa; Tsuda, Yuko; Kitahara, Takio

    2016-01-01

    A simple monitoring system of indoor air pollution is proposed by integrating a novel colorimetric detector of formaldehyde (HCHO) and a function of a built-in camera on mobile phone. The colorimetric detector employs a solid phase colorimetric reagent made from 4-amino-3-hydrazino-5-mercapto-1,2,4-triazole, ZnO, KIO4 and agar, and changes colour from white to purple by exposure to HCHO gas. The degree of colour changes expressed in Red, Green and Blue model model responded to the HCHO concentration levels both in air and from building materials. Limit of quantitation of the detector with 24 h-exposure resulted in 0.011 mg/m(3) of air concentration which meets a requirement of methodology to detect indoor air quality guideline level of HCHO set by World Health Organization. The detector is also applicable to classify HCHO-emitting materials at least into Type 1, whose emission flux is greater than 120 μg/m(2)/h, and others. Then, variation of the acquired photo images was investigated by using various mobile phones and changing conditions of photography. As a result, the calibration of the measured colour intensity with a colour standard reduced the variation of the results and gave a significant output when the auto-focused images were taken under the condition of common indoor environment. PMID:26616679

  5. Label-free colorimetric aptasensor for sensitive detection of ochratoxin A utilizing hybridization chain reaction.

    PubMed

    Wang, Chengke; Dong, Xiaoya; Liu, Qian; Wang, Kun

    2015-02-20

    The combination of high selectivity of aptamer with the peroxidase-mimicking property of DNAzyme has presented considerable opportunities for designing colorimetric aptasensor for detection of ochratoxin A (OTA). The activities of both aptamer (as biorecognition element) and DNAzyme (as signal amplification element) are blocked via base pairing in the hairpin structure. Hybridization chain reaction (HCR) between two hairpin DNAs was employed to further improve the sensitivity of this method. The presence of OTA triggers the opening of the hairpin structure and the beginning of HCR, which results in the release of many DNAzyme, and generates enhanced colorimetric signals, which is correlated to the amounts of OTA with linear range between 0.01 to 0.32 nM, and the limit of detection is 0.01 nM under optimal conditions. OTA in yellow rice wine and wheat flour samples was also detected using this method. We demonstrate that a new colorimetric method for the detection of OTA has been established, which is simple, easy to conduct, label-free, sensitive, high throughput, and cost-saving. PMID:25682251

  6. Colorimetric cholesterol sensor based on peroxidase like activity of zinc oxide nanoparticles incorporated carbon nanotubes.

    PubMed

    Hayat, Akhtar; Haider, Waqar; Raza, Yousuf; Marty, Jean Louis

    2015-10-01

    A sensitive and selective colorimetric method based on the incorporation of zinc oxide nanoparticles (ZnO NPs) on the surface of carbon nanotubes (CNTs) was shown to posses synergistic peroxidase like activity for the detection of cholesterol. The proposed nanocomposite catalyzed the oxidation of 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) in the presence of hydrogen peroxide (H2O2) to produce a green colored product which can be monitored at 405 nm. H2O2 is the oxidative product of cholesterol in the presence of cholesterol oxidase. Therefore, the oxidation of cholesterol can be quantitatively related to the colorimetric response by combining these two reactions. Under the optimal experimental conditions, the colorimetric response was proportional to the concentration of cholesterol in the range of 0.5-500 nmol/L, with a detection limit of 0.2 nmol/L. The applicability of the proposed assays was demonstrated for the determination of cholesterol in milk powder samples with good recovery results. PMID:26078143

  7. The Use of Colorimetric Sensor Arrays to Discriminate between Pathogenic Bacteria

    PubMed Central

    Lonsdale, Claire L.; Taba, Brian; Queralto, Nuria; Lukaszewski, Roman A.; Martino, Raymond A.; Rhodes, Paul A.; Lim, Sung H.

    2013-01-01

    A colorimetric sensor array is a high-dimensional chemical sensor that is cheap, compact, disposable, robust, and easy to operate, making it a good candidate technology to detect pathogenic bacteria, especially potential bioterrorism agents like Yersinia pestis and Bacillus anthracis which feature on the Center for Disease Control and Prevention’s list of potential biothreats. Here, a colorimetric sensor array was used to continuously monitor the volatile metabolites released by bacteria in solid media culture in an Advisory Committee on Dangerous Pathogen Containment Level 3 laboratory. At inoculum concentrations as low as 8 colony-forming units per plate, 4 different bacterial species were identified with 100% accuracy using logistic regression to classify the kinetic profile of sensor responses to culture headspace gas. The sensor array was able to further discriminate between different strains of the same species, including 5 strains of Yersinia pestis and Bacillus anthracis. These preliminary results suggest that disposable colorimetric sensor arrays can be an effective, low-cost tool to identify pathogenic bacteria. PMID:23671629

  8. A cellulose-based bioassay for the colorimetric detection of pathogen DNA.

    PubMed

    Saikrishnan, Deepika; Goyal, Madhu; Rossiter, Sharon; Kukol, Andreas

    2014-12-01

    Cellulose-paper-based colorimetric bioassays may be used at the point of sampling without sophisticated equipment. This study reports the development of a colorimetric bioassay based on cellulose that can detect pathogen DNA. The detection was based on covalently attached single-stranded DNA probes and visual analysis. A cellulose surface functionalized with tosyl groups was prepared by the N,N-dimethylacetamide-lithium chloride method. Tosylation of cellulose was confirmed by scanning electron microscopy, Fourier transform infrared spectroscopy and elemental analysis. Sulfhydryl-modified oligonucleotide probes complementary to a segment of the DNA sequence IS6110 of Mycobacterium tuberculosis were covalently immobilized on the tosylated cellulose. On hybridization of biotin-labelled DNA oligonucleotides with these probes, a colorimetric signal was obtained with streptavidin-conjugated horseradish peroxidase catalysing the oxidation of tetramethylbenzamidine by H2O2. The colour intensity was significantly reduced when the bioassay was subjected to DNA oligonucleotide of randomized base composition. Initial experiments have shown a sensitivity of 0.1 μM. A high probe immobilization efficiency (more than 90 %) was observed with a detection limit of 0.1 μM, corresponding to an absolute amount of 10 pmol. The detection of M. tuberculosis DNA was demonstrated using this technique coupled with PCR for biotinylation of the DNA. This work shows the potential use of tosylated cellulose as the basis for point-of-sampling bioassays. PMID:25354892

  9. Thermodynamic study of colorimetric transitions in polydiacetylene vesicles induced by the solvent effect.

    PubMed

    Pires, Ana Clarissa S; Soares, Nilda de Fátima F; da Silva, Luis Henrique M; da Silva, Maria C Hespanhol; Mageste, Aparecida B; Soares, Rêmili F; Teixeira, Álvaro V N C; Andrade, Nélio J

    2010-10-28

    We report the synthesis of 10,12-pentacosadyinoic acid (PCDA) and PCDA + cholesterol (CHO) + sphingomyelin (SPH) vesicles dispersed in water and the determination of their colorimetric response induced by small amount of organic solvents. In the absence of solvent, PCDA and PCDA/CHO/SPH vesicles showed an intense blue color. The addition of CHCl(3), CH(2)Cl(2), and CCl(4) caused a colorimetric transition (CT) in both structures with the following efficiency: CHCl(3) > CH(2)Cl(2) ≅ CCl(4). However, CH(3)OH did not cause a blue-to-red transition. By microcalorimetric technique we also determined, for the first time, the enthalpy change associated with the CT process and the energy of interaction between solvent molecules and vesicle self-assembly. We observed that the chloride solvents induced a colorimetric transition, but the thermodynamic mechanism was different for each of them. CT induced by CHCl(3) was enthalpically driven, while that caused by CH(2)Cl(2) or CCl(4) was entropically driven. PMID:20883002

  10. Microfluidic chip-based silver nanoparticles aptasensor for colorimetric detection of thrombin.

    PubMed

    Zhao, Yaju; Liu, Xiaohui; Li, Jie; Qiang, Weibing; Sun, Liang; Li, Hui; Xu, Danke

    2016-04-01

    In this paper, a colorimetric silver nanoparticles aptasensor (aptamer-AgNPs) was developed for simple and straightforward detection of protein in microfluidic chip. Surface-functionalized microfluidic channels were employed as the capture platform. Then the mixture of target protein and aptamer-AgNPs were injected into the microfluidic channels for colorimetric detection. To demonstrate the performance of this detection platform, thrombin was chosen as a model target protein. Introduction of thrombin could form a sandwich-type complex involving immobilized AgNPs. The amount of aptamer-AgNPs on the complex augmented along with the increase of the thrombin concentration causing different color change that can be analyzed both by naked eyes and a flatbed scanner. This method is featured with low sample consumption, simple processes of microfluidic platform and straightforward colorimetric detection with aptamer-AgNPs. Thrombin at concentrations as low as 20pM can be detected using this aptasensor without signal amplification. This work demonstrated that it had good selectivity over other proteins and it could be a useful strategy to detect other targets with two affinity binding sites for ligands as well. PMID:26838384

  11. Double-stem Hairpin Probe and Ultrasensitive Colorimetric Detection of Cancer-related Nucleic Acids

    PubMed Central

    Xu, Jianguo; Li, Hongling; Wu, Zai-Sheng; Qian, Jun; Xue, Chang; Jia, Lee

    2016-01-01

    The development of a versatile biosensing platform to screen specific DNA sequences is still an essential issue of molecular biology research and clinic diagnosis of genetic disease. In this work, we for the first time reported a double-stem hairpin probe (DHP) that was simultaneously engineered to incorporate a DNAzyme, DNAzyme's complementary fragment and nicking enzyme recognition site. The important aspect of this hairpin probe is that, although it is designed to have a long ds DNA fragment, no intermolecular interaction occurs, circumventing the sticky-end pairing-determined disadvantages encountered by classic molecular beacon. For the DHP-based colorimetric sensing system, as a model analyte, cancer-related DNA sequence can trigger a cascade polymerization/nicking cycle on only one oligonucleotide probe. This led to the dramatic accumulation of G-quadruplexes directly responsible for colorimetric signal conversion without any loss. As a result, the target DNA is capable of being detected to 1 fM (six to eight orders of magnitude lower than that of catalytic molecular beacons) and point mutations are distinguished by the naked eye. The described DHP as a-proof-of-concept would not only promote the design of colorimetric biosensors but also open a good way to promote the diagnosis and treatment of genetic diseases. PMID:26909108

  12. Aptamer-based colorimetric sensing of acetamiprid in soil samples: sensitivity, selectivity and mechanism.

    PubMed

    Shi, Huijie; Zhao, Guohua; Liu, Meichuan; Fan, Lifang; Cao, Tongcheng

    2013-09-15

    A simple and selective aptamer-based colorimetric method was developed for highly sensitive detection of acetamiprid, taking advantages of the sensitive target-induced colour changes that arisen from the interparticle plasmon coupling during the aggregation of Au nanoparticles (NPs). The results showed that the established method could be applied to detect acetamiprid in the linear range between 75 nM to 7.5 μM, with a low detection limit of 5 nM. Meanwhile, by employing an "artificial antibody" acetamiprid-binding aptamer (ABA) as recognition element, highly selective and specific colorimetric visualization of acetamiprid was obtained. It indicated that pesticides which may coexist with acetamiprid could not interfere with the detection of acetamiprid even that had similar structure with acetamiprid, such as imidacloprid and chlorpyrifos. Mechanism study suggested that it could be attributed to the specific supramolecular interaction between ABA and acetamiprid, as well as the resulted target-binding event induced conformation changes of ABA from random coil to hairpin structure. The practical application of the colorimetric method was realized for detecting acetamiprid in real soil samples and monitoring its natural degradation process. PMID:23846126

  13. Double-stem Hairpin Probe and Ultrasensitive Colorimetric Detection of Cancer-related Nucleic Acids.

    PubMed

    Xu, Jianguo; Li, Hongling; Wu, Zai-Sheng; Qian, Jun; Xue, Chang; Jia, Lee

    2016-01-01

    The development of a versatile biosensing platform to screen specific DNA sequences is still an essential issue of molecular biology research and clinic diagnosis of genetic disease. In this work, we for the first time reported a double-stem hairpin probe (DHP) that was simultaneously engineered to incorporate a DNAzyme, DNAzyme's complementary fragment and nicking enzyme recognition site. The important aspect of this hairpin probe is that, although it is designed to have a long ds DNA fragment, no intermolecular interaction occurs, circumventing the sticky-end pairing-determined disadvantages encountered by classic molecular beacon. For the DHP-based colorimetric sensing system, as a model analyte, cancer-related DNA sequence can trigger a cascade polymerization/nicking cycle on only one oligonucleotide probe. This led to the dramatic accumulation of G-quadruplexes directly responsible for colorimetric signal conversion without any loss. As a result, the target DNA is capable of being detected to 1 fM (six to eight orders of magnitude lower than that of catalytic molecular beacons) and point mutations are distinguished by the naked eye. The described DHP as a-proof-of-concept would not only promote the design of colorimetric biosensors but also open a good way to promote the diagnosis and treatment of genetic diseases. PMID:26909108

  14. A colorimetric and fluorescent probe for detecting intracellular biothiols.

    PubMed

    Chen, Chunyang; Liu, Wei; Xu, Cong; Liu, Weisheng

    2016-11-15

    A new rapid and highly sensitive coumarin-based probe (probe 1) has been designed and synthesized for detecting intracellular thiols. Probe 1 was prepared by a 4-step procedure as a latent fluorescence probe to achieve high sensitivity and fluorescence turn-on response toward cysteine and homocysteine over GSH and other various natural amino acids under physiological conditions. Owing to specific cyclization between thiols and aldehyde group, probe 1 displayed a highly selectivity toward cysteine and homocysteine. Above all, probe 1 was successfully used for fluorescence imaging of biothiols in Hela cells, and quantitative determination had been achieved within a certain range. Then specific fluorescence imaging of mice organ tissues was obtained for proving the permeability of probe 1. Simultaneously, the viability was measured to be more than 80%, which shows probe 1 can be a rapid and biocompatible probe for biothiols in cells. Furthermore, the measurement of thiols detection in 5 kinds of animal serum showed that probe 1 can be used in determination of biothiols in blood. PMID:27155115

  15. Quantification of Glutathione in Caenorhabditis elegans

    PubMed Central

    Caito, Samuel W.; Aschner, Michael

    2015-01-01

    Glutathione (GSH) is the most abundant intracellular thiol with diverse functions from redox signaling, xenobiotic detoxification, and apoptosis. The quantification of GSH is an important measure for redox capacity and oxidative stress. This protocol quantifies total GSH from Caenorhabditis elegans, an emerging model organism for toxicology studies. GSH is measured using the 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB) cycling method originally created for cell and tissue samples but optimized for whole worm extracts. DTNB reacts with GSH to from a 5′-thio-2-nitrobenzoic acid (TNB) chromophore with maximum absorbance of 412 nm. This method is both rapid and sensitive, making it ideal for studies involving a large number of transgenic nematode strains. PMID:26309452

  16. Label-free colorimetric biosensing of copper(II) ions with unimolecular self-cleaving deoxyribozymes and unmodified gold nanoparticle probes

    NASA Astrophysics Data System (ADS)

    Wang, Yong; Yang, Fan; Yang, Xiurong

    2010-05-01

    Using unimolecular copper(II)-dependent self-cleaving deoxyribozymes (DNAzymes), a label-free colorimetric biosensor for copper(II) ions (Cu2 + ) has been developed based on the sequence-length-dependent adsorption of single-stranded deoxyribonucleic acid (ssDNA) on unmodified gold nanoparticles (AuNPs). In the presence of Cu2 + , the Cu2 + -dependent DNAzyme could be self-cleaved into short ssDNA fragments. The cleaved short ssDNA could adsorb rapidly onto the surface of the AuNPs. This enhanced the stability of the AuNPs against salt-induced aggregation, and thus the solution color remained red. In the absence of Cu2 + , however, uncleaved long ssDNA adsorbed relatively slowly onto the AuNPs and upon the addition of salt, the electrostatic repulsion between the AuNPs was screened, resulting in aggregation of the AuNPs which produced a red-to-blue color change. Thus, Cu2 + detection could be realized by monitoring the color change of the AuNPs. The calibration curve showed that the absorption ratio values at 520 and 620 nm increased linearly over the Cu2 + concentration range of 0.625-15 µM, with a limit of detection of 290 nM. The other environmentally relevant metal ions did not interfere with the determination of Cu2 + . Subsequently, the assay was employed to determine Cu2 + in several water samples, and the results were satisfactory. It is expected that the present colorimetric strategy will be possibly extended to the detection of cofactors of other in vitro-selected unimolecular self-cleaving DNAzymes, such as amino acids, nucleic acids, metal ions and small organic molecules.

  17. Highly selective and sensitive paper-based colorimetric sensor using thiosulfate catalytic etching of silver nanoplates for trace determination of copper ions.

    PubMed

    Chaiyo, Sudkate; Siangproh, Weena; Apilux, Amara; Chailapakul, Orawon

    2015-03-25

    A novel, highly selective and sensitive paper-based colorimetric sensor for trace determination of copper (Cu(2+)) ions was developed. The measurement is based on the catalytic etching of silver nanoplates (AgNPls) by thiosulfate (S2O3(2-)). Upon the addition of Cu(2+) to the ammonium buffer at pH 11, the absorption peak intensity of AuNPls/S2O3(2-) at 522 nm decreased and the pinkish violet AuNPls became clear in color as visible to the naked eye. This assay provides highly sensitive and selective detection of Cu(2+) over other metal ions (K(+), Cr(3+), Cd(2+), Zn(2+), As(3+), Mn(2+), Co(2+), Pb(2+), Al(3+), Ni(2+), Fe(3+), Mg(2+), Hg(2+) and Bi(3+)). A paper-based colorimetric sensor was then developed for the simple and rapid determination of Cu(2+) using the catalytic etching of AgNPls. Under optimized conditions, the modified AgNPls coated at the test zone of the devices immediately changes in color in the presence of Cu(2+). The limit of detection (LOD) was found to be 1.0 ng mL(-1) by visual detection. For semi-quantitative measurement with image processing, the method detected Cu(2+) in the range of 0.5-200 ng mL(-1)(R(2)=0.9974) with an LOD of 0.3 ng mL(-1). The proposed method was successfully applied to detect Cu(2+) in the wide range of real samples including water, food, and blood. The results were in good agreement according to a paired t-test with results from inductively coupled plasma-optical emission spectrometry (ICP-OES). PMID:25732695

  18. Photonic nanosensor for colorimetric detection of metal ions.

    PubMed

    Yetisen, Ali K; Montelongo, Yunuen; Qasim, Malik M; Butt, Haider; Wilkinson, Timothy D; Monteiro, Michael J; Yun, Seok Hyun

    2015-01-01

    The real-time sensing of metal ions at point of care requires integrated sensors with low energy and sample consumption, reversibility, and rapid recovery. Here, we report a photonic nanosensor that reversibly and quantitatively reports on variation in the concentrations of Pb(2+) and Cu(2+) ions in aqueous solutions (<500 μL) in the visible region of the spectrum (λ(max) ≈ 400-700 nm). A single 6 ns laser pulse (λ = 532 nm) was used to pattern an ∼10 μm thick photosensitive recording medium. This formed periodic AgBr nanocrystal (ø ∼ 5-20 nm) concentrated regions, which produced Bragg diffraction upon illumination with a white light source. The sensor functionalized with 8-hydroxyquinoline allowed sensing through inducing Donnan osmotic pressure and tuning its lattice spacing. The sensor quantitatively measured Pb(2+) and Cu(2+) ion concentrations within the dynamic range of 0.1-10.0 mM with limits of detection of 11.4 and 18.6 μM in under 10 min. The sensor could be reset in 3 min and was reused at least 100 times without compromising its accuracy. The plasmonic nanosensor represents a simple and label-free analytical platform with potential scalability for applications in medical diagnostics and environmental monitoring. PMID:25710792

  19. A label-free colorimetric detection of lead ions by controlling the ligand shells of gold nanoparticles.

    PubMed

    Hung, Yu-Lun; Hsiung, Tung-Ming; Chen, Yi-You; Huang, Chih-Ching

    2010-07-15

    We have developed a simple, colorimetric and label-free gold nanoparticle (Au NP)-based probe for the detection of Pb(2+) ions in aqueous solution, operating on the principle that Pb(2+) ions change the ligand shell of thiosulfate (S(2)O(3)(2-))-passivated Au NPs. Au NPs reacted with S(2)O(3)(2-) ions in solution to form Au(+).S(2)O(3)(2-) ligand shells on the Au NP surfaces, thereby inhibiting the access of 4-mercaptobutanol (4-MB). Surface-assisted laser desorption/ionization time-of-flight ionization mass spectrometry (SALDI-TOF MS) and inductively coupled plasma mass spectrometry (ICP-MS) measurements revealed that PbAu alloys formed on the surfaces of the Au NPs in the presence of Pb(2+) ions; these alloys weakened the stability of the Au(+).S(2)O(3)(2-) ligand shells, enhancing the access of 4-MB to the Au NP surfaces and, therefore, inducing their aggregation. As a result, the surface plasmon resonance (SPR) absorption of the Au NPs red-shifted and broadened, allowing quantitation of the Pb(2+) ions in the aqueous solution. This 4-MB/S(2)O(3)(2-)-Au NP probe is highly sensitive (linear detection range: 0.5-10 nM) and selective (by at least 100-fold over other metal ions) toward Pb(2+) ions. This cost-effective sensing system allows the rapid and simple determination of the concentrations of Pb(2+) ions in real samples (in this case, river water, Montana soil and urine samples). PMID:20602929

  20. A colorimetric nitrite detection system with excellent selectivity and high sensitivity based on Ag@Au nanoparticles.

    PubMed

    Li, Tianhua; Li, Yonglong; Zhang, Yujie; Dong, Chen; Shen, Zheyu; Wu, Aiguo

    2015-02-21

    Excessive uptake of NO2(-) is detrimental to human health, but the currently available methods used to sensitively detect this ion in the environment are cumbersome and expensive. In this study, we developed an improved NO2(-) detection system based on a redox etching strategy of CTAB-stabilized Ag-Au core-shell nanoparticles (Ag@AuNPs). The detection mechanism was verified by UV-Vis spectroscopy, TEM and XPS. The detection system produces a color change from purple to colorless in response to an increase of NO2(-) concentration. The selectivity of detection of NO2(-), both with the unaided eye and by measurement of UV-Vis spectra, is excellent in relation to other ions, including Cu(2+), Co(2+), Ni(2+), Cr(3+), Al(3+), Pb(2+), Cd(2+), Ca(2+), Ba(2+), Zn(2+), Mn(2+), Mg(2+), Fe(3+), Hg(2+), Ag(+), K(+), F(-), PO4(3-), C2O4(2-), SO3(2-), CO3(2-), SO4(2-), NO3(-) and CH3-COO(-) (Ac(-)). The limit of detection (LOD) for NO2(-) is 1.0 μM by eye and 0.1 μM by UV-Vis spectroscopy. The LOD by eye is lower than the lowest previously reported value (4.0 μM). There is a good linear relationship between A/A0 and the concentration of NO2(-) from 1.0 to 20.0 μM NO2(-), which permits a quantitative assay. The applicability of our detection system was also verified by analysis of NO2(-) in tap water and lake water. The results demonstrate that our Ag@AuNP-based detection system can be used for the rapid colorimetric detection of NO2(-) in complex environmental samples, with excellent selectivity and high sensitivity. PMID:25564225

  1. Cost-effective and sensitive colorimetric immunosensing using an iron oxide-to-Prussian blue nanoparticle conversion strategy.

    PubMed

    Fu, Guanglei; Sanjay, Sharma T; Li, XiuJun

    2016-06-21

    The development of new sensitive, cost-effective and user-friendly colorimetric bioassays is in increasing demand to meet the requirement of modern clinical diagnostics and field detection. Herein, a novel iron oxide-to-Prussian blue (PB) nanoparticle (NP) conversion strategy was developed and applied to sensitive colorimetric immunosensing of cancer biomarkers. In a typical sandwich-type immunosensing system, the captured spherical antibody-conjugated iron oxide NPs were transformed into cubic PB NPs, which exhibited a highly visible blue color with high molar extinction coefficients. Hence, a new colorimetric immunosensing strategy was developed as a result of this low cost and simple transformation process. Without the aid of any complex nanoparticle stabilizing ligands and signal amplification processes, prostate-specific antigen as a model analyte can be detected at a concentration as low as 1.0 ng mL(-1) by the naked eye with good reliability for detection of real human serum samples. This is the first attempt to develop and apply the iron oxide-to-PB NP colorimetric conversion strategy for immunosensing, and shows great promise for the development of new sensitive, cost-effective and user-friendly colorimetric bioassays in various bioanalytical applications, especially in low-resource settings. PMID:27140740

  2. MAMA Software Features: Quantification Verification Documentation-1

    SciTech Connect

    Ruggiero, Christy E.; Porter, Reid B.

    2014-05-21

    This document reviews the verification of the basic shape quantification attributes in the MAMA software against hand calculations in order to show that the calculations are implemented mathematically correctly and give the expected quantification results.

  3. 4-(8-Quinolyl)amino-7-nitro-2,1,3-benzoxadiazole as a new selective and sensitive fluorescent and colorimetric pH probe with dual-responsive ranges in aqueous solutions

    NASA Astrophysics Data System (ADS)

    Li, Xutian; Zhang, Min; Liang, Haipeng; Huang, Zhaowei; Tang, Jiang; Chen, Zhi; Yang, Liting; Ma, Li-Jun; Wang, Yuhai; Xu, Baiping

    2016-01-01

    Fluorescent and colorimetric pH probe possess many advantages including rapid response time, nondestructive testing, and excellent pH sensitivity. However, they usually cannot be utilized simultaneously in both acidic and basic pH ranges. In this study, a new selective and sensitive fluorescent and colorimetric pH probe, 4-(8- quinolyl)amino-7-nitro-2,1,3-benzoxadiazole (1), was designated and synthesized. The optical probe exhibited dual-responsive pH ranges to both acidic and basic aqueous solutions. When the solution pH was gradually increased from 8.5 to 13.3, the absorption spectra of 1 showed an obvious hyperchromicity, accompanied with a red shift of the absorption band at 340 nm, a blue shift of the absorption band at 482 nm, and a distinct color change from orange to violet pink to yellow. Within the pH range from 2.2 to 0.2, the fluorescent spectra of 1 showed a "turn-on" response signal to solution pH. In order to understand the response mechanism of the probe to solution pH, the probe molecule was split into two parts, 8-aminoquinoline (2) and 4-amino-7- nitro-benzofurazan (3). UV-vis absorption and fluorescent experiments of 2 and 3 indicated that both are sensitive optical pH probes. Furthermore, the NMR experiment of 1 was explored in basic and acidic conditions. The results indicated that the colorimetric responses of 1 to pH under basic condition should be attributed to the deprotonation of the imino group on the quinolyl ring, and the fluorescent recognition of 1 to pH under acidic condition was probably due to the protonation of the nitrogen atoms from the benzofurazan and quinolyl rings.

  4. Rapid Prototyping

    NASA Technical Reports Server (NTRS)

    1999-01-01

    Javelin, a Lone Peak Engineering Inc. Company has introduced the SteamRoller(TM) System as a commercial product. The system was designed by Javelin during a Phase II NASA funded small commercial product. The purpose of the invention was to allow automated-feed of flexible ceramic tapes to the Laminated Object Manufacturing rapid prototyping equipment. The ceramic material that Javelin was working with during the Phase II project is silicon nitride. This engineered ceramic material is of interest for space-based component.

  5. 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.

  6. Inkjet-printed paper-based colorimetric sensor array for the discrimination of volatile primary amines.

    PubMed

    Soga, Tamaki; Jimbo, Yusuke; Suzuki, Koji; Citterio, Daniel

    2013-10-01

    This paper describes a colorimetric sensor array for the discrimination of volatile amines. Analyte discrimination is achieved by combining two functional elements: (1) a "chemical class-selective" single chromogenic sensing dye with selectivity for amines in general, encapsulated into (2) polymer nanoparticles with different polarities. The resulting array has the ability to distinguish one closely related amine from another, relying on a polarity-based approach. In order to achieve reproducible, cost efficient, and flexible sensor array fabrication with the potential for mass production, inkjet-printing technology combined with standard copy paper as a sensor substrate is applied. Printing of 6 types of inks, which are prepared by mixing two dye encapsulating nanoparticles of different polarity in different mixture ratios, results in a colorimetric sensor array with a polarity gradient. Seven primary amines with increasing alkyl chain lengths have been selected to demonstrate the performance of the sensor array. The RGB color differences (ΔR, ΔG, ΔB) of the sensor array spots before and after gas exposure were analyzed by principal component analysis (PCA) and agglomerative hierarchical clustering (AHC) analysis. Under the selected measurement conditions, results of PCA and AHC analysis indicated high discrimination ability with high reproducibility of the sensor array down to amine concentrations of 50 ppm. The discrimination ability was maintained at relative humidities between 10% and 80%. Furthermore, the sensor array showed no significant response to common volatile organic compounds, confirming the high selectivity toward amines. This is, to the best of our knowledge, the first report of a colorimetric sensor array with selectivity for a specific chemical class of analytes and the ability to discriminate compounds of the same class, which is obtained by simply mixing two types of single dye-encapsulating polymer nanoparticles. PMID:24044503

  7. Urinary Colorimetric Sensor Array and Algorithm to Distinguish Kawasaki Disease from Other Febrile Illnesses

    PubMed Central

    Jin, Bo; Deng, Xiaohong; Hu, Guang; Liu, Xiaodan; Zhang, Jie; Jin, Hua; Huang, Min; Kanegaye, John T.; Tremoulet, Adriana H.; Burns, Jane C.; Wu, Jianmin; Cohen, Harvey J.; Ling, Xuefeng B.

    2016-01-01

    Objectives Kawasaki disease (KD) is an acute pediatric vasculitis of infants and young children with unknown etiology and no specific laboratory-based test to identify. A specific molecular diagnostic test is urgently needed to support the clinical decision of proper medical intervention, preventing subsequent complications of coronary artery aneurysms. We used a simple and low-cost colorimetric sensor array to address the lack of a specific diagnostic test to differentiate KD from febrile control (FC) patients with similar rash/fever illnesses. Study Design Demographic and clinical data were prospectively collected for subjects with KD and FCs under standard protocol. After screening using a genetic algorithm, eleven compounds including metalloporphyrins, pH indicators, redox indicators and solvatochromic dye categories, were selected from our chromatic compound library (n = 190) to construct a colorimetric sensor array for diagnosing KD. Quantitative color difference analysis led to a decision-tree-based KD diagnostic algorithm. Results This KD sensing array allowed the identification of 94% of KD subjects (receiver operating characteristic [ROC] area under the curve [AUC] 0.981) in the training set (33 KD, 33 FC) and 94% of KD subjects (ROC AUC: 0.873) in the testing set (16 KD, 17 FC). Color difference maps reconstructed from the digital images of the sensing compounds demonstrated distinctive patterns differentiating KD from FC patients. Conclusions The colorimetric sensor array, composed of common used chemical compounds, is an easily accessible, low-cost method to realize the discrimination of subjects with KD from other febrile illness. PMID:26859297

  8. Exhaled Breath Analysis with a Colorimetric Sensor Array for the Identification and Characterization of Lung Cancer

    PubMed Central

    Mazzone, Peter J; Wang, Xiao-Feng; Xu, Yaomin; Mekhail, Tarek; Beukemann, Mary C; Na, Jie; Kemling, Jonathan W; Suslick, Kenneth S; Sasidhar, Madhu

    2011-01-01

    Introduction The pattern of exhaled breath volatile organic compounds represents a metabolic biosignature with the potential to identify and characterize lung cancer. Breath biosignature-based classification of homogeneous subgroups of lung cancer may be more accurate than a global breath signature. Combining breath biosignatures with clinical risk factors may improve the accuracy of the signature. Objectives Develop an exhaled breath biosignature of lung cancer using a colorimetric sensor array. Determine the accuracy of breath biosignatures of lung cancer characteristics with and without the inclusion of clinical risk factors. Methods The exhaled breath of 229 study subjects, 92 with lung cancer and 137 controls, was drawn across a colorimetric sensor array. Logistic prediction models were developed and statistically validated based on the color changes of the sensor. Age, sex, smoking history, and COPD were incorporated in the prediction models. Results The validated prediction model of the combined breath and clinical biosignature was moderately accurate at distinguishing lung cancer from control subjects (C-statistic 0.811). The accuracy improved when the model focused on only one histology (C-statistic 0.825 – 0.890). Individuals with different histologies could be accurately distinguished from one another (C-statistic 0.889 for adenocarcinoma vs. squamous cell carcinoma). Moderate accuracies were noted for validated breath biosignatures of stage and survival (C-statistic 0.793, 0.770 respectively). Conclusions A colorimetric sensor array is capable of identifying exhaled breath biosignatures of lung cancer. The accuracy of breath biosignatures can be optimized by evaluating specific histologies and incorporating clinical risk factors. PMID:22071780

  9. Evaluation of micro-colorimetric lipid determination method with samples prepared using sonication and accelerated solvent extraction methods.

    PubMed

    Billa, Nanditha; Hubin-Barrows, Dylan; Lahren, Tylor; Burkhard, Lawrence P

    2014-02-01

    Two common laboratory extraction techniques were evaluated for routine use with the micro-colorimetric lipid determination method developed by Van Handel (1985) [2] and recently validated for small samples by Inouye and Lotufo (2006) [1]. With the accelerated solvent extraction method using chloroform:methanol solvent and the colorimetric lipid determination method, 28 of 30 samples had significant proportional bias (α=1%, determined using standard additions) and 1 of 30 samples had significant constant bias (α=1%, determined using Youden Blank measurements). With sonic extraction, 0 of 6 samples had significant proportional bias (α=1%) and 1 of 6 samples had significant constant bias (α=1%). These demonstrate that the accelerated solvent extraction method with chloroform:methanol solvent system creates an interference with the colorimetric assay method, and without accounting for the bias in the analysis, inaccurate measurements would be obtained. PMID:24401464

  10. Data set of optimal parameters for colorimetric red assay of epoxide hydrolase activity.

    PubMed

    de Oliveira, Gabriel Stephani; Adriani, Patricia Pereira; Borges, Flavia Garcia; Lopes, Adriana Rios; Campana, Patricia T; Chambergo, Felipe S

    2016-09-01

    The data presented in this article are related to the research article entitled "Epoxide hydrolase of Trichoderma reesei: Biochemical properties and conformational characterization" [1]. Epoxide hydrolases (EHs) are enzymes that catalyze the hydrolysis of epoxides to the corresponding vicinal diols. This article describes the optimal parameters for the colorimetric red assay to determine the enzymatic activity, with an emphasis on the characterization of the kinetic parameters, pH optimum and thermal stability of this enzyme. The effects of reagents that are not resistant to oxidation by sodium periodate on the reactions can generate false positives and interfere with the final results of the red assay. PMID:27366781

  11. Colorimetric determination of ozone in water based on reaction with bis(terpyridine)iron(II)

    SciTech Connect

    Tomiyasu, H.; Gordon, G.

    1984-04-01

    The analysis of aqueous ozone was studied by a colorimetric method that uses the reaction between ozone and bis(terpyridine)iron(II), Fe(terpy)/sub 2//sup 2 +/, in dilute hydrochloric acid solution. The ozone concentrations were determined from the change in absorption spectra of Fe(terpy)/sub 2//sup 2 +/ at concentrations as low as 10/sup -6/ M (0.05 mg/L) with an accuracy and precision of better than +/-3%. Direct comparisons with the Indigo method and UV spectrophotometric measurements are presented.

  12. Highly Sensitive and Fast Response Colorimetric Humidity Sensors Based on Graphene Oxides Film.

    PubMed

    Chi, Hong; Liu, Yan Jun; Wang, FuKe; He, Chaobin

    2015-09-16

    Uniform graphene oxide (GO) film for optical humidity sensing was fabricated by dip-coating technique. The resulting GO thin film shows linear optical shifts in the visible range with increase of humidity in the whole relative humidity range (from dry state to 98%). Moreover, GO films exhibit ultrafast sensing to moisture within 250 ms because of the unique atomic thinness and superpermeability of GO sheets. The humidity sensing mechanism was investigated using XRD and computer simulation. The ultrasensitive humidity colorimetric properties of GOs film may enable many potential applications such as disposable humidity sensors for packaging, health, and environmental monitoring. PMID:26305842

  13. Colorimetric response of azobenzene-terminated polydiacetylene vesicles under thermal and photic stimuli

    NASA Astrophysics Data System (ADS)

    You, Xian; Chen, Xin; Zou, Gang; Su, Wei; Zhang, Qijin; He, Pingsheng

    2009-11-01

    We study the colorimetric reversibility of pure polymerized p-nitro azobenzene moiety-substituted diacetylene (PNADA) and PNADA/polymerized 10,12-pentacosadiynoic acid (PDA) complex vesicles under thermal and photic stimuli. Because of the strong intermolecular interaction among azobenzene mesogens within the vesicles, PNADA vesicles show enhanced stability and completely reversible thermochromic response. Polydiacetylene based complex vesicles with partial reversible chromatic properties under both thermal and photonic stimuli were reported for the first time, which provided a novel model system for the understanding of the chromatic transition mechanism of polydiacetylene materials.

  14. A colorimetric and fluorescent dual probe for palladium in aqueous medium and live cell imaging.

    PubMed

    Yan, Jin-Wu; Wang, Xiao-Lin; Tan, Qi-Feng; Yao, Pei-Fen; Tan, Jia-Heng; Zhang, Lei

    2016-04-21

    A colorimetric and fluorescent dual probe for palladium species was rationally developed by combining the resorufin fluorophore with allyl chloroformate. The probe enables the visual detection of palladium based on its vivid color change from pale yellow to pink and its fluorescence off-on response to palladium in PBS solution. The detection limit was calculated to be as low as 2.1 nM. The live cell imaging results showed that this probe could be used as an effective fluorescent probe for detecting intracellular palladium species. All these results featured its promising application prospects in the palladium analytical field. PMID:26990285

  15. Spectroscopic and computational study of a naphthalene derivative as colorimetric and fluorescent sensor for bioactive anions.

    PubMed

    Sharma, Darshna; Sahoo, Suban K; Bera, Rati Kanta; Kamal, Raj

    2013-05-01

    The anion recognition property of a naphthalene based receptor (L) was investigated by naked-eye, UV-Vis, fluorescence, (1)H NMR and computational methods. The receptor L showed fluoride selective naked-eye detectable colorimetric and UV-Vis spectral changes over other tested anions due to the formation of hydrogen bonding complex in 1:1 stoichiometry and/or deprotonation between fluoride and the receptor. Interestingly, the fluorescence of L was quenched by fluoride but enhanced by acetate. PMID:23456416

  16. Colorimetric and microscopic study of the thermal behavior of new ceramic pigments.

    PubMed

    Saviuc-Paval, Ana Mihaela; Victor Sandu, Andrei; Marcel Popa, Ionel; Anca Sandu, Irina Crina; Petru Bertea, Andrei; Sandu, Ion

    2013-06-01

    The article studies thermal resistance variation by analyzing the colorimetric parameters correlated with the optical microscopy data of two groups of ceramic pigments obtained by co-precipitation in aqueous medium of phosphate anion and of a mixture of chromium phosphate with various chromophore cations. This research enabled us to reveal the thermal thresholds/domains within which significant color changes occur, thus allowing the choice of pigments compatible with the thermal varnishing-glazing processes involved in the manufacture of tesserae for mosaic and stained glass and of colored materials for floor tiles, wall tiles and painted porcelain. PMID:23495175

  17. Dicyclohexylcarbodiimide as a cleaving agent for colorimetric determination of pyridyl and pyrimidinyl compounds

    SciTech Connect

    Chen, S.C.

    1985-06-01

    By use of dicyclohexylcarbodiimide (DCC) and dimethylbarbituric acid (DMBA) as reagents, a colorimetric method for the determination of pyridyl and pyrimidinyl compounds has been established. DCC breaks the pyridine or pyrimidine ring to afford glutaconaldehyde or malonaldehyde and then reacts with DMBA to produce chromophores. These heterocycles could be determined by measuring the chromophores. The relative standard deviations obtained with different amounts of these compounds were in a range of 0.63 to 5.36% (n = 10). The reaction mechanism is also discussed. 22 references, 4 figures, 1 table.

  18. A novel colorimetric and fluorescent sensor based on calix[4]arene possessing triphenylamine units

    NASA Astrophysics Data System (ADS)

    Erdemir, Serkan; Malkondu, Sait; Kocyigit, Ozcan; Alıcı, Onder

    2013-10-01

    A novel colorimetric and fluorometric calix[4]arene probe (CTP) bearing triphenylamine units was synthesized in good yield and characterized by combination of 1H, 13C, APT, COSY, FTIR, HRMS, and UV-vis spectral data. Ion-binding studies of CTP were investigated in acetonitrile with a wide range of cations and anions and the recognition process was monitored by luminescence, UV-vis and 1H NMR spectral changes. CTP exhibited naked eye detection for Hg2+ ion. Also it showed a significant fluorescence quenching towards F- ion.

  19. Gold-Nanoparticle-Based Colorimetric Sensor Array for Discrimination of Organophosphate Pesticides.

    PubMed

    Fahimi-Kashani, Nafiseh; Hormozi-Nezhad, M Reza

    2016-08-16

    There is a growing interest in developing high-performance sensors monitoring organophosphate pesticides, primarily due to their broad usage and harmful effects on mammals. In the present study, a colorimetric sensor array consisting of citrate-capped 13 nm gold nanoparticles (AuNPs) has been proposed for the detection and discrimination of several organophosphate pesticides (OPs). The aggregation-induced spectral changes of AuNPs upon OP addition has been analyzed with pattern recognition techniques, including hierarchical cluster analysis (HCA) and linear discriminant analysis (LDA). In addition, the proposed sensor array has the capability to identify individual OPs or mixtures of them in real samples. PMID:27412472

  20. An instantaneous colorimetric protein assay based on spontaneous formation of a protein corona on gold nanoparticles.

    PubMed

    Ho, Yan Teck; Poinard, Barbara; Yeo, Eugenia Li Ling; Kah, James Chen Yong

    2015-02-21

    Commercial protein assays used ubiquitously in laboratories typically require long incubation times due to the inherently slow protein-reagent reactions. In this study, we report a novel facile technique for the instantaneous measurement of total protein concentration by exploiting the rapid aggregation dynamics of gold nanoparticles (NPs). By adsorbing different amounts of proteins on their surface to form a protein corona, these NPs can be sterically stabilized to different degrees by aggregation, thus exhibiting a spectrum of color change which can be quantitatively characterized by UV-Vis absorption spectroscopy. We evaluated this technique on four model proteins with different structures: bovine serum albumin (BSA), normal mouse immunoglobulin G (IgG), fibrinogen (FBG) and apolipoprotein A-I (Apo-A1) using two approaches, sequential and simultaneous. We obtained an approach-dependent linear concentration range up to 80 μg mL(-1) and 400 μg mL(-1) for sequential and simultaneous approaches, respectively. This linear working range was wider than that of the commercial Bradford assay and comparable to the Micro BCA assay. The simultaneous approach was also able to produce a linear working range of 200 to 1000 μg mL(-1) (R(2) = 0.995) in human urine, while the sequential approach was non-functional in urine. Similar to Micro BCA, the NP-based protein assay was able to elicit a linear response (R(2) > 0.87) for all four proteins with different structures. However, unlike Micro BCA which requires up to 120 min of incubation, we were able to obtain the read-out almost instantaneously without the need for incubation. The NP-based technique using the simultaneous approach can thus be exploited as a novel assay for instantaneous protein quantification to increase the productivity of laboratory processes. PMID:25501998