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Sample records for rapid colorimetric quantification

  1. 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

  2. Colorimetric method for rapid determination of bacteriuria.

    PubMed

    Wallis, C; Melnick, J L; Longoria, C J

    1981-09-01

    An inexpensive, rapid, and simple colorimetric test for detection of bacteriuria is described. This test does not require bacterial growth and has the marked advantage of being able to quantify bacteria, even when the organisms are present in the urine of bacteriuric patients who are being treated with antibiotics. The test is carried out with 1 ml of urine, which is processed through a 10-mm-diameter filter than entraps the bacteria on its surface. Safranine dye is passed through the filter to stain the bacteria and the filter fibers. A decolorizer, which removes the dye from the filter fibers but not from the bacteria, is then passed through the filter. If there are greater than or equal to 10(5) colony-forming units of bacteria per ml in the sample, the 10-mm filter disk manifests a pink to red color. If there are less than 10(5)colony-forming units of bacteria per ml, the filter disk remains white or becomes slightly yellow. The entire procedure has been adapted to a semiautomated instrument and the time required per test is less than 1 min. The results obtained on the test card are a permanent record to be filled with the patient's chart. The bacteria can be quickly classified as gram positive or gram negative by selective staining of a second milliliter of urine on the filter. Of 441 urine specimens tested, 430 (98%) were correctly classified as containing more or less than 10(5) colony-forming units per ml. A total of 62 urine specimens were positive by bacterial plating (greater than or equal to 10(5) colony-forming units per ml), and 59 were positive by the colorimetric test. Eight false-positives (colorimetric test positive, plate counts less than 10(5)) were encountered in patients (bacteriuric) being prescribed antibiotics. Removal of the antibiotics from these urine specimens, with subsequent replating of the samples, indicated the presence of greater than or equal to 10(5) colony-forming units of bacteria per ml in three representative cases tested

  3. Critical coagulation concentration-based salt titration for visual quantification in gold nanoparticle-based colorimetric biosensors.

    PubMed

    Li, Na; Yu, Lu; Zou, Jiaqi

    2014-02-01

    In gold nanoparticle (GNP)-based colorimetric biosensors, the gradual color shift is often used to correlate with the concentration of target molecules, and therefore a UV-vis spectrometer is usually required for accurate quantification. Here, we present a critical coagulation concentration (CCC)-based salt titration as a generic and simple way to enable accurate quantification in GNP-based colorimetric biosensors without any analytical equipment. The titration is carried out by stepwise addition of a salt titrant to the premixture of sample and GNPs until the color changes rapidly from red to blue, determined solely by visual inspection. The number of titration steps or the final salt concentration required for a rapid color shift (i.e., CCC) is then used to quantitatively correlate with the concentration of target molecules in the sample. The salt titration-based quantification has been demonstrated with two previously reported GNP-based colorimetric biosensors. Compared with quantification based on the gradual color shift with a spectrometer, the visual quantification based on the rapid color shift in the salt titration eliminates the need for any analytical equipment without sacrificing the performance (i.e., sensitivity and accuracy) and therefore is highly suitable for applications in low-resource settings.

  4. 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

  5. Rapid and portable electrochemical quantification of phosphorus.

    PubMed

    Kolliopoulos, Athanasios V; Kampouris, Dimitrios K; Banks, Craig E

    2015-04-21

    Phosphorus is one of the key indicators of eutrophication levels in natural waters where it exists mainly as dissolved phosphorus. Various analytical protocols exist to provide an offsite analysis, and a point of site analysis is required. The current standard method recommended by the Environmental Protection Agency (EPA) for the detection of total phosphorus is colorimetric and based upon the color of a phosphomolybdate complex formed as a result of the reaction between orthophosphates and molybdates ions where ascorbic acid and antimony potassium tartrate are added and serve as reducing agents. Prior to the measurements, all forms of phosphorus are converted into orthophosphates via sample digestion (heating and acidifying). The work presented here details an electrochemical adaptation of this EPA recommended colorimetric approach for the measurement of dissolved phosphorus in water samples using screen-printed graphite macroelectrodes for the first time. This novel indirect electrochemical sensing protocol allows the determination of orthophosphates over the range from 0.5 to 20 μg L(-1) in ideal pH 1 solutions utilizing cyclic voltammetry with a limit of detection (3σ) found to correspond to 0.3 μg L(-1) of phosphorus. The reaction time and influence of foreign ions (potential interferents) upon this electroanalytical protocol was also investigated, where it was found that a reaction time of 5 min, which is essential in the standard colorimetric approach, is not required in the new proposed electrochemically adapted protocol. The proposed electrochemical method was independently validated through the quantification of orthophosphates and total dissolved phosphorus in polluted water samples (canal water samples) with ion chromatography and ICP-OES, respectively. This novel electrochemical protocol exhibits advantages over the established EPA recommended colorimetric determination for total phosphorus with lower detection limits and shorter experimental times

  6. Rapid and portable electrochemical quantification of phosphorus.

    PubMed

    Kolliopoulos, Athanasios V; Kampouris, Dimitrios K; Banks, Craig E

    2015-04-21

    Phosphorus is one of the key indicators of eutrophication levels in natural waters where it exists mainly as dissolved phosphorus. Various analytical protocols exist to provide an offsite analysis, and a point of site analysis is required. The current standard method recommended by the Environmental Protection Agency (EPA) for the detection of total phosphorus is colorimetric and based upon the color of a phosphomolybdate complex formed as a result of the reaction between orthophosphates and molybdates ions where ascorbic acid and antimony potassium tartrate are added and serve as reducing agents. Prior to the measurements, all forms of phosphorus are converted into orthophosphates via sample digestion (heating and acidifying). The work presented here details an electrochemical adaptation of this EPA recommended colorimetric approach for the measurement of dissolved phosphorus in water samples using screen-printed graphite macroelectrodes for the first time. This novel indirect electrochemical sensing protocol allows the determination of orthophosphates over the range from 0.5 to 20 μg L(-1) in ideal pH 1 solutions utilizing cyclic voltammetry with a limit of detection (3σ) found to correspond to 0.3 μg L(-1) of phosphorus. The reaction time and influence of foreign ions (potential interferents) upon this electroanalytical protocol was also investigated, where it was found that a reaction time of 5 min, which is essential in the standard colorimetric approach, is not required in the new proposed electrochemically adapted protocol. The proposed electrochemical method was independently validated through the quantification of orthophosphates and total dissolved phosphorus in polluted water samples (canal water samples) with ion chromatography and ICP-OES, respectively. This novel electrochemical protocol exhibits advantages over the established EPA recommended colorimetric determination for total phosphorus with lower detection limits and shorter experimental times

  7. Quantification of detergent using colorimetric methods in membrane protein crystallography.

    PubMed

    Prince, Chelsy; Jia, Zongchao

    2015-01-01

    Membrane protein crystallography has the potential to greatly aid our understanding of membrane protein biology. Yet, membrane protein crystals remain challenging to produce. Although robust methods for the expression and purification of membrane proteins continue to be developed, the detergent component of membrane protein samples is equally important to crystallization efforts. This chapter describes the development of three colorimetric assays for the quantitation of detergent in membrane protein samples and provides detailed protocols. All of these techniques use small sample volumes and have potential applications in crystallography. The application of these techniques in crystallization prescreening, detergent concentration modification, and detergent exchange experiments is demonstrated. It has been observed that the concentration of detergent in a membrane protein sample can be just as important as the protein concentration when attempting to reproduce crystallization lead conditions.

  8. 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…

  9. 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...

  10. A simplified diphenylamine colorimetric method for growth quantification.

    PubMed

    Zhao, Youbao; Xiang, Sihai; Dai, Xida; Yang, Keqian

    2013-06-01

    Cell growth needs to be monitored in biological studies and bioprocess optimization. In special circumstances, such as microbial fermentations in media containing insoluble particles, accurate cell growth quantification is a challenge with current methods. Only the Burton method is applicable in such circumstances. The original Burton method was previously simplified by adopting a two-step sample pretreatment in perchloric acid procedure to eliminate the need for DNA extraction. Here, we further simplified the Burton method by replacing the previous two-step perchloric acid pretreatment with a new and one-step diphenylamine reagent pretreatment. The reliability and accuracy of this simplified method were assessed by measuring the biomass of four model microorganisms: Escherichia coli, Streptomyces clavuligerus, Saccharomyces cerevisiae, and Trichoderma reesei grown in normal media or those containing solid particles. The results demonstrate that this new simplified method performs comparably to the conventional methods, such as OD600 or the previously modified Burton method, and is much more sensitive than the dry weight method. Overall, the new method is simple, reliable, easy to perform, and generally applicable in most circumstances, and it reduces the operation time from more than 12 h (for the previously simplified Burton method) to about 2 h.

  11. Comparison of colorimetric assays with quantitative amino acid analysis for protein quantification of Generalized Modules for Membrane Antigens (GMMA).

    PubMed

    Rossi, Omar; Maggiore, Luana; Necchi, Francesca; Koeberling, Oliver; MacLennan, Calman A; Saul, Allan; Gerke, Christiane

    2015-01-01

    Genetically induced outer membrane particles from Gram-negative bacteria, called Generalized Modules for Membrane Antigens (GMMA), are being investigated as vaccines. Rapid methods are required for estimating the protein content for in-process assays during production. Since GMMA are complex biological structures containing lipid and polysaccharide as well as protein, protein determinations are not necessarily straightforward. We compared protein quantification by Bradford, Lowry, and Non-Interfering assays using bovine serum albumin (BSA) as standard with quantitative amino acid (AA) analysis, the most accurate currently available method for protein quantification. The Lowry assay has the lowest inter- and intra-assay variation and gives the best linearity between protein amount and absorbance. In all three assays, the color yield (optical density per mass of protein) of GMMA was markedly different from that of BSA with a ratio of approximately 4 for the Bradford assay, and highly variable between different GMMA; and approximately 0.7 for the Lowry and Non-Interfering assays, highlighting the need for calibrating the standard used in the colorimetric assay against GMMA quantified by AA analysis. In terms of a combination of ease, reproducibility, and proportionality of protein measurement, and comparability between samples, the Lowry assay was superior to Bradford and Non-Interfering assays for GMMA quantification.

  12. 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.

  13. Colorimetric Focus-Forming Assay with Automated Focus Counting by Image Analysis for Quantification of Infectious Hepatitis C Virions

    PubMed Central

    Kang, Wonseok; Shin, Eui-Cheol

    2012-01-01

    Hepatitis C virus (HCV) infection is the leading cause of liver transplantation in Western countries. Studies of HCV infection using cell culture-produced HCV (HCVcc) in vitro systems require quantification of infectious HCV virions, which has conventionally been performed by immunofluorescence-based focus-forming assay with manual foci counting; however, this is a laborious and time-consuming procedure with potentially biased results. In the present study, we established and optimized a method for convenient and objective quantification of HCV virions by colorimetric focus-forming assay with automated focus counting by image analysis. In testing different enzymes and chromogenic substrates, we obtained superior foci development using alkaline phosphatase-conjugated secondary antibody with BCIP/NBT chromogenic substrate. We additionally found that type I collagen coating minimized cell detachment during vigorous washing of the assay plate. After the colorimetric focus-forming assay, the foci number was determined using an ELISpot reader and image analysis software. The foci number and the calculated viral titer determined by this method strongly correlated with those determined by immunofluorescence-based focus-forming assay and manual foci counting. These results indicate that colorimetric focus-forming assay with automated focus counting by image analysis is applicable as a more-efficient and objective method for quantification of infectious HCV virions. PMID:22937136

  14. 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

  15. A gold nanoparticle-based colorimetric probe for rapid detection of 1-hydroxypyrene in urine.

    PubMed

    Hu, Yaqi; Du, Chunyan; Li, Yunchao; Fan, Louzhen; Li, Xiaohong

    2015-07-01

    Direct and rapid detection of 1-hydroxypyrene (1-OHP) is of great importance owing to its high carcinogenicity, teratogenicity and toxicity. In this study, a simple colorimetric assay for rapid determination of 1-OHP is reported, which is based on non-crosslinking aggregation of gold nanoparticles (Au NPs) induced by 1-OHP in the presence of formic acid (FA). Initially, Au NPs were synthesized with citrate as the capping agent and exhibited red color. Subsequently, the addition of FA did not cause aggregation of Au NPs, but a proton transfer process occurred from FA to carboxylic anions on the surface of Au NPs with a decreased zeta potential. The subsequent addition of 1-OHP resulted in a further decreased zeta potential and an intensely hydrophobic environment, which led to a strong and rapid non-crosslinking aggregation of Au NPs within 5 min with the color changing from red to violet blue. Based on this principle, sensitive and selective detection of 1-OHP was achieved. The detection limit was 3.3 nM. Finally, the colorimetric assay was successfully applied to detect 1-OHP in a urine sample. This strategy provides new insights into developing colorimetric methods for on-site and real-time detection of polycyclic aromatic hydrocarbons. PMID:25988203

  16. 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

  17. 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

  18. 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

    2016-05-02

    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.

  19. 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

  20. 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

  1. A rapid, simple method for determining formaldehyde in drinking water using colorimetric-solid phase extraction.

    PubMed

    Hill, April A; Lipert, Robert J; Fritz, James S; Porter, Marc D

    2009-02-15

    Formaldehyde has been detected in drinking water supplies across the globe and on board NASA spacecraft. A rapid, simple, microgravity-compatible technique for measuring this contaminant in water supplies using colorimetric-solid phase extraction (C-SPE) is described. This method involves collecting a water sample into a syringe by passage through a cartridge that contains sodium hydroxide, to adjust pH, and Purpald, which is a well-established colorimetric reagent for aldehydes. After completing the reaction in the syringe by agitating for 2 min on a shaker at 400 rpm, the 1.0-mL alkaline sample is passed through an extraction disk that retains the purple product. The amount of concentrated product is then measured on-disk using diffuse reflectance spectroscopy, and compared to a calibration plot generated from Kubelka-Munk transformations of the reflectance data at 700 nm to determine the formaldehyde concentration. This method is capable of determining formaldehyde concentrations from 0.08 to 20 ppm with a total work-up time of less than 3 min using only 1-mL samples.

  2. 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

  3. Molecularly imprinted photonic hydrogels as colorimetric sensors for rapid and label-free detection of vanillin.

    PubMed

    Peng, Hailong; Wang, Shenqi; Zhang, Zhong; Xiong, Hua; Li, Jinhua; Chen, Lingxin; Li, Yanbin

    2012-02-29

    A novel colorimetric sensor for the rapid and label-free detection of vanillin, based on the combination of photonic crystal and molecular imprinting technique, was developed. The sensing platform of molecularly imprinted photonic hydrogel (MIPH) was prepared by a noncovalent and self-assembly approach using vanillin as a template molecule. Morphology characterization by scanning electron microscope (SEM) showed that the MIPH possessed a highly ordered three-dimensional (3D) macroporous structure with nanocavities. The vanillin recognition events of the created nonocavities could be directly transferred into readable optical signals through a change in Bragg diffraction of the ordered macropores array of MIPH. The Bragg diffraction peak shifted from 451 to 486 nm when the concentration of the vanillin was increased from 10⁻¹² to 10⁻³ mol L⁻¹ within 60 s, whereas there were no obvious peak shifts for methyl and ethyl vanillin, indicating that the MIPH had high selectivity and rapid response for vanillin. The adsorption results showed that the hierarchical porous structure and homogeneous layers were formed in the MIPH with higher adsorption capacity. The application of such a label-free sensor with high selectivity, high sensitivity, high stability, and easy operation might offer a potential method for rapid real-time detection of trace vanillin.

  4. 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.

  5. 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.

  6. 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

  7. Rapid colorimetric sensing platform for the detection of Listeria monocytogenes foodborne pathogen.

    PubMed

    Alhogail, Sahar; Suaifan, Ghadeer A R Y; Zourob, Mohammed

    2016-12-15

    Listeria monocytogenes is a serious cause of human foodborne infections worldwide, which needs spending billions of dollars for inspection of bacterial contamination in food every year. Therefore, there is an urgent need for rapid, in-field and cost effective detection techniques. In this study, rapid, low-cost and simple colorimetric assay was developed using magnetic nanoparticles for the detection of listeria bacteria. The protease from the listeria bacteria was detected using D-amino acid substrate. D-amino acid substrate was linked to the carboxylic acid on the magnetic nanoparticles using EDC/NHS chemistry. The cysteine residue at the C-terminal of the substrate was used for the self-assembled monolayer formation on the gold sensor surface, which in turn the black magnetic nanobeads will mask the golden color. The color will change from black to golden color upon the cleavage of the specific peptide sequence by the Listeria protease. The sensor was tested with serial dilutions of Listeria bacteria. It was found that the appearance of the gold surface area is proportional to the bacterial concentrations in CFU/ml. The lowest detection limit of the developed sensor for Listeria was found to be 2.17×10(2) colony forming unit/ml (CFU/ml). The specificity of the biosensor was tested against four different foodborne associated bacteria (Escherichia coli, Salmonella, Shigella flexnerii and Staphylococcus aureus). Finally, the sensor was tested with artificially spiked whole milk and ground meat spiked with listeria. PMID:27543841

  8. Rapid colorimetric sensing platform for the detection of Listeria monocytogenes foodborne pathogen.

    PubMed

    Alhogail, Sahar; Suaifan, Ghadeer A R Y; Zourob, Mohammed

    2016-12-15

    Listeria monocytogenes is a serious cause of human foodborne infections worldwide, which needs spending billions of dollars for inspection of bacterial contamination in food every year. Therefore, there is an urgent need for rapid, in-field and cost effective detection techniques. In this study, rapid, low-cost and simple colorimetric assay was developed using magnetic nanoparticles for the detection of listeria bacteria. The protease from the listeria bacteria was detected using D-amino acid substrate. D-amino acid substrate was linked to the carboxylic acid on the magnetic nanoparticles using EDC/NHS chemistry. The cysteine residue at the C-terminal of the substrate was used for the self-assembled monolayer formation on the gold sensor surface, which in turn the black magnetic nanobeads will mask the golden color. The color will change from black to golden color upon the cleavage of the specific peptide sequence by the Listeria protease. The sensor was tested with serial dilutions of Listeria bacteria. It was found that the appearance of the gold surface area is proportional to the bacterial concentrations in CFU/ml. The lowest detection limit of the developed sensor for Listeria was found to be 2.17×10(2) colony forming unit/ml (CFU/ml). The specificity of the biosensor was tested against four different foodborne associated bacteria (Escherichia coli, Salmonella, Shigella flexnerii and Staphylococcus aureus). Finally, the sensor was tested with artificially spiked whole milk and ground meat spiked with listeria.

  9. Rapid and Automated Quantification of Microalgal Lipids on a Spinning Disc.

    PubMed

    Kim, Yubin; Jeong, Su-Nam; Kim, Bolam; Kim, Dong-Pyo; Cho, Yoon-Kyoung

    2015-08-01

    We have developed a fully integrated centrifugal microfluidic device for rapid on-site quantification of lipids from microalgal samples. The fully automated serial process involving cell sedimentation and lysis, liquid-liquid extraction, and colorimetric detection of lipid contents was accomplished within 13 min using a lab-on-a-disc. The presented organic solvent-tolerable (for n-hexane, ethanol) microfluidic disc was newly fabricated by combining thermal fusion bonding and carbon dot-based valving techniques. It is expected that this novel platform will possibly contribute toward sustainable biofuel applications by providing a practical solution for on-site monitoring of lipid accumulation in microalgal samples, thus providing imperative contribution toward energy and environmental purposes of centrifugal microfluidic technology.

  10. Rapid Colorimetric Assays to Qualitatively Distinguish RNA and DNA in Biomolecular Samples

    PubMed Central

    Patterson, Jennifer; Mura, Cameron

    2013-01-01

    Biochemical experimentation generally requires accurate knowledge, at an early stage, of the nucleic acid, protein, and other biomolecular components in potentially heterogeneous specimens. Nucleic acids can be detected via several established approaches, including analytical methods that are spectrophotometric (e.g., A260), fluorometric (e.g., binding of fluorescent dyes), or colorimetric (nucleoside-specific chromogenic chemical reactions).1 Though it cannot readily distinguish RNA from DNA, the A260/A280 ratio is commonly employed, as it offers a simple and rapid2 assessment of the relative content of nucleic acid, which absorbs predominantly near 260 nm and protein, which absorbs primarily near 280 nm. Ratios < 0.8 are taken as indicative of 'pure' protein specimens, while pure nucleic acid (NA) is characterized by ratios > 1.53. However, there are scenarios in which the protein/NA content cannot be as clearly or reliably inferred from simple uv-vis spectrophotometric measurements. For instance, (i) samples may contain one or more proteins which are relatively devoid of the aromatic amino acids responsible for absorption at ≈280 nm (Trp, Tyr, Phe), as is the case with some small RNA-binding proteins, and (ii) samples can exhibit intermediate A260/A280 ratios (~0.8 < ~1.5), where the protein/NA content is far less clear and may even reflect some high-affinity association between the protein and NA components. For such scenarios, we describe herein a suite of colorimetric assays to rapidly distinguish RNA, DNA, and reducing sugars in a potentially mixed sample of biomolecules. The methods rely on the differential sensitivity of pentoses and other carbohydrates to Benedict's, Bial's (orcinol), and Dische's (diphenylamine) reagents; the streamlined protocols can be completed in a matter of minutes, without any additional steps of having to isolate the components. The assays can be performed in parallel to differentiate between RNA and DNA, as well as indicate the

  11. 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

  12. 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.

  13. A novel colorimetric assay for rapid detection of cysteine and Hg²⁺ based on gold clusters.

    PubMed

    Wang, Yi-Wei; Tang, Shurong; Yang, Huang-Hao; Song, Hongbo

    2016-01-01

    Inhibition and recovery of the catalytic activity of bovine serum albumin-capped gold nanoclusters (BSA-AuNCs) is observed for the first time by introduction of cysteine and Hg(2+). The prepared BSA-AuNCs possess highly intrinsic peroxidase-like activity. It can catalyze the oxidation of 3, 3, 5, 5-tetramethylbenzidine by H2O2 to produce a blue colored product. Based on this phenomenon, a new colorimetric assay for rapid, selective and sensitive detection of cysteine and Hg(2+) in aqueous solution has been demonstrated. The interaction process between target molecule and BSA-AuNCs is very fast, so that the whole test can be completed within ten minutes. Moreover, the fabricated colorimetric sensor is simple and cost-effective, without the need of nucleic acid based recognition element and complicated washing, separation and labeling process, thus holds great promise for routine analysis of cysteine and Hg(2+) in real samples. PMID:26695236

  14. A novel colorimetric assay for rapid detection of cysteine and Hg²⁺ based on gold clusters.

    PubMed

    Wang, Yi-Wei; Tang, Shurong; Yang, Huang-Hao; Song, Hongbo

    2016-01-01

    Inhibition and recovery of the catalytic activity of bovine serum albumin-capped gold nanoclusters (BSA-AuNCs) is observed for the first time by introduction of cysteine and Hg(2+). The prepared BSA-AuNCs possess highly intrinsic peroxidase-like activity. It can catalyze the oxidation of 3, 3, 5, 5-tetramethylbenzidine by H2O2 to produce a blue colored product. Based on this phenomenon, a new colorimetric assay for rapid, selective and sensitive detection of cysteine and Hg(2+) in aqueous solution has been demonstrated. The interaction process between target molecule and BSA-AuNCs is very fast, so that the whole test can be completed within ten minutes. Moreover, the fabricated colorimetric sensor is simple and cost-effective, without the need of nucleic acid based recognition element and complicated washing, separation and labeling process, thus holds great promise for routine analysis of cysteine and Hg(2+) in real samples.

  15. 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.

  16. Rapid Quantification and Validation of Lipid Concentrations within Liposomes.

    PubMed

    Roces, Carla B; Kastner, Elisabeth; Stone, Peter; Lowry, Deborah; Perrie, Yvonne

    2016-01-01

    Quantification of the lipid content in liposomal adjuvants for subunit vaccine formulation is of extreme importance, since this concentration impacts both efficacy and stability. In this paper, we outline a high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) method that allows for the rapid and simultaneous quantification of lipid concentrations within liposomal systems prepared by three liposomal manufacturing techniques (lipid film hydration, high shear mixing, and microfluidics). The ELSD system was used to quantify four lipids: 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), cholesterol, dimethyldioctadecylammonium (DDA) bromide, and ᴅ-(+)-trehalose 6,6'-dibehenate (TDB). The developed method offers rapidity, high sensitivity, direct linearity, and a good consistency on the responses (R² > 0.993 for the four lipids tested). The corresponding limit of detection (LOD) and limit of quantification (LOQ) were 0.11 and 0.36 mg/mL (DMPC), 0.02 and 0.80 mg/mL (cholesterol), 0.06 and 0.20 mg/mL (DDA), and 0.05 and 0.16 mg/mL (TDB), respectively. HPLC-ELSD was shown to be a rapid and effective method for the quantification of lipids within liposome formulations without the need for lipid extraction processes. PMID:27649231

  17. Rapid Quantification and Validation of Lipid Concentrations within Liposomes

    PubMed Central

    Roces, Carla B.; Kastner, Elisabeth; Stone, Peter; Lowry, Deborah; Perrie, Yvonne

    2016-01-01

    Quantification of the lipid content in liposomal adjuvants for subunit vaccine formulation is of extreme importance, since this concentration impacts both efficacy and stability. In this paper, we outline a high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) method that allows for the rapid and simultaneous quantification of lipid concentrations within liposomal systems prepared by three liposomal manufacturing techniques (lipid film hydration, high shear mixing, and microfluidics). The ELSD system was used to quantify four lipids: 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), cholesterol, dimethyldioctadecylammonium (DDA) bromide, and d-(+)-trehalose 6,6′-dibehenate (TDB). The developed method offers rapidity, high sensitivity, direct linearity, and a good consistency on the responses (R2 > 0.993 for the four lipids tested). The corresponding limit of detection (LOD) and limit of quantification (LOQ) were 0.11 and 0.36 mg/mL (DMPC), 0.02 and 0.80 mg/mL (cholesterol), 0.06 and 0.20 mg/mL (DDA), and 0.05 and 0.16 mg/mL (TDB), respectively. HPLC-ELSD was shown to be a rapid and effective method for the quantification of lipids within liposome formulations without the need for lipid extraction processes. PMID:27649231

  18. Electrospun nanofiber based colorimetric probe for rapid detection of Fe2+ in water.

    PubMed

    Ondigo, D A; Tshentu, Z R; Torto, N

    2013-12-01

    An imidazole derivative, 2-(2'-pyridyl)imidazole (PIMH), was developed as a colorimetric probe for the qualitative analysis of Fe(2+) in aqueous solution. PIMH was then used to post-functionalize poly(vinylbenzyl chloride) (PVBC) nanofibers after electrospinning so as to afford a solid state colorimetric probe. Upon treatment with Fe(2+) the probe displayed a distinctive color change both in liquid and solid platforms. The linear dynamic range for the colorimetric determination of Fe(2+) was 0.0988-3.5 μg mL(-1). The ligand showed a high chromogenic selectivity for Fe(2+) over other cations with a detection limit of 0.102 μg mL(-1) in solution (lower than the WHO drinking water guideline limit of 2 mg L(-1)), and 2 μg mL(-1) in the solid state. The concentration of Fe(2+) in a certified reference material (Iron, Ferrous, 1072) was found to be 2.39±0.01 mg L(-1), which was comparable with the certified value of 2.44±0.12 mg L(-1). Application of the probe to real samples spiked with Fe(2+) achieved recoveries of over 97% confirming accuracy of the method and its potential for on-site monitoring. PMID:24267086

  19. Development, optimization and validation of a rapid colorimetric microplate bioassay for neomycin sulfate in pharmaceutical drug products.

    PubMed

    Francisco, Fabiane Lacerda; Saviano, Alessandro Morais; Pinto, Terezinha de Jesus Andreoli; Lourenço, Felipe Rebello

    2014-08-01

    Microbiological assays have been used to evaluate antimicrobial activity since the discovery of the first antibiotics. Despite their limitations, microbiological assays are widely employed to determine antibiotic potency of pharmaceutical dosage forms, since they provide a measure of biological activity. The aim of this work is to develop, optimize and validate a rapid colorimetric microplate bioassay for the potency of neomycin in pharmaceutical drug products. Factorial and response surface methodologies were used in the development and optimization of the choice of microorganism, culture medium composition, amount of inoculum, triphenyltetrazolium chloride (TTC) concentration and neomycin concentration. The optimized bioassay method was validated by the assessment of linearity (range 3.0 to 5.0μg/mL, r=0.998 and 0.994 for standard and sample curves, respectively), precision (relative standard deviation (RSD) of 2.8% and 4.0 for repeatability and intermediate precision, respectively), accuracy (mean recovery=100.2%) and robustness. Statistical analysis showed equivalency between agar diffusion microbiological assay and rapid colorimetric microplate bioassay. In addition, microplate bioassay had advantages concerning the sensitivity of response, time of incubation, and amount of culture medium and solutions required.

  20. A rapid and automated colorimetric assay for evaluating the sensitivity of herpes simplex strains to antiviral drugs.

    PubMed

    Langlois, M; Allard, J P; Nugier, F; Aymard, M

    1986-07-01

    A rapid and sensitive colorimetric assay has been developed to evaluate the sensitivity of herpes simplex viruses (HSV) to antiviral agents. The chessboard titration of viruses and antiviral drugs and the automatic reading and analysis of the data allows objective and accurate results to be rapidly obtained. Virus sensitivity was expressed as an ED50 value which was the concentration of drug (micrograms/ml) reducing viral cpe by 50%. The ED50 values of antiviral drugs [acetylguanosine (ACV), idoxuridine (IDU), deoxycytidine (IDC) and bromovinyl deoxyuridine] for several HSV reference strains were determined and the method was then applied to clinical specimens. The selection of ACV and IDU resistant mutants was performed on a cloned sensitive HSV 1 ocular strain. We observed cross-resistance between ACV and IDU for the mutants isolated. The resistance to thymidine-kinase-dependent antiviral agents varied in inverse ratio to the thymidine kinase activity induced by HSV strains.

  1. Rapid and selective lead (II) colorimetric sensor based on azacrown ether-functionalized gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Alizadeh, A.; Khodaei, M. M.; Karami, Ch; Workentin, M. S.; Shamsipur, M.; Sadeghi, M.

    2010-08-01

    A gold nanoparticle (AuNPs)-based simple and fast colorimetric sensor for selective detecting of Pb(II) in aqueous solution has been developed. Monodisperse AuNPs (approx. 2.0 nm diameter) has been prepared facilely and further modified with an alkanethiol-bearing monoazacrown ether terminus. These AuNPs are shown to selectively sense Pb2 + through color change, which is visually discernible by an appearance of the surface plasmon band (SPB) at 520 nm. The recognition mechanism is attributed to the unique structure of the monoazacrown ether attached to AuNPs and metal sandwich coordination between two azacrown ether moieties that are attached to separate nanoparticles. This inter-particle cross-linking results in an aggregation and apparent color change from brown to purple. Additionally, TEM experiments support the optical absorption data proving the aggregation between azacrown ether-capped gold nanoparticles. This AuNP-based colorimetric assay is a facile and robust method and allows fast detection of Pb2 + at ambient temperatures. More importantly, the developed technique does not utilize enzymatic reactions, light-sensitive dye molecules, lengthy protocols or sophisticated instrumentation.

  2. 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.

  3. 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

  4. Facile colorimetric method for simple and rapid detection of endotoxin based on counterion-mediated gold nanorods aggregation.

    PubMed

    Wang, Yashan; Zhang, Daohong; Liu, Wei; Zhang, Xiao; Yu, Shaoxuan; Liu, Tao; Zhang, Wentao; Zhu, Wenxin; Wang, Jianlong

    2014-05-15

    Existence of endotoxin in food and injection products indicates bacterial contaminations and therefore poses threat to human health. Herein, a simple and rapid colorimetric method for the effective detection of endotoxin in food and injections based on counterion-mediated gold nanorods aggregation is first proposed. By taking advantage of the color change of unmodified gold nanorods resulted from endotoxin mediated gold nanorods aggregation, endotoxin could be detected in the concentration range of 0.01-0.6 μM. Further, we studied the performance of gold nanorods with different aspect ratios (2.7 and 3.3) in determination of endotoxin and found that gold nanorods with higher aspect ratio (AR) showed superiority in the sensing sensitivity of endotoxin. A good specificity for endotoxin, a detection limit of 0.0084 μM and recoveries ranging from 84% to 109% in spiked food and injection samples are obtained with the colorimetric method. Results demonstrate that the present method provides a novel and effective approach for on-site screening of endotoxin in common products, which is beneficial for monitoring and reducing the risk of bacterial contaminations in food and injections production.

  5. On the photoacoustic, photothermal and colorimetric quantification of carotenoids and other phytonutrients in some foods: a review

    NASA Astrophysics Data System (ADS)

    Bicanic, Dane Danijel

    2011-05-01

    The performance of various analytical methods is compared in terms of their potentiality to quantify the concentration of carotenoids in some foods accurately and rapidly. High performance liquid chromatography (HPLC) and spectrophotometry (SP), two absolute reference techniques, were used in parallel experiments. The emphasis is on the application of the new methods to significantly reduce the long analysis time (due to a laborious and costly extraction) characteristic for both HPLC and spectrophotometry. Among such direct quantification methods are colorimetry and a variety of novel photoacoustic (PA) and photothermal (PT) detection schemes that obviate the extraction step. The outcome of numerous studies shows a number of important advantages provided by these methods. Furthermore, the promising results suggest that the availability of practical, versatile, compact and affordable quality control instruments that offer a low-cost solution to routine analysis of specific carotenoids in some foods is within reach.

  6. 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 ...

  7. 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

  8. 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.

  9. Development of a colorimetric assay for rapid quantitative measurement of clavulanic acid in microbial samples.

    PubMed

    Dai, Xida; Xiang, Sihai; Li, Jia; Gao, Qiang; Yang, Keqian

    2012-02-01

    We developed a colorimetric assay to quantify clavulanic acid (CA) in culture broth of Streptomyces clavuligerus, to facilitate screening of a large number of S. clavuligerus mutants. The assay is based on a β-lactamase-catalyzed reaction, in which the yellow substrate nitrocefin (λ (max)=390 nm) is converted to a red product (λ (max)=486 nm). Since CA can irreversibly inhibit β-lactamase activity, the level of CA in a sample can be measured as a function of the A (390)/A (486) ratio in the assay mixture. The sensitivity and detection window of the assay were determined to be 50 μg L(-1) and 50 μg L(-1) to 10 mg L(-1), respectively. The reliability of the assay was confirmed by comparing assay results with those obtained by HPLC. The assay was used to screen a pool of 65 S. clavuligerus mutants and was reliable for identifying CA over-producing mutants. Therefore, the assay saves time and labor in large-scale mutant screening and evaluation tasks. The detection window and the reliability of this assay are markedly better than those of previously reported CA assays. This assay method is suitable for high throughput screening of microbial samples and allows direct visual observation of CA levels on agar plates.

  10. 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

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

    2015-06-01

    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.

  11. 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

  12. 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

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

    2015-06-01

    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

  13. 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

  14. 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

  15. Rapid and simple colorimetric assay for detecting the enzymatic degradation of biodegradable plastic films.

    PubMed

    Shinozaki, Yukiko; Watanabe, Takashi; Nakajima-Kambe, Toshiaki; Kitamoto, Hiroko K

    2013-01-01

    We developed a rapid and simple method for evaluating the degradation of solid biodegradable plastics (BPs). Dye-containing BP films were used as substrates and the release of dye caused by the degradation of BPs was confirmed by a color change in the enzyme solution after a reaction time of 24 h.

  16. Evaluation of Colorimetric Methods Using Nicotinamide for Rapid Detection of Pyrazinamide Resistance in Mycobacterium tuberculosis▿

    PubMed Central

    Mirabal, Niuris C.; Yzquierdo, Sergio L.; Lemus, Dihadenys; Madruga, Mariela; Milián, Yoslaine; Echemendía, Miguel; Takiff, Howard; Martin, Anandi; Van der Stuyf, Patrick; Palomino, Juan Carlos; Montoro, Ernesto

    2010-01-01

    The direct detection of pyrazinamide resistance in Mycobacterium tuberculosis is sufficiently difficult that many laboratories do not attempt it. Most pyrazinamide resistance is caused by mutations that inactivate the pyrazinamidase enzyme needed to convert the prodrug pyrazinamide to its active form. We evaluated two newer and simpler methods to assess pyrazinamidase activity, the nitrate reductase and malachite green microtube assays, using nicotinamide in place of pyrazinamide. A total of 102 strains were tested by these methods and the results compared with those obtained by the classic Wayne assay. Mutations in the pncA gene were identified by sequencing the pncA genes from all isolates in which pyrazinamide resistance was detected by any of the three methods. Both the nitrate reductase and malachite green microtube assays showed sensitivities of 93.75% and specificities of 97.67%. Mutations in the pncA gene were found in 14 of 16 strains that were pyrazinamide resistant and in 1 of 4 strains that were sensitive by the Wayne assay. Both of these simple methods, used with nicotinamide, are promising and inexpensive alternatives for the rapid detection of pyrazinamide resistance in limited-resource countries. PMID:20554826

  17. 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...

  18. The determination of N-nitrosodiethanolamine (NDELA) at trace levels in shampoos and skin creams by a simple, rapid colorimetric method.

    PubMed

    Telling, G M; Dunnett, P C

    1981-10-01

    Synopsis A technique is described for the rapid determination of N-nitrosodiethanolamine (NDELA) in shampoos, skin creams and similar products based on aqueous extraction, partition into ethyl acetate and colorimetric determination using the Eisenbrand-Preussman cleavage reaction. Recoveries of NDELA added at levels of 5-100 mug kg(-1) to a range of shampoo and cream types ranged from 90-101%. The limit of determination for the method is 2.5 mug kg(-1). Observations on the application of a thermal energy analyser linked to a gas chromatograph are also reported. Application of the technique to a survey of a wide range of shampoo and skin cream types showed that levels of total N-nitroso compounds were less than 30 mug kg(-1) in < 90% of samples and, in many cases, less than 2.5 mug kg-1.

  19. 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.

  20. 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.

  1. Ultrasensitive and rapid screening of mercury(II) ions by dual labeling colorimetric method in aqueous samples and applications in mercury-poisoned animal tissues.

    PubMed

    Deng, Yi; Wang, Xin; Xue, Feng; Zheng, Lei; Liu, Jian; Yan, Feng; Xia, Fan; Chen, Wei

    2015-04-01

    Rapid and ultrasensitive detection of trace heavy metal mercury(II) ions (Hg(2+)) are of significant importance due to the induced serious risks for environment and human health. This presented article reports the gold nanoparticle-based dual labeling colorimetric method (Dual-COLO) for ultrasensitive and rapid detection of Hg(2+) using the specific thymine-Hg(2+)-thymine (T-Hg(2+)-T) as recognition system and the dual labeling strategy for signal amplification. Both qualitative and quantitative detections of Hg(2+) are achieved successfully in aqueous samples. More importantly, the achieved detection limit of 0.005 ng mL(-1) (0.025 nM) without any instruments is very competitive to other rapid detection methods even ICP-MS based methods. This Dual-COLO method is also applied directly for real water sample monitoring and, more importantly, applied in analysis of mercury poisoned animal tissues and body fluidic samples, indicating a potentially powerful and promising tool for environmental monitoring and food safety control.

  2. 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

  3. 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.

  4. 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.

  5. 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

  6. 4-(8-Quinolyl)amino-7-nitro-2,1,3-benzoxadiazole as a new colorimetric probe for rapid and visual detection of Hg2+

    NASA Astrophysics Data System (ADS)

    Wang, Ke; Yang, Lixue; Zhao, Chuan; Ma, Huimin

    2013-03-01

    4-Amino-7-nitro-2,1,3-benzoxadiazole (ANBD) usually serves as a scaffold for developing fluorescent probes. In this paper, however, ANBD has been used as a chromogenic unit to design a new colorimetric probe, 4-(8-quinolyl)amino-7-nitro-2,1,3-benzoxadiazole (1), for rapid and visual detection of Hg2+. The reaction of 1 with Hg2+ in NaH2PO4-Na2HPO4 buffer (pH 7.0) containing 70% (v/v) acetonitrile forms a 1:1 complex, accompanying a red shift of the absorption maximum from 482 nm to 557 nm and a distinct color change from orange to violet. Moreover the color reaction exhibits a high selectivity and sensitivity to Hg2+ only, instead of other common metal ions. This behavior may be ascribed to the formation of a specific 1-Hg2+ complex, which is supported by 1H NMR titration experiments. The present study is not only a supplement to the detection method of Hg2+, but also a merit to the chemistry of 4-amino-7-nitro-2,1,3-benzoxadiazole.

  7. 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

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

    2012-07-01

    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.

  8. 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…

  9. 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.

  10. 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.

  11. Rapid quantification of periodontitis-related bacteria using a novel modification of Invader PLUS technologies.

    PubMed

    Tadokoro, Kenichi; Yamaguchi, Toshikazu; Kawamura, Katsumi; Shimizu, Hajime; Egashira, Toru; Minabe, Masato; Yoshino, Toshiaki; Oguchi, Hirokazu

    2010-01-01

    The Invader PLUS technology is a sensitive, rapid method for the detection and quantification of nucleic acid. While the original technology is based on the amplification by polymerase chain reaction (PCR) of the target sequence followed by its detection using the Invader technology, the current modification allows simultaneous PCR amplification and Invader reaction. The PCR primers and the Invader probes are designed to operate at the same temperature. This allows simpler design and faster results. This technology has been applied for the quantification of six periodontitis-related bacteria (Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Toreponema denticola, Tannerella forsythensis and Fusobacterium nucleatum). Direct comparison of this modified Invader PLUS with real-time PCR demonstrated similar linear range. Furthermore, testing of 64 volunteers showed a good correlation between both technologies with correlation factors r2 spanning between 0.827 and 0.987. We demonstrated here that the proposed improvement of the Invader PLUS allows the detection and quantification of DNA sequences using a simple design and protocol that can be implemented in clinical testing. PMID:18718748

  12. 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

  13. 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.

  14. Rapid screening and quantification of sulfonate derivatives in white peony root by UHPLC-MS-MS.

    PubMed

    Yan, Zhixiang; Chen, Chen; Xie, Xiabing; Fu, Bo; Yang, Xinghao

    2012-02-01

    A rapid ultra-high-performance liquid chromatographic-tandem mass spectrometric (UHPLC-MS-MS) method has been developed for rapid screening and quantitative analysis of sulfonate derivatives (SDs) in commercial white peony root. Separation was performed on an Agilent Zorbax Eclipse Plus-C18 column by gradient elution with acetonitrile-0.1% (v/v) formic acid as the mobile phase. In-source fragmentation was used to generate the characteristic fragment ion at m/z 259 and to screen for nine SDs. Detection of these SDs was further performed in multiple reaction monitoring (MRM) mode to improve sensitivity and to quantify the two SDs paeoniflorin sulfonate and benzoylpaeoniflorin sulfonate. The method was validated for specificity, linearity, limits of detection and quantification, precision, accuracy, and matrix effects. Nine commercial white peony root samples were examined by use of this method, which revealed great variety in the paeoniflorin sulfonate and benzoylpaeoniflorin sulfonate content.

  15. Rapid diagnosis and quantification of herpes simplex virus with a green fluorescent protein reporter system.

    PubMed

    Kung, S H; Wang, Y C; Lin, C H; Kuo, R L; Liu, W T

    2000-11-01

    A genetically modified cell line (Vero-ICP10-EGFP) was constructed for detection of herpes simplex virus (HSV) by a simple, rapid and direct method. The cell line was developed by stable transfection of Vero cell with a plasmid encoding the green fluorescent protein (GFP) driven by the promoter of the HSV-2 ICP10 gene. As early as 6 h after infection with HSV, fluorescence-emitting cells can be observed under a fluorescence microscope. A single infected cell emitting fluorescence can be observed with soft agar overlay by inverted fluorescence microscopy. No induction of detectable fluorescence was seen following infections with human cytomegalovirus (HCMV), varicella zoster virus (VZV), coxsackievirus A16 and enterovirus 71. Analysis by flow cytometry also demonstrated that intensity of the triggered fluorescence is proportional to the titer of HSV inoculated. Taken together, this novel GFP reporter system could become a useful means for rapid detection and quantification of HSV in clinical specimens.

  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. Simple and portable magnetic immunoassay for rapid detection and sensitive quantification of plant viruses.

    PubMed

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

    2015-05-01

    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

  18. Rapid label-free visual assay for the detection and quantification of viral RNA using peptide nucleic acid (PNA) and gold nanoparticles (AuNPs).

    PubMed

    Joshi, Vinay G; Chindera, Kantaraja; Singh, Arvind Kumar; Sahoo, Aditya P; Dighe, Vikas D; Thakuria, Dimpal; Tiwari, Ashok K; Kumar, Satish

    2013-09-17

    A rapid label-free visual assay for the detection of viral RNA using peptide nucleic acid (PNA) probes and gold nanoparticles (AuNPs) is presented in this study. Diagnosis is a crucial step for the molecular surveillance of diseases, and a rapid visual test with high specificity could play a vital role in the management of viral diseases. In this assay, the specific agglomerative behavior of PNA with gold nanoparticles was manipulated by its complementation with viral RNA. The assay was able to detect 5-10 ng of viral RNA from various biological samples, such as allantoic fluids, cell culture fluids and vaccines, in 100 μl of test solution. The developed assay was more sensitive than a hemagglutination (HA) test, a routine platform test for the detection of Newcastle disease virus (NDV), and the developed assay was able to visually detect NDV with as little as 0.25 HA units of virus. In terms of the specificity, the test could discriminate single nucleotide differences in the target RNA and hence could provide visual viral genotyping/pathotyping. This observation was confirmed by pathotyping different known isolates of NDV. Further, the PNA-induced colorimetric changes in the presence of the target RNA at different RNA to PNA ratios yielded a standard curve with a linear coefficient of R(2)=0.990, which was comparable to the value of R(2)=0.995 from real-time PCR experiments with the same viral RNA. Therefore, the viral RNA in a given samples could be quantified using a simple visual spectrophotometer available in any clinical laboratory. This assay may find application in diagnostic assays for other RNA viruses, which are well known to undergo mutations, thus presenting challenges for their molecular surveillance, genotyping and quantification.

  19. Direct photothermal techniques for rapid quantification of total anthocyanin content in sour cherry cultivars.

    PubMed

    Dóka, Ottó; Ficzek, Gitta; Bicanic, Dane; Spruijt, Ruud; Luterotti, Svjetlana; Tóth, Magdolna; Buijnsters, Josephus Gerardus; Végvári, György

    2011-04-15

    The analytical performance of the newly proposed laser-based photoacoustic spectroscopy (PAS) and of optothermal window (OW) method for quantification of total anthocyanin concentration (TAC) in five sour cherry varieties is compared to that of the spectrophotometry (SP). High performance liquid chromatography (HPLC) was used to identify and quantify specific anthocyanins. Both, PAS and OW are direct methods that unlike SP and HPLC obviate the need for the extraction of analyte. The outcome of the study leads to the conclusion that PAS and OW are both suitable for quick screening of TAC in sour cherries. The correlation between the two methods and SP is linear with R(2)=0.9887 for PAS and R(2)=0.9918 for OW, respectively. Both methods are capable of the rapid determination of TAC in sour cherries without a need for a laborious sample pretreatment. PMID:21376955

  20. Rapid T2- and susceptometry-based CMRO2 quantification with interleaved TRUST (iTRUST).

    PubMed

    Rodgers, Zachary B; Englund, Erin K; Langham, Michael C; Magland, Jeremy F; Wehrli, Felix W

    2015-02-01

    Susceptometry-based oximetry (SBO) and T2-relaxation-under-spin-tagging (TRUST) are two promising methods for quantifying the cerebral metabolic rate of oxygen (CMRO2), a critical parameter of brain function. We present a combined method, interleaved TRUST (iTRUST), which achieves rapid, simultaneous quantification of both susceptometry- and T2-based CMRO2 via insertion of a flow-encoded, dual-echo gradient-recalled echo (OxFlow) module within the T1 recovery portion of the TRUST sequence. In addition to allowing direct comparison between SBO- and TRUST-derived venous oxygen saturation (Yv) values, iTRUST substantially improves TRUST temporal resolution for CMRO2 quantification and obviates the need for a separate blood flow measurement following TRUST acquisition. iTRUST was compared directly to TRUST and OxFlow alone in three resting subjects at baseline, exhibiting close agreement with the separate techniques and comparable precision. These baseline data as well as simulation results support the use of two instead of the traditional four T2 preparation times for T2 fitting, allowing simultaneous quantification of susceptometry- and T2-based Yv (and CMRO2) with three- and six-second temporal resolution, respectively. In 10 young healthy subjects, iTRUST was applied during a 5% CO2 gas mixture-breathing paradigm. T2-based Yv values were lower at baseline relative to susceptometry (62.3 ± 3.1 vs. 66.7 ± 5.1 %HbO2, P<0.05), but increased more in response to hypercapnia. As a result, T2-based CMRO2 decreased from 140.4 ± 9.7 to 120.0 ± 9.5 μMol/100g/min, a significant -14.6 ± 3.6% response (P < 0.0001), whereas susceptometry-based CMRO2 changed insignificantly from 123.4 ± 18.7 to 127.9 ± 25.7, a 3.3 ± 9.7% response (P = 0.31). These differing results are in accord with previous studies applying the parent OxFlow or TRUST sequences individually, thus supporting the reliability of iTRUST but also strongly suggesting that a systematic bias exists between the

  1. 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

  2. Rapid and simple colorimetric detection of Escherichia coli O157:H7 in apple juice using a novel recombinant bacteriophage-based Method.

    PubMed

    Hoang, Hoang A; Dien, Le T

    2015-01-01

    In this study, a bacteriophage-based method for the colorimetric detection of E. coli O157:H7 in apple juice was investigated. Firstly, a gene encoding Cytochrome c Peroxidase (CCP) chromogenic enzyme was inserted into a wild type PP01 phage genome to construct the recombinant PP01ccp phage that was used in the production of the chromogenic enzyme through specific infection into E. coli O157:H7. The method was then examined in the colorimetric detection of E. coli O157:H7 in broth, and the appearance of E. coli O157:H7 in broth was confirmed by the color change after a few minutes of the enzyme assay. Secondly, the method was investigated in the colorimetric detection of E. coli O157:H7 in apple juice. A low E. coli O157:H7 concentration as 1 CFU mL(-1) was detected in 15 h that was in a shorter time than in previous bioluminescence phage-based methods. Moreover, the method is much simpler compared to other previous phage-based methods since it enables detection without the need for expensive apparatus.

  3. A novel and rapid method for quantification of magnetic nanoparticle cell interactions using a desktop susceptometer

    NASA Astrophysics Data System (ADS)

    Ström, Valter; Hultenby, Kjell; Grüttner, Cordula; Teller, Joachim; Xu, Bo; Holgersson, Jan

    2004-05-01

    Activated endothelial cells (EC) are attractive prime targets for specific drug delivery using drug-carrying magnetic nanoparticles. In order to accomplish EC targeting, the interaction between magnetic particles and resting as well as activated endothelial cells must be characterized and quantified, because it will influence particle biodistribution, circulation half-time, and targeting efficacy. Here, we have quantified in vitro the interaction (adhesion/phagocytosis) between human endothelial cells and magnetite (Fe3O4) particles carrying different surface coatings with varying degrees of hydrophilicity and surface charge. Almost no adhesion was observed (about 1% or less) for three out of five particle types carrying plain dextran, carboxyl-substituted poly(ethylene glycol) and silica C18 coatings. In contrast, carboxyl-functionalized dextran and poly(ethylene glycol)-coated particles adhered or were phagocytosed to a considerable degree (58 and 26%, respectively). These clear and accurate results were obtained by measuring the magnetic response, i.e. magnetic susceptibility, from different fractions of the cell cultures as a means of determining the concentration of magnetic particles. Visible light and electron microscopy confirmed the magnetic quantification. To meet the need for a rapid yet sensitive instrument, we have developed a desktop magnetic susceptometer especially adapted for liquid samples or particles in a suspension. Despite its very high sensitivity, it is easy to operate and requires but a few seconds for a measurement. We also describe the construction and operation of this instrument.

  4. 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

  5. A Sensitive Assay for Rapid Detection and Quantification of Aphanomyces euteiches in Soil.

    PubMed

    Gangneux, Christophe; Cannesan, Marc-Antoine; Bressan, Mélanie; Castel, Lisa; Moussart, Anne; Vicré-Gibouin, Maïté; Driouich, Azeddine; Trinsoutrot-Gattin, Isabelle; Laval, Karine

    2014-10-01

    Aphanomyces euteiches is a widespread oomycete pathogen causing root rot in a wide range of leguminous crops. Losses can reach up to 100% for pea culture and there is currently no registered pesticide for its control. Crop management remains the most efficient tool to control root rot, and avoidance of infested soil seems to be the optimal solution. A test was developed to identify fields suitable for pea crops, consisting of the determination of the inoculum potential of soil using baiting plants. A new rapid, specific, and sensitive molecular method is described allowing the quantification of less than 10 oospores per gram of soil. This challenge is achieved by a real-time polymerase chain reaction procedure targeting internal transcribed spacer 1 from the ribosomal DNA operons. A preliminary study based on typical soils from northwestern France demonstrated that the A. euteiches oospore density in soil is related to the inoculum potential. Furthermore, this method has proved sensitive enough to accurately study the influence of biotic factors that may govern the actual emergence of root rot.

  6. 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

  7. 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.

  8. Selected reaction monitoring (SRM) mass spectrometry without isotope labeling can be used for rapid protein quantification.

    PubMed

    Zhi, Wenbo; Wang, Meiyao; She, Jin-Xiong

    2011-06-15

    The validation of putative biomarker candidates has become the major bottle-neck in protein biomarker development. Conventional immunoaffinity methods are limited by the availability of antibodies and kits. Here we demonstrate the feasibility of using selected reaction monitoring (SRM) without isotope labeling to achieve fast and reproducible quantification of serum proteins. The SRM/MRM assays for three standard serum proteins, including ceruloplasmin (CP), serum aymloid A (SAA) and sex hormone binding globulin (SHBG), have good linear ranges, generally 10(3) to 10(4) . There are almost perfect correlations between SRM intensities and the loaded peptide amounts (R(2) is usually ~0.99). Our data suggest that SRM/MRM is able to quantify proteins within the range of 0.2-2 fmol, which is comparable to the commercial ELISA/LUMINEX kits for these proteins. Excellent correlations between SRM/MRM and ELISA/LUMINEX assays were observed for SAA and SHBG (R(2)=0.928 and 0.851, respectively). However, the correlation between SRM/MRM and ELISA for CP is less desirable (R(2)=0.565). The reproducibility for SRM/MRM assays is generally very good but may depend on the proteins/peptides being analyzed (R(2)=0.931 and 0.882 for SAA and SHBG, and 0.723 for CP). The SRM/MRM assay without isotope labeling is a rapid and useful method for protein biomarker validation in a modest number of samples and is especially useful when other assays such as ELISA or LUMINEX are not available. PMID:21594933

  9. A novel fluorescence-based assay for the rapid detection and quantification of cellular deoxyribonucleoside triphosphates

    PubMed Central

    Wilson, Peter M.; LaBonte, Melissa J.; Russell, Jared; Louie, Stan; Ghobrial, Andrew A.; Ladner, Robert D.

    2011-01-01

    Current methods for measuring deoxyribonucleoside triphosphates (dNTPs) employ reagent and labor-intensive assays utilizing radioisotopes in DNA polymerase-based assays and/or chromatography-based approaches. We have developed a rapid and sensitive 96-well fluorescence-based assay to quantify cellular dNTPs utilizing a standard real-time PCR thermocycler. This assay relies on the principle that incorporation of a limiting dNTP is required for primer-extension and Taq polymerase-mediated 5–3′ exonuclease hydrolysis of a dual-quenched fluorophore-labeled probe resulting in fluorescence. The concentration of limiting dNTP is directly proportional to the fluorescence generated. The assay demonstrated excellent linearity (R2 > 0.99) and can be modified to detect between ∼0.5 and 100 pmol of dNTP. The limits of detection (LOD) and quantification (LOQ) for all dNTPs were defined as <0.77 and <1.3 pmol, respectively. The intra-assay and inter-assay variation coefficients were determined to be <4.6% and <10%, respectively with an accuracy of 100 ± 15% for all dNTPs. The assay quantified intracellular dNTPs with similar results obtained from a validated LC–MS/MS approach and successfully measured quantitative differences in dNTP pools in human cancer cells treated with inhibitors of thymidylate metabolism. This assay has important application in research that investigates the influence of pathological conditions or pharmacological agents on dNTP biosynthesis and regulation. PMID:21576234

  10. 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.

  11. 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.

  12. 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.

  13. Sensitive and rapid liquid chromatography/tandem mass spectrometric assay for the quantification of piperaquine in human plasma.

    PubMed

    Singhal, Puran; Gaur, Ashwani; Gautam, Anirudh; Varshney, Brijesh; Paliwal, Jyoti; Batra, Vijay

    2007-11-01

    A simple, sensitive and rapid liquid chromatography/tandem mass spectrometric (LC-MS/MS) method was developed and validated for quantification of piperaquine, an antimalarial drug, in human plasma using its structural analogue, piperazine bis chloroquinoline as internal standard (IS). The method involved a simple protein precipitation with methanol followed by rapid isocratic elution of analytes with 10mM ammonium acetate buffer/methanol/formic acid/ammonia solution (25/75/0.2/0.15, v/v) on Chromolith SpeedROD RP-18e reversed phase chromatographic column and quantification by mass spectrometry in the multiple reaction monitoring mode (MRM). The precursor to product ion transitions of m/z 535.3-->288.2 and m/z 409.1-->205.2 were used to measure the analyte and the IS, respectively. The assay exhibited a linear dynamic range of 1.0-250.2 ng/mL for piperaquine in plasma. The limit of detection (LOD) and lower limit of quantification (LLOQ) in plasma were 0.2 and 1.0 ng/mL, respectively. Acceptable precision and accuracy (+/-20% deviation for LLOQ standard and +/-15% deviation for other standards from the respective nominal concentration) were obtained for concentrations over the standard curve ranges. A run time of 2.5 min for a sample made it possible to achieve a throughput of more than 400 plasma samples analyzed per day. The validated method was successfully applied to analyze human plasma samples from phase-1 clinical studies. The mean pharmacokinetic parameters of piperaquine following 1000 mg oral dose: observed maximum plasma concentration (Cmax), time to maximum plasma concentration (Tmax) and elimination half-life (T1/2) were 46.1 ng/mL, 3.8h and 13 days, respectively. PMID:17923446

  14. 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

  15. A rapid HPLC method for indirect quantification of β-lactamase activity in milk.

    PubMed

    Zhou, Shuang; Wang, Dan; Zhao, Yunfeng; Wu, Yongning

    2015-04-01

    To circumvent the strictly regulated limits of antibiotics in milk, illegal addition of β-lactamase to lower the antibiotic levels in milk has been reported recently in China. Herein, we describe a fast, sensitive, and robust HPLC-UV method for the determination of β-lactamase activity in milk, based on an indirect quantification strategy. The test milk sample was mixed with a known amount of penicillin G, a specific substrate of β-lactamase. After incubation, an aliquot of the mixture was injected into the HPLC-UV system to quantify the remaining penicillin G in less than 10 min. Comparative analysis of the amount of penicillin G before and after incubation was used to indirectly deduce the activity of β-lactamase in the test sample. This method was successfully applied to milk products with different fat percentages, resulting in a detection limit of 0.6 U/mL. Good recoveries, ranging from 94 to 105%, were obtained from blank milk samples spiked with β-lactamase at levels of 2 to 50 U/mL, with relative standard deviations <6%. A good correlation was demonstrated between the HPLC method and the conventional culture-based assay. Using this method, the activity changes in β-lactamase during milk pasteurization, sterilization, and storage were investigated. The advantages of low-cost, accurate quantification and easily accessible instrumentation make the proposed method an ideal alternative for high-throughput routine analysis in the dairy industry.

  16. Rapid detection and quantification of specific proteins by immunodepletion and microfluidic separation

    PubMed Central

    Azadi, Glareh; Gustafson, Eric; Wessel, Gary M.; Tripathi, Anubhav

    2012-01-01

    Conventional immunoblotting techniques are labor intensive, time consuming and rely on the elution of target protein after depletion. Here we describe a new method for detection and quantification of proteins, independent of washing and elution. In this method, the target protein is first captured by immunodepletion with antibody coated microbeads. In the second step, both the supernatant after immunodepletion and the untreated protein sample are directly analyzed by microfluidic electrophoresis without further processing. Subsequently, the detection and quantification are performed comparing the electropherograms of these two samples. This method was tested using an Escherichia coli lysate with a FLAG-tagged protein and anti-FLAG magnetic beads. An incubation of as little as one minute was sufficient for detectable depletion (66%) by microchip electrophoresis. Longer incubation (up to 60 minutes) resulted in more depletion of the target band (82%). Our results show that only 19% of the target is recovered after elution from the beads. By eliminating multiple wash and elution steps, our method is faster, less labor intensive, and highly reproducible. Even in case of non-specific binding at low concentrations, the target protein can be easily identified. This work highlights the advantages of integrating immunodepletion techniques on a microfluidic platform. PMID:22539461

  17. Rapid quantification of dimethyl methylphosphonate from activated carbon particles by static headspace gas chromatography mass spectrometry.

    PubMed

    Mitchell, Brendan L; Billingsley, Brit G; Logue, Brian A

    2013-06-01

    Activated carbon (AC) particles are utilized as an adsorbent for binding hazardous vapors in protective equipment. The binding affinity and utilization of these AC particles should be known to ensure effective and efficient use. Therefore, a simple and effective method was developed for the quantification of the chemical warfare agent simulant, dimethyl methylphosphonate (DMMP), from AC particles. Static headspace gas chromatography mass-spectrometry with internal standard, DMMP-d6, was used to perform the analysis. The method produced a linear dynamic range of 2.48-620g DMMP/kg carbon and a detection limit of 1.24g DMMP/kg carbon. Furthermore, the method produced a coefficient of variation of less than 16% for all intra- and inter-assay analyses. The method provided a simple and effective procedure for quantifying DMMP from AC particles and was applied to the analysis of a DMMP-exposed AC protective respirator filter.

  18. 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

  19. Rapid quantification of dimethyl methylphosphonate from activated carbon particles by static headspace gas chromatography mass spectrometry.

    PubMed

    Mitchell, Brendan L; Billingsley, Brit G; Logue, Brian A

    2013-06-01

    Activated carbon (AC) particles are utilized as an adsorbent for binding hazardous vapors in protective equipment. The binding affinity and utilization of these AC particles should be known to ensure effective and efficient use. Therefore, a simple and effective method was developed for the quantification of the chemical warfare agent simulant, dimethyl methylphosphonate (DMMP), from AC particles. Static headspace gas chromatography mass-spectrometry with internal standard, DMMP-d6, was used to perform the analysis. The method produced a linear dynamic range of 2.48-620g DMMP/kg carbon and a detection limit of 1.24g DMMP/kg carbon. Furthermore, the method produced a coefficient of variation of less than 16% for all intra- and inter-assay analyses. The method provided a simple and effective procedure for quantifying DMMP from AC particles and was applied to the analysis of a DMMP-exposed AC protective respirator filter. PMID:23639122

  20. Rapid quantification of murine ABC mRNAs by real time reverse transcriptase-polymerase chain reaction.

    PubMed

    Su, Yan Ru; Linton, MacRae F; Fazio, Sergio

    2002-12-01

    Several ATP-binding cassette (ABC) transporters are critically involved in cholesterol and phospholipid efflux, reverse cholesterol transport, and play an important role in the development of atherosclerosis. Quantification of ABC mRNA is important in studying the regulation of cellular cholesterol homeostasis and mechanisms related to the pathogenesis of atherosclerosis. We have developed a one-step real time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) method for measuring mRNA levels of ABCA1, ABCG1, and ABCA2 in murine tissues using the TaqMan(TM) technology. It has significant methodological benefits when compared to classic Northern blotting or semi-quantitative RT-PCR analysis. Using this method, we found high expression levels of ABCA1 in liver and macrophages, and of ABCG1 in the brain and macrophages. The expression of ABCA1 and ABCG1 were further induced in macrophages loaded with acLDL. In contrast, ABCA2 was expressed exclusively in the brain with low expression levels in the macrophages. This method provides a rapid, highly sensitive, specific, and reproducible quantification of ABC mRNA, and can be performed with nanograms of total RNA sample, thus making it a superior method for studying the regulation of ABC transporters in cholesterol efflux and its role in the pathogenesis of atherosclerosis in murine models.

  1. 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.

  2. 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

  3. 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

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

    PubMed

    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

    2015-08-01

    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

  5. 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

  6. Adjustment of a rapid method for quantification of Fusarium spp. spore suspensions in plant pathology.

    PubMed

    Caligiore-Gei, Pablo F; Valdez, Jorge G

    2015-01-01

    The use of a Neubauer chamber is a broadly employed method when cell suspensions need to be quantified. However, this technique may take a long time and needs trained personnel. Spectrophotometry has proved to be a rapid, simple and accurate method to estimate the concentration of spore suspensions of isolates of the genus Fusarium. In this work we present a linear formula to relate absorbance measurements at 530nm with the number of microconidia/ml in a suspension.

  7. 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.

  8. 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

  9. 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

  10. 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

  11. 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

  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.

  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.

  14. Rapid detection and quantification of Cryptosporidium baileyi oocysts in feces and organs of chickens using a microscopic slide flotation method.

    PubMed

    Abbassi, H; Wyers, M; Cabaret, J; Naciri, M

    2000-03-01

    A simple semiquantitative microscopic slide flotation (MSF) method using modified Sheather's sugar solution (MSSS) is presented for the rapid detection and quantification of Cryptosporidium baileyi oocysts in the feces and mucosal and/or organ scrapings of chickens. Oocyst shedding was evaluated by examination of the surface of coverslips, and the average quantitative score (0-5) recorded for 10 microscopic fields (magnification x250) is reported. The equivalence between these scores and the actual number of oocysts counted per gram of feces was assessed (rs = 0.89; P < 0.001). The applicability of this method was tested by comparison of the kinetics of oocyst shedding in feces of inoculated chickens with those reported by other authors working under similar conditions. In organs the MSF method was compared to histology. Fewer false-negative results were obtained using MSF versus the histology method. The MSF method was particularly more efficient in tracheae with low levels of infection and in the lungs, regardless of the level of infection. The MSF method was also very efficient in detecting oocysts in air sacs from chickens with aerosacculitis. It provides a specific and sufficiently sensitive, simple, rapid, reliable, and low-cost means of diagnosing C. baileyi in the feces and organs of chickens. This method can be used in the routine diagnosis of cryptosporidia in chickens, and it could be extended to other avian species and used in epidemiology studies to evaluate the prevalence of cryptosporidiosis in fowl.

  15. A real-time PCR assay for rapid detection and quantification of Exserohilum rostratum, a causative pathogen of fungal meningitis associated with injection of contaminated methylprednisolone.

    PubMed

    Zhao, Yanan; Petraitiene, Ruta; Walsh, Thomas J; Perlin, David S

    2013-03-01

    A species-specific molecular beacon real-time PCR assay was developed for rapid diagnosis of Exserohilum rostratum infection. As low as 100 fg of E. rostratum DNA can be reliably detected in the presence of 50 ng of human DNA, with a dynamic linear quantification range from 20 ng to 200 fg.

  16. 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.

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

    PubMed

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

    1986-02-01

    We designed, constructed, and evaluated a new device to characterize the human carotid baroreceptor-cardiac reflex response relation rapidly. We designed this system for study of reflex responses of astronauts before, during, and after space travel. The system comprises a new tightly sealing silicone rubber neck chamber, a stepping motor-driven electro-deposited 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.

  18. Rapid quantitative colourimetric tests for trinitrotoluene (TNT) in soil.

    PubMed

    Choodum, Aree; Kanatharana, Proespichaya; Wongniramaikul, Worawit; NicDaeid, Niamh

    2012-10-10

    A rapid quantitative colourimetric test was developed for the determination of 2,4,6-Trinitrotoluene (TNT) in soil. Quantification was achieved using an innovative application of photography where the relationship between the Red Green Blue (RGB) value and concentration of colorimetric product was exploited. The wide linear range (1-50 mg/L) and low detection limit (0.73±0.01 mg/L to 3.5±0.4 mg/L) achieved was comparable with spectrophotometric quantification methods. Quantification was validated both with known concentration standards and spiked soil extracts confirming the accuracy of the technique. The results demonstrate a powerful technique with the capacity for development as a rapid on site field test for the analysis of explosives.

  19. 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.

  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 fluorescent reporter quantification by leaf disc analysis and its application in plant-virus studies

    PubMed Central

    2014-01-01

    Background Fluorescent proteins are extraordinary tools for biology studies due to their versatility; they are used extensively to improve comprehension of plant-microbe interactions. The viral infection process can easily be tracked and imaged in a plant with fluorescent protein-tagged viruses. In plants, fluorescent protein genes are among the most commonly used reporters in transient RNA silencing and heterologous protein expression assays. Fluorescence intensity is used to quantify fluorescent protein accumulation by image analysis or spectroscopy of protein extracts; however, these methods might not be suitable for medium- to large-scale comparisons. Results We report that laser scanners, used routinely in proteomic studies, are suitable for quantitative imaging of plant leaves that express different fluorescent protein pairs. We developed a microtiter plate fluorescence spectroscopy method for direct quantitative comparison of fluorescent protein accumulation in intact leaf discs. We used this technique to measure a fluorescent reporter in a transient RNA silencing suppression assay, and also to monitor early amplification dynamics of a fluorescent protein-labeled potyvirus. Conclusions Laser scanners allow dual-color fluorescence imaging of leaf samples, which might not be acquired in standard stereomicroscope devices. Fluorescence microtiter plate analysis of intact leaf discs can be used for rapid, accurate quantitative comparison of fluorescent protein accumulation. PMID:25053970

  2. 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

  3. 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.

  4. Rapid quantification of adenosine cyclic 3',5'-monophosphate by competitive enzyme-linked immunosorbent assay.

    PubMed

    Hsieh, M S; Jap, T S; Chiang, H

    1993-01-01

    A reliable and rapid enzyme-linked immunosorbent assay (ELISA) for cyclic AMP determination is described. Succinyl cyclic AMP, coupled to human albumin, was injected into rabbit to elicit antibodies to cyclic nucleotide hapten. Succinyl cyclic nucleotide to human albumin as immunogen or the cyclic AMP to porcine thyroglobulin as coating antigen was conjugated by a carbodiimide coupling procedure. The latter conjugate, captured to microplate with coating buffer and blocked with 0.8% gelatin for 30 minutes, was bound to antibody in inverse proportion to free cyclic AMP in a sample or standard. Bound antibody was then quantified with horseradish peroxidase-labelled goat antirabbit immunoglobulin and ABTS (2, 2'-Azinobis (3-ethylbenzthiazolinesulfonic Acid). Our results showed that concentration of both standard and sample cyclic AMP could be measured as low as 2.5 fmol/well (0.05 pmol/ml). The intra- and inter-assay coefficients of variation for samples were 6.0-8.0% and 8.9-9.5%, respectively. In addition, there was no cross-reaction of the antisera with ADP, ATP, 5'-AMP or cyclic GMP. Short period of incubation at room temperature seems as good as long period of incubation at 4 degrees C. The biological study demonstrated a consistency between increase in platelet-cyclic AMP generation after prostaglandin E1 stimulation and its biological effects. Our approach to ELISA is validated by showing agreement in levels, obtained in parallel by ELISA and RIA, of cyclic AMP content in extracts of prostaglandin E1-stimulated platelet cells.

  5. 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.

  6. 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.

  7. 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

  8. Rapid speciation and quantification of selenium compounds by HPLC-ICP MS using multiple standards labelled with different isotopes.

    PubMed

    Ohta, Yuki; Suzuki, Noriyuki; Kobayashi, Yayoi; Hirano, Seishiro

    2011-09-01

    Speciation analysis using high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP MS) is now commonly used to investigate metabolic and toxicological aspects of some metals and metalloids. We have developed a rapid method for simultaneous identification and quantification of metabolites of selenium (Se) compounds using multiple standards labelled with different isotopes. A mixture of the labelled standards was spiked in a selenised garlic extract and the sample was subjected to speciation analysis by HPLC-ICP MS. The selenised garlic contains γ-glutamyl-methylselenocysteine, methylselenocysteine, and selenomethionine and the concentrations of those Se compounds were 723.8, 414.8, and 310.7 ng Se ml(-1), respectively. The isotopically labelled standards were also applied to the speciation of Se in rat urine. Selenate, methylselenonic acid, selenosugar, and trimethyselenium ions were found to be excreted by the present speciation procedure. Multiple standards labelled with different stable isotopes enable high-throughput identification and quantitative measurements of Se metabolites.

  9. 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).

  10. Simple, rapid and sensitive assay method for simultaneous quantification of urinary nicotine and cotinine using gas chromatography-mass spectrometry.

    PubMed

    Man, Che Nin; Gam, Lay-Harn; Ismail, Syazwani; Lajis, Razak; Awang, Rahmat

    2006-12-01

    Nicotine is a major addictive compound in cigarette. Its smoke is rapidly and extensively metabolized to several metabolites in human. Cotinine as a major metabolite of nicotine is commonly used as a biomarker to determine active and passive smokers. Cotinine has a longer half-life ( approximately 20 h) compared to nicotine ( approximately 2h). A simple, sensitive, rapid and high throughput GC-MS method was developed for simultaneous quantification of urinary nicotine and cotinine in passive and active smokers. In the sample preparation method, the analytes and internal standard were first basified and followed by liquid-liquid extraction. Upon completion, anhydrous sodium sulphate was added to the solvent mixture to trap moistures. The clear extract obtained was directly injected into GC-MS, operating under selective ion monitoring (SIM) mode. Calibration curves in the range of 0.5-5000 ng/mL of the analytes in urine matrix were established with linear correlation coefficients (r(2)) greater than 0.997. The limit of detection for both nicotine and cotinine were 0.20 ng/mL. The mean recoveries for nicotine and cotinine were 93.0 and 100.4%, respectively. The within- and between-assay accuracies were between 2.1 and 7.9% for nicotine and between 0.7 and 11.1% for cotinine. Within- and between-assay precisions of 3.3-9.5% for nicotine and 3.4-9.8% for cotinine were also achieved. The method can be used in routine assessment and monitoring of active smoking and exposure to environmental tobacco smoke. The applicability of the assay was demonstrated in a small-scale comparison study between smokers and non-smokers. PMID:16908224

  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. 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.

  14. Colorimetric determination of ferrous-ferric ratio in glass

    SciTech Connect

    Baumann, E W

    1987-02-20

    This memorandum describes a simple, rapid colorimetric procedure that can be used for analysis of vitrified melter feed. This method can replace the Moessbauer spectroscopy method for routine DWPF process control analysis. A colorimetric method has been developed that is recommended as a replacement for Moessbauer spectroscopy for process control measurements of the Fe(II)/Fe(III) ratio of melter feed for the DWPF analytical laboratory. The results agree satisfactorily with Moessbauer values. The method is suitable for remote operation and has several advantages. The total time involved is less than 30 minutes. Quantitative measurements of samples, reagents, and dilutions are unnecessary because only the ratio value is required and both Fe(II) and total Fe are measured on the same solution. Small samples (<3 mg total Fe) are required, minimizing radiation exposure, because of the high sensitivity of the colorimetric method. Relative precision and relative accuracy are about {plus minus}5%.

  15. 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

  16. 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

  17. 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

  18. 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.

  19. Molecular diagnosis of α-thalassemias by the colorimetric nanogold.

    PubMed

    Chomean, Sirinart; Wangmaung, Nantawan; Sritongkham, Pornpimol; Promptmas, Chamras; Mas-Oodi, Sumana; Tanyong, Dalina; Ittarat, Wanida

    2014-02-21

    A new application of gold nanoparticles (AuNPs) as a colorimetric method for gene detection of α-thalassemia 1 (SEA deletion) is reported here for the first time. This technique is based on color changes from salt-induced aggregation of un-hybridized nanogold probes after hybridization with the target DNA. Specific DNA probes were synthesized, thiol modified and conjugated on the surface of AuNPs. The target DNA was amplified and hybridized with the AuNPs-immobilized probe. Salt solution (NaCl) was added to induce aggregation of the un-hybridized nanogold probes. The color changes were visualized either by the naked eye or by UV-vis spectrophotometry at 520 nm. By this nanogold colorimetric method samples carrying normal α-globin genes could be successfully identified from samples carrying α-globin genes causing α-thalassemia 1 (SEA deletion), either as a carrier or disease form. Results demonstrated that the new colorimetric nanogold method is a definite gene diagnosis of α-thalassemia. It is accurate, simple, rapid, specific, sensitive, and cost effective. It is also a promising point-of-care testing (POCT) method for thalassemias and other genetic disorders. The new colorimetric nanogold is a method of choice for areas where access to sophisticated molecular diagnosis is limited.

  20. Colorimetric detection of mercury ions based on plasmonic nanoparticles.

    PubMed

    Du, Jianjun; Jiang, Lin; Shao, Qi; Liu, Xiaogang; Marks, Robert S; Ma, Jan; Chen, Xiaodong

    2013-05-27

    The development of rapid, specific, cost-effective, and robust tools in monitoring Hg(2+) levels in both environmental and biological samples is of utmost importance due to the severe mercury toxicity to humans. A number of techniques exist, but the colorimetric assay, which is reviewed herein, is shown to be a possible tool in monitoring the level of mercury. These assays allow transforming target sensing events into color changes, which have applicable potential for in-the-field application through naked-eye detection. Specifically, plasmonic nanoparticle-based colorimetric assay exhibits a much better propensity for identifying various targets in terms of sensitivity, solubility, and stability compared to commonly used organic chromophores. In this review, recent progress in the development of gold nanoparticle-based colorimetric assays for Hg(2+) is summarized, with a particular emphasis on examples of functionalized gold nanoparticle systems with oligonucleotides, oligopeptides, and functional molecules. Besides highlighting the current design principle for plasmonic nanoparticle-based colorimetric probes, the discussions on challenges and the prospect of next-generation probes for in-the-field applications are also presented.

  1. 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.

  2. A comparison of two colorimetric assays, based upon Lowry and Bradford techniques, to estimate total protein in soil extracts.

    PubMed

    Redmile-Gordon, M A; Armenise, E; White, R P; Hirsch, P R; Goulding, K W T

    2013-12-01

    Soil extracts usually contain large quantities of dissolved humified organic material, typically reflected by high polyphenolic content. Since polyphenols seriously confound quantification of extracted protein, minimising this interference is important to ensure measurements are representative. Although the Bradford colorimetric assay is used routinely in soil science for rapid quantification protein in soil-extracts, it has several limitations. We therefore investigated an alternative colorimetric technique based on the Lowry assay (frequently used to measure protein and humic substances as distinct pools in microbial biofilms). The accuracies of both the Bradford assay and a modified Lowry microplate method were compared in factorial combination. Protein was quantified in soil-extracts (extracted with citrate), including standard additions of model protein (BSA) and polyphenol (Sigma H1675-2). Using the Lowry microplate assay described, no interfering effects of citrate were detected even with concentrations up to 5 times greater than are typically used to extract soil protein. Moreover, the Bradford assay was found to be highly susceptible to two simultaneous and confounding artefacts: 1) the colour development due to added protein was greatly inhibited by polyphenol concentration, and 2) substantial colour development was caused directly by the polyphenol addition. In contrast, the Lowry method enabled distinction between colour development from protein and non-protein origin, providing a more accurate quantitative analysis. These results suggest that the modified-Lowry method is a more suitable measure of extract protein (defined by standard equivalents) because it is less confounded by the high polyphenolic content which is so typical of soil extracts.

  3. 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.

    PubMed

    Xie, G; Barcelona, M J; Fang, J

    1999-05-01

    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 currently used methods (GC/MS, conventional EPA 418.1, and a rapid field method PetroFLAG) were performed to quantify the TPH content in samples collected from a site contaminated by transformer oil. To standardize the method and improve the comparability of TPH data, crucial GC-based quantification issues were examined, e.g., quantification based on internal standards (ISTD) vs external standards (ESTD), single vs multiple ISTD, and various area integration approaches. The interpretation of hydrocarbon chromatographic results was examined in the context of field samples. The performance of the GC/MS method was compared with those of EPA 418.1 and PetroFLAG. As a result, it was observed that the ISTD quantification method was preferred to the ESTD method, multiple ISTD might be better than single ISTD, and three different area integration approaches did not have a significant effect on TPH results. Evaluation of the chromatograms between a reference sample and three unknown samples showed that the extent of contamination varied appreciably with sample depth. It was also found that there existed a good positive correlation between GC/MS and both EPA 418.1 and PetroFLAG, and that EPA 418.1 produced the higher overall estimate while GC/MS and PetroFLAG resulted in lower, more statistically comparable TPH values.

  4. 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.

  5. 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.

  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. Rapid, sensitive and selective HPLC-MS/MS method for the quantification of topically applied besifloxacin in rabbit plasma and ocular tissues: Application to a pharmacokinetic study.

    PubMed

    Gu, Xiao-Fei; Mao, Bai-Yang; Xia, Min; Yang, Yang; Zhang, Jia-Li; Yang, Da-Song; Wu, Wei-Xin; Du, Ying-Xiang; Di, Bin; Su, Meng-Xiang

    2016-01-01

    Besifloxacin is a fourth-generation broad-spectrum fluoroquinolone registered for the topical treatment of bacterial conjunctivitis. In this study, a rapid, sensitive and selective liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for quantification of besifloxacin in rabbit plasma and ocular tissues using nateglinide as the internal standard (IS). The analyte and IS were separated on a Sepax GP-Phenyl column by isocratic elution with methanol-acetonitrile-5 mM ammonium formate-formic acid (29:55:16:0.1, v/v/v/v) as the mobile phase at a flow rate of 1.2 mL/min, and the total run time was 3.0 min. An electrospray ionization (ESI) source was applied and operated in the positive ion mode; multiple reaction monitoring (MRM) mode was used for quantification, and the monitored transitions were 394.2→377.1 for besifloxacin and m/z 318.3→166.1 for the IS. The calibration curve was linear over the range of 0.103-206 ng/mL for plasma and 2.06-2060 ng/mL for tears, aqueous humor, conjunctiva and cornea with correlation coefficient (r) greater than 0.99. The lower limit of quantification (LLOQ) for besifloxacin was 0.103 ng/mL for plasma and 2.06 ng/mL for other ocular tissues with good accuracy and precision. Intra- and inter-batch precision were both lower than 15% and accuracy ranged from 85% to 115% at all QC levels. The method was successfully applied to the pharmacokinetic study of besifloxacin in rabbit plasma and ocular tissues after single and multiple topical administrations. PMID:26340560

  8. 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.

  9. A rapid HPLC with evaporative light scattering method for quantification of forskolin in multi-herbal weight-loss solid oral dosage forms.

    PubMed

    Virgona, N; Taki, Y; Umegaki, K

    2010-05-01

    A rapid reverse-phase HPLC method with evaporative light scattering detection (ELSD) was developed for the determination of forskolin in weight loss multi-herbals products. The analysis was performed by water-acetonitrile gradient elution at a temperature of 40 degrees C and a flow rate of 1.0 mL/min. The evaporator tube temperature of ELSD was set at 35 degrees C, and with the nebulizing gas flow-rate (pressure) of 3.0 bar. The method was validated for linearity, accuracy, precision and limits of detection (LOD) and quantification (LOQ). Good linear relationships were obtained with correlation coefficients exceeding 0.9995. The average recovery of forskolin ranged from 99.4% to 100.4% with RSDs below 3%. The percent relative standard deviations (%RSD) of intra- and inter-day precision varied by less than 2.1%. LOD and LOQ were 0.95 microg/ml and 3.21 microg/ml, respectively. The validated ELSD method permits a shorter determination time without compromising accuracy and demonstrates that it can be used for quantification of forskolin incorporated in multi-herbal solid oral dosage forms. PMID:20503921

  10. 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

  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

    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

  12. 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

  13. 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.

  14. Colorimetric detection of mercury species based on functionalized gold nanoparticles.

    PubMed

    Chen, Ling; Li, Jinhua; Chen, Lingxin

    2014-09-24

    The speciation analysis of heavy metal pollutants is very important because different species induce different toxicological effects. Nanomaterial-assisted optical sensors have achieved rapid developments, displaying wide applications to heavy metal ions but few to metal speciation analysis. In this work, a novel colorimetric nanosensor strategy for mercury speciation was proposed for the first time, based on the analyte-induced aggregation of gold nanoparticles (Au NPs) with the assistance of a thiol-containing ligand of diethyldithiocarbamate (DDTC). Upon the addition of mercury species, because Hg-DDTC was more stable than Cu-DDTC, a place-displacement between Hg species and Cu(2+) would occur, and thereby the functionalized Au NPs would aggregate, resulting in a color change. Moreover, by virtue of the masking effect of ethylenediaminetetraacetic acid (EDTA), the nanosensor could readily discriminate organic mercury and inorganic mercury (Hg(2+)), and it is thus anticipated to shed some light on the colorimetric sensing of organic mercury. So, a direct, simple colorimetric assay for selective determination of Hg species was obtained, presenting high detectability, such as up to 10 nM for Hg(2+) and 15 nM for methylmercury. Meanwhile, the strategy offered excellent selectivity toward mercury species against other metal ions. The simple, rapid, and sensitive label-free colorimetric sensor for the determination of Hg species provided an attractive alternative to conventional methods, which usually involve sophisticated instruments, complicated processes, and long periods of time. More importantly, by using mercury as a model, an excellent nanomaterial-based optical sensing platform can be developed for speciation analysis of trace heavy metals, which can lead to nanomaterials stability change through smart functionalization and reasonable interactions.

  15. 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).

  16. Simple colorimetric method determines uranium in tissue

    NASA Technical Reports Server (NTRS)

    Doran, D.; Frigerio, N. A.

    1967-01-01

    Simple colorimetric micromethod determines concentrations of uranium in tissue. The method involves dry ashing organic extraction, and colorimetric determination of uranyl ferrocyanide. This uranium determination technique could be used in agricultural research, tracer studies, testing of food products, or medical research.

  17. Rapid authentication of coffee blends and quantification of 16-O-methylcafestol in roasted coffee beans by nuclear magnetic resonance.

    PubMed

    Schievano, Elisabetta; Finotello, Claudia; De Angelis, Elisabetta; Mammi, Stefano; Navarini, Luciano

    2014-12-24

    Roasted coffee is subject to commercial frauds, because the high-quality Coffea arabica species, described as "100% Arabica" or "Highland coffee", is often mixed with the less expensive Coffea canephora var. Robusta. The quantification of 16-O-methylcafestol (16-OMC) is useful to monitor the authenticity of the products as well as the Robusta content in blends. The German standard method DIN 10779 is used in the determination of 16-OMC in roasted coffee beans to detect C. canephora in blends, but it is laborious and time-consuming. Here, we introduce a new method that provides a quantitative determination of esterified 16-OMC directly in coffee extracts by means of high-resolution proton nuclear magnetic resonance spectroscopy. Limit of detection and limit of quantitation were 5 and 20 mg/kg, respectively, which are adequate to detect the presence of Robusta at percentages lower than 0.9%. The proposed method is much faster, more sensitive, and much more reproducible than the DIN standard method.

  18. Rapid method for quantification of nine sulfonamides in bovine milk using HPLC/MS/MS and without using SPE.

    PubMed

    Nebot, Carolina; Regal, Patricia; Miranda, Jose Manuel; Fente, Cristina; Cepeda, Alberto

    2013-12-01

    Sulfonamides are antimicrobial agents widely employed in animal production and their residues in food could be an important risk to human health. In the dairy industry, large quantities of milk are monitored daily for the presence of sulfonamides. A simple and low-cost extraction protocol followed by a liquid chromatographic-tandem mass spectrometry method was developed for the simultaneous detection of nine sulfonamides in whole milk. The method was validated at the maximum residue limits established by European legislation. The limits of quantification obtained for most sulfonamides were between 12.5 and 25 μg kg(-1), detection capabilities ranged from 116 to 145 μg kg(-1), and recoveries, at 100 μg kg(-1), were greater than 89±12.5%. The method was employed to analyse 100 raw whole bovine milk samples collected from dairy farms in the northwest region of Spain. All of the samples were found to be compliant, but two were positive; one for sulfadiazine and the other for sulfamethoxipyridazine.

  19. 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

  20. 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.

  1. Colorimetric evaluation of display performance

    NASA Astrophysics Data System (ADS)

    Kosmowski, Bogdan B.

    2001-08-01

    The development of information techniques, using new technologies, physical phenomena and coding schemes, enables new application areas to be benefited form the introduction of displays. The full utilization of the visual perception of a human operator, requires the color coding process to be implemented. The evolution of displays, from achromatic (B&W) and monochromatic, to multicolor and full-color, enhances the possibilities of information coding, creating however a need for the quantitative methods of display parameter assessment. Quantitative assessment of color displays, restricted to photometric measurements of their parameters, is an estimate leading to considerable errors. Therefore, the measurements of a display's color properties have to be based on spectral measurements of the display and its elements. The quantitative assessment of the display system parameters should be made using colorimetric systems like CIE1931, CIE1976 LAB or LUV. In the paper, the constraints on the measurement method selection for the color display evaluation are discussed and the relations between their qualitative assessment and the ergonomic conditions of their application are also presented. The paper presents the examples of using LUV colorimetric system and color difference (Delta) E in the optimization of color liquid crystal displays.

  2. Development of a real-time PCR method (Taqman) for rapid identification and quantification of Prorocentrum donghaiense

    NASA Astrophysics Data System (ADS)

    Yuan, Jian; Mi, Tiezhu; Zhen, Yu; Yu, Zhigang

    2012-09-01

    Prorocentrum donghaiense is a dinoflagellate that is widely distributed in the East China Sea and has become increasingly involved in Harmful Algal Blooms (HABs). Therefore, it is necessary to study this dinoflagellate to monitor HABs. In this study, 13 pairs of primers specific to P. donghaiense (within its internal transcribed spacer (ITS) regions) were designed for SYBR Green I real-time PCR. As the SYBR Green I real-time PCR could not identify P. donghaiense in a specific manner, a Taqman real-time PCR method was developed by designing a set of specific primers and a Taqman probe. A 10-fold serial dilution of recombinant plasmid containing ITS regions of P. donghaiense was prepared as standard samples and the standard curve was established. Additionally, we quantified the genomic DNA in P. donghaiense cells and utilized this DNA to prepare another 10-fold serial dilution of standard sample and accordingly set up the standard curve. The mathematic correlation between the cell number and its corresponding plasmid copy number was also established. In order to test the efficiency of the real-time PCR method, laboratory samples and P. donghaiense HAB field samples were employed for identification and quantitative analysis. As to laboratory samples, as few as 102 cells of P. donghaiense could be quantified precisely utilizing both centrifugation and filtration techniques. The quantification results from field samples by real-time PCR were highly similar to those by light microscopy. In conclusion, the real-time PCR could be applied to identify and quantify P. donghaiense in HABs.

  3. 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

  4. 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

  5. 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.

  6. Colorimetric determination of inorganic iodine in fortified culinary products.

    PubMed

    Perring, I; Basic-Dvorzak, M; Andrey, D

    2001-07-01

    The presented colorimetric procedure only requires simple laboratory equipment and is suitable as a routine procedure for checking concentrations of iodine in fortified culinary products. The Moxon and Dixon colorimetric procedure for iodine determination has been optimised for the determination of iodide and iodate in fortified culinary products, always containing high salt levels. The high sensitivity of the method permits a high dilution of the product solutions, thus reducing interferences from the inherent colour of the products. The calibration is linear in the range from 0 to 12 microg L(-1) of iodine with R2 > 0.99. A series of commercial culinary products were used to validate the method. Recoveries of iodine, added as iodide and/or iodate, were generally in the range 100+/-10%. High concentrations of chloride are essential to obtain a complete recovery of iodate. Limit of quantification was estimated to be 2 mg kg(-1) of product, based on 2-3 g of product. Concentrations of iodine determined with this method were similar to those obtained by an ICP-MS procedure.

  7. Rapid quantification of viable Campylobacter bacteria on chicken carcasses, using real-time PCR and propidium monoazide treatment, as a tool for quantitative risk assessment.

    PubMed

    Josefsen, M H; Löfström, C; Hansen, T B; Christensen, L S; Olsen, J E; Hoorfar, J

    2010-08-01

    A number of intervention strategies against Campylobacter-contaminated poultry focus on postslaughter reduction of the number of cells, emphasizing the need for rapid and reliable quantitative detection of only viable Campylobacter bacteria. We present a new and rapid quantitative approach to the enumeration of food-borne Campylobacter bacteria that combines real-time quantitative PCR (Q-PCR) with simple propidium monoazide (PMA) sample treatment. In less than 3 h, this method generates a signal from only viable and viable but nonculturable (VBNC) Campylobacter bacteria with an intact membrane. The method's performance was evaluated by assessing the contributions to variability by individual chicken carcass rinse matrices, species of Campylobacter, and differences in efficiency of DNA extraction with differing cell inputs. The method was compared with culture-based enumeration on 50 naturally infected chickens. The cell contents correlated with cycle threshold (C(T)) values (R(2) = 0.993), with a quantification range of 1 x 10(2) to 1 x 10(7) CFU/ml. The correlation between the Campylobacter counts obtained by PMA-PCR and culture on naturally contaminated chickens was high (R(2) = 0.844). The amplification efficiency of the Q-PCR method was not affected by the chicken rinse matrix or by the species of Campylobacter. No Q-PCR signals were obtained from artificially inoculated chicken rinse when PMA sample treatment was applied. In conclusion, this study presents a rapid tool for producing reliable quantitative data on viable Campylobacter bacteria in chicken carcass rinse. The proposed method does not detect DNA from dead Campylobacter bacteria but recognizes the infectious potential of the VBNC state and is thereby able to assess the effect of control strategies and provide trustworthy data for risk assessment.

  8. Direct, simultaneous quantification of fructooligosaccharides by FT-MIR ATR spectroscopy and chemometrics for rapid identification of superior, engineered β-fructofuranosidases.

    PubMed

    Trollope, Kim M; Volschenk, Heinrich; Görgens, Johann F; Bro, Rasmus; Nieuwoudt, Hélène H

    2015-02-01

    Fructooligosaccharides (FOS) are popular components of functional foods produced by the enzymatic transfer of fructose units to sucrose. Improving β-fructofuranosidase traits by protein engineering is restricted by the absence of a rapid, direct screening method for the fructooligosaccharide products produced by enzyme variants. The use of standard high-performance liquid chromatography (HPLC) methods involves time-consuming sample preparation and chromatographic and data analysis steps. To overcome these limitations, this work presents a rapid method for screening β-fructofuranosidase variant libraries using Fourier transform mid-infrared attenuated total reflectance (FT-MIR ATR) spectroscopy and calibration using partial least squares (PLS) regression. The method offers notable improvements in terms of sample analysis times and cost, with the added benefit of the absence of toxic eluents. Wavenumber interval selection methods were tested to develop optimised PLS regression models that were successfully applied to quantify of glucose, fructose, sucrose, 1-kestose and nystose, the substrates and products of β-fructofuranosidase activity. To the best of our knowledge, this is the first report on the use of infrared spectroscopy and PLS calibration for the quantification of 1-kestose and nystose. Independent test set-validated results indicated that optimal wavenumber selection by interval PLS (iPLS) served to provide the best models for all sugars, bar glucose. Application of this screening method will facilitate the engineering of β-fructofuranosidases and other glycosyltransferase enzymes by random mutagenesis strategies, as it provides, for the first time, a rapid, direct assay for transferase products that may be adapted to a high-throughput set-up. PMID:25542584

  9. Rapid Quantification of Cell Viability and Apoptosis in B-Cell Lymphoma Cultures Using Cyanine SYTO Probes

    PubMed Central

    Wlodkowic, Donald; Skommer, Joanna; Darzynkiewicz, Zbigniew

    2011-01-01

    The gross majority of classical apoptotic hallmarks can be rapidly examined by multiparameter flow cytometry. As a result, cytometry became a technology of choice in diverse studies of cellular demise. In this context, a novel class of substituted unsymmetrical cyanine SYTO probes has recently become commercially available. Derived from thiazole orange, SYTO display low intrinsic fluorescence, with strong enhancement upon binding to DNA and/or RNA. Broad selection of excitation/emission spectra has recently driven implementation of SYTO dyes in polychromatic protocols with the detection of apoptosis being one of the most prominent applications In this chapter, we outline a handful of commonly used protocols for the assessment of apoptotic events using selected SYTO probes (SYTO 16, 62, 80) in conjunction with common plasma membrane permeability markers (PI, YO-PRO 1, 7-AAD). PMID:21468970

  10. 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

  11. 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

  12. 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.

  13. 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 Central

    Ingram, David T.; Lamichhane, Chinta M.; Rollins, David M.; Carr, Lewis E.; Mallinson, Edward T.; Joseph, Sam W.

    1998-01-01

    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°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 traditional

  14. Rapid quantification of histamine in human psoriatic plaques using microdialysis and ultra high performance liquid chromatography with fluorescence detection.

    PubMed

    Guihen, Elizabeth; Ho, Wen Lyn; Hogan, Anna-Marie; O'Connell, Marie-Louise; Leahy, Martin J; Ramsay, Bart; O'Connor, William T

    2012-01-01

    Psoriasis is a chronic skin disease resulting from abnormal immune function and is characterized by the presence of scaly psoriatic plaques which are areas of inflammation and excessive skin production. The psoriatic plaques contain mast cells which are increased in number in the uppermost dermis of the psoriatic lesion and which may play a role in the initiation and maintenance of the lesion. These processes are thought to be mediated via the local release of histamine along with other mediators from the mast cells; however their precise role still remains a mystery. Our study involved the development of a rapid and ultra-sensitive liquid chromatographic method for the separation and detection of histamine. To this end a state-of-the-art ultra high pressure liquid chromatography (UHPLC) system incorporating the latest technology in fluorescence detection system was employed which allowed for the rapid and reliable trace level detection of histamine in human derived microdialysate samples. This new reverse phase method utilized a sub-two-micron packed C(18) stationary phase (50 mm × 4.6 mm, 1.8 μm particle size) and a polar mobile phase of ACN:H(2)O:acetic acid (70:30:0.05) (v/v). The column temperature was maintained at (30±2°C), the injection volume was (8 μl), with a flow rate of (1.1 ml/min). Dermal microdialysis was used to collect (20 μl) samples from healthy, peri-lesional and lesional skin regions, in the forearms of a small cohort of subjects (n=6), and the ultra sensitive liquid chromatographic method allowed for nanomolar quantitation of histamine in 6.7 min. To date this represents one of the fastest reported separations of histamine using fluorescence detection with very high chromatographic efficiency (258,000/m) and peak symmetry of (0.88). Prior to sample analysis being performed method linearity, precision and limit of detection (LOD) were investigated. The results showed that intracutaneous histamine measured at 70 min after catheter

  15. 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.

  16. [Rapid quantification of total nitrogen and end-point determination of hide melting in manufacturing of donkey-hide gelatin].

    PubMed

    Han, Hai-Fan; Zhang, Lu; Zhang, Yan; Li, Wen-Long; Qu, Hai-Bin

    2014-03-01

    Hide melting presents itself as one of the most critical processes in the production of donkey-hide gelatin. Here a NIR-based method was established for the rapid analysis of in-process hide melting solutions as well as for end-point determination of this process. Near infrared (NIR) spectra of hide melting solutions were collected in transflective mode. With the contents of total nitrogen determined by the Kjeldahl method as reference values, partial least squares regression (PLSR) was employed to build calibration models between NIR spectra and total nitrogen. Model parameters including wavelength range and PLS factors were optimized to achieve best model performance. Based on the contents of total nitrogen predicted by calibration model, end point of hide melting was determined. The constructed PLS model gave a high correlation coefficient (R2) of 0.991 3 and a root mean square error of prediction (RMSEP) of 0.807 g x L(-1). With the predicted total nitrogen and predefined limit, decisions concerning the proper times of melting were made. This research demonstrated that NIR transflectance spectroscopy could be used to expeditiously determine the contents of total nitrogen which was subsequently chosen as the indictor for determining the end-point of hide melting. The proposed procedure may help avoid unnecessary raw material or energy consumption.

  17. Development of a rapid method for the quantification of cellulose in tobacco by (13)C CP/MAS NMR.

    PubMed

    Jiang, Jinhui; Hu, Yonghua; Tian, Zhenfeng; Chen, Kaibo; Ge, Shaolin; Xu, Yingbo; Tian, Dong; Yang, Jun

    2016-01-01

    A method was developed for rapid quantitative determination of cellulose in tobacco by utilizing (13)C cross polarization magic angle spinning NMR spectroscopy ((13)C CP/MAS NMR). Sample powder was loaded into NMR rotor, which was customized rotor containing a matched silicon tube as an intensity reference. (13)C CP/MAS NMR spectra of tobacco samples were processed with spectral deconvolution to obtain the area of the C-1 resonance at 105.5ppm and the internal standard at 0ppm. The ratio between the area of 105.5ppm and 0ppm of a set of standard cellulose samples was used to construct a calibration curve. The cellulose content of a tobacco sample was determined by comparison of the ratio between the area of 105.5ppm and 0ppm to the calibration curve. Results of this developed method showed good agreement with those obtained from chemical analysis. The proposed method has such advantages of accuracy, quickness and efficiency, and could be an alternative to chemical analyses of cellulose.

  18. Optimetric system facilitates colorimetric and fluorometric measurements

    NASA Technical Reports Server (NTRS)

    Haley, F. C.

    1968-01-01

    Compact, unitary optimetric systems uses a single device for colorimetric, fluorometric and spectral absorption measurements. The basic element of the unitary systems is a test cell containing filter elements with uniquely fabricated lenses.

  19. Colorimetric Coupled Enzyme Assay for Cystathionine β-Synthase.

    PubMed

    Rocchiccioli, Marco; Moschini, Roberta; Cappiello, Laura; Balestri, Francesco; Cappiello, Mario; Mura, Umberto; Del-Corso, Antonella

    2016-01-01

    A colorimetric coupled enzyme assay for the determination of cystathionine β-synthase activity is described. The method exploits cystathionine γ-lyase as an ancillary enzyme capable of transforming cystathionine, produced by cystathionine β-synthase, into cysteine. The cysteine is then spectrophotometrically detected at 560 nm, after its specific complexation with ninhydrin. This method was used to detect cystathionine β-synthase in crude extracts, and for the kinetic characterization of the enzyme partially purified from bovine kidney. A rapid two-step protocol is described for the partial purification of cystathionine γ-lyase from bovine kidney, aimed at a suitable and stable ancillary enzyme preparation. PMID:27506718

  20. 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

  1. 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

  2. Rapid HPLC Quantification Approach for Detection of Active Constituents in Modern Combinatorial Formula, San-Huang-Xie-Xin-Tang (SHXXT)

    PubMed Central

    Wu, Tung-Ying; Chang, Fang-Rong; Liou, Jing-Ru; Lo, I-Wen; Chung, Tang-Chia; Lee, Li-Yao; Chi, Chun-Chen; Du, Ying-Chi; Wong, Man-Hon; Juo, Suh-Hang Hank; Lee, Chun-Chen; Wu, Yang-Chang

    2016-01-01

    San-Huang-Xie-Xin-Tang (SHXXT), one of the most important traditional Chinese medicinal formulas, is comprised by three herbal medicines, the rhizome of Rheum officinale [or Rheum tanguticum (Polygonaceae) (Dahuang in Chinese)], the root of Scutellaria baicalensis (Labiatae) (Huangqin in Chinese), and the rhizome of Coptis chinensis (Ranunculaceae) (Huanglian in Chinese) in the ratios of 2:1:1 or 1:1:1. This study is aimed to quantitate and qualify of SHXXT, by a rapid, convenient, and effective HPLC-PDA approach associated with LC-MS technique. Of which method, nine chosen major bioactive components in SHXXT, including aloe-emodin (Ale), baicalin (Ba), berberine (Be), coptisine (Co), palmatine (Pa), resveratroloside (Res), rhein (Rh), sennoside A (Se-A), and wogonin (Wo), were evaluated within 30 min. The nine chemical markers were monitored in a high sensitivity with a low detection limit of 0.01−0.55 μg/mL and the correlation coefficient of the regression curve revealed a good linearity with R2 > 0.99. Moreover, the extraction solution system and the HPLC elution conditions were also optimized in the present study. This present developed protocol was then successfully applied to quantify nine chemical markers of 10 SHXXT products from eight Taiwanese TCM pharmaceutical companies. In quantitative results, Res was found as the major compound in SHXXT-1~5 and 8 with significantly higher amounts than those in other products, indicating the products SHXXT-1~5 and 8 may use R. tanguticum as the raw material, which possessed a higher concentration of the bioactive composition Res, instead of R. officinale. Simultaneously, Ale, Rh, and Wo were < 2% in these 10 products. Different chemical profiles of commercial products indicated that, probably, each product with the same named formula might be regarded as a sole medicine and need to be investigated individually. Importantly, it is never too much to emphasize the importance of quality control in TCM development. PMID

  3. Rapid and sensitive ultra-high-pressure liquid chromatography method for quantification of antichagasic benznidazole in plasma: application in a preclinical pharmacokinetic study.

    PubMed

    Davanço, Marcelo Gomes; de Campos, Michel Leandro; Peccinini, Rosângela Gonçalves

    2015-07-01

    Benznidazole (BNZ) and nifurtimox are the only drugs available for treating Chagas disease. In this work, we validated a bioanalytical method for the quantification of BNZ in plasma aimed at improving sensitivity and time of analysis compared with the assays already published. Furthermore, we demonstrated the application of the method in a preclinical pharmacokinetic study after administration of a single oral dose of BNZ in Wistar rats. A Waters® Acquity UHPLC system equipped with a UV-vis detector was employed. The method was established using an Acquity® UHPLC HSS SB C18 protected by an Acquity® UHPLC HSS SB C18 VanGuard guard column and detection at 324 nm. The mobile phase consisted of ultrapure water-acetonitrile (65:35), and elution was isocratic. The mobile phase flow rate was 0.55 mL/min, the volume of injection was 1 μL, and the run time was just 2 min. The samples were kept at 25°C until injection and the column at 45°C for the chromatographic separation. The sample preparation was performed by a rapid protein precipitation with acetonitrile. The linear concentration range was 0.15-20 µg/mL. The pharmacokinetic parameters of BNZ in rats were determined and the method was considered sensitive, fast and suitable for application in pharmacokinetic studies.

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

    PubMed

    Bidault, Adeline; 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 CECT4600(T) 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 10(1) 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

  5. 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

  6. Human nails metabolite analysis: A rapid and simple method for quantification of uric acid in human fingernail by high-performance liquid chromatography with UV-detection.

    PubMed

    Li, Xi-Ling; Li, Gao; Jiang, Ying-Zi; Kang, Dongzhou; Jin, Cheng Hua; Shi, Qing; Jin, Toufeng; Inoue, Koichi; Todoroki, Kenichiro; Toyo'oka, Toshimasa; Min, Jun Zhe

    2015-10-01

    A rapid and simple analytical method for the quantification of uric acid (UA) in human fingernails by high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection is described. UA was extracted from human fingernail samples at 90°C for 20min, then separated on an Inertsil ODS-2 column (250×4.6mm I.D., 5.0μm, GL Sciences) by isocratic elution using methanol: 74mM phosphate buffer (pH 2.2) 2:98 (v/v). An UV detector was used to monitor at 284nm. The results indicated that under optimized measurement conditions results were achieved within 8.0min, and a good linearity was achieved from the calibration curves (r(2)>0.9999) in the range of 1.0-10000ng; the limit of detection (S/N=3) was 2.0pg, the inter-day and intra-day assay precisions were all less than 0.46% and the mean recoveries (%) of the uric acid spiked in the human fingernail were 101.95%. The amounts of UA in the fingernails of healthy volunteers were determined.

  7. Simultaneous quantification of cilostazol and its primary metabolite 3,4-dehydrocilostazol in human plasma by rapid liquid chromatography/tandem mass spectrometry.

    PubMed

    Nirogi, Ramakrishna V S; Kandikere, Vishwottam N; Shukla, Manoj; Mudigonda, Koteshwara; Shrivasthava, Wishu; Datla, Praveen V; Yerramilli, Anjaneyulu

    2006-02-01

    A simple, rapid, sensitive and selective liquid chromatography/electrospray tandem mass spectrometry method was developed and validated for the simultaneous quantification of cilostazol and its primary metabolite 3,4-dehydrocilostazol in human plasma using mosapride as an internal standard. The method involves a simple one-step liquid-liquid extraction with a diethyl ether and dichloromethane mixture (7:3). The analytes were chromatographed using an isocratic mobile phase on a reversed-phase C18 column and analyzed by mass spectrometry in the multiple reaction monitoring mode using the respective [M+H]+ ions, m/z 370/288 for cilostazol, m/z 368/286 for 3,4-dehydrocilostazol and m/z 422/198 for the internal standard. The assay exhibited a linear dynamic range of 5-2,000 ng/mL for cilostazol and 5-400 ng/mL for 3,4-dehydrocilostazol in human plasma. The lower limit of quantitation was 5 ng/mL for both cilostazol and its metabolite. Acceptable precision and accuracy were obtained for concentrations over the standard curve ranges. A run time of 2.5 min for each sample made it possible to analyze more than 400 human plasma samples per day. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetics, bioavailability or bioequivalence studies.

  8. 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.

  9. 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

  10. 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.

  11. 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…

  12. 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...

  13. 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

  14. Rapid identification and quantification of methamphetamine and amphetamine in hair by gas chromatography/mass spectrometry coupled with micropulverized extraction, aqueous acetylation and microextraction by packed sorbent.

    PubMed

    Miyaguchi, Hajime; Iwata, Yuko T; Kanamori, Tatsuyuki; Tsujikawa, Kenji; Kuwayama, Kenji; Inoue, Hiroyuki

    2009-05-01

    We developed a rapid identification and quantification method for the toxicological analysis of methamphetamine and amphetamine in human hair by gas chromatography/mass spectrometry coupled with a novel combination of micropulverized extraction, aqueous acetylation and microextraction by packed sorbent (MEPS) named MiAMi-GC/MS. A washed hair sample (1-5 mg) was micropulverized for 5 min in a 2 mL plastic tube with 250 microL of water. An anion-exchange sorbent was added to adsorb anionic interferences. After removing the residue with a membrane-filter unit, sodium carbonate and acetic anhydride was admixed in turn. Acetylation was completed in approximately 20 min at room temperature. The acetylated analytes in the reaction liquid were concentrated to an octadecylsilica sorbent packed in the needle of a syringe by a CombiPAL autosampler. Elution was carried out with 50 microL of methanol, and the entire eluate injected into a gas chromatograph using a programmable temperature vaporizing (PTV) technique. The time required for sample preparation and GC/MS analysis was approximately 1 h from a washed hair sample, and an evaporation process was not required. Ranges for quantification were 0.20-50 (ng/mg) each for methamphetamine and amphetamine using 1 mg of hair. Accuracy and relative standard deviation (RSD) were evaluated intraday and interday at three concentrations, and the results were within the limit of a guidance issued by U.S. Food and Drug Administration. For identification, full-scan mass spectra of methamphetamine and amphetamine were obtained using 5 mg of fortified hair samples at 0.2 ng/mg. The extraction device of MEPS was durable for at least 300 extractions, whereas the liner of the gas chromatograph should be replaced after 20-30 times use. The carry over was estimated to be about 1-2%. This sample-preparation method coupled with GC/MS is fast and labor-saving in comparison with conventional methods.

  15. 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

  16. 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.

  17. [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.

  18. 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.

  19. 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.

  20. 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

  1. DNA-based methodologies for rapid detection, quantification, and species- or strain-level identification of respiratory pathogens (Mycobacteria and Pseudomonads) in metalworking fluids.

    PubMed

    Yadav, Jagjit S; Khan, Izhar U H; Fakhari, Farnaz; Soellner, Mathew B

    2003-11-01

    Mycobacteria and pseudomonads occurring in modern metalworking fluids (MWF) have been implicated in occupational health hazards as causal agents for hypersensitivity pneumonitis (HP) and other respiratory illnesses in machine workers exposed to these fluids and their aerosols. Unlike the conventional cultural and biochemical methods, which are often slow and ambiguous and detect only culturable cells, DNA-based methods offer a time-saving alternative for reliable detection and identification of both culturable and nonculturable bacteria in MWF and for selective quantification of individual genera of pathogens of interest in these fluids. This is the first report on DNA-based direct detection of mycobacteria and pseudomonads in MWF without culturing. Genus-specific PCR approach was successfully applied for screening of field MWF samples originating from different industrial users for detection of mycobacteria or pseudomonads including both culturable and nonculturable cells. PCR in combination with amplicon DNA sequencing led to the identification of Mycobacterium chelonae, Pseudomonas nitroreducens, and an undefined Pseudomonas species from these fluids. Genome fingerprinting by pulsed-field gel electrophoresis (PFGE) on Mycobacterium isolates further showed that the isolates represented three strains of M. chelonae although the possibility of one of the strains being clonal with M. immunogenum cannot be excluded. In parallel efforts, a quantitative competitive PCR method developed based on the Pseudomonas-specific PCR was applied to quantify total P. fluorescens cells in contaminated metalworking fluid and MWF aerosol without culturing. The DNA-based protocols developed in this study will allow rapid screening of field MWF samples for the presence of both culturable and nonculturable cells and thus facilitate effective fluid management and timely exposure assessment. PMID:14555451

  2. 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).

  3. Functionalized quantum dots with dopamine dithiocarbamate as the matrix for the quantification of efavirenz in human plasma and as affinity probes for rapid identification of microwave tryptic digested proteins in MALDI-TOF-MS.

    PubMed

    Kailasa, Suresh Kumar; Wu, Hui-Fen

    2012-06-01

    Functionalized quantum dots with dopamine dithiocarbamate (QDs-DDTC) were utilized for the first time as an efficient material for the quantification of efavirenz in human plasma of HIV infected patients and rapid identification of microwave tryptic digest proteins (cytochrome c, lysozyme and BSA) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The synthesized QDs-DDTC was characterized by using spectroscopic (UV-visible, FT-IR and (1)H NMR) and microscopic (SEM and TEM) techniques. Functionalized QDs-DDTC exhibited a high desorption/ionization efficiency for the rapid quantification of small molecules (efavirenz, tobramycin and aspartame) at low-mass region. QDs-DDTC has well ability to trap target species, and capable to transfer laser energy for efficient desorption/ionization of analytes with background-free detection. The use of QDs-DDTC as a matrix provided good linearity for the quantification of small molecules (R(2)=~0.9983), with good reproducibility (RSD<10%), in the analysis of efavirenz in the plasma of HIV infected patients by the standard addition method. We also demonstrated that the use of functionalized QDs-DDTC as affinity probes for the rapid identification of microwave tryptic digested proteins (cytochrome c, lysozyme and BSA) by MALDI-TOF-MS. QDs-DDTC-based MALDI-TOF-MS approach provides simplicity, rapidity, accuracy, and precision for the determination of efavirenz in human plasma of HIV infected patients and rapid identification of microwave tryptic digested proteins. This new material presents a marked advance in the development of matrix-free mass spectrometric methods for the rapid and precise quantitative determination of a variety of molecules. This article is part of a Special Issue entitled: Proteomics: The clinical link.

  4. 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.

  5. Multidimensional colorimetric sensor array for discrimination of proteins.

    PubMed

    Mao, Jinpeng; Lu, Yuexiang; Chang, Ning; Yang, Jiaoe; Zhang, Sichun; Liu, Yueying

    2016-12-15

    An extensible multidimensional colorimetric sensor array for the detection of protein is developed based on DNA functionalized gold nanoparticles (DNA-AuNPs) as receptors. In the presence of different proteins, the aggregation behavior of DNA-AuNPs was regulated by the high concentrations of salt and caused different color change; while DNA-AuNPs grew induced by the reduction of HAuCl4 and NH2OH as a reductant on the surface of nanoparticles exhibited different morphologies and color appearance for different proteins. The transducers based on AuNPs modified by specific and nonspecific DNA enables naked-eye discrimination of the target analytes. This extensible sensing platform with only two receptors could simultaneously discriminate ten native proteins and their thermally denatured conformations using hierarchical cluster analysis (HCA) at the concentration of 50nM with 100% accuracy. This opens up the possibility of the sensor array to investigate the different conformational changes of biomacromolecules, and it gives a new direction of developing multidimensional transduction principles based on plasmonic nanoparticle conjugates. Furthermore, the sensing system could discriminate proteins at the concentration of 500nM in the presence of 50% human urine, which indicated this sensor array has great potential ability in analyzing real biological fluids. In addition, the multidimensional colorimetric sensor array is suitable for analysis of target analytes in the resource-restricted regions because of rapid, simple, low cost, and in-field detection with the naked eye. PMID:27322936

  6. 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

  7. 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.

  8. Colorimetric Titration Experiment for the Undergraduate Laboratory.

    ERIC Educational Resources Information Center

    Lopez, Edwin; Vassos, Basil H.

    1984-01-01

    Describes a colorimetric titration instrument usable in the undergraduate laboratory that fulfills the objectives of ruggedness, freedom from ambient light interference, and low cost. Although accessories can be added (raising the price), the basic instrument is low priced and can be used manually with a simple voltmeter. (JN)

  9. 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…

  10. 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.

  11. Rapid and sensitive colorimetric determination of cobalt(II).

    PubMed

    Eldawy, M A; Tawfik, A S; Elshabouri, S R

    1976-05-01

    A highly selective and sensitive spectrophotometric determination of cobalt (II) was developed. 7-Nitroso-8-hydroxyquinoline-5-sulfonic acid sodium salt was used as the chromogenic reagent for color development. Although other metals form colored chelates with the ligand, it was possible to develop a selective method using McIlvaine's pH 8 citric acid-phosphate buffer. Under these conditions, iron(II), iron (III), copper (II), zinc (II), and manganese (II), minerals likely to be compounded with cobalt (II) in geriatric formulations, do not interfere with the precision of the method or the color development. Calcium (II) and magnesium (II) do not form colored chelates with the used ligand. Hormones, vitamins, and additives likely to be present along with the cobalt ion in pharmaceutical formulations do not interfere. The sensitivity is 0.37 mug of cobalt (II)/ml of sample solution.

  12. 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.

  13. 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.

  14. 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

  15. Highly selective colorimetric bacteria sensing based on protein-capped nanoparticles.

    PubMed

    Qiu, Suyan; Lin, Zhenyu; Zhou, Yaomin; Wang, Donggen; Yuan, Lijuan; Wei, Yihua; Dai, Tingcan; Luo, Linguang; Chen, Guonan

    2015-02-21

    A rapid and cost-effective colorimetric sensor has been developed for the detection of bacteria (Bacillus subtilis was selected as an example). The sensor was designed to rely on lysozyme-capped AuNPs with the advantages of effective amplification and high specificity. In the sensing system, lysozyme was able to bind strongly to Bacillus subtilis, which effectively induced a color change of the solution from light purple to purplish red. The lowest concentration of Bacillus subtilis detectable by the naked eye was 4.5 × 10(3) colony-forming units (CFU) mL(-1). Similar results were discernable from UV-Vis absorption measurements. A good specificity was observed through a statistical analysis method using the SPSS software (version 17.0). This simple colorimetric sensor may therefore be a rapid and specific method for a bacterial detection assay in complex samples. PMID:25503063

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

    PubMed

    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 (R (2)) > 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.

  17. 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

  18. 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.

  19. 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

  20. 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.

  1. 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

    Kanitkar, Yogendra H; Stedtfeld, Robert D; Steffan, Robert J; Hashsham, Syed A; Cupples, Alison M

    2016-01-08

    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.

  2. 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

  3. 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

    Kanitkar, Yogendra H; Stedtfeld, Robert D; Steffan, Robert J; Hashsham, Syed A; Cupples, Alison M

    2016-03-01

    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

  4. 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

  5. Visual detection of organophosphorus pesticides represented by mathamidophos using Au nanoparticles as colorimetric probe.

    PubMed

    Li, Hongkun; Guo, Jiajia; Ping, Hong; Liu, Lurui; Zhang, Minwei; Guan, Fengrui; Sun, Chunyan; Zhang, Qian

    2011-12-15

    With citrate-coated Au nanoparticles as colorimetric probe, a novel visual method for rapid assay of organophosphorus pesticides has been developed. The assay principle is based on catalytic hydrolysis of acetylthiocholine into thiocholine by acetylcholinesterase, which induces the aggregation of Au nanoparticles and the color change from claret-red to purple or even grey. The original plasmon absorption of Au nanoparticles at 522 nm decreases, and simultaneously, a new absorption band appears at 675 nm. The irreversible inhibition of organophosphorus pesticides on acetylcholinesterase prevents aggregation of Au nanoparticles. Under optimum conditions, the absorbance at 522 nm of Au nanoparticles is related linearly to the concentration of mathamidophos in the range of 0.02-1.42 μg/mL with a detection limit of 1.40 ng/mL. This colorimetric method has been successfully utilized to detect mathamidophos in vegetables with satisfactory results. The proposed colorimetric assay exhibits good reproducibility and accuracy, providing a simple and rapid method for the analysis of organophosphorus pesticides.

  6. 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

  7. 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.

  8. 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

  9. Colorimetric determination of phosphoric acid leakage for phosphoric acid-doped polybenzimidazole membrane fuel cell applications

    NASA Astrophysics Data System (ADS)

    Jeong, Yeon Hun; Jung, Ju Hae; Choi, Euiji; Han, Seungyoon; Begley, Alina Irene; Yoo, Sung Jong; Jang, Jong Hyun; Kim, Hyoung-Juhn; Nam, Suk Woo; Lee, Kwan-Young; Kim, Jin Young

    2015-12-01

    A simple and precise colorimetric method for analyzing phosphoric acid leakage in phosphoric acid-doped polybenzimidazole membrane fuel cells is described. The developed method is based on the colorimetric determination from a rapid formation of molybdenum blue color by the reduction reaction of molybdate ions in the presence of phosphoric acid in the acidic medium. The color is stable up to a few months and can be used for the sensitive and accurate detection of phosphoric acid electrolyte which is discharged from the fuel cell during operation. Tests with a wide concentration range of phosphate compounds showed that it permits determination of phosphoric acid up to nanogram quantities. The developed detection method assists monitoring the phosphoric acid contents and developing stable operation strategies of fuel cells.

  10. Colorimetric and ratiometric fluorescent detection of bisulfite by a new HBT-hemicyanine hybrid.

    PubMed

    Zhang, Haiyan; Huang, Zijun; Feng, Guoqiang

    2016-05-12

    A novel HBT-hemicyanine hybrid was prepared. This hybrid not only displays a large red-shifted (Δλ = 125 nm) emission compared to the well known ESIPT dye HBT, but also can be used as a new probe for rapid, colorimetric and ratiometric fluorescent detection of bisulfite with high selectivity and sensitivity in aqueous solution. The detection limit of this probe for HSO3(-) was calculated to be about 56 nM with a linear range of 0-25 μM. The potential application of this probe was exampled by detection of bisulfite in real food samples and living cells. Overall, this work not only provided a new ratiometric sensing platform, but also provided a new promising colorimetric and ratiometric fluorescent probe for bisulfite. PMID:27114225

  11. Quantification of Rapid Myosin Regulatory Light Chain Phosphorylation Using High-Throughput In-Cell Western Assays: Comparison to Western Immunoblots

    PubMed Central

    Aguilar, Hector N.; Zielnik, Barbara; Tracey, Curtis N.; Mitchell, Bryan F.

    2010-01-01

    Background Quantification of phospho-proteins (PPs) is crucial when studying cellular signaling pathways. Western immunoblotting (WB) is commonly used for the measurement of relative levels of signaling intermediates in experimental samples. However, WB is in general a labour-intensive and low-throughput technique. Because of variability in protein yield and phospho-signal preservation during protein harvesting, and potential loss of antigen during protein transfer, WB provides only semi-quantitative data. By comparison, the “in-cell western” (ICW) technique has high-throughput capacity and requires less extensive sample preparation. Thus, we compared the ICW technique to WB for measuring phosphorylated myosin regulatory light chain (PMLC20) in primary cultures of uterine myocytes to assess their relative specificity, sensitivity, precision, and quantification of biologically relevant responses. Methodology/Principal Findings ICWs are cell-based microplate assays for quantification of protein targets in their cellular context. ICWs utilize a two-channel infrared (IR) scanner (Odyssey®) to quantify signals arising from near-infrared (NIR) fluorophores conjugated to secondary antibodies. One channel is dedicated to measuring the protein of interest and the second is used for data normalization of the signal in each well of the microplate. Using uterine myocytes, we assessed oxytocin (OT)-stimulated MLC20 phosphorylation measured by ICW and WB, both using NIR fluorescence. ICW and WB data were comparable regarding signal linearity, signal specificity, and time course of phosphorylation response to OT. Conclusion/Significance ICW and WB yield comparable biological data. The advantages of ICW over WB are its high-throughput capacity, improved precision, and reduced sample preparation requirements. ICW might provide better sensitivity and precision with low-quantity samples or for protocols requiring large numbers of samples. These features make the ICW technique an

  12. A rapid and sensitive UPLC-MS/MS method for the simultaneous quantification of serum androsterone glucuronide, etiocholanolone glucuronide, and androstan-3α, 17β diol 17-glucuronide in postmenopausal women.

    PubMed

    Ke, Yuyong; Gonthier, Renaud; Isabelle, Maxim; Bertin, Jonathan; Simard, Jean-Nicolas; Dury, Alain Y; Labrie, Fernand

    2015-05-01

    Quantification of steroidal glucuronide conjugates by the indirect methods of immunoassay and GC-MS/MS may underestimate some conjugates since hydrolysis is needed in sample processing. In the present work, a sensitive and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the simultaneous direct quantification of androsterone glucuronide, etiocholanolone glucuronide, and androstan-3α, 17β diol 17-glucuronide in postmenopausal women's serum. The quantification limits are 0.1ng/mL for 3α-diol-17G and 4ng/mL for both ADT-G and Etio-G, respectively, with an extraction from 200μL serum while the total run time is less than 6min for all three glucuronides. In this method, solid phase extraction is used for sample preparation. The assay has been validated in compliance with EndoCeutics SOPs and FDA guidelines for bioanalytical method development and validation. The recovery of glucuronides in stripped serum is consistent with that in unstripped serum, where the average difference in stripped and unstripped is less than 10%. A linear regression model fits well the standard curves of all three compounds with R≥0.99 where the weighting factor is 1/X. Interday accuracy and CV for all levels of QCs are within the range of 15% in both stripped and unstripped serum while all calibration curves are within the range of 6% except for LLOQs, which are within the range of 9%. Other parameters have also been assessed such as selectivity, matrix, lipemic and hemolysis effects as well as stabilities in solution and matrix. Incurred sample reanalysis has been performed with a result of over 93% within 20% of the original values. This reliable, sensitive and fast method is ready for large-scale clinical sample assays. PMID:25701608

  13. Evaluating the feasibility of using insecticide quantification kits (IQK) for estimating cyanopyrethroid levels for indoor residual spraying in Vanuatu

    PubMed Central

    2014-01-01

    Background The quality of routine indoor residual spraying (IRS) operations is rarely assessed because of the limited choice of methods available for quantifying insecticide content in the field. This study, therefore, evaluated a user-friendly, rapid colorimetric assay for detecting insecticide content after routine IRS operations were conducted. Methods This study was conducted in Tafea Province, Vanuatu. Routine IRS was conducted with lambda cyhalothrin. Two methods were used to quantify the IRS activities: 1) pre-spray application of small felt pads and 2) post-spray removal of insecticide with adhesive. The insecticide content was quantified using a colorimetric assay (Insecticide Quantification Kit [IQK]), which involved exposing each sample to the test reagents for 15 mins. The concentration of insecticide was indicated by the depth of red colour. Results The IQK proved simple to perform in the field and results could be immediately interpreted by the programme staff. The insecticide content was successfully sampled by attaching felt pads to the house walls prior to spraying. The IRS operation was well conducted, with 83% of houses being sprayed at the target dose (20 – 30 mg AI/m2). The average reading across all houses was 24.4 ± 1.5 mg AI/m2. The results from the felt pads applied pre-spray were used as a base to compare methods for sampling insecticide from walls post-spray. The adhesive of Sellotape did not collect adequate samples. However, the adhesive of the felt pads provided accurate samples of the insecticide content on walls. Conclusion The IQK colorimetric assay proved to be a useful tool that was simple to use under realistic field conditions. The assay provided rapid information on IRS spray dynamics and spray team performance, facilitating timely decision making and reporting for programme managers. The IQK colorimetric assay will have direct applications for routine quality control in malaria control programmes globally and has the

  14. 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...

  15. 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

  16. Development and validation of an ultra-performance liquid chromatography quadrupole time of flight mass spectrometry method for rapid quantification of free amino acids in human urine.

    PubMed

    Joyce, Richard; Kuziene, Viktorija; Zou, Xin; Wang, Xueting; Pullen, Frank; Loo, Ruey Leng

    2016-01-01

    An ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-qTOF-MS) method using hydrophilic interaction liquid chromatography was developed and validated for simultaneous quantification of 18 free amino acids in urine with a total acquisition time including the column re-equilibration of less than 18 min per sample. This method involves simple sample preparation steps which consisted of 15 times dilution with acetonitrile to give a final composition of 25 % aqueous and 75 % acetonitrile without the need of any derivatization. The dynamic range for our calibration curve is approximately two orders of magnitude (120-fold from the lowest calibration curve point) with good linearity (r (2) ≥ 0.995 for all amino acids). Good separation of all amino acids as well as good intra- and inter-day accuracy (<15 %) and precision (<15 %) were observed using three quality control samples at a concentration of low, medium and high range of the calibration curve. The limits of detection (LOD) and lower limit of quantification of our method were ranging from approximately 1-300 nM and 0.01-0.5 µM, respectively. The stability of amino acids in the prepared urine samples was found to be stable for 72 h at 4 °C, after one freeze thaw cycle and for up to 4 weeks at -80 °C. We have applied this method to quantify the content of 18 free amino acids in 646 urine samples from a dietary intervention study. We were able to quantify all 18 free amino acids in these urine samples, if they were present at a level above the LOD. We found our method to be reproducible (accuracy and precision were typically <10 % for QCL, QCM and QCH) and the relatively high sample throughput nature of this method potentially makes it a suitable alternative for the analysis of urine samples in clinical setting.

  17. Comparison of macro-gravimetric and micro-colorimetric lipid determination methods.

    PubMed

    Inouye, Laura S; Lotufo, Guiherme R

    2006-10-15

    In order to validate a method for lipid analysis of small tissue samples, the standard macro-gravimetric method of Bligh-Dyer (1959) [E.G. Bligh, W.J. Dyer, Can. J. Biochem. Physiol. 37 (1959) 911] and a modification of the micro-colorimetric assay developed by Van Handel (1985) [E. Van Handel, J. Am. Mosq. Control Assoc. 1 (1985) 302] were compared. No significant differences were observed for wet tissues of two species of fish. However, limited analysis of wet tissue of the amphipod, Leptocheirusplumulosus, indicated that the Bligh-Dyer gravimetric method generated higher lipid values, most likely due to the inclusion of non-lipid materials. Additionally, significant differences between the methods were observed with dry tissues, with the micro-colorimetric method consistently reporting calculated lipid values greater than as reported by the gravimetric method. This was most likely due to poor extraction of dry tissue in the standard Bligh-Dyer method, as no significant differences were found when analyzing a single composite extract. The data presented supports the conclusion that the micro-colorimetric method described in this paper is accurate, rapid, and minimizes time and solvent use.

  18. Rapid detection and quantification of tyrosine decarboxylase gene (tdc) and its expression in gram-positive bacteria associated with fermented foods using PCR-based methods.

    PubMed

    Torriani, Sandra; Gatto, Veronica; Sembeni, Silvia; Tofalo, Rosanna; Suzzi, Giovanna; Belletti, Nicoletta; Gardini, Fausto; Bover-Cid, Sara

    2008-01-01

    In this study, PCR-based procedures were developed to detect the occurrence and quantify the expression of the tyrosine decarboxylase gene (tdc) in gram-positive bacteria associated with fermented foods. Consensus primers were used in conventional and reverse transcription PCR to analyze a collection of 87 pure cultures of lactic acid bacteria and staphylococci. All enterococci, Staphylococcus epidermidis, Lactobacillus brevis, Lactobacillus curvatus, and Lactobacillus fermentum strains and 1 of 10 Staphylococcus xylosus strains produced amplification products with the primers DEC5 and DEC3 in accordance with results of the screening plate method and with previously reported result obtained with high-performance liquid chromatography. No amplicons were obtained for tyramine-negative strains, confirming the high specificity of these new primers. A novel quantitative real-time PCR assay was successfully applied to quantify tdc and its transcript in pure cultures and in meat and meat products. This assay allowed estimation of the influence of different variables (pH, temperature, and NaCl concentration) on the tdc expression of the tyraminogenic strain Enterococcus faecalis EF37 after 72 h of growth in M17 medium. Data obtained suggest that stressful conditions could induce greater tyrosine decarboxylase activity. The culture-independent PCR procedures developed here may be used for reliable and fast detection and quantification of bacterial tyraminogenic activity without the limitations of conventional techniques.

  19. Rapid detection of immunity against bacteria in Asian honeybee and Western honeybee with quantification of royalisin in the hemolymphe by fast ELISA.

    PubMed

    Shen, Li-Rong; Dilireba, Shatar; Zhou, Wen-Xiu; Wang, Yi-Ran; Li, Mei-Lu; Zhai, Liang

    2014-09-24

    Royalisin from royal jelly (RJ) is a valuable peptide both for the prevention of honeybee diseases and for RJ preservation. ELISA for fast determination of royalisin content in hemolymphe (RCH) of honeybees with polyclonal antibody against recombinant royalisin from Asian honeybee was established. Assay on RCHs of health samples from Asian honeybee and Western honeybee showed the former (7.06 μg/mL) was significantly higher than that of the latter (5.64 μg/mL, p < 0.01). Moreover, relative to the non infection, the RCHs of Asian honeybees at 24 and 48 h post infection of Eschericha coli were higher than those of Western honeybees by 32.90% and 29.66%, respectively. Evidence revealed that Asian honeybee possesses higher innate immunity and immune response against bacteria in relation to the Western honeybee. The method will be a potential tool for detection of resistant levels to pathogens in honeybees and for quantification of royalisin in RJ products. PMID:25183454

  20. Rapid detection of immunity against bacteria in Asian honeybee and Western honeybee with quantification of royalisin in the hemolymphe by fast ELISA.

    PubMed

    Shen, Li-Rong; Dilireba, Shatar; Zhou, Wen-Xiu; Wang, Yi-Ran; Li, Mei-Lu; Zhai, Liang

    2014-09-24

    Royalisin from royal jelly (RJ) is a valuable peptide both for the prevention of honeybee diseases and for RJ preservation. ELISA for fast determination of royalisin content in hemolymphe (RCH) of honeybees with polyclonal antibody against recombinant royalisin from Asian honeybee was established. Assay on RCHs of health samples from Asian honeybee and Western honeybee showed the former (7.06 μg/mL) was significantly higher than that of the latter (5.64 μg/mL, p < 0.01). Moreover, relative to the non infection, the RCHs of Asian honeybees at 24 and 48 h post infection of Eschericha coli were higher than those of Western honeybees by 32.90% and 29.66%, respectively. Evidence revealed that Asian honeybee possesses higher innate immunity and immune response against bacteria in relation to the Western honeybee. The method will be a potential tool for detection of resistant levels to pathogens in honeybees and for quantification of royalisin in RJ products.

  1. Rapid detection and quantification of tyrosine decarboxylase gene (tdc) and its expression in gram-positive bacteria associated with fermented foods using PCR-based methods.

    PubMed

    Torriani, Sandra; Gatto, Veronica; Sembeni, Silvia; Tofalo, Rosanna; Suzzi, Giovanna; Belletti, Nicoletta; Gardini, Fausto; Bover-Cid, Sara

    2008-01-01

    In this study, PCR-based procedures were developed to detect the occurrence and quantify the expression of the tyrosine decarboxylase gene (tdc) in gram-positive bacteria associated with fermented foods. Consensus primers were used in conventional and reverse transcription PCR to analyze a collection of 87 pure cultures of lactic acid bacteria and staphylococci. All enterococci, Staphylococcus epidermidis, Lactobacillus brevis, Lactobacillus curvatus, and Lactobacillus fermentum strains and 1 of 10 Staphylococcus xylosus strains produced amplification products with the primers DEC5 and DEC3 in accordance with results of the screening plate method and with previously reported result obtained with high-performance liquid chromatography. No amplicons were obtained for tyramine-negative strains, confirming the high specificity of these new primers. A novel quantitative real-time PCR assay was successfully applied to quantify tdc and its transcript in pure cultures and in meat and meat products. This assay allowed estimation of the influence of different variables (pH, temperature, and NaCl concentration) on the tdc expression of the tyraminogenic strain Enterococcus faecalis EF37 after 72 h of growth in M17 medium. Data obtained suggest that stressful conditions could induce greater tyrosine decarboxylase activity. The culture-independent PCR procedures developed here may be used for reliable and fast detection and quantification of bacterial tyraminogenic activity without the limitations of conventional techniques. PMID:18236668

  2. 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

  3. 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.

  4. 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

  5. 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

  6. 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.

  7. Rapid reagentless quantification of alginate biosynthesis in Pseudomonas fluorescens bacteria mutants using FT-IR spectroscopy coupled to multivariate partial least squares regression.

    PubMed

    Correa, Elon; Sletta, Håvard; Ellis, David I; Hoel, Sunniva; Ertesvåg, Helga; Ellingsen, Trond E; Valla, Svein; Goodacre, Royston

    2012-07-01

    Alginate is an important medical and commercial product and currently is isolated from seaweeds. Certain microorganisms also produce alginate and these polymers have the potential to replace seaweed alginates in some applications, mainly because such production will allow much better and more reproducible control of critical qualitative polymer properties. The research conducted here presents the development of a new approach to this problem by analysing a transposon insertion mutant library constructed in an alginate-producing derivative of the Pseudomonas fluorescens strain SBW25. The procedure is based on the non-destructive and reagent-free method of Fourier transform infrared (FT-IR) spectroscopy which is used to generate a complex biochemical infrared fingerprint of the medium after bacterial growth. First, we investigate the potential differences caused by the growth media fructose and glycerol on the bacterial phenotype and alginate synthesis in 193 selected P. fluorescens mutants and show that clear phenotypic differences are observed in the infrared fingerprints. In order to quantify the level of the alginate we also report the construction and interpretation of multivariate partial least squares regression models which were able to quantify alginate levels successfully with typical normalized root-mean-square error in predictions of only approximately 14%. We have demonstrated that this high-throughput approach can be implemented in alginate screens and we believe that this FT-IR spectroscopic methodology, when combined with the most appropriate chemometrics, could easily be modified for the quantification of other valuable microbial products and play a valuable screening role for synthetic biology.

  8. Development of a rapid method for the sequential extraction and subsequent quantification of fatty acids and sugars from avocado mesocarp tissue.

    PubMed

    Meyer, Marjolaine D; Terry, Leon A

    2008-08-27

    Methods devised for oil extraction from avocado (Persea americana Mill.) mesocarp (e.g., Soxhlet) are usually lengthy and require operation at high temperature. Moreover, methods for extracting sugars from avocado tissue (e.g., 80% ethanol, v/v) do not allow for lipids to be easily measured from the same sample. This study describes a new simple method that enabled sequential extraction and subsequent quantification of both fatty acids and sugars from the same avocado mesocarp tissue sample. Freeze-dried mesocarp samples of avocado cv. Hass fruit of different ripening stages were extracted by homogenization with hexane and the oil extracts quantified for fatty acid composition by GC. The resulting filter residues were readily usable for sugar extraction with methanol (62.5%, v/v). For comparison, oil was also extracted using the standard Soxhlet technique and the resulting thimble residue extracted for sugars as before. An additional experiment was carried out whereby filter residues were also extracted using ethanol. Average oil yield using the Soxhlet technique was significantly (P < 0.05) higher than that obtained by homogenization with hexane, although the difference remained very slight, and fatty acid profiles of the oil extracts following both methods were very similar. Oil recovery improved with increasing ripeness of the fruit with minor differences observed in the fatty acid composition during postharvest ripening. After lipid removal, methanolic extraction was superior in recovering sucrose and perseitol as compared to 80% ethanol (v/v), whereas mannoheptulose recovery was not affected by solvent used. The method presented has the benefits of shorter extraction time, lower extraction temperature, and reduced amount of solvent and can be used for sequential extraction of fatty acids and sugars from the same sample.

  9. 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

  10. 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.

  11. Automated colorimetric screen for apyrase inhibitors.

    PubMed

    Windsor, J B; Thomas, C; Hurley, L; Roux, S J; Lloyd, A M

    2002-11-01

    Apyrases are enzymes that efficiently hydrolyze ATP and ADP and may operate both inside and outside the cell. Although apyrases are important to a variety of cellular mechanisms and uses in industry, there are no available apyrase-specific inhibitors. Colorimetric assays based on the Fiske-Subbarow method for measuring inorganic phosphate are able to detect the release of inorganic phosphate from ATP and other nucleotides. We found that this type of assay could be automated and used to screen for apyrase-inhibiting compounds by assaying for a reduction in released phosphate in the presence of potential inhibitors. The automation of this assay allowed for the successful screening of a commercially available compound library. Several low molecular weight compounds were identified that, when used at micromolar concentrations, effectively inhibited apyrase activity.

  12. Combined enzymatic and colorimetric method for determining the uronic acid and methylester content of pectin: Application to tomato products.

    PubMed

    Anthon, Gordon E; Barrett, Diane M

    2008-09-01

    A simple procedure for determining the galacturonic acid and methanol contents of soluble and insoluble pectins, relying on enzymatic pectin hydrolysis and colorimetric quantification, is described. Pectin samples are incubated with a commercial pectinase preparation, Viscozyme, then the galacturonic acid content of the hydrolyzed pectin is quantified colorimetrically using a modification of the Cu reduction procedure originally described by Avigad and Milner. This modification, substituting the commonly used Folin-Ciocalteau reagent for the arsenic containing Nelson reagent, gives a response that is linear, sensitive, and selective for uronic acids over neutral sugars. This method also avoids the use of concentrated acids needed for the commonly used m-phenylphenol method. Methanol, released by the action of the pectin methylesterase found in the Viscozyme, is quantified using alcohol oxidase and Purpald. This combined enzymatic and colorimetric procedure correctly determined the galacturonic acid and methanol content of purified, soluble citrus pectin. Application of the procedure to water insoluble pectins was evaluated with water insoluble material from apples and oranges. In both cases good agreement was obtained between this method and commonly used methods based on chemical pectin hydrolysis. Good agreement between these procedures was also found in the analysis of both soluble and insoluble pectins from several tomato products.

  13. 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

  14. 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.

  15. 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.

  16. Validation of an antibody-based biosensor for rapid quantification of 2,4,6-trinitrotoluene (TNT) contamination in ground water and river water.

    PubMed

    Bromage, Erin S; Vadas, George G; Harvey, Ellen; Unger, Michael A; Kaattari, Stephen L

    2007-10-15

    Nitroaromatics are common pollutants of soil and groundwater at military installations because of their manufacture, storage, and use at these sites. Long-term monitoring of these pollutants comprise a significant percentage of restoration costs. Further, remediation activities often have to be delayed, while the samples are processed via traditional chemical assessment protocols. Here we describe a rapid (<5 min), cost-effective, accurate method using a KinExA Inline Biosensor for monitoring of 2,4,6-trinitrotoluene (TNT) in field water samples. The biosensor, which is based on KinExA technology, accurately estimated the concentration of TNT in double-blind comparisons with similar accuracy to traditional high-performance liquid chromatography(HPLC). In the assessment of field samples, the biosensor accurately predicted the concentration of TNT over the range of 1-30,000 microg/L when compared to either HPLC or quantitative gas chromatography-mass spectrometry (GC-MS). Various pre-assessment techniques were explored to examine whether field samples could be assessed untreated, without the removal of particulates or the use of solvents. In most cases, the KinExA Inline Biosensor gave a uniform assessment of TNT concentration independent of pretreatment method. This indicates that this sensor possesses significant promise for rapid, on-site assessment of TNT pollution in environmental water samples.

  17. 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

  18. A novel strategy for rapid quantification of 20(S)-protopanaxatriol and 20(S)-protopanaxadiol saponins in Panax notoginseng P. ginseng and P. quinquefolium.

    PubMed

    Xu, Fa-Xiang; Yuan, Cen; Wan, Jian-Bo; Yan, Ru; Hu, Hao; Li, Shao-Ping; Zhang, Qing-Wen

    2015-01-01

    A novel strategy for the qualitative and quantitative determination of 20(S)-protopanaxatriol saponins (PTS) and 20(S)-protopanaxadiol saponins (PDS) in Panax notoginseng, Panax ginseng and Panax quinquefolium, based on the overlapping peaks of main components of PTS (calibrated by ginsenoside Rg1) and PDS (calibrated by ginsenoside Rb1), was proposed. The analysis was performed by using high-performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD). Under specific chromatographic conditions, all samples showed two overlapping peaks containing several main ginsenosides belonging to PTS and PDS, respectively. The overlapping peaks were also identified by using HPLC-MS. Based on the sum and ratio of PTS and PDS, 60 tested Panax samples were divided into three main clusters according to their species. The findings suggested that this strategy provides a simple and rapid approach to quantify PTS and PDS in Panax herbs.

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

    PubMed

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

    2012-11-01

    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.

  20. A G-quadruplex DNAzyme-based colorimetric method for facile detection of Alicyclobacillus acidoterrestris.

    PubMed

    Liu, Tao; Zhang, Xiao; Zhu, Wenxin; Liu, Wei; Zhang, Daohong; Wang, Jianlong

    2014-09-01

    The rapid and sensitive detection of Alicyclobacillus acidoterrestris (AA) has become very important due to the frequent occurrence of fruit juice spoilage by AA. In the present study, using guaiacol, both as the metabolic product of AA related to its concentration and as a green colorimetric substrate of G-quadruplex DNAzyme, a novel G-quadruplex DNAzyme-based colorimetric method for a rapid detection of AA has been developed for the first time. Under optimal conditions, AA has been successfully detected in the concentration range of 10(2)-10(5) cfu mL(-1) with a detection limit of 85 cfu mL(-1). The recoveries ranging from 71.8% to 115.7% with relative standard deviation from 1.2% to 6.6% in spiked apple and orange juice samples were obtained. Results demonstrate that the sensitivity and precision of the developed method is comparable with most other analytical methods and is prominently rapid than them. We believe that the work provides a novel and effective approach and is beneficial for monitoring and reducing the risk of AA contaminations during the process of fruit juice production.

  1. 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.

  2. 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

  3. 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

  4. A simple, rapid and sensitive UFLC-MS/MS method for the quantification of oral contraceptive norgestrel in human plasma and its pharmacokinetic applications.

    PubMed

    Batta, N; Pigili, R K; Pallapothu, L M K; Yejella, R P

    2014-09-01

    A simple, rapid and sensitive ultra flow liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) assay for the determination of norgestrel in human plasma was developed using levonorgestrel D6 as an internal standard (IS). Norgestrel and IS were extracted from human plasma via liquid-liquid extraction. Chromatographic separation was achieved on a Zorbax XDB-Phenyl column under isocratic conditions. Detection was done by tandem mass spectrometry, operating in positive ion mode. The protonated precursor to product ion transitions monitored for norgestrel and IS were at m/z 313.30→245.40 and 319.00→251.30 respectively. The method was fully validated as per current regulatory guidelines. Anticoagulant counter ion effect was also assessed with K2EDTA and K3EDTA. The method was validated with a linearity range of 304.356-50 807.337 pg/mL having run time of 2.0 min per sample. The method has shown tremendous reproducibility with intra- and inter-day precision (%CV) less than 11.0% and intra- and inter-day accuracy less than 9.0% of nominal values. The validated method was applied to a pharmacokinetic study in human plasma samples generated after administrating a single oral dose of 0.3 mg norgestrel tablets to healthy female volunteers and has proved to be highly reliable for the analysis of clinical samples. PMID:24310360

  5. 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

  6. 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.

  7. 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

  8. Avoiding artifacts in the infant rat model for bacterial meningitis: use of Sangur test strips for the rapid quantification of blood contamination in cerebrospinal fluid.

    PubMed

    Vogel, U; Steinmetz, I; Frosch, M

    1996-05-01

    The infant rat model is widely used to study the pathogenesis of meningitis caused by a variety of gram-positive and gram-negative bacteria. However, the interpretation of published results concerning meningitis is difficult in many records because the fact that blood contamination of the cerebrospinal fluid (CSF) cannot be avoided during the traumatic puncture procedure has not been taken into consideration. Since bacterial invasion of the central nervous system develops following bacteremia in this model, blood contamination of the CSF leads to a falsification of the CSF bacterial counts. Here we present an evaluation of a rapid and quantitative test for CSF blood contamination using Sangur test strips. The procedure requires minimal amounts of CSF and allows direct calculation of the CSF bacterial load due to blood contamination and, thus, provides refined criteria for the presence of bacterial meningitis in the infant rat model. It is superior to the detection of erythrocytes using a hemocytometer since it is less time consuming. Furthermore, we demonstrate the value of this method for the experimental infection of rats with Neisseria meningitis.

  9. 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

  10. 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

  11. 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.

  12. 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.

  13. 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

  14. 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.

  15. 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

  16. 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

  17. Quantification of viral proteins of the avian H7 subtype of influenza virus: an isotope dilution mass spectrometry method applicable for producing more rapid vaccines in the case of an influenza pandemic.

    PubMed

    Santana, Wanda I; Williams, Tracie L; Winne, Emily K; Pirkle, James L; Barr, John R

    2014-05-01

    Vaccination is the most effective means to prevent influenza and its serious complications. Influenza viral strains undergo rapid mutations of the surface proteins hemagglutinin (HA) and neuraminidase (NA) requiring vaccines to be frequently updated to include current circulating strains. It is nearly impossible to predict which strains will be circulating in the next influenza season. It is, therefore, imperative that the process of producing a vaccine be streamlined and as swift as possible. We have developed an isotope dilution mass spectrometry (IDMS) method to quantify HA and NA in H7N7, H7N2, and H7N9 influenza. The IDMS method involves enzymatic digestion of viral proteins and the specific detection of evolutionarily conserved target peptides. The four target peptides that were initially chosen for analysis of the HA protein of H7N2 and H7N7 subtypes were conserved and available for analysis of the H7N9 subtype that circulated in China in the spring of 2013. Thus, rapid response to the potential pandemic was realized. Quantification of a protein is performed by employing multiple peptides to ensure that the enzymatic digestion of the protein is efficient in the region of the target peptides, verify the accuracy of the measurement, and provide flexibility in the case of amino acid changes among newly emerging strains. The IDMS method is an accurate, sensitive, and selective method to quantify the amount of HA and NA antigens in primary liquid standards, crude allantoic fluid, purified virus samples, and final vaccine presentations.

  18. 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

  19. 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.

  20. Selective spectrophotometric determination of TNT using a dicyclohexylamine-based colorimetric sensor.

    PubMed

    Erçağ, Erol; Uzer, Ayşem; Apak, Reşat

    2009-05-15

    Because of the extremely heterogeneous distribution of explosives in contaminated soils, on-site colorimetric methods are efficient tools to assess the nature and extent of contamination. To meet the need for rapid and low-cost chemical sensing of explosive traces or residues in soil and post-blast debris, a colorimetric absorption-based sensor for trinitrotoluene (TNT) determination has been developed. The charge-transfer (CT) reagent (dicyclohexylamine, DCHA) is entrapped in a polyvinylchloride (PVC) polymer matrix plasticised with dioctylphtalate (DOP), and moulded into a transparent sensor membrane sliced into test strips capable of sensing TNT showing an absorption maximum at 530 nm when placed in a 1-mm spectrophotometer cell. The sensor gave a linear absorption response to 5-50 mg L(-1) TNT solutions in 30% aqueous acetone with limit of detection (LOD): 3 mg L(-1). The sensor is only affected by tetryl, but not by RDX, pentaerythritoltetranitrate (PETN), dinitrotoluene (DNT), and picric acid. The proposed method was statistically validated for TNT assay against high performance liquid chromatography (HPLC) using a standard sample of Comp B. The developed sensor was relatively resistant to air and water, was of low-cost and high specificity, gave a rapid and reproducible response, and was suitable for field use of TNT determination in both dry and humid soil and groundwater with a portable colorimeter.

  1. 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

  2. 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.

  3. 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

  4. 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

  5. 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.

  6. 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

  7. 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

  8. 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.

  9. 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.

  10. A colorimetric sandwich-type assay for sensitive thrombin detection based on enzyme-linked aptamer assay.

    PubMed

    Park, Jun Hee; Cho, Yea Seul; Kang, Sungmuk; Lee, Eun Jeong; Lee, Gwan-Ho; Hah, Sang Soo

    2014-10-01

    A colorimetric sandwich-type assay based on enzyme-linked aptamer assay has been developed for the fast and sensitive detection of as low as 25 fM of thrombin with high linearity. Aptamer-immobilized glass was used to capture the target analyte, whereas a second aptamer, functionalized with horseradish peroxidase (HRP), was employed for the conventional 3,5,3',5'-tetramethylbenzidine (TMB)-based colorimetric detection. Without the troublesome antibody requirement of the conventional enzyme-linked immunosorbent assay (ELISA), as low as 25 fM of thrombin could be rapidly and reproducibly detected. This assay has superior, or at least equal, recovery and accuracy to that of conventional antibody-based ELISA.

  11. A BODIPY derivative as a colorimetric, near-infrared and turn-on chemosensor for Cu2+.

    PubMed

    Yin, Shouchun; Yuan, Wenli; Huang, Jinlong; Xie, Danbo; Liu, Bin; Jiang, Kezhi; Qiu, Huayu

    2012-10-01

    A new colorimetric and near-infrared (NIR) fluorescent chemosensor (1) for Cu(2+) based on BODIPY has been synthesized and investigated in this work. 1 Displays a high selectivity for Cu(2+) with about 250-fold enhancement in fluorescence emission intensity and micromolar sensitivity (K(d)=2.8±0.3 μM) in comparison with alkali and alkaline earth metal ions (Na(+), K(+), Mg(2+), Ca(2+)) and other metal ions (Ba(2+), Zn(2+), Cd(2+), Fe(2+), Pb(2+), Ni(2+), Co(2+), Ag(+)) upon excitation at 635 nm in CH(3)CN. Meanwhile, the distinct color changes and rapid switch-on fluorescence also provide 1 as colorimetric senor for Cu(2+). PMID:22658999

  12. ``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

  13. 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.

  14. 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.

  15. EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans

    PubMed Central

    Sjögren, Jonathan; Cosgrave, Eoin F J; Allhorn, Maria; Nordgren, Maria; Björk, Stephan; Olsson, Fredrik; Fredriksson, Sarah; Collin, Mattias

    2015-01-01

    Enzymes that affect glycoproteins of the human immune system, and thereby modulate defense responses, are abundant among bacterial pathogens. Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. By analyzing the glycans hydrolyzed by EndoS and EndoS2 from the antibodies using matrix-assisted laser desorption ionization time of flight, we found that both the enzymes cleaved complex glycans and that EndoS2 hydrolyzed hybrid and oligomannose structures to a greater extent compared with EndoS. A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. We conclude that EndoS and EndoS2 hydrolyze different glycoforms from the Fc-glycosylation site on therapeutic mAbs and that this can be used for rapid quantification of high mannose content. PMID:26156869

  16. EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans.

    PubMed

    Sjögren, Jonathan; Cosgrave, Eoin F J; Allhorn, Maria; Nordgren, Maria; Björk, Stephan; Olsson, Fredrik; Fredriksson, Sarah; Collin, Mattias

    2015-10-01

    Enzymes that affect glycoproteins of the human immune system, and thereby modulate defense responses, are abundant among bacterial pathogens. Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. By analyzing the glycans hydrolyzed by EndoS and EndoS2 from the antibodies using matrix-assisted laser desorption ionization time of flight, we found that both the enzymes cleaved complex glycans and that EndoS2 hydrolyzed hybrid and oligomannose structures to a greater extent compared with EndoS. A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. We conclude that EndoS and EndoS2 hydrolyze different glycoforms from the Fc-glycosylation site on therapeutic mAbs and that this can be used for rapid quantification of high mannose content.

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. A colorimetric detection method of pesticide acetamiprid by fine-tuning aptamer length.

    PubMed

    Tian, Yu; Wang, Yuan; Sheng, Zhi; Li, Tingting; Li, Xu

    2016-11-15

    This work investigates the effect of shortening aptamer sequences on the colorimetric detection of acetamiprid using aptamer-wrapped gold nanoparticles (AuNPs). Truncated 37-mer and 25-mer aptamers were generated by deleting excess flanking nucleotides from parental 49-mer acetamiprid-target aptamer. In comparing the responses of the three sequences, truncated aptamers did not improve the ability to discriminate against other tested pesticides. However, comparison between 49-mer and other shorter aptamers showed that shortening aptamer sequences through removing excess flanking nucleotides outsides of binding region improved colorimetric sensitivity for acetamiprid by 3.3 fold. Due to excess bases, the target-bound aptamer might still adhere to AuNPs, resulting in incomplete dissociation of aptamer from AuNPs and therefore the suppression of aggregation responses. This work provides further insight to the effects of aptamer structure on detection of the target, as well as a method by fine-tuning aptamer length for rapid detection of pesticide residues in environments or food.

  2. 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.

  3. 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.

  4. A colorimetric assay for determination of cell viability in algal cultures.

    PubMed

    Capasso, Juan M; Cossío, Belén R; Berl, Tomás; Rivard, Christopher J; Jiménez, Carlos

    2003-07-01

    In this work, we propose the determination of cell viability in algal cultures by using a colorimetric assay widely used for estimation of cell proliferation in animal cell cultures. The method is based on in vivo reduction by metabolically active cells of a tetrazolium compound (MTS=3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenil)-2H-tetrazolium, inner salt) to a colored formazan, with maximal absorbance at 490 nm, that is released to the culture medium. For this purpose, we have tested two microalgae with high commercial value (Dunaliella and Spirulina) and two seaweeds with different morphology (Ulva and Gracilaria). Color development in this assay is directly proportional to the number of viable cells, to the incubation time in the presence of the assay solution, and to the incubation temperature. A direct significant correlation was found between algal photosynthesis rate and color development in all species used through this work. Moreover, the intensity of absorbance at 490 nm was significantly lower in stressed cells (e.g. in nutrient-limited cultures, in the presence of toxic substances, and in osmotically-stressed cultures). We conclude that cell viability of algal cultures can be rapidly and easily estimated through colorimetric determination of the reduction of MTS to formazan.

  5. 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.

  6. A colorimetric detection method of pesticide acetamiprid by fine-tuning aptamer length.

    PubMed

    Tian, Yu; Wang, Yuan; Sheng, Zhi; Li, Tingting; Li, Xu

    2016-11-15

    This work investigates the effect of shortening aptamer sequences on the colorimetric detection of acetamiprid using aptamer-wrapped gold nanoparticles (AuNPs). Truncated 37-mer and 25-mer aptamers were generated by deleting excess flanking nucleotides from parental 49-mer acetamiprid-target aptamer. In comparing the responses of the three sequences, truncated aptamers did not improve the ability to discriminate against other tested pesticides. However, comparison between 49-mer and other shorter aptamers showed that shortening aptamer sequences through removing excess flanking nucleotides outsides of binding region improved colorimetric sensitivity for acetamiprid by 3.3 fold. Due to excess bases, the target-bound aptamer might still adhere to AuNPs, resulting in incomplete dissociation of aptamer from AuNPs and therefore the suppression of aggregation responses. This work provides further insight to the effects of aptamer structure on detection of the target, as well as a method by fine-tuning aptamer length for rapid detection of pesticide residues in environments or food. PMID:27612649

  7. 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

  8. 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.

  9. Dual-responsive immunosensor that combines colorimetric recognition and electrochemical response for ultrasensitive detection of cancer biomarkers.

    PubMed

    Hong, Wooyoung; Lee, Sooyeon; Cho, Youngnam

    2016-12-15

    We developed a nanoroughened, biotin-doped polypyrrole immunosensor for the detection of tumor markers through dual-signal (electrochemical and colorimetric) channels, electrochemical and colorimetric, that demonstrates remarkable analytical performance. A rapid, one-step electric field-mediated method was employed to fabricate the immunosensor with nanoscale roughness by simply modulating the applied electrical potential. We demonstrated the successful detection of three tumor markers (CA125, CEA, and PSA) via the double enzymatic signal amplifications in the presence of a target antigen, ultimately leading to desired diagnostic accuracy and reliability. The addition of multiple horseradish peroxidase (HRP)- and antibody-labeled nanoparticles greatly amplified the signal and simplified the measurement of cancer biomarker proteins by sequentially magnifying electrochemical and colorimetric signals in a single platform. The two parallel assays performed using the proposed immunosensor have yielded highly consistent and reproducible results. Additionally, for the analysis of plasma samples in a clinical setting, the values obtained with our immunosensor were validated by correlating the results with those of a standard radioimmunoassay (RIA), which obtained very similar clinically valid responses. PMID:27497199

  10. 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…

  11. 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

  12. 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…

  13. A NaBH₄ Coupled Ninhydrin-Based Assay for the Quantification of Protein/Enzymes During the Enzymatic Hydrolysis of Pretreated Lignocellulosic Biomass.

    PubMed

    Mok, Yiu Ki; Arantes, Valdeir; Saddler, Jack N

    2015-07-01

    Accurate protein quantification is necessary in many of the steps during the enzymatic hydrolysis of pretreated lignocellulosic biomass, from the fundamental determination of enzyme kinetics to techno-economic assessments, such as the use of enzyme recycling strategies, evaluation of enzyme costs, and the optimization of various process steps. In the work described here, a modified, more rapid ninhydrin-based protein quantification assay was developed to better quantify enzyme levels in the presence of lignocellulosic biomass derived compounds. The addition of sodium borohydride followed by acid hydrolysis at 130 °C greatly reduced interference from monosaccharides and oligosaccharides and decreased the assay time 6-fold. The modified ninhydrin assay was shown to be more accurate as compared to various traditional colorimetric protein assays when commercial cellulase enzyme mixtures were quantified under typical pretreated lignocellulosic biomass enzymatic hydrolysis conditions. The relatively short assay time and microplate-reading capability of the modified assay indicated that the method could likely be used for high-throughput protein determination. PMID:25987134

  14. 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.

  15. Colorimetric Detection of Plasmodium vivax in Urine Using MSP10 Oligonucleotides and Gold Nanoparticles

    PubMed Central

    Alnasser, Yossef; Ferradas, Cusi; Clark, Taryn; Calderon, Maritza; Gurbillon, Alejandro; Gamboa, Dionicia; McKakpo, Uri S.; Quakyi, Isabella A.; Bosompem, Kwabena M.; Sullivan, David J.; Vinetz, Joseph M.; Gilman, Robert H.

    2016-01-01

    Plasmodium vivax is the most prevalent cause of human malaria in the world and can lead to severe disease with high potential for relapse. Its genetic and geographic diversities make it challenging to control. P. vivax is understudied and to achieve control of malaria in endemic areas, a rapid, accurate, and simple diagnostic tool is necessary. In this pilot study, we found that a colorimetric system using AuNPs and MSP10 DNA detection in urine can provide fast, easy, and inexpensive identification of P. vivax. The test exhibited promising sensitivity (84%), high specificity (97%), and only mild cross-reactivity with P. falciparum (21%). It is simple to use, with a visible color change that negates the need for a spectrometer, making it suitable for use in austere conditions. Using urine eliminates the need for finger-prick, increasing both the safety profile and patient acceptance of this model. PMID:27706158

  16. 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

  17. 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

  18. 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

  19. Development of a universal colorimetric indicator for G-quadruplex structures by the fusion of thiazole orange and isaindigotone skeleton.

    PubMed

    Yan, Jin-Wu; Ye, Wen-Jie; Chen, Shuo-Bin; Wu, Wei-Bin; Hou, Jin-Qiang; Ou, Tian-Miao; Tan, Jia-Heng; Li, Ding; Gu, Lian-Quan; Huang, Zhi-Shu

    2012-08-01

    The rapid and convenient method for identification of all kinds of G-quadruplex is highly desirable. In the present study, a novel colorimetric indicator for a vast variety of G-quadruplex was designed and synthesized on the basis of thiazole orange and isaindigotone skeleton. Its distinct color change enables label-free visual detection of G-quadruplexes, which is due to the disassembly of dye H-aggregates to monomers. This specific detection of G-quadruplex arises from its end-stacking interaction with G-quartet. On the basis of this universal indicator, a facile approach for large-scale identification of G-quadruplex was developed.

  20. An Amidochlorin-Based Colorimetric Fluorescent Probe for Selective Cu(2+) Detection.

    PubMed

    Li, Wenting; Zhu, Guohua; Li, Jinghua; Wang, Zhiqiang; Jin, Yingxue

    2016-01-01

    The design and synthesis of selective and sensitive chemosensors for the quantification of environmentally and biologically important ionic species has attracted widespread attention. Amidochlorin p6 (ACP); an effective colorimetric and fluorescent probe for copper ions (Cu(2+)) in aqueous solution derived from methyl pheophorbide-a (MPa) was designed and synthesized. A remarkable color change from pale yellow to blue was easily observed by the naked eye upon addition of Cu(2+); and a fluorescence quenching was also determined. The research of fluorescent quenching of ACP-Cu(2+) complexation showed the detection limit was 7.5 × 10(-8) mol/L; which suggested that ACP can act as a high sensitive probe for Cu(2+) and can be used to quantitatively detect low levels of Cu(2+) in aqueous solution. In aqueous solution the probe exhibits excellent selectivity and sensitivity toward Cu(2+) ions over other metal ions (M = Zn(2+); Ni(2+); Ba(2+); Ag⁺; Co(2+); Na⁺; K⁺; Mg(2+); Cd(2+); Pb(2+); Mn(2+); Fe(3+); and Ca(2+)). The obvious change from pale yellow to blue upon the addition of Cu(2+) could make it a suitable "naked eye" indicator for Cu(2+). PMID:26797591

  1. 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

  2. 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.

  3. 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).

  4. 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

  5. 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.

  6. Ionic Liquid-Based Fluorescein Colorimetric pH Nanosensors

    PubMed Central

    Das, Susmita; Magut, Paul K. S.; de Rooy, Sergio L.; Hasan, Farhana; Warner, Isiah M.

    2014-01-01

    A novel pH sensitive, colorimetric ionic liquid nanosensor based on phosphonium salts of fluorescein is reported. Herein, fluorescein salts of various stoichiometries were synthesized by use of a trihexyltetradecylphosphonium cation [TTP]+ in combination with dianionic [FL]2− and monoanionic [FL]− fluorescein. Nanomaterials derived from these two compounds yielded contrasting colorimetric responses in neutral and acidic environments. Variations in fluorescence spectra as a function of pH were also observed. Examination of TEM and DLS data revealed significant expansion in the diameter of [TTP]2[FL] nanodroplets in acidic environments of variable pHs. A similar trend was also observed for [TTP][FL] nanoparticles. The pH dependent colorimetric and other optical properties of these nanomaterials are attributed to alterations in molecular orientations and stacking as suggested by measuring the absorption, fluorescence, and zeta potential. Since the pH is an important indicator for many diseases, including cancer, these nanosensors are considered to be potential candidates for biomedical applications. PMID:25264488

  7. 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.

  8. 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.

  9. 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

  10. Colorimetric determination of pH

    SciTech Connect

    Baumann, E.W.; Buchanan, B.R.

    1991-12-31

    There is a need for a simple, rapid, reliable means for determining pH values of concentrated, high salt solutions without reliance on human eye and ambient light. The method comprises the steps of preparing a set of reference solutions, measuring the light absorption by each reference solution, adding indicator dye to each reference solution, measuring the light absorption by each such reference mixture, comparing the two solutions to determine the dye color at each pH, normalizing the spectra of mixture to the isosbestic point, and matching the color of the pH of the solution to one of the colors of the pHs in the reference solution set. In this way, the pH can be determined to within 0.1 pH unit, a far more precise method than using the human eye.

  11. Sensitive and rapid quantification of the cannabinoid receptor agonist naphthalen-1-yl-(1-pentylindol-3-yl)methanone (JWH-018) in human serum by liquid chromatography-tandem mass spectrometry.

    PubMed

    Teske, Jörg; Weller, Jens-Peter; Fieguth, Armin; Rothämel, Thomas; Schulz, Yvonne; Tröger, Hans Dieter

    2010-10-01

    The current paper describes a validated method for the detection and quantification of naphthalen-1-yl-(1-pentylindol-3-yl)methanone (JWH-018), an ingredient of a herbal mixture called "Spice", by means of HPLC-ESI-MS-MS in serum. Lower limit of detection and lower limit of quantification were 0.07 and 0.21 ng/ml, respectively. In 2 subjects who consumed ca. 50 μg/kg of JWH-018 by smoking, the active ingredient was detected by means of the described method. Thereby, the serum concentrations reached values of approx. 10 ng/ml and dropped within 3 h very fast (<10% of the measured maximum concentrations).

  12. Colorimetric determination of zirconium in antiperspirant aerosols.

    PubMed

    Beavin, P

    1976-07-01

    A rapid direct dilution procedure for the estimation of soluble zirconium and a fusion procedure for the determination of total zirconium (soluble and insoluble forms) in cream base concentrates prepared from antiperspirant aerosols are described. The direct dilution procedure involves extraction of soluble zirconium with HCl (55 + 45). The filtered extract is reacted with alizarin red S to form a stable colored complex which is measured spectrophotometrically. The fusion procedure involves ashing the aerosol concentrate followed by fusion of the ash with potassium pyrosulfate to form an acid-soluble melt. Zirconium is precipitated from solution as the hydroxide and washed to eliminate interfering ions, particularly sulfate. After redissolving in HCl (55 + 45) and reacting with alizarin red S, total zirconium is measured. Zirconyl chloride octahydrate, assayed gravimetrically by hydroxide precipitation and conversion to the oxide, is used as the zirconium reference standard. Concentration range of zirconium measured was 200-500 mug/100 ml. Recoveries of standard zirconium added to commercial aerosols labeled to contain aluminum and zirconyl hydroxychlorides ranged from 97 to 101% by the fusion procedure. Analysis of these aerosols by direct dilution gave generally slightly lower results than by fusion. It is recommended that the procedures be collaboratively studied after further testing of their general applicability to a variety of drugs and cosmetics. PMID:939750

  13. 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

  14. New function of exonuclease and highly sensitive label-free colorimetric DNA detection.

    PubMed

    Li, Hongbo; Wang, Suqin; Wu, Zaisheng; Xu, Jianguo; Shen, Guoli; Yu, Ruqin

    2016-03-15

    Enzymatic manipulation and modulation of nucleic acids are a central part of cellular function, protection, and reproduction, while rapid and accurate detection of ultralow amount of nucleic acids remains a major challenge in molecular biology research and clinic diagnosis of genetic diseases. Herein, we reported that exonuclease III can degrade the G-quadruplex structure, indicating the new exonuclease's function. Basing on the function of exonuclease III, a novel G-quadruplex-hemin DNAzyme-based colorimetric detection of tumor suppressor gene p53 was successfully developed. Although only one oligonucleotide probe was involved, the sensing strategy could suppress the optical background and achieve an efficient G-quadruplex-hemin DNAzyme-based signal amplification. Specifically, a label-free functional nucleic acid probe (called THzyme probe) was designed via introducing target DNA probe-contained hairpin structure into G-quadruplex DNAzyme. Even if this probe can fold into G-quadruplex structure in the presence of hemin very different from the double-stranded DNA, it is easily degraded by exonuclease III. Thus, no change in UV-vis absorption intensity is detected in the absence of target DNA. However, the hybridization of target DNA can protect the integrity and catalytic activity of THzyme probe, producing the DNAzyme-amplified colorimetric signal. As a result, the p53 gene was able to be detected down to 1.0 pM (final concentration in the signal-generating solution: 50.0 fM) and mismatched target DNAs were easily distinguished. It is expected that this simple sensing methodology for DNA detection can find its utility in point-of-care applications. PMID:26519729

  15. 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

  16. L-cysteine protected copper nanoparticles as colorimetric sensor for mercuric ions.

    PubMed

    Soomro, Razium A; Nafady, Ayman; Sirajuddin; Memon, Najma; Sherazi, Tufail H; Kalwar, Nazar H

    2014-12-01

    This report demonstrates a novel, simple and efficient protocol for the synthesis of copper nanoparticles in aqueous solution using L-cysteine as capping or protecting agent. UV-visible (UV-vis) spectroscopy was employed to monitor the LSPR band of L-cysteine functionalized copper nanoparticles (Cyst-Cu NPs) based on optimizing various reaction parameters. Fourier Transform Infrared (FTIR) spectroscopy provided information about the surface interaction between L-cysteine and Cu NPs. Transmission Electron Microscopy (TEM) confirmed the formation of fine spherical, uniformly distributed Cyst-Cu NPs with average size of 34 ± 2.1 nm. X-ray diffractometry (XRD) illustrated the formation of pure metallic phase crystalline Cyst-Cu NPs. As prepared Cyst-Cu NPs were tested as colorimetric sensor for determining mercuric (Hg(2+)) ions in an aqueous system. Cyst-Cu NPs demonstrated very sensitive and selective colorimetric detection of Hg(2+) ions in the range of 0.5 × 10(-6)-3.5 × 10(-6) mol L(-1) based on decrease in LSPR intensity as monitored by a UV-vis spectrophotometer. The developed sensor is simple, economic compared to those based on precious metal nanoparticles and sensitive to detect Hg(2+) ions with detection limit down to 4.3 × 10(-8) mol L(-1). The sensor developed in this work has a high potential for rapid and on-site detection of Hg(2+) ions. The sensor was successfully applied for assessment of Hg(2+) ions in real water samples collected from various locations of the Sindh River.

  17. 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.

  18. 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

  19. A novel fluoride ion colorimetric chemosensor based on coumarin

    NASA Astrophysics Data System (ADS)

    Zhuang, Xiaoqing; Liu, Weimin; Wu, Jiasheng; Zhang, Hongyan; Wang, Pengfei

    2011-09-01

    A novel visible colorimetric sensor ( L1) with high selectivity for fluoride ion based on coumarin has been synthesized by a simple modification of our earlier report. The chemosensor L1 shows an obvious color change from yellow to blue upon addition of fluoride ion with a large red shift of 145 nm in acetonitrile, and without interference of other anions such as Cl -, Br -, I -, NO 3-, H 2PO 4-, HSO 4-, and AcO -. The investigation of 1H NMR spectrum titration indicates the proposed mechanism is that F - first establishes a hydrogen bonding interaction with L1, and then the formation of [F-H-F] - induces deprotonation.

  20. 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

  1. 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...

  2. 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…

  3. 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.

  4. β-Galactosidase-based colorimetric paper sensor for determination of heavy metals.

    PubMed

    Hossain, S M Zakir; Brennan, John D

    2011-11-15

    We demonstrate a novel approach for rapid, selective, and sensitive detection of heavy metals using a solid-phase bioactive lab-on-paper sensor that is inkjet printed with sol-gel entrapped reagents to allow colorimetric visualization of the enzymatic activity of β-galactosidase (B-GAL). The bioactive paper assay is able to detect a range of heavy metals, either alone or as mixtures, in as little as 10 min, with detection limits as follows: Hg(II) = 0.001 ppm; Ag(I) = 0.002 ppm, Cu(II) = 0.020 ppm; Cd(II) = 0.020 ppm; Pb(II) = 0.140 ppm; Cr(VI) = 0.150 ppm; Ni(II) = 0.230 ppm. The paper-based assay was immune to interferences from nontoxic metal ions such as Na(+) or K(+), could be used to detect heavy metals that were spiked into tap water or lake water, and provided quantitative data that was in agreement with values obtained by atomic absorption. With the incorporation of standard chromogenic metal sensing reagents into a multiplexed bioactive paper sensor, it was possible to identify specific metals in mixtures, albeit with much lower detection limits than were obtained with the enzymatic assay. The paper-based sensor should be valuable for rapid, on-site screening of trace levels of heavy metals in resource limited areas and developing countries.

  5. 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.

  6. 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

  7. 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

  8. 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).

  9. 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

  10. 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

  11. 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

  12. [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

  13. 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

  14. 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.

  15. 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

  16. 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

  17. 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.

  18. A colorimetric probe based on desensitized ionene-stabilized gold nanoparticles for single-step test for sulfate ions

    NASA Astrophysics Data System (ADS)

    Arkhipova, Viktoriya V.; Apyari, Vladimir V.; Dmitrienko, Stanislava G.

    2015-03-01

    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.

  19. 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.

  20. 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

  1. 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.

  2. Visual & reversible sensing of cyanide in real samples by an effective ratiometric colorimetric probe & logic gate application.

    PubMed

    Bhardwaj, Shubhrajyotsna; Singh, Ashok Kumar

    2015-10-15

    A novel anion probe 3 (2,4-di-tert-butyl-6-((2(2,4-dinitrophenyl) hydrazono) methyl) phenol) has been unveiled as an effective ratiometric and colorimetric sensor for selective and rapid detection of cyanide. The sensing behavior was demonstrated by UV-vis experiments and NMR studies. This sensory system exhibited prominent visual color change toward cyanide ion over other testing anions in DMSO (90%) solvent, with a 1:1 binding stoichiometry and a detection limit down to 3.6×10(-8) mol L(-1). Sensor reveals specific anti-jamming activity and reversible in the presence of Cu(2+) ions. This concept has been applied to design a logic gate circuit at the molecular level. Further we developed coated graphite electrode using probe 3 as ionophore and studied the performance characteristics of electrode. The sensitivity of ratiometric-based colorimetric assay is below the 1.9 μM, accepted by the World Health Organization as the highest permissible cyanide concentration in drinking water. So it can be applied for both quantitative determination and qualitative supervising of cyanide concentrations in real samples.

  3. 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.

  4. Visual and highly sensitive detection of cancer cells by a colorimetric aptasensor based on cell-triggered cyclic enzymatic signal amplification.

    PubMed

    Zhang, Xianxia; Xiao, Kunyi; Cheng, Liwei; Chen, Hui; Liu, Baohong; Zhang, Song; Kong, Jilie

    2014-06-01

    Rapid and efficient detection of cancer cells at their earliest stages is one of the central challenges in cancer diagnostics. We developed a simple, cost-effective, and highly sensitive colorimetric method for visually detecting rare cancer cells based on cell-triggered cyclic enzymatic signal amplification (CTCESA). In the absence of target cells, hairpin aptamer probes (HAPs) and linker DNAs stably coexist in solution, and the linker DNA assembles DNA-AuNPs, producing a purple solution. In the presence of target cells, the specific binding of HAPs to the target cells triggers a conformational switch that results in linker DNA hybridization and cleavage by nicking endonuclease-strand scission cycles. Consequently, the cleaved fragments of linker DNA can no longer assemble into DNA-AuNPs, resulting in a red color. UV-vis spectrometry and photograph analyses demonstrated that this CTCESA-based method exhibited selective and sensitive colorimetric responses to the presence of target CCRF-CEM cells, which could be detected by the naked eye. The linear response for CCRF-CEM cells in a concentration range from 10(2) to 10(4) cells was obtained with a detection limit of 40 cells, which is approximately 20 times lower than the detection limit of normal AuNP-based methods without amplification. Given the high specificity and sensitivity of CTCESA, this colorimetric method provides a sensitive, label-free, and cost-effective approach for early cancer diagnosis and point-to-care applications.

  5. 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

  6. 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

  7. Loop-Mediated Isothermal Amplification for Rapid and Semiquantitative Detection of Loa loa Infection

    PubMed Central

    Drame, Papa M.; Fink, Doran L.; Kamgno, Joseph; Herrick, Jesica A.

    2014-01-01

    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

  8. Quantification of Endogenous Retinoids

    PubMed Central

    Kane, Maureen A.; Napoli, Joseph L.

    2014-01-01

    Numerous physiological processes require retinoids, including development, nervous system function, immune responsiveness, proliferation, differentiation, and all aspects of reproduction. Reliable retinoid quantification requires suitable handling and, in some cases, resolution of geometric isomers that have different biological activities. Here we describe procedures for reliable and accurate quantification of retinoids, including detailed descriptions for handling retinoids, preparing standard solutions, collecting samples and harvesting tissues, extracting samples, resolving isomers, and detecting with high sensitivity. Sample-specific strategies are provided for optimizing quantification. Approaches to evaluate assay performance also are provided. Retinoid assays described here for mice also are applicable to other organisms including zebrafish, rat, rabbit, and human and for cells in culture. Retinoid quantification, especially that of retinoic acid, should provide insight into many diseases, including Alzheimer’s disease, type 2 diabetes, obesity, and cancer. PMID:20552420

  9. Quantification of Secondary Metabolites.

    PubMed

    2016-01-01

    Plants are a rich source of secondary metabolites that have medicinal and aromatic properties. Secondary metabolites such as alkaloids, iridoids and phenolics generally produced by plants for their defence mechanisms have been implicated in the therapeutic properties of most medicinal plants. Hence, quantification of these metabolites will aid to discover new and effective drugs from plant sources and also to scientifically validate the existing traditional practices. Quantification of large group of phytochemicals such as phenolics and flavonoids is quantified in this context.

  10. Detection of biological thiols based on a colorimetric method*

    PubMed Central

    Xu, Yuan-yuan; Sun, Yang-yang; Zhang, Yu-juan; Lu, Chen-he; Miao, Jin-feng

    2016-01-01

    Biological thiols (biothiols), an important kind of functional biomolecules, such as cysteine (Cys) and glutathione (GSH), play vital roles in maintaining the stability of the intracellular environment. In past decades, studies have demonstrated that metabolic disorder of biothiols is related to many serious disease processes and will lead to extreme damage in human and numerous animals. We carried out a series of experiments to detect biothiols in biosamples, including bovine plasma and cell lysates of seven different cell lines based on a simple colorimetric method. In a typical test, the color of the test solution could gradually change from blue to colorless after the addition of biothiols. Based on the color change displayed, experimental results reveal that the percentage of biothiols in the embryonic fibroblast cell line is significantly higher than those in the other six cell lines, which provides the basis for the following biothiols-related study. PMID:27704750

  11. 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

  12. 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

  13. 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

  14. 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

  15. 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.

  16. Colorimetric aptasensor using unmodified gold nanoparticles for homogeneous multiplex detection.

    PubMed

    Niu, Shucao; Lv, Zhenzhen; Liu, Jinchuan; Bai, Wenhui; Yang, Shuming; Chen, Ailiang

    2014-01-01

    Colorimetric aptasensors using unmodified gold nanoparticles (AuNPs) have attracted much attention because of their low cost, simplicity, and practicality, and they have been developed for various targets in the past several years. However, previous research has focused on developing single-target assays. Here, we report the development of a homogeneous multiplex aptasensor by using more than one class of aptamers to stabilize AuNPs. Using sulfadimethoxine (SDM), kanamycin (KAN) and adenosine (ADE) as example targets, a KAN aptamer (750 nM), an SDM aptamer (250 nM) and an ADE aptamer (500 nM) were mixed at a 1∶1∶1 volume ratio and adsorbed directly onto the surface of unmodified AuNPs by electrostatic interaction. Upon the addition of any of the three targets, the conformation of the corresponding aptamer changed from a random coil structure to a rigid folded structure, which could not adsorb and stabilize AuNPs. The AuNPs aggregated in a specific reaction buffer (20 mM Tris-HCl containing 20 mM NaCl and 5 mM KCl), which led to a color change from red to purple/blue. These results demonstrate that the multiplex colorimetric aptasensor detected three targets simultaneously while maintaining the same sensitivity as a single-target aptasensor for each individual target. The multiplex aptasensor could be extended to other aptamers for various molecular detection events. Due to its simple design, easy operation, fast response, cost effectiveness and lack of need for sophisticated instrumentation, the proposed strategy provides a powerful tool to examine large numbers of samples to screen for a small number of potentially positive samples containing more than one analyte, which can be further validated using sophisticated instruments. PMID:25279730

  17. Bio-functionalized silver nanoparticles: a novel colorimetric probe for cysteine detection.

    PubMed

    Borase, Hemant P; Patil, Chandrashekhar D; Salunkhe, Rahul B; Suryawanshi, Rahul K; Kim, Beom S; Bapat, Vishwas A; Patil, Satish V

    2015-04-01

    Chemical interactions between nanoparticles and biomolecules are vital for applying nanoparticles in medicine and life science. Development of sensitive, rapid, low-cost, and eco-friendly sensors for the detection of molecules acting as disease indicator is need of an hour. In the present investigation, a green trend for silver nanoparticle synthesis was followed using leaf extract of Calotropis procera. Silver nanoparticles exhibited surface plasmon absorption peak at 421 nm, spherical shape with average size of 10 nm, and zeta potential of -22.4 mV. The as-synthesized silver nanoparticles were used for selective and sensitive detection of cysteine. Cysteine induces aggregation in stable silver nanoparticles owing to selective and strong interaction of -SH group of cysteine with silver nanoparticle surface. Cysteine-induced silver nanoparticle aggregation can be observed visually by change in color of silver nanoparticles from yellow to pink. Cysteine concentration was estimated colorimetrically by measuring absorption at surface plasmon wavelength. Limit of detection for cysteine using silver nanoparticles is ultralow, i.e., 100 nM. The mechanistic insight into cysteine detection by silver nanoparticles was investigated using FT-IR, TEM, DLS, and TLC analysis. Proposed method can be applied for the detection of cysteine in blood plasma and may give rise to a new insight into development of eco-friendly fabricated nanodiagnostic device in future.

  18. Ion-association method for the colorimetric determination of neomycin sulphate in pure and dosage forms

    NASA Astrophysics Data System (ADS)

    Amin, A. S.; Issa, Y. M.

    2003-03-01

    A simple, fairly rapid, sensitive and accurate method is described for the colorimetric determination of neomycin sulphate (NMS), based on the measurement of the absorbance of the extracted organic soluble ion-association complex formed between neomycin dictation and a bulky counter anion. Different chromotropic acid azo dyes were examined as counter ions. The effect of pH, the counter ion concentration, sequence of addition and solvents for extraction were also illustrated. The most suitable system is based on reagent VIII (pH 7.5) with chloroform as the extraction solvent. The use of other counter ions, in conjunction with their respective solvents, was found to be less sensitive. The neomycin-reagent VIII system exhibits negligible or no interference when used for the determination of up to 58 μg ml -1 of NMS in the presence of several drug excipiences. The method has been used for the determination of up to 58 μg ml -1 with a good recovery (99.8±1.5%), and the precision is supported by the low relative standard deviation ⩽1.35%. The sensitivity is discussed and the results are compared with the official method. The proposed method was applied successfully to the determination of NMS in pure and dosage forms, with a good precision and accuracy compared to the official one.

  19. 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

  20. A high-throughput colorimetric assay to measure the activity of glutamate decarboxylase.

    PubMed

    Yu, Kai; Hu, Sheng; Huang, Jun; Mei, Le-He

    2011-08-10

    A pH-sensitive colorimetric assay has been established to quantitatively measure glutamate decarboxylase (GAD) activity in bacterial cell extracts using a microplate format. GAD catalyzes the irreversible α-decarboxylation of L-glutamate to γ-aminobutyrate. The assay is based on the color change of bromocresol green due to an increase in pH as protons are consumed during the enzyme-catalyzed reaction. Bromocresol green was chosen as the indicator because it has a similar pK(a) to the acetate buffer used. The corresponding absorbance change at 620 nm was recorded with a microplate reader as the reaction proceeded. A difference in the enzyme preparation pH and optimal pH for GAD activity of 2.5 did not prevent this method from successfully allowing the determination of reaction kinetic parameters and the detection of improvements in enzymatic activity with a low coefficient of variance. Our assay is simple, rapid, requires minimal sample concentration and can be carried out in robotic high-throughput devices used as standard in directed evolution experiments. In addition, it is also applicable to other reactions that involve a change in pH.

  1. 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.

  2. 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

  3. 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

  4. 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

  5. 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.

  6. 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.

  7. 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.

  8. Detection of Salmonella by using the colorimetric DNA/rRNA sandwich hybridization in microtiter wells.

    PubMed

    Namimatsu, T; Tsuna, M; Imai, Y; Futo, S; Mitsuse, S; Sakano, T; Sato, S

    2000-06-01

    A rapid and readily available DNA probe kit was developed for the detection of Salmonella spp. This kit utilized the colorimetric DNA/rRNA sandwich hybridization method in microtiter wells. Within 3 hr Salmonella spp. in selective enrichment broth cultures were detected by the DNA probe kit. The kit effectively identified all of 187 strains of Salmonella tested and yielded no false-positive reactions in the examination of 674 pure cultures of non-salmonellae. The DNA probe kit could detect 10(5) cfu/ml in pure culture. A total of 379 naturally contaminated samples (raw chicken meat, liquid egg, animal feeds, poultry feces and frozen foods) were tested, both by the standard culture method and the DNA probe kit. The 169 of these samples were culture positive and 210 were culture negative. The sensitivity of the DNA probe kit was 98.2% (166/169) and the specificity was 99.5% (209/210). These results show that the DNA probe kit is a useful tool to examine a large number of various samples for contamination by Salmonella spp. in food and livestock industry. PMID:10907688

  9. Development of reverse transcription (RT)-PCR and real-time RT-PCR assays for rapid detection and quantification of viable yeasts and molds contaminating yogurts and pasteurized food products.

    PubMed

    Bleve, Gianluca; Rizzotti, Lucia; Dellaglio, Franco; Torriani, Sandra

    2003-07-01

    Reverse transcriptase PCR (RT-PCR) and real-time RT-PCR assays have been used to detect and quantify actin mRNA from yeasts and molds. Universal primers were designed based on the available fungal actin sequences, and by RT-PCR they amplified a specific 353-bp fragment from fungal species involved in food spoilage. From experiments on heat-treated cells, actin mRNA was a good indicator of cell viability: viable cells and cells in a nonculturable state were detected, while no signal was observed from dead cells. The optimized RT-PCR assay was able to detect 10 CFU of fungi ml(-1) in pure culture and 10(3) and 10(2) CFU ml(-1) in artificially contaminated yogurts and pasteurized fruit-derived products, respectively. Real-time RT-PCR, performed on a range of spoiled commercial food products, validated the suitability of actin mRNA detection for the quantification of naturally contaminating fungi. The specificity and sensitivity of the procedure, combined with its speed, its reliability, and the potential automation of the technique, offer several advantages to routine analysis programs that assess the presence and viability of fungi in food commodities.

  10. 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

  11. A novel colorimetric method for the detection of Escherichia coli using cytochrome c peroxidase-encoding bacteriophage.

    PubMed

    Hoang, Hoang A; Abe, Michiharu; Nakasaki, Kiyohiko

    2014-03-01

    A new rapid and simple method was developed for the detection of Escherichia coli by constructing a recombinant T4 phage carrying the cytochrome c peroxidase gene derived from Saccharomyces cerevisiae (T4ccp) using which, the colorimetric detection of E. coli K12 was examined. The oxidation activity toward the chromogenic substrate cytochrome c was demonstrated by the cytochrome c peroxidase (CCP) produced from the T4ccp genome. The color change caused by the oxidation of the substrate could be visually perceived. The possibility of interference in the detection by the coexistence of other bacteria was assessed using Pseudomonas aeruginosa as a nontarget bacterium, and it was confirmed that the coexistence of P. aeruginosa caused no interference in the detection of E. coli K12.

  12. Gum kondagogu reduced/stabilized silver nanoparticles as direct colorimetric sensor for the sensitive detection of Hg²⁺ in aqueous system.

    PubMed

    Rastogi, Lori; Sashidhar, R B; Karunasagar, D; Arunachalam, J

    2014-01-01

    A highly sensitive and selective method is reported for the colorimetric detection of Hg(2+) in aqueous system by using label free silver nanoparticles (Ag NPs). Ag NPs used in this method were synthesized by gum kondagogu (GK) which acted as both reducing and stabilizing agent. The average size of the GK-Ag NPs was found to be 5.0 ± 2.8 nm as revealed by transmission electron microscope (TEM) analysis and the nanoparticles were stable at various pH conditions (pH 4-11) and salt concentrations (5-100 mM). The GK reduced/stabilized Ag NPs (GK-Ag NPs) were directly used for the selective colorimetric reaction with Hg(2+) without any further modification. The bright yellow colour of Ag NPs was found to fade in a concentration dependent manner with the added Hg(+) ions. The fading response was directly correlated with increasing concentration of Hg(2+). More importantly, this response was found to be highly selective for Hg(2+) as the absorption spectra were found to be unaffected by the presence of other ions like; Na(+), K(+), Mg(2+), Ca(2+), Cu(2+), Ni(2+), Co(2+), As(3+), Fe(2+), Cd(2+), etc. The metal sensing mechanism is explained based on the turbidometric and X-ray diffraction (XRD) analysis of GK-Ag NPs with Hg(2+). The proposed method was successfully applied for the determination of Hg(2+) in various ground water samples. The reported method can be effectively used for the quantification of total Hg(2+) in samples, wherein the organic mercury is first oxidized to inorganic form by ultraviolet (UV) irradiation. The limit of quantification for Hg(2+) using the proposed method was as low as 4.9 × 10(-8) mol L(-1) (50 nM). The proposed method has potential application for on-field qualitative detection of Hg(2+) in aqueous environmental samples.

  13. 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

  14. 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...

  15. 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.

  16. A Simple and Green Route for Room-Temperature Synthesis of Gold Nanoparticles and Selective Colorimetric Detection of Cysteine.

    PubMed

    Bagci, Pelin Onsekizoglu; Wang, Yi-Cheng; Gunasekaran, Sundaram

    2015-09-01

    Gold nanoparticles (AuNPs) were synthesized at room temperature following a simple, rapid, and green route using fresh-squeezed apple juice as a reducing reagent. The optimal AuNPs, based on the particle color, stability, and color change suitable for colorimetric detection of cysteine (Cys), are synthesized using 5 mL of 10% apple juice, 1 mL of 10 mM gold precursor solution, and 1 mL of 0.1 M NaOH. Under this set of parameters, the AuNPs are synthesized within 30 min at room temperature. The average size (11.1 ± 3.2 nm) and ζ potential (-36.5 mV) of the AuNPs synthesized were similar to those of AuNPs prepared via the conventional citrate-reduction method. In the presence of Cys, unlike with any other amino acid, the AuNPs aggregated, possibly due to the gold-sulfur covalent interaction, yielding red-to-purple color change of the sample solution. The red-shift of the localized surface plasmon resonance peak of the AuNPs responsible for the color change was recorded by UV-vis spectrometer. The effect of other potential interferents such as glucose, ascorbic acid, K(+) , Na(+) , Ca(2+) , Zn(2+) , Ag(+) , Ni(2+) , Cu(2+) , Co(2+) , and Hg(2+) were also examined. The results show that AuNPs can be used to selectively detect and measure Cys with a linear dependency in the range of 2 to 100 μM and a limit of detection (signal-to-noise ratio > 3) of 50 nM. The results suggest that the green-synthesized AuNPs are useful for simple, rapid, and sensitive colorimetric detection of Cys, which is an essential amino acid in food and biological systems.

  17. 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

  18. [Colorimetric detection of human influenza A H1N1 virus by reverse transcription loop mediated isothermal amplification].

    PubMed

    Nie, Kai; Wang, Da-Yan; Qin, Meng; Gao, Rong-Bao; Wang, Miao; Zou, Shu-Mei; Han, Feng; Zhao, Xiang; Li, Xi-Yan; Shu, Yue-Long; Ma, Xue-Jun

    2010-03-01

    A simple, rapid and sensitive colorimetric Reverse Transcription Loop Mediated Isothermal Amplification (RT-LAMP) method was established to detect human influenza A H1N1 virus. The method employed a set of six specially designed primers that recognized eight distinct sequences of the HA gene for amplification of nucleic acid under isothermal conditions at 65 degrees C for one and half hour. The amplification process of RT-LAMP was monitored by the addition of HNB (Hydroxy naphthol blue) dye prior to amplification. A positive reaction was indicated by a color change from violet to sky blue and confirmed by agarose electrophoresis. The specificity of the RT-LAMP assay was validated by cross-reaction with different swine and human influenza virus including human seasonal influenza A /H1N1 A /H3N2, influenza B and swine A /H1N1. The sensitivity of this assay was evaluated by serial dilutions of RNA molecules from in vitro transcription of human influenza A H1N1 HA gene. The assay was further evaluated with 30 clinical specimens with suspected pandemic influenza A H1N1 virus infection in parallel with RT-PCR detection and 26 clinical specimens with seasonal influenza virus infection. Our results showed that the RT-LAMP was able to achieve a sensitivity of 60 RNA copies with high specificity, and detection rate was comparable to that of the RT-PCR with the clinical samples of pandemic influenza A H1N1 infection. The RT-LAMP reaction with HNB could also be measured at 650nm in a microplate reader for quantitative analysis. Thus, we concluded that this colorimetric RT-LAMP assay had potential for the rapid screening of the human influenza A H1N1 virus infection in National influenza monitoring network laboratories and sentinel hospitals of provincial and municipal region in China.

  19. 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

  20. 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

  1. Simultaneous, rapid, and sensitive quantification of 8-hydroxy-2'-deoxyguanosine and cotinine in human urine by on-line solid-phase extraction LC-MS/MS: correlation with tobacco exposure biomarkers NNAL.

    PubMed

    Chen, Chao-Yu; Jhou, Yu-Ting; Lee, Hui-Ling; Lin, Yu-Wen

    2016-09-01

    Cigarette smoke can increase oxidative DNA damage. The main component in cigarette smoke is nicotine. Nicotine is metabolized to cotinine, which can be regarded as a biomarker for measuring exposure to tobacco smoke. A sensitive, simple, and robust method based on on-line solid-phase extraction liquid chromatography with tandem mass spectrometry (on-line SPE LC-MS/MS) has been developed and validated for the simultaneous determination of 8-OHdG and cotinine. The matrix effects of 8-OHdG and cotinine were measured at 97.1 and 91.7 %, with values for CV at 4.4 and 4.2 %, respectively. The limits of detection of 8-OHdG and cotinine were 10.0 and 5.5 pg mL(-1), and the limits of quantification were 40.0 and 20.0 pg mL(-1), respectively. The total run time was 12 min. We quantified 8-OHdG and cotinine in the urine of 80 male subjects. The results showed the levels of 8-OHdG and cotinine in smokers were significantly higher than that in non-smokers. Furthermore, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol and its glucuronide conjugate (defined as total NNAL) are the nitrosation metabolites of nicotine. In this study, urinary levels of 8-OHdG and cotinine were well correlated with urinary levels of total NNAL. This is also the first study to focus on the future risk of oxidative stress from exposure to cigarette smoke based on the relationship between 8-OHdG levels, cotinine levels, and total NNAL concentrations in the urine of humans. Graphical Abstract On-line SPE LC-MS/MS for the simultaneous determination of 8-OHdG and cotinine in human urine. PMID:27422644

  2. 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

  3. A colorimetric method for determination of γ-glutamyl-S-ethenyl-cysteine in narbon vetch (Vicia narbonensis L.) seeds.

    PubMed

    Sánchez-Vioque, R; Rodríguez-Conde, M F; Vioque, J; Girón-Calle, J; Santana-Méridas, O; De-los-Mozos-Pascual, M; Izquierdo-Melero, M E; Alaiz, M

    2011-11-15

    A new colorimetric method based on the bleaching of the iodoplatinate ion has been developed for fast and easy determination of γ-glutamyl-S-ethenyl-cysteine (GEC) in narbon vetch (Vicia narbonensis L.) seeds. The calibration curve showed a good correlation (r(2)=0.9959) between absorbance and GEC amounts from 5.5 to 33 μg (10-59.78 μmol/L). The limits of detection and quantification were 1.16 and 3.55 μmol/L, respectively, and no significant interferences from other sulfur-containing compounds were observed. The method showed excellent repeatability (relative standard deviation [RSD]=0.28%), reproducibility (RSD=4.4%), and accuracy (94%). Determination of GEC in 20 narbon vetch accessions yielded values that were in agreement with those reported previously using capillary electrophoresis and high-performance liquid chromatography methods. The method could be especially valuable for determination of GEC during the process of production of new low-GEC narbon vetch varieties.

  4. New Chitosan-Thiomer: An Efficient Colorimetric Sensor and Effective Sorbent for Mercury at Ultralow Concentration.

    PubMed

    Chauhan, Kalpana; Singh, Prem; Singhal, Rakesh Kumar

    2015-12-01

    This paper describes an innovative procedure for the fabrication of a facile colorimetric sensor in one step with thiol functional group for Hg(2+) detection at trace level. The sensor was successfully synthesized via chitosan isothiouronium salt intermediate with innocuous low cost thiourea reagent under microwave irradiation. It is an innovative green approach to achieve thiol functionalization with a high degree of substitution. Thiomer was characterized by titrimetry, FTIR, (1)H NMR, elemental analysis (CHNS), and EDX for extent of modification with detail structure. The synthesized and well characterized thiomer was screened for sensor application. The sensing solution of thiomer resulted in an instantaneous sharp color change from colorless, yellow, to brown with increase in Hg(2+) concentration. Chitosan thiomer also exhibited high sensitivity and selectivity for Hg(2+) over other possible interfering ions in aqueous media. The sensing responses were visualized quantitatively with quick response, good selectivity, high sensitivity, and a low detection limit of ∼0.465 ppb by the naked eye. The same was tested with a paper strip method for technological applications. Furthermore, the as-prepared sensors also exhibited exceptional sorption potential for Hg(2+) even from ultralow concentration aqueous solution and reduced the Hg(2+) concentration from 10 ppb to the extremely low level of ∼0.04 ppb as studied by cyclic voltammetry. Thus, the proposed method is simple, promising, and rapid without any complicated modifying step and is an economical alternative to traditional Hg(2+) sensors for rapid sensor application in environmental water samples at ppb levels.

  5. 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.

  6. Quantification of Secondary Metabolites.

    PubMed

    2016-01-01

    Plants are a rich source of secondary metabolites that have medicinal and aromatic properties. Secondary metabolites such as alkaloids, iridoids and phenolics generally produced by plants for their defence mechanisms have been implicated in the therapeutic properties of most medicinal plants. Hence, quantification of these metabolites will aid to discover new and effective drugs from plant sources and also to scientifically validate the existing traditional practices. Quantification of large group of phytochemicals such as phenolics and flavonoids is quantified in this context. PMID:26939265

  7. 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

  8. 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.

  9. Colorimetric Determination of Color of Aerial Mycelium of Streptomycetes1

    PubMed Central

    Lyons, Allister J.; Pridham, Thomas G.

    1965-01-01

    Lyons, Allister J., Jr. (Northern Regional Research Laboratory, Peoria, Ill.) and Thomas G. Pridham. Colorimetric determination of color of aerial mycelium of streptomycetes. J. Bacteriol. 89:159–169. 1965.—For some time, streptomycete taxonomists have been seeking to describe more accurately the colors of aerial mycelium. Some of the descriptive systems involve many different color names and groups. Others combine many colors into a few groups. All the systems and methods leave much to be desired. To obtain an accurate description, a colorimeter with a reflectance attachment was used to examine streptomycete aerial mycelium of 37 strains, representing all of the major aerial mycelium color groups. Each color was characterized by three values: dominant wavelength in millimicrons, and purity and brightness in percentages. All colors of aerial mycelium were of low purity (< 25%). Most of the dominant wavelengths were in the yellow to yellow-green bands of the spectrum. Most of the color tabs matched visually with the streptomycete strains had purities of a higher value than those of the cultures. The reflectance instrument seems to allow an objective description, and its use may help to clarify the color problem with streptomycetes. It is concluded that present color descriptions are inadequate and that the significance of color in speciation requires critical examination. PMID:14255657

  10. Colorimetric determination of reducing normality in the Purex process

    SciTech Connect

    Baumann, E.W.

    1983-07-01

    Adjustment of the valence state of plutonium from extractable Pu(IV) to nonextractable Pu(III) in the Purex process is accomplished by addition of reductants such as Fe(II), hydroxylamine nitrate (HAN), or U(IV). To implement on-line monitoring of this reduction step for improved process control at the Savannah River Plant, a simple colorimetric method for determining excess reductant (reducing normality) was developed. The method is based on formation of a colored complex of Fe(II) with FerroZine (Hach Chemical Company). The concentration of Fe(II) is determined directly. The concentration of HAN or U(IV), in addition to Fe(II), is determined indirectly as Fe(II), produced through reduction of Fe(III). Experimental conditions for a HAN-Fe(III) reaction of known stoichiometry were established. The effect of hydrazine, which stabilizes U(IV), was also determined. Real-time measurements of color development were made that simulated on-line performance. A laboratory analytical procedure is included. 5 references, 8 figures.

  11. 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.

  12. 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.

  13. [Rapid antibiotic susceptibility test in Clinical Microbiology].

    PubMed

    March Rosselló, Gabriel Alberto; Bratos Pérez, Miguel Ángel

    2016-01-01

    The most widely used antibiotic susceptibility testing methods in Clinical Microbiology are based on the phenotypic detection of antibiotic resistance by measuring bacterial growth in the presence of the antibiotic being tested. These conventional methods take typically 24hours to obtain results. A review is presented here of recently developed techniques for the rapid determination of antibiotic susceptibility. Data obtained with different methods such as molecular techniques, flow cytometry, chemiluminescence, mass spectrometry, commercial methods used in routine work, colorimetric methods, nephelometry, microarrays, microfluids, and methods based on cell disruption and sequencing, are analyzed and discussed in detail.

  14. Rapid detection of melamine in whole milk mediated by unmodified gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Wei, Fang; Lam, Robert; Cheng, Stacy; Lu, Steven; Ho, Dean; Li, Na

    2010-03-01

    A strategy for rapid and facile detection of melamine in food and other substances is described. Unmodified gold nanoparticles enable a colorimetric signal output that is amplified through integration by precipitation formation via a chemical component specific to melamine. The sensitivity was determined to be ˜40 ppb with a two standard deviation cutoff.

  15. Highly efficient colorimetric detection of target cancer cells utilizing superior catalytic activity of graphene oxide-magnetic-platinum nanohybrids

    NASA Astrophysics Data System (ADS)

    Kim, Moon Il; Kim, Min Su; Woo, Min-Ah; Ye, Youngjin; Kang, Kyoung Suk; Lee, Jinwoo; Park, Hyun Gyu

    2014-01-01

    Enzyme-linked immunosorbent assays (ELISAs) have most widely been applied in immunoassays for several decades. However, several unavoidable limitations (e.g., instability caused by structural unfolding) of natural enzymes have hindered their widespread applications. Here, we describe a new nanohybrid consisting of Fe3O4 magnetic nanoparticles (MNPs) and platinum nanoparticles (Pt NPs), simultaneously immobilized on the surface of graphene oxide (GO). By synergistically integrating highly catalytically active Pt NPs and MNPs on GO whose frameworks possess high substrate affinity, the nanohybrid is able to achieve up to a 30-fold higher maximal reaction velocity (Vmax) compared to that of free GO for the colorimetric reaction of the peroxidase substrate, 3,3',5,5'-tetramethylbenzidine (TMB), and enable rapid detection of target cancer cells. Specifically, using this new assay system, clinically important breast cancer cells are detected in a 5 min time period at room temperature with high specificity and sensitivity. The remarkably high capability to catalyze oxidation reactions could allow the nanohybrid to replace conventional peroxidase-based immunoassay systems as part of new, rapid, robust and convenient assay systems which can be widely utilized for the identification of important target molecules.Enzyme-linked immunosorbent assays (ELISAs) have most widely been applied in immunoassays for several decades. However, several unavoidable limitations (e.g., instability caused by structural unfolding) of natural enzymes have hindered their widespread applications. Here, we describe a new nanohybrid consisting of Fe3O4 magnetic nanoparticles (MNPs) and platinum nanoparticles (Pt NPs), simultaneously immobilized on the surface of graphene oxide (GO). By synergistically integrating highly catalytically active Pt NPs and MNPs on GO whose frameworks possess high substrate affinity, the nanohybrid is able to achieve up to a 30-fold higher maximal reaction velocity (Vmax

  16. 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.

  17. 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

  18. 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

  19. 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.

  20. The proteomics quantification dilemma.

    PubMed

    Jungblut, Peter R

    2014-07-31

    Proteomics is dominated today by the protein expression discourse, which favorites the bottom-up approach because of its high throughput and its high sensitivity. For quantification this proceeding is misleading, if a protein is present with more than one protein species in the sample to be analyzed. The protein speciation discourse considers this more realistic situation and affords the top-down procedures or at least a separation of the protein species in advance to identification and quantification. Today all of the top-down procedures are one order of magnitude less sensitive than the bottom-up ones. To increase sensitivity and to increase throughput are major challenges for proteomics of the next years. This article is part of a Special Issue entitled: 20years of Proteomics in memory of Viatliano Pallini. Guest Editors: Luca Bini, Juan J. Calvete, Natacha Turck, Denis Hochstrasser and Jean-Charles Sanchez. PMID:24681132

  1. Development of visible diode-laser/fiber-optic colorimetric system for the simple and inexpensive quantification of a protein

    NASA Astrophysics Data System (ADS)

    Kim, SungHo; Noh, Young S.; Byun, Gill S.; Jang, Sung-Keun

    1998-04-01

    A simple and inexpensive quantitative detection of protein in one droplet solution was achieved by a portable diode-laser- based-optical-fiber absorption spectrometry. A system consisted of a visible diode laser, a photodiode and a pair of optical fibers. The conventional Lowry method was used to measure the blue-colored protein solutions. In this study, one droplet of sample, which filled up the void between the end surfaces of two optical fibers, served as a sample cell. The volume of a droplet was 50 (mu) L, which is 1/100 of the cuvette of UV/VIS spectrometer cell used in Lowry method. Although the required sample volume decreased up to 100 times, the detection range was comparable to that of conventional Lowry method in the range of g/L - mg/L.

  2. 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.

  3. Formation of colorimetric fingerprints on nano-patterned deterministic aperiodic surfaces.

    PubMed

    Boriskina, Svetlana V; Lee, Sylvanus Y K; Amsden, Jason J; Omenetto, Fiorenzo G; Dal Negro, Luca

    2010-07-01

    Periodic gratings and photonic bandgap structures have been studied for decades in optical technologies. The translational invariance of periodic gratings gives rise to well-known angular and frequency filtering of the incident radiation resulting in well-defined scattered colors in response to broadband illumination. Here, we demonstrate the formation of highly complex structural color patterns, or colorimetric fingerprints, in two-dimensional (2D) deterministic aperiodic gratings using dark field scattering microscopy. The origin of colorimetric fingerprints is explained by rigorous full-wave numerical simulations based on the generalized Mie theory. We show that unlike periodic gratings, aperiodic nanopatterned surfaces feature a broadband frequency response with wide angular intensity distributions governed by the distinctive Fourier properties of the aperiodic structures. Finally, we will discuss a range of potential applications of colorimetric fingerprints for optical sensing and spectroscopy. PMID:20639942

  4. Highly efficient colorimetric detection of target cancer cells utilizing superior catalytic activity of graphene oxide-magnetic-platinum nanohybrids.

    PubMed

    Kim, Moon Il; Kim, Min Su; Woo, Min-Ah; Ye, Youngjin; Kang, Kyoung Suk; Lee, Jinwoo; Park, Hyun Gyu

    2014-01-01

    Enzyme-linked immunosorbent assays (ELISAs) have most widely been applied in immunoassays for several decades. However, several unavoidable limitations (e.g., instability caused by structural unfolding) of natural enzymes have hindered their widespread applications. Here, we describe a new nanohybrid consisting of Fe₃O₄ magnetic nanoparticles (MNPs) and platinum nanoparticles (Pt NPs), simultaneously immobilized on the surface of graphene oxide (GO). By synergistically integrating highly catalytically active Pt NPs and MNPs on GO whose frameworks possess high substrate affinity, the nanohybrid is able to achieve up to a 30-fold higher maximal reaction velocity (V(max)) compared to that of free GO for the colorimetric reaction of the peroxidase substrate, 3,3',5,5'-tetramethylbenzidine (TMB), and enable rapid detection of target cancer cells. Specifically, using this new assay system, clinically important breast cancer cells are detected in a 5 min time period at room temperature with high specificity and sensitivity. The remarkably high capability to catalyze oxidation reactions could allow the nanohybrid to replace conventional peroxidase-based immunoassay systems as part of new, rapid, robust and convenient assay systems which can be widely utilized for the identification of important target molecules.

  5. 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.

  6. Simple, field portable colorimetric detection device for organic peroxides and hydrogen peroxide

    SciTech Connect

    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.

  7. 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

  8. A boron-dipyrromethene-based fluorescent probe for colorimetric and ratiometric detection of sulfite.

    PubMed

    Gu, Xianfeng; Liu, Chunhua; Zhu, Yi-Chun; Zhu, Yi-Zhun

    2011-11-23

    BODIPY-Le, a colorimetric and ratiometric fluorescent probe based on boron-dipyrromethene for selective detection sulfite ion, was investigated. Boron-dipyrromethene levulinyl ester (BODIPY-Le) is composed of an indole-based BODIPY dye and the levulinyl protective group, which could be easily and selectively deprotected by sulfites. As a result, the absorption and emission spectra show a dramatic red shift, and the development of a colorimetric and ratiometric fluorescent sulfite probe could be achieved. Besides, BODIPY-Le also exhibited prominent turn-on or turn-off type fluorogenic signaling toward sulfite ions once excited at 510 and 620 nm, respectively.

  9. SERS Quantification of Entacapone Isomers

    NASA Astrophysics Data System (ADS)

    Marković, Marina; Biljan, Tomislav

    2010-08-01

    Raman spectroscopy, due to its non-destructive character and speed, has found widespread use in pharmaceutical applications [1]. It is also being used for quantifying various isomer mixtures, best known being the quantification of xylene isomers [2-3]. Solid-state isomer quantification of entacapone was earlier reported [4]. Here, we report quantification of isomer mixture of an active pharmaceutical substance, in solution, by SERS.

  10. Protein quantification using a cleavable reporter peptide.

    PubMed

    Duriez, Elodie; Trevisiol, Stephane; Domon, Bruno

    2015-02-01

    Peptide and protein quantification based on isotope dilution and mass spectrometry analysis are widely employed for the measurement of biomarkers and in system biology applications. The accuracy and reliability of such quantitative assays depend on the quality of the stable-isotope labeled standards. Although the quantification using stable-isotope labeled peptides is precise, the accuracy of the results can be severely biased by the purity of the internal standards, their stability and formulation, and the determination of their concentration. Here we describe a rapid and cost-efficient method to recalibrate stable isotope labeled peptides in a single LC-MS analysis. The method is based on the equimolar release of a protein reference peptide (used as surrogate for the protein of interest) and a universal reporter peptide during the trypsinization of a concatenated polypeptide standard. The quality and accuracy of data generated with such concatenated polypeptide standards are highlighted by the quantification of two clinically important proteins in urine samples and compared with results obtained with conventional stable isotope labeled reference peptides. Furthermore, the application of the UCRP standards in complex samples is described.

  11. Protein quantification using a cleavable reporter peptide.

    PubMed

    Duriez, Elodie; Trevisiol, Stephane; Domon, Bruno

    2015-02-01

    Peptide and protein quantification based on isotope dilution and mass spectrometry analysis are widely employed for the measurement of biomarkers and in system biology applications. The accuracy and reliability of such quantitative assays depend on the quality of the stable-isotope labeled standards. Although the quantification using stable-isotope labeled peptides is precise, the accuracy of the results can be severely biased by the purity of the internal standards, their stability and formulation, and the determination of their concentration. Here we describe a rapid and cost-efficient method to recalibrate stable isotope labeled peptides in a single LC-MS analysis. The method is based on the equimolar release of a protein reference peptide (used as surrogate for the protein of interest) and a universal reporter peptide during the trypsinization of a concatenated polypeptide standard. The quality and accuracy of data generated with such concatenated polypeptide standards are highlighted by the quantification of two clinically important proteins in urine samples and compared with results obtained with conventional stable isotope labeled reference peptides. Furthermore, the application of the UCRP standards in complex samples is described. PMID:25411902

  12. 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

  13. 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

  14. 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.

  15. 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.

  16. 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.

  17. 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

  18. 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

  19. 3D graphene network@WO3 nanowire composites: a multifunctional colorimetric and electrochemical biosensing platform.

    PubMed

    Ma, Ye; Zhao, Minggang; Cai, Bin; Wang, Wei; Ye, Zhizhen; Huang, Jingyun

    2014-10-01

    A three dimensional graphene network (3DGN)@WO3 nanowire (NW) sensor is proposed which can perform colorimetric and electrochemical sensing techniques to detect H2O2, ascorbic acid and dopamine. The 3DGN provides three functions: anchoring, separating, conducting, while the WO3 NWs maximize surface area and catalyse reactions.

  20. Colorimetric Detection of Creatinine Based on Plasmonic Nanoparticles via Synergistic Coordination Chemistry.

    PubMed

    Du, Jianjun; Zhu, Bowen; Leow, Wan Ru; Chen, Shi; Sum, Tze Chien; Peng, Xiaojun; Chen, Xiaodong

    2015-09-01

    A simple and portable colorimetric assay for creatinine detection is fabricated based on the synergistic coordination of creatinine and uric acid with Hg(2+) on the surface of gold nanoparticles, which exhibits good selectivity and sensitivity. Point-of-care clinical creatinine monitoring can be supported for monitoring renal function and diagnosing corresponding renal diseases at home.

  1. Colorimetric sensor strips for formaldehyde assay utilizing fluoral-p decorated polyacrylonitrile nanofibrous membranes.

    PubMed

    Wang, Xueqin; Si, Yang; Mao, Xue; Li, Yan; Yu, Jianyong; Wang, Huaping; Ding, Bin

    2013-09-01

    A facile, ultrasensitive, and selective sensor strip utilizing 4-amino-3-penten-2-one (fluoral-p) functionalized electrospun polyacrylonitrile (PAN) (PAN/fluoral-p) nanofibrous membranes has been successfully developed for naked-eye colorimetric assay of formaldehyde. The sensor strips presented a significant reflectance decreasing band at 417 nm which induced a vivid color change from white to yellow and achieved a much lower naked-eye detection limit of 40 ppb compared with the World Health Organization standard (80 ppb). Based on the specific Hantzsch reaction between fluoral-p and formaldehyde, the as-prepared PAN/fluoral-p membranes are highly selective to formaldehyde with little interference from other volatile organic compounds and the proposed mechanism of this reaction is discussed carefully. Moreover, the colorimetric responses were visually quantitative using UV-vis spectra and the color difference calculated from L*, a*, b* values. Furthermore, due to the extremely large surface area and high porosity of the as-spun PAN nanofibrous membranes, the sensitivity of the nanofibrous membranes-based strips is much higher than traditional filter paper-based ones. Hence, such promising portable colorimetric sensor strips could not only potentially allow for assaying gaseous formaldehyde, but also facilitate the design and development of a novel colorimetric sensing system based on nanofibrous membranes. PMID:23831600

  2. Assessment of colorimetric amplification methods in a paper-based immunoassay for diagnosis of malaria.

    PubMed

    Lathwal, Shefali; Sikes, Hadley D

    2016-04-21

    Colorimetric detection methods that produce results readable by eye are important for diagnostic tests in resource-limited settings. In this work, we have compared three main types of colorimetric methods - enzymatic reactions, silver deposition catalyzed by gold nanoparticles, and polymerization-based amplification - in a paper-based immunoassay for detection of Plasmodium falciparum histidine-rich protein 2, a biomarker of malarial infection. We kept the binding events in the immunoassay constant in order to isolate the effect of the detection method on the outcome of the test. We have highlighted that the optimal readout time in a test can vary significantly - ranging from immediately after addition of a visualization agent to 25 minutes after addition of a visualization agent - depending on the colorimetric method being used, and accurate time keeping is essential to prevent false positives in methods where substantial color develops over time in negative tests. We have also shown that the choice of a colorimetric method impacts the calculated limit-of-detection, the ease of visual perception of the readout, and the total cost of the assay, and therefore directly impacts the feasibility and the ease-of-use of a test in field settings.

  3. A highly sensitive gold nanoparticle-based colorimetric probe for pyrophosphate using a competition assay approach.

    PubMed

    Kim, Sudeok; Eom, Min Sik; Kim, Seung Kyung; Seo, Seong Hyeok; Han, Min Su

    2013-01-01

    In this study, a mixture of [Zn(2)(1,3-bis[bis(2-pyridylmethyl)aminomethyl]benzene)](4+) ([Zn(2)(BBPAB)](4+)) and 11-mercaptoundecylphosphoric acid functionalized gold nanoparticles (Phos-AuNPs) is shown to be a highly sensitive colorimetric probe that can easily detect pyrophosphate (PPi) at less than 200 nM with the naked eye.

  4. Single Chain Fragment Variable Recombinant Antibody Functionalized Gold Nanoparticles for a Highly Sensitive Colorimetric Immunoassay

    PubMed Central

    Liu, Yang; Liu, Yi; Raymond, Raymond L.; Zeng, Xiangqun

    2009-01-01

    In this report, the peptide linker connecting scFv VH and VL domains were genetically modified to contain different amino acids (i.e. cysteine (scFv-cys) or histidines ( scFv-his)) to enable the scFv to adsorb or self-assemble onto the gold nanoparticles (NPs). The scFv-cys stabilized gold NPs were used to develop a highly sensitive colorimetric immunosensor. The scFv-cys stabilized gold NPs were characterized by UV-vis spectra, transmission electron microscope (TEM) and FT-IR. After adding the antigen rabbit IgG, the solution of scFv-cys stabilized gold NPs shows obvious visible color change from deep red to light purple due to the aggregation of the gold nanoparticles. Based on the colorimetric aggregation of scFv-cys stabilized gold NPs, the immunosensor exhibits high sensitivity with detection limit of 1.7 nM and good specificity. The good properties of the colorimetric aggregation immunosensor would be attributed to the small size of scFv and the covalent link between the scFv and gold NPs that improve the better orientation and enhance the probe density. With the advantages of speed, simplicity and specificity, the colorimetric immunoassay based on the functionalized scFv stabilized gold NPs represents a promising approach for protein analysis and clinical diagnostics. PMID:19327975

  5. 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.

  6. 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.

  7. Iodine-Mediated Etching of Gold Nanorods for Plasmonic ELISA Based on Colorimetric Detection of Alkaline Phosphatase.

    PubMed

    Zhang, Zhiyang; Chen, Zhaopeng; Wang, Shasha; Cheng, Fangbin; Chen, Lingxin

    2015-12-23

    Here, we propose a plasmonic enzyme-linked immunosorbent assay (ELISA) based on highly sensitive colorimetric detection of alkaline phosphatase (ALP), which is achieved by iodine-mediated etching of gold nanorods (AuNRs). Once the sandwich-type immunocomplex is formed, the ALP bound on the polystyrene microwells will hydrolyze ascorbic acid 2-phosphate into ascorbic acid. Subsequently, iodate is reduced to iodine, a moderate oxidant, which etches AuNRs from rod to sphere in shape. The shape change of AuNRs leads to a blue-shift of longitudinal localized surface plasmon resonance. As a result, the solution of AuNRs changes from blue to red. Benefiting from the highly sensitive detection of ALP, the proposed plasmonic ELISA has achieved an ultralow detection limit (100 pg/mL) for human immunoglobulin G (IgG). Importantly, the visual detection limit (3.0 ng/mL) allows the rapid differential diagnosis with the naked eye. The further detection of human IgG in fetal bovine serum indicates its applicability to the determination of low abundance protein in complex biological samples.

  8. 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.

  9. 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.

  10. 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

  11. A universal probe design for colorimetric detection of single-nucleotide variation with visible readout and high specificity.

    PubMed

    Chen, Xueping; Zhou, Dandan; Shen, Huawei; Chen, Hui; Feng, Wenli; Xie, Guoming

    2016-02-02

    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.

  12. A colorimetric assay for high-throughput screening of inhibitors of herpes simplex virus type 1 alkaline nuclease.

    PubMed

    Bronstein, J C; Weber, P C

    2001-06-15

    Herpes simplex virus type 1 (HSV-1) encodes a deoxyribonuclease that is frequently referred to as alkaline nuclease (AN) because of its elevated pH optimum. Studies with recombinant viruses which contain deletions in the HSV-1 gene encoding AN have indicated that this enzyme is required for efficient virus replication and therefore represents a potential target for novel antiviral therapies. A simple colorimetric assay for deoxyribonuclease activity employing a DNA-methyl green substrate was adapted for use in a high-throughput screen to identify small molecule inhibitors of this enzyme. This screen identified 1,2-benzoisothiazolin-3-one as a specific inhibitor of AN, since it exhibited activity against AN but was completely inactive against bovine pancreatic DNaseI. Subsequent studies revealed that this compound most likely inhibited AN by forming disulfide linkages with one or more exposed cysteine residues on the surface of the enzyme and that AN was sensitive to sulfhydryl-group-modifying reagents in general. These results demonstrated the utility of this DNA-methyl green substrate-based assay in both the rapid identification and the characterization of novel small molecule inhibitors of the AN encoded by HSV-1 and other herpesviruses.

  13. 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

  14. 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.

  15. 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

  16. Ultrasensitive, colorimetric detection of microRNAs based on isothermal exponential amplification reaction-assisted gold nanoparticle amplification.

    PubMed

    Li, Ru-Dong; Yin, Bin-Cheng; Ye, Bang-Ce

    2016-12-15

    MicroRNAs (miRNAs) play important roles in a wide range of biological processes, and their aberrant expressions are linked to a large number of human diseases and disorders. In this work, we developed a colorimetric method for rapid, ultrasensitive miRNA detection via isothermal exponential amplification reaction (EXPAR)-assisted gold nanoparticle (AuNP) amplification. The sensing probe designed with a tandem phosphorothioate modification in the backbone of the polyadenines at the 5' terminus was employed to directly assemble onto the surface of AuNP with high adsorption affinity. The recognition domain at the 3' terminus of the sensing probe hybridizes with target miRNAs to trigger EXPAR with exponential signal amplification. With the amplification reaction with the action of DNA polymerase, the sensing probe gradually detaches from the AuNP, resulting in the aggregation of bare AuNPs in the high-salt reaction environment due to lack of DNA protection. The presence of AuNP aggregation is conveniently measured by UV-vis spectroscopy. Our proposed method could provide a linear detection range from 50fM to 10nM with a detection limit of ∼46fM within 60min, and also discriminate a single-nucleotide difference between homologous miRNAs. PMID:27498329

  17. 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

  18. 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.

  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. Employment of colorimetric enzyme assay for monitoring expression and solubility of GST fusion proteins targeted to inclusion bodies.

    PubMed

    Mačinković, Igor S; Abughren, Mohamed; Mrkic, Ivan; Grozdanović, Milica M; Prodanović, Radivoje; Gavrović-Jankulović, Marija

    2013-12-01

    High levels of recombinant protein expression can lead to the formation of insoluble inclusion bodies. These complex aggregates are commonly solubilized in strong denaturants, such as 6-8M urea, although, if possible, solubilization under milder conditions could facilitate subsequent refolding and purification of bioactive proteins. Commercially available GST-tag assays are designed for quantitative measurement of GST activity under native conditions. GST fusion proteins accumulated in inclusion bodies are considered to be undetectable by such assays. In this work, solubilization of recombinantly produced proteins was performed in 4M urea. The activity of rGST was assayed in 2M urea and it was shown that rGST preserves 85% of its activity under such denaturing conditions. A colorimetric GST activity assay with 1-chloro-2, 4-dinitrobenzene (CDNB) was examined for use in rapid detection of expression targeted to inclusion bodies and for the identification of inclusion body proteins which can be solubilized in low concentrations of chaotropic agents. Applicability of the assay was evaluated by tracking protein expression of two GST-fused allergens of biopharmaceutical value in E. coli, GST-Der p 2 and GST-Mus a 5, both targeted to inclusion bodies.

  1. 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

  2. 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.

  3. 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

  4. 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.

  5. 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.

  6. 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.

  7. 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

  8. Functionalized magnetic microparticle-based colorimetric platform for influenza A virus detection.

    PubMed

    Chen, Chaohui; Zou, Zhong; Chen, Lu; Ji, Xinghu; He, Zhike

    2016-10-28

    A colorimetric platform for influenza A virus detection was developed by using the high efficiency of enzymatic catalysis and the reduction of gold ions with hydrogen peroxide. Aptamer-functionalized magnetic microparticles were synthesized to capture the influenza A virus. This was followed by the binding of ConA-GOx-AuNPs to the H3N2 virus through the ConA-glycan interaction. The sandwich complex was subsequently dispersed in glucose solution to trigger an enzymatic reaction to produce hydrogen peroxide, which controlled the growth of gold nanoparticles and produced colored solutions. The determination of H3N2 concentration was realized by comparing the two differently colored gold nanoparticles. This method could detect the target virus as low as 11.16 μg ml(-1). Furthermore, it opens new opportunities for sensitive and colorimetric detection of viruses and proteins. PMID:27655150

  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.

  10. Functionalized magnetic microparticle-based colorimetric platform for influenza A virus detection.

    PubMed

    Chen, Chaohui; Zou, Zhong; Chen, Lu; Ji, Xinghu; He, Zhike

    2016-10-28

    A colorimetric platform for influenza A virus detection was developed by using the high efficiency of enzymatic catalysis and the reduction of gold ions with hydrogen peroxide. Aptamer-functionalized magnetic microparticles were synthesized to capture the influenza A virus. This was followed by the binding of ConA-GOx-AuNPs to the H3N2 virus through the ConA-glycan interaction. The sandwich complex was subsequently dispersed in glucose solution to trigger an enzymatic reaction to produce hydrogen peroxide, which controlled the growth of gold nanoparticles and produced colored solutions. The determination of H3N2 concentration was realized by comparing the two differently colored gold nanoparticles. This method could detect the target virus as low as 11.16 μg ml(-1). Furthermore, it opens new opportunities for sensitive and colorimetric detection of viruses and proteins.

  11. 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.

  12. 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

  13. G-quadruplex-generating PCR for visual colorimetric detection of amplicons

    PubMed Central

    Bhadra, Sanchita; Codrea, Vlad; Ellington, Andrew D.

    2013-01-01

    We have developed a self-reporting PCR system for visual colorimetric gene detection and distinction of single nucleotide polymorphisms (SNP). 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 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

  14. Comparative analysis of colorimetric staining in skin using open-source software.

    PubMed

    Billings, Paul C; Sanzari, Jenine K; Kennedy, Ann R; Cengel, Keith A; Seykora, John T

    2015-02-01

    Colorimetric staining techniques such as immunohistochemistry (IHC), immunofluorescence (IF) and histochemistry (HC) provide useful information regarding the localization and relative amount of a molecule/substance in skin. We have developed a novel, straightforward method to assess colorimetric staining by combining features from two open-source software programs. As a proof of principle, we demonstrate the utility of this approach by analysing changes in skin melanin deposition during the radiation-induced tanning response of Yucatan mini-pigs. This method includes a visualization step to validate the accuracy of colour selection before quantitation to ensure accuracy. The data show that this method is robust and will provide a means to obtain accurate comparative analyses of staining in IHC/IF/HC samples.

  15. 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.

  16. 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

  17. 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.

  18. 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.

  19. 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.

  20. 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

  1. 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

  2. Functionalized magnetic microparticle-based colorimetric platform for influenza A virus detection

    NASA Astrophysics Data System (ADS)

    Chen, Chaohui; Zou, Zhong; Chen, Lu; Ji, Xinghu; He, Zhike

    2016-10-01

    A colorimetric platform for influenza A virus detection was developed by using the high efficiency of enzymatic catalysis and the reduction of gold ions with hydrogen peroxide. Aptamer-functionalized magnetic microparticles were synthesized to capture the influenza A virus. This was followed by the binding of ConA-GOx-AuNPs to the H3N2 virus through the ConA-glycan interaction. The sandwich complex was subsequently dispersed in glucose solution to trigger an enzymatic reaction to produce hydrogen peroxide, which controlled the growth of gold nanoparticles and produced colored solutions. The determination of H3N2 concentration was realized by comparing the two differently colored gold nanoparticles. This method could detect the target virus as low as 11.16 μg ml-1. Furthermore, it opens new opportunities for sensitive and colorimetric detection of viruses and proteins.

  3. 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.

  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. 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

  6. 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

  7. Bifunctional colorimetric oligonucleotide probe based on a G-quadruplex DNAzyme molecular beacon.

    PubMed

    Zhang, Libing; Zhu, Jinbo; Li, Tao; Wang, Erkang

    2011-12-01

    A label-free bifunctional colorimetric oligonucleotide probe for DNA and protein detection has been developed on the basis of a novel catalytic molecular beacon consisting of two hairpin structures and a split G-quadruplex DNAzyme in the middle. The two loops of this molecular beacon consist of thrombin aptamer sequence and the complementary sequence of target DNA, which are utilized to sense single-stranded DNA and thrombin. The G-quadruplex DNAzyme can effectively catalyze the H(2)O(2)-mediated oxidation of 3,3',5,5'-tetramethylbenzidine sulfate to generate colorimetric signal. Upon addition of the target, the DNA or protein combines with one loop of the hairpin structures, and meanwhile drives the middle G-quadruplex DNAzyme to dissociate. This results in a decrease of catalytic activity, enabling the separate analysis of DNA and thrombin.

  8. Digital analysis technique for uncertainty reduction in colorimetric arsenic detection method.

    PubMed

    Carro Perez, Magali E; Francisca, Franco M

    2013-01-01

    This article proposes an alternative to increase the reliability and reproducibility of a colorimetric method to measure arsenic (As) concentrations. The method of analysis developed incorporates a digital analysis technique to eliminate the operator dependence of results, and As concentrations are quantitatively determined from digital levels computed from photographs of the colorimetric reaction that emerges during the test. This technique allows the sensitivity of the detection to be increased at low concentration ranges, which is of fundamental importance for the detection of As given the current acceptable limit for drinking water. The results obtained show a very good correlation between As concentrations determined by means of analytical laboratory techniques and the method proposed in this research.

  9. Upconversion ratiometric fluorescence and colorimetric dual-readout assay for uric acid.

    PubMed

    Fang, Aijin; Wu, Qiongqiong; Lu, Qiujun; Chen, Hongyu; Li, Haitao; Liu, Meiling; Zhang, Youyu; Yao, Shouzhuo

    2016-12-15

    A new upconversion colorimetric and ratiometric fluorescence detection method for uric acid (UA) has been designed. Yb(3+), Er(3+) and Tm(3+) co-doped NaYF4 nanoparticles (UCNPs) was synthesized. The co-doped NaYF4 nanoparticles, emit upconversion fluorescence with four typical emission peaks centered at 490nm, 557nm, 670nm and 705nm under the 980nm near-infrared (NIR) irradiation. The ZnFe2O4 magnetic nanoparticles (MNPs) possessing excellent peroxidase-like activity was prepared and used to catalyze oxidation the coupling of N-ethyl-N-(3-sulfopropyl)-3-methylaniline sodium salt (TOPS) and 4-amino-antipyrine (4-AAP) in the presence of H2O2 to form purple products (compound 1) which has a characteristic absorption peak located at 550nm. The upconversion fluorescence at 557nm was quenched by the compound 1 while the upconversion emission at 705nm was essentially unchanged, the fluorescence ratio ((I557/I705)0/(I557/I705)) is positively proportional to UA concentration in existence of uricase. More importantly, colorimetric signal can be easily observed and applied to directly distinguish the concentration of UA by the naked eye. Under the optimized conditions, the linear range of colorimetric and ratiometric fluorescence sensing towards UA was 0.01-1mM, the detection limits were as low as 5.79μM and 2.86μM (S/N=3), respectively. The proposed method has been successfully applied to the analysis of UA in human serum. These results indicate that the colorimetric and ratiometric fluorescence dual-readout assay method has great potential for applications in physiological and pathological diagnosis. PMID:27471157

  10. 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.

  11. Colorimetric assessment of BCR-ABL1 transcripts in clinical samples via gold nanoprobes.

    PubMed

    Vinhas, Raquel; Correia, Cláudia; Ribeiro, Patricia; Lourenço, Alexandra; Botelho de Sousa, Aida; Fernandes, Alexandra R; Baptista, Pedro V

    2016-07-01

    Gold nanoparticles functionalized with thiolated oligonucleotides (Au-nanoprobes) have been used in a range of applications for the detection of bioanalytes of interest, from ions to proteins and DNA targets. These detection strategies are based on the unique optical properties of gold nanoparticles, in particular, the intense color that is subject to modulation by modification of the medium dieletric. Au-nanoprobes have been applied for the detection and characterization of specific DNA sequences of interest, namely pathogens and disease biomarkers. Nevertheless, despite its relevance, only a few reports exist on the detection of RNA targets. Among these strategies, the colorimetric detection of DNA has been proven to work for several different targets in controlled samples but demonstration in real clinical bioanalysis has been elusive. Here, we used a colorimetric method based on Au-nanoprobes for the direct detection of the e14a2 BCR-ABL fusion transcript in myeloid leukemia patient samples without the need for retro-transcription. Au-nanoprobes directly assessed total RNA from 38 clinical samples, and results were validated against reverse transcription-nested polymerase chain reaction (RT-nested PCR) and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The colorimetric Au-nanoprobe assay is a simple yet reliable strategy to scrutinize myeloid leukemia patients at diagnosis and evaluate progression, with obvious advantages in terms of time and cost, particularly in low- to medium-income countries where molecular screening is not routinely feasible. Graphical abstract Gold nanoprobe for colorimetric detection of BCR-ABL1 fusion transcripts originating from the Philadelphia chromosome. PMID:27225178

  12. 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.

  13. A benzobisimidazolium-based fluorescent and colorimetric chemosensor for CO2.

    PubMed

    Guo, Zhiqian; Song, Na Ri; Moon, Jong Hun; Kim, Myounwoo; Jun, Eun Jin; Choi, Jiyoung; Lee, Jin Yong; Bielawski, Christopher W; Sessler, Jonathan L; Yoon, Juyoung

    2012-10-31

    A new sensor for the fluorescent and colorimetric detection of CO(2) is described. The system utilizes fluoride to activate a tetrapropyl benzobisimidazolium salt and operates in the absence of an exogenous base. On the basis of spectroscopic and theoretical analyses, the mode of action of the present system is ascribed to the fluoride-induced formation of an N-heterocyclic carbene intermediate that reacts with CO(2) to form an imidazolium carboxylate.

  14. 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

  15. 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.

  16. 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

  17. 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

  18. 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

  19. 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

  20. Evaluation of an end-tidal portable ETCO2 colorimetric breath indicator (COLIBRI).

    PubMed

    Rabitsch, Werner; Nikolic, Ajsa; Schellongowski, Peter; Kofler, Julia; Kraft, Peter; Krenn, Claus G; Staudinger, Thomas; Locker, Gottfried J; Knöbl, Paul; Hofbauer, Roland; Frass, Michael

    2004-01-01

    Evaluation of tube position is important after in-hospital and prehospital emergency intubation. Colorimetric breath indicators are devices for immediate control of tube positioning by showing a color change according to end-tidal CO2 (ETCO2) concentrations. We hypothesized that colorimetric breath indicators can yield reliable results for confirmation of tube position. The aim of this study was to evaluate the effectiveness and safety of a new colorimetric breath indicator (Colibri, ICOR AB, Bromma, Sweden) in 147 patients during general anesthesia, in critically ill patients, during transport to in-hospital interventions, and in a study design after insertion of a second tube into the esophagus in long-term ventilated patients. The Colibri was attached between the respective airway and ventilatory tubing. Seventy-three patients were investigated during general anesthesia, 39 patients were observed during long-term ventilation with an average duration of 33 hours, in 15 patients during transport to an intervention for up to 4 hours, and in 20 long-term ventilated patients at the medical intensive-care unit after insertion of a second tube intentionally into the esophagus with the help of a laryngoscope. The Colibri worked well in all groups investigated and showed no false results in the group with tubes inserted into the trachea and esophagus. Data suggest that the Colibri might serve as a valuable tool for evaluating and controlling tube position. This device is independent of power supply or electronic equipment, portable, small, and immediately ready for use.