scholarly journals Simultaneous and fast determination of antibiotics as nitrofuran metabolites in fish and shrimp muscle using Enzyme-Linked Immunosorbent Assay (ELISA)

2020 ◽  
Vol 32 (1) ◽  
pp. 193-198
Author(s):  
MD. MEZANUR RAHMAN ◽  
MD. MAHMUDUL HASAN RONY ◽  
K.M. GOLZAR HOSSAIN ◽  
MD. GOLAM MOSTOFA ◽  
SALMA BEGUM ◽  
...  
Keyword(s):  
Screening Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
Performance Requirements ◽  
Accuracy And Precision ◽  
Liquid Chromatography Tandem Mass ◽  
Quantitative Screening ◽  
Method Accuracy ◽  
Nitrofuran Metabolites

A simultaneous and fast analytical method was developed for the identification of fournitrofuran metabolites from the fish and shrimp samples. Homogenized samples were hydrolyzedand derivatized with 4-nitrobenzaldehyde. Subsequently, extracted with ethyl acetate, evaporatedto dryness and the residue was re-dissolved in Hexane. Commercial enzyme-linkedimmunosorbent assay (ELISA) method was applied for the analysis of nitrofuran metabolites. Themethod was validated in shrimp and fish matrix according to the criteria defined in CommissionDecision 2002/657/EC for qualitative screening method following the guidelines set by thecommunity reference laboratories residues (CRLs) 2010. Characteristic’s parameters as detectioncapability (CC?), specificity/selectivity, stability, recovery and precision were determined.Detection capability (CC?) for nitrofuran metabolites (AMOZ, AOZ, AHD and SEM) in Fishand shrimp matrix was in the range of 0.5- 0.75?g/kg, which were less than the MinimumRequired Performance Limit (MRPL) of 1?g/kg set by European Union. The proposed method issuitable for semi-quantitative screening analysis of antibiotics in the fish and shrimp muscle inconformity with the current EU performance requirements before exporting to EU and othercountries. Results from analysis of unknown samples by the developed ELISA method werecomparable to those obtained by a liquid chromatography-tandem mass spectrometry (LCMS/MS) method. Accuracy and precision of the method had also been checked through participation of International Proficiency Testing (PT) having a very satisfactory performance score.

Determination of ochratoxin A in serum and urine of pigs

2012 ◽  
Vol 5 (4) ◽  
pp. 351-356 ◽  
Author(s):  
N. Perši ◽  
J. Pleadin ◽  
A. Vulić ◽  
I. Kmetić ◽  
B. Šimić
Keyword(s):  
Ochratoxin A ◽  
Method Validation ◽  
Screening Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
Relative Standard ◽  
Experimental Group ◽  
Simple Screening ◽  
Serum And Urine

The objective of the study was to determine ochratoxin A (OTA) concentrations in serum and urine of pigs during 30-day OTA treatment. OTA was administered orally to the experimental group (n=5) at a dose of 0.78 mg per animal per day, whereas control animals (n=5) were left untreated. OTA concentrations were determined using a validated enzyme-linked immunosorbent assay (ELISA). Method validation resulted in mean recoveries of 93-101% for serum and 98-106% for urine, with acceptable mean inter- and intraday relative standard deviations (<8% for urine and <7% for serum). The ELISA method can be effectively used as a simple screening method to determine OTA exposure in pigs during fattening. The maximum mean OTA concentration in serum was recorded on day 22 (8.75±2.93 ng/ml) and in urine on day 20 (43.56±35.76 ng/ml), indicating significant differences in OTA concentrations between these two matrices.

scholarly journals Simplified and Rapid Determination of Primaquine and 5,6-Orthoquinone Primaquine by UHPLC-MS/MS: Its Application to a Pharmacokinetic Study

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4357
Author(s):  
Waritda Pookmanee ◽  
Siriwan Thongthip ◽  
Jeeranut Tankanitlert ◽  
Mathirut Mungthin ◽  
Chonlaphat Sukasem ◽  
...  
Keyword(s):  
Pharmacokinetic Study ◽  
High Performance ◽  
Rapid Determination ◽  
Active Metabolites ◽  
Chromatography Tandem Mass Spectrometry ◽  
Accuracy And Precision ◽  
The Matrix ◽  
Liquid Chromatography Tandem Mass ◽  
Isocratic Mode

The method for the determination of primaquine (PQ) and 5,6-orthoquinone primaquine (5,6-PQ), the representative marker for PQ active metabolites, via CYP2D6 in human plasma and urine has been validated. All samples were extracted using acetonitrile for protein precipitation and analyzed using the ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) system. Chromatography separation was carried out using a Hypersil GOLDTM aQ C18 column (100 × 2.1 mm, particle size 1.9 μm) with a C18 guard column (4 × 3 mm) flowed with an isocratic mode of methanol, water, and acetonitrile in an optimal ratio at 0.4 mL/min. The retention times of 5,6-PQ and PQ in plasma and urine were 0.8 and 1.6 min, respectively. The method was validated according to the guideline. The linearity of the analytes was in the range of 25–1500 ng/mL. The matrix effect of PQ and 5,6-PQ ranged from 100% to 116% and from 87% to 104% for plasma, and from 87% to 89% and from 86% to 87% for urine, respectively. The recovery of PQ and 5,6-PQ ranged from 78% to 95% and form 80% to 98% for plasma, and from 102% to from 112% to 97% to 109% for urine, respectively. The accuracy and precision of PQ and 5,6-PQ in plasma and urine were within the acceptance criteria. The samples should be kept in the freezer (−80 °C) and analyzed within 7 days due to the metabolite stability. This validated UHPLC-MS/MS method was beneficial for a pharmacokinetic study in subjects receiving PQ.

Comparison of Two Immunochemical Methods with Thin-Layer Chromatographic Methods for Determination of Aflatoxins

1990 ◽  
Vol 73 (3) ◽  
pp. 425-428
Author(s):  
Mary W Trucksess ◽  
Kathryn Young ◽  
Kevin F Donahue1 ◽  
Deborah K Morris ◽  
Ernie Lewis
Keyword(s):  
Thin Layer ◽  
Correct Response ◽  
Peanut Butter ◽  
Screening Method ◽  
Elisa Test ◽  
Poultry Feed ◽  
Enzyme Linked Immunosorbent Assay ◽  
Affinity Column ◽  
Negative Results

Abstract Three different methods were compared for the determination of total flatoxins in corn and peanuts naturally contaminated with aflatoxins and In corn, peanuts, cottonseed, peanut butter, and poultry feed spiked with aflatoxins Bi, B2, and d . The 3 methods were an enzyme-linked Immunosorbent assay (ELISA) screening test; a monoclonal antibody-affinity column-solld-phase separation method; and the AOAC official thin-layer chromatography (TLC) methods for all except poultry feed, for which Shannon's TLC method for mixed feed was used. The ELISA test is designed to provide only positive results for total aflatoxins at &gt;20 ng/g or negative results at &lt;20 ng/g. The affinity column separation Is coupled with either brominatlon solution fluorometry to estimate total aflatoxins or liquid chromatography (LC) to quantltate Individual aflatoxins. Fluorodensitometry was used to determine aflatoxins In commodities analyzed by the TLC methods. The LC and TLC results were In good agreement for all the analyses. The results for the affinity column using brominatlon solution fluorometry were similar except those for cottonseed, which were about 60% higher. The ELISA screening method correctly Identified naturally contaminated corn and peanut positive samples. No false positives were found for controls. The correct response for spiked corn, raw peanuts, peanut butter, and cottonseed at &gt;20 ng aflatoxlns/g was about 90%. The correct response for spiked poultry feed at &gt;20 ng aflatoxlns/ g was about 50%.

scholarly journals Determination of Matrine in Rat Plasma after Oral Administration of Novel Korean Herbal Medicine KIOM-MA128 and Application of PK

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Hyun-moon Back ◽  
Byungjeong Song ◽  
Jung-woo Chae ◽  
Hwi-yeol Yun ◽  
Jin Yeul Ma ◽  
...  
Keyword(s):  
Herbal Medicine ◽  
Oral Administration ◽  
High Performance ◽  
Rat Plasma ◽  
Pharmacokinetic Studies ◽  
Chemical Marker ◽  
Chromatography Tandem Mass Spectrometry ◽  
Accuracy And Precision ◽  
Liquid Chromatography Tandem Mass

KIOM-MA128 is a novel Korean herbal medicine with antiatopic, anti-inflammatory, and antiasthmatic effects. Matrine is thought to be a potential chemical marker of KIOM-MA128, but pharmacokinetic studies on KIOM-MA128 had not been performed. This study describes a simple and rapid method using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) to determine the concentration of matrine in rats plasma after administration of KIOM-MA128. The isocratic mobile phase consisted of methanol and distilled water, and the flow rate was 0.15 mL/min. The accuracy and precision of the assay, as well as stability tests, were performed in accordance with FDA regulations for the validation of bioanalytical methods. The half-life andTmaxof matrine after administration of KIOM-MA128 were 4.29 ± 2.20 h and 1.8 ± 1.23 h, respectively.CmaxandAUCinfof matrine after administration of KIOM-MA128 at 4 g/kg and 8 g/kg were 595.10 ± 182.91 ng/mL, 5336.77 ± 1503.84 ng/mL·h and 850.46 ± 120 ng/mL, 9583.10 ± 888.92 ng/mL·h, respectively. The validated method was successfully applied to a pharmacokinetic study in rats after oral administration of KIOM-MA128.

A Rapid Wire-Loop Method for Determination of Lead in Paint by Atomic Absorption Spectrometry

1976 ◽  
Vol 30 (1) ◽  
pp. 56-63 ◽  
Author(s):  
M. Kubota ◽  
D. W. Golightly ◽  
R. Mavrodineanu
Keyword(s):  
Screening Method ◽  
Absorption Spectrometry ◽  
Acidic Aqueous Solution ◽  
Wire Loop ◽  
Loop Method ◽  
Accuracy And Precision ◽  
Small Resistance ◽  
Physical And Chemical ◽  
Liquid Paint

A simple, rapid method is described for the determination of lead in paint. Lead is extracted from paint chips, or liquid paint, into an acidic aqueous solution. Ten microliters of sample solution is transferred onto a platinum-rhodium wire loop, and the solvent is evaporated by placing the loop in a small resistance heater. The loop then is introduced into a premixed acetylene-air flame, and lead absorption at 283.3 nm is measured. Physical and chemical parameters affecting the accuracy and precision of the method are discussed. Accurate determinations of lead are possible for concentrations ranging from 0.1 to 1 µg/10 µl of solution. The wire loop method is adaptable to a simple screening method for analysis of paints in mobile or on-site laboratories.

scholarly journals High-Throughput Screening Method of Inhibitors that Block the Interaction between 2 Helical Regions of HIV-1 gp41

2005 ◽  
Vol 10 (1) ◽  
pp. 13-19 ◽  
Author(s):  
Bong-Suk Jin ◽  
Won-Kyu Lee ◽  
Kwangseog Ahn ◽  
Myung Kyu Lee ◽  
Yeon Gyu Yu
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Screening Method ◽  
Assay Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Compound Library ◽  
Elisa Method ◽  
Helical Bundle ◽  
Fusion Inhibitors ◽  
Hiv 1

The HIV-1 envelope glycoprotein transmembrane subunit, gp41, mediates the fusion of viral and target cell membranes. The 2 helical regions in the ectodomain of gp41, the N-helix and the C-helix, form a helical bundle complex that has been suggested as a fusion-active conformation. Previously, an enzyme-linked immunosorbent assay (ELISA) method had been established to measure the interaction of 2 helical regions of gp41. In this study, the ELISA method was modified to apply high-throughput screening (HTS) of an organic compound library. A few compounds had been identified to prevent the interaction between 2 helical regions of gp41, and they were further shown to inhibit the gp41-mediated viral infection. In addition, they specifically quenched the fluorescence of tryptophan in the N-helix region, indicating that these compounds bound to the N-helix rather than the C-helix of gp41. These results suggested that this assay method targeting gp41 could be used for HTS of HIV fusion inhibitors. ( Journal of Biomolecular Screening 2005:13-19)

Development and Single-Laboratory Validation of an Improved Method for the Determination of Cyclamate in Foods Using Liquid Chromatography/Tandem Mass Spectrometry

2014 ◽  
Vol 97 (6) ◽  
pp. 1651-1655 ◽  
Author(s):  
Romina Shah ◽  
Lowri S De Jager ◽  
Timothy H Begley
Keyword(s):  
Internal Standard ◽  
Pomegranate Juice ◽  
Spectral Determination ◽  
Precision Data ◽  
Mass Spectral ◽  
Minimal Sample ◽  
Accuracy And Precision ◽  
Liquid Chromatography Tandem Mass ◽  
Potential Matrix

Abstract A fast and reliable LC-MS/MS method for the determination of cyclamate in a variety of food matrices was developed and validated. This method provides both quantitation and qualitative mass spectral determination important for analysis of regulatory samples. Utilization of a cyclamate-d11 internal standard corrects for potential matrix interferences during sample injection and allows minimal sample preparation. Seventeen commercially available food products were fortified at 250 μg/mL and tested as part of the method validation. Recoveries ranged from 72 to 110%, with RSDs ranging from 3 to 15%. The linear range spanned 0.010–1.00 μg/mL. LODs were 0.1 and 0.6 ng/mL, determined in pomegranate juice and dried fig, respectively. LOQs were 0.3 and 1.6 ng/mL, which are significantly lower than needed to measure cyclamate when used as a food additive. The interday and intraday accuracy and precision data are presented. This method was validated for analysis of a variety of commonly adulterated products, including drinks, dried fruits, jams, and hard candies.

scholarly journals Validation of the method of quantitative determination of florphenicol in muscle samples by the method of immuno-enzyme analysis

2020 ◽  
pp. 40-45
Author(s):  
M. V. Kostyuk ◽  
◽  
K. S. Myagka ◽  
G. S. Kochetova ◽  
◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Screening Method ◽  
Enzyme Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Laboratory Diagnostics ◽  
Screening Methods ◽  
Detection Ability ◽  
Wide Range ◽  
Control Samples

Introduction. Amphenicols are a group of chemical compounds with antibacterial activity, including chloramphenicol (HAF), thiamphenicol (TAF), florfenicol (FF) and their derivatives. Florfenicol (FF) is a synthetic antimicrobial agent with a broad spectrum of action and is one of the most commonly used drugs in poultry, and was developed specifically for veterinary medicine. Given the wide range of activity of florfenicol, the high therapeutic effect combined with low toxicity makes it important for use in animal husbandry. The known method of enzyme-linked immunosorbent assay (ELISA) differs favorably from other screening methods by high sensitivity, specificity, simplicity and speed of performance, availability and stability of reagents, the ability to computer processing of measurement results and automation of test steps, which provides high test efficiency. The aim of the work. To validate the method of enzyme-linked immunosorbent assay to determine the residues of florfenicol in the samples of months of different species of animals (cattle, pigs, chickens, geese, turkeys, rabbits) and fish. Materials and methods. The research was conducted on the basis of the State Research Institute for Laboratory Diagnostics and Veterinary Sanitary Expertise. The material for the study was a solution of florfenicol with a concentration of 1 mg/liter. Results of research and discussion. It was found that the highest value (highest response) for control samples is 0.24 μg/kg and the lowest value for enriched samples (lowest response) - 3.62 μg/kg. According to the obtained results, none of the answers for the enriched samples coincides with the range of answers for the control samples. It follows that the detection ability (CCβ) for this screening method reduces or decreases 5.0 μg/kg The cut-off rate of this test is 3.62 μg/kg. Conclusions and prospects for further research. Validation characteristics have been established for the determination of florfenicol residues in muscle samples, such as: detection ability (CCβ) is 5.0 μg/kg, cut-off level is 3.62 μg/kg. The lowest content of florfenicol that can be determined is 0.2 μg/kg. The percentage of return for enriched samples of both groups is 93 %, which corresponds to the specified production.

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