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.

Enzyme-Linked Immunosorbent Assay for Detection of Zearalenone in Corn, Wheat, and Pig Feed: Collaborative Study

1994 ◽  
Vol 77 (6) ◽  
pp. 1500-1508 ◽  
Author(s):  
Glenn A Bennett ◽  
Terry C Nelsen ◽  
Brjnton M Miller
Keyword(s):  
Immunosorbent Assay ◽  
Collaborative Study ◽  
Screening Method ◽  
The United States ◽  
False Positives ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
False Negatives ◽  
Relative Standard ◽  
Pig Feed

Abstract A direct competitive enzyme-linked immunosorbent assay (ELISA) screening method for zearalenone in corn, wheat, and feed at 500 ng/g was evaluated by 23 collaborators (22 laboratories) in an international collaborative study. Eighteen samples of spiked or naturally contaminated corn, wheat, and pig feed were prepared by the sponsoring laboratory and sent for testing with complete test kits to participating collaborators in Canada, Italy, Sweden, The Netherlands, and the United States. Test samples were extracted with methanolwater solution (70 + 30) by shaking on a wrist-action shaker for 3 min. A portion of the extract was mixed with an equal volume of zearalenone-enzyme conjugate, and the mixture was incubated with zearalenone-specific monoclonal antibodies coated onto microtiter wells. All test samples were assayed in duplicate. One of 52 (2%) blanks was reported positive. Thirty-nine of the 52 (75%) samples that were spiked at 500 ng/g were reported as positive. Forty-nine of the 51 (96%) samples with concentrations at or above 1000 ng/g were reported as positive. The overall incidence of false negatives was 6.0% and the incidence of false positives was 22.7% by the ELISA method. Only one (3.4%) false negative was reported for samples containing ≥800 ng/g. In the spectrophotometric method, 8 collaborators determined approximate levels of zearalenone in test samples from standard curves constructed from spiked extracts (0–3000 ng/g of each commodity tested). This method gave and overall incidence of false negatives of 5.7% and false positives of 17.8%. Average relative standard deviations, RSDr (repeatability) and RSDR (reproducibility), were 11.6 and 25.1% for spiked samples and 11.7 and 33.1% for naturally contaminated samples, respectively. Standard curves were constructed with each set of samples assayed. Comparison of absorbance values from these standard curves indicate the performance of reagents and antibody used in the assay. The ELISA method has been adopted first action by AOAC INTERNATIONAL as a screening method for zearalenone at ≥800 ng/g in corn, wheat, and pig feed.

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.

MYCOTOXINS IN BEANS OF PEANUTS GROWN IN TAJIKISTAN

2018 ◽  
Vol 1 (4) ◽  
pp. 74-77
Author(s):  
Sh. I. Razokov ◽  
◽  
D. M. Mirzoev ◽  
G. P. Kononenko ◽  
A. A. Burkin ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Immunosorbent Assay ◽  
Cyclopiazonic Acid ◽  
Practical Significance ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Systems ◽  
The Republic ◽  
Combined Contamination

The article presents the results of an extensive mycotoxicological examination of 11 samples of peanut beans grown in two regions of the Republic of Tajikistan. The determination of 16 mycotoxins was carried out by indirect competitive enzyme-linked immunosorbent assay using commercial and certified research test systems. It has been established that for peanut beans in this area, a combined contamination by a group of sanitary-significant mycotoxins, including diacetoxyscirpenol, alternariol, ochratoxin A, PR-toxin and cyclopiazonic acid, is characteristic. The prospects of further research and the practical significance of the results are discussed.

A highly selective colorimetric assay for the determination of creatinine in biological samples using gluconic acid capped silver nanoparticles after ionic liquid based dispersive liquid phase microextraction

2021 ◽  
pp. 1-8
Author(s):  
Susan Sadeghi ◽  
Mohadeseh Hosseinpour-Zaryabi
Keyword(s):  
Silver Nanoparticles ◽  
Ionic Liquid ◽  
Liquid Phase ◽  
Human Serum ◽  
Gluconic Acid ◽  
Ag Nps ◽  
Liquid Phase Microextraction ◽  
Relative Standard ◽  
Serum And Urine

A dispersive liquid-phase microextraction method combined with UV–vis spectrophotometry was utilized to highly selective determination of creatinine in human serum and urine samples. To overcome the interferences in complex matrices, creatinine reacted with 1,4-naphthoquinone-2- potassium sulfonate reagent to produce a red coloured product that could be extracted into a small volume of 1-hexyl-3-methylimidazolium hexafluorophosphate ([HMIM]PF6) ionic liquid solvent. To increase the sensitivity of the assay, gluconic acid capped silver nanoparticles (Ag NPs) were used. On addition of Ag NPs to the red coloured extracted product, the solution turned to blue accompanied with a red shift in wavelength around 620 nm that could be detected by the naked eye. The effective variables on the determination of creatinine such as concentration of the reagent, amount of formic and hydrochloric acids, type and volume of the extractant, and concentration of Ag NPs were investigated. Under the optimal conditions, the calibration plot was bimodal with linear ranges from 0.1 to 1.5 µg mL−1 and 1.5 to 105 µg mL−1 creatinine with a limit of detection 0.1 µg mL−1. The relative standard deviation for five measurements at 35 µg mL−1 concentration level was 3.8%. The newly developed assay was used for the determination of creatinine in human serum and urine specimens with satisfactory results.

scholarly journals Interlaboratory Evaluation of Two Enzyme-Linked Immunosorbent Assay Kits for the Determination of Crustacean Protein in Processed Foods

2008 ◽  
Vol 91 (1) ◽  
pp. 123-129 ◽  
Author(s):  
Shinobu Sakai ◽  
Rieko Matsuda ◽  
Reiko Adachi ◽  
Hiroshi Akiyama ◽  
Tamio Maitani ◽  
...  
Keyword(s):  
Standard Deviation ◽  
Relative Standard Deviation ◽  
Immunosorbent Assay ◽  
Soluble Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Processed Foods ◽  
Relative Standard ◽  
High Level

Abstract The labeling of foods containing material derived from crustaceans such as shrimp and crab is to become mandatory in Japan because of increases in the number of allergy patients. To ensure proper labeling, 2 novel sandwich enzyme-linked immunosorbent assay (ELISA) kits for the determination of crustacean protein in processed foods, the N kit (Nissui Pharmaceutical Co., Ltd, Ibaraki, Japan) and the M kit (Maruha Nichiro Holdings, Inc., Ibaraki, Japan), have been developed. Five types of model processed foods containing 10 and/or 11.9 g/g crustacean soluble protein were prepared for interlaboratory evaluation of the performance of these kits. The N kit displayed a relatively high level of reproducibility relative standard deviation (interlaboratory precision; 4.08.4 RSDR) and sufficient recovery (6586) for all the model processed foods. The M kit displayed sufficient reproducibility (17.620.5 RSDR) and a reasonably high level of recovery (82103). The repeatability relative standard deviation (RSDr) values regarding the detection of crustacean proteins in the 5 model foods were mostly &lt;5.1 RSDr for the N kit and 9.9 RSDr for the M kit. In conclusion, the results of this interlaboratory evaluation suggest that both these ELISA kits would be very useful for detecting crustacean protein in processed foods.

scholarly journals Determination of Ochratoxin A in Wine and Beer by Immunoaffinity Column Cleanup and Liquid Chromatographic Analysis with Fluorometric Detection: Collaborative Study

2001 ◽  
Vol 84 (6) ◽  
pp. 1818-1827 ◽  
Author(s):  
Angelo Visconti ◽  
Michelangelo Pascale ◽  
Gianluca Centonze ◽  
E Anklam ◽  
A M Betbeder ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Red Wine ◽  
Collaborative Study ◽  
White Wine ◽  
Fluorometric Detection ◽  
Immunoaffinity Column ◽  
Relative Standard ◽  
Standard Deviations ◽  
Liquid Chromatographic

Abstract The accuracy, repeatability, and reproducibility characteristics of a liquid chromatographic method for the determination of ochratoxin A (OTA) in white wine, red wine, and beer were established in a collaborative study involving 18 laboratories in 10 countries. Blind duplicates of blank, spiked, and naturally contaminated materials at levels ranging from ≤0.01 to 3.00 ng/mL were analyzed. Wine and beer samples were diluted with a solution containing polyethylene glycol and sodium hydrogen carbonate, and the diluted samples were filtered and cleaned up on an immunoaffinity column. OTA was eluted with methanol and quantified by reversed-phase liquid chromatography with fluorometric detection. Average recoveries from white wine, red wine, and beer ranged from 88.2 to 105.4% (at spiking levels ranging from 0.1 to 2.0 ng/mL), from 84.3 to 93.1% (at spiking levels ranging from 0.2 to 3.0 ng/mL), and from 87.0 to 95.0% (at spiking levels ranging from 0.2 to 1.5 ng/mL), respectively. Relative standard deviations for within-laboratory repeatability (RSDr) ranged from 6.6 to 10.8% for white wine, from 6.5 to 10.8% for red wine, and from 4.7 to 16.5% for beer. Relative standard deviations for between-laboratories reproducibility (RSDR) ranged from 13.1 to 15.9% for white wine, from 11.9 to 13.6% for red wine, and from 15.2 to 26.1% for beer. HORRAT values were ≤0.4 for the 3 matrixes.

scholarly journals Determination of Picogram Levels of Roxithromycin in Pharmaceutical, Human Serum, and Urine by Flow-Injection Chemiluminescence

2012 ◽  
Vol 2012 ◽  
pp. 1-5
Author(s):  
Jiangman Liu ◽  
Huan Yang ◽  
Yun Zhang ◽  
Min Wu ◽  
Haixiang Zhao ◽  
...  
Keyword(s):  
Human Serum ◽  
Flow Injection ◽  
Limit Of Detection ◽  
Inhibitory Effect ◽  
Injection System ◽  
Relative Standard ◽  
Flow Injection System ◽  
Flow Injection Chemiluminescence ◽  
Serum And Urine

A sensitive chemiluminescence (CL) method, based on the inhibitory effect of roxithromycin (ROX) on the CL reaction between luminol and dissolved oxygen in a flow-injection system, was first proposed for the determination of ROX at picogram levels. The decrement of CL intensity was linearly proportional to the logarithm of ROX concentrations ranging from 0.1 to 100 pg mL-1, giving the limit of detection (LOD) of 0.03 pg mL-1 (3σ). At a flow rate of 2.0 mL min-1, a complete analytical procedure including sampling and washing could be performed within 0.5 min, with relative standard deviations (RSDs) of less than 5.0% (n=5). The proposed procedure was applied successfully to the determination of ROX in pharmaceutical, human serum, and urine with the recoveries ranging from 90.0 to 110.0%.

Determination of Total Aflatoxin Levels in Peanut Butter by Enzyme-Linked Immunosorbent Assay: Collaborative Study

1992 ◽  
Vol 75 (4) ◽  
pp. 693-697 ◽  
Author(s):  
Alan L Patey ◽  
Matthew Sharman ◽  
John Gilbert
Keyword(s):  
Immunosorbent Assay ◽  
Aflatoxin B1 ◽  
Peanut Butter ◽  
Collaborative Study ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Total Aflatoxin ◽  
Aflatoxin Level ◽  
Relative Standard

Abstract Laboratories in Australia, Japan, Spain, the United Kingdom, and the United States participated in a collaborative study to evaluate a commercial enzyme- linked immunosorbent assay for the determination of total aflatoxin. Collaborators were sent 10 randomly numbered samples (5 blind duplicates) of roasted peanut butter. Two pairs were "blank" peanut butters to which aflatoxin B1, B2, G1, and G2 standards had been added. The other 3 pairs of peanut butters were 1 low aflatoxin level sample and 2 naturally contaminated samples. The assay is based on indirect competition. Test samples containing (free) aflatoxin, added to aflatoxin-coated microwells, compete for specific monoclonal rat anti-aflatoxin. As the concentration of aflatoxin in the test samples increases, the amount of rat antiaflatoxin binding to the aflatoxin attached to the well decreases. After a wash step to remove unbound material, the amount of rat anti-aflatoxin bound to the well is determined by its reaction with peroxidase conjugated rabbit anti-rat globulin. Bound peroxidase activity is then determined by the addition of a substrate, whose color development is inversely proportional to the aflatoxin concentration and is measured by absorbance. Coefficients of variation (CV) for total aflatoxin concentrations, for mean levels of 9,30, and 89µg/kg, were between 28 and 37% for the low level and 2 naturally contaminated samples, which contained mainly aflatoxin B1. CVs for the spiked samples were lower (24-25%) for mean levels of 11 and 20 µg/kg; recoveries were 84 and 89%, respectively. Ranges for relative standard deviations for repeatabilty and reproducibility were 9-30% and 25-37%, respectively. The method has been adopted first action by AOAC International.

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 Detection of Five Mycotoxins in Different Food Matrices in the Malaysian Market by Using Validated Liquid Chromatography Electrospray Ionization Triple Quadrupole Mass Spectrometry

Toxins ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 196 ◽  
Author(s):  
Ali Alsharif ◽  
Yeun-Mun Choo ◽  
Guan-Huat Tan
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Ochratoxin A ◽  
Wheat Flour ◽  
Food Samples ◽  
Food Contaminants ◽  
Relative Standard ◽  
Liquid Chromatography Tandem Mass ◽  
Food Matrices

Mycotoxins are common food contaminants which cause poisoning and severe health risks to humans and animals. The present study applied chemometric approach in liquid chromatography-tandem mass spectrometry (LC-MS/MS) optimization for simultaneous determination of mycotoxins, i.e., aflatoxins B1, B2, G1, and G2, and ochratoxin A. The validated quick, easy, cheap, effective, rugged, and safe (QuEChERS)-LC-MS/MS method was used to study the occurrence of mycotoxins in 120 food matrices. The recovery ranges from 81.94% to 101.67% with relative standard deviation (RSD) lesser than 11%. Through the developed method, aflatoxins were detected in raisin, pistachio, peanut, wheat flour, spice, and chili samples with concentration ranges from 0.45 to 16.93 µg/kg. Trace concentration of ochratoxin A was found in wheat flour and peanut samples which ranged from 1.2 to 3.53 µg/kg. Some of the tested food samples contained mycotoxins of above the European legal maximum limit.

Sign in / Sign up

Export Citation Format

Share Document