scholarly journals Interlaboratory collaboration to determine the performance of the Randox food diagnostics biochip array technology for the simultaneous quantitative detection of seven mycotoxins in feed

2021 ◽  
pp. 1-10
Author(s):  
L. Sibanda ◽  
K. McCallum ◽  
M. Plotan ◽  
S. Webb ◽  
B. Snodgras ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Collaborative Study ◽  
Pass Rate ◽  
Quantitative Detection ◽  
Z Score ◽  
Control Programs ◽  
Feed Control ◽  
Array Technology ◽  
Fit For Purpose ◽  
Score Range

An inter-laboratory collaborative study was performed to evaluate the performance of the Biochip Array Technology (BAT) Myco 7 method. The Myco 7 Array is a method which simultaneously and quantitatively detects 20 mycotoxins including aflatoxins B1, B2, G1 and G2, ochratoxin A, deoxynivalenol, zearalenone, fumonisin B1, B2 and B3 and T-2 and HT-2 toxin. The BAT Myco 7 method was collaboratively evaluated by nine government and private Association of American Feed Control Officials (AAFCO) laboratories. Samples were analysed in a proficiency testing round format. Seventeen blind samples were analysed on the same equipment using Myco 7 kits. 99% of the results fell within an acceptable Z-score range of -2|<Z<|+2. Deoxynivalenol had a 100% Z-score pass rate, while a 99% pass was recorded for aflatoxins, zearalenone, ochratoxin A and fumonisins. T-2 toxin had a 97% Z-score pass rate. HorRat analysis for reproducibility used a range of 0.3<|HorRat|≤2. The target was met for deoxynivalenol, zearalenone, T-2 and HT-2 toxin, and aflatoxins B1, B2, G1 and G2 assays. Fumonisins and ochratoxin A assays had a 93% and 94% pass, respectively. The reproducibility co-efficiency of variation was between 16 and 20% meeting set criterion of <40% and is, therefore, fit-for-purpose for use in the AAFCO control programs for mycotoxins.

scholarly journals Dramatically Enhancing the Sensitivity of Immunoassay for Ochratoxin A Detection by Cascade-Amplifying Enzyme Loading

Toxins ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 781
Author(s):  
Zhuolin Song ◽  
Lin Feng ◽  
Yuankui Leng ◽  
Mingzhu Huang ◽  
Hao Fang ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Limit Of Detection ◽  
High Sensitivity ◽  
Cross Reaction ◽  
Molar Ratio ◽  
Quantitative Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Enzyme Loading ◽  
M13 Bacteriophage ◽  
Mimic Peptide

Enzyme-linked immunosorbent assay (ELISA) is widely used in the routine screening of mycotoxin contamination in various agricultural and food products. Herein, a cascade-amplifying system was introduced to dramatically promote the sensitivity of an immunoassay for ochratoxin A (OTA) detection. Specifically, a biotinylated M13 bacteriophage was introduced as a biofunctional competing antigen, in which a seven-peptide OTA mimotope fused on the p3 protein of M13 was used to specifically recognize an anti-OTA monoclonal antibody, and the biotin molecules modified on capsid p8 proteins were used in loading numerous streptavidin-labeled polymeric horseradish peroxidases (HRPs). Owing to the abundance of biotinylated p8 proteins in M13 and the high molar ratio between HRP and streptavidin in streptavidin-polyHRP, the loading amount of HRP enzymes on the M13 bacteriophage were greatly boosted. Hence, the proposed method exhibited high sensitivity, with a limit of detection of 2.0 pg/mL for OTA detection, which was 250-fold lower than that of conventional ELISA. In addition, the proposed method showed a slight cross-reaction of 2.3% to OTB, a negligible cross-reaction for other common mycotoxins, and an acceptable accuracy for OTA quantitative detection in real corn samples. The practicability of the method was further confirmed with a traditional HRP-based ELISA method. In conclusion, the biotinylated bacteriophage and polyHRP structure showed potential as a cascade-amplifying enzyme loading system for ultra-trace OTA detemination, and its application can be extended to the detection of other analytes by altering specific mimic peptide sequences.

Collaborative Study of a Method for the Determination of Ochratoxin A in Green Coffee

1975 ◽  
Vol 58 (2) ◽  
pp. 258-262 ◽  
Author(s):  
Colette P Levi
Keyword(s):  
Thin Layer ◽  
Ochratoxin A ◽  
Collaborative Study ◽  
Green Coffee ◽  
Average Recovery ◽  
Semiquantitative Determination ◽  
Chromatographic Methods ◽  
Green Coffee Beans ◽  
Coffee Beans

Abstract A method for the semiquantitative determination of ochratoxin A in green coffee has been studied collaboratively by 11 laboratories. The average recovery for the 7 samples spiked at 3 levels of ochratoxin A was 69.1%, ranging from 60.5 to 85.6%. This is comparable to other visual thin layer chromatographic methods of mycotoxin detection. The method has been adopted as official first action for the determination of ochratoxin A in green coffee beans.

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 Antibody Microarray Immunoassay for Simultaneous Quantification of Multiple Mycotoxins in Corn Samples

Toxins ◽  
2018 ◽  
Vol 10 (10) ◽  
pp. 415 ◽  
Author(s):  
Xian Zhang ◽  
Zuohuan Wang ◽  
Yun Fang ◽  
Renjie Sun ◽  
Tong Cao ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Aflatoxin B1 ◽  
Goodness Of Fit ◽  
Limit Of Detection ◽  
Simultaneous Detection ◽  
Fumonisin B1 ◽  
Quantitative Detection ◽  
Antibody Microarray ◽  
Test Kit ◽  
Microarray Immunoassay

We developed and tested a prototype of an antibody microarray immunoassay for simultaneous quantitative detection of four typical mycotoxins (aflatoxin B1, ochratoxin A, zearalenone, and fumonisin B1) in corn samples. The test kit consisted of a nitrocellulose membrane layered with immobilized monoclonal antibodies against mycotoxins. During the assay, the mycotoxin-protein conjugates were biotinylated. The signal detection was enhanced by a combination of the biotin-streptavidin system and enhanced chemiluminescence (ECL). This improved the sensitivity of the assay. Under the optimized conditions, four calibration curves with goodness of fit (R2 > 0.98) were plotted. The results showed that the detection limits for aflatoxin B1, ochratoxin A, zearalenone, and fumonisin B1 were 0.21, 0.19, 0.09, and 0.24 ng/mL, with detection ranges of 0.47–55.69, 0.48–127.11, 0.22–31.36, and 0.56–92.57 ng/mL, respectively. The limit of detection (LOD) of this antibody microarray for aflatoxin B1, ochratoxin A, zearalenone, and fumonisin B1 in corn was 5.25, 4.75, 2.25, and 6 μg/kg, respectively. The recovery rates from the spiked samples were between 79.2% and 113.4%, with coefficient of variation <10%. The results of the analysis of commercial samples for mycotoxins using this new assay and the liquid chromatography-tandem mass spectrometry (LC-MS/MS) were comparable and in good agreement. This assay could also be modified for the simultaneous detection of other multiple mycotoxins, as well as low-weight analytes, hazardous to human health.

scholarly journals Combined Phenyl Silane and Immunoaffinity Column Cleanup with Liquid Chromatography for Determination of Ochratoxin A in Roasted Coffee: Collaborative Study

2001 ◽  
Vol 84 (2) ◽  
pp. 444-450 ◽  
Author(s):  
A Catherine Entwisle ◽  
Alison C Williams ◽  
Peter J Mann ◽  
Joanne Russell ◽  
Philip T Slack ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Standard Deviation ◽  
Ochratoxin A ◽  
Relative Standard Deviation ◽  
Collaborative Study ◽  
Immunoaffinity Column ◽  
Roasted Coffee ◽  
Test Portion ◽  
Low Level ◽  
Relative Standard

Abstract A collaborative study was conducted to evaluate a liquid chromatography (LC) method for ochratoxin A using sequential phenyl silane and immunoaffinity column cleanup. The method was tested at 3 different levels of ochratoxin A in roasted coffee, which spanned the range of possible future European regulatory limits. The test portion was extracted with methanol and sodium bicarbonate by shaking for 30 min. The extract was filtered, centrifuged, and then cleaned up on a phenyl silane column before being eluted from the washed column with methanol–water. The eluate was diluted with phosphate-buffered saline (PBS) and applied to an ochratoxin A immunoaffinity column, which was washed with water. The ochratoxin A was eluted with methanol, the solvent was evaporated, and the residue was redissolved in injection solvent. After injection of this solution onto a reversed-phase LC apparatus, ochratoxin A was measured by fluorescence detection. Eight laboratory samples of low-level naturally contaminated roasted coffee and 2 laboratory samples of blank coffee (&lt; 0.2 ng/g ochratoxin A at the signal-to-noise ratio of 3:1), along with ampules of ochratoxin A calibrant and spiking solutions, were sent to 15 laboratories in 13 different European countries. Test portions of the laboratory samples were spiked at levels of 4 ng/g ochratoxin A, and recoveries ranged from 65 to 97%. Based on results for spiked blank material (blind duplicates) and naturally contaminated material (blind duplicates at 3 levels), the relative standard deviation for repeatability (RSDr) ranged from 2 to 22% and the relative standard deviation for reproducibility (RSDR) ranged from 14 to 26%. The method showed acceptable within- and between-laboratory precision, as evidenced by HORRAT values, at the low level of determination for ochratoxin A in roasted coffee.

Analysis of Ochratoxins A and B and Their Esters in Barley, Using Partition and Thin Layer Chromatography. II. Collaborative Study

1973 ◽  
Vol 56 (4) ◽  
pp. 822-826
Author(s):  
Stanley Nesheim
Keyword(s):  
Thin Layer ◽  
Ochratoxin A ◽  
Coefficient Of Variation ◽  
Collaborative Study ◽  
Ethyl Esters ◽  
The Other ◽  
Average Recovery ◽  
Confirmation Test ◽  
Two Samples ◽  
Layer Chromatography

Abstract To test the method of Nesheim et al., 6 samples wert; analyzed in 13 laboratories. The samples encompassed a blank and 5 samples containing one or more ochratoxins in the range 50–200 μg/kg. Two samples were spiked with the 4 ochratoxin standards and 3 were spiked with barley naturally contaminated with ochratoxin A. The confirmation of identity of ochratoxins A and B by preparation of their ethyl ester derivatives was also tested. The average recovery of standard ochratoxin A was 112% at levels of 45 and 90 μg/kg, with a 27.1% coefficient of variation calculated from analysis of variance, one analyst, one replicate. Similar satisfactory results were obtained for the ethyl esters of A and B at a level of 120 μg/kg. The results were unsatisfactory for ochratoxin B and for the esters of A and B at the 60 μg/kg level. The chemical confirmation test was satisfactory for both ochratoxins A and B. The method, including chemical confirmation, has been adopted as official first action as quantitative for ochratoxin A and qualitative for the other toxins.

Design and Synthesis of Target-Responsive Aptamer-Cross-linked Hydrogel for Visual Quantitative Detection of Ochratoxin A

2015 ◽  
Vol 7 (12) ◽  
pp. 6982-6990 ◽  
Author(s):  
Rudi Liu ◽  
Yishun Huang ◽  
Yanli Ma ◽  
Shasha Jia ◽  
Mingxuan Gao ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Quantitative Detection ◽  
Design And Synthesis

Selection of phage-displayed minotopes of ochratoxin A and its detection in cereal by ELISA

2015 ◽  
Vol 7 (5) ◽  
pp. 1849-1854 ◽  
Author(s):  
Yao Wang ◽  
Xiaofei Hu ◽  
Yafeng Pei ◽  
Yaning Sun ◽  
Fangyu Wang ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Competitive Elisa ◽  
Quantitative Detection ◽  
Random Peptide ◽  
Peptide Display ◽  
Selection Of ◽  
Peptide Display Library

A competitive ELISA (cELISA) for the quantitative detection of ochratoxin A (OTA) was developed that uses a clone selected from a phage random peptide display library.

scholarly journals Application of Immunoaffinity Columns to Mycotoxin Analysis

1997 ◽  
Vol 80 (5) ◽  
pp. 941-950 ◽  
Author(s):  
Peter M Scott ◽  
Mary W Trucksess
Keyword(s):  
Ochratoxin A ◽  
Solid Phase ◽  
Biological Fluids ◽  
Peanut Butter ◽  
Collaborative Study ◽  
Automated System ◽  
Official Method ◽  
Buffer Solution ◽  
Test Kit ◽  
Immunoaffinity Columns

Abstract Immunoaffinity columns (lACs) are widely used for cleanup and isolation of mycotoxins extracted from foods and biological fluids, particularly afla-toxins, ochratoxin A, and fumonisinsẳ The columns are prepared by binding antibodies specific for a given mycotoxin to a specially activated solid-phase support and packing the support suspended in aqueous buffer solution into a cartridge. The mycotoxin in the extract or fluid binds to the antibody, impurities are removed with water or aqueous solution, and then the mycotoxin is desorbed with a miscible solvent such as methanol. Further separation can be performed with lAC, followed by liquid chromatographic (LC) quantitation, either off-line or on-line in an automated system, or by fluorometryằ lACs have been used by laboratories that developed the antibodies but are also available commercially for aflatoxins, ochratoxin A, fumonis-ins, zearalenone, and deoxynivalenolễ Among commercial lACs, Aflatest P is used as the cleanup step in an LC method and in a solution fluorometry method for corn, peanuts, and peanut butter that was adopted as an AOAC INTERNATIONAL Official Method after evaluation by an international collaborative study. As part of a fluorometer-based test kit, aflatest P was further certified by the AOAC Research Institute to measure total aflatoxins in 10 grains and grain products. lACs can concentrate the analyte from a large amount of sample, allowing detection limits at low parts-per-trillion levels in some cases (e.g., for aflatoxin Mi and ochratoxin A in liquid food matrixes). Regeneration of lACs for reuse in aflatoxin, ochratoxin A, fumon-isin, and zearalenone analyses has been investigated.

Determination of Ochratoxin A in Capsicum spp. (Paprika and Chili) by Immunoaffinity Column Cleanup and Liquid Chromatography: Collaborative Study

2014 ◽  
Vol 97 (3) ◽  
pp. 876-883 ◽  
Author(s):  
Zoltan Kunsagi ◽  
Joerg Stroka ◽  
E Abilleira ◽  
P Bastijns ◽  
M Berger ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Collaborative Study ◽  
Immunoaffinity Column ◽  
Fluorescence Detector ◽  
International Union ◽  
Eu Member States ◽  
Rp Hplc ◽  
Official Control ◽  
Capsicum Spp

Abstract A method validation study for the determination of ochratoxin A in Capsicum spp. (paprika and chili) was conducted according to the International Union of Pure and Applied Chemistry harmonized protocol. The method is based on the extraction of samples with aqueous methanol, followed by an immunoaffinity column cleanup. The determination is carried out by RP-HPLC coupled with a fluorescence detector. The study involved 21 participants representing a cross-section of research, private, and official control laboratories from 14 European Union (EU) Member States and Singapore. Mean recoveries reported ranged from 83.7 to 87.5%. The RSD for repeatability (RSDr) ranged from 1.7 to 14.3%. The RSD for reproducibility (RSDR) ranged from 9.1 to 27.5%, reflecting HorRat values from 0.4 to 1.3 according to the Horwitz function modified by Thompson. The correction for recovery of results from naturally contaminated samples further improved the reproducibility of the method. The method showed acceptable within-laboratory and between-laboratory precision for each matrix, and it conforms to requirements set by current EU legislation.

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