Comparison of two Analytical Methods (ELISA and LC-MS/MS) for Determination of Aflatoxin B1 in Corn and Aflatoxin M1 in Milk

Background: Mycotoxins are the secondary metabolites of molds and have adverse effects on humans, animals, and crops, resulting in illnesses and economic losses. Aflatoxin M1 (AFM1) is a hepatocarcinogen found in the milk from animals that have consumed feeds contaminated with aflatoxin B1 (AFB1). Milk is a highly nutritious food and is a source of necessary macro- and micro-nutrients for the growth, development and maintenance of human health. Methods: The presence of AFM1 was investigated in 70 samples of imported pasteurized and powdered milk products available to the Iranian consumers. The level of AFM1 was determined by HPLC method equipped with immunoaffinity cleanup. Results: The results showed that 32% of the analyzed samples were positive for AFM1 at 0.05-3.31 μg/kg. Also, 16% of analyzed samples were positive for AFM1at concentrations higher than the limit permitted by the Iranian standards. Conclusion: The detection of AFM1 contamination in the analyzed samples indicates the importance of the health of animal feeds. Thus, monitoring the imported feed materials, especially those arriving at Iranian borders is crucial in the prevention of AFM1 and AFB1 contaminations spreading across the domestic market.

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An Impedance Based Electrochemical Immunosensor for Aflatoxin B1 Monitoring in Pistachio Matrices

Chemosensors ◽

10.3390/chemosensors8040121 ◽

2020 ◽

Vol 8 (4) ◽

pp. 121

Author(s):

Michail D. Kaminiaris ◽

Sophie Mavrikou ◽

Maria Georgiadou ◽

Georgia Paivana ◽

Dimitrios I. Tsitsigiannis ◽

...

Keyword(s):

Aflatoxin B1 ◽

Limit Of Detection ◽

Electrochemical Immunosensor ◽

Aflatoxin M1 ◽

Biomolecular Recognition ◽

Effective Prevention ◽

Upper Limits ◽

Ethylcarbodiimide Hydrochloride ◽

Food And Feed

Aflatoxins are highly toxic fungal secondary metabolites that often contaminate food and feed commodities. An electrochemical immunosensor for the determination of aflatoxin B1 (AFB1) was fabricated by immobilizing monoclonal AFB1 antibodies onto a screen-printed gold electrode that was modified with carbo-methyldextran by N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide cross-linking. An electrochemical interfacial modelling of biomolecular recognition was suggested and reasonably interpreted. Impedance technology was employed for the quantitative determination of AFB1. The limit of detection concentration of AFB1 for standard solutions and spiked pistachio samples was 0.5 ng/mL and 1 ng/mL, respectively. The immunosensor was able to successfully determine AFB1 concentrations in the range of 4.56–50.86 ng/mL in unknown pistachio samples. Comparative chromatographic analysis revealed that AFB1 concentrations that were higher than 345 ng/mL were not within the immunosensor’s upper limits of detection. Selectivity studies against Ochratoxin A and Aflatoxin M1 demonstrated that the proposed AFB1 immunosensor was able to differentiate between these other fungal mycotoxins. The novel electrochemical immunosensor approach has the potential for rapid sample screening in a portable, disposable format, thus contributing to the requirement for effective prevention and the control of aflatoxin B1 in pistachios.

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Determination of aflatoxins in food products by the ELISA method

Czech Journal of Food Sciences ◽

10.17221/6567-cjfs ◽

2013 ◽

Vol 19 (No. 1) ◽

pp. 8-12 ◽

Cited By ~ 8

Author(s):

J. Leszczyńska ◽

J. MasŁowska ◽

A. Owczarek ◽

U. Kucharska

Keyword(s):

Aflatoxin B1 ◽

Dairy Products ◽

Total Content ◽

Food Products ◽

Aflatoxin M1 ◽

Elisa Method ◽

Cereal Samples

To determine the total content of aflatoxins, aflatoxin B1 and aflatoxin M1 in food the ELISA method was used. Milk, dairy products and cereal samples were mainly investigated. A few samples were found to be contaminated with aflatoxins. A great usability of the ELISA method for aflatoxin determination in food was established. Selectivity and sensitivity of the method is reported.

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Extraction, Cleanup, and Quantitative Determination of Aflatoxins B1 and M1 in Beef Liver

Journal of AOAC INTERNATIONAL ◽

10.1093/jaoac/62.5.1080 ◽

1979 ◽

Vol 62 (5) ◽

pp. 1080-1082

Author(s):

Mary W Trucksess ◽

Leonard Stoloff

Keyword(s):

Citric Acid ◽

Thin Layer ◽

Quantitative Determination ◽

Aflatoxin B1 ◽

Aflatoxin M1 ◽

Animal Tissues ◽

Beef Liver ◽

Layer Chromatography ◽

Extracting Solvent

Abstract A method for the determination of aflatoxin Bx in eggs was applicable for aflatoxin B1 in liver, but ineffective for aflatoxin Mx in liver because of poor recovery of added aflatoxin and interferences in thin layer chromatography. The method was modified by the addition of citric acid to the extracting solvent and ammonium sulfate to the extract solution for removing protein. The elution system for silica gel column cleanup was also changed by substituting methanol for acetone, and adding a step for confirmation of aflatoxin M1 identity. The method has been used successfully for survey and research on aflatoxin residues in animal tissues.

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Determination of aflatoxin M1 in urine samples indicates frequent dietary exposure to aflatoxin B1 in the Bangladeshi population

International Journal of Hygiene and Environmental Health ◽

10.1016/j.ijheh.2016.11.002 ◽

2017 ◽

Vol 220 (2) ◽

pp. 271-281 ◽

Cited By ~ 18

Author(s):

Nurshad Ali ◽

Meinolf Blaszkewicz ◽

Khaled Hossain ◽

Gisela H. Degen

Keyword(s):

Aflatoxin B1 ◽

Dietary Exposure ◽

Urine Samples ◽

Aflatoxin M1 ◽

Bangladeshi Population

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DETERMINATION OF AFLATOXIN M1 LEVELS IN BOVINE FARMED MILK WITH SPECIAL REFERENCE TO THE LEVELS OF AFLATOXIN B1 IN THE ANIMAL FEED

Assiut Veterinary Medical Journal ◽

10.21608/avmj.2011.176919 ◽

2011 ◽

Vol 57 (131) ◽

pp. 1-11

Keyword(s):

Special Reference ◽

Aflatoxin B1 ◽

Animal Feed ◽

Aflatoxin M1

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Aflatoxin M in Perspective1

Journal of Food Protection ◽

10.4315/0362-028x-43.3.226 ◽

1980 ◽

Vol 43 (3) ◽

pp. 226-230 ◽

Cited By ~ 45

Author(s):

LEONARD STOLOFF

Keyword(s):

United States ◽

Drug Administration ◽

Aflatoxin B1 ◽

Peanut Meal ◽

The United States ◽

Aflatoxin M1 ◽

Milk Products ◽

Survey Results ◽

Commercial Milk

The steps are presented from the discovery of aflatoxin M1 in the milk of a cow fed “toxic” peanut meal to the determination of its relation to aflatoxin B1 and the relation of aflatoxin M1 to other animal metabolites of aflatoxin B1. Estimates from controlled studies are given of the level and amount of aflatoxin M1 to be expected in the milk of cows exposed to aflatoxin B1 in their feed, followed by numerous survey results that corroborate the conclusion from the experimental estimates: levels of aflatoxin B1 in uncontrolled dairy ration ingredients are sufficiently high to result in measurable aflatoxin M1 ( > 0.1 ng/ml) in commercial milk supplies. Because the Food and Drug Administration has been unable to prevent occurrence of aflatoxin M1 in milk in the United States by attempting to control the feed, an administrative guideline has been established at 0.5 ppb for aflatoxin M1 in fluid milk products; consideration is being given to related guidelines for products manufactured from milk.

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Ultra-high performance liquid chromatographic determination of aflatoxin M1 in urine

World Mycotoxin Journal ◽

10.3920/wmj2014.1761 ◽

2015 ◽

Vol 8 (4) ◽

pp. 405-413 ◽

Cited By ~ 3

Author(s):

S. Mohd Redzwan ◽

R. Jamaluddin ◽

A.M. Mohd Sokhini ◽

A.R. Nurul Aqilah ◽

A. Zuraini ◽

...

Keyword(s):

Biological Samples ◽

Analytical Methods ◽

High Performance ◽

Limit Of Detection ◽

Extraction Procedure ◽

Aflatoxin M1 ◽

Enzyme Linked Immunosorbent Assay ◽

Fluorescence Detector ◽

The Mean

The development of analytical methods to detect aflatoxin B1 (AFB1) in foodstuffs and its metabolites in human biological samples is useful for risk assessment. The latter methodology, i.e. the measurement of AFB1 biomarkers, has become important to assess human aflatoxin exposure. AFB1-lysine adduct, AFB1-DNA adduct and urinary aflatoxin M1 (AFM1) are some of the AFB1 biomarkers that can be measured by several analytical methods, such as enzyme-linked immunosorbent assay, radioimmunoassay, and high performance liquid chromatography (HPLC). HPLC coupled to a fluorescence detector is useful and preferable due to its high degree of sensitivity, but the analysis may take time and consume large amount of solvents. Therefore, the present study extrapolated the HPLC method to ultra-HPLC for the determination of urinary AFM1. After the extraction procedure with an immunoaffinity column, chromatographic separation was done using a high performance 1.8 μm microparticulate C18 column. The mean recovery from urine samples spiked with 0.5, 1.0 and 2.0 ng/ml AFM1 was 84.4±4.0%, with acceptable recovery values, interday (6.0±5.3%) and intraday (2.6±0.6%) coefficients of variation. The retention time was 5.7 min. This method was used to measure urinary AFM1 in 71 subjects, of which 13 had AFM1 levels above the limit of detection (0.018 ng/ml). The mean urinary AFM1 level of the positive samples was 18.8±28.6 pg/ml, ranging from 2.4 to 100.4 pg/ml. As this is one of the few studies investigating the occurrence of aflatoxin biomarkers in human biological samples in Malaysia, a study with a larger sample size is necessary to investigate the magnitude of aflatoxin exposure among the population.

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Reliable HPLC Determination of Aflatoxin M1 in Eggs

Journal of Analytical Methods in Chemistry ◽

10.1155/2013/817091 ◽

2013 ◽

Vol 2013 ◽

pp. 1-5 ◽

Cited By ~ 2

Author(s):

Mostafa M. H. Khalil ◽

Ahmed M. Gomaa ◽

Ahmed Salem Sebaei

Keyword(s):

Aflatoxin B1 ◽

High Performance ◽

Limit Of Detection ◽

Aflatoxin M1 ◽

Limit Of Quantification ◽

Chromatography Method ◽

Animal Products ◽

Liquid Chromatography Method ◽

Different Levels

Aflatoxin M1 is the foremost metabolite of aflatoxin B1 in humans and animals, which may be present in animal products from animals fed with aflatoxin B1 contaminated feed. In this study a high performance liquid chromatography method for determination of aflatoxin M1 in eggs was described. The egg samples were diluted with warmed water and the toxin was immunoextracted followed by fluorescence detection. The average recovery of aflatoxin M1 at the three different levels 0.05, 0.1, and 0.5 μg/kg varied between 87% and 98%. The method is linear from the limit of quantification 0.05 μg/kg up to 3 μg/kg levels. This method is intended for aflatoxin M1 analyses in eggs simply with minimum toxin lose, excellent recovery, and accurate results with the limit of detection 0.01 μg/kg.

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