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Article Information

  • Title: Determination of Aflatoxin M1 in whey powder by HPLC and ELISA methods

  • Authors: Mahboubeh Shaneshin, Hoda Noursalehi, Zahra Mousavi, Zeinab Pourjabbara, Mannan Hajimahmoodi

  • Affiliations:

    • Drug and Food Control Department, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran

    • Food and Drug Administration, Tehran University of Medical Sciences, Tehran, Iran

    • Pharmacology & Toxicology Department, Faculty of Pharmacy & Pharmaceutical Sciences, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran

  • Journal: J Food Safe & Hyg

  • Publication Date: Winter & Spring 2018

  • Keywords: Aflatoxins; HPLC; ELISA; Whey powder

Abstract

  • Whey powder is widely used in the food industry and must be free from toxins such as Aflatoxin M1 to ensure consumer health.

  • Study aimed to determine Aflatoxin M1 levels in whey powder from two domestic factories.

  • Methodology:

    • Samples collected over 8 months (Dec 2014 - July 2015).

    • 44 samples assessed using ELISA; 26 samples analyzed via HPLC-FLD.

  • Results:

    • 44 samples (52.4%) were below the safety limit (1000 ppt).

    • 40 samples (47.6%) exceeded the maximum tolerance limit.

    • Average concentration was 1100.2 ± 734.8 ppt; range 200 - 7000 ppt.

    • No significant seasonal impact on Aflatoxin M1 levels was observed.

  • Conclusion: Contamination of whey powder with Aflatoxin M1 poses health risks.

Introduction

  • Aflatoxins:

    • Fungal toxins produced by Aspergillus species under favorable conditions (moisture, heat).

    • Found in agricultural products like peanuts and grains.

    • Types: Eighteen known types, including Aflatoxin B1 (AFB1) and its metabolite Aflatoxin M1 (AFM1).

    • AFM1 is stable and resistant to degradation during pasteurization.

  • Health Risks:

    • AFM1 is classified as a carcinogen by IARC and WHO.

    • Commonly detected in milk from cattle fed contaminated fodder.

Methods

2.1. Samples

  • 84 whey powder samples from two Iranian factories analyzed (Dec 2014 - July 2015).

  • 26 samples were further analyzed using HPLC.

2.2. Chemicals and Reagents

  • ELISA conducted using Ridascreen AFM1 kit; HPLC using HPLC-grade acetonitrile and dehydrated water.

2.3. ELISA Procedure

  • Samples prepared by dissolving 10g in 100ml water, followed by heating and centrifugation.

  • AFM1 measured using ELISA by following manufacturer's protocol.

  • Standard curve prepared to calculate concentration using various concentrations of AFM1.

2.4. HPLC Procedure

2.4.1. Sample Preparation

  • 5g whey powder mixed with water, then centrifuged to separate oil and prepare for purification.

  • Samples treated with immunoaffinity chromatography for AFM1 purification.

2.4.2. Apparatus

  • HPLC setup included Agilent 1200 series; column ZORBAX Eclipse-XDB C18.

2.5. Method Validation

  • Linearity assessed with correlation coefficients and limits of detection (LOD) and quantification (LOQ) calculated.

2.6. Statistical Analysis

  • Data analyzed using t-test and ANOVA; significance at p < 0.05.

Results

  • Calibration curves generated for AFM1 using both ELISA and HPLC methods.

  • Contamination Levels:

    • 84 samples: 44 (52.4%) less than 1000 ppt, 40 (47.6%) exceeded the limit.

    • HPLC confirmed findings; no significant difference between ELISA and HPLC results.

Discussion

  • AFM1 presents serious health risks; thorough evaluation of milk quality is essential.

  • No seasonal impact observed; this study aligns with similar research findings.

  • Identified contamination in products requires ongoing monitoring.

Conclusion

  • AFM1 detected in whey powders indicates potential health risks to consumers.

  • Recommendations include using high-quality milk to mitigate contamination.

Acknowledgements

  • Supported by a grant from the Drug and Food Control Dept., Faculty of Pharmacy, Tehran University of Medical Sciences.

References

  • Comprehensive list of studies related to AFM1 contamination, methodology, and health implications.