Identification and Estimation of Aflatoxin M1 in Milk Powder, Pasteurized Milk, and Raw Milk from the Local Market Lahore by ELISA: Aflatoxin M1 in Milk Powder, Pasteurized Milk, and Raw Milk by ELISA

Muhammad Khalid Saeed; Naseem Zahra; Hafiza Kainat Anwar; Amara Khan; Muhammad Zeeshan Kazim; Ayesha Ijaz; Ijaz Ahmad

doi:10.54393/mjz.v7i1.211

Authors

Muhammad Khalid Saeed Food and Biotechnology Research Center, Pakistan Council of Scientific and Industrial Research Laboratories Complex, Lahore, Pakistan
Naseem Zahra Food and Biotechnology Research Center, Pakistan Council of Scientific and Industrial Research Laboratories Complex, Lahore, Pakistan
Hafiza Kainat Anwar Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore, Pakistan
Amara Khan Institute of Food Science and Nutrition, Gomal University, Dera Ismail Khan, Pakistan
Muhammad Zeeshan Kazim Institute of Food Science and Nutrition, Gomal University, Dera Ismail Khan, Pakistan
Ayesha Ijaz Department of Nutrition and Health Promotion, University of Home Economics, Lahore, Pakistan
Ijaz Ahmad Food and Biotechnology Research Center, Pakistan Council of Scientific and Industrial Research Laboratories Complex, Lahore, Pakistan

DOI:

https://doi.org/10.54393/mjz.v7i1.211

Keywords:

Raw Milk, Branded Milk, Aflatoxin M1, ELISA

Abstract

Because milk has a great nutritional content and can be utilized to make functional dairy products, it is regarded as the ideal natural food for consumers of all ages. It does, however, carry the greatest danger of contaminating human diets with aflatoxin M1. The primary oxidized metabolite of the carcinogenic aflatoxin B1 is aflatoxin M1. It may be caused by feed carry-over contamination and is present in milk and other dairy products. Objectives: To measure the amount of aflatoxin M1 in samples of raw and branded milk that were obtained from the Lahore local market. Methods: Due to its excellent sensitivity, selectivity, and user-friendliness, the ELISA approach was used to measure aflatoxin M1 in milk for regulatory compliance. Results: The results of the investigation showed that AFM1 was found in seven out of ten samples of raw milk, two out of five samples of pasteurized liquid milk, and three out of five samples of brands of milk powder. Aflatoxin levels in positive samples ranged from 0.06 to 0.35 parts per billion. Additionally, validated, the ELISA kits demonstrated high recovery rates and comparable performance. Additionally, it was discovered that the Limit of Detection (LOD=0.01 ppb) levels were substantially lower than the Maximum Residue Limit (0.5ppb). Conclusion: From these results, it was concluded that the majority of milk samples were found to be contaminated with aflatoxin M1.

References

Rehman A, Jingdong L, Chandio AA, Hussain I. Livestock Production and Population Census in Pakistan: Determining Their Relationship with Agricultural GDP Using Econometric Analysis. Information Processing in Agriculture. 2017 Jun; 4(2): 168-77. doi: 10.1016/j.inpa.2017.03.002.

Musa M and Mustafa MI. Livestock Feeds and Feeding Practices in Pakistan. Livestock Feeds and Feeding Practices in South Asia. 2020; 129(2): 3.

Nasir L ans Gemede HF. Aflatoxin M1 Concentrations, Adulterants, Microbial Loads, and Physicochemical Properties of Raw Milk Collected from Nekemte City, Ethiopia. The Scientific World Journal. 2024; 2024(1): 3796985. doi: 10.1155/2024/3796985.

Saeed MK, Zahra N, Anwar HK, Anjum S, Khan A, Saeed A et al. Nutritional, Antioxidative and Aflatoxin Safety Assessment of Ginger-Turmeric Tea: A Functional Herbal Beverage Tea: Antioxidative and Aflatoxin Safety Assessment of Ginger-Turmeric Tea. Diet Factor (Journal of Nutritional and Food Sciences). 2025 Sep: 06-11. doi: 10.54393/df.v6i3.176.

Tanveer H, Glesener H, Su B, Bolsinger B, Krajmalnik-Brown R, Voth-Gaeddert LE. Evaluating Methods for Aflatoxin B1 Monitoring in Selected Food Crops Within Decentralized Agricultural Systems. Toxins. 2025 Jan 17(1): 37. doi: 10.3390/toxins17010037.

Kassa A, Talema A, Ketsela G. Determination of Aflatoxin M1 in Raw Cow's Milk by Using HPLC-FLD, in Injibara Town, Awi Zone, Amhara, Ethiopia. 2020. doi: 10.20944/preprints202010.0181.v1.

Zahra N, Saeed MK, Ahmed Z, Zain MU, Jalal T. Aflatoxin M1 in Raw Milk in Summer Season in Lahore, Pakistan. Bangladesh Journal of Scientific and Industrial Research. 2024 Jun; 59(2): 115-22. doi: 10.3329/bjsir.v59i2.69303.

Vitalini S, Iriti M, Vallone L. Mycotoxins in European Union Regulations (2023-2025). Italian Journal of Food Safety. 2025 Nov; 15(1): 14096. doi: 10.4081/ijfs.2025.14096.

Ahmad U, Ajaz H, Perveez A, Azam SE. Assessment of Aflatoxin M1 (AFM1) in Milk from Chakwal City Using ELISA Technique. Agrobiological Records. 2025 Mar; 19(11): 92-99.

Ünlü E, Saltoğlu GT, Ünlü G, Yildiz HT. Determining the Aflatoxin M1 Levels in Breast Milk. European Review for Medical and Pharmacological Sciences. 2023 Apr; 27(8).

Zahra N, Jamil N, Ahmad SR, Kalim I, Saeed MK, Ijaz A, Munir S. Assessment of Milk Quality by Analyzing Aflatoxin M1 in Branded and Non-Branded Milk Samples Collected from Different Shops/Areas in Lahore, Pakistan. Pakistan Journal of Zoology. 2020 Feb; 52(1): 373. doi: 10.17582/journal.pjz/2020.52.1.sc2.

Kafle S, Paudel M, Shrestha C, Kathayat KB, Sapkota RC, Tiwari A et al. Aflatoxin M1 Contamination in Dairy Milk in Kathmandu, Nepal. Toxins. 2024 Nov; 16(11): 468. doi: 10.3390/toxins16110468.

Zentai A, Jóźwiak Á, Süth M, Farkas Z. Carry-Over of Aflatoxin B1 from Feed to Cow Milk—A Review. Toxins. 2023 Mar; 15(3): 195. doi: 10.3390/toxins15030195.

Kourti D, Angelopoulou M, Petrou P, Kakabakos S. Sensitive Aflatoxin M1 Detection in Milk by ELISA: Investigation of Different Assay Configurations. Toxins. 2024 Nov; 16(12): 515. doi: 10.3390/toxins16120515.

Maggira M, Ioannidou M, Sakaridis I, Samouris G. Determination of Aflatoxin M1 in Raw Milk Using an HPLC-FL Method in Comparison with Commercial ELISA Kits—Application in Raw Milk Samples from Various Regions of Greece. Veterinary Sciences. 2021 Mar; 8(3): 46. doi: 10.3390/vetsci8030046.

Tarannum N, Nipa MN, Das S, Parveen S. Aflatoxin M1 detection by ELISA in Raw and Processed Milk in Bangladesh. Toxicology Reports. 2020 Jan; 7: 1339-43. doi: 10.1016/j.toxrep.2020.09.012.

Hajmohammadi M, Valizadeh R, Naserian A, Nourozi ME, Rocha RS, Oliveira CA. Composition and Occurrence of Aflatoxin M1 in Cow's Milk Samples from Razavi Khorasan Province, Iran. International Journal of Dairy Technology. 2020 Feb; 73(1): 40-5. doi: 10.1111/1471-0307.12661.

Gonçalves BL, Gonçalves JL, Rosim RE, Cappato LP, Cruz AG, Oliveira CA et al. Effects of Different Sources of Saccharomyces Cerevisiae Biomass on Milk Production, Composition, and Aflatoxin M1 Excretion in Milk from Dairy Cows Fed Aflatoxin B1. Journal of Dairy Science. 2017 Jul; 100(7): 5701-8. doi: 10.3168/jds.2016-12215.

Costamagna D, Gaggiotti M, Chiericatti CA, Costabel L, Audero GM, Taverna M et al. Quantification of Aflatoxin M1 Carry-Over Rate from Feed to Soft Cheese. Toxicology Reports. 2019 Jan; 6: 782-7. doi: 10.1016/j.toxrep.2019.07.004.

Ismail A, Akhtar S, Levin RE, Ismail T, Riaz M, Amir M. Aflatoxin M1: Prevalence and Decontamination Strategies in Milk and Milk Products. Critical Reviews in Microbiology. 2016 May; 42(3): 418-27. doi: 10.3109/1040841X.2014.958051.