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Aflatoxin M1 in Milk

Aflatoxin M1 in Milk, Milk Powder and Cheese

Published: August 8, 2008
By: Christine Thornton - Helica Biosystems, Inc.
Aflatoxins are toxic metabolites produced by a variety of molds such as Aspergillus flavus  and Aspergillus parasiticus. They are carcinogenic and can be present in grains, nuts, cottonseed and other commodities associated with human food or animal feeds. Crops may be contaminated by one or more of the four following sub-types of aflatoxin: B1, B2, G1 and G2. While the other sub-types of aflatoxin present a significant danger at high concentration levels, aflatoxin B1 is the most toxic and frequently detected form.


Aflatoxins have been implicated in human health disorders including hepatocellular carcinoma, aflatoxicosis, Reye’s syndrome and chronic hepatitis. Animals are exposed to aflatoxins by consumption of feeds that are contaminated by aflatoxin-producing fungal strains during growth, harvest or storage. When cows are fed contaminated feed, aflatoxin B1 is converted by hydroxylation to aflatoxin M1, which is subsequently secreted in the milk of lactating cows. Aflatoxin M1 is quite stable towards the normal milk processing methods such as pasteurization and if present in raw milk, it may persist into final products for human consumption.


Most controlling government agencies worldwide have regulations regarding the amount of aflatoxins allowable in human and animal foodstuffs. Many countries have declared limits for the presence of aflatoxin M1 in milk and milk products. In the EU the limit for the presence of M1 in milk and reconstituted milk powders has been set at 0.05 mg/L or 50 parts per trillion (50 ppt.)


In response to the EU’s growing demand for stringent regulation of aflatoxin M1 in milk products, Helica Biosystems has created quantitative ELISA assays for the detection and quantization of aflatoxin M1 in milk, reconstituted milk powders and cheese. Detection of aflatoxin M1 is a simple process, with just a single dilution step and no complicated extraction.


Recovery and Reproducibility


Recovery of 50 ppt spiked into milk is as follows:


 

% Recovery

Within Assay CV

SKIM MILK
                    50 ppt


100


4.2%

1% FAT HOMOGENIZED
                    50 ppt


93


4.4%

FULL FAT HOMOGENIZED
                    50 ppt


92


2.2%




Recovery in cheese: 100 pg of aflatoxin M1 was spiked into 1 gm of finely grated parmesan cheese and allowed to remain in contact for one hour at ambient temperature. Applying the extraction procedure described above, the recovery of aflatoxin M1 was 60.5% with a CV of 5.5% for 8 separate spiking/extractions.


Performance Data


Sensitivity

N

Mean OD

SD

% CV

Sensitivity

18

1.857

0.023

1.2

2 ppt



Precision

Intra-assay

Sample
(ppt)

N

Mean OD

% B/B 0

% CV

0

8

1.245

100

1.4

5

8

1.092

87.7

2.0

10

8

0.945

75.9

3.0

25

8

0.719

57.8

2.1

50

8

0.449

36.1

2.7

100

8

0.246

19.8

2.0

Over a seven-month period with multiple lots

Inter-assay

Sample
(ppt)

N

% B/B 0

   % CV  

5

8

86.6

1.5

10

8

76.2

3.3

25

8

55.5

4.7

50

8

36.3

4.8

100

8

22.0

10.2



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Hernan Farias Chacon.
7 de octubre de 2013
Appreciate anyone that could contribute with a standard limit of Aflatoxin in Milk Powder valid for international trade. The EU adopt 0.05 u/kg, apparently different US Institution adopted 0.5 u/kg, hence further clarification is highly appreciated. Hernan Farias QA Manager in KSA
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Dr.g.sarathchandra
Dr.g.sarathchandra
28 de agosto de 2008
What is the detection limit? Why cant it be attained by lc?
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Cor Arts
Arts Project Support
11 de agosto de 2008
Sensitivity is given for only one product. It is not given which product. The recovery of spiked Aflatoxine M1 from cheese is too low. It is advised to look for another and more effective extraction method. Best regards, Cor
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