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Aflatoxin M1 2000 In Milk
(Rapid Format)

Cat. No. 961AFLM01M-US-96

The chart below illustrates the difference in the two assay procedures so the user can select the appropriate format as required for the intended analysis:

................................... M1 High Sensitivity Vs. M1 2000 Rapid Format
  M1 High Sensitivity
Cat No. 961AFLM01M-96
M1 2000 Rapid Format
Cat. No. 961AFLM01M-US-96
Reaction Range 0 - 100 pg/mL(ppt) 0 - 2000 pg/mL(ppt)
Standards .... 0, 5, 10, 25, 50, 100.... .. 0, 100, 250, 500, 1000, 2000 ..
Dilution None 1 : 5
.. Limit of Detection .. 5 pg/mL(ppt) 100 pg/mL(ppt)
Assay Time 2 hr. 30 min. 35 minutes

The Aflatoxin M1 2000 is a competitive ELISA Immunoassay for the quantitative detection of Aflatoxin M1 in milk and milk powder.

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. Aflatoxin B1 is the most toxic and frequently detected form. The other types present a significant danger if the concentration is at a high level. 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 ug/L or 50 parts per trillion (50 ppt.) The equivalent value in the USA is 0.5 ug/L or 500 ppt.

Intended Use

The HELICA Aflatoxin M1 2000 Assay is intended for the quantitative detection of Aflatoxin M1 in milk, reconstituted milk powders and cheese.

Assay Principle

The HELICA Aflatoxin M1 Assay is a solid phase competitive enzyme immunoassay. An antibody with a high affinity for aflatoxin M1 is coated onto polystyrene microwells. Standard or sample is added to the appropriate well and if Aflatoxin M1 is present it will bind to the coated antibody. Subsequently, aflatoxin bound to horse -radish peroxidase (HRP) is added and binds to the antibody not already occupied by aflatoxin M1 present in the sample or standard. After this incubation period, the contents of the wells are decanted, washed and an HRP substrate is added which develops a blue color in the presence of enzyme. The intensity of the color is directly proportional to the amount of bound conjugate and inversely proportional to the amount of aflatoxin M1 in the standard or sample. Therefore, as the concentration of aflatoxin M1 in the sample or standard increases, the intensity of the blue color will decrease. An acidic stop solution is added which changes the chromogen color from blue to yellow. The microwells are measured optically by a microplate reader with an absorbance filter of 450nm (OD450). The optical densities of the samples are compared to the OD's of the kit standards and an interpretative result.


Aflatoxin M1 2000 Standards
  pg/mL Commodity ppt
1 0.0 0.0
2 100 100
3 200 200
4 500 500
5 1000 1000
6 2000 2000



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