C/N: EE 102

1. BACKGROUND

Aflatoxins are carcinogenic, highly toxic metabolites of the mold fungus varieties Aspergillus flavus and Aspergillus parasiticus. Aflatoxin M1 is produced as a metabolite of aflatoxin B1. It is secreted with the milk after feeding of aflatoxin B1 containing feed to lactating cows. As aflatoxin M1 is relatively stable towards the pasteurizing process, not only a comprehensive routine check of the raw materials to be processed is required, but also of the final products.

2. PURPOSE

This handbook establishes procedures for determining aflatoxin M1 in milk ,milk powder, yogurt and cheese products. The test is a competitive ELISA that provides exact concentrations in parts per billion (ppb).The basis of the test is the antigen-antibody reaction. The wells in the microtiter are coated with specific antibodies against aflatoxin M1.Aflatoxin M1 standards or the sample solutions and after a washing step the enzyme conjugate are added. Free aflatoxin M1 and aflatoxin M1 enzyme conjugate compete for the aflatoxin M1 antibody binding sites. Any unbound enzyme conjugate is then removed in a washing step. After a wash step, substrate reacts with the bound enzyme conjugate to produce blue color. A stop solution is then add which changes the color from blue to yellow. The intensity of the color is inversely proportional to the concentration of aflatoxin M1 in the samples and standards.

3. KIT CONTENTS

1) Microtiter plate (8wells×12): precoated with antibodies.

2) Aflatoxin M1 standard solutions (1mL/each):0ng/mL, 0.05ng/mL, 0.1ng/mL,0.2ng/mL, 0.5ng/mL

3) AF-M1 Enzyme Conjugate: (15mL)

4) Washing buffer (10×): (50mL)

5) AF-M1 Diluent Buffer: (50mL)

6) TMB: (12mL)

7) Stop solution：(7mL)

8) Product Manual

9) COA report