Aflatoxin M1 ELISA Kit

Full Name: Aflatoxin M1 ELISA Kit
Sample Type: Milk, Milk products
Sensitivity: < 10 pg/ml

BACKGROUND

Aflatoxin M1 is a metabolite of aflatoxin B1 and can be found in milk and milk related products, minor mycotoxin. It is a member of the difurocoumarocyclopentenone series family, polycyclic aromatic compounds consisting of a cyclopenten-2-one ring that is fused to the difurocoumarin skeleton via a coumarin moiety. The use in animal feeds has been banned in Europe since 2006. In the United States, aflatoxin M1 is listed as a “restricted use” food additive. It is not approved for any human food products. It is found mostly in milk for example by-products from dairy cows, including cheese and powdered milk that contains residual ingredients such as whole or skim milk, whey protein isolates, casein, lactose, lactalbumin and salt. Chronic exposure leads to hepatic adenomas (benign tumors that gradually enlarge with time) in mice exposed to low doses during their lifetime.

The following assay is reliable and accurate protocol which is frequently being used for many applications. Aflatoxin M1 have been widely linked to liver damage and cancer which is the result of activating P450 to the epoxide causing the DNA to alkylate. The eventual toxicity is dependent on the balance of the liver enzymes that either activate it (Cytochrome P450) or those that detoxify it (Glutathione-S-Transferase).

INTENDED USE

Aflatoxin M1 ELISA kit is a quantitative procedure for analysing aflatoxin M1 levels present in milk or milk products. This assay has a minimum analytical sensitivity limit of < 10.0 pg/ml.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

Microtiter Plate: Coated with aflatoxin conjugate.
Aflatoxin M1 Standards 1-6 (10x): Concentration 0, 100, 500, 1000, 5000, 10000 pg/ml.
Anti-Aflatoxin M1 Antibody (Rabbit).
Conjugate (anti-rabbit-IgG-HRP).
Substrate Solution (TMB).
Stop Solution (0.5 M H2SO4).
Sample/Standard Diluent (PBS).
Washing Solution (10x Concentrate).
Instruction Manual.

SENSITIVITY

The minimum detection sensitivity level of aflatoxin M1 using current aflatoxin M1 ELISA kit was less than 10.0 pg/ml. The standard range for this assay is 10.0 – 1000.0 pg/ml.

ASSAY CHARACTERISTICS

– Sensitivity: < 10 pg/ml
– Recovery: Milk (102%), Cheese (60%)
– Intra-Assay Variation: 3.0% was determined
– Cross Reactivity: Relative to Aflatoxin M1 (=100%), Aflatoxin B1 (10%), Aflatoxin G1 (5%), Aflatoxin B2 (3.5%), Aflatoxin G2 (2.1%), Sterigmatocystin (0.3%).

REFERENCES

A competitive immunoassay with a surrogate calibrator curve for aflatoxin M1 in milk. Anal Chim Acta. (2011) 703 (1): 64-9. Guan D., et al.
Development of ultrasensitive direct chemiluminescent enzyme immunoassay for determination of aflatoxin-M1 in milk. Food Chem. (2014) 158: 310-4. Vdovenko M.M., et al.
Detection of aflatoxin M1 in milk by dynamic light scattering coupled with superparamagnetic beads and gold nanoprobes. J Agric Food Chem. (2013) 61 (19): 4520-5. Zhang Z., et al.
Comparison of ELISA, HPLC-FLD and HPLC-MS/MS Methods for Determination of Aflatoxin_M1 in Natural Contaminated Milk Samples. Acta Chim Slov. (2016) 63 (4): 747-756. Kos J., et al.
Direct enzyme-linked immunosorbent assayfor determining aflatoxin-M1 at picogram levels in dairy products. J Assoc Off Anal Chem. (1984) 67 (6): 1098-101. Fremy J.M. and Chu F.S.

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