Pickering Laboratories developed an easy and sensitive method to analyze aflatoxins B1, B2, G1, G2, and ochratoxin A in hemp and hemp-containing edible products. Mycotoxins are isolated using immunoaffinity clean-up columns and analyzed with fluorescence detection. To increase sensitivity of aflatoxins B1 and G1, an in-line photochemical reactor (UVETM) is installed before the detector. This method utilizes standard HPLC equipment and allows laboratories to easily determine these mycotoxins at low ppb levels.
Isolation of aflatoxins B1, B2, G1, G2, and ochratoxin A
Blend 1 g of finely ground sample with extraction solution (10 mL of methanol/water 80:20) using a handheld homogenizer. Centrifuge for 10 min. Mix 2 mL of the extract with 10 mL of PBS buffer (containing 2% Tween 20). Clean the extracts using AflaOchra HPLC immunoaffinity column (Vicam).
Analytical Column: MycotoxTM (Pickering), C18, 4.6 × 250 mm
HPLC eluent: sodium phosphate buffer (Cat #1700–1108), methanol, acetonitrile
Flow rate: 1 mL/min
Post-column photochemical reactor: UVE (Cat # 10519 (240V), Cat # 10742 (120 V) FLD: Excitation 36 nm, Emission 430 nm for Aflatoxins Excitation 333 nm, Emission 477 nm for Ochratoxin A.
The 5-point calibration curves were built in the ranges of 0.325– 3.25 ppb for B1, 0.088–0.882 ppb for B2, 0.310–3.099 ppb for G1, 0.099–0.996 ppb for G2, and 1–10 ppb for ochratoxin A. Correlation coefficient was R2 >0.999 for all toxins.
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