The invention discloses an HPLC-FLD pre-column
derivatization method for simultaneously determining aflatoxins B1, B2, G1 and G2 in tobacco. The HPLC-FLD pre-column
derivatization method comprises the following steps: performing water-bath ultrasonic extraction on a tobacco
powder sample by use of a
methanol solution, filtering, diluting the filtrate and enabling the diluted filtrate to pass through an immunoaffinity column, performing twice drip washing on the immunoaffinity column by use of pure water, next, eluting four aflatoxins enriched on the immunoaffinity column by use of
methanol, performing blow-
drying on the eluted solution by use of
nitrogen, then, adding
trifluoroacetic acid and normal
hexane for
derivatization at the constant temperature being 45 DEG C for 40 minutes, adding a small quantity of
methanol solution and carrying out HPLC-FLD analysis on the taken low-layer solution. The HPLC-FLD pre-column derivatization method can be applied to simultaneous determination of the aflatoxins B1, B2, G1 and G2 in tobacco and tobacco products, and is capable of completing determination in 20 minutes; the four targets can be well separated, the
linear relation is good and the
detection limit is low; besides, the HPLC-FLD pre-column derivatization method has good
recovery rate and reproducibility, and is suitable for detecting the aflatoxins in tobacco and tobacco products.