Determination of ochratoxin A at the ppt level in human blood, serum, milk and some foodstuffs by high-performance liquid chromatography with enhanced fluorescence detection and immunoaffinity column cleanup: methodology and Swiss data.

An improved specific analytical method for ochratoxin A (OA) is presented, combining HPLC separation with enhanced fluorescence detection by post-column addition of ammonia. Commercial immunoaffinity columns (Biocode) were for the first time applied to the cleanup of extracts of body fluids; they could be used up to 20 times for blood serum. The extraction efficiency of OA from human serum and milk as well as its derivatization to esters were studied and improved. The quantitation limit for OA was improved and estimated at 5-10 pg/g for human milk and serum. The mean recovery of OA from serum and milk was estimated at 85%. The overall coefficient of variation for OA determinations in serum, milk and selected foodstuffs was estimated at 10% (concentration range 0.01-5 ng/g). The method was applied to sera of 368 blood donors, 10 pairs of maternal and fetal sera, as well as to 40 human milk samples and selected foodstuffs; the results are discussed.