Simultaneous immunoaffinity column cleanup and HPLC analysis of aflatoxins and ochratoxin A in Spanish bee pollen

Bee pollen is a major substrate for mycotoxins growth when no prompt and adequate drying is performed by the beekeeper after collection by bees. Regulatory limits for aflatoxins and ochratoxin A are currently in force in the European Union for a rising list of foodstuffs, but not for this. An immunoaffinity column cleanup process has been applied prior to the analysis of aflatoxins B-1, B-2, G(1), and G(2) and ochratoxin A (OTA). Optimization of the HPLC conditions has envolved both a gradient elution and a wavelength program for the separation and fluorimetric quantitation of all five mycotoxins at their maximum oxcitation and emission values of wavelength in a single run. The higher limit of detection (mug/kg) was 0.49 for OTA and 0.20 for aflatoxin B-1. Repeatability (RSDr) at the lower limit tested ranged from 9.85% for OTA to 6.23% for aflatoxin G2, and recoveries also at the lower spiked level were 73% for OTA and 81% for aflatoxin B-1. None of the 20 samples assayed showed quantifiable values for the five mycotoxins.