Fluorescence polarization immunoassay based on a monoclonal antibody for the detection of ochratoxin A

A fluorescence polarization immunoassay (FPIA) based on a monoclonal antibody for the determination of ochratoxin A (OTA) was developed. Fluorescein-labelled OTA derivative (tracer) was synthesized and purified by thin-layer chromatography. The optimized OTA FPIA had a dynamic range from 5 to 200 ng mL(-1) with IC50 value of 30 ng mL(-1) and a detection limit of 3 ng mL(-1). The method developed was characterized by high specificity and reproducibility. Cross-reactivity with other mycotoxins (zearalenone, aflatoxins, patulin and T-2 toxin) was negligible (<0.1%). Methanol extracts of barley samples were used for the analysis. The results of OTA determination in barley were compared with those determined by indirect competitive enzyme-linked immunosorbent assay (ELISA). Recoveries for the samples spiked at 50, 100 and 500 ng g(-1) levels were 91, 90 and 97%, respectively, for FPIA, and 98, 98 and 102%, for ELISA. Naturally contaminated barley samples were analysed by these methods but some disagreement was observed between the results. The FPIA method can be applied for screening of food samples for OTA residues without a complicated clean-up.