Determination of ochratoxin A in cocoa beans using immunoaffinity column cleanup with high-performance liquid...

A method was developed and validated for the determination of ochratoxin A (OTA), a fungal metabolite, in cocoa beans of high fat content. The sample was extracted by blending with a 1% sodium bicarbonate solution (pH 10) followed by ultrasonication, and the sample was defatted by treatment with a flocculant. The defatted sample was purified using immunoaffinity column chromatography, and OTA was detected using HPLC with fluorescence detection. The method was fully optimized, validated, and quality controlled using spike recovery analyses, with recoveries of 89-105% over spiking ranges of 320-2.5 ng/g with CV of analyses generally <10% over 4 consecutive years and an LOQ of 0.66 ng/g in cocoa bean samples. This method overcomes the problems posed by the high fat contents of cocoa and chocolate samples with a high degree of reliability.