Simultaneous determination of trace Aflatoxin B1 and Ochratoxin A by aptamer-based microchip capillary electrophoresis in food samples

Abstract

An aptamer-based microchip capillary electrophoresis coupled with laser induced fluorescence (MCE-LIF) detection method for fast determination of Aflatoxin B₁ (AFB₁) and Ochratoxin A (OTA) was developed. Aptamers that are specific to these two mycotoxins were first hybridized with their aptamer complementary oligonucleotides. The double strand DNA that comes in contact with mycotoxin-containing environment would be unwound into separate aptamer-mycotoxin complex and aptamer complementary single strand. Different types of oligonucleotides can be separated in MCE and detected under the aid of fluorescent dye SYBR gold in LIF detection unit. Under the optimal conditions, on-chip aptamer-mycotoxin conjugates analysis was achieved within 3 min with extremely low LODs (0.026 ng/mL for AFB₁ and 0.021 ng/mL for OTA). Specificity study indicated that other major mycotoxins would not cross-react with these two aptamers, demonstrating the good selectivity of the proposed method. Quantification of trace AFB₁ and OTA in real food samples was carried out and satisfactory recoveries were obtained. It is demonstrated that this method is fast, facile and specific for Simultaneous determination of trace AFB₁ and OTA from foodstuffs.

Keywords: Aflatoxin B(1); Aptamer; Laser induced fluorescence; Microchip capillary electrophoresis; Ochratoxin A.