Development of a Time-Resolved Fluorescence Immunochromatographic Assay for the Quantitative Determination of AFB1 and ZEN in Grain
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Abstract:
A time-resolved fluorescence immunochromatographic assay for the quantitative determination of AFB1 and ZEN in grain was developed. Anti-AFB1 and anti-ZEN were labelled with time-resolved fluorescent microsphere as tracers. The assay conditions, including pH value of labelling, concentrations of antibody for labeling, concentration of the coating antigen (T line) and the goat-anti-mouse IgG (C line), as well as the sample pretreatment were optimized. Under the optimal condition, the assay showed the AFB1 limit of detection and detection range were 0.80 ng/mL and 0.81~5.67 ng/mL, respectively; the AFB1 IC50 was 2.15 ng/mL; the assay showed the ZEN limit of detection and detection range were 4.58 ng/mL and 4.76~85.60 ng/mL, respectively, the ZEN IC50 was 20.19 ng/mL. The cross-reaction rates with T-2、DON、FB1、OTA were less than 10%, indicating a good specificity of the proposed assay. Recovery test was done by using maize, soybean, wheat bran and wheat sample. The AFB1 recoveries ranged from 97.10% to 108.7% with RSDs below 15% and the ZEN recoveries ranged from 92.8% to 109.1% with RSDs below 15%. The results of the FAPAS had a good agreement with those of UPLC-MS/MS. The results had a good agreement with colloidal gold immunochromatography and ELISA. In conclusion, theproposed assay was convenient、rapid、accurate and sensitive. The detection process is about 25 min. It is suitable for rapid screening of AFB1 and ZEN in grain samples.
