基于超导体包被的免疫荧光快检试纸法同时测定玉米中3种毒素的适用性研究

Study on the Applicability of Simultaneous Determination of Three Mycotoxin in Maize by Superconductor Coated Immunofluorescence Rapid Detection

应用基于超导体包被的免疫荧光快速定量检测技术,建立了一种AFB1-ZEN-DON三合一联检卡法,可同时快速测定玉米中的黄曲霉毒素B1(AFB1)、玉米赤霉烯酮(ZEN)、呕吐毒素(DON)三种常见真菌毒素。对质控样准确度、加标回收率、检测限、定量限进行测定,检测多个玉米样品并与液相色谱法的检测结果进行对比,评估方法的适用性。结果表明,AFB1-ZEN-DON三合一联检卡检测AFB1含量在0.5~30μg/kg,ZEN含量在5~200μg/kg,DON含量在100~2 500μg/kg的范围内线性良好。AFB1在5~20μg/kg加标量范围时,回收率为97.62%~99.37%,ZEN在50~150μg/kg加标量范围时,回收率为98.03%~104.07%, DON在500~2000μg/kg加标量范围时,回收率为102.49%~108.53%。与液相色谱法相比,使用该方法检测AFB1相对偏差在–2.84%~11.91%,ZEN相对偏差在–3.43%~18.12%,DON相对偏差在1.83%~12.91%,均在液相色谱法允许偏差20%、15%和23%之内。该方法检测AFB1检测限为0.41μg/kg,定量限为1.12μg/kg;ZEN检测限为3.22μg/kg,定量限为9.15μg/kg;DON检测限为35.42μg/kg,定量限为100.84μg/kg。通过研究证明,AFB1-ZEN-DON三合一联检卡是一种有效、准确、快速的方法,能满足同时检测玉米中黄曲霉毒素B1、玉米赤霉烯酮、呕吐毒素的要求。

In order to rapidly determine aflatoxin B1(AFB1), zearalenone(ZEN) and deoxynivalenol(DON)in maize, a AFB1-ZEN-DON three-in-one combined detection method has been established. This quantitative immunofluorescence assay based on superconductor coating was used to detect multiple maize samples.Combined with the recovery rate, stability, detection limit, precision and other indexes, the applicability of the method were evaluated and compared with the results of liquid chromatography. The results showed that the AFB1-ZEN-DON three-in-one combined detection method had the good linearity in the range of 0.5~30 μg/mL for AFB1, 5~200 μg/kg for ZEN and 100~2 500 μg/mL for DON, respectively. The relative deviation of AFB1, ZEN and DON detected by this method was –2.84%~11.91%, –3.43%~18.12% and 1.83%~12.91%,respectively, which were within the allowable deviation of 20%, 15% and 23%. The detection limit of AFB1was 0.41 μg/kg and the corresponding limit of quantification was 1.12 μg/kg. The limit of detection of ZEN was 3.22 μg/kg and the corresponding limit of quantitation was 9.15 μg/kg. The detection limit of DON was 35.42 μg/kg and the corresponding limit of quantitation was 100.84 μg/kg. Compared with the results of liquid chromatography, the superconductor coated immunofluorescence test strip method is an effective,practical, rapid and quantitative analysis method within the allowable deviation range, suggesting that it could meet the requirements of simultaneous detection of AFB1, ZEN and DON in maize.