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目的建立具有大体积流通池的荧光检测超高效液相色谱法测定食品中4种黄曲霉毒素(B_1、B_2、G_1、G_2)的方法。方法食品经乙腈水提取液超声提取30 min,免疫亲和柱净化,氮气吹干,10%乙腈溶液定容,C_(18)反相色谱柱分离,采用具有大体积流通池的荧光检测器检测,激发波长为365 nm,发射波长为450 nm。结果 4种黄曲霉毒素在C_(18)高效液相色谱柱上得到了很好的分离,5 min内完成检测,方法的检出限为0.010μg/kg(黄曲霉毒素B_1、黄曲霉毒素G_1)和0.003μg/kg(黄曲霉毒素B_2、黄曲霉毒素G_2),平均回收率为77.55%~102.89%,RSD为1.2%~4.4%,4种黄曲霉毒素在0.1 ng/ml~10 ng/ml时线性关系良好(相关系数r>0.999)。采用大体积流通池的荧光检测器后,灵敏度提高5倍~20倍。结论采用免疫亲和柱净化有效去除了基质中的杂质,改用大体积流通池的荧光检测器,省去了衍生步骤,简单快速,提高了黄曲霉毒素检测的灵敏度及准确度,适用于食品中黄曲霉毒素的测定。
OBJECTIVE To establish a method for the determination of four aflatoxins (B_1, B_2, G_1, G_2) in food by fluorescence detection and high performance liquid chromatography with large volumetric flow cells. Methods The food was extracted with acetonitrile by ultrasonic extraction for 30 min, purified by immunoaffinity column, dried with nitrogen, fixed in 10% acetonitrile solution and separated on a C 18 reversed-phase column using a fluorescence detector with a large volume flow cell , The excitation wavelength is 365 nm and the emission wavelength is 450 nm. Results The four aflatoxins were well separated on a C 18 high performance liquid chromatographic column and the detection limit was within 5 min. The detection limit was 0.010 μg / kg (aflatoxin B_1, aflatoxin G_1 ) And 0.003 μg / kg (aflatoxin B_2, aflatoxin G_2) with the average recoveries of 77.55% -102.89% and RSDs of 1.2% -4.4%, respectively. The four aflatoxins ranged from 0.1 ng / ml to 10 ng / ml when the linear relationship was good (correlation coefficient r> 0.999). With a large volume flow cell fluorescence detector, the sensitivity increased by 5 times to 20 times. Conclusion Immunoaffinity column purification can effectively remove the impurities in matrix and use the fluorescence detector in large-volume flow cell instead of the derivatization step. It is simple and rapid, and improves the sensitivity and accuracy of aflatoxin detection. It is suitable for food Determination of aflatoxins.