ELISA和免疫亲和柱HPLC方法测定粮油食品中黄曲霉毒素B1

OCCURRENCE OF AFLATOXIN B1 IN RICE,FLOUR AND COOKING OIL DETERMINED BY ELISA AND IAC-HPLC.

目的:建立一套快速、灵敏、准确的测定粮油食品中AFB1的检测方法,并对市售大米、面粉、食用油样品中AFB1的污染状况进行检测分析。方法:大量样品首先用直接竞争ELISA方法初筛测定,阳性样品再用免疫亲和柱净化、荧光检测器测定的HPLC方法验证和准确定量。结果:用ELISA法和HPLC法测定食用油样品的回收率分别为74 6 %～10 9%和80 7%～98 0 % ,加标样品6次平行测定的RSDr分别为0 6 3%～2 8%和0 6 3%～2 8% ,10份阳性样品测定结果显示两种具有很好的符合性(r =0 996 8) ;16 6份样品中经HPLC法验证检出2 0份阳性,含量范围为0 8～4 8 6 μg/kg。结论:建立了一种简便、快速、高效的ELISA法初筛检测结合HPLC法验证和定量测定粮油食品中AFB1的检测方法;在抽检的样品中发现有12 %污染了AFB1,主要为花生油样品,但绝大多数阳性样品(95 % )的污染水平不高(<2 0 μg/kg)。

Objective:To investigate the occurrence of aflatoxin B1 in rice, flour and cooking oil by using direct competitive enzyme-linked immunosorbent assay (ELISA) kit and high-performance liquid chromatography (HPLC). Methods:The concentrations of AFB1 in the samples extracts were initially estimated by ELISA.Samples found to be positive with ELISA were further confirmed and quantified with HPLC.Results:The recovery rates of AFB1 from the cooking oil spiked at levels between 1 and 10 μg/kg were 74.6-109% by ELISA. By HPLC, the recovery rates of AFB1 spiked at levels between 5 and 20 μg/kg were 80.7～98.0%. Among a total of 166 samples collected in market, twenty samples resulted positive in the range of 0.8～48.6 μg/kg, the incidence of AFB1 in rice, flour and cooking oil was 9%, 5% and 18%, respectively, when determined by HPLC.Conclusion:These results obtained by HPLC were closely related to those by ELISA for AFB1(r=0.9968).Although AFB1 was confirmed in 12% of the samples, the concentrations in most these cases(95%)were found to be lower(<20 μg/kg).