测定花生油中黄曲霉毒素B1前处理方法的优化
目的 建立一种环保的同时适用于高效液相色谱柱后光化学衍生法(liquid chromatography post-column photochemical derivation, LC-PSD)和酶联免疫吸附筛查法(enzyme-linked immunosorbent screening, ELISA)测定花生油中的黄曲霉毒素B1的前处理净化方法。方法 花生油试样经甲醇-水溶液(70:30, V:V)提取, 经乙醚净化、冷冻离心除脂净化和免疫亲和柱净化后, 采用LC-PSD法和ELISA法测定。结果 黄曲霉毒素B1在0.1~40 ng/mL范围内均表现出良好的线性关系, LC-PSD法相关系数均大于0.999, 检出限为0.03 μg/kg, 定量限为0.1 μg/kg。ELISA法相关系数均大于0.9346, 检出限为0.1 μg/kg, 定量限为0.3 μg/kg, 均满足现行国标要求。黄曲霉毒素B1在添加水平为5 μg/kg和20 μg/kg时, LC-PSD法的加标回收率为84%~99%, 不确定度(n=6)为0.2%~3.3%, ELISA法的加标回收率为109%~124%, 不确定度(n=6)为0.3%~10.9%。结论 优化后的花生油中黄曲霉毒素B1检测前处理方法可以使检验效率提高40%以上, 有机溶剂的使用量降低50%以上, 经济效益提高50%以上, 优化方法适合大批量测定花生油中黄曲霉毒素B1。
Objective To establish an environmentally friendly pretreatment and purification method suitable for determination of aflatoxin B1 in peanut oil by liquid chromatography post-column photochemical derivation (LC-PSD) and enzyme-linked immunosorbent screening (ELISA). Methods Peanut oil samples were extracted by methanol-aqueous solution (70:30, V:V), purified by ether, freezing centrifugation and immune affinity column, then determined by LC-PSD and ELISA method. Results Aflatoxin B1 showed a good linear relationship within the range of 0.1~40 ng/mL, the correlation coefficients of LC-PSD method were all greater than 0.999, the detection limit was 0.03 μg/kg, and the quantitative limit was 0.1 μg/kg. The correlation coefficients of ELISA were all greater than 0.9346, the detection limit was 0.1 μg/kg, and the quantitative limit was 0.3 μg/kg, which met the current national standard requirements. When the addition level of aflatoxin B1 was 5 μg/kg and 20 μg/kg, the standard recovery rate of LC-PSD method was 84%?99% and the uncertainty (n=6) was 0.2%?3.3%, while the standard recovery rate of ELISA method was 109%?124% and the uncertainty (n=6) was 0.3%?10.9%. Conclusion The optimized aflatoxin B1 test pretreatment method in peanut oil can improve the test efficiency by more than 40%, reduce the use of organic solvents by more than 50%, and increase the economic benefit by more than 50%. The optimized method is suitable for the mass determination of aflatoxin B1 in peanut oil.
标题：测定花生油中黄曲霉毒素B1前处理方法的优化
英文标题：Optimization of pretreatment for determination of aflatoxin B1 in peanut oil
作者：
于金辉 河源市食品检验所
陈碧聪 河源市食品检验所
成莲 河源市食品检验所
吴丽娟 河源市食品检验所
中文关键词:黄曲霉毒素B1,前处理,酶联免疫吸附筛查法,高效液相色谱柱后光化学衍生法,
英文关键词:aflatoxin B1,pretreatment,enzyme-linked immunosorbent screening,high performance liquid chromatography post-column photochemical derivation,
发表日期：2019-08-13
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