测定花生油中黄曲霉毒素B1前处理方法的优化

Optimization of pretreatment for determination of aflatoxin B1 in peanut oil

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

中文摘要:

      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。

英文摘要:

      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.