HPLC柱后光衍生荧光法测定中药饮片中黄曲霉毒素残留量

目的: 采用免疫亲和净化HPLC柱后紫外光化学衍生荧光检测法研究中药饮片中黄曲霉毒素测定的加样回收率,考察该方法在中药饮片黄曲霉毒素残留测定中的可行性,评价120批中药饮片的污染状况,总结污染规律。 方法: 样品经溶剂提取、免疫亲和柱净化后由HPLC柱后光衍生荧光检测法测定其中黄曲霉毒素残留量,对每种饮片进行4 μg·kg^-1(以黄曲霉毒素B₁计)的加样回收率考察。 结果: 黄曲霉毒素B₁、B₂、G₁、G₂的最低检测限分别为0.44,0.12,0.41,0.16 pg;线性范围分别为1.74~218.0,0.50~62.17,1.63~203.1,0.62~61.94 pg(r﹥0.9999)。所考察饮片中黄曲霉毒素B₁、 B₂、 G₁、 G₂总回收率和B₁回收率均在60%~120%之间的占87%;被污染的批次占9.2%,其中污染程度超过目前中国药典限度的占6.7%。 结论: 免疫亲和净化HPLC柱后光衍生荧光检测法具有方法简便,衍生反应稳定,灵敏度高,重现性好,反应系统污染少的优点,适合于大多数中药饮片的黄曲霉毒素检测;种子类被污染的程度严重,应扩大该类中药的监测范围。

Objective: To investigate the recoveries of aflatoxin(AF) residues analysis of traditional Chinese medicine(TCM) Yinpian using immunoaffinity column HPLC method with post column photochemical derivatization and fluorescence detection,and evaluate the feasibility of the method.To evaluate the contamination status of 120 batches of TCM Yinpian and summarize the rule of contamination. Methods: Samples were prepared through organic solvent extraction and cleaned up with immunoaffinity column sequentially;the recoveries of aflatoxins were then determined by adding aflatoxins standard mixture at the level of 4 μg·kg^-1 according to aflatoxin B₁ content.Finally,aflatoxins B₁,B₂,G₁ and G₂ residue amount in TCM Yinpian was determined. Results: The limit of detection of aflatoxins B₁,B₂,G₁,G₂ was 0.44,0.12,0.41,0.16 pg,respectively.Aflatoxins B₁,B₂,G₁,G₂ showed good linear relationships at the ranges of 1.74-218.0 pg,0.50-62.17 pg,1.63-203.1 pg and 0.62-61.94 pg,respectively,r﹥0.9999.The recoveries of total aflatoxins and aflatoxin B₁ in 87% detected samples were between 60%-120%.9.2% of TCM Yinpian was detected to be contaminated by aflatoxins,and aflatoxins amount in 6.7% exceeded the limit of Chinese Pharmacopoeia of the People's Republic of China 2010. Conclusion: The method showed convenient operation,stable derivation reaction,high sensitivity,good repeatability and little pollution,which meets the requirement of aflatoxin residues determination in most TCM Yinpian.The seeds are easy to be contaminated,so the aflatoxins control in those TCM should be expanded.