HPLC-柱后光化学衍生法测定72批舒筋活血丸中黄曲霉毒素G₂、G₁、B₂、B₁的含量

目的：建立HPLC－柱后光化学衍生法测定舒筋活血丸中黄曲霉毒素G₂、G₁、B₂、B₁的含量，并通过对72批样品检测结果的分析初步考察舒筋活血丸制剂在黄曲霉毒素污染方面的安全性，以提示企业选择合格安全的原料药材投料。方法：采用Diamonsil C₁₈（4.6 mm×250 mm，5 μm）色谱柱，以乙腈-甲醇-水为梯度洗脱流动相，流速1.1 mL·min^-1，柱温40 ℃；采用柱后光化学衍生法，荧光检测器激发波长λ_ex=360 nm、发射波长λ_em=450 nm。结果：黄曲霉毒素G₂、G₁、B₂、B₁分别在2.806~16.825 pg（r=0.9973）、9.830 ~58.983 pg（r=0.9998）、2.891~17.345 pg（r=0.9956）、9.9~59.4 pg（r=0.9966）范围内线性关系良好；黄曲霉毒素B₁及总量回收率（n=6）分别为78.5%（RSD=1.6%）和85.5%（RSD=4.0%）。有27批样品检出黄曲霉毒素，其总量含量为0.007×10^-3~39.87×10^-3μg·g^-1。结论：该方法经方法学验证可用于舒筋活血丸中黄曲霉毒素检测；建议增订舒筋活血丸中黄曲霉毒素检查项；建议企业重视动物类药材的质量和安全性。

Objective: To establish a method for the determination of aflatoxin(AF)G₂,G₁,B₂ and B₁ by HPLC with post-column photochemical derivation in Shujin Huoxue pills.Through the analysis of test results for 72 batches of samples,the safety of Shujin Huoxue pills was preliminarily investigated in terms of aflatoxin pollution,to help manufacturers in the selection of qualified and safe raw material medicines for production. Methods: HPLC with post-column photochemical derivation method was used to detect the aflatoxin content in Shujin Huoxue pills.Diamonsil C₁₈(4.6 mm×250 mm,5 μm)chromatographic column was adopted.A gradient elution program of acetonitrile-methanol-water was adopted at the flow rate of 1.1 mL·min^-1.The column temperature was maintained at 40 ℃.The fluorescence detector λ_ex was set at 360 nm and λ_em was 450 nm. Results: The linear ranges of aflatoxin G₂,G₁,B₂ and B₁ were 2.806-16.825 pg(r=0.9973),9.830-58.983 pg(r=0.9998),2.891-17.345 pg(r=0.9956) and 9.9-59.4 pg (r=0.9966),respectively.The recovery of aflatoxin B₁ and the total recovery(n=6) were 78.5% (RSD=1.6%) and 85.5%(RSD=4.0%).27 batches of samples contained aflatoxin,and the total content was 0.007×10^-3-39.87×10^-3μg·g^-1. Conclusion: This method is suitable for the determination of aflatoxin in Shujin Huoxue pills by methodology validation.It is advisable to add the determination of aflatoxin in the quality control of Shujin Huoxue pills.And manufacturers should attach importance to the quality and safety of the traditional Chinese medicine of animal sources.