HPLC法测定藏药二十五味鬼臼丸中鬼臼毒素的含量

目的：建立HPLC法并对二十五味鬼臼丸中的鬼臼毒素进行含量测定。方法：采用Shim-pack VPODS C₁₈（250 mm×4.6 mm，5 μm）色谱柱，以乙腈-0.2%磷酸水溶液为流动相，线性梯度洗脱（0～10 min，35%乙腈；10～25 min，35%乙腈→55%乙腈；25～30 min，55%乙腈→35%乙腈），流速1.0 mL·min^-1，检测波长290 nm，柱温30℃。结果：鬼臼毒素的线性范围为0.02～0.33 μg（r=0.999 9），方法回收率（n=6）为100.5%（RSD=0.57%）。全国7个生产企业的69批次二十五味鬼臼丸中鬼臼毒素含量范围为0.02～0.77 mg·g^-1；二十五味鬼臼丸中鬼臼毒素含量限度定为0.07 mg·g^-1。结论：本方法制定了二十五味鬼臼丸处方中主药鬼臼中鬼臼毒素的含量限度，为该药的质量控制提供了有力的依据。

Objective: To develop an HPLC method for determination of podophyllotoxin in Ershiwuwei Guijiu pills.Methods: The samples were separated on a Shim-pack VP-ODS C₁₈ column(250 mm×4.6 mm, 5 μm) with gradient elution at the flow rate 1.0 mL·min^-1.The mobile phase consisted of acetonitrile and water containing 0.2% phosphoric acid in a linear gradient elution(0-10 min,35% acetonitrile;10-25 min,35% acetonitrile→55% acetonitrile;25-30 min,55% acetonitrile→35% acetonitrile).The detecting wavelength was 290 nm,and the column temperature was 30℃.Results: Podophyllotoxin was separated excellently,the linear range of the injection amounts was from 0.02 to 0.33 μg(r=0.999 9),and the average recovery(n=6) was 100.5% with RSD of 0.57%.The content range of podophyllotoxin in 69 samples of Ershiwuwei Guijiu pills from 7 pharmaceutical companies was 0.02 to 0.77 mg·g^-1.The content limitation of podophyllotoxin in Ershiwuwei Guijiu pills was 0.07 mg·g^-1. Conclusion: The method has developed the podophyllotoxin content limit in the main prescription medicines Guijiu of Ershiwuwei Guijiu pills,which provides a strong basis for the quality control of the drug.

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