青蒿素有关物质检查方法探讨

目的：建立青蒿素有关物质含量的HPLC检查方法。方法：采用高效液相色谱法对青蒿素原料中有关物质进行定量测定，以CAPCELL PAK C₁₈ MG（4.6 mm×150 mm，5 μm）为色谱柱，以乙腈-水（60：40）为流动相，流速为1.0 mL·min^-1，检测波长为210 nm；采用超高效液相色谱-三重四极杆质谱联用技术（UPLC-MS/MS）对实测样品中的主要杂质进行结构确认，离子源为ESI，检测模式为正离子模式。结果：主峰与各杂质分离良好，样品中检测出2个主要杂质。其中，杂质Ⅰ为青蒿烯，杂质Ⅱ可能为青蒿素的异构体。所建立的HPLC方法，青蒿素和青蒿烯的检出限分别为2 μg·mL^-1和0.05 μg·mL^-1，线性范围分别为0.01~0.12 mg·mL^-1和0.000 5~0.01 mg·mL^-1；青蒿烯的平均加样回收率为101.4%，RSD为1.8%；青蒿烯和杂质Ⅱ进样精密度分别为0.6%和1.7%。结论：本法经方法学验证，可用于青蒿素中有关物质的测定。

Objective: To establish an HPLC method for determination of the related substances in artemisinin.Methods: HPLC was employed to quantify the main impurities.A CAPCELL PAK C₁₈ MG(4.6 mm×150 mm, 5 μm)column was adopted,the mobile phase was acetonitrile-water(60∶40),the flow rate was 1.0 mL·min^-1 ,and the detection wavelength was 210 nm.LC-MS/MS was employed to identify the main impurities with an electro spray ionization(ESI)source in the positive ion mode.Results: Main peak and impurities were well separated under the established HPLC condition.Two impurities were detected and characterized by LC-MS/MS.The impurityⅠis artemisitene and the impurityⅡis probably one isomeric with artemisinin.Under optimized HPLC conditions,the detection limits(LOD,S/N=3)for artemisinin and artemisitenewere 2 μg·mL^-1 and 0.05 μg·mL^-1,and linear ranges were 0.01-0.12 mg·mL^-1and 0.000 5-0.01 mg·mL^-1,respectively.The recoveries of spike samples of artemisitene was 101.4%,the RSD was 1.8%.The injection respectabilities for artemisitene and impurityⅡwere 0.6% and 1.7%,respectively.Conclusion: It is proved by methodology validation that the established method can determine the related substances in artemisinin.

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