HPLC测定20(S)-原人参二醇原料药中有关物质

Determination of Related Substances in 20(S)-Protopanaxadiol Bulk Drug by HPLC

目的建立一种用于分离和测定20(S)-原人参二醇原料药中的10种潜在相关杂质的高效液相色谱法。方法采用ACQUITY UPLC BEH C18色谱柱,以超纯水(A)-乙腈(B)为流动相,梯度洗脱,流速为0.20~0.25 mL·min^(–1),检测波长为203 nm,柱温为35℃,进样量5μL。结果20(S)-原人参二醇与各杂质均达到有效分离,在一定范围内线性关系良好;加样回收率(n=9)在94.8%~102.2%,RSD值均<2.0%;日内和日间精密度RSD值均<2.0%,精密度良好;杂质C、杂质F、杂质H、杂质I在室温下放置48 h内RSD值<2.0%,溶液稳定;杂质A、杂质B、杂质D、杂质E、杂质G、杂质J在室温下放置48 h内RSD值>2.0%,溶液不稳定。结论此次建立的方法简便、灵敏、准确,可同时测定原料药20(S)-原人参二醇中10种有关物质,可有效地鉴定原料药中有关物质,为其生产工艺的提高和质量控制提供参考。

OBJECTIVE To establish a high-performance liquid chromatographic method for the detection of 10 related substances in 20(S)-protopanaxadiol bulk drug.METHODS Used an ACQUITY UPLC BEH C18 chromatographic column with ultrapure water(A)-acetonitrile(B)as mobile phase.The flow rate was 0.20−0.25 mL·min^(–1),the detection wavelength was set at 203 nm,the column temperature was maintained at 35℃,and the injection volume was 5μL.RESULTS 20(S)-protopanaxadiol and various impurities had reached an effective separation with a good linear relationship;the sample recovery(n=9)was ranging from 94.8%to 102.2%.The RSD values were<2.0%;the intra-day and inter-day precision RSD values were<2.0%,with a good precision;the related substances C,F,H,and I were stable in solution with RSD values<2.0%in 48 h at the room temperature;while impurities A,B,D,E,G and J were unstable under the same condition with RSD>2.0%.CONCLUSION The method is simple,sensitive and accurate,which can simultaneously determine 10 related substances in API 20(S)-protopanaxadiol,which can effectively identify the relevant substances in API and provide reference for the improvement of its production process and quality control.