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

关键词:  高效液相色谱法  20(S)-原人参二醇  原料药  含量测定  有关物质

Key words:  HPLC  20(S)-protopanaxadiol  bulk drug  content determination  related substances

摘要:

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5 μL。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。

Abstract:

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 5 μL. 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.