基于HPLC-ELSD指纹图谱与化学计量学相结合的舒眠胶囊质量评价

Quality Evaluation of Shumian Capsules Based on HPLC-ELSD Fingerprint and Stoichiometry

关键词:  舒眠胶囊  HPLC-ELSD  指纹图谱  化学计量学  质量评价

Key words:  Shumian capsules  HPLC-ELSD  fingerprint  stoichiometry  quality evaluation

摘要:

目的 建立舒眠胶囊HPLC-ELSD指纹图谱与化学计量学相结合，全面、系统评价舒眠胶囊质量的方法。方法 采用ACE Excel AR C₁₈色谱柱（4.6 mm×250 mm，5.0 μm），流动相为乙腈-水，梯度洗脱，柱温30℃，流速为1.0 mL·min^-1，ELSD漂移管温度50℃，N₂流速为3.0 mL·min^-1。用HPLC-ELSD对15批次舒眠胶囊进行分析，建立指纹图谱共有模式，采用聚类分析（cluster analysis，CA）、主成分分析（principal component analysis，PCA）等化学计量学方法进行区分与比较，寻找导致不同批次间差异的主要标志性成分。结果 共确定了16个共有峰，归属了10个共有峰的药材来源，通过对照品比对指认了4个共有峰归属，分别为柴胡皂苷a、芍药内酯苷、芍药苷、槲皮苷，样品相似度均>0.90，CA将样品分为3类，PCA筛选出不同批次间差异的主要标记物，分别为3（芍药苷），14，15号峰。结论 本研究建立的HPLC-ELSD指纹图谱与HPLC-DAD指纹图谱相结合能对舒眠胶囊产品一致性、稳定性进行全面有效评价，可用于舒眠胶囊的质量综合评价。

Abstract:

OBJECTIVE To establish the HPLC-ELSD fingerprint of Shumian capsules and combine it with stoichiometry to comprehensively and systematically evaluate the quality of Shumian capsules. METHODS The chromatographic column was ACE Excel AR C₁₈(4.6 mm×250 mm, 5.0 μm), the mobile phase was acetonitrile-water for gradient elution at a flow rate of 1.0 mL·min^-1, the columm temperature was 30℃, ELSD evaporator tube temperature was 50℃ and the flow of N₂ was 3.0 mL·min^-1. Fifteen batches of samples were determined by HPLC-ELSD and a common mode of fingerprint was established. Based on the results, stoichiometry methods such as cluster analysis(CA), principal component analysis(PCA) were used to find main marker that caused the difference of the preparation. RESULTS There were 16 common peaks, then 10 peaks were tracked assigned and four peaks were identified as saikosaponin a, albiflorin, paeoniflorin and quercitrin. Similarities of samples were > 0.90. The CA divided the samples into three categories, and three main markers for differences between different batches, peake 3(paeoniflorin), 14 and 15 were screen out by PCA. CONCLUSION The established HPLC-ELSD fingerprint combine with HPLC-DAD fingerprint in this study can comprehensively and effectively evaluate the consistency and stability of Shumian capsules, and can provide a quality evaluation reference of Shumian capsules in the future.