基于HPLC-CAD指纹图谱结合化学计量分析评价牛黄清心丸(局方)质量

Quality evaluation of Niuhuang Qingxin pill(prescription of the bureau)by HPLC⁃CAD fingerprint combined with stoichio⁃metric analysis

目的 建立牛黄清心丸(局方)的指纹图谱,并通过化学计量学分析手段对影响其质量的关键指标成分进行分析,为牛黄清心丸(局方)的质量控制与评价提供参考。方法 采用高效液相色谱法-电雾式检测器建立牛黄清心丸(局方)指纹图谱,进行方法学考察,并确定其共有峰进行聚类分析、主成分分析和正交偏最小二乘法判别分析。结果 指纹图谱方法学考察结果显示,精密度、稳定性、重复性的相对保留时间和相对峰面积的RSD分别为0.04%~0.06%和1.45%~2.67%,0.04%~0.87%和0.26%~2.87%,0.01%~0.34%和1.72%~2.93%。对照指纹图谱共标记出26个共有峰,其中指认出苦杏仁苷、甘草苷、黄芩苷、胆酸等18个特征峰,相似度为0.927~0.995。聚类分析热图将样品聚为3类,经主成分分析得6个主要成分的累计贡献率到达84.952%,并通过正交偏最小二乘法判别分析预测得到甘草苷、甘草酸铵、升麻素苷、苦杏仁苷4个引起质量差异性的化合物。结论 所建立方法可以分析出不同厂家、同厂家不同批次牛黄清心丸(局方)的质量差异,指纹图谱及化学计量分析方法稳定可行,可全面用于其质量评价。

Objective To establish a fingerprint of Niuhuang Qingxin pills(prescription of the bureau)[NQP(Bureau)]and analyze the key components affecting its quality by stoichiometric analysis for providing reference for the quality control and evaluation of NQP(Bureau).Methods The NQP(Bureau)fingerprint was established by high performance liquid chromatography-charged aerosol de-tector(HPLC-CAD)and its methodology was investigated.The common peaks were confirmed and analyzed by cluster analysis,princi-pal component analysis and orthogonal partial least square discriminant analysis.Results RSDs of relative retention time and peak area for precision,stability and repeatability of NQP(Bureau)fingerprint were respectively 0.04%-0.06%and 1.45%-2.67%,0.04%-0.87%and 0.26%-2.87%,0.01%-0.34%and 1.72%-2.93%.Compared with the fingerprint of control samples,26 common peaks were identified,among which 18 characteristic peaks such as amygdalin,glycyrrhizin,baicalin and cholic acid and so on were identified,and the similarity was 0.927-0.995.These samples were divided into three categories by heat map cluster analysis.The cu-mulative contribution rate of the six principal components reached 84.952%.Four compounds causing quality difference,glycyrrhizin,ammonium glycyrrhizinate,coumarin glycoside,and amygdalin,were predicted by orthogonal partial least square discriminant analy-sis.Conclusion The established method can analyze the quality difference of NQP(Bureau)from different manufacturers or diffe-rent batches of the same manufacturer.The fingerprint and stoichiometric analysis are reliable and practicable,and can be comprehen-sively used for the quality evaluation of NQP(Bureau).