HPLC多组分定量结合化学计量学及灰色关联度的葆宫止血颗粒质量评价

Quality Evaluation of Baogong Zhixue Granule by HPLC Multi-component Quantitative Combined with Chemometrics and Grey Relational Analysis

目的:建立一种分析葆宫止血颗粒中13个成分的高效液相色谱法,并结合化学计量学及灰色关联度分析对葆宫止血颗粒进行质量评价。方法:采用50%甲醇超声提取样品,CAPCELL PAK C_(18)色谱柱,乙腈-0.2%磷酸水溶液为流动相梯度洗脱,同时测定葆宫止血颗粒中梓醇、地黄苷D、地黄苷A、益母草苷、三七皂苷R_(1)、人参皂苷Rg_(1)、人参皂苷Rb_(1)、柴胡皂苷a、柴胡皂苷d、芍药内酯苷、芍药苷、苯甲酰芍药苷和鹤草酚的含量。采用化学计量学挖掘影响葆宫止血颗粒产品质量的差异标志物,结合灰色关联度法计算各指标的关联度并对13批葆宫止血颗粒的质量进行排序。结果:13个成分的线性范围分别为2.68~134.00、1.64~82.00、0.88~44.00、0.57~28.50、2.35~117.50、11.93~596.50、9.71~485.50、2.05~102.50、1.86~93.00、9.62~481.00、19.05~952.50、1.78~89.00、0.66~33.00µg·mL^(–1)(r>0.999),3个质量浓度水平加样回收率为96.78%~100.12%(RSD<2.0%)。化学计量学分析显示,芍药苷、人参皂苷Rg_(1)、芍药内酯苷、人参皂苷Rb_(1)、梓醇、柴胡皂苷a和三七皂苷R_(1)是影响葆宫止血颗粒产品质量的主要标志物。灰色关联度分析相对关联度为0.3446~0.5742。结论:该方法揭示了影响葆宫止血颗粒质量的主要潜在标志物,综合评价了葆宫止血颗粒的质量,可为其内在整体质量控制提供参考。

Objective:To develop an High Performance Liquid Chromatography(HPLC)method for the simultaneous determination of 13 components in Baogong Zhixue Granule,and to comprehensively evaluate the quality of Baogong Zhixue Granule by chemometrics and grey relational analysis(GRA).Methods:The Baogong Zhixue Granule samples were extracted with 50%methanol solution under ultrasonic condition.The chromatographic separation was performed on a CAPCELL PAK C_(18) column and gradient elution was carried out with acetonitrile-0.2%phosphoric acid aqueous solution as the mobile phase.The contents of catalpol,rehmannioside D,rehmannioside A,leonuride,notoginsenoside R_(1),ginsenoside Rg_(1),ginsenoside Rb_(1),saikosaponin a,saikosaponin d,albiflorin,paeoniflorin,benzoylpaeoniflorin and agrimophol in Baogong Zhixue Granule were determined.Chemometrics was used to explorethe potential markers affecting the product quality of Baogong Zhixue Granule.The relative correlation degree of each index was calculated by GRA method,and the quality of 13 batches of Baogong Zhixue Granule was sorted.Results:The linear ranges of 13 components were 2.68-134.00,1.64-82.00,0.88-44.00,0.57-28.50,2.35-117.50,11.93-596.50,9.71-485.50,2.05-102.50,1.86-93.00,9.62-481.00,19.05-952.50,1.78-89.00 and 0.66-33.00µg·mL^(–1)(r>0.999),respectively,and the average recoveries of three mass concentration ranged from 96.78%-100.12%with RSD<2.0%.Chemometric analysis showed that paeoniflorin,ginsenoside Rg_(1),albiflorin,ginsenoside Rb_(1),catalpol,saikosaponin a and notoginsenoside R_(1) were the main potential markers affecting the quality of Baogong Zhixue Granule.The relative correlation degree was 0.3446-0.5742.Conclusion:This method revealed the main potential markers affecting the quality of Baogong Zhixue Granules,comprehensively evaluated the quality of Baogong Zhixue Granules,and provided a reference for its internal and overall quality control.