HPLC法测定益脑心颗粒中10种成分及化学计量的研究

Determination of 10 components in Yinaoxin Granules by HPLC and stoichiometric research

目的建立测定益脑心颗粒中10种活性成分(天麻素、丹参素钠、对羟基苯甲醇、巴利森苷E、原儿茶醛、巴利森苷B、巴利森苷C、巴利森苷A、二苯乙烯苷、丹酚酸B)含量的高效液相色谱(high performance liquid chromatography,HPLC)法,并结合主成分分析(principal component analysis,PCA)与聚类分析(cluster analysis,CA)对其进行质量评价研究。方法采用Shim-pack Scepter C_(18)(250 mm×4.6 mm,5μm)色谱柱;流动相:乙腈-0.1 g·L^(-1)甲酸水溶液,梯度洗脱;流速:0.8 min·mL^(-1);检测波长:220、310 nm;柱温:20℃;进样量:10μL。采用SPSS 26.0统计软件对15批益脑心颗粒中10种成分的含量进行PCA和CA。结果天麻素、丹参素钠、对羟基苯甲醇、巴利森苷E、原儿茶醛、巴利森苷B、巴利森苷C、巴利森苷A、二苯乙烯苷、丹酚酸B的质量浓度分别在6.99~699.06、11.77~1176.98、6.49~648.76、7.03~702.66、1.77~177.18、1.13~112.50、2.10~210.31、1.30~130.04、6.00~599.76、21.78~2178.54μg·mL^(-1)范围内,线性关系良好(r≥0.9997);10种成分的平均回收率分别为98.43%、97.79%、98.09%、98.53%、98.39%、98.48%、99.05%、97.96%、98.08%、98.27%,RSD值分别为1.36%、1.21%、1.30%、1.53%、1.26%、0.86%、1.17%、1.07%、0.89%、0.92%(n=6)。15批样品聚类分析为两大类,主成分1~4是影响益脑心颗粒质量评价的主要因子,其累积贡献率达87.88%。结论该方法操作简便、稳定可靠、重复性好,可用于益脑心颗粒的质量控制和评价。

Objective To establish a high performance liquid chromatography.(HPLC)method for the simultaneous determination of 10 active components(gastrodin,danshensu sodium p-hydroxybenzyl alcohol,palisenoside E,protocatechuic aldehyde,palisenoside B,palisenoside C,palisenoside A,stilbene glycoside,salvianolic acid B)in Yinaoxin Granules,and the principal component analysis(PCA)and cluster analysis(CA)were used to evaluate its quality.Methods The Shim-pack Scepter C_(18)(250 mm×4.6 mm,5μm)column was used with acetonitrile-0.1 g·L^(-1) formic acid as a mobile phase in gradient elution.The flow rate was 0.8 min·mL^(-1),and the detection wavelength was set at 220 and 310 nm.The column temperature was 20℃,and the injection volume was 10μL.The contents of 10 components in 15 batches of Yinaoxin Granules were analyzed by PCA and CA with SPSS 26.0 statistied software.Results The 10 components showed good resolution.The linear ranges of gastrodin,danshensu sodium,p-hydroxybenzyl alcohol,palisenoside E,protocatechuic aldehyde,palisenoside B,palisenoside C,palisenoside A,stilbene glycoside,and salvianolic acid B were 6.99-699.06,11.77-1176.98,6.49-648.76,7.03-702.66,1.77-177.18,1.13-112.50,2.10-210.31,1.30-130.04 and 6.00-599.76μg·mL^(-1),respectively.The average recoveries of the 10 components were 98.43%,97.79%,98.09%,98.53%,98.39%,98.48%,99.05%,97.96%,and 98.08%,and 98.27%,respectively,with RSD values of 1.36%,1.21%,1.30%,1.53%,1.26%,0.86%,1.17%,1.07%,0.89%,and 0.92%,respectively.15 batches of samples were grouped into 2 categories,and the cumulative contribution rate of the first four principal components was 87.88%.Conclusion The method is simple,stable,reliable and reproducible,which can be used for the quality evaluation of Yinaoxin Granules.