基于HPLC指纹图谱、多指标成分含量测定及化学计量学的复方保泰松鸡血藤片质量评价

Quality Evaluation of Compound Phenylbutazone and Spatholobus Stem Tablets Based on HPLC Fingerprint,Determination of Multi-index Components and Chemometrics Analysis

目的建立HPLC指纹图谱和多指标成分定量测定与化学计量学相结合的复方保泰松鸡血藤片质量评价方法。方法以Waters Symmetry C_(18)柱为色谱柱,柱温30℃;乙腈-甲醇(4∶1)与0.05 mol·L^(-1)磷酸二氢钾溶液为流动相,梯度洗脱,流速1.0 mL·min^(-1);检测波长分别为280 nm(检测儿茶素、表儿茶素和美迪紫檀素)、215 nm(检测奇壬醇、豨莶苷和豨莶精醇)和240 nm(检测保泰松、6-姜辣素和8-姜酚)。利用中药色谱指纹图谱相似度评价系统(2012.130723版)建立复方保泰松鸡血藤片HPLC指纹图谱并进行相似度评价;采用SPSS26.0软件聚类分析、主成分分析等化学计量学方法对不同厂家生产的复方保泰松鸡血藤片进行质量评价。结果复方保泰松鸡血藤片HPLC指纹图谱确认了17个共有峰,指认出9个共有峰,3个厂家生产的10批复方保泰松鸡血藤片样品相似度均大于0.95。儿茶素、表儿茶素、美迪紫檀素、奇壬醇、豨莶苷、豨莶精醇、保泰松、6-姜辣素和8-姜酚分别在2.596~259.6、3.148~314.8、0.284 0~28.40、3.876~387.6、0.928 0~92.80、0.512 0~51.20、0.356 0~35.60、2.268~226.8和0.432 0~43.20μg·mL^(-1)内线性关系良好(r≥0.999 1),平均加样回收率分别为99.69%、100.02%、96.91%、100.06%、98.94%、98.42%、97.88%、99.32%和97.32%,RSD分别为0.78%、0.67%、1.24%、0.84%、1.03%、1.36%、1.15%、0.94%和1.46%。10批复方保泰松鸡血藤片样品聚为3类,主成分分析结果显示,主成分1~6是影响复方保泰松鸡血藤片质量评价的主要因子。结论该方法操作简便、重复性好,可用于复方保泰松鸡血藤片的质量控制和综合评价。

OBJECTIVE To establish a quality evaluation method of Compound phenylbutazone and spatholobus stem tablets based on HPLC fingerprint,determination of multi-index components and chemometrics analysis.METHODS The chromatographic column was Waters Symmetry C18 column,and the column temperature was set at 30℃.The mobile phase was composed of acetonitrile-methanol(4∶1)and 0.05 mol·L-1 potassium dihydrogen phosphate solution flowing at 1.0 mL·min-1 in a gradient elution manner.The detection wavelength were set at 280 nm for(+)-catechin,epicatechin and medicarpin,215 nm for kirenol,darutoside and darutigenol,240 nm for phenylbutazone,6-gingerol and 8-gingerol.RESULTS The fingerprint of Compound phenylbutazone and spatholobus stem tablets was established.Seventeen common peaks were confirmed and nine of them were identified.The similarity of fingerprints of 10 batches of Compound phenylbutazone and spatholobus stem tablets from 3 different manufacturers was more than 0.95.(+)-Catechin,epicatechin,medicarpin,kirenol,darutoside,darutigenol,phenylbutazone,6-gingerol and 8-gingerol showed good linear relationships within the ranges of 2.596-259.6,3.148-314.8,0.2840-28.40,3.876-387.6,0.9280-92.80,0.5120-51.20,0.3560-35.60,2.268-226.8 and 0.4320-43.20μg·mL-1(r≥0.9991),whose average recoveries were 99.69%,100.02%,96.91%,100.06%,98.94%,98.42%,97.88%,99.32%and 97.32%with the RSDs of 0.78%,0.67%,1.24%,0.84%,1.03%,1.36%,1.15%,0.94%and 1.46%,respectively.The results of cluster analysis showed that 10 batches of Compound phenylbutazone and spatholobus stem tablets were clustered into three groups,and the results of principal component analysis showed that the principal components 1-6 were the main factor affecting the quality evaluation of Compound phenylbutazone and spatholobus stem tablets.CONCLUSION The method is simple and repeatable,and it can be used to quality control and hensive evaluation of Compound phenylbutazone and spatholobus stem tablets.