HPLC法测定十味降糖颗粒中10种成分及其化学计量学综合评价  

Determination of 10 components in Shiwei Jiangtang Granules by HPLC and comprehensive evaluation of chemometrics

作　　者：何燕燕 董玉波 徐祥举 张智超 任强[4] 周金辉[4] 王玉娟 HE Yan-yan;DONG Yu-bo;XU Xiang-ju;ZHANG Zhi-chao;REN Qiang;ZHOU Jin-hui;WANG Yu-juan(Department of Pharmacy,No.960 Hospital of PLA Joint Logistics Support Force,Jinan 250031,China;Qilu Pharmaceutical Co.,Ltd.,Jinan 250031,China;Shandong Medical Insurance Center,Jinan 250031,China;School of Pharmacy,Jining Medical University,Rizhao 276826,China)

机构地区：[1]中国人民解放军联勤保障部队第九六〇医院药学部,山东济南250031 [2]齐鲁制药有限公司,山东济南250031 [3]山东省医疗保险事业中心,山东济南250031 [4]济宁医学院药学院,山东日照276826

出　　处：《现代药物与临床》2022年第10期2232-2238,共7页Drugs & Clinic

基　　金：山东省中医药科技发展计划项目(2019-0449)。

摘　　要：目的建立HPLC法同时测定十味降糖颗粒中毛蕊异黄酮葡萄糖苷、芒柄花素、新芒果苷、芒果苷、异芒果苷、3′-羟基葛根素、葛根素、3'-甲氧基葛根素、东莨菪素和东莨菪苷的方法,采用化学计量学对其进行分析。方法采用SepaxBio C18色谱柱(250 mm×4.6 mm,5μm);流动相为乙腈–0.1%甲酸,梯度洗脱;检测波长254 nm(0~45 min测定毛蕊异黄酮葡萄糖苷、芒柄花素、新芒果苷、芒果苷、异芒果苷、3′-羟基葛根素、葛根素、3'-甲氧基葛根素)、346 nm(45~75 min测定东莨菪素和东莨菪苷);柱温30℃;体积流量1.0 mL/min;进样10μL。采用聚类分析、主成分分析、偏最小二乘法–判别分析法对10种成分测定结果进行评价。结果毛蕊异黄酮葡萄糖苷、芒柄花素、新芒果苷、芒果苷、异芒果苷、3′-羟基葛根素、葛根素、3'-甲氧基葛根素、东莨菪素和东莨菪苷分别在1.97~98.50、3.43~171.50、12.35~617.50、6.28~314.00、0.56~28.00、3.99~199.50、15.46~773.00、3.05~152.50、0.43~21.50、0.77~38.50μg/mL有良好的线性关系。平均回收率分别为98.54%、99.63%、100.09%、98.94%、97.11%、99.72%、100.06%、98.71%、96.97%、97.84%,RSD值分别为1.02%、0.84%、0.70%、1.26%、1.36%、0.69%、0.88%、1.42%、1.58%、1.79%。偏最小二乘–判别分析的分类结果与聚类分析的结果基本一致;芒柄花素(VIP=1.575)、东莨菪苷(VIP=1.516)、毛蕊异黄酮葡萄糖苷(VIP=1.223)和葛根素(VIP=1.095)可能是影响十味降糖颗粒产品质量的主要潜在标志物。结论建立的方法准确易行,为十味降糖颗粒多指标成分质量控制方法的制订提供参考。Objective To establish an HPLC method for simultaneous determination of calycosin 7-O-β-D-glucopyranoside,formononetin,neomangiferin,mangiferin,isomangiferin,3'-hydroxy puerarin,puerarin,3'-methoxy puerarin,scopoletin,and scopolin in Shiwei Jiangtang Granules,and to analyze the data by chemometric methods.Methods The separation was carried out on SepaxBio C18 column(250 mm×4.6 mm,5μm).The mobile phase consisted of acetonitrile-0.1%formic acid with gradient elution.The detection wavelengths were set at 254 nm(for calycosin 7-O-β-D-glucopyranoside,formononetin,neomangiferin,mangiferin,isomangiferin,3'-hydroxy puerarin,puerarin,3'-methoxy puerarin)and 346 nm(for scopoletin and scopolin).The column temperature was 30°C,the volume flow was 1.0 mL/min,and the injection volume was 10μL.Cluster analysis,principal component analysis,partial least squaresdiscriminant analysis method were used to evaluate the results of ten components.Results Calycosin 7-O-β-D-glucopyranoside,formononetin,neomangiferin,mangiferin,isomangiferin,3'-hydroxy puerarin,puerarin,3'-methoxy puerarin,scopoletin,and scopolin had good linear relationships within the ranges of 1.97—98.50,3.43—171.50,12.35—617.50,6.28—314.00,0.56—28.00,3.99—199.50,15.46—773.00,3.05—152.50,0.43—21.50,and 0.77—38.50μg/mL.The average recoveries were 98.54%,99.63%,100.09%,98.94%,97.11%,99.72%,100.06%,98.71%,96.97%,and 97.84%with RSD values of 1.02%,0.84%,0.70%,1.26%,1.36%,0.69%,0.88%,1.42%,1.58%,and 1.79%,respectively.The classification results of partial least squares-discriminant analysis were basically consistent with those of cluster analysis.Formononetin(VIP=1.575),scopolin(VIP=1.516),calycosin 7-O-β-Dglucopyranoside(VIP=1.223)and puerarin(VIP=1.095)may be the main potential marker affecting the quality of Shiwei Jiangtang Granules.Conclusion The established HPLC method is convenient and accurate,which can be used for multi-index quality control for Shiwei Jiangtang Granules.

关 键 词：十味降糖颗粒 毛蕊异黄酮葡萄糖苷 芒柄花素 新芒果苷 高效液相色谱 聚类分析 偏最小二乘法–判别分析 

分 类 号：R286.02[医药卫生—中药学]