[关键词]
[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,5 μm);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10 μL。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、0.77~38.50 μg/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。
[Key word]
[Abstract]
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 C18column (250 mm×4.6 mm,5 μm).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℃,the volume flow was 1.0 mL/min,and the injection volume was 10 μL.Cluster analysis,principal component analysis,partial least squares-discriminant 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 0.77-38.50 μg/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-D-glucopyranoside (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.
[中图分类号]
R286.02
[基金项目]
山东省中医药科技发展计划项目(2019-0449)