芪苓益肾颗粒高效液相色谱指纹图谱物质基准研究及7种成分含量测定

HPLC fingerprint of Qiling Yishen granules and the determination of 7 components

目的建立芪苓益肾颗粒物质基准的高效液相色谱指纹图谱,并建立同时测定其7种指标成分含量的方法。方法采用GL Sciences ODS-3(5μm,4.6 mm×250 mm)色谱柱,0.05%磷酸-水-乙腈为流动相梯度洗脱,流速1 mL·min^(-1),柱温35℃,检测波长254 nm。通过相似度分析,对共有峰进行归属。采用聚类分析,主成分分析及正交偏最小二乘法-判别分析对指纹图谱数据进行评价,并筛选关键性成分,测定7种活性成分的含量。结果15批芪苓益肾颗粒物质基准的对照指纹图谱的相似度均超过0.93,标定了22个共有峰,指认了18个色谱峰。15批物质基准分为两类,并筛选出5种关键性成分,分别是峰10(毛蕊异黄酮葡萄糖苷)、峰17(芒柄花苷)、峰18(毛蕊异黄酮)、峰1及峰20(芒柄花素)。同时测定了毛蕊异黄酮葡萄糖苷、阿魏酸、洋川芎内酯Ⅰ、芒柄花苷、毛蕊异黄酮、芒柄花素、藁本内酯共7种成分的含量,各物质基准的质量分数范围分别是1.09~1.68、0.27~0.63、0.06~0.45、0.51~0.99、0.31~0.68、0.29~0.59、0.25~0.96 mg·g^(-1)。结论该方法操作简单、重复性、稳定性较好,可为该制剂物质基准的质量控制与评价提供参考。

Objective To establish a high performance liquid chromatography (HPLC) fingerprint for the Qiling Yishen granules substance standard,and to simultaneously determine the contents of7 index components.Methods GL Sciences ODS-3 (5μm,250 mm×4.6 mm) chromatographic column was used with 0.05%phosphoric acid-water (A)-acetonitrile (B) as the mobile phase in gradient elution.The flow rate was 1 mL·min^(-1),the column temperature was 35℃,and the detection wavelength was 254 nm.The total peak was assigned by similarity analysis.Cluster analysis (CA),principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used to evaluate the fingerprint data and screen the key components.The contents of 7 active ingredients was determined.Results The similarity of the control fingerprint spectrum based on the Qiling Yishen granule.Totally 22 consensus peaks were standardized,and 18columns peaks were identified.CA,PCA and OPLS-DA divided the 15 batches of material standards into two categories.Five key components were screened out,namely peak 10 (calycosin 7-O-β-D-glucopyranoside),peak 17 (formononetin),peak 18 (calycosin),peak 1 and peak 20 (formononetin).The contents of 7 components including verbasil glucoside,ferulic acid,ligustrideⅠ,formononetin,calycosin,formononetin,and ligustilide were at 1.09-1.68,0.27-0.63,0.06-0.45,0.51-0.99,0.31-0.68,0.29-0.59,0.25-0.96 mg·g^(-1).Conclusion The method is simple,reproducible,and stable,which can provide reference for the quality control and evaluation of the preparation material standard.