HPLC-PDA-ELSD测定参芪五味子胶囊中10个成分含量及化学模式识别研究

Simultaneous Determination of Ten Components in Shenqi Wuweizi Capsule by HPLC-PAD-ELSD Combined with Chemical Pattern Recognition

关键词:  参芪五味子胶囊  高效液相色谱法  含量测定  模式识别  主成分分析

Key words:  Shenqi Wuweizi capsule  HPLC  content determination  pattern recognition  principal component analysis (PCA)

摘要:

目的 建立HPLC-PDA-ELSD同时测定参芪五味子胶囊中毛蕊异黄酮葡萄糖苷、芒柄花苷、酸枣仁皂苷A、酸枣仁皂苷B、黄芪甲苷、党参炔苷、斯皮诺素、五味子酯甲、五味子酯乙、五味子甲素10个有效成分含量的方法，并结合主成分分析对其进行质量评价研究。方法 样品采用70%甲醇提取后，采用Waters Symmetry C¹⁸色谱柱（4.6 mm×250 mm，5 μm）分离，流动相为含5%四氢呋喃的乙腈（A）-0.05%甲酸水溶液（B），梯度洗脱；流速为1.0 mL·min^-1；柱温35℃；进样量10 μL；毛蕊异黄酮葡萄糖苷、芒柄花苷、党参炔苷、斯皮诺素、五味子酯甲、五味子酯乙、五味子甲素检测波长：230 nm；酸枣仁皂苷A、酸枣仁皂苷B、黄芪甲苷使用ELSD检测器，漂移管温度60℃。并采用SIMCA14.1统计软件对含量测定结果进行主成分分析。结果 在一定浓度范围内，参芪五味子胶囊中10个成分线性关系良好（r>0.999）；平均加样回收率（n=9）为98.28%~102.95%，RSD均<2.0%。15批样品中毛蕊异黄酮葡萄糖苷、芒柄花苷、酸枣仁皂苷A、酸枣仁皂苷B、黄芪甲苷、党参炔苷、斯皮诺素、五味子酯甲、五味子酯乙、五味子甲素含量分别为0.136~0.152，0.296~0.323，0.334~0.403，0.074~0.114，2.401~2.504，1.455~1.512，0.094~0.164，2.031~2.214，0.711~0.809，1.892~1.973 mg·g^-1。主成分分析将参芪五味子胶囊分类为3类。结论 所建立的HPLC-PDA-ELSD测定参芪五味子胶囊中10个成分含量方法快捷、准确、重复性好，专属性强，结合主成分分析法可为参芪五味子胶囊的质量控制提供参考。

Abstract:

OBJECTIVE To develop an HPLC-PDA-ELSD method for the simultaneous determination of calycosin7-O-β- D-Glucopyranoside, ononin, jujuboside A, jujuboside B, astragaloside IV, lobetyolin, spinosin, schisantherin A, schisantherin B, deoxyschizandrin in Shenqi Wuweizi capsule, and to evaluate the quality of Shenqi Wuweizi capsule combined with principal component analysis. METHODS After the sample was extracted with 70% methanol, it was separated on a Waters Symmetry C₁₈ column(4.6 mm×250 mm, 5 μm), and the mobile phase was acetonitrile(A) containing 5% tetrahydrofuran-0.05% formic acid aqueous solution(B) with gradient elution, the flow rate was 1.0 mL·min^-1, and the column temperature was 35℃, the sample volume was 10 μL. Detection wavelength for calycosin7-O-β-D-glucopyranoside, ononin, lobetyolin, spinosin, schisantherin A, schisantherin B, deoxyschizandrin were set at 230 nm and ELSD was used for detection of jujuboside A, jujuboside B and astragaloside IV. The temperature of the drift tube was 60℃. The principal component analysis were conducted for the results of content determination by SIMCA14.1 statistical software. RESULTS Within a certain concentration range, the ten ingredients all showed good linear relationship(r>0.999), and the average recoveries were 98.28%^-102.95% and RSDs were <2.0%. The contents of calycosin7-O-β-D-glucopyranoside, ononin, jujuboside A, jujuboside B, astragaloside IV, lobetyolin, spinosin, schisantherin A, schisantherin B, deoxyschizandrin in 15 samples were 0.136-0.152, 0.296-0.323, 0.334-0.403, 0.074-0.114, 2.401-2.504, 1.455^-1.512, 0.094-0.164, 2.031-2.214, 0.711-0.809, 1.892^-1.973 mg·g^-1, respectively. The samples could be clearly classified into three categories by using PCA. CONCLUSION The established HPLC-PDA-ELSD method for the determination of 10 components in Shenqi Wuweizi capsule is fast, accurate, reproducible and specific, which can provide a reference for the quality control of Shenqi Wuweizi capsule combined with principal component analysis.