基于HPLC指纹图谱结合化学模式识别的黄蛭益肾胶囊质量控制

Quality control of Huangzhi Yishen Capsules based on HPLC fingerprint combined with chemical pattern recognition

目的建立黄蛭益肾胶囊HPLC指纹图谱,采用指纹图谱并结合化学模式识别技术对黄蛭益肾胶囊进行质量控制。方法采用Waters XBridge■Shield RP18(250 mm×4.6 mm,5μm)色谱柱,以乙腈-0.05%磷酸水溶液为流动相,梯度洗脱,体积流量1.0 mL/min,柱温30℃,检测波长210 nm。采用HPLC串联四极杆飞行时间质谱(HPLC-Q-TOF-MS/MS)对共有峰定性分析,采用聚类分析(HCA)、主成分分析(PCA)及偏最小二乘法-判别分析(PLS-DA)统计分析。结果16批样品中共标定24个共有峰,相似度评价均大于0.9,鉴定了其中24个峰并归属于7味药;HCA及PCA识别方法将样品分为3类;PLS-DA筛选出包括黄芪异黄烷苷、人参皂苷Rg_(1)、毛蕊异黄酮葡萄糖苷、7,2’-二羟基-3’,4’-二甲氧基异黄烷、松脂醇二葡萄糖苷、毛蕊异黄酮、芒柄花黄素、绿原酸在内的8种主要差异成分。结论建立的HPLC指纹图谱可最大程度反映黄蛭益肾胶囊的药味组成信息,较全面反映其质量,应用的评价分析技术可合理评价其质量差异并准确寻找出差异成分,为其质量合理控制和标准完善提供依据。

Objective To establish a high performance liquid chromatography(HPLC)fingerprint of Huangzhi Yishen Capsule,control the quality of Huangzhi Yishen Capsule by fingerprint and chemical pattern recognition technology.Methods Waters XBridge■Shield RP18(250 mm×4.6 mm,5μm)column was used,mobile phase was consisted of acetonitrile-0.05% phosphoric acid solution,gradient elution,the flow rate was 1.0 mL/min,the column temperature was 30℃ and the detection wavelength was 210 nm.The common peaks in fingerprint were identified by HPLC-Q-TOF-MS/MS,analyzed by cluster analysis(HCA),principal component analysis(PCA)and partial least squares discriminant analysis(PLS-DA).Results The fingerprint of Huangzhi Yishen Capsule was established with 24 common peaks,the similarities of 16 batches of samples were all above 0.9,24 of them were identified,which come from seven herbs;the samples could be divided into three categories by HCA,PCA and PLS-DA technic,eight components were screened out to cause quality differences,including 2′-hydroxy-3′,4′-dimethoxy-isoflavan-glucoside,Ginsenoside Rg_(1),calycosin-glucopyranoside,7,2′-dihydroxy-3′,4′-dimethoxy-isoflavan,pinoresinol diglucoside,calycosin,formononetin and chlorogenic acid.Conclusion The established HPLC fingerprint can extremely demonstrate the foumlations of Huangzhiyishen Capsule and reflect its quality comprehensively.The application of evaluation software and recognition technics can reasonably evaluate the quality difference and accurately find out the components that caused the quality difference,therefore provide the basis for reasonable quality control and standard improvement on Huangzhi Yishen Capsule.