热痹痛合剂HPLC指纹图谱的建立和芍药苷、柚皮苷、黄芩苷、川续断皂苷Ⅵ的测定  

Establishment of HPLC fingerprint of Rebitong Mixture and determination of paeoniflorin,naringin,baicalin,and asperosaponinⅥ

作　　者：邱琼华 李隽永 韦倩 毛桂福 QIU Qiong-hua;LI Juan-yong;WEI Qian;MAO Gui-fu(Liuzhou Traditional Chinese Medical Hospital(Liujcouh Si Ywcuengh YihYen),Liuzhou 545001,China)

机构地区：[1]柳州市中医医院(柳州市壮医医院),广西柳州545001

出　　处：《现代药物与临床》2022年第4期737-745,共9页Drugs & Clinic

基　　金：广西中医药壮瑶医药院内制剂孵化基地项目(桂中医药发[2021]7号)。

摘　　要：目的建立热痹痛合剂的HPLC指纹图谱和热痹痛合剂中芍药苷、柚皮苷、黄芩苷、川续断皂苷VI的HPLC测定方法。方法指纹图谱分析采用Agilent 1260 C18色谱柱(250 mm×4.6 mm,5μm),以乙腈–0.2%磷酸溶液为流动相,梯度洗脱;检测波长为212 nm;体积流量为1 mL/min;柱温30℃;进样量10μL。对指纹图谱进行相似度评价,并运用聚类分析、主成分分析、正交偏最小二乘法–判别分析分析热痹痛合剂样品图谱。采用HPLC法测定芍药苷、柚皮苷、黄芩苷、川续断皂苷VI,采用Agilent 1260 C18色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈–0.2%磷酸溶液,梯度洗脱;检测波长230 nm(芍药苷)、280 nm(柚皮苷、黄芩苷)、260 nm(川续断皂苷VI);体积流量1 mL/min;柱温30℃;进样量为10μL。结果10批热痹痛合剂样品的指纹图谱中有26个共有峰,相似度均大于0.99,化学模式识别将10批样品分为2大类,OPLS-DA分析筛选出4个对制剂质量差异影响较大的化合物。含量测定中芍药苷、柚皮苷、黄芩苷、川续断皂苷VI分别在27.238~272.384、38.207~382.066、77.864~778.642、51.015~510.015μg/mL线性良好,平均回收率分别为95.34%、103.51%、103.51%、102.68%,RSD值分别0.65%、1.12%、0.86%、2.41%。结论所建立的方法准确、合理,重复性好,可用于热痹痛合剂的质量控制和评价。Objective To establish the HPLC fingerprint of Rebitong Mixture and to simultaneously determine the contents of paeoniflorin,naringin,baicalin,and asperosaponinⅥin Rebitong Mixture by HPLC method.Methods The fingerprint analysis was adopted on Agilent 1260 C18(250 mm×4.6mm,5μm)with the mobile phase consisted of acetonitrile-0.2%phosphoric acid flowing at 1.0 mL/min in gradient elution manner,and the detection wavelength was set at 212 nm with column temperature 30℃.Similarity of fingerprints was evaluated,and cluster analysis(CA),principal component analysis(PCA)and orthogonal partial least squaresdiscriminant analysis(OPLS-DA)were used to analyze the sample patterns.The contents of paeoniflorin,naringin,baicalin,and asperosaponinⅥin Rebitong Mixture were determined by HPLC method on Agilent 1260 C18 column(250 mm×4.6 mm,5μm).Acetonitrile-0.2%phosphoric acid was used as the mobile phase by gradient elution.The detection wavelength were 230 nm for paeoniflorin,280 nm for naringin and baicalin,and 260 nm for asperosaponinⅥ.The flow rate was 1 mL/min,the column temperature was 30℃,and the injection volume was 10μL.Results There were 26 common peaks in the fingerprints of 10 batches of Rebitong Mixture samples,and the similarity was all above 0.99.The 10 batches of samples were divided into 2 categories with chemical pattern recognition,and four peaks that had a greater impact on the quality of preparations were selected by OPLS-DA analysis.The linearity of paeoniflorin,naringin,baicalin,and asperosaponinⅥwas 27.238—272.384,38.207—382.066,77.864—778.642,51.015—510.015μg/mL,and the average recoveries were 95.34%,103.51%,103.51%,and 102.68%with RSD values of 0.65%,1.12%,0.86%,and 2.41%,respectively.Conclusion The method is accurate,reasonable,and reproducible,and can be used for the quality control and evaluation of Rebitong Mixture.

关 键 词：热痹痛合剂 指纹图谱 相识度评价 聚类分析 芍药苷 柚皮苷 黄芩苷 川续断皂苷VI 高效液相色谱 

分 类 号：R286.02[医药卫生—中药学]