[关键词]
[摘要]
目的 建立芪白平肺颗粒(QPG)的HPLC指纹图谱,并进行多成分定量分析,用于评价QPG的质量。方法 样品经50%甲醇提取后,采用Phenomenex Luna C18柱(250 mm×4.6 mm,5 μm)进行检测,以甲醇-0.2%甲酸水溶液梯度洗脱,体积流量1.0 mL/min,检测波长250 nm,柱温30℃。采用国家药典委员会出版的《中药色谱指纹图谱相似度评价系统》(2012年版),对10批次的QPG化学成分指纹图谱进行相似度计算,并通过对照品对共有峰进行指认。结果 10批QPG指纹图谱中样品间相似度均大于0.96,共标定出25个共有峰,各峰分离度良好,其中1号峰来源于地龙,2、3号峰来源于地龙和人参,4号峰来源于人参、五味子和川芎,5号峰来源于人参、薤白、川芎和地龙,6、8、22、23、24、25号峰来源于五味子,7号峰来源于黄芪和五味子,9号峰来源于人参和薤白,10号峰来源于薤白和葶苈子,11、12、13、15、16号峰来源于川芎,14号峰来源于黄芪和葶苈子,17、18、19、20、21号峰来源于黄芪。通过对照品比对确定了6个成分,分别为咖啡酸(12号峰)、阿魏酸(13号峰)、五味子醇甲(22号峰)、五味子酯甲(23号峰)、五味子甲素(24号峰)和五味子乙素(25号峰);其中,五味子醇甲在2.99~95.61 μg/mL、咖啡酸在3.38~108.02 μg/mL、阿魏酸在3.60~115.33 μg/mL、五味子甲素在3.26~104.17 μg/mL、五味子乙素在2.89~92.45 μg/mL、五味子酯甲在2.81~89.77 μg/mL,呈良好的线性关系;10批QPG样品中咖啡酸在0.412~0.429 mg/g,阿魏酸在0.302~0.317 mg/g,五味子醇甲在0.182~0.195 mg/g,五味子酯甲在0.179~0.195 mg/g,五味子甲素在0.203~0.215 mg/g,五味子乙素在0.131~0.144 mg/g,不同批次间各指标成分的量变化较小,样品质量较稳定。结论 本方法具有良好的精密度、重复性、稳定性,色谱峰间分离度较好,可用于QPG的质量评价。
[Key word]
[Abstract]
Objective To establish an HPLC fingerprint of the compounds in Qibai Pingfei Granules (QPG), and to make a quantitative analysis. Methods Sample was extracted by 50% methanol. Phenomenex Luna C18 column (250 mm×4.6 mm, 5 μm) was used with a mobile phase of methanol-0.2% formic acid gradient elution. The flow rate was 1.0 mL/min, the detection wavelength was 250 nm, and the column temperature was 30℃. The chemical component fingerprint similarity of 10 batches of QPG was calculated with Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012) published by National Pharmacopoeia Committee and the common peaks were identified by reference compounds. Results Fingerprints of 10 batches of QPG were established and the similarities to the common mode were above 0.96. Totally 25 common peaks were found. Among them, peak 1 belonged to Pheretima, peaks 2 and 3 belonged to Pheretima and Ginseng Radix et Rhizoma (GRR), peak 4 belonged to GRR, Schisandrae Chinensis Fructus (SCF), and Chuanxiong Rhizoma (CR), peak 5 belonged to GRR, Allii Macrostemonis Bulbus (AMB), CR, and Pheretima, peaks 6, 8, 22, 23, 24, and 25 belonged to SCF, peak 7 belonged to Astragali Radix (AR) and SCF, peak 9 belonged to GRR and AMB, peak 10 belonged to Descurainiae Semen Lepidii Semen (DSLS) and AMB, peaks 11, 12, 13, 15, and 16 belonged to CR, peak 14 belonged to AR and DSLS, peaks 17, 18, 19, 20, and 21 belonged to AR. Based on the retention time, and UV absorption spectra of reference compounds, six constituents including caffeic acid (peak 12), ferulic acid (peak 13), schizandrol A (peak 22), schisantherin A (peak 23), deoxyschizandrin (peak 24), and schisandrin (peak 25) were identified. The linear ranges of caffeic acid, ferulic acid, schizandrol A, schisantherin A, deoxyschizandrin, and schisandrin were 3.38-108.02, 3.60-115.33, 2.99-95.61, 2.81-89.77, 3.26-104.17, and 2.89-92.45 μg/mL, respectively. In 10 batches of QPG samples, the contents were as follows:caffeic acid of 0.412-0.429 mg/g, ferulic acid of 0.302-0.317 mg/g, schizandrol of A 0.182-0.195 mg/g, schisantherin A of 0.179-0.195 mg/g, deoxyschizandrin of 0.203-0.215 mg/g, and schisandrin of 0.131-0.144 mg/g, the amount of each indicator composition among different batches changed a litte, and the sample quality is stable. Conclusion The method has good precision, reproducibile, stability, separation, and can be used for the quality control of QPG.
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[基金项目]
现代中药创新集群与数字制药技术平台(2013ZX09402203)