藏药方剂十味诃子丸HPLC指纹图谱分析及4种成分含量测定

HPLC fingerprint analysis and content determination of four components of Tibetan medicine prescription Shiweihezi Pills

目的:建立藏药方剂十味诃子丸的HPLC指纹图谱及含量测定方法。方法:采用Swell Chromplus?C18(4.6mm×250 mm,5μm)色谱柱检测分析,以甲醇-0.2%的磷酸水为流动相进行梯度洗脱,进样量10μl,流速0.8 ml/min,柱温30℃,检测波长235 nm,对同一厂家6个批次十味诃子丸样品进样分析。结果:建立了藏药方剂十味诃子丸的HPLC指纹图谱,确定了23个共有峰,对共有峰进行了制剂-药材的来源归属和指认。使用《中药色谱指纹图谱相似度评价系统2004A版》计算相似度,与对照指纹图谱的相似度均>98%。采用SPSS 19.0和SIMCA 14.1软件并结合聚类分析(CA)、主成分分析(PCA)与正交偏最小二乘法-判别分析(OPLS-DA)对23个共有峰面积结果进行分析,3种化学模式识别方法均可将样品分为2类,生产年份是造成批次S1与S2-S6差异的主要原因。筛选出了13种主要差异成分,其中已指认出的为柯里拉京和鞣花酸,结合复方来源归属和指认的君药成分没食子酸和芒果苷,对这4种有效成分进行含量测定,其含量测定结果显示,4号峰没食子酸、13号峰柯里拉京、15号峰芒果苷、21号峰鞣花酸的含量分别为8.900~15.891、0.535~1.984、0.292~0.342、23.611~31.739 mg·g^(-1)。结论:该研究建立的藏药方剂十味诃子丸HPLC指纹图谱及多成分含量测定的方法操作简单,重复性较好,稳定、可靠,可为藏药十味诃子丸的质量评价提高提供参考。

Objective:To establish the HPLC fingerprint and content determination method of Tibetan medicine prescription Shiweihezi Pills.Method:The Swell Chromplus?C18(4.6 mm×250 mm,5μm)chromatographic column was used for detection and analysis,with methanol-0.2%phosphoric acid water as the mobile phase gradient elution,at a flow rate of 0.8 mL·min^(-1)with column temperature was 30℃.The injection volume was 10μL and the detection wavelength was 235nm.The samples of 6 batches of Shiweihezi pills from the same manufacturer were analyzed.Result:The HPLC fingerprint of Tibetan medicine prescription Shiweihezi Pills was established,and 23 common peaks were identified.Using the"Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System Version 2004A"to calculate the similarity,the similarity between all samples and the control fingerprints was>98%.Using SPSS 19.0 and SIMCA 14.1 software combined with cluster analysis(CA),principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)to analyze the results of 23 common peak areas,3 chemical pattern recognition method can classify the samples into two categories,and the production year was the main reason for the difference between batches S1 and S2-S6.13 main differential components were screened out,among which the identified components were corilagin and ellagic acid.Combined with the source of the compound and the identified monarch medicinal components gallic acid and mangiferin,the contents of the four active ingredients were determined.The contents of gallic acid(Peak 4),corragine(Peak 13),mangiferin(Peak15),ellagic acid(Peak 21)were 8.900~15.891,0.535~1.984,0.292~0.342,23.611~31.739 mg·g^(-1),respectively.Conclusion:The method established in this study for the HPLC fingerprint of Tibetan medicine Shiweihezi Pills and the determination of the multi-component content is simple to operate,has good repeatability,is stable and reliable,and can provide a reference for the improvement of the quality evaluation of Tibetan medicine Shiweihezi Pills.