摘要
建立盐杜仲的HPLC指纹图谱及多成分含量测定方法,结合指纹图谱相似度评价、聚类分析(cluster analysis,CA)、主成分分析(principal component analysis,PCA)和正交偏最小二乘法判别分析(orthogonal partial least square discriminate analysis,OPLS-DA)进行不同来源盐杜仲的质量评价.采用Cosmosil 5C18-MS-Ⅱ色谱柱(4.6 mm×250 mm,5 μm),以甲醇-0.2%磷酸水溶液为流动相,梯度洗脱,流速1.0 mL·min-1,检测波长238 nm,柱温 30℃,进样量 10 μL.20 批不同来源盐杜仲的指纹图谱相似度评价结果表明,除S3批次样品指纹图谱相似度为0.893外,其余19批次样品的相似度均≥0.919,表明相似度较好;共标定 14个共有峰,并指认了京尼平苷酸等7个共有峰,京尼平苷酸、绿原酸、京尼平苷、京尼平、松脂醇二葡萄糖苷、鹅掌楸苷和松脂醇单葡萄糖苷7个成分的质量分数依次为 0.062 0%～0.426 9%、0.024 9%～0.116 5%、0.009 5%～0.052 9%、0.005 5%～0.034 8%、0.115 9%～0.317 8%、0.016 4%～0.108 8%、0.026 4%～0.039 8%;通过CA、PCA和OPLS-DA均可将20批盐杜仲划分为3类,且通过OPLS-DA项下的变量重要性投影(variable importance in projection,VIP)值分析筛选出京尼平苷酸和绿原酸2个差异质量标志物.该研究建立的HPLC指纹图谱和多成分含量测定方法稳定、可靠,可为盐杜仲的质量控制提供参考.
Abstract
This study established an HPLC fingerprint and multi-component content determination method for salt-fired Eucommiae Cortex,and evaluated the quality of salt-fired Eucommiae Cortex from different sources using fingerprint similarity evaluation,cluster analysis(CA),principal component analysis(PCA),and orthogonal partial least square discriminate analysis(OPLS-DA).HPLC was launched on a Cosmosil 5C18-MS-Ⅱ column(4.6 mm×250 mm,5 μm)by gradient elution with a mobile phase of methanol-0.2%phosphoric acid aqueous solution at a flow rate of 1.0 mL·min-1,detection wavelength of 238 nm,column temperature of 30℃,and an injection volume of 10 μL.The results of fingerprint similarity evaluation for 20 batches of salt-fired Eucommiae Cortex indicated that,except for batch S3 with a similarity of 0.893,the similarity of the other 19 batches was of≥0.919,suggesting good similarity.Fourteen common peaks were calibrated and seven common peaks were identified including geniposidic acid.The mass fractions of ge-niposidic acid,chlorogenic acid,geniposide,genipin,pinoresinol diglucoside,liriodendrin,and pinoresinol-4-O-β-D-glucopyranoside were 0.062 0%-0.426 9%,0.024 9%-0.116 5%,0.009 5%-0.052 9%,0.005 5%-0.034 8%,0.115 9%-0.317 8%,0.016 4%-0.108 8%,and 0.026 4%-0.039 8%,respectively.Using CA,PCA,and OPLS-DA,the 20 batches of salt-fired Eucommiae Cortex were classified into three categories.Additionally,through the analysis of variable importance in projection(VIP)under OPLS-DA,two differential quality markers,geniposidic acid and chlorogenic acid,were identified.The established HPLC fingerprint and multi-component content determination method is stable and reliable,providing a reference for quality control of salt-fired Eucommiae Cortex.
NETL
NSTL
维普
万方数据