不同规格青皮药材UPLC特征图谱和多指标成分含量对比研究

Comparative study on UPLC characteristic chromatogram and multivariate component content of different specifications of Pericarpium Citri Reticulatae Viride

建立个青皮、四花青皮超高效液相色谱(UPLC)特征图谱,结合多成分含量测定,为完善不同规格青皮药材的质量控制提供参考。采用Waters ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.8μm)色谱柱,流动相为乙腈-0.1%甲酸溶液,流速为每分钟0.30 mL,梯度洗脱,检测波长为275 nm,柱温为40℃,进样量为1μL;建立15批个青皮和15批四花青皮的特征图谱,通过对照品比对并结合光谱分析,对共有峰进行鉴定;借助中药色谱指纹图谱相似度评价系统对15批个青皮和15批四花青皮的特征图谱进行相似度评价,通过聚类分析(HCA)、主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)比较两种规格青皮的差异,寻找差异性成分;并对两种规格青皮药材5种有效成分含量进行对比研究。结果显示,个青皮和四花青皮特征图谱均有7个共有峰,指认出其中5个峰,分别为辛弗林、芸香柚皮苷、橙皮苷、川陈皮素和橘皮素;HCA和PCA将青皮药材按规格不同大致分为两类,OPLS-DA发现3个差异性标志物,分别为峰1(辛弗林)、峰4和峰2(橙皮苷)。含量测定研究结果显示,个青皮中辛弗林和橙皮苷的含量高于四花青皮,四花青皮中川陈皮素的含量高于个青皮,差异具有统计学意义(P<0.05)。该方法可以有效鉴别不同规格的青皮药材质量的差异性,为其质量控制提供参考。

UPLC characteristic chromatograms of Fructus Citri Immaturus(FCI)and Pericarpium Citri Reticulatae Viride(PCRV)were established,combined with multi-component content determination,so as to provide reference for the quality control of different specifications of PCRV.Waters ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.8μm)column was used.The mobile phase was acetonitrile-0.1%formic acid solution.The flow rate was 0.30 mL per minute with gradient elution.Detection wavelength was 275 nm.The column temperature was 40°C.The injection volume was 1μL.The characteristic chromatograms of 15 batches of FCI and PCRV were established.Cluster analysis(HCA),principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)were used to compare the differences between the two specifications of PCRV and look for different components.The contents of five effective components in two specifications of PCRV were compared.The result showed that there were seven common peaks in the characteristic chromatograms of FCI and PCRV.Five common peaks were identified,namely synephrine,narirutin,hesperidin,nobiletin and tangeretin by comparing with the reference substances.HCA and PCA roughly divided PCRV into two categories.three differential markers,namely Peak 1(synephrine),peak 4 and peak 2(hesperidin)were found by OPLS-DA.The content determination results showed that the contents of synephrine and hesperidin in Fructus Citri Immaturus were higher than those in PCRV.The contents of nobiletin in PCRV were higher than those in FCI.The differences were statistically significant(P<0.05).This method can effectively analyze the quality differences of different specifications of PCRV and provide reference for the quality evaluation.