当归不同药用部位活性成分含量的HPLC分析方法及特征图谱研究

目的：建立同时测定当归不同药用部位中绿原酸、阿魏酸和Z-藁本内酯含量的HPLC分析方法及特征图谱，研究当归不同药用部位成分差异并加以区别。方法：含量测定，采用Agilent ZORBAX SB-C₁₈色谱柱（5 μm，4.6 mm×250 mm），以0.05%乙酸水溶液（A）-甲醇（B）为流动相，梯度洗脱（0~5 min，15%B→40%B；5~14 min，40%B；14~25 min，40%B→65%B；25~35 min，65%B；35~40 min，65%B→90%B；40~45 min，90%B→100%B；45~57 min，100%B），流速0.6 mL·min^-1，采用变波长检测模式（0~30 min，326 nm；30~57 min，350 nm），柱温35 ℃；特征图谱色谱的建立，采用Agilent ZORBAX SB-C₁₈色谱柱（5 μm，4.6 mm×250 mm），以0.05%乙酸水溶液（A）-甲醇（B）为流动相，梯度洗脱（0~5 min，15%B→35%B；5~30 min，35%B→65%B；30~45 min，65%B；45~58 min，65%B→90%B；58~63 min，90%B→100%B；63~75 min，100%B），流速0.6 mL·min^-1，检测波长254 nm，柱温25 ℃。结果：不同产地不同批次当归不同药用部位中绿原酸的含量为0.020%~0.079%，阿魏酸含量为0.025%~0.134%，Z-藁本内酯含量为0.273%~1.46%，有较大差异。特征图谱共确定21个特征峰，对样品特征图谱进行聚类分析、偏最小二乘法分析和主成分分析，样品分为3类。结论：所建立的含量测定方法简单、快速、灵敏、准确，能够用于当归不同药用部位中活性成分的含量测定。所建立的特征图谱方法能较好地反映当归药材的整体情况。两者结合可以更好地对当归药材的质量进行综合评价。当归不同药用部位化合物成分有明显差异，本文研究结果为当归分部位用药提供了一定的理论依据。

Objective: To study the differences in chemical composition among different medicinal parts of Angelicae sinensis radix and distinguish them by determining three active compounds (chlorogenic acid,ferulic acid and Z-ligustilide) and developing characteristic chromatograms using HPLC. Methods: An HPLC method for quantitative analysis was applied with an Agilent ZORBAX SB-C₁₈ column (5 μm,4.6 mm×250 mm) using gradient elution (0-5 min,15%B→40%B;5-14 min,40%B;14-25 min,40%B→65%B;25-35 min,65%B;35-40 min,65%B→90%B;40~45 min,90%B→100%B;45-57 min,100%B) with 0.05% aqueous acetic acid(A)-methanol(B) as the mobile phase at a flow rate of 0.6 mL·min^-1,the compounds were measured at multiple wavelengths (0-30 min,326 nm;30-57 min,350 nm),and the column temperature was set at 35 ℃.For characteristic chromatograms,chromatographic experiments were performed on an Agilent ZORBAX SB-C₁₈ column (5 μm,4.6 mm×250 mm) using gradient elution(0-5 min,15%B→35%B;5-30 min,35%B→65%B;30-45 min,65%B;45-58 min,65%B→90%B;58-63 min,90%B→100%B;63-75 min,100%B) with 0.05% aqueous acetic acid(A)-methanol(B) as the mobile phase at a flow rate of 0.6 mL·min^-1,the detection wavelength was 254 nm,and the column temperature was set at 25 ℃. Results: The contents of chlorogenic acid,ferulic acid and Z-ligustilide ranged 0.020%-0.079%,0.025%-0.134% and 0.273%-1.46% in different medicinal parts of Angelicae sinensis radix from different regions.There were 21 characteristic peaks in the characteristic chromatograms.The samples were divided into three categories by principal component analysis(PCA),partial least squares method(PLS) and cluster analysis. Conclusion: The method for quantitative analysis was sensitive,convenient and practical,and it can be used to determine the active compounds in different medicinal parts of Angelicae sinensis radix.The characteristic chromatograms can be used to evaluate the quality of Angelicae sinensis radix.It was suggested that different medicinal parts of Angelica sinensis varied in the contents of components,which supported the reasonable usage of different parts for medication.