消渴丸HPLC特征图谱研究

目的：建立消渴丸的HPLC特征图谱分析方法。方法：采用高效液相色谱法对20批消渴丸样品进行分析。色谱条件：Welch Materials XB-C₁₈色谱柱（4.6 mm×260 mm，5 μm）；流动相为甲醇-0.02%甲酸水溶液；流速为1.0 mL·min^-1；以葛根素为参照峰，检测波长为280 nm。结果：采用《中药色谱指纹图谱相似度评价系统2.0版》分析了20批消渴丸样品，确定了12个共有峰，20批消渴丸样品的相似度均在0.98以上；利用对照品的保留时间及紫外光谱比较，确认了其中的8个共有峰，分别为3’-羟基葛根素（3号峰）、 葛根素（4号峰）、3’-甲氧基葛根素（5号峰）、大豆苷元-8-C-芹糖基-（1→6）-葡萄糖苷（6号峰）、大豆苷（7号峰）、大豆素（10号峰）、五味子酯甲（11号峰）及五味子甲素（12号峰）。所归属的色谱峰峰面积占总峰面积的80%，占共有峰面积的85%。此外，通过比较消渴丸样品与原药材的HPLC-DAD色谱图，明确了3~7号共有峰来源于葛根，11~12号共有峰来源于南五味子。结论：该方法符合方法学验证要求，可检测消渴丸中的多种成分，所构建的特征图谱能较全面、准确地评价消渴丸质量的均一性，可为消渴丸的整体质量评价提供参考。

Objective: To establish a method for a specific chromatogram analysis of Xiaoke pills by HPLC. Methods: The analysis was performed on a Welch Materials XB-C₁₈ column(4.6 mm×260 mm,5 μm)with a gradient mobile phase of methanol -0.02% formic acid aqueous solution system at a flow rate of 1.0 mL·min^-1.The detection wavelength was 280 nm. Results: The similarity of 20 batch samples was analyzed with Chromatographic Fingerprint Evaluation System Version 2.0 and the overall similarity of all samples exceeded 0.98.Moreover,the specific chromatogram of Xiaokewan was established,and 12 common peaks were found.Among them,8 peaks were identified by comparison with the references or ultraviolet(UV)spectra,including 3'-hydroxy puerarin(peak 3),puerarin(peak 4),3'-methoxy puerarin(peak 5),daidzein-8-C-apiosyl-(1→6)-glucoside(peak 6),daidzin(peak 7),daidzein(peak 10),schisantherin A(peak 11)and deoxyshizandrin(peak 12).The area of identified peaks covered 80 percent of the total peak area,and 85 percent of the area of the 12 common peaks.Besides,the comparison of HPLC-DAD chromatogram between Xiaokewan samples and seven indigenous medicinal materials indicated that five common peaks(peak 3-7)came from Radix Puerariae while peak 11 and peak 12 were from Fructus Schisandrae Sphenantherae.Conclusion: The method conform to the requirements of the methdology validation,and can be used to detect the multiple active ingredients in Xiaokewan.The specific chromatogram is scientific and reasonable as a quality control method for Xiaokewan.