目的 采用高效液相色谱法建立并分析比较含未硫磺熏蒸白芍和硫磺熏蒸白芍的桂枝汤特征图谱。方法 采用Kromasil C18色谱柱（250 mm × 4.6 mm，5 μm），流动相为乙腈-0.1 %甲酸水溶液，梯度洗脱；流速1.0 mL•min-1，柱温30℃，检测波长280 nm，进样量20 μL。结果 通过对13批（5批含未硫熏白芍和8批含硫熏白芍）桂枝汤的HPLC特征图谱研究，共标定了22个特征峰。运用计算机辅助相似性评价系统、聚类分析及主成分分析对所建13批桂枝汤的HPLC特征图谱进行分析比较，结果显示含未硫熏白芍和硫熏白芍的桂枝汤能明显分成两类，且含硫熏白芍的桂枝汤中有新的成分产生。结论 HPLC特征图谱的建立与主成分分析和聚类分析的联合使用是区分含未硫熏白芍和硫熏白芍的桂枝汤的有效方法。该研究为快速鉴别桂枝汤中所用白芍是否为硫熏饮片提供了一定的参考。

Objective To establish and compare HPLC specific chromatograms of Guizhi Decoction containing sun-dried or sulfur-fumigated Radix Paeoniae Alba（RPA）. Methods The separation was performed on a Kromasil C18 column（250 mm × 4.6 mm，5 μm） using a gradient elution with the mobile phase consisting of acetonitrile-0.1 % aqueous formic acid. The flow rate was 1.0 mL•min-1 and the column temperature was 30℃. The detection wavelength was set at 280 nm and 20 μL of the sample was injected for analysis. Results Twenty-two compounds were selected as the characteristic peaks of HPLC specific chromatograms of Guizhi Decoction after analyzing thirteen batches of Guizhi Decoction samples，which included of five batches containing sun-dried RPA and eight batches containing sulfur-fumigated RPA. HPLC specific chromatograms of Guizhi Decoction containing sun-dried RPA or sulfur- fumigated RPA could be easi divided into two categories after investigation with computer-aided similarity evaluation software combined with cluster analysis and principal component analysis. And new components were produced in Guizhi Decoction containing sulfur-fumigated RPA. Conclusion The combined utilization of HPLC specific chromatograms coupled with cluster analysis and principal component analysis is quite effective for the discrimination of Guizhi Decoction containing sun-dried RPA or sulfur-fumigated RPA. The established method provides certain reference for rapid identification of sulfur-fumigated RPA in Guizhi Decoction.

国家自然科学基金资助项目（81173546、30940093）；江苏省自然科学基金资助项目（BK2009495）；江苏省国际科技合作计划资助项目（BZ2011053）；国家药典委员会资助项目；江苏省普通高校研究生科研创新计划资助项目（KYLX_0975）；江苏高校优势学科建设工程资助项目（2011ZYX2-001）。