逍遥丸的毛细管电泳指纹图谱的建立

采用毛细管区带电泳法建立了逍遥丸(Xiaoyao Pill, XYP)的毛细管电泳指纹图谱(CEFP)。运用正方形优化法,以色谱指纹图谱分离量指数(RF)为优化的目标函数,对建立指纹图谱的实验条件进行了优化,确定了最佳背景电解质(BGE)溶液50 mmol/L硼砂-50 mmol/L磷酸氢二钠-150 mmol/L磷酸二氢钠-50 mmol/L碳酸氢钠(1:1:1:5, v/v/v/v; pH 7.40)、紫外检测波长228 nm、运行电压12 kV、重力进样25 s (高度14 cm)的分离检测条件。采用未涂层石英毛细管(70 cm×75 μm,有效分离长度57 cm)分离,以咖啡酸色谱峰为参照,确定13批逍遥丸样品的21个共有指纹峰。通过聚类分析确定用其中10批样品生成对照CEFP,以此为标准用系统指纹定量法鉴别13批逍遥丸的质量,结果显示: S3号样品的化学成分数量和分布比例不合格,S10和S12号样品含量明显偏高,其余批次质量均合格。所建立的正方形优化法操作简便,适用于中药的毛细管区带电泳BGE的选择;所建立的逍遥丸CEFP具有较好的精密度和重现性,可以为逍遥丸的质量控制提供新的参考。

Establishment of capillary electrophoresis fingerprints of Xiaoyao pills

The capillary electrophoresis fingerprint (CEFP) of Xiaoyao Pills (XYP) was established by capillary zone electrophoresis (CZE). The quadrilateral method was developed for the optimization of the background electrolyte (BGE), and the running voltage, the compositions and pH of BGE, and so on, were evaluated by the chromatographic fingerprint resolution index (RF). An uncoated fused silica capillary column (70 cm x 75 microm, the effective length of 57 cm) was employed; 50 mmol/L sodium borate-50 mmol/L Na2HPO4-150 mmol/L NaH,PO,-50 mmol/L NaHCO3 (1: 1:1:5, v/v/v/v, adjusted to pH 7.40) was used as the ideal BGE. The detection wavelength was set at 228 nm, and a voltage of 12 kV and hydrodynamic pressure injection in 25 s at the height of 14 cm were applied. The CEFPs from 13 batches of XYPs containing 21 common peaks were obtained with good precision and reproducibility, in which caffeic acid was selected as the reference peak. Cluster analysis indicated that 10 out of 13 batches of XYP samples can be deemed as the reference CEFP (RCEFP). The whole 13 batches of XYP were evaluated by the systematic quantified fingerprint method (SQFM) with the RCEEP as the qualified model. The results indicated that the contents of S10 and S12 were obviously higher, the chemical constituents quantity and distributed proportion of S3 were not qualified, while the others were completely qualified. It is concluded that the method can be applied to the optimization of BGE in CZE in traditional Chinese medicine separation; and the established CEFPs of XYP can be served as a novel reference to identify and control the quality of XYP.