目的: 建立四妙丸指纹图谱,并采用化学计量学方法和双标多测法对四妙丸进行质量评价研究。方法: 采用HPLC法,流动相为乙腈-0.1%磷酸水溶液(梯度洗脱),流速为1 mL·min-1,检测波长为327 nm(0～17 min)、284 nm(18～30 min)、340 nm(31～92 min),柱温为30 ℃。建立四妙丸指纹图谱,使用化学计量学分析进行初步质量评价,再测定四妙丸中8个特征峰在12根不同型号的C18色谱柱上的实际保留时间,双标化合物选定为绿原酸(峰4)和小檗碱(峰14),采用双标线性校正(LCTRS)法预测8个特征峰的保留时间,并在1根未知色谱柱上进行方法验证。再以小檗碱为参照峰,计算绿原酸、黄柏碱的相对校正因子,对各成分进行定量,并将所得结果与外标法进行比较。结果: 13批次四妙丸样品选定16个色谱峰作为指纹图谱的共有峰,经化学计量学分析初步评价其质量;进一步选择采用对照品法归属的8个特征色谱峰进行LCTRS法定性分析。结果表明,LCTRS法预测保留时间的正确率更高(100%),其预测保留时间的准确度优于相对保留时间(RRT)法;绿原酸、黄柏碱相对于小檗碱的相对校正因子分别为0.778 2和0.241 5,以相对校正因子法(RCFM)和常规外标法(ESM)的定量结果显示,二者无显著性差异。结论: 双标线性校正法提高了色谱峰预测保留时间的准确度,能较好地应用于四妙丸多成分的含量测定。
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