等效色谱柱选择在大黄和补骨脂多成分高效液相色谱分析中的应用

目的: 尝试将基于保留机制法的反相色谱柱选择策略应用到中药的多成分分析中,并通过实验考察其可行性,为提高法定标准在不同实验室之间的重现性提供依据。方法: 在2台不同的高效液相色谱仪和40根不同品牌、型号的C₁₈色谱柱上,采集大黄、补骨脂中多种成分的液相色谱,对大黄中9种成分、补骨脂中11种成分的分离效果进行评价,尤其是对关键峰对的分析,借助USP法(United States Pharmacopeial approach,USP approach)和PQRI法(PQRI approach)2种等效色谱柱选择方法,比较不同色谱柱对分离的影响。结果: 相似度高的色谱柱对大黄的分离效果接近,对补骨脂的分离效果规律性不明显。结论: 色谱柱对中药成分的分离效果不仅与色谱柱本身的性质有关,与待测成分的性质也相关,因此目前的色谱柱选择系统尚无法解释所有问题。但是相对于随机选择色谱柱,采用这些方法选择色谱柱仍可以提高方法的重现性;结合基于某种中药或某类成分的色谱柱数据库,建立色谱柱选择系统是可行的。

Objective: To provide basis for improving reproducibility of mandatory standards in different laboratories by applying the theory of reversed phase chromatographic column selection based on retention mechanism to analyze traditional Chinese medicine,and validate the feasibility and applicability of this method through experiments. Methods: On 2 different high performance liquid chromatographic instruments with 40 different brands and types of C₁₈ columns,the chromatogram of Rhei Radix et Rhizoma and Psoraleae Fructus was recorded.The separation effect of 9 components in Rhei Radix et Rhizoma and 11 components in Psoraleae Fructus was evaluated,especially the key peaks.The influence of different columns on separation was evaluated by the United States Pharmacopeial approach and the PQRI approach. Results: There was similar separation effect using equivalent columns for Rhei Radix et Rhizoma,but the separation results for Psoraleae Fructus were not regular. Conclusion: The separation effect of chromatographic columns for the ingredients of Chinese medicine was not only related to the properties of the column,but also related to the nature of the constituents,therefore,the current selection system of chromatographic column still could not explain all the problems.However,in contrast to the random selection of chromatographic column,selection of the column using these methods could improve the reproducibility of the analytical method.It is possible to establish the column selection system based on the chromatographic column database of certain traditional Chinese medicine or components.