双标多测法Ⅰ-双标线性校正技术用于色谱峰的定性

目的:建立一种新的替代对照品模式的多指标含量测定技术——双标多测法,并应用于大黄及其制剂中5种蒽醌的分析。方法: 在5台不同的高效液相色谱仪和14根不同品牌和型号的C₁₈色谱柱上,测定大黄中5种游离蒽醌的峰面积和保留时间,再进行数据分析。定量方面,以芦荟大黄素为参照物,测得与大黄酸、大黄素、大黄酚和大黄素甲醚的相对校正因子。定性方面,以芦荟大黄素和大黄素甲醚2个成分,使用双标线性校正法预测保留时间。结果: 定量方面,相对校正因子可耐受柱温、流速和流动相比例在一定范围内的变动。定性方面,14根色谱柱的验证结果表明,双标线性校正法的预测效果明显优于相对保留时间法。结论: 双标多测法在中药多指标含量测定方面有较好的实用价值和应用前景。

Objective: To establish a new substitute reference substance method named as "two reference substances for determination of multiple components(TRSDMC)" for multi-components assay of rhubarb and its preparations. Methods: The peak area and retention time of the five free anthraquinones in rhubarb were determined on five different HPLC instruments with 14 C₁₈ columns of different brands and types,then the data analysis was conducted.For the quantitative analysis,aloe-emodin was used as the reference substance for determination of relative correction factors of rhein,emodin,chrysophanol and physcion.For the qualitative analysis,both aloe-emodin and physcion were served as linear calibration standards to forecast the retention time of the other three components. Results: In quantitative aspect,the relative correction factors determined were stable in a certain variation range of column temperature,flow rate and proportion of the mobile phase.In qualitative aspect,linear calibration using two reference substances was proved to be more accurate than the relative retention time method for chromatographic peak identification by validation tests with 14 columns. Conclusion: It is demonstrated that TRSDMC is practical and promising for multi-component assay of traditional Chinese medicine.