双标多测法Ⅱ—检测波长选择对定量分析的影响

目的: 为解决替代对照品法或一测多评法在相对校正因子法定量和相对保留时间法定性方面的误差有时偏大的问题,建立了一种新的替代对照品模式的多指标含量测定法—双标多测法。 方法: 在3台不同品牌的高效液相色谱仪和23种不同品牌和型号的C₁₈色谱柱上,测定重楼中4个皂苷类成分的峰面积和保留时间,计算相应的相对校正因子和相对保留时间,再进行数据偏差的比较和分析。 结果: 定量方面,解释了检测波长和仪器波长偏移影响相对校正因子的机制,并据此提出了波长选择建议;定性方面,证明了梯度条件下,双标线性校正法预测保留时间的效果仍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具有较好的实用价值和较广的应用前景。

Objective: To establish a new pattern of substitute reference substance method,namely two reference substances for determination of multiple components in order to decrease the deviation of relative correction factor method for quantitative analysis and relative retention time method for peak identification. Methods: The peak area and the retention time were recorded for the four kinds of Chonglou saponin in Paridis Rhizoma on three different brands of HPLC instruments with 23 different brands C₁₈ columns,then their relative correction factors and relative retention times were calculated for comparison of deviations. Results: In the quantitative aspect,the mechanism of detection wavelength and instrument wavelength shift influencing relative correction factor measurement was explained,and accordingly advice on wavelength selection was given.In the qualitative aspect,linear calibration using two reference substances was proved to be better than relative retention time method under gradient elution condition. Conclusion: It is demonstrated that the method of two reference substances for determination of multiple components is suitable and practical for quality evaluation of traditional Chinese medicine.