Szepesi G, Gazdag M, Mihályfi K
Chemical Works of Gedeon Richter, Ltd., Budapest Hungary.
J Chromatogr. 1989 Mar 3;464(2):265-78. doi: 10.1016/s0021-9673(00)94245-6.
The most important steps in the validation of high-performance liquid chromatographic (HPLC) methods are discussed. The establishment of system suitability data and the assessment of peak purity are demonstrated on the example of bisquaternary amino steroids. For the recognition of incomplete resolution of adjacent peak pairs, the absorbance-ratio method in which the ratio of absorbances at two preselected wavelengths are plotted as a function of time in combination with the separation of sample components subjected to various chemical and physico-chemical treatments (stress conditions) is applied. The separation power and performance of the HPLC systems are characterized by the system resolution (SR) and system selectivity (SS). The special demands of stability-indicating methods are summarized.
讨论了高效液相色谱（HPLC）方法验证中最重要的步骤。以双季铵氨基甾体为例，展示了系统适用性数据的建立和峰纯度的评估。为识别相邻峰对的不完全分离，采用吸光度比法，即将两个预选波长处的吸光度之比作为时间的函数进行绘制，并结合对经过各种化学和物理化学处理（强制条件）的样品组分进行分离。HPLC系统的分离能力和性能通过系统分辨率（SR）和系统选择性（SS）来表征。总结了稳定性指示方法的特殊要求。
