甾类孕激素的高效液相色谱分离流动相选择最优化法

本文就近年来发展的几种HPLC流动相选择最优化理论进行系统研究并加以综合应用。文内对醋酸甲地孕酮等六种常用甾类孕激素混合组分的分离,初步建立了经过梯度洗脱法初选流动相、等洗脱强度二元流动相转换及序贯单纯形寻找三元最佳流动相等三个实验步骤的HPLC最优化法,可迅速、有效地获得使混合组分在给定的分析时间内(1≤K≤10)达到最大分离度的最优流动相组成:MeOH—THF—H₂O(41:22:37)及MeOH—ACN—H₂O(61.6:8.9:29.5)。

OPTIMIZATION OF HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC SEPARATION FOR PROGESTOGENS

An efficient, rapid HPLC optimization method based on semiempirical model and simplex algorithm was proposed and examplified by the optimization separation of six progestogens within a given time (1≤K≤10). The resultant composition of optimum ternary mobile phase was MeOH—THF—H₂O (41:22:37), MeOH—ACN—H₂O (61.6:8.9:29.5) respectively.The procedure consists of three main steps: 1. Initial estimation of isocratic condition using MeOH—H₂O linear gradient run. 2. Transfer MeOH—H₂O to other iso-eluotropic binary mobile phases. 3. Simplex algorithm. In step three, a small step size simplex was used instead of "large step size simplex". By combining the binary conditions obtained from step 1 and 2, the iso-eluotropic ternary mobile phase composition was made as initial simplex points, which were closer to the optimum regions than that of "large step size" because many failed experiments caused by boundary violation and analysis time excess were avoided.A computer program in BASIC was presented for "off line" optimization of HPLC using the simplex method.