Fast D M, Culbreth P H, Sampson E J
Clin Chem. 1982 Mar;28(3):444-8.
We developed a high-performance liquid-chromatographic separation of five steroids (estriol, estradiol, cortisol, progesterone, and testosterone), eluting with a water-acetonitrile gradient from a reversed-phase (C18) column. By applying a simplex search algorithm to maximize a chromatographic-response function, we sought to optimize the original conditions of the chromatographic analysis, which did not separate two pairs of overlapping peaks. Our chromatographic-response function incorporated both peak separation and total time of analysis. Three factors were varied simultaneously to maximize this function: flow rate, column temperature, and gradient shape. From the simplex optimization, we selected a flow rate of 1.50 mL/min, a temperature of 52 degrees C, and a linear gradient for our analysis. Subsequent univariate studies of the initial mobile phase composition showed that acetonitrile/water (20/80 by vol) gave an adequate separation.
我们开发了一种高效液相色谱法来分离五种甾体激素（雌三醇、雌二醇、皮质醇、孕酮和睾酮），使用水 - 乙腈梯度从反相（C18）柱上洗脱。通过应用单纯形搜索算法来最大化色谱响应函数，我们试图优化色谱分析的原始条件，原始条件下两对峰存在重叠未分开。我们的色谱响应函数同时考虑了峰分离度和总分析时间。同时改变三个因素以最大化该函数：流速、柱温以及梯度形状。通过单纯形优化，我们选择了1.50 mL/min的流速、52℃的温度以及线性梯度用于我们的分析。随后对初始流动相组成的单变量研究表明，乙腈/水（体积比20/80）能实现充分分离。
