Multivariate and univariate optimization studies of liquid-chromatographic separation of steroid mixtures.
Open Access
- 1 March 1982
- journal article
- research article
- Published by Oxford University Press (OUP) in Clinical Chemistry
- Vol. 28 (3) , 444-448
- https://doi.org/10.1093/clinchem/28.3.444
Abstract
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.This publication has 0 references indexed in Scilit: