The influence of temperature on the multiple separation of estrogenic steroids using mobile phases modified with β-cyclodextrin in high-performance liquid chromatography

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 15; no. 9; pp. 1281 - 1287
• Main Authors: Zarzycki, P.K., Wierzbowska, M., Lamparczyk, H.
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Amsterdam Elsevier B.V 01-06-1997; Elsevier Science
• Subjects: Analytical, structural and metabolic biochemistry; beta-Cyclodextrins; Biological and medical sciences; Chromatography, High Pressure Liquid; Cyclodextrin; Cyclodextrins; Estrogens; Estrogens - isolation & purification; Fundamental and applied biological sciences. Psychology; Liquid chromatography; Mobile phase additives; Optimization criteria; Other biological molecules; Temperature; Terpenes, steroids. Hormones; Mobile phase additives; Cyclodextrin; Temperature; Liquid chromatography; Optimization criteria; Estrogens; Capacity factor; Steroid; Oligosaccharide; Estrogen; HPLC chromatography; Environmental factor; Mobile phase; Retention time; Optimization; Separation method; Reversed phase chromatography; Concentration effect; Sex steroid hormone; Inclusion compound; Quantitative analysis
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/S0731-7085(96)01974-7

The effect of temperature on the retention and multiple separation of six estrogenic steroids in reversed-phase liquid chromatography has been studied. Capacity factors (k′) of estriol, 17β-estradiol, 17α-estradiol, d-equilenin, equilin and estrone were measured using mobile phase modified with different concentrations of β-cyclodextrin (from 0–16 mM), a fixed solvent composition (acetonitrile-water) and a wide range of column temperatures (from 5 to 80°C). The plots of capacity factors vs. reciprocal of absolute temperature are nonlinear in each case when mobile phase modified with β-cyclodextrin was used. Particularly strong nonlinearity was observed at lower temperature and at higher β-cyclodextrin concentration. The complex chromatograms were evaluated using optimization parameters such as capacity factor of the last-eluted peak (
k′
max), the smallest resolution between adjacent peaks (
R
s,min) and relative resolution product (
r). The results presented describe precisely the role of temperature in high-performance liquid chromatography systems in which mobile phases modified with cyclodextrin were used. Moreover, the elution order of estrogenic steroids on modified and unmodified mobile phases has been discussed.