title: The influence of temperature on the high performance liquid chromatographic separation of steroids using mobile phases modified with beta-cyclodextrin.
abstract: The effect of temperature on the retention and multiple separation of hydrocortisone, testosterone, 17 alpha-methyltestosterone, prednisone, cortisone and 17 alpha-hydroxyprogesterone in reversed-phase liquid chromatography has been studied. Capacity factors (k') of the steroids were measured using a mobile phase modified with different concentrations of beta-cyclodextrin (from 0-16 mM), a fixed solvent composition (acetonitrile-water) and a wide range of column temperatures (from 5-80 degrees C). The plots of capacity factors vs. reciprocal of absolute temperature are nonlinear in every case when mobile phase modified with beta-cyclodextrin was used. Particularly strong nonlinearity was observed at lower temperature and at higher beta-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 (Rs; 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.
authors.affiliation: Medical University of Gdańsk, Faculty of Pharmacy, Poland.
authors.forename: P K
authors.initials: PK
authors.lastname: Zarzycki
authors.forename: M
authors.initials: M
authors.lastname: Wierzbowska
authors.forename: H
authors.initials: H
authors.lastname: Lamparczyk
mesh_terms: D019326:17-alpha-Hydroxyprogesterone / Q000032:analysis / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003348:Cortisone / Q000032:analysis / Q000302:isolation & purification; D003505:Cyclodextrins; D006854:Hydrocortisone / Q000032:analysis / Q000302:isolation & purification; D007202:Indicators and Reagents; D007700:Kinetics; D008777:Methyltestosterone / Q000032:analysis / Q000302:isolation & purification; D011241:Prednisone / Q000032:analysis / Q000302:isolation & purification; D011283:Pregnenes / Q000032:analysis / Q000302:isolation & purification*; D013696:Temperature; D013739:Testosterone / Q000032:analysis / Q000302:isolation & purification*; D047392:beta-Cyclodextrins
chemical_list: D003505:Cyclodextrins; D007202:Indicators and Reagents; D011283:Pregnenes; D047392:beta-Cyclodextrins; D013739:Testosterone; D019326:17-alpha-Hydroxyprogesterone; C031215:betadex; D003348:Cortisone; D008777:Methyltestosterone; D011241:Prednisone; D006854:Hydrocortisone