The influence of temperature on the high performance liquid chromatographic separation of steroids using mobile phases modified with beta-cyclodextrin.
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.
Zarzycki
P K
PK
Medical University of Gdańsk, Faculty of Pharmacy, Poland.
Wierzbowska
M
M
Lamparczyk
H
H
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
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