Abstract
The unusual temperature effect on the retention of 17α-estradiol, 17β-estradiol and equilin in reversed-phase liquid chromatography has been observed. Capacity factors ( k′) of the steroids were measured using mobile phase with different concentrations of β-cyclodextrin (from 0 to 16 mM) in a fixed mobile phase composition (acetonitrile-water) and 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. At subambient temperatures the capacity factors decreased with temperature decrease. This effect is more evident for the natural 17β-estradiol than for its 17α-isomer.
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