HPLC of Steroids in Non-aqueous Mobile Phase at Subambient Temperature

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Abstract
Increased resolution of steroid mixtures was found in high performance liquid chromatography of steroids at subambient temperatures. With aqueous mobile phase using a reversed phase column it was not possible to decrease temperatures below −10°C due to increased viscosity. This report describes further increase in resolution at −50°C using a non-aqueous mobile phase for the separation of steroids. Retention times were shorter several fold while resolution was improved.