Studies on the High Voltage Paperelectrophoresis of Adrenocortical Steroids and their Related Compounds<subtitle>II. Some Conditions for the Electrophoretic Separation of Cortisone, Hydrocortisone and their 21-Acetates</subtitle>

Abstract

HVPE separtion of the authentic samples of cortisone, hydrocortisone and their acetates were attempted. The electrophoretic conditions were studied in respect to the composition of the buffer solution. The results are summarized as follows. Cortisone acetate and hydrocortisone acetate did not migrate at all in any buffer medium used in the present study. Cortisone and hydrocortisone migrated at reasonable velocities in neutral (pH 7.0) and basic buffer (pH 10.0) hut not in acidic buffer (pH 1.5). The use of neutral buffer gave the highest mobility as epxress ed by RLys values. The addition of CMC to the electrophoretic buffer accelerated the mobility and RLysvalues of cortisone and hydrocortisone. The effect was greatest when CMC was added to neutral medium. The mobility and RLys values of hydrocortisone was higher than those of cortisone in any kind of buffer. But when hydrocortisone and cortisone were mixed together, these two steroids were not well separated but it gave a single band with a RLys value close to that of hydrocortisone. Cortisone acetate and hydrocortisone acetate remained at original line even in admixture with the respective free forms which migrated in the same manner as in their individual runs. Test for the mutual interference using 2 free and 2 conjugated steroids on the HVPE patterns revealed that it occurred only between cortisone and hydrocortisone.