PAPIERELEKTROPHORESE VON STEROIDEN

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Abstract
Steroids are esterified with a slight excess of succinic anhydride in pyridine for 18 hrs. at 60[degree] C in a reflux flask. The pyridine is removed in vacuo, the residue dissolved in 90% C2H5OH, and brought to pH of 7.05-7.15 with alcoholic NaOH. After standing 18 hrs. at room temp. the specimen is dried in vacuo. Blank controls are run with pure succinic anhydride. The residue is taken up in 0.5% concn. in a pH 7.5 phosphate buffer (ionic strength .066), 80% and 20% C2H5OH. Paper strips 36 x 4 cm. are moistened in the buffer, blotted and the soln. of steroid ester (0.1 mg./.02 ml.) placed along a line at one end. Electrophoresis is carried out with previously descr. apparatus. The strips are dried (100[degree]C) and developed with 15% H3PO4, m-dinitrobenzene or triphenyltetrazolium chloride. The method successfully separates desoxycorticosterone from dehydroepiandrosterone.