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Clinical Chemistry 10: 170-183, 1964;
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Clinical Chemistry, Vol 10, 170-183, Copyright © 1964 by the American Association for Clinical Chemistry

Urinary Pregnanediol and Pregnanetriol: Spectrophotometric Determination Following Aluminum Oxide Column Chromatography

Daniel Klein 1 and Joseph M. Chernaik 1

1 Long Island Laboratories, Inc., Flushing 54, N. Y.

A modification of the procedure for urinary pregnanediol and pregnanetriol is described employing enzymatic hydrolysis, solvent extraction, separation on aluminum oxide by partition gradient elution, and determination by use of the sulfuric acid color reaction. A procedure has been devised for overcoming chromogenic interference by impurities, especially with the pregnanediol determination, based on utilization of the absorption curve characteristics of these two steroidal chromogens in H2S04. One essential step is the incorporation of the standards into one of a pair of urine extracts; a second is the collection of a small number of eluates for both the pregnanediol and pregnanetriol fractions. By the wavelength of peak absorption obtained in H2S04 for each eluate residue, the proper eluates are then selected for the determination of each of the two steroids. The method requires simple equipment readily available in clinical chemistry laboratories, and gives results that compare very favorably with those obtained with other procedures.

Submitted on March 21, 1963






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Copyright © 1964 by the American Association for Clinical Chemistry.