A MODIFIED METHOD FOR THE ANALYSIS OF URINARY 17-KETOSTEROIDS, PREGNANEDIOL AND PREGNANETRIOL BY GAS LIQUID CHROMATOGRAPHY IN NORMAL SUBJECTS AND SUBJECTS WITH VARIOUS ENDOCRINE DISORDERS

Abstract

ABSTRACT A modified method using gas-liquid chromatography (GLC) is described for the determination of androsterone*, aetiocholanolone, dehydroepiandrosterone, pregnanediol and pregnanetriol in the same urine aliquot. After sequential enzymatic hydrolysis and solvolysis, the urine extract is separated into 3 main fractions by silica gel column chromatography; one containing mainly the 17-ketosteroids, androsterone, aetiocholanolone and dehydroepiandrosterone; a second containing pregnanediol and the third pregnanetriol. After formation of the trimethylsilyl ethers (TMSi), the respective fractions are subjected to gas-liquid chromatography for quantitation. Tritiated androsterone, pregnanediol and pregnanetriol are added prior to extraction of the urines to check for recoveries. The method is specific and reproductible and is sufficiently sensitive to permit determination of urinary steroid excretion levels as low as 0.025 mg/24 hours. Results obtained with this method in the study of normal males and females and various pathological conditions are presented.