[A simple method for the estimation of a urinary steroids profile using glass capillary gas chromatography with a...

A simple method for the estimation of a urinary steroids profile with glass capillary gas chromatography is described. A 10ml portion of a urinary specimen was hydrolysed by Helix pomatia juice. Additional hydrolysis with ox-liver beta glucuronidase was necessary for the complete liberation of the urinary steroids from their glucuronides conjugates. Liberated free steroids were extracted with ethyl acetate, and the extract was washed and evaporated under reduced pressure. Finally extracted steroids were derivatised as MO-TMS (methoxime-trimethylsilyl) ethers. For the full-silylation of the steroids, the Sakauchi-Hornig method was employed and good results were obtained. An OV-101-WCOT column (30m length) was connected to the "solventless injection system" (a type of glass falling needle injection) and temperature-programmed gas chromatography was performed. By this method, eight fractions of 17-KS, Pregnanediol, Pregnanetriol, Pregnanetriolone, 5-Pregnentriol, tetrahydro-metabolites of 11-Deoxy-Cortisol and of Cortisol, and hexahydrometabolites of Cortisol were separated and quantitatively determined. Data processing was performed by an on-line microcomputer. The value in normal male and female controls was compatible with the results of glass capillary gas chromatography measured by Shackleton. Urine specimens from children with adrenocortical cancer and 21-hydroxylase deficiency were analyzed. Characteristic metabolic profiles of each patient were easily demonstrated. This method seems suitable for the routine clinical elucidation of abnormal steroid metabolism.