Factors affecting the excretion of urinary steroid metabolites in man : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Biochemistry at Massey University

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Massey University
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A gas liquid chromatograph (GLC), adapted to accept a "Support Coated Open Tubular" (SCOT) capillary column, was used for profiling of the neutral steroid hormone metabolites from human urine specimens. Following hydrolysis with ß-glucuronidase to release the steroid metabolites from their conjugates, they were extracted with organic solvents and converted into methoxime-trimethylsilyl derivatives for separation on the GLC column. Initial work involved determining basal data on consecutive days and also during the twenty four hour period. The circadian variation in this experimental work, showing a biphasic pattern of steroid metabolite excretion, with a maxima in the morning, and a second smaller maxima in the afternoon. The effect of cold stress on metabolite excretion was examined, however the results obtained were inconclusive. Dexamethasone was administered, 0.5 mg per six hours for forty eight hours, to induce suppression of ACTH secretion. The expected pattern of decreased 17 hydroxycorticoid excretion, and to a lesser extent 17 oxosteroid excretion was observed. Alcohol loading was examined, with varying levels of intoxication. One subject maintained a blood alcohol level of approximately 50 mg% for three hours, and the urine specimens were collected at two hour intervals. The excretion rate of the steroid metabolites increased a few hours after ethanol loading, compared to the control. THE, THF and aTHF showed the greatest increments, and this may possibly be due to increased hepatic A ring reductase activity resulting from increased NADH:NAD+ ratio. A detailed examination of excretion rates and plasma cortisol concentration would be required to prove that ethanol causes this action on the A ring reductase activity. Three twenty-four hour specimens were collected from a subject with a history of alcohol abuse over the previous ten years. On Day 2 of the three day experiment, a 26 ounce bottle of Scotch was consumed, but other than an increase ot all steroid metabolites on the day of alcohol loading, no significant observations were made. Three urine specimens from alcoholics admitted to the Palmerston North Detoxification Unit were analysed. No llOHEt could be detected in any of these three patient's urine, however the ß-glucuronidase used in this analysis had a low activity resulting in incomplete hydrolysis, and basal data for these patients would be required to prove that the absence of llOHEt was a result of alcohol intoxication. One of the alcoholic patients showed high levels of cortisol metabolites, probably the result of stress of hospitalization.
Steroid hormones, Metabolism