Dehydroepiandrosterone, in urine

Androsterone, Etiocholanolone, and Dehydroepiandrosterone in Urine Methoden Hormonbestimmung 1975 274-284 ... 

N, L., and Dorfman, R. I. Determination of Androsterone, Etio-cholanolone, and Dehydroepiandrosterone in Urine by Gas-Liquid Chr oma t o gr aphy... 

Rapid Gas Chromatographic Assay for Dehydroepiandrosterone in Urine Clin. Biochem. 4(4) 241-258 (1971) CA 76 123745V... 

According to the classical view of metabolism the hormones are synthesized as free steroids in endocrine tissues and prepared for excretion in urine by peripheral metabolism and conjugation. This view had to be modified upon the isolation of dehydroepiandrosterone sulfate from adrenal tumor [307]. Thus dehydroepiandrosterone sulfate, a steroid conjugate, was shown to be secreted by the adrenal tissue. Isotopic methods also pointed in the same direction. Lieberman et al. [304], using... 

After an injection of radioactive dehydroepiandrosterone sulfate, a small proportion is recovered as such in urine. However, if the double-labeled compound H-dehydroepiandrosterone sulfate- S is injected and its disappearance from the blood is observed, one finds different half-lives for H-dehydroepiandrosterone sulfate and for dehydroepiandrosterone sulfate- S (Fig. 10). This shows that there is a constant hydrolysis of the sulfo conjugate followed by a resulfation with sulfate from a large unlabeled pool (Fig. 3). In other words, the steroid moiety of dehydroepiandrosterone sulfate has a half-life of 9 hours, whereas the sulfate moiety has a shorter half-life of 6 hours, since the sulfate moiety is... 

Testosterone sulfate does not seem to be hydrolyzed in the organism, but its direct metabolites have not been identified, as only 3.5% of the radioactivity was found in urine after radioactive testosterone sulfate administration. Unlike dehydroepiandrosterone sulfate, testosterone sulfate is not transformed by indirect metabolism into estrogens during pregnancy (Dray, 1963). 

Dehydroepiandrosterone sulfate is the most important of the A -3/3-stenol sulfates present in urine. Its cleavage is accomplished much more readily than that of the saturated sulfates (Table III, methods B-E). However transformation products are readily formed and quantitative hydrolysis to dehydroepiandrosterone is not easily achieved. The transformation products derived from this sulfate will be discussed in detail in Section III, 4. 

Dehydroepiandrosterone is the principal 17-ketosteroid of the 3/3-hy-droxy series present in urine. The measurement of this fraction has been given certain diagnostic significance. For this reason it is particularly important that conditions of hydrolysis be chosen which give maximum conversion of dehydroepiandrosterone sulfate to dehydroepiandrosterone. If this is not the case, digitonin precipitation (Section VIII,2) will give erroneously low results and colorimetric assays could be in error as a result of differences in extinction coefficients of the transformation products present. 

Overall, other adrenal androgens also show a progressive decrease in urinary excretion in both men and women. Thus, the mean 17-ketosteroid urine levels of elderly people are about 50% of those in young adults. This is primarily secondary to decreased dehydroepiandrosterone (DHEA) and androsterone production. In men, about one-third of the daily 17-ketosteroids are of testicular origen, the remainder being mainly from the adrenals. Androstenedione is a prehormone for testosterone. 

Formation of sulfates [328] is relatively reversible. This may be due to the fact that the metabolic clearance rate [318] of sulfates is relatively low [389] and their renal excretion inefficient [4]. The conversion of dehydroepiandrosterone sulfate in vivo to androsterone and etiocholanolone glucuronide was demonstrated by Lieberman and co-workers, who administered isotopically labeled dehydroepiandrosterone sulfate and isolated labeled androsterone and etiocholanolone from glucuronicase-hydrolyzed urine [303]. Since in another study by Lieberman [174], the transconjugation from dehydroepiandrosterone sulfate to dehydroepiandrosterone glucuronide without free dehydroepiandrosterone intermediate was declared to be improbable, in vivo equilibrium between dehydroepiandrosterone sulfate and dehydroepiandrosterone seems probable [303]. 

In the third example, dehydroepiandrosterone is widely used for standardizing and for checking the performance of the Zimmermann reaction, which forms the final step in many methods of steroid estimation. There is, however, no suitable standard preparation including the commoner urinary steroids and their metabolites which can be carried through all the previous stages of analysis, and the only satisfactory form of standard material that includes the naturally occurring conjugates of the main steroids and their metabolites is urine collected from normal individuals or from patients treated with cortisol. 

Buiarelli et al. (2004) extended the above analytical approach to many more related steroids when they published a method for the direct analysis of 15 urinary anabolic steroids in a single run, namely T, epitestosterone, dehydroepiandrosterone (DHEA), androsterone, etiocholanolone, their sulfates and their glucuronides (Figure 2,2), They extracted 2 mL of human urine by solid-phase extraction with methanol elution and reconstituted the residue in aqueous methanol in the presence of deuterated internal standards (da-epitestosterone glucuronide, [16,16,17-"H3 testosterone sulfate and [16,16,17-2H3]testosterone), then monitored, for example, mJz. 289-97 and 109 for T and epitestosterone, miz 367-97 for their sulfates, and m/z 463-113 and 287 for their glucuronides. The method does not achieve quantitation, but it allows the estimation of ratios, which makes it possible to monitor the urinary steroid profile, which is useful for monitoring the abuse of anabolic steroids. 

Mixtures of the latter isomers which are not normally encountered in the urine of women, except in cases of ovarian tumors, could be studied. This was done by demonstrating uniquely dehydroepiandrosterone by the study of the spontaneous loss of water which is not observed in practice during the unimolecular decomposition of testosterone. 

It is possible to determine the presence of testosterone - 100 times lower than the dehydro compounds - in 2 pi of urine, i.e., 100 pg of free testosterone mixed with 100 ng of the dehydroepiandrosterone can be detected. These assays were made possible by the use of a calibration curve [210]. 

Androgens, the male sex hormones, proved far more elusive that either the estrogens and progestins since they occur at much lower concentrations in biological fluids. The bioassay used to track the isolation in this case comprised the capon unit . This was the amount of extract that produced a 20% increase in the surface of a rooster s comb. The 15 mg of pure crystalline testosterone isolated in 1931 came from about 15 0001 of urine. The structural investigations of this series relied on the then newly discovered side chain oxidations of cholestanol (13-1) (Scheme 1.13). This method in essence comprised fairly drastic oxidation of reduced cholesterols of known stereochemistry at the A-B junction to afford in fairly low yield products in which the side chain at Cn had been consumed to leave behind a carbonyl group. One of these products proved to be identical with androsterone (13-2). That compound had in turn been obtained from a sequence of reactions starting from dehydroepiandrosterone (13-3) that had been isolated from male urine. 

The earliest report of an isolated androgen was presented by Butenandt (10) in 1931. He isolated 15 mg of crystalline androsterone from 15,000 L of human male urine. A second crystalline compound, dehydroepiandrosterone, which has weak androgenic activity, was isolated by Butenandt and Dannenberg (11) in 1934. During the following year, testosterone was isolated from bull testes by David et al. (12). This hormone was shown to be 6 to 10 times as active as androsterone. 

The discovery of oestrone (VII) in 1929 by Doisy and independently by Butenandt was followed in 1934 by the isolation of progesterone (II) from corpus luteum tissue and of dehydroepiandrosterone (IV) from urine (see review by Petrow [16]). Although their chemical structures were not fully elucidated, a relationship between them and cholesterol was assumed. 

The bulk of dehydroxyepiandrosterone is converted to sulfate and excreted. However, the conjugation reaction is reversible, and consequently labeled androster-one is in the urine after injection of labeled dehydroxyepiandrosterone sulfate. Adrenal tissues can convert dehydroepiandrosterone to androstenedione or testosterone. 

The 17-ketosteroids increased in the urine include dehydroepiandrosterone, androsterone, etiocholano-lone, and 11-hydroxy- and 11-oxy-derivatives. Thus, it would appear that as a result of the block in 21-hydroxylation, the rate of conversion of 17-hydroxy-progesterone to carbon 19 compounds is increased. These results indicated that the adrenals of these patients are capable of 11-hydroxylations in spite of the fact that 21-hydroxylation does not take place. As a result of 11-hydroxylation, 11-ketosteroids, and especially 11-ketopregnaetriol, are excreted in excessive amounts in the urine of patients with adrenogenital syndrome. Thus, 17-hydroxyprogesterone is direct-... 

Roberts et ah (1964) answered part of the question by injecting doubly labeled cholesterol sulfate into the splenic artery (supplying 90% of the blood to an adrenal carcinoma) of a female patient and isolating various steroid sulfates in the urine of the first 24 hours. Of the recovered radioactivity 0.46% was dehydroepiandrosterone sulfate, bearing the same H/ S ratio as the injected compound. Urinary androstenediol-3-monosulfate and 5-pregnene-3/3,17a,20a-triol-3-monosulfate as well as 16a-hydroxydehydroepiandrosterone-3-monosulfate also had the same ratio as the injected cholesterol sulfate, demonstrating that all these compounds have a common sulfated precursor, which could possibly be cholesterol sulfate. 

This procedure has a specificity similar to that of the Munson reaction. It should be mentioned that, after the administration of certain steroids (133) and in certain diseases, substances responding to these tests appear in the urine. However, they clearly do not belong to the dehydroepiandrosterone complex. Therefore, a positive reaction with the reagents described above cannot be accepted as proof of the presence of dehydroepiandrosterone and of its known transformation products. 

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