Steroids, Urinary

Determination of urinary steroids has been of continued interest because (a) certain 

The gas-phase analytical techniques have been used for the analysis of urinary steroids for a long time. The determinations of urinary estrogens, progesterone metabolites, 17-ketosteroids and, to a lesser degree, corticosteroid metabolites, with packed-column GC are extensively documented in the earlier monographs on the subject [274,290]. Various sample treatments, approaches to conjugate hydrolysis, and volatile derivatives have been described. Among those steroids, aldosterone stands out as a uniquely difficult substance to derivatize and determine. 

Whereas the earlier work used GC for the estimation of only selected urinary steroid constituents, most current work favors the multicomponent (metabolic profiling) approach. The preparation of mixed derivatives , such as, for example, the methoxime-trimethylsilyl derivatives of Gardiner and Homing [291], in principle facilitates conversion of all metabolites containing hydroxy and carbonyl groups. Through the introduction of capillary columns to steroid analysis [12,292,293], it became possible to separate the complex mixtures of such derivatives. 

Although the present use of urinary steroid profiling techniques appears to be largely confined to biomedical research, their gradual acceptance in clinical diagnosis and preventive medicine (to establish the risk of biochemical endocrine disorders) 

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Juricskay S, Telegdy E. 2000. Urinary steroids in women with androgenic alopecia. Clin Biochem 33 97. 

Drug/Lab test interactions Plasma corticosteroid levels may be increased. Urinary steroid determinations may be altered by amphetamines. 

B2. Barnard, G., and Kohen, E., Monitoring ovarian function by the simultaneous time-resolved fluorescence immunoassay of two urinary steroid metabolites. Clin. Chem. 44,1520-1528 (1998). 

After metyrapone administration, a patient with a disease of pituitary origin cannot achieve a compensatory increase in the urinary excretion of 17-hydroxycorti-costeroids or 11-deoxysteroids. Moreover, if pituitary corticotrophin is suppressed by an autonomously secreting adrenal carcinoma, there will be no increase in response to metyrapone. On the other hand, if pituitary corticotrophin secretion is maintained, as occurs in adrenal hyperplasia, the inhibition of corticoid synthesis produced by metyrapone will stimulate corticotrophin secretion and the release of metabohtes of precursor urinary steroids, which can be measured as 17-hydroxycortico-steroids. Metyrapone is now used less frequently in the differential diagnosis of Cushing s syndrome because of the ability to measure plasma corticotrophin directly. 

Patients with a complete block have almost undetectable excretions of cortisol metabolites. The major urinary steroids are androsterone, etiocholanolone, tetrahydro-... 

Table 5.3.6 Diagnostic urinary steroid ratios for 5a-reductase deficiency (homozygous and heterozygous), and a patient with 5 -reductase deficiency.

We found normal levels of conventional urinary steroids in this disorder, but these patients can show blunted response to ACTH, and adrenal hyperplasia is often noted post-mortem in nonsurviving newborns. Many distinctive metabolites have been found in urine from patients with the disorder, major examples being 5/3-prcgn-... 

Glass IA, Lam RC, Chang T, Roitman E, Shapiro LJ, Shackleton CH (1998) Steroid sulphatase deficiency is the major cause of extremely low oestriol production at mid-pregnancy a urinary steroid assay for the discrimination of steroid sulphatase deficiency from other causes. Prenat Diagn 18 789-800... 

Honour JW, Tourniaire J, Biglieri EG, Shackleton CHL (1978) Urinary steroid excretion in 17a-hydroxylase deficiency. J Steroid Biochem 9 495-505... 

Shackleton CH (1976) Congenital adrenal hyperplasia caused by defect in steroid 21-hydroxylase. Establishment of definitive urinary steroid excretion pattern during first weeks of life. Clin Chim Acta 67 287-298... 

Shackleton CH (1986) Profiling steroid hormones and urinary steroids. J Chromatogr 379 91-... 

Shackleton CH, Taylor NF, Honour JW (1980b) An Atlas of Gas Chromatographic Profiles of Neutral Urinary Steroids in Health in Disease. Packard-Becker, Delft... 

Shackleton CH, Roitman E, Kelley R (1999) Neonatal urinary steroids in Smith-Lemli-Opitz syndrome associated with 7-dehydrocholesterol reductase deficiency. Steroids 64 481-490... 

Although it has been stated that only phenols are conjugated with sulfuric acid,1 three urinary steroid sulfate esters (androsterone sulfate, dehydro-epi-androsterone sulfate, and A 6-aZZo-pregnene-3(/3)-ol-20-one sulfate) have been isolated in which the sulfuric acid is esterified by an alcoholic hydroxyl group.111 It is true that no alcohol administered to... 

Moon J, Ha Y, Moon M, Chung B, Choi M (2010) Systematic error in gas chromatography-mass spectrometry based quantitation of hydrolyzed urinary steroids. Cancer Epidemiol Biomarkers Prev 19 388-397... 

Phenothiazines can interfere with the measurement of urinary steroids (colorimetry) or oxosteroids (Porter-Silber test) (SED-11,115). 

Figure 1.7. Reparation of trimethylsilyl ethers of infant urinary steroids on 1) packed (9 ft X 4 mm, 2% SE-30, 190-275°C programmed at l.5"C/min) and 2) open tubular (80ft X 0.3 mm, OV-101, 160-270 C programmed at 2.5°Clmin for 30 min and l°C/minfor...

Detection and quantitation of steroids. The confirmation of the steroid content of a biological sample is a routine procedure in many GC-MS laboratories. The development of the human urinary steroid profiles ... 

The most striking aspect of the endocrine response involves the adrenal cortex. Albright was among the first to observe a raised output of urinary steroids in injured patients and to note the similarity between the postoperative response and the changes of Cushing s syndrome (A2). 

The usefulness of plasma cortisol measurements and urinary steroid levels is restricted because of the rapid fluctuations in hormone levels in blood and the possible important variations within different groups of urinary steroids which cannot be detected when only total excretion is measured. 

B26. Bush, I. E., Automation of the analysis of urinary steroids using quantitative paper chromatography and a small laboratory digital computer. Clin. Chem. 14, 491-512 (1968). 

Wudy SA, Homoki J, Wachter UA, Teller WM. Diagnosis of the adrenogenital syndrome caused by llp-hydroxylase deficiency using gas chromatographic-mass spectrometric analysis of the urinary steroid profile. Dtsch Med Wochenschr 1997 122 3-11. 

Aguilera R, Becchi M, Casablanca H> Hatton CK, Gatlin DH, Starcevic B, et al. Improved method of detection of testosterone abuse by gas chromatography/combustion/isotope ratio mass spectrometry analysis of urinary steroids. J Mass Spectrom 1996 31 169-76. 

Examples of possible uses of biological materials (pooled collections of scrum, urine, etc.) as standards are in methods for estimating total protein concentration in serum, for measuring enzyme activities in serum and other body fluids, and for determining urinary steroids and their metabolites. The special requirements of multichannel analysis will be considered separately (Section 3.1.1.3), and the reasons why these three different investigations can perhaps best be standardized in relation to biological materials will now be discussed. 

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. 

After the development of the method for measuring urinary steroids, Daughaday discovered elevated steroids in the urine of patients with Cushing s disease. Finally, the end product was identified and Cushing s syndrome was correctly explained as an excess of cortisol in the plasma (hypercortisolism). 

Mitotane appears to be the drug of choice in inoperable functional and nonfunctional adrenal carcinoma. Tumor regression is seen in approximately 35% to 50% of patients, with most regression occurring between the second and fourth month of therapy. Seventy-five percent of patients will exhibit a 30% fall in urinary steroids, with 50% of patients showing an improved clinical response after 5 months of... 

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