Methods of Extracting Compounds Related to the Steroid Hormones From Human Urine

Callow, N. H.; Callow, R. K.; Emmens, C. W.; Stroud, Sally

doi:10.1677/joe.0.0010076

Cited by 70 publications

(16 citation statements)

References 5 publications

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“…Each individual fraction from the chromatogram was analyzed by infrared spectrometry and quantitation of the separated steroids was done by the Zimmerman reaction (as modified by Callow, Callow, Emmens, and Stroud [3,4] and Talbot, Butler, MacLachlan, and Jones [5]). Since ketosteroids differ somewhat in chromogenic value, appropriately determined factors were used to correct the color values from milligrams of dehydroisoandrosterone, used as the standard, to milligrams of the actual compound determined in the chromatographic fraction.…”

Section: Methodsmentioning

confidence: 99%

STUDIES IN STEROID METABOLISM. XIX. THE a-KETOSTEROID EXCRETION PATTERN IN NORMAL MALES 1

Dobriner¹,

Kappas²,

Rhoads³

et al. 1953

J. Clin. Invest.

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The urinary steroids afford a means for evaluation of the production of steroid hormones by the adrenal and gonads. Any alteration in either rate of type of hormone production by the glands resulting from specific disease or from hormonal therapy therefore must be manifest by qualitative or quantitative change in the excretion of steroid hormone metabolites. In order to delineate the composition of these metabolites in normal healthy subjects, the present investigation describing the a-ketosteroid excretion pattern was undertaken. MATERIALS AND METHODSTwenty normal males, ranging in age from 21 to 76, were studied. Of this number, eighteen were White and one was Negro; Subject D31 was a Nisei. Subjects C21 and M21 were normal males hospitalized at the Peter Bent Brigham Hospital, Boston, Massachusetts, for the purpose of studying some of the metabolic effects of oral Cortisone and intravenous ACTH; Subjects K31, R32, R63, C64, M65, and M72 were subjects of similar study at the Baltimore City Hospitals, Baltimore, Maryland. Only control periods of these eight subjects are reported here. The remainder of the subjects are, or were, employees of this Institute, working and in apparent good health. The details of urine collection and handling in this laboratory have been reported previously (1).

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Section: Methodsmentioning

confidence: 99%

STUDIES IN STEROID METABOLISM. XIX. THE a-KETOSTEROID EXCRETION PATTERN IN NORMAL MALES 1

Dobriner¹,

Kappas²,

Rhoads³

et al. 1953

J. Clin. Invest.

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“…original urine were added to insure excess acid (22). The whole was boiled under a reflux condenser for half an hour, a time insuring maximum recovery of both estrogen and androgen (23,24,25 and androgen were placed in the ice box overnight to allow the water to drain from the walls of the separatory funnel, and the water was then removed. The androgenic ether fraction was evaporated to dryness under negative pressure on the water bath and the residue redissolved in 10 cc.…”

Section: Methodsmentioning

confidence: 99%

A Quantitative Study of the Urinary Excretion of Hypophyseal Gonadotropin, Estrogen, and Androgen of Normal Women

Werner¹

1941

J. Clin. Invest.

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and Surgeons, Work upon the assay of hormones in blood and urine has been progressive in volume and quality since Loewe (1) and Frank (2) demonstrated the presence of estrogenic hormone in blood. Loewe (3) then revealed that this substance is excreted cyclically in the urine. Such assays were made practicable by the work of Kahnt and Doisy (4) who standardized the procedure of assay for this hormone, using the changes induced in the vaginal smear of castrated adult rats. The presence of gonadotropic substance in blood and urine was established shortly thereafter. The existence of such a gonadotropin and its production in the anterior hypophysis were made clear by the fundamental work of P. E. Smith (5) and Zondek and Aschheim (6). They also provided a method for determining its presence by finding that premature maturity is induced in the immature rat and mouse by this hormone (6, 7). Aschheim and Zondek (8, 9), using this test object, were able to show that a gonadotropin may be detected in the urine of pregnancy, although this gonadotropin is now known to be chorionic in origin. Fluhmann (10), Zondek (11), and others soon pointed out that the hypophyseal gonadotropin is similarly excreted after ovariectomy and the menopause. The same gonadotropin next was found to appear during the middle of the normal menstrual cycle (12). The demonstration of the excretion of androgenic substances soon followed (13,14), and methods of assay were provided by the capon comb method (15) and the colorimetric reaction of Zimmermann (16). Finally, excretion products of progesterone were shown to appear in the urine (17,18 improvements in methods of extraction and assay now aid in such a study. The tannic acid precipitation procedure of Levin and Tyndale (19), and their method of assay, using the immature mouse uterine weight (20), make possible the quantitative assay of hypophyseal gonadotropin in normal urine. In addition, it has been found that estrogen and androgen may be quantitatively recovered from the supernatant and the alcohol-acetoneether washings of the tannic acid precipitate, as was observed by Freed and Hechter (21) after tungstic acid precipitation. Thus it is possible to assay the gonadotropin, estrogen and androgen in the same specimen of urine. Data are here presented from such a study of the complete urinary output of five normal women with regular menstrual cycles over a period of three to four months for each woman. An additional three cycles scattered throughout the year were examined in one of these cases. These assays have been directed towards determining the normal values and the pattern of hormone excretion in medically normal healthy women with regular cycles. This was thought to be essential for the later determination as to whether or not patients were excreting abnormal amounts of hormone. The interpretation of the results has suggested a possible mechanism affecting the hypophysis-ovary relation. METHODAll urine specimens were complete forty-eight-hour collections kept in tightly stoppered bottles ...

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“…we observe the fol lowing ratios from one publication. In thyrotoxicosis we have the colorimetric over biological units ratios of 104 (157). On the other hand another group reports the rather low values of 14 to 34 J.U.…”

Section: Separation Of Androgens and Related Compounds From Urinementioning

confidence: 95%

“…In the alternate procedure the properly acidified urine is boiled under reflux condenser with carbon tetrachloride for two hours. The treat ment with fresh carbon tetrachloride is repeated twice (151).…”

Section: Separation Of Androgens and Related Compounds From Urinementioning

confidence: 99%

Hormones

Koch¹

1940

Annu. Rev. Biochem.

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Because the more definitely biochemical advances in the field of hormones are very nearly limited to those of the steroid type together with the limited space available for reviewing a very extensive litera ture, I have taken advantage of the editors' liberal policy and have limited this review to the biochemistry of the steroid hormones. ESTROGENSDistribution and forms of estrogens.-Although all of the estro gen activity in cow and hog ovaries is considered to be in the free form, only 50 to 75 per cent is free in the horse ovary or its follicular fluid (1). During pregnancy 50 to 75 per cent of the blood estrogens in women are in the free form (2). The best method of hydrolysis of the conjugated forms of estrogens in pregnancy urine is still a prob lem. One g.roup recommends boiling the pregnancy urine for ten minutes with 15 per cent concentrated hydrochloric added. Longer heating results in serious loss of estrogens (3). Others recommend that the urine from pregnant mares be allowed to stand at pH 0.4 to 0.6 at room temperature for at least four weeks (4). The writer, in studies on urine from normal men and nonpregnant women, has obtained the highest yield of estrogens by boiling the urine previously acidified to pH 1 to 1.2 with hydrochloric acid for fifteen minutes.Longer boiling at this pH also causes destruction of estrogens. The gradual increase in the urinary excretion of estrogens in women to a peak of 80,000 rat units near the termination has been confirmed, but in one case of toxemia of pregnancy, the excretion was subnormal.

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Methods of Extracting Compounds Related to the Steroid Hormones From Human Urine

Cited by 70 publications

References 5 publications

STUDIES IN STEROID METABOLISM. XIX. THE a-KETOSTEROID EXCRETION PATTERN IN NORMAL MALES 1

STUDIES IN STEROID METABOLISM. XIX. THE a-KETOSTEROID EXCRETION PATTERN IN NORMAL MALES 1

A Quantitative Study of the Urinary Excretion of Hypophyseal Gonadotropin, Estrogen, and Androgen of Normal Women

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