A Simplified Method for the Simultaneous Determination of 17-Ketosteroids, Dehydroepiandrosterone, and 17-Hydroxycorticosteroids in Serum

Saier, Eleanor L.; Campbell, Elizabeth A.; Strickler, Herbert S.; Grauer, Robert C.

doi:10.1210/jcem-19-9-1162

Cited by 22 publications

(9 citation statements)

References 8 publications

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“…Although in a few cases Zimmermann chromo¬ gens were detected in that zone, the quantities were too small for an accurate estimate. These findings are in agreement with those of Baulieu (1960) and of Oertel & Eik-Nes (1961) and are in contrast to those of Oertel & Kaiser (1962) The normal values found fall within the range reported by most authors (Saier, et al 1959;Conrad et al 1961;Dray & Adam, 1962;Eberlein, 1963;Deshpande & Bulbrook, 1964). The figures obtained by Migeon & Plager (1955) and Migeon et al (1957) are distinctly lower, probably because of methodological differences.…”

Section: Discussionsupporting

confidence: 91%

“…Most methods for their determination involve hydrolysis of the conjugates. This was brought about by treatment with hot acid (Gardner, 1953), by continuous extraction of the acidified extract at room temperature (Migeon & Plager, 1955; Bongiovanni & Eberlein, 1957;Ceresa & Cravetto, 1958) and recently by solvolysis (Saier, Campbell, Strickler & Grauer, 1959;Hudson & Oertel, 1961;Dray & Adam, 1962) or transesterification (Eberlein, 1963). Direct determination of conjugated 17-oxosteroids (Conrad, Mahesh & Herrmann, 1961) does not seem to have been widely applied.…”

Section: Introductionmentioning

confidence: 99%

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The Determination of 17-Oxosteroid Sulphates in Human Plasma

Neve¹,

Vermeulen²

1965

Journal of Endocrinology

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An accurate, precise and specific method for the determination of plasma 17-oxosteroid sulphates has been developed. It involves extraction of the steroid sulphates, solvolysis, purification by solvent partition and thi n\x=req-\ layer chromatography and determination by a modified Zimmermann reaction.In both sexes plasma 17-oxosteroid levels were found to be low before puberty, to reach their highest level between the ages of 20 and 30 and to decrease rapidly after the age of 35. Maximal levels in females were only about 70% of those in males; after the age of 35 the plasma levels were similar in both sexes. The proportions of the individual 17-oxosteroids (dehydroepiandrosterone, androsterone and aetiocholanolone) were not influenced by sex; in ten obese patients of both sexes the percentage contribution of dehydroepiandrosterone was significantly decreased.

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Section: Discussionsupporting

confidence: 91%

Section: Introductionmentioning

confidence: 99%

The Determination of 17-Oxosteroid Sulphates in Human Plasma

Neve¹,

Vermeulen²

1965

Journal of Endocrinology

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“…The values for the concentration of the 17-oxosteroids in plasma found in this study are considerably higher than those reported by Migeon (1955), Migeon & Plager (1955) and Migeon et al (1957) but agree well with those found in other studies (Saier, Campbell, Strickler & Grauer, 1959;Conrad, Mahesh & Herrmann, 1961) in which dehydroepiandrosterone and androsterone sulphates were measured as the major components of the total 17-oxosteroids in plasma. It may be that the acid hydrolysis used by Migeon and his associates destroys some of the plasma steroids.…”

Section: Discussionsupporting

confidence: 90%

Simultaneous Determination of 17-Oxosteroids and 17-Hydroxycorticosteroids in Plasma From Normal Women

Deshpande¹,

Bulbrook²

1964

Journal of Endocrinology

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The method of Deshpande & Bulbrook (1964) for the determination of total 17-oxosteroids and 17-hydroxycorticosteroids (17-OHCS) in blood has been employed in a study of fifty-two normal women of different ages. The results show that there is a significant correlation between age and the plasma levels of 17-oxosteroids and 17-OHCS. Contrary to similar studies on urines, no other parameter gave a significant correlation. Factors which might control the relative amounts of plasma 17-oxosteroids and 17-OHCS are discussed.

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“…Recent experience of Finkelstein, Forchielli and Dorfman (1) suggests that in these cases the blood levels and nature of the circulating androgens may be very important. Earlier indirect evidence (2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12) and the more recent isolation and partial identification of sulfates of dehydroisoandrosterone, androsterone, and etiocholanolone by Baulieu (13) indicate that 17-ketosteroids are present in plasma mainly in the form of esters. Direct estimation of these conjugated steroids in plasma offers distinct advantages: the alteration of the steroid molecule which occurs with certain types of hydrolysis as well as possible problems of incomplete hydrolysis can be circumvented.…”

mentioning

confidence: 99%

Quantitative Paper Chromatography of Conjugated 17-Ketosteroids in Plasma*

Conrad¹,

Mahesh²,

Herrmann³

1961

J. Clin. Invest.

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The determination of urinary 17-ketosteroids has limited applications in the understanding and diagnosis of the nature of endocrine disorders associated with mild hirsutism and virilism in the female, because a very large number of these patients excretes 17-ketosteroids well within the normal range. Recent experience of Finkelstein, Forchielli and Dorfman (1) suggests that in these cases the blood levels and nature of the circulating androgens may be very important. Earlier indirect evidence (2-12) and the more recent isolation and partial identification of sulfates of dehydroisoandrosterone, androsterone, and etiocholanolone by Baulieu (13) indicate that 17-ketosteroids are present in plasma mainly in the form of esters. Direct estimation of these conjugated steroids in plasma offers distinct advantages: the alteration of the steroid molecule which occurs with certain types of hydrolysis as well as possible problems of incomplete hydrolysis can be circumvented. Also, information concerning the physiological significance of the steroid conjugating mechanisms may be gained through direct qualitative and quantitative estimation of these conjugates. The present paper describes a convenient method, using quantitative paper chromatography, for the estimation of dehydroisoandrosterone and androsterone sulfates (DHIA-SO and Andro-SO4) in plasma. MATERIALS AND METHODAll reagents and solvents used were of analytical grade, and all solvents, except absolute ethanol, were redistilled prior to use. Florisil was purified as described earlier (14). M-dinitrobenzene was purified by sublimation, using a modification of the "cold finger" technique, and stored in the dark at room temperature. Collection of plasm1ia. Blood from fasting subjects was drawn into heparinized syringes between 8:30 and 9: 30 a.m. The blood was then transferred to a glass centrifuge bottle containing 1 ml of heparin per 100 ml of blood and centrifuged at 2,500 rpm for 30 minutes. The plasma was removed with a syringe equipped with a long spinal needle and transferred to a clean glass vessel.Extraction of steroid con jngates. All analyses were run in triplicate. Fifteen-ml aliquots 1 of the plasma were placed in three 250-ml Erlenmeyer flasks and 45 ml of absolute ethanol was added to each flask. The flasks were swirled several times and placed in an ice bath for 10 to 15 minutes. After filtering the mixture, under vacuum, through two discs of Whatman no. 1 filter paper, the flasks were rinsed three times with 10 ml of absolute ethanol, and these washings were then used to wash the precipitate. The filtrate was transferred quantitatively to a 1 L round-bottomed flask with further washings of absolute ethanol and evaporated to near dryness (1 to 2 drops) on a flash evaporator (Laboratory Glass and Instruments Corp., model FE-2) under vacuum, using dry ice in ethylene glycol for refrigeration. Secondary butanol (15 ml) was then added to effect a partial separation of steroid conjugates from inorganic salts. The steroid conjugates, which dissolve quantita...

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A Simplified Method for the Simultaneous Determination of 17-Ketosteroids, Dehydroepiandrosterone, and 17-Hydroxycorticosteroids in Serum

Cited by 22 publications

References 8 publications

The Determination of 17-Oxosteroid Sulphates in Human Plasma

The Determination of 17-Oxosteroid Sulphates in Human Plasma

Simultaneous Determination of 17-Oxosteroids and 17-Hydroxycorticosteroids in Plasma From Normal Women

Quantitative Paper Chromatography of Conjugated 17-Ketosteroids in Plasma*

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