Determination of Testosterone in Human Peripheral Blood Using Gas-Liquid Chromatography with Electron Capture Detection

Brownie, Alexander C.; Molen, H. J. Van Der; Nishizawa, Edward E.; Eik‐Nes, Kristen B.

doi:10.1210/jcem-24-11-1091

Cited by 123 publications

(28 citation statements)

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“…Measurements of testosterone were done with the method of Brownie, van der Molen, Nishizawa and Eik-Nes (22). Androstenedione was reduced to testosterone and then determined by the same method (23).…”

Section: Plasma Levelsmentioning

confidence: 99%

Androgen Production and Conversion to Estrogens in Normal Postmenopausal Women and in Selected Breast Cancer Patients

Poortman

Thijssen

Schwärz

1973

The Journal of Clinical Endocrinology & Metabolism

106

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Urinary production rates of dehydroepiandrosterone (D) and dehydroepiandrosterone sulfate (DS) by isotope dilution, and the blood production rate of androstenedione (A) by continuous infusion technique have each been measured in 8 normal postmenopausal women. Both measurements were done in a group of 6 breast cancer patients. The cancer patients were at the time of the primary operation at least 6 yr postmenopausal; at the time of the study they were at least 3 yr after the operation. All of them were in good health and free of demonstrable recurrence of the tumor.The contribution of D and DS to urinary estrogens and the conversion of A to estrone were each determined in 8 normal postmenopausal women. Both measurements were done in the 6 breast cancer patients.The mean urinary production rate of D and DS were lower in the breast cancer patients than in the normal subjects. The difference was significant (p < 0.025) for DS. In the breast cancer patients the excretion of ll-deoxo-17-oxosteroids (11-DOKS) was lower than in the normal subjects. It is highly probable that the decreased production of D and DS is the cause of the low excretion of 11-DOKS in the patients.In 6 of the 14 experiments the specific activity of D isolated from the sulfate and the glucuronide fraction was not consistent with the model of van de Wiele et al. It is likely that in these cases neither D-glucuronide nor D-sulfate could be considered a unique specific metabolite of the D-and the DS-pool respectively. The contribution of D and DS to urinary estrogens was low (less than 0.01%) in both normals and in breast cancer patients.In the normal subjects we found a mean metabolic clearance rate of A of 1843 ± 131 (SE) liter per day and a mean plasma-level of A of 0.83 ± 0.13 ng per liter leading to a daily blood production rate of 1.64 ± 0.38 mg of A. In the breast cancer patients we found a mean metabolic clearance rate of 2398 ± 4 2 5 (SE) liter per day and a mean plasma-level of A of 0.60 ± 0.12 \xg per liter leading to a daily blood production rate of 1.46 ± 0.39 (SE) mg of A.The conversion of circulating androstenedione to estrone was 2.5 ± 0.3 ( S E ) % in the normal subjects and 2.9 ± 0.2 ( S E ) % in the breast cancer patients. Both in the normal subjects and in the breast cancer patients a daily estrone production of approximately 40 M*g can therefore be accounted for by the conversion of plasma born androstenedione. (/ Clin Endocrinol Metab 37: 101, 1973)

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Section: Plasma Levelsmentioning

confidence: 99%

Androgen Production and Conversion to Estrogens in Normal Postmenopausal Women and in Selected Breast Cancer Patients

Poortman

Thijssen

Schwärz

1973

The Journal of Clinical Endocrinology & Metabolism

106

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“…The lowthroughput, gas-liquid chromatographic procedures formerly used for the assay of plasma testosterone (Brownie et al, 1964;Sarda and Strauss, 1968) have now been replaced by sensitive radioimmunoassay (RIA) techniques. Specific antisera eliminate the need for time-consuming pre-assay purification procedures (Hillier et al, 1973a;Falvo and Nalbandov, 1974), but the use of tritiated radioligands and liquid scintillation cocktail add substantially to the running costs of the assay.…”

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confidence: 99%

A Simple Robust Assay for Testosterone in Male Plasma Using an¹²⁵I-Radioligand and a Solid-Phase Separation Technique

1979

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A radioimmunoassay for testosterone in male plasma utilising a gamma-emitting radioligand and a solid-phase antiserum is described. The radioligand is testosterone-3-(O-carboxymethyl)-oxime coupled to 125I-iodohistamine, and the solid-phase antiserum is prepared by coupling antitestosterone-3-bovine serum albumin to cyanogen bromide activated cellulose. The new procedure retains much of the specificity associated with a published, specific radioimmunoassay using an antiserum raised against testosterone-11 alpha-BSA and a tritium radioligand and incorporating a dextra-coated charcoal separation procedure; values obtained by the two procedures are in excellent agreement (r = 0.98, n = 20). The combination of an 125I-radioligand and a solid-phase separation technique greatly increases sample throughput and has the further advantage of reduced running costs and a greater potential for automation. The method gives satisfactory levels of sensitivity, precision, and accuracy.

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“…The samples were then centrifuged, and the plasma stored at -15°C until assayed. Plasma levels of testosterone and progesterone were determined by gas-liquid chromatog raphy with electron capture, using the methods of Brownie et al [1965] and Van der Molen and G roen [1965], respectively. Plasma corticosterone was determined by a modifi cation of the protein-binding method of M urphy [1967], while serum LH was measured by the radioimmunoassay method of Scaramuzzi et al [1972].…”

mentioning

confidence: 99%

Effects of an Intraventricular Injection of Synthetic ACTH on Plasma Testosterone, Progesterone and LH Levels and on Sexual Behavior in Male and Female Rabbits

Haun¹,

Haltmeyer²

1975

Neuroendocrinology

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Sexual behavior and elevation of gonadal steroids were induced in both male and female New Zealand white rabbits following injection of synthetic β^(1–24) ACTH into the lateral cerebral ventricle. In blood assays collected by cardiac puncture, testosterone, progesterone and corticosterone were all increased above both resting levels and samples were taken after a control intraventricular injection of physiological saline. Parallel increases of plasma LH were also demonstrated. The results were interpreted to indicate that intraventricular ACTH, through LH release, influences sex hormone levels and sexual behavior by acting ‘centrally’, probably within the hypothalamus.

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Determination of Testosterone in Human Peripheral Blood Using Gas-Liquid Chromatography with Electron Capture Detection

Cited by 123 publications

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Androgen Production and Conversion to Estrogens in Normal Postmenopausal Women and in Selected Breast Cancer Patients

Androgen Production and Conversion to Estrogens in Normal Postmenopausal Women and in Selected Breast Cancer Patients

A Simple Robust Assay for Testosterone in Male Plasma Using an¹²⁵I-Radioligand and a Solid-Phase Separation Technique

Effects of an Intraventricular Injection of Synthetic ACTH on Plasma Testosterone, Progesterone and LH Levels and on Sexual Behavior in Male and Female Rabbits

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Determination of Testosterone in Human Peripheral Blood Using Gas-Liquid Chromatography with Electron Capture Detection

Cited by 123 publications

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Androgen Production and Conversion to Estrogens in Normal Postmenopausal Women and in Selected Breast Cancer Patients

Androgen Production and Conversion to Estrogens in Normal Postmenopausal Women and in Selected Breast Cancer Patients

A Simple Robust Assay for Testosterone in Male Plasma Using an125I-Radioligand and a Solid-Phase Separation Technique

Effects of an Intraventricular Injection of Synthetic ACTH on Plasma Testosterone, Progesterone and LH Levels and on Sexual Behavior in Male and Female Rabbits

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A Simple Robust Assay for Testosterone in Male Plasma Using an¹²⁵I-Radioligand and a Solid-Phase Separation Technique