Development and validation of a sensitive liquid chromatography–tandem mass spectrometry assay to simultaneously measure androgens and estrogens in serum without derivatization

Harwood, D. Tim; Handelsman, David J.

doi:10.1016/j.cca.2009.09.003

Cited by 224 publications

(159 citation statements)

References 43 publications

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“…While, similar to previous RCTs, we did not use clamp studies, we assessed glucose metabolism by two different methods that have reasonably good agreement with clamp studies (17,38,39). Importantly, in contrast to previous trials, we rigorously evaluated changes in body composition and assessed the effects of testosterone treatment on abdominal adipose tissue compartments.…”

Section: Discussionmentioning

confidence: 99%

“…Men aged 35-70 years of age were eligible to participate in this trial if they had a history of T2D, and the total testosterone (TT) level (averaged from two fasting morning specimens) was #12.0 nmol/L (346 ng/dL), as measured by electrochemiluminescence immunoassay (ECLIA). Although TT was measured by both ECLIA and liquid chromatography-tandem mass spectroscopy (LCMS/MS), recruitment was based on ECLIA because the LCMS/MS assay (17) was not available for routine clinical use. Therefore, samples were batched and measured by LCMS/MS at study end.…”

Section: Setting and Participantsmentioning

confidence: 99%

“…TT was measured by both ECLIA (3) and LCMS/MS (17), SHBG, and LH using ECLIA (3), and free testosterone was calculated according to Vermeulen's formula as described (3). Glucose, insulin, HbA 1c (3), and adiponectin (19) were measured as described.…”

Section: Schedule Of Assessments and Measurementsmentioning

confidence: 99%

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Effect of Testosterone Treatment on Glucose Metabolism in Men With Type 2 Diabetes: A Randomized Controlled Trial

et al. 2014

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OBJECTIVETo determine whether testosterone therapy improves glucose metabolism in men with type 2 diabetes (T2D) and lowered testosterone. RESEARCH DESIGN AND METHODSWe conducted a randomized, double-blind, parallel, placebo-controlled trial in 88 men with T2D, aged 35-70 years with an HbA 1c £8.5% (69 mmol/mol), and a total testosterone level, measured by immunoassay, of £12.0 nmol/L (346 ng/dL). Participants were randomly assigned to 40 weeks of intramuscular testosterone undecanoate (n = 45) or matching placebo (n = 43). All study subjects were included in the primary analysis. Seven men assigned to testosterone and six men receiving placebo did not complete the study. Main outcome measures were insulin resistance by homeostatic model assessment (HOMA-IR, primary outcome) and glycemic control by HbA 1c (secondary outcome). CONCLUSIONSTestosterone therapy does not improve glucose metabolism or visceral adiposity in obese men with moderately controlled T2D and modest reductions in circulating testosterone levels typical for men with T2D.Observational studies consistently show that 30-50% of men with type 2 diabetes (T2D) have lowered circulating testosterone levels, relative to references based on healthy young men (1-3). This association is independent of age and obesity (4,5), and low testosterone levels in men with T2D are independently associated with insulin resistance (3). However, it is not known whether low testosterone levels are a cause or a consequence of T2D or its associated clinical features. Several lines of evidence lend support to the hypothesis that testosterone treatment decreases insulin resistance. Experimental evidence, reviewed in Grossmann

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Section: Discussionmentioning

confidence: 99%

Section: Setting and Participantsmentioning

confidence: 99%

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Effect of Testosterone Treatment on Glucose Metabolism in Men With Type 2 Diabetes: A Randomized Controlled Trial

et al. 2014

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“…In postmenopausal women, the normal range for SHBG concentration is 21-149 nmol/l (RPAH, Australia). Testosterone, 17b-oestradiol (E 2 ), and DHT and its 3a-diol and 3b-diol metabolites were measured using LC-MS/MS, as described previously (28). The limit of detection for testosterone was 0.04 nmol/l, for E 2 3.4 pmol/l and for DHT 0.01 nmol/l.…”

Section: Methodsmentioning

confidence: 99%

Oral low-dose testosterone administration induces whole-body protein anabolism in postmenopausal women: a novel liver-targeted therapy

Birzniece

Umpleby

Poljak

et al. 2013

European Journal of Endocrinology

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Objective: In hypopituitary men, oral delivery of unesterified testosterone in doses that result in a solely hepatic androgen effect enhances protein anabolism during GH treatment. In this study, we aimed to determine whether liver-targeted androgen supplementation induces protein anabolism in GH-replete normal women. Design: Eight healthy postmenopausal women received 2-week treatment with oral testosterone at a dose of 40 mg/day (crystalline testosterone USP). This dose increases portal concentrations of testosterone, exerting androgenic effects on the liver without a spillover into the systemic circulation. Outcome measures: The outcome measures were whole-body leucine turnover, from which leucine rate of appearance (LRa, an index of protein breakdown) and leucine oxidation (Lox, a measure of irreversible protein loss) were estimated, energy expenditure and substrate utilization. We measured the concentration of liver transaminases as well as of testosterone, SHBG and IGF1. Results: Testosterone treatment significantly reduced LRa by 7.1G2.5% and Lox by 14.6G4.5% (P!0.05). The concentration of liver transaminases did not change significantly, while that of serum SHBG fell within the normal range by 16.8G4.0% and that of IGF1 increased by 18.4G7.7% (P!0.05). The concentration of peripheral testosterone increased from 0.4G0.1 to 1.1G0.2 nmol/l (P!0.05), without exceeding the upper normal limit. There was no change in energy expenditure and fat and carbohydrate utilization. Conclusions: Hepatic exposure to unesterified testosterone by oral delivery stimulates protein anabolism by reducing protein breakdown and oxidation without inducing systemic androgen excess in women. We conclude that a small oral dose of unesterified testosterone holds promise as a simple novel treatment of protein catabolism and muscle wasting.

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“…Recovery was determined by comparing four replicate injections samples spiked with standards in low and high levels. Carry-over was evaluated when time of analysis is greater and much more laborious than our method [13,14].…”

Section: Assay Validationmentioning

confidence: 99%

Analytical Performance of LC-MS/MS Method for Simultaneous Determination of Five Steroids in Serum

Alves¹

2015

Mass Spectrom Purif Tech

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The accurate measurement of steroid hormones is the fulcrum of modern endocrinology. Our aim was to develop a mass spectrometry coupled with a liquid chromatographic method for 17-hydroxyprogesterone, androstenedione, 11-desoxycortisol, 21-desoxycortisol and cortisol serum quantification simultaneously. The samples were submitted to isotopic internal standards addition, protein precipitation and two dimensional liquid chromatography consisting of trapping column and reverse-phase C18 analytical column following atmospheric pressure chemical ionization and mass spectrometry detection. For all compounds tested functional sensitivity was less than 0.5 ng/mL, precision was less than 15%, recovery ranged from 93% to 120%, linearity ranged from 89% to 111%, and accuracy was considered adequate. In conclusion, we developed a suitable, fast and reliable method for simultaneous routine measurement of 17-hydroxyprogesterone, androstenedione, 11-desoxycortisol, 21-desoxycortisol and cortisol in serum.

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Development and validation of a sensitive liquid chromatography–tandem mass spectrometry assay to simultaneously measure androgens and estrogens in serum without derivatization

Cited by 224 publications

References 43 publications

Effect of Testosterone Treatment on Glucose Metabolism in Men With Type 2 Diabetes: A Randomized Controlled Trial

Effect of Testosterone Treatment on Glucose Metabolism in Men With Type 2 Diabetes: A Randomized Controlled Trial

Oral low-dose testosterone administration induces whole-body protein anabolism in postmenopausal women: a novel liver-targeted therapy

Analytical Performance of LC-MS/MS Method for Simultaneous Determination of Five Steroids in Serum

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