Testosterone Measurement by Isotope-Dilution Liquid Chromatography–Tandem Mass Spectrometry: Validation of a Method for Routine Clinical Practice

Cawood, Marion L.; Field, H P; Ford, Clive; Gillingwater, Scott; Kicman, Andrew T.; Cowan, David; Barth, Julian H.

doi:10.1373/clinchem.2004.044503

Cited by 138 publications

(83 citation statements)

References 15 publications

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“…A chromatogram of Te extracted from the serum of a healthy woman is shown in Figure 6, and the method's performance characteristics are listed in Table 2. Considering the analytical sensitivity and small volume of sample used (100 mL), this method is one of the most sensitive of the published LC-MS/MS methods for the measurement of Te (Cawood et al, 2005;Vicente et al, 2006;Borrey et al, 2007;Thienpont et al, 2008). To address the issue of poor performance of commercial IAs for Te, it was suggested to use the isotope-dilution LC-MS/MS method for the routine measurement of Te in samples from women and children (Rosner et al, 2007).…”

Section: Androgensmentioning

confidence: 99%

Liquid chromatography-tandem mass spectrometry applications in endocrinology

Kushnir

Rockwood

Bergquist

2009

Mass Spectrom. Rev.

139

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Liquid chromatography-tandem mass spectrometry (LC-MS/ MS) has been recognized as a primary methodology for the accurate analysis of endogenous steroid hormones in biological samples. This review focuses on the use of LC-MS/MS in clinical laboratories to assist with the diagnosis of diverse groups of endocrine and metabolic diseases. Described analytical methods use on-line and off-line sample preparation and analytical derivatization to enhance analytical sensitivity, specificity, and clinical utility. Advantages of LC-MS/MS as an analytical technique include high specificity, possibility to simultaneously measure multiple analytes, and the ability to assess the specificity of the analysis in every sample. All described analytical methods were extensively validated, utilized in routine diagnostic practice, and were applied in a number of clinical and epidemiological studies, including a study of the steroidogenesis in ovarian follicles. #

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Section: Androgensmentioning

confidence: 99%

Liquid chromatography-tandem mass spectrometry applications in endocrinology

Kushnir

Rockwood

Bergquist

2009

Mass Spectrom. Rev.

139

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“…Radioimmunoassays (RIAs) are commonly used for the determination of the actual levels of testosterone (TS) (2). As RIA methods have the inherent limitations in specificity, it is difficult to determine accurately the low levels of TS present in females and children (3,4). Gas chromatography-mass spectrometry is used for the determination of low levels of natural steroids with excellent specificity and sensitivity (5).…”

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confidence: 99%

Accurate determination of tissue steroid hormones, precursors and conjugates in adult male rat

Maeda

Tanaka

Suzuki

et al. 2012

The Journal of Biochemistry

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The actual levels of steroid hormones in organs are vital for endocrine, reproductive and neuronal health and disorders. We developed an accurate method to determine the levels of steroid hormones and steroid conjugates in various organs by an efficient preparation using a solid-phase-extraction cartridge. Each steroid was identified by the precursor ion spectra using liquid chromatographyelectrospray ionization timeof-flight mass spectrometry, and the respective steroids were quantitatively analysed in the selected reaction monitoring mode by liquid chromatograph-mass spectrometry/mass spectrometry (LC-MS/MS). The data showed that significant levels of testosterone, corticosterone and precursors of both hormones were detected in all organs except liver. The glucuronide conjugates of steroid hormones and the precursors were detected in all organs except liver, but sulfate conjugates of these steroids were observed only in the target organs of the hormones and kidney. Interestingly, these steroids and the conjugates were not observed in the liver except pregnenolone. In conclusion, an accurate determination of tissue steroids was developed using LC-MS analysis. Biosynthesis of steroid hormones from the precursors was estimated even in the target organs, and the delivery of these steroid conjugates was also suggested via the circulation without any significant hepatic participation.

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“…Trata-se de técnica com alta especificidade e sensibilidade, sendo considerada referência para a dosagem de hormônios esteroides (31). No caso da testosterona, várias publicações fundamentadas em HPLC como método preparativo e espectrometria de massa em tandem para quantificação foram recentemente apresentadas e demonstraram sua superioridade em relação aos métodos tradicionais, especialmente em amostras de mulheres e crianças (32)(33)(34).…”

Section: Discussionunclassified

Testosterona falsamente elevada em uma paciente diabética tipo 1 com acne e insuficiência renal crônica em diálise

Marcondes

Martins

Amaral

et al. 2012

Arq Bras Endocrinol Metab

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SUMÁRIODescrevemos uma paciente com diabetes tipo 1 com acne e insuficiência renal crônica, em diálise, que apresentou uma dosagem de testosterona total e livre elevada (612 ng/dL, normal < 90 ng/dL e 255 pMol/L, normal: 20-45 pMol/L, respectivamente). Na avaliação clínica, além da acne facial não havia qualquer outro sinal de hiperandrogenismo. Após ter esse resultado confirmado, ela foi submetida à avaliação morfológica de adrenal e ovários (tomografia computadorizada de adrenal e ultrassom pélvico), cujos resultados foram normais. Na ausência de quadro clínico de virilização, foram consideradas outras possibilidades que pudessem explicar a elevação da testosterona, entre as quais a presença de comorbidades (diabetes e insuficiência renal crônica) e falha do método de dosagem. Uma nova determinação da testosterona total, por meio da cromatografia líquida de alta performance como método preparativo e espectrometria de massa em tandem, resultou normal (21 ng/dL), compatível com uma concentração de testosterona total falsamente elevada pela presença de interferentes com o método de dosagem. Arq Bras Endocrinol Metab. 2012;56(5):319-23 SUMMARYThe purpose of this report is to present the case of a patient with type 1 diabetes with acne and chronic renal failure on dialysis admitted to the hospital with high total total and free testosterone (612 ng/dL, normal < 90 ng/dL; 255 pMol/L, normal: 20-45 pMol/L). On clinical evalua tion, she presented facial acne, and no other signs of hyperandrogenism. As this result was confirmed, she underwent adrenal and ovary morphological assessment (adrenal CT and pelvic ultrasound), which yielded normal results. Due to divergence between clinical and laboratory findings, we considered other possibilities that could explain the elevation of testosterone, including the presence of comorbidities (diabetes and chronic renal failure) and failure of the testosterone assay. Testosterone levels were determined again by high performance liquid chromatography, as a preparative method, and tandem mass spectrometry, yielding normal results (21 ng/dL), which were compatible with a falsely elevated total testosterone level caused by the presence of factors that intereferred with the assay. INTRODUÇÃO A s síndromes hiperandrogênicas englobam doenças que se manifestam pelo aumento da atividade biológica dos andrógenos. Na mulher, essas manifestações incluem hirsutismo, acne, alopecia tipo androgêni-ca, disfunção menstrual, infertilidade, abortamento precoce e sinais de virilização (atrofia do parênquima mamário, alteração da tonalidade da voz, redistribuição de massas musculares e clitoromegalia).Tanto a testosterona total (forma livre e forma ligada às proteínas transportadoras) quanto a testosterona livre são consideradas marcadores sensíveis para a demonstração da hiperandrogenemia (1). Entretanto, não existe correlação entre a concentração dos andrógenos

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Testosterone Measurement by Isotope-Dilution Liquid Chromatography–Tandem Mass Spectrometry: Validation of a Method for Routine Clinical Practice

Cited by 138 publications

References 15 publications

Liquid chromatography-tandem mass spectrometry applications in endocrinology

Liquid chromatography-tandem mass spectrometry applications in endocrinology

Accurate determination of tissue steroid hormones, precursors and conjugates in adult male rat

Testosterona falsamente elevada em uma paciente diabética tipo 1 com acne e insuficiência renal crônica em diálise

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