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High sensitivity tandem mass spectrometry assay for serum estrone and estradiol
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry, Analytical Chemistry.
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2008 (English)In: American Journal of Clinical Pathology, ISSN 0002-9173, E-ISSN 1943-7722, Vol. 129, no 4, 530-539 p.Article in journal (Refereed) Published
Abstract [en]

High-sensitivity measurement of serum estrogens is important in adult and pediatric endocrinology and oncology. We developed a high-sensitivity liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for simultaneous measurement of estrone (E-1) and estradiol (E-2). Aliquots of 200 mu L of serum were spiked with internal standard, extracted, derivatized with dansyl chloride, and analyzed by LC-MS/MS using 2-dimensional chromatographic separation. Total imprecision for the method was less than 11%; the limit of quantitation was 1 pg/mL. Reference intervals were established with samples from more than 900 healthy postmenopausal women, men, girls, and boys. Concentrations of estrogens in children reached adult levels by Tanner stage 3. In men and postmenopausal women, the median concentrations of total estrogens (E-1 + E-2) were 39 and 22 pg/mL, and the median E-2/E-1 ratios were 0.98 and 0.55, respectively. The method requires a small sample volume and has adequatesensitivity and specificity for analyzing estrogens in samples from postmenopausal women, men, and children.

Place, publisher, year, edition, pages
2008. Vol. 129, no 4, 530-539 p.
National Category
Medical and Health Sciences
URN: urn:nbn:se:uu:diva-97094DOI: 10.1309/LC03BHQ5XJPJYEKGISI: 000254127600003OAI: oai:DiVA.org:uu-97094DiVA: diva2:171883
Available from: 2008-04-22 Created: 2008-04-22 Last updated: 2013-03-06Bibliographically approved
In thesis
1. Mass Spectrometric Applications for Diagnosing Metabolic and Endocrine Diseases
Open this publication in new window or tab >>Mass Spectrometric Applications for Diagnosing Metabolic and Endocrine Diseases
2008 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Disease-specific compounds (biomarkers) are analyzed in clinical laboratories to assist with diagnosing diseases. This thesis describes development and validation of liquid chromatography tandem mass spectrometry (LC-MS/MS) based tests for diagnosing a diverse group of endocrine and metabolic diseases. The analytical methods used on-line and off-line sample extraction and analytical derivatization as means of enhancing the analytical sensitivity, specificity and clinical utility. All developed methods were extensively validated and reference intervals for the biomarker concentrations were established in blood samples of healthy adults and children. Advantages of the LC-MS/MS as an analytical technique include possibility of simultaneous measurement of multiple analytes and ability of confirming their identity. In this thesis we proposed and evaluated approaches for the assessment of the specificity of analysis in the methods that use tandem mass spectrometry detection. To enhance throughput of the LC-MS/MS tests for the biomarkers that have endogenous or exogenous isomers an approach was developed for quantitation of isomers from unresolved chromatographic peaks. Using methods developed in this thesis we performed a study of the steroidogenesis in ovarian follicles of healthy women and women with polycystic ovary syndrome (PCOS). Obtained data on the steroid concentrations and associations between the steroid metabolites in the pathway would be helpful for better understanding of the ovarian pathophysiology. Potential biomarkers of PCOS were identified in the thesis; further studies will be necessary to confirm their clinical utility.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2008. 80 p.
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 426
Analytical chemistry, biomarker, liquid chromatography, tandem mass spectrometry, LC-MS/MS, sample preparation, isomers, steroid hormones, congenital adrenal hyperplasia, polycystic ovary syndrome, Analytisk kemi
urn:nbn:se:uu:diva-8658 (URN)978-91-554-7172-9 (ISBN)
Public defence
2008-05-16, BMC B:41, BMC, Husargatan 3, SE-751 24, Uppsala, Sweden, 10:15
Available from: 2008-04-22 Created: 2008-04-22 Last updated: 2011-05-10Bibliographically approved

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