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      Improving Science by Overcoming Laboratory Pitfalls With Hormone Measurements

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          Abstract

          Despite all the effort taken, there is often surprisingly little attention paid to the hormone analyses involved in research studies. Thinking carefully about the quality of the hormone measurements in these studies is, however, of major importance, as this attention to methods may prevent false conclusions and inappropriate follow-up studies. We discuss issues regarding hormone measurements that one should consider, ideally prior to starting, or otherwise, as they arise during a scientific study: quality of the technique, expertise, matrices, timing and storage conditions, freeze-thaw cycles, lot-to-lot and day-to-day variation, analyses per batch or sample-wise, singlicate or duplicate measurements, combining methods, and standardization. This article and the examples mentioned herein aim to clarify the need to pay attention to the hormone analyses, and to help in making decisions. In addition, these examples help editors and reviewers of scientific journals to pay attention to the methods section in the submitted manuscripts and ask the right critical questions when needed.

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          Most cited references78

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          Accuracy of 6 routine 25-hydroxyvitamin D assays: influence of vitamin D binding protein concentration.

          Recent recognition of its broad pathophysiological importance has triggered an increased interest in 25-hydroxyvitamin D [25(OH)D]. By consequence, throughput in 25(OH)D testing has become an issue for clinical laboratories, and several automated assays for measurement of 25(OH)D are now available. The aim of this study was to test the accuracy and robustness of these assays by comparing their results to those of an isotope dilution/online solid-phase extraction liquid chromatography/tandem mass spectrometry (ID-XLC-MS/MS) method. We put specific focus on the influence of vitamin D-binding protein (DBP) by using samples with various concentrations of DBP. We used 5 automated assays (Architect, Centaur, iSYS, Liaison, and Elecsys), 1 RIA (Diasorin) preceded by extraction, and an ID-XLC-MS/MS method to measure 25(OH)D concentrations in plasma samples of 51 healthy individuals, 52 pregnant women, 50 hemodialysis patients, and 50 intensive care patients. Using ELISA, we also measured DBP concentrations in these samples. Most of the examined 25(OH)D assays showed significant deviations in 25(OH)D concentrations from those of the ID-XLC-MS/MS method. As expected, DBP concentrations were higher in samples of pregnant women and lower in samples of IC patients compared to healthy controls. In 4 of the 5 fully automated 25(OH)D assays, we observed an inverse relationship between DBP concentrations and deviations from the ID-XLC-MS/MS results. 25(OH)D measurements performed with most immunoassays suffer from inaccuracies that are DBP concentration dependent. Therefore, when interpreting results of 25(OH)D measurements, careful consideration of the measurement method is necessary.
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            Requirement for mass spectrometry sex steroid assays in the Journal of Clinical Endocrinology and Metabolism.

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              A Reappraisal of Testosterone's Binding in Circulation: Physiological and Clinical Implications.

              In the circulation, testosterone and other sex hormones are bound to binding proteins, which play an important role in regulating their transport, distribution, metabolism, and biological activity. According to the free hormone hypothesis, which has been debated extensively, only the unbound or free fraction is biologically active in target tissues. Consequently, accurate determination of the partitioning of testosterone between bound and free fractions is central to our understanding of how its delivery to the target tissues and biological activity are regulated and consequently to the diagnosis and treatment of androgen disorders in men and women. Here, we present a historical perspective on the evolution of our understanding of the binding of testosterone to circulating binding proteins. On the basis of an appraisal of the literature as well as experimental data, we show that the assumptions of stoichiometry, binding dynamics, and the affinity of the prevailing models of testosterone binding to sex hormone-binding globulin and human serum albumin are not supported by published experimental data and are most likely inaccurate. This review offers some guiding principles for the application of free testosterone measurements in the diagnosis and treatment of patients with androgen disorders. The growing number of testosterone prescriptions and widely recognized problems with the direct measurement as well as the computation of free testosterone concentrations render this critical review timely and clinically relevant.
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                Author and article information

                Journal
                J Clin Endocrinol Metab
                J Clin Endocrinol Metab
                jcem
                The Journal of Clinical Endocrinology and Metabolism
                Oxford University Press (US )
                0021-972X
                1945-7197
                April 2021
                31 December 2020
                31 December 2020
                : 106
                : 4
                : e1504-e1512
                Affiliations
                Amsterdam UMC, Vrije Universiteit Amsterdam and University of Amsterdam, Endocrine Laboratory, Department of Clinical Chemistry, Amsterdam Gastroenterology Endocrinology & Metabolism , Amsterdam, Netherlands
                Author notes
                Correspondence: Annemieke C. Heijboer, Amsterdam UMC, Endocrine Laboratory (K2-283), Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands. Email: a.heijboer@ 123456amsterdamumc.nl
                Author information
                http://orcid.org/0000-0002-6712-9955
                Article
                dgaa923
                10.1210/clinem/dgaa923
                7993596
                33382880
                c4406df2-bbad-4781-9ec2-ab698dfd44a7
                © The Author(s) 2020. Published by Oxford University Press on behalf of the Endocrine Society.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence ( http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

                History
                : 11 September 2020
                : 04 December 2020
                : 11 January 2020
                Page count
                Pages: 9
                Categories
                Online Only Articles
                Mini-Reviews
                AcademicSubjects/MED00250

                Endocrinology & Diabetes
                quality,immunoassay,mass spectrometry,matrix,technique,pre-analysis
                Endocrinology & Diabetes
                quality, immunoassay, mass spectrometry, matrix, technique, pre-analysis

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