Age- and sex-adjusted reference intervals for steroid hormones measured by liquid chromatography–tandem mass spectrometry using a widely available kit
Open accessAbstract
Objective: Measurements utilizing commercially available sets of reagents for determination of steroid hormone profiles by liquid chromatography-tandem mass spectrometry (LC-MS/MS) have become increasingly important for routine laboratories. However, method-specific publications of reference intervals obtained from sufficiently large studies are often missing. Methods: After validation of performance characteristics, a widely available kit for steroid analysis by LC-MS/MS was used to measure concentrations of 15 endogenous steroids (aldosterone, cortisol, cortisone, corticosterone, 11-deoxycortisol, 21-deoxycortisol, dehydroepiandrosterone sulfate, estradiol, testosterone, androstenedione, dihydrotestosterone, dehydroepiandrosterone, 17-hydroxyprogesterone, 11-deoxycorticosterone, progesterone) in more than 500 blood samples from a population-based study. While randomly selected from a larger cohort, the samples equally represented both sexes and covered a wide range of adult age groups. Age- and sex-specific reference intervals were calculated, and correlation with BMI was assessed. Results: Performance characteristics of the assay matched expectations for 9 of 15 steroids. For most of them, reference intervals obtained from our study population were comparable to those reported by others, with age and sex being the major determinants. A sex-specific correlation with BMI was found for seven steroids. We identified limitations regarding sensitivity of the method for quantification of progesterone in males and postmenopausal females. Concentrations of aldosterone, 21-deoxycortisol, estradiol, 11-deoxycorticosterone, and dihydrotestosterone could not be quantified in a large percentage of samples. Conclusions: The reference intervals for nine steroids will support meaningful interpretation for steroid profiles as measured by a widely used kit for LC-MS/MS-based quantification. Laboratories using such kits must be aware of potential limitations in sensitivity for some steroids included in the profile. Significance Statement: Quantification of steroid hormones is a cornerstone for diagnosis of several diseases. Commonly used immunoassays have limitations in specificity. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is a promising alternative, particularly if methods are harmonized across laboratories. The use of kits from commercial suppliers might support this. Clinical interpretation of steroid concentrations requires availability of appropriate reference intervals (RIs), but studies on RIs reported in the literature differ in preanalytical and analytical procedures. Here, we provide RIs for steroids measured by a widely available kit under preanalytical conditions mirroring common clinical practice. Such RIs might facilitate interpretation for those using the same method and comparable conditions in clinical routine.
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Several sensitive and robust LC-MS/MS methods for multiple steroids have been developed in recent years and many methods meet the requirements for clinical applications.
“Several sensitive and robust LC-MS/MS methods for multiple steroids have been developed in recent years and many methods meet the re quirements for clinical applications [5 -16].”
From Sex- and age-specific reference intervals of 16 steroid metabolites quantified simultaneously by LC-MS/MS in sera from 2458 healthy subjects aged 0 to 77 years (2024) · cites this paper as [14]
The Introduction describes a commercially available LC-MS/MS kit for measuring 15 steroids, and the Conclusion says it performs adequately for some steroids. This supports the paper as an example of a multi-steroid method, but it does not establish that many methods meet clinical requirements. The Discussion also reports limited sensitivity for six steroids in the kit; page numbers were not available in our copy. (LLM, conf=0.84)
Experimental — accuracy not yet measuredPreviously published methods including multiple steroid metabolites used larger sample volumes (150-500 μL) and/or longer total run-times (13.5-17 min).
“Previously published methods including multiple steroid metabolites used either larger sample volumes from 150-500 µ L and/or longer total rum-times from 13.5-17 min. [5,7,9,14,22].”
From Sex- and age-specific reference intervals of 16 steroid metabolites quantified simultaneously by LC-MS/MS in sera from 2458 healthy subjects aged 0 to 77 years (2024) · cites this paper as [14]
The Materials and methods section (Laboratory measurements; page unavailable) describes a multi-steroid assay using 500 µL of sample, supporting one endpoint of the stated volume range. The passages do not establish a 150 µL method or a total run time of 13.5–17 minutes for this method. (LLM, conf=0.86)
Experimental — accuracy not yet measuredReference intervals have been published for five to nine adrenal and androgenic steroids in men and women.
“and five to nine adrenal and androgenic steroids in men and women [8,13,14,21]”
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Cites
2016 European Guidelines on cardiovascular disease prevention in clinical practice
Piepoli, M. F., Hoes, A. W., Agewall, S., Albus, C., Brotons, C., Catapano, A. L., Cooney, M.-T., Corrà, U., Cosyns, B., Deaton, C., et al. (2016) · European Heart Journal · doi:10.1093/eurheartj/ehw106 · Cited by 6,457 · see citing works
Validation of bioanalytical LC–MS/MS assays: Evaluation of matrix effects
Eeckhaut, A. V., Lanckmans, K., Sarre, S., Smolders, I. J., & Michotte, Y. M. (2009) · Journal of Chromatography B · doi:10.1016/j.jchromb.2009.01.003 · Cited by 763 · see citing works
KORA - A Research Platform for Population Based Health Research
Holle, R., Happich, M., Löwel, H., & Wichmann, H. ‐. (2005) · Das Gesundheitswesen · doi:10.1055/s-2005-858235 · Cited by 753 · see citing works
Assessing obesity: classification and epidemiology
Seidell, J. C., & Flegal, K. M. (1997) · British Medical Bulletin · doi:10.1093/oxfordjournals.bmb.a011611 · Cited by 499 · see citing works
Handelsman, D. J., & Wartofsky, L. (2013) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jc.2013-3375 · Cited by 368 · see citing works
European Medicines Agency Guideline on Bioanalytical Method Validation: What More Is There to Say?
Smith, G. (2012) · Bioanalysis · doi:10.4155/bio.12.44 · Cited by 308 · see citing works
Wudy, S. A., Schuler, G., Sánchez-Guijo, A., & Hartmann, M. F. (2018) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2017.09.003 · Cited by 236 · see citing works
Determination of cortisol in serum, saliva and urine
Turpeinen, U., & Hämäläinen, E. (2013) · Best Practice & Research Clinical Endocrinology & Metabolism · doi:10.1016/j.beem.2013.10.008 · Cited by 214 · see citing works
Royston, P. J., & Wright, E. M. (1998) · Journal of the Royal Statistical Society Series A (Statistics in Society) · doi:10.1111/1467-985x.00091 · Cited by 195 · see citing works
Haring, R., Hannemann, A., John, U., Radke, D., Nauck, M., Wallaschofski, H., Owen, L., Adaway, J., Keevil, B. G., & Brabant, G. (2012) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jc.2011-2134 · Cited by 185 · see citing works
Osuna C., J. A., Gómez-Pérez, R., Arata-Bellabarba, G., & Villaroel, V. (2006) · Archives of Andrology · doi:10.1080/01485010600692017 · Cited by 183 · see citing works
Kuan, V., Denaxas, S., Patalay, P., Nitsch, D., Mathur, R., González-Izquierdo, A., Sofat, R., Partridge, L., Roberts, A., Wong, I. C. K., et al. (2022) · The Lancet Digital Health · doi:10.1016/s2589-7500(22)00187-x · Cited by 183 · see citing works
Vogeser, M., & Parhofer, K. G. (2007) · Experimental and Clinical Endocrinology & Diabetes · doi:10.1055/s-2007-981458 · Cited by 177 · see citing works
Eisenhofer, G., Peitzsch, M., Kaden, D., Langton, K., Pamporaki, C., Masjkur, J., Tsatsaronis, G., Mangelis, A., Williams, T. A., Reincke, M., et al. (2017) · Clinica Chimica Acta · doi:10.1016/j.cca.2017.05.002 · Cited by 171 · see citing works
Self-citationTraceability in Laboratory Medicine
Vesper, H. W., & Thienpont, L. M. (2009) · Clinical Chemistry · doi:10.1373/clinchem.2008.107052 · Cited by 169 · see citing works
Bae, Y. J., Zeidler, R., Baber, R., Vogel, M., Wirkner, K., Loeffler, M., Ceglarek, U., Kiess, W., Körner, A., Thiery, J., et al. (2019) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2019.105409 · Cited by 123 · see citing works
Vesper, H. W., Myers, G. L., & Miller, W. G. (2016) · American Journal of Clinical Nutrition · doi:10.3945/ajcn.115.110387 · Cited by 112 · see citing works
Health Problems in Congenital Adrenal Hyperplasia due to 21-Hydroxylase Deficiency
Reisch, N., Arlt, W., & Krone, N. P. (2011) · Hormone Research in Paediatrics · doi:10.1159/000327794 · Cited by 107 · see citing works
Body fatness and sex steroid hormone concentrations in US men: results from NHANES III
Rohrmann, S., Shiels, M. S., López, D. S., Rifai, N., Nelson, W. G., Kanarek, N., Güallar, E., Menke, A., Joshu, C. E., Feinleib, M., et al. (2011) · Cancer Causes & Control · doi:10.1007/s10552-011-9790-z · Cited by 105 · see citing works
Hannemann, A., Bidlingmaier, M., Friedrich, N., Manolopoulou, J., Spyroglou, A., Völzke, H., Beuschlein, F., Seißler, J., Rettig, R., Felix, S. B., et al. (2012) · European Journal of Endocrinology · doi:10.1530/eje-11-1013 · Cited by 103 · see citing works
Self-citationPlasma Steroid Metabolome Profiling for Diagnosis and Subtyping Patients with Cushing Syndrome
Eisenhofer, G., Masjkur, J. R., Peitzsch, M., Dalmazi, G. D., Bidlingmaier, M., Grüber, M., Fazel, J., Oßwald, A., Beuschlein, F., & Reincke, M. (2017) · Clinical Chemistry · doi:10.1373/clinchem.2017.282582 · Cited by 99 · see citing works
Self-citationMultimorbidität und erfolgreiches Altern
Peters, A., Döring, A., Ladwig, K., Meisinger, C., Linkohr, B., Autenrieth, C. S., Baumeister, S., Behr, J., Bergner, A., Bickel, H., et al. (2011) · Zeitschrift für Gerontologie und Geriatrie · doi:10.1007/s00391-011-0245-7 · Cited by 96 · see citing works
Self-citationKhamis, M. M., Adamko, D. J., & El‐Aneed, A. (2019) · Mass Spectrometry Reviews · doi:10.1002/mas.21607 · Cited by 89 · see citing works
Verdonk, S. J. E., Vesper, H. W., Martens, F., Sluss, P. M., Hillebrand, J. J., & Heijboer, A. C. (2019) · Clinica Chimica Acta · doi:10.1016/j.cca.2019.04.062 · Cited by 83 · see citing works
Maggio, M., Lauretani, F., Basaria, S., Ceda, G. P., Bandinelli, S., Metter, E. J., Bos, A. J., Ruggiero, C., Ceresini, G., Paolisso, G., et al. (2008) · Journal of Endocrinological Investigation · doi:10.1007/bf03345608 · Cited by 77 · see citing works
Primary aldosteronism: current knowledge and controversies in Conn's syndrome
Schirpenbach, C., & Reincke, M. (2007) · Nature Clinical Practice Endocrinology & Metabolism · doi:10.1038/ncpendmet0430 · Cited by 70 · see citing works
Self-citationBui, H. N., Sluss, P. M., Hayes, F. J., Blincko, S. J., Knol, D. L., Blankenstein, M. A., & Heijboer, A. C. (2015) · Clinica Chimica Acta · doi:10.1016/j.cca.2015.08.019 · Cited by 64 · see citing works
Parikh, T. P., Stolze, B. R., Özarda, Y., Jonklaas, J., Welsh, K. J., Masika, L., Hill, M. J., DeCherney, A. H., & Soldin, S. J. (2018) · Endocrine Connections · doi:10.1530/ec-18-0417 · Cited by 51 · see citing works
Few European Adults are Living a Healthy Lifestyle
Marques, A., Peralta, M., Martins, J., Loureiro, V., Almanzar, P. C., & de Matos, M. G. (2018) · American Journal of Health Promotion · doi:10.1177/0890117118787078 · Cited by 44 · see citing works
Owen, L. J., & Keevil, B. G. (2012) · Annals of Clinical Biochemistry International Journal of Laboratory Medicine · doi:10.1258/acb.2012.012037 · Cited by 42 · see citing works
Mezzullo, M., Pelusi, C., Fazzini, A., Repaci, A., Di Dalmazi, G., Gambineri, A., Pagotto, U., & Fanelli, F. (2020) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2019.105538 · Cited by 39 · see citing works
Franscini, L. C., Vazquez-Montes, M., Buclin, T., Perera, R., Dunand, M., Grouzmann, E., & Popovic, M. B. (2015) · Pediatric Blood & Cancer · doi:10.1002/pbc.25385 · Cited by 38 · see citing works
Fanelli, F., Cantù, M., Temchenko, A., Mezzullo, M., Lindner, J. M., Peitzsch, M., Hawley, J. M., Bruce, S., Binz, P.-A., Ackermans, M. T., et al. (2022) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm-2021-1028 · Cited by 36 · see citing works
Diagnosis of Diseases of Steroid Hormone Production, Metabolism and Action
Honour, J. W. (2009) · Journal of Clinical Research in Pediatric Endocrinology · doi:10.4274/jcrpe.v1i5.209 · Cited by 30 · see citing works
Systematic review of serum steroid reference intervals developed using mass spectrometry
Tavita, N., & Greaves, R. F. (2017) · Clinical Biochemistry · doi:10.1016/j.clinbiochem.2017.07.002 · Cited by 29 · see citing works
Yasmin, E., Balen, A. H., & Barth, J. H. (2012) · European Journal of Obstetrics & Gynecology and Reproductive Biology · doi:10.1016/j.ejogrb.2012.09.025 · Cited by 27 · see citing works
Emeny, R. T., Bidlingmaier, M., Lacruz, M. E., Linkohr, B., Peters, A., Reincke, M., & Ladwig, K. (2014) · Experimental Gerontology · doi:10.1016/j.exger.2014.08.001 · Cited by 20 · see citing works
Self-citationAdaway, J., Keevil, B. G., Miller, A. M., Monaghan, P. J., Merrett, N., & Owen, L. J. (2019) · Annals of Clinical Biochemistry International Journal of Laboratory Medicine · doi:10.1177/0004563219888549 · Cited by 16 · see citing works
Wilkinson, I. B., Raine, T., Wiles, K., Goodhart, A. K., Hall, C., & O’Neill, H. (2017) · doi:10.1093/med/9780199689903.003.0017 · Cited by 4 · see citing works
Flagged first, then by citations
Research works printed but not linked: 2
- [12]NCCLS. How to define and determine reference intervals in the Clinical Laboratory; approved guideline—second edition. In NCCLS Document C28-A2, vol 20, pp 1– 38. Wayne, PA, USA: National Committee for Clinical Laboratory Standards, 2000.
- [27]Plebani M. Quality indicators to detect pre-analytical errors in laboratory testing. Clinical Biochemist. Reviews 2012 33 85– 88. (10.1016/j.cca.2013.07.033) 22930602 PMC3428256 · DOI
Cited by
Sorted by citations
The Glucocorticoid Receptor: Isoforms, Functions, and Contribution to Glucocorticoid Sensitivity
Lockett, J., Inder, W. J., & Clifton, V. L. (2024) · Endocrine Reviews · doi:10.1210/endrev/bnae008 · Cited by 135 · see citing works
Frederiksen, H., Johannsen, T. H., Andersen, S. E., Petersen, J. H., Busch, A. S., Ljubicic, M. L., Fischer, M. B., Upners, E. N., Hagen, C. P., Main, K. M., et al. (2024) · Clinica Chimica Acta · doi:10.1016/j.cca.2024.119852 · Cited by 46 · see citing works
- Authors
- Kunz, S., Wang, X., Ferrari, U., Drey, M., Theodoropoulou, M., Schilbach, K., Reincke, M., Heier, M., Peters, A., Koenig, W., Zeller, T., Thorand, B., & Bidlingmaier, M.
- Affiliations
- Ludwig-Maximilians-Universität München (DE)
- Department of Medicine IV, University Hospital, Ludwig Maximilian University Munich, Munich, Germany
- University Hospital Augsburg (DE)
- Helmholtz Munich (DE)
- German Centre for Cardiovascular Research (DZHK), Partner site Munich Heart Alliance, Munich, Germany
- Universität Hamburg (DE)
- Corresponding author
- Bidlingmaier, Martin
- ORCID
- Venue
- Endocrine Connections
- Publisher
- Bioscientifica
- Published
- 2023-11-08
- Volume
- 13
- Issue
- 1
- ISSN
- 2049-3614 (ISSN-L)
- PMC
- PMC10762560
- OpenAlex
- W4388489371
- Cited by
- 14
- References
- 39
- Citations by year
- 2026: 6 · 2025: 3 · 2024: 4 · 2023: 1
- FWCI
- 2.49
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Endocrine Connections (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- PuSH - Publication Server of Helmholtz Zentrum München (submittedVersion, cc-by-nc)
- Funders
- Deutsches Zentrum für Herz-Kreislaufforschung
- European Commission (HORIZON2020, 694913)
- Deutsche Forschungsgemeinschaft (314061271, BMBF FKZ 01ET0713)
- Bundesministerium für Bildung und Forschung (BMBF FKZ 01ET0713, FKZ 01ET0713, 01ET1003A, 01ET0713)
- Helmholtz Zentrum München (FKZ 01ET0713)
- Topics
- Health Sciences › Medicine › Physiology › Clinical Laboratory Practices and Quality Control
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Hormonal and reproductive studies
- Keywords
- endogenous steroid hormones, liquid chromatography-tandem mass spectrometry, commercial assay kits, assay performance, body mass index, aldosterone, cortisol, estradiol, testosterone, dihydrotestosterone, dehydroepiandrosterone sulfate, 17-hydroxyprogesterone, deoxycorticosterone, 21-deoxycortisol, 11-deoxycortisol
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