Measurement of Serum Testosterone in Nondiabetic Young Obese Men: Comparison of Direct Immunoassay to Liquid Chromatography-Tandem Mass Spectrometry
Open accessAbstract
Hypoandrogenemia, a frequent finding in men with obesity, is defined by low concentrations of serum testosterone. Although immunoassay (IA) is the most used method for the determination of this steroid in clinical practice, liquid chromatography-mass spectrometry (LC-MS/MS) is considered a more reliable method. In this study, we aimed to compare IA versus LC-MS/MS measurement for the diagnosis of hypoandrogenemia in a cohort of 273 nondiabetic young obese men. Mean total testosterone (TT) levels were 3.20 ± 1.24 ng/mL for IA and 3.78 ± 1.4 ng/mL for LC-MS/MS. 53.7% and 26.3% of patients were classified as presenting hypoandrogenemia with IA and LC-MS/MS, respectively. Considering LC-MS/MS as the reference method, sensitivity and specificity of IA were 91.4% (95% CI 82.3–96.8) and 61.1% (95% CI 54.0–67.8), respectively. IA presented an AUC of 0.879 (95% CI 0.83–0.928). Multivariate regression analysis indicated that sex hormone-binding globulin (SHBG) concentrations (p = 0.002) and insulin resistance (p = 0.008) were factors associated with discrepant IA values. In conclusion, the determination of TT by IA in nondiabetic young men with obesity yields lower concentrations of TT than LC-MS/MS, resulting in an equivocal increased diagnosis of hypoandrogenemia, which could lead to inaccurate diagnosis and unnecessary treatment.
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Liquid chromatography-mass spectrometry is well accepted as the preferred technique for the analysis of testosterone.
“Liquid chromatography–mass spectrometry is well accepted as the preferred technique for the analysis of testosterone [90, 91].”
From Integrating Transwomen and Female Athletes with Differences of Sex Development (DSD) into Elite Competition: The FIMS 2021 Consensus Statement (2021) · cites this paper as [91]
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Cites
Matthews, D. R., Hosker, J. P., Rudenski, A. S., Naylor, B. A., Treacher, D. F., & Turner, R. C. (1985) · Diabetologia · doi:10.1007/bf00280883 · Cited by 31,502 · cited 1× · see citing works
“We used the homeostatic model assessment of insulin resistance (HOMA-IR) to determine the status of insulin resistance [22].”
MethodA Critical Evaluation of Simple Methods for the Estimation of Free Testosterone in Serum
Vermeulen, A., Verdonck, L., & Kaufman, J. M. (1999) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jcem.84.10.6079 · Cited by 3,824 · see citing works
Feldman, H. A., Longcope, C., Derby, C. A., Johannes, C. B., Araujo, A. B., Coviello, A. D., Bremner, W. J., & McKinlay, J. B. (2002) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jcem.87.2.8201 · Cited by 1,861 · see citing works
Testosterone Therapy in Men With Hypogonadism: An Endocrine Society Clinical Practice Guideline.
Bhasin, S., Brito, J. P., Cunningham, G. R., Hayes, F. J., Hodis, H. N., Matsumoto, A. M., Snyder, P. J., Swerdloff, R. S., Wu, F. C., & Yialamas, M. A. (2018) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jc.2018-00229 · Cited by 1,736 · cited 4× · see citing works
“The rationale for this was that no differentiated cut-offs values for both techniques are available and, in addition, most clinical guidelines use testosterone cut-off values derived from LC-MS/MS assays [8].”
BackgroundMethodInfluentialRosner, W., Auchus, R. J., Azziz, R., Sluss, P. M., & Raff, H. (2007) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jc.2006-1864 · Cited by 1,176 · see citing works
Taieb, J., Mathian, B., Millot, F., Patricot, M. C., Mathieu, E., Queyrel, N., Lacroix, I., Somma-Delpéro, C., & Boudou, P. (2003) · Clinical Chemistry · doi:10.1373/49.8.1381 · Cited by 638 · see citing works
Wang, C., Catlin, D. H., Demers, L. M., Starcevic, B., & Swerdloff, R. S. (2004) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jc.2003-031287 · Cited by 580 · see citing works
Díz, P., Otero-González, A., Rodríguez‐Álvarez, M. X., Gudé, F., López, F. J. G., de Francisco, A., & González‐Quintela, A. (2013) · BMC Endocrine Disorders · doi:10.1186/1472-6823-13-47 · Cited by 554 · 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
Bhasin, S., Pencina, M., Jasuja, G. K., Travison, T. G., Coviello, A., Orwoll, E., Wang, P. Y., Nielson, C., Wu, F., Tajar, A., et al. (2011) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jc.2010-3012 · Cited by 355 · see citing works
Testosterone concentrations in diabetic and nondiabetic obese men
Dhindsa, S., Miller, M. G., McWhirter, C. L., Mager, D. E., Ghanim, H., Chaudhuri, A., & Dandona, P. (2010) · Diabetes Care · doi:10.2337/dc09-1649 · Cited by 352 · see citing works
Mass spectrometry and immunoassay: how to measure steroid hormones today and tomorrow
Taylor, A. E., Keevil, B., & Huhtaniemi, I. T. (2015) · European Journal of Endocrinology · doi:10.1530/eje-15-0338 · Cited by 320 · see citing works
Advantages and challenges of mass spectrometry assays for steroid hormones
Stanczyk, F. Z., & Clarke, N. J. (2010) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2010.05.001 · Cited by 265 · see citing works
Sikaris, K., McLachlan, R. I., Kazlauskas, R., de Kretser, D., Holden, C. A., & Handelsman, D. J. (2005) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jc.2005-0962 · Cited by 259 · see citing works
Huhtaniemi, I. T., Tajar, A., Lee, D. M., O’Neill, T. W., Finn, J. D., Bártfai, G., Boonen, S., Casanueva, F. F., Giwercman, A., Han, T. S., et al. (2012) · European Journal of Endocrinology · doi:10.1530/eje-11-1051 · Cited by 200 · see citing works
Zarotsky, V., Huang, M., Carman, W. J., Morgentaler, A., Singhal, P. K., Coffin, D., & Jones, T. H. (2014) · Andrology · doi:10.1111/andr.274 · Cited by 174 · see citing works
Testosterone replacement therapy: for whom, when and how?
Tsametis, C. P., & Isidori, A. M. (2018) · Metabolism · doi:10.1016/j.metabol.2018.03.007 · Cited by 96 · see citing works
Free Testosterone: Pumping up the Tires or Ending the Free Ride?
Handelsman, D. J. (2017) · Endocrine Reviews · doi:10.1210/er.2017-00171 · Cited by 78 · see citing works
Measuring testosterone in women and men
Kanakis, G., Tsametis, C. P., & Goulis, D. G. (2019) · Maturitas · doi:10.1016/j.maturitas.2019.04.203 · Cited by 78 · see citing works
Janse, F., Eijkemans, M. J. C., Goverde, A. J., Lentjes, E. G. W. M., Hoek, A., Lambalk, C. B., Hickey, T. E., Fauser, B. C. J. M., & Norman, R. J. (2011) · European Journal of Endocrinology · doi:10.1530/eje-11-0482 · Cited by 75 · see citing works
Miller, K. K. (2001) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jcem.86.6.7610 · Cited by 70 · see citing works
Hackbarth, J. S., Hoyne, J. B., Grebe, S. K. G., & Singh, R. (2010) · Steroids · doi:10.1016/j.steroids.2010.08.008 · Cited by 62 · see citing works
Mass spectrometry, immunoassay and valid steroid measurements in reproductive medicine and science
Handelsman, D. J. (2017) · Human Reproduction · doi:10.1093/humrep/dex078 · Cited by 58 · see citing works
Lamm, S. H., Chidakel, A. R., & Bansal, R. (2016) · Urologic Clinics of North America · doi:10.1016/j.ucl.2016.01.005 · Cited by 51 · see citing works
Secondary male hypogonadism: A prevalent but overlooked comorbidity of obesity
Fernandez-Garcia, J.-C., Molina‐Vega, M., Muñoz-Garach, A., Damas-Fuentes, M., & Tinahones, F. J. (2018) · Asian Journal of Andrology · doi:10.4103/aja.aja_44_18 · Cited by 45 · see citing works
Self-citationAndrogen deficiency in women; role of accurate testosterone measurements
Demers, L. M. (2010) · Maturitas · doi:10.1016/j.maturitas.2010.04.019 · Cited by 40 · see citing works
How low can you go? Analytical performance of five automated testosterone immunoassays
La'ulu, S. L., Kalp, K. J., & Straseski, J. A. (2018) · Clinical Biochemistry · doi:10.1016/j.clinbiochem.2018.05.008 · Cited by 37 · see citing works
Cao, Z. T., Botelho, J. C., Rej, R., & Vesper, H. W. (2017) · Clinica Chimica Acta · doi:10.1016/j.cca.2017.03.010 · Cited by 36 · see citing works
Mechanisms of Hypoandrogenemia in Healthy Aging Men
Liu, P. Y., Iranmanesh, A., Nehra, A. X., Keenan, D. M., & Veldhuis, J. D. (2005) · Endocrinology and Metabolism Clinics of North America · doi:10.1016/j.ecl.2005.07.008 · Cited by 34 · see citing works
Molina‐Vega, M., Asenjo‐Plaza, M., García‐Ruiz, M. C., Varea‐Marineto, E., Casal‐Nievas, N., Álvarez‐Millán, J. J., Cabezas-Sánchez, P., Cardona, F., Queipo‐Ortuño, M. I., Castellano‐Castillo, D., et al. (2019) · Obesity · doi:10.1002/oby.22579 · Cited by 23 · see citing works
Self-citationMontagna, G., Balestra, S., D’Aurizio, F., Romanelli, F., Benagli, C., Tozzoli, R., Risch, L., Giovanella, L., & Imperiali, M. (2018) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm-2017-1201 · Cited by 17 · see citing works
McCartney, C. R., Burt Solorzano, C. M., Patrie, J. T., Marshall, J. C., & Haisenleder, D. J. (2018) · Steroids · doi:10.1016/j.steroids.2018.09.001 · Cited by 9 · see citing works
Comas, I., Ferrer, R., Planas, J., Celma, A., Regis, L., & Moróte, J. (2017) · Actas Urológicas Españolas · doi:10.1016/j.acuro.2017.01.007 · Cited by 6 · see citing works
Comas, I., Ferrer, R., Planas, J., Celma, A., Regis, L., & Moróte, J. (2018) · Actas Urológicas Españolas (English Edition) · doi:10.1016/j.acuroe.2017.12.003 · Cited by 3 · see citing works
Cabarkapa, S., Perera, M., Sikaris, K. A., O'Brien, J. S., Bolton, D. M., & Lawrentschuk, N. (2017) · Prostate International · doi:10.1016/j.prnil.2017.05.003 · Cited by 2 · see citing works
Flagged first, then by citations
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Castelli, F. A., Rosati, G., Moguet, C., Fuentes‐Chust, C., Marrugo-Ramírez, J., Lefèbvre, T., Volland, H., Merkoçi, A., Simon, S., Fenaille, F., et al. (2021) · Analytical and Bioanalytical Chemistry · doi:10.1007/s00216-021-03586-z · Cited by 127 · see citing works
Saliva as Biomarker for Oral and Chronic Degenerative Non-Communicable Diseases
Basilicata, M., Pieri, M., Marrone, G., Nicolai, E., Lauro, M. D., Paolino, V., Tomassetti, F., Vivarini, I., Bollero, P., Bernardini, S., et al. (2023) · Metabolites · doi:10.3390/metabo13080889 · Cited by 35 · see citing works
Hamilton, B. R., Lima, G., Barrett, J., Seal, L., Kolliari-Turner, A., Wang, G., Karanikolou, A., Bigard, X., Löllgen, H., Zupet, P., et al. (2021) · Sports Medicine · doi:10.1007/s40279-021-01451-8 · Cited by 27 · see citing works
Fernández‐García, J. C., Barrios‐Rodríguez, R., Asenjo‐Plaza, M., Ramos‐Molina, B., Molina‐Vega, M., Guzmán-Guzmán, A., Moreno-León, L., Yubero‐Serrano, E. M., Ríus, F., Valdés, S., et al. (2022) · Metabolism · doi:10.1016/j.metabol.2022.155290 · Cited by 25 · see citing works
Self-citationNarinx, N., Nyamaah, J. A., David, K., Sommers, V., Walravens, J., Fiers, T., Lapauw, B., Decallonne, B., Claessens, F., Uytfanghe, K. V., et al. (2025) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm-2024-1237 · Cited by 10 · see citing works
Ohlsson, C., Li, L., Horkeby, K., Lawenius, L., Colldén, H., Sjögren, K., Baldanzi, G., Engström, G., Ärnlöv, J., Orho‐Melander, M., et al. (2025) · EBioMedicine · doi:10.1016/j.ebiom.2025.105978 · Cited by 5 · see citing works
Zhang, J., Yu, H., Shen, Y., Yang, X., & Wang, Y. (2024) · Molecules · doi:10.3390/molecules29215007 · Cited by 4 · see citing works
Shen, S., Yun, J., Song, C., Lu, Y., Shen, Z., & Li, F. (2025) · HORMONES · doi:10.1007/s42000-025-00656-0 · Cited by 1 · see citing works
Sailis, A. B., Noh, M. A. M., Leo, B. F., Faruqu, F. N., Yee, A. H. A., & Sim, M. S. (2026) · Molecular and Cellular Endocrinology · doi:10.1016/j.mce.2026.112786
Serres-Gómez, M., González‐Casaus, M. L., Olmedo, M., Fernández–Calle, P., Torrubiano, D. V., Iturzaeta, J. M., Almodóvar, B. G., & Soto, A. B. (2026) · Clinica Chimica Acta · doi:10.1016/j.cca.2026.121312
Endogenous androgens, physical and cognitive function in healthy midlife men: A systematic review
Keren, D., Hayek, R., Toledano, Y., Strauss, T., Springer, S., & Goshen, A. (2026) · Mechanisms of Ageing and Development · doi:10.1016/j.mad.2026.112257
- Authors
- Martínez-Escribano, A., Maroto-García, J., Ruiz-Galdón, M., Barrios-Rodríguez, R., Álvarez-Millán, J. J., Cabezas-Sánchez, P., Plaza-Andrades, I., Molina-Vega, M., Tinahones, F. J., Queipo-Ortuño, M. I., & Fernández-García, J. C.
- Affiliations
- Department of Surgery, Biochemistry and Immunology, Málaga University, 29010 Málaga, Spain
- Departmento de Medicina Preventiva y Salud Pública, Universidad de Granada, 18011 Granada, Spain
- Consulting Químico Sanitario (CQS Lab), 28521 Madrid, Spain
- Instituto de Investigación Biomédica de Málaga (ES)
- Department of Endocrinology and Nutrition, Virgen de la Victoria University Hospital, 29010 Malaga, Spain
- Instituto de Salud Carlos III (ES)
- Department of Medical Oncology, Virgen de la Victoria and Regional University Hospitals-IBIMA, UMA-CIMES, 29010 Málaga, Spain
- Corresponding author
- Tinahones, Francisco J.
- ORCID
- Venue
- Biomolecules
- Publisher
- MDPI AG
- Published
- 2020-12-19
- Volume
- 10
- Issue
- 12
- Pages
- 1697
- ISSN
- 2218-273X (ISSN-L)
- PMC
- PMC7765982
- OpenAlex
- W3112496624
- Cited by
- 11
- References
- 35
- Citations by year
- 2026: 3 · 2025: 3 · 2024: 1 · 2023: 1 · 2022: 1 · 2021: 2
- FWCI
- 0.75
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Biomolecules (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- LA Referencia (Red Federada de Repositorios Institucionales de Publicaciones Científicas) (publishedVersion, cc-by)
- RISalud Institutional Health Repository of Andalucía (submittedVersion, cc-by)
- Institutional Repository of the University of Granada (University of Granada) (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Servicio Andaluz de Salud (SAS) (B-0003-2017)
- Rio Hortega grant from Instituto de Salud Carlos III (CM18-00120)
- Miguel Servet Type II” program from ISCIII, co-funded by the Fondo Europeo de Desarrollo Regional – FEDER (CPI13/00003)
- Nicolas Monardes” research program of the Consejería de Salud, Junta de Andalucía, Spain (C-0030-2018)
- Servicio Andaluz de Salud (PI-0173-2013)
- Topics
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Hormonal and reproductive studies
- Health Sciences › Medicine › Physiology › Clinical Laboratory Practices and Quality Control
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
- Keywords
- serum testosterone, obesity, young men, immunoassay, liquid chromatography-tandem mass spectrometry, assay comparison, sex hormone-binding globulin, insulin resistance, diagnostic accuracy
- MeSH terms
- Adult, Chromatography, Chromatography, Liquid, Humans, Immunoassay, Male, Middle Aged, Obesity, Regression Analysis, Sensitivity and Specificity, Testosterone, Reproducibility of Results, Tandem Mass Spectrometry, Limit of Detection
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