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An improved method for measurement of testosterone in human plasma and saliva by ultra-performance liquid chromatography-tandem mass spectrometry
The aim of the study was to develop and validate a practical assay of clinically relevant testosterone levels in human plasma and saliva. We performed ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis on Atlantis dC18 steel column using a mobile phase of 2-mM amm...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Wolters Kluwer - Medknow
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7305780/ https://www.ncbi.nlm.nih.gov/pubmed/32587818 http://dx.doi.org/10.4103/japtr.JAPTR_162_19 |
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author | Alvi, Syed N. Hammami, Muhammad M. |
author_facet | Alvi, Syed N. Hammami, Muhammad M. |
author_sort | Alvi, Syed N. |
collection | PubMed |
description | The aim of the study was to develop and validate a practical assay of clinically relevant testosterone levels in human plasma and saliva. We performed ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis on Atlantis dC18 steel column using a mobile phase of 2-mM ammonium acetate and acetonitrile (20:80, v: v) that was delivered at 0.3 ml/min. After adding d3-testosterone as an internal standard (IS), we extracted plasma and salivary samples with methyl tert-butyl ether. Mass spectrometry was performed in electrospray positive-ion mode. Targeted ion transitions were examined at m/z 289.18 → 97.04 and 292.24 → 97.04 for testosterone and IS, respectively. We validated the method according to the US Food and Drug Administration guidelines. Elution times for testosterone and IS were both around 1.35 min. Testosterone level was linearly associated (r(2) = 0.9975 and 0.9958) with peak area ratio of testosterone to IS between 0.5–50 ng/ml and 10–400 pg/ml in plasma and saliva, respectively. The coefficient of variation and bias were ≤12.6% and ≤±12.1% in plasma and ≤10.2% and ≤±5.3% in saliva. The extraction recovery of testosterone was ≥92% from plasma and ≥94% from saliva. Testosterone was stable (≥91%) for 24 h at room temperature and for 8 weeks at −20°C in both plasma and salivary samples. We report a simple, validated, UPLC-MS/MS assay that can be used to determine clinically relevant levels of testosterone in human plasma and saliva. |
format | Online Article Text |
id | pubmed-7305780 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Wolters Kluwer - Medknow |
record_format | MEDLINE/PubMed |
spelling | pubmed-73057802020-06-24 An improved method for measurement of testosterone in human plasma and saliva by ultra-performance liquid chromatography-tandem mass spectrometry Alvi, Syed N. Hammami, Muhammad M. J Adv Pharm Technol Res Original Article The aim of the study was to develop and validate a practical assay of clinically relevant testosterone levels in human plasma and saliva. We performed ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis on Atlantis dC18 steel column using a mobile phase of 2-mM ammonium acetate and acetonitrile (20:80, v: v) that was delivered at 0.3 ml/min. After adding d3-testosterone as an internal standard (IS), we extracted plasma and salivary samples with methyl tert-butyl ether. Mass spectrometry was performed in electrospray positive-ion mode. Targeted ion transitions were examined at m/z 289.18 → 97.04 and 292.24 → 97.04 for testosterone and IS, respectively. We validated the method according to the US Food and Drug Administration guidelines. Elution times for testosterone and IS were both around 1.35 min. Testosterone level was linearly associated (r(2) = 0.9975 and 0.9958) with peak area ratio of testosterone to IS between 0.5–50 ng/ml and 10–400 pg/ml in plasma and saliva, respectively. The coefficient of variation and bias were ≤12.6% and ≤±12.1% in plasma and ≤10.2% and ≤±5.3% in saliva. The extraction recovery of testosterone was ≥92% from plasma and ≥94% from saliva. Testosterone was stable (≥91%) for 24 h at room temperature and for 8 weeks at −20°C in both plasma and salivary samples. We report a simple, validated, UPLC-MS/MS assay that can be used to determine clinically relevant levels of testosterone in human plasma and saliva. Wolters Kluwer - Medknow 2020 2020-04-22 /pmc/articles/PMC7305780/ /pubmed/32587818 http://dx.doi.org/10.4103/japtr.JAPTR_162_19 Text en Copyright: © 2020 Journal of Advanced Pharmaceutical Technology & Research http://creativecommons.org/licenses/by-nc-sa/4.0 This is an open access journal, and articles are distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as appropriate credit is given and the new creations are licensed under the identical terms. |
spellingShingle | Original Article Alvi, Syed N. Hammami, Muhammad M. An improved method for measurement of testosterone in human plasma and saliva by ultra-performance liquid chromatography-tandem mass spectrometry |
title | An improved method for measurement of testosterone in human plasma and saliva by ultra-performance liquid chromatography-tandem mass spectrometry |
title_full | An improved method for measurement of testosterone in human plasma and saliva by ultra-performance liquid chromatography-tandem mass spectrometry |
title_fullStr | An improved method for measurement of testosterone in human plasma and saliva by ultra-performance liquid chromatography-tandem mass spectrometry |
title_full_unstemmed | An improved method for measurement of testosterone in human plasma and saliva by ultra-performance liquid chromatography-tandem mass spectrometry |
title_short | An improved method for measurement of testosterone in human plasma and saliva by ultra-performance liquid chromatography-tandem mass spectrometry |
title_sort | improved method for measurement of testosterone in human plasma and saliva by ultra-performance liquid chromatography-tandem mass spectrometry |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7305780/ https://www.ncbi.nlm.nih.gov/pubmed/32587818 http://dx.doi.org/10.4103/japtr.JAPTR_162_19 |
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