Minh Anh Thu Phan
Analysis of sex steroids in human tears using LC-MS and GC-MS: Considerations and developments to improve method sensitivity and accuracy
Phan, Minh Anh Thu; Gibson, Emma; Golebiowski, Blanka; Stapleton, Fiona; Jenner, Andrew M; Bucknall, Martin P
Authors
Emma Gibson Emma3.Gibson@uwe.ac.uk
Senior Lecturer in Optometry
Blanka Golebiowski
Fiona Stapleton
Andrew M Jenner
Martin P Bucknall
Abstract
Sex steroids play a role in regulation of tear film function and may exert their action locally at the ocular surface. However, measurement of sex steroids in tears is difficult due to small-volume tear samples and very low concentrations of the hormones. This short communication highlights what has been achieved to date in the analysis of tear sex steroids using ultra-performance LC-MS (UPLC-MS) as previously published, and reports further and more recent investigations toward optimising mass spectrometry method sensitivity and accuracy. The published UPLC-MS method successfully measured progesterone, androsterone glucuronide and 5α-androstane-3α,17β-diol in pooled basal tears of postmenopausal women, and fourteen sex steroid standards in methanol. Limitations included sub-optimal limits of detection (LOD) and lower limits of quantification (LLOQ) for some analytes (particularly oestrogens), exclusion of sample matrix effects and no use of internal standards. This update reports on further experiments carried out to improve sensitivity and accuracy. Sample matrix effects, internal standard spiking, and derivatisation with dansyl chloride and oximes were investigated. Dansylation significantly improved the LOD and LLOQ of oestrogens and their metabolites, by a factor of 10 for oestradiol and a factor of 5 for oestrone, but sensitivity of this updated method is not sufficient however for analysis of these oestrogens in human tears. Using gas chromatography-mass spectrometry (GC-MS) as an alternative technique to LC-MS, improved sensitivity for derivatised oestradiol is reported. This work demonstrates the need to develop higher sensitivity methods and points researchers towards specific MS ionisation techniques for future analysis of sex steroids in tears, in order to progress current understanding of the role of sex steroids in tear function and dry eye.
Citation
Phan, M. A. T., Gibson, E., Golebiowski, B., Stapleton, F., Jenner, A. M., & Bucknall, M. P. (2022). Analysis of sex steroids in human tears using LC-MS and GC-MS: Considerations and developments to improve method sensitivity and accuracy. Experimental Eye Research, 225, 109283. https://doi.org/10.1016/j.exer.2022.109283
Journal Article Type | Article |
---|---|
Acceptance Date | Oct 13, 2022 |
Online Publication Date | Oct 21, 2022 |
Publication Date | Dec 1, 2022 |
Deposit Date | Jan 19, 2023 |
Publicly Available Date | Oct 22, 2023 |
Journal | Experimental Eye Research |
Print ISSN | 0014-4835 |
Electronic ISSN | 1096-0007 |
Publisher | Elsevier |
Peer Reviewed | Peer Reviewed |
Volume | 225 |
Pages | 109283 |
DOI | https://doi.org/10.1016/j.exer.2022.109283 |
Keywords | Tear film, Sex steroids, GC-MS, Serum, LC-MS |
Public URL | https://uwe-repository.worktribe.com/output/10137565 |
Publisher URL | https://www.sciencedirect.com/science/article/pii/S0014483522003645?via%3Dihub |
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Analysis of sex steroids in human tears using LC-MS and GC-MS: Considerations and developments to improve method sensitivity and accuracy
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Copyright Statement
This is the author’s accepted manuscript of the article ‘Phan, M. A. T., Gibson, E., Golebiowski, B., Stapleton, F., Jenner, A. M., & Bucknall, M. P. (2022). Analysis of sex steroids in human tears using LC-MS and GC-MS: Considerations and developments to improve method sensitivity and accuracy. Experimental Eye Research, 225, 109283’. DOI: https://doi.org/10.1016/j.exer.2022.109283
The final published version is available here: https://www.sciencedirect.com/science/article/pii/S0014483522003645?via%3Dihub
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