A Novel LC-MS/MS Method for the Quantification of Five Androgens in Saliva
Malcolm McTaggart, Joanne Adaway, James Hawley, Brian Keevil Clinical Biochemistry, Pathology, Wythenshawe Hospital, Manchester University NHS Foundation Trust, Manchester, UK
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Malcolm McTaggart (Presenter) Manchester University NHS Foundation Trust
Presenter Bio: I took up the role of Senior Clinical Scientist in Clinical Biochemistry at Wythenshawe Hospital, part of Manchester University NHS Foundation Trust in November 2018. I have previously worked in Clinical Biochemistry at East Kent Hospitals University NHS Foundation Trust and the Shrewsbury and Telford Hospital NHS Trust, since joining the National Health Service in 2008. Prior to that, I completed a PhD in Biochemistry and Molecular Cell Biology at the University of Dundee. I took up my current role as I was to keen to learn about and practice mass spectrometry and become involved in research and development work again.
Relevant Financial Disclosures
(within past 24 months)
No relevant financial relationship(s) to disclose.
Abstract
Background
Androgenic activity is mediated by a combination of classical and 11-oxygenated androgens. Saliva production is generally more convenient than blood collection for patients since it is less invasive and does not require the patient to visit hospital or primary care for phlebotomy. We have developed an LC-MS/MS assay for the simultaneous measurement of testosterone, androstenedione, 17-hydroxyprogesterone, 11-ketotestosterone and 11-hydroxyandrostenedione in saliva.
Methods
Samples (250 µL of unstimulated whole saliva) were prepared by supported liquid extraction with methyl tert-butyl ether (MTBE) and reconstituted in 40% methanol. Liquid chromatography was on a Waters T3 column using a water (0.1% formic acid and 2 mmol/L ammonium acetate) / acetonitrile gradient. A TQ-XS Waters mass spectrometer was used for quantification.
Results
Total run time was four minutes per sample. A full validation of the method was performed including precision analysis, assessment of the lower limit of quantitation, as well as linearity, matrix effect, recovery, carryover and interference studies. Additionally, results from this method were compared to a previously published LC-MS/MS method for the five salivary androgens using saliva samples from healthy volunteers.
Conclusion
We present a novel LC-MS/MS method for the quantification of five androgens in saliva.
= Discovery stage.
= Translation stage.
= Clinically available.
