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Determination of salivary testosterone and androstendione by liquid chromatography–tandem mass spectrometry
- Source :
- Clinica Chimica Acta. 413:594-599
- Publication Year :
- 2012
- Publisher :
- Elsevier BV, 2012.
-
Abstract
- Background Salivary steroid measurements have the potential to provide a non-invasive assessment of serum free steroid concentrations. Salivary testing has been proposed as a possible substitute for serum testing partly because of easy sample collection. Methods We developed a liquid chromatography–tandem mass spectrometry (LC–MS/MS) assay for simultaneous measurement of testosterone and androstendione in saliva. We spiked 100 μL of saliva with isotope-labeled internal standards and performed extraction with diethyl ether. After evaporation both steroids were analyzed by LC–MS/MS operating in the positive mode electrospray ionization (ESI) after separation on a reversed-phase column. Results The calibration curves for analysis of testosterone and androstendione exhibited consistent linearity and reproducibility in the range of 20–1000 pmol/L. The limit of quantitation (LOQ) was 10 pmol/L for both steroids. The mean recovery of the analytes added to saliva ranged from 97 to 102%. Between-run CVs were 3.6–4.0% for testosterone and 4.8–8.0% for androstendione. The upper limit (the 97.5 percentile) of testosterone and androstendione reference ranges for males was 356 pmol/L and 344 pmol/L and that for females 39 pmol/L and 349 pmol/L, respectively. Conclusions This LC–MS/MS method has the required lower limit of quantification for analysis of salivary concentrations of testosterone and androstendione. The assay is rapid, reliable and simple to perform with a routine LC–MS/MS spectrometer with required sensitivity.
- Subjects :
- Male
Saliva
Analyte
Electrospray ionization
medicine.medical_treatment
Clinical Biochemistry
030209 endocrinology & metabolism
Mass spectrometry
01 natural sciences
Biochemistry
Steroid
03 medical and health sciences
0302 clinical medicine
Tandem Mass Spectrometry
Liquid chromatography–mass spectrometry
medicine
Humans
Testosterone
Detection limit
Chromatography
Chemistry
010401 analytical chemistry
Biochemistry (medical)
Androstenedione
General Medicine
0104 chemical sciences
Female
Sample collection
Chromatography, Liquid
Subjects
Details
- ISSN :
- 00098981
- Volume :
- 413
- Database :
- OpenAIRE
- Journal :
- Clinica Chimica Acta
- Accession number :
- edsair.doi.dedup.....83edd5ab077269fde18bc6b53709ce7c