title: Development and evaluation of a liquid chromatography tandem mass spectrometry method for simultaneous determination of salivary melatonin, cortisol and testosterone.
abstract: Circadian disruption can have several possible health consequences, but is not well studied. In order to measure circadian disruption, in relation to shift or night work, we developed a simple and sensitive method for the simultaneous determination of melatonin, cortisol and testosterone in human saliva. We used liquid-liquid extraction (LLE) followed by liquid chromatography coupled to electrospray tandem mass spectrometry (LC-ESI-MS/MS) recorded in positive ion mode. Saliva samples were collected by spitting directly into tubes and 250 μL were used for analysis. The limits of detection were 4.1 pmol/L, 0.27 nmol/L and 10.8 pmol/L for melatonin, cortisol, and testosterone, respectively. The developed method was sensitive enough to measure circadian rhythms of all 3 hormones in a pilot study among four healthy volunteers. It can therefor be used to study the impact of night work and working in artificial light on the workers circadian rhythms. To our knowledge this is the first LC-ESI-MS/MS method for simultaneous determination of salivary melatonin, cortisol and testosterone.
authors.affiliation: National Research Centre for the Working Environment, Lersø Parkallé 105, DK-2100 Copenhagen Ø, Denmark. maa@nrcwe.dk
authors.forename: Marie Aarrebo
authors.initials: MA
authors.lastname: Jensen
authors.forename: Ase Marie
authors.initials: AM
authors.lastname: Hansen
authors.forename: Peter
authors.initials: P
authors.lastname: Abrahamsson
authors.forename: Asger W
authors.initials: AW
authors.lastname: Nørgaard
mesh_terms: D000328:Adult; D002853:Chromatography, Liquid / Q000379:methods*; D002940:Circadian Rhythm; D004355:Drug Stability; D005260:Female; D006801:Humans; D006854:Hydrocortisone / Q000032:analysis*; D008297:Male; D008550:Melatonin / Q000032:analysis*; D008875:Middle Aged; D015203:Reproducibility of Results; D012463:Saliva / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013739:Testosterone / Q000032:analysis*
chemical_list: D013739:Testosterone; D008550:Melatonin; D006854:Hydrocortisone