title: Measurement of late-night salivary cortisol with an automated immunoassay system.
abstract: BACKGROUND
  Measurement of late-night salivary cortisol concentrations is increasingly used as a screening test in suspected Cushing's syndrome. Cortisol concentrations are typically extremely low in late-night samples and discordant assay-specific reference ranges have been reported. Therefore, the aim of our study was to assess the analytical performance of the first automated cortisol immunoassay specified for salivary measurements and to establish late-night sampling reference-range data for this test.
  
  
  METHODS
  Salivary cortisol was measured using the Roche Cobas Cortisol assay (Roche Diagnostics). Five salivary pools in different concentration ranges were used to assess the inter-assay imprecision of this test in a two-centre evaluation protocol including two reagent lots. Linearity was tested by serial dilution. Salivary samples were obtained at 23:00 h from 100 apparently healthy volunteers using a commercially available salivary sampling device (Salivette, Sarstedt). A subset of 20 samples was used for method comparison with isotope dilution liquid chromatography-tandem mass spectrometry.
  
  
  RESULTS
  Inter-assay coefficients of variation (n=20) between 11.6% and 40.4% were found for mean cortisol concentrations between 12.9 and 2.6 nmol/L, with an estimated functional sensitivity of approximately 5.0 nmol/L. The test also gave linear results in the lowest concentration range between 1.0 and 8.3 nmol/L. Mean late-night salivary cortisol of 5.0 nmol/L was found for healthy individuals; the absolute range was 1.4-16.7 nmol/L, and the 95th percentile was 8.9 nmol/L. Substantially lower concentrations were found with isotope dilution LC-MS/MS compared to immunoassay results (mean concentrations 1.8 and 4.4 nmol/L, respectively).
  
  
  CONCLUSIONS
  The automated assay investigated was found to offer acceptable analytical performance in the very low concentration range required for late-night salivary cortisol, despite a very short turn-around time. Using this assay, late-night salivary cortisol concentrations below 8.9 nmol/L are typically found in healthy volunteers.
authors.affiliation: Institute of Clinical Chemistry, Hospital of the University of Munich, Marchioninistrasse 15, 81377 Munich, Germany. michael.vogeser@med.uni-muenchen.de
authors.forename: Michael
authors.initials: M
authors.lastname: Vogeser
authors.forename: Jürgen
authors.initials: J
authors.lastname: Durner
authors.forename: Ewald
authors.initials: E
authors.lastname: Seliger
authors.forename: Christoph
authors.initials: C
authors.lastname: Auernhammer
mesh_terms: D000293:Adolescent; D000328:Adult; D015992:Body Mass Index; D003480:Cushing Syndrome / Q000175:diagnosis / Q000378:metabolism; D005260:Female; D006801:Humans; D006854:Hydrocortisone / Q000032:analysis*; D007118:Immunoassay / Q000379:methods; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D015203:Reproducibility of Results; D012463:Saliva / Q000737:chemistry*; D012680:Sensitivity and Specificity; D012737:Sex Factors; D013997:Time Factors
chemical_list: D006854:Hydrocortisone