Determination of melatonin in saliva using automated solid-phase extraction, high-performance liquid chromatography and fluorescence detection.
A sensitive bioanalytical method for the determination of melatonin in saliva by solid-phase extraction (SPE), high-performance liquid chromatography (HPLC) and fluorescence detection has been developed and validated. Saliva was collected with a Salivette sampling device (Sarstedt) and a mixed-mode SPE column was used for the extraction of melatonin and internal standard (N-acetyl-6-methoxytryptamine) from the saliva. Chromatographic separation was performed using a HyPurity C18 LC column (150 x 2.1 mm) with mobile phase acetonitrile-ammonium hydrogen carbonate buffer, 0.015 M, pH 6.8 (23:77, v/v). Excitation and emission wavelengths were set to 285 nm and 345 nm, respectively. The within-day precision for the method at 50 pmol/L was 7.9 % and the between-day precision was 10.5 %. The limit of quantification was 50 pmol/L.
Römsing
S
S
Department of Clinical Chemistry, Falun Central Hospital, Falun, Sweden. srm@du.se
Bökman
F
F
Bergqvist
Y
Y
D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D006801:Humans; D008550:Melatonin / Q000008:administration & dosage / Q000032:analysis* / Q000592:standards; D012015:Reference Standards; D012463:Saliva / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence
D008550:Melatonin