Assessment of dim light melatonin onset based on plasma and saliva samples

Melatonin (MELA) is a nocturnal hormone involved in the regulation of the circadian rhythm. MELA can be detected in plasma and saliva, and its salivary concentration strongly correlates with its plasma concentration. Dim light melatonin onset (DLMO) is considered to be the most accurate objective marker for assessing the circadian phase. The purpose of the study was to establish a method for the determination of MELA in plasma and saliva based on the liquid chromatography with tandem mass spectrometry (LC-MS/MS) and compare DLMO using both plasma and saliva matrices. The validation of the LC-MS/MS methods was performed in accordance with the European Medicines Agency (EMA) guideline. The study was conducted on a group of 21 volunteers, male and females, aged 26-54 years. Plasma and saliva were collected at five time points: between 20:00 and 00:00 hours. The MELA concentration was determined by the LC-MS/MS. The DLMO was considered as the point in time when MELA concentration exceeds 20 pg/mL in plasma and 7 pg/mL in saliva. The correlation coefficient between the plasma and salivary MELA concentration was r = 0.764 (p < .001). The ratio of the plasma/saliva MELA concentrations was 2.87. The mean time of the DLMO in the plasma was 21:30 +/- 0:45 hours, and in the saliva was as follows: 21:34 +/- 1:00 hours. The correlation between the DLMO, calculated based on the plasma and saliva MELA profiles, was r = 0.679 (p < .05). The determination of salivary MELA concentration using LC-MS/MS allows for the determination of the DLMO. Our method may be applied in clinical practice for the diagnosis and monitoring of circadian rhythm disorders.