Dermanowski · Chronobiology international 2022 · cross-sectional method validation study · n=21

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

Cited 17 times in the scientific literature.

Level 3 - non-randomized controlled study

Cross-sectional comparative diagnostic method validation in healthy volunteers

PubMed 35168448 · doi:10.1080/07420528.2021.2016796 · record verified 2026-08-30

What was done

Researchers developed and validated a liquid chromatography with tandem mass spectrometry (LC-MS/MS) method following European Medicines Agency (EMA) guidelines to measure melatonin in plasma and saliva. They recruited 21 male and female volunteers aged 26–54 years and collected matched plasma and saliva samples at five time points between 20:00 and 00:00 hours. Dim light melatonin onset (DLMO) was defined as the time melatonin concentrations exceeded 20 pg/mL in plasma and 7 pg/mL in saliva, and values between both matrices were compared.

What was found

Plasma and salivary melatonin concentrations were significantly correlated (r = 0.764, p < .001), with a plasma-to-saliva concentration ratio of 2.87. The mean DLMO timing was 21:30 ± 0:45 hours in plasma and 21:34 ± 1:00 hours in saliva. The correlation between DLMO timing derived from plasma and saliva profiles was r = 0.679 (p < .05).

Why it matters

Salivary measurement using LC-MS/MS offers a non-invasive, reliable alternative to blood sampling for determining DLMO, facilitating circadian phase assessment in clinical and research settings.

Limits

The sample size was small (n = 21) and restricted to healthy volunteers, lacking validation in clinical populations with circadian rhythm sleep-wake disorders. Sampling was limited to a 4-hour window (20:00 to 00:00), which may fail to capture onset in individuals with advanced or delayed sleep phase.

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