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An LC-MS/MS-based method for the simultaneous quantification of melatonin, cortisol and cortisone in saliva
Mirthe Lanfermeijer1, Lennart J van Winden1, Danielle E J Starreveld2
1Department of Laboratory Medicine, Netherlands Cancer Institute Amsterdam, the Netherlands.
Analytical Biochemistry
|March 2, 2024
Summary
Researchers developed a new method to measure melatonin, cortisol, and cortisone in saliva. This technique offers valuable biochemical insights into diurnal patterns, aiding in the study of conditions like depression and fatigue.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Endocrinology
Background:
- Diurnal pattern disturbances are linked to depression and fatigue.
- Salivary hormone analysis offers a non-invasive method for assessing diurnal patterns.
- Melatonin, cortisol, and cortisone are key hormones reflecting diurnal rhythms.
Purpose of the Study:
- To develop and validate a novel liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantifying salivary melatonin, cortisol, and cortisone.
- To establish a reliable and sensitive assay for assessing diurnal hormone profiles.
- To investigate potential interferences and assess the method's performance.
Main Methods:
- A liquid chromatography-tandem mass spectrometry (LC-MS/MS) approach was employed.
- Isotope dilution methodology and liquid-liquid extraction were used for sample preparation.
- Reversed-phase chromatography and multiple reaction monitoring (MRM) were utilized for quantification.
Main Results:
- The method achieved low limits of quantification (e.g., 0.010 nmol/L for melatonin) and detection.
- Imprecision was ≤14% for all measured hormones, with high correlation coefficients (≥0.964) in method comparison.
- Potential interference from prednisolone was identified, detectable via chromatogram review.
Conclusions:
- A validated LC-MS/MS method enables accurate and precise measurement of salivary melatonin, cortisol, and cortisone.
- This assay is suitable for studying diurnal hormone patterns in various populations, including cancer survivors.
- The method provides a valuable tool for research into conditions associated with circadian rhythm disruptions.
