Circadian Biomarkers in Humans: Methodological Insights into the Detection of Melatonin and Cortisol

Circadian rhythms are intrinsic, with roughly 24 h oscillations that coordinate many physiological functions and are increasingly recognized as key determinants of human health. When these rhythms become misaligned, there is an increased risk for neurodegenerative and psychiatric disorders, metaboli...

Full description

Saved in:
Bibliographic Details
Main Authors: Cene Skubic, Urša Zevnik, Katarina Nahtigal, Leja Dolenc Grošelj, Damjana Rozman
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Biomolecules
Subjects:
Online Access:https://www.mdpi.com/2218-273X/15/7/1006
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849733268121845760
author Cene Skubic
Urša Zevnik
Katarina Nahtigal
Leja Dolenc Grošelj
Damjana Rozman
author_facet Cene Skubic
Urša Zevnik
Katarina Nahtigal
Leja Dolenc Grošelj
Damjana Rozman
author_sort Cene Skubic
collection DOAJ
description Circadian rhythms are intrinsic, with roughly 24 h oscillations that coordinate many physiological functions and are increasingly recognized as key determinants of human health. When these rhythms become misaligned, there is an increased risk for neurodegenerative and psychiatric disorders, metabolic syndrome, sleep disturbances, and even certain cancers. The hormones, melatonin that rises in the evening and cortisol that peaks shortly after awakening, represent crucial biochemical markers of the circadian phase. This review systematically evaluates contemporary techniques for quantifying melatonin and cortisol, comparing biological matrices (blood, saliva, urine) alongside analytical platforms. Special focus is placed on two clinically informative markers: Dim Light Melatonin Onset (DLMO) and the Cortisol Awakening Response (CAR). We compared immunoassays with liquid chromatography tandem mass spectrometry (LC MS/MS), highlighting differences in sensitivity, specificity, and laboratory feasibility. Potential confounders, including ambient light, body posture, and exact sampling times—are discussed in detail, to show the capacity of providing the most reliable results. By emphasizing the need for standardized protocols and controlled sampling conditions, this review provides essential guidance for researchers and clinicians aiming to assess the circadian biomarkers melatonin and cortisol with precision since they can be used in clinical practice as diagnostic and prognostic tools for assessing numerous pathologies.
format Article
id doaj-art-5f45ba8c2bb64306bf842684e1985c2d
institution DOAJ
issn 2218-273X
language English
publishDate 2025-07-01
publisher MDPI AG
record_format Article
series Biomolecules
spelling doaj-art-5f45ba8c2bb64306bf842684e1985c2d2025-08-20T03:08:05ZengMDPI AGBiomolecules2218-273X2025-07-01157100610.3390/biom15071006Circadian Biomarkers in Humans: Methodological Insights into the Detection of Melatonin and CortisolCene Skubic0Urša Zevnik1Katarina Nahtigal2Leja Dolenc Grošelj3Damjana Rozman4Center for Functional Genomics and Bio-Chips, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, Zaloška cesta 4, 1000 Ljubljana, SloveniaCenter for Functional Genomics and Bio-Chips, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, Zaloška cesta 4, 1000 Ljubljana, SloveniaCenter for Functional Genomics and Bio-Chips, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, Zaloška cesta 4, 1000 Ljubljana, SloveniaInstitute of Clinical Neurophysiology, University Medical Centre Ljubljana, 1000 Ljubljana, SloveniaCenter for Functional Genomics and Bio-Chips, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, Zaloška cesta 4, 1000 Ljubljana, SloveniaCircadian rhythms are intrinsic, with roughly 24 h oscillations that coordinate many physiological functions and are increasingly recognized as key determinants of human health. When these rhythms become misaligned, there is an increased risk for neurodegenerative and psychiatric disorders, metabolic syndrome, sleep disturbances, and even certain cancers. The hormones, melatonin that rises in the evening and cortisol that peaks shortly after awakening, represent crucial biochemical markers of the circadian phase. This review systematically evaluates contemporary techniques for quantifying melatonin and cortisol, comparing biological matrices (blood, saliva, urine) alongside analytical platforms. Special focus is placed on two clinically informative markers: Dim Light Melatonin Onset (DLMO) and the Cortisol Awakening Response (CAR). We compared immunoassays with liquid chromatography tandem mass spectrometry (LC MS/MS), highlighting differences in sensitivity, specificity, and laboratory feasibility. Potential confounders, including ambient light, body posture, and exact sampling times—are discussed in detail, to show the capacity of providing the most reliable results. By emphasizing the need for standardized protocols and controlled sampling conditions, this review provides essential guidance for researchers and clinicians aiming to assess the circadian biomarkers melatonin and cortisol with precision since they can be used in clinical practice as diagnostic and prognostic tools for assessing numerous pathologies.https://www.mdpi.com/2218-273X/15/7/1006melatoninDLMOcortisolLC–MS/MSELISAcircadian rhythms
spellingShingle Cene Skubic
Urša Zevnik
Katarina Nahtigal
Leja Dolenc Grošelj
Damjana Rozman
Circadian Biomarkers in Humans: Methodological Insights into the Detection of Melatonin and Cortisol
Biomolecules
melatonin
DLMO
cortisol
LC–MS/MS
ELISA
circadian rhythms
title Circadian Biomarkers in Humans: Methodological Insights into the Detection of Melatonin and Cortisol
title_full Circadian Biomarkers in Humans: Methodological Insights into the Detection of Melatonin and Cortisol
title_fullStr Circadian Biomarkers in Humans: Methodological Insights into the Detection of Melatonin and Cortisol
title_full_unstemmed Circadian Biomarkers in Humans: Methodological Insights into the Detection of Melatonin and Cortisol
title_short Circadian Biomarkers in Humans: Methodological Insights into the Detection of Melatonin and Cortisol
title_sort circadian biomarkers in humans methodological insights into the detection of melatonin and cortisol
topic melatonin
DLMO
cortisol
LC–MS/MS
ELISA
circadian rhythms
url https://www.mdpi.com/2218-273X/15/7/1006
work_keys_str_mv AT ceneskubic circadianbiomarkersinhumansmethodologicalinsightsintothedetectionofmelatoninandcortisol
AT ursazevnik circadianbiomarkersinhumansmethodologicalinsightsintothedetectionofmelatoninandcortisol
AT katarinanahtigal circadianbiomarkersinhumansmethodologicalinsightsintothedetectionofmelatoninandcortisol
AT lejadolencgroselj circadianbiomarkersinhumansmethodologicalinsightsintothedetectionofmelatoninandcortisol
AT damjanarozman circadianbiomarkersinhumansmethodologicalinsightsintothedetectionofmelatoninandcortisol