Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Prediction of Human Serum Concentration-Time Profiles of Golimumab and Ustekinumab Using Pharmacokinetic Data From Common Marmosets With Assessment of Anti-Drug Antibodies.

Biopharmaceutics & drug disposition·2026
Same author

The landscape of taste receptors in glioblastoma.

Biochemical pharmacology·2026
Same author

Endogenous melatonin partial metabolic clearance as a potential endogenous marker of CYP1A2 activity.

British journal of clinical pharmacology·2026
Same author

Translational applicability of human blood-brain barrier spheroid models for the development of brain-penetrant therapeutic antibodies.

Fluids and barriers of the CNS·2026
Same author

Contrasting cannabinoid receptor 2 (CB2R)-mediated responses in two different models of Blood Brain Barrier in the context of HIV.

Research square·2026
Same author

Brain-derived neurotrophic factor supports pericyte and vascular homeostasis in the aging brain.

Acta neuropathologica communications·2025

Related Experiment Video

Updated: Sep 25, 2025

Integration of Miniaturized Solid Phase Extraction and LC-MS/MS Detection of 3-Nitrotyrosine in Human Urine for Clinical Applications
08:41

Integration of Miniaturized Solid Phase Extraction and LC-MS/MS Detection of 3-Nitrotyrosine in Human Urine for Clinical Applications

Published on: July 14, 2017

9.5K

Development and Validation of an LC-MS/MS-Based Method for Quantifying Urinary Endogenous 6-Hydroxymelatonin.

Seiya Ohki1, Miyu Kunimatsu1, Shingo Ogawa1

  • 1Laboratory of Clinical Pharmacy and Experimental Therapeutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences.

Chemical & Pharmaceutical Bulletin
|May 1, 2022
PubMed
Summary

Accurately measuring melatonin (MEL) secretion requires quantifying urinary metabolites. This study developed an optimized LC-MS/MS method for 6-hydroxymelatonin (6-O-MEL) and its conjugates, improving circadian rhythm disorder diagnostics.

Keywords:
6-hydroxymelatoninLC–tandem mass spectrometrydeconjugationmelatoninurine

More Related Videos

Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules
07:35

Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules

Published on: December 16, 2021

2.4K
High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry
10:17

High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry

Published on: April 23, 2019

9.8K

Related Experiment Videos

Last Updated: Sep 25, 2025

Integration of Miniaturized Solid Phase Extraction and LC-MS/MS Detection of 3-Nitrotyrosine in Human Urine for Clinical Applications
08:41

Integration of Miniaturized Solid Phase Extraction and LC-MS/MS Detection of 3-Nitrotyrosine in Human Urine for Clinical Applications

Published on: July 14, 2017

9.5K
Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules
07:35

Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules

Published on: December 16, 2021

2.4K
High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry
10:17

High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry

Published on: April 23, 2019

9.8K

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Chronobiology

Background:

  • Endogenous melatonin (MEL) secretion is crucial for managing circadian rhythm sleep disorders.
  • Urinary MEL metabolites, primarily 6-hydroxymelatonin (6-O-MEL) sulfate (S-O-MEL) and 6-O-MEL glucuronate, reflect MEL secretion levels.
  • Accurate quantification requires enzymatic deconjugation and sensitive detection methods like LC-tandem mass spectrometry (LC-MS/MS).

Purpose of the Study:

  • To develop and optimize an LC-MS/MS-based quantification method for 6-O-MEL in human urine.
  • To establish optimal enzymatic deconjugation conditions for urinary MEL metabolites.
  • To provide a reliable method for assessing total MEL secretion levels.

Main Methods:

  • Optimization of LC-MS/MS conditions for 6-O-MEL measurement.
  • Determination of optimal enzymatic deconjugation conditions using Roche Glucuronidase/Arylsulfatase.
  • Validation of the method using human urine samples and analysis of 24-h urine collections.

Main Results:

  • Optimal deconjugation of S-O-MEL achieved at 37°C, pH 4.0, with 60 min reaction time, yielding 102.1% recovery.
  • Established minimum enzyme units (5944 units) for human urine samples.
  • Demonstrated high accuracy (-3.60--0.47% relative error) and precision (<6.80% standard deviation) for 6-O-MEL determination.
  • Measured total urinary MEL metabolite excretion at 6.70-11.28 µg in three subjects.

Conclusions:

  • An efficient LC-MS/MS method for quantifying total 6-O-MEL in human urine was successfully developed.
  • The optimized deconjugation and LC-MS/MS conditions ensure accurate and precise measurement of MEL metabolites.
  • This method facilitates improved assessment of MEL secretion for diagnosing and treating circadian rhythm disorders.