Simultaneous determination of melatonin and 6-hydroxymelatonin in human overnight urine by LC-MS/MS.
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1 septembre 2021
- doi: 10.1016/j.jchromb.2021.122938
- issn: 1570-0232
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info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jchromb.2021.122938
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info:eu-repo/semantics/altIdentifier/pmid/34521018
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info:eu-repo/semantics/altIdentifier/eissn/1873-376X
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info:eu-repo/semantics/altIdentifier/urn/urn:nbn:ch:serval-BIB_5CDFB8562D732
info:eu-repo/semantics/openAccess , CC BY-NC-ND 4.0 , https://creativecommons.org/licenses/by-nc-nd/4.0/
Mots-clés
CYP450; LC-MS/MS; Melatonin; PhenotypingSujets proches
Methoxyindolylethylacetamide AcetylmethoxytryptamineCiter ce document
G. Magliocco et al., « Simultaneous determination of melatonin and 6-hydroxymelatonin in human overnight urine by LC-MS/MS. », Serveur académique Lausannois, ID : 10.1016/j.jchromb.2021.122938
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Résumé
For the quantification of the pineal hormone melatonin and its metabolite, 6-hydroxymelatonin, in human overnight urine, a single accurate method by liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed. Urine samples were deconjugated using β-glucuronidase/arylsulfatase from Helix pomatia before solid phase extraction (SPE) purification. Chromatographic separation was performed using a reverse phase C18 column with a 7-minute gradient elution. Water was used as matrix to prepare the calibration standards, and deuterated analogues of melatonin and 6-hydroxymelatonin were used as internal standards. This newly developed method was validated in terms of linearity, accuracy, repeatability, intermediate precision, recovery, matrix effect, and stability according to the guidelines of the European Medicines Agency. The method was successfully applied to the analysis of overnight urine samples from 12 healthy volunteers, showing significant correlations of urinary melatonin and 6-hydroxymelatonin excretion rates with age. The urinary 6-hydroxymelatonin to melatonin ratio was also established and will be assessed in further studies as a potential endogenous metric of CYP1A2 activity.