Mass spectrometric quantification of urinary 6-sulfatoxymelatonin: age-dependent excretion and biological variation
M van Faassen, A van der Veen… - Clinical Chemistry and …, 2021 - degruyter.com
Clinical Chemistry and Laboratory Medicine (CCLM), 2021•degruyter.com
Objectives Regulators of circadian rhythm, including melatonin, influence fundamental
biological processes. Measuring the melatonin metabolite 6-sulfatoxymelatonin in urine can
estimate melatonin production. 6-sulfatoxymelatonin is mainly analyzed by immunoassays,
but these methods are hampered by cross-reactivity and poor reproducibility when used to
analyze small molecules. Therefore, we validated a high-throughput liquid chromatography
with tandem mass spectrometry (LC–MS/MS) method to quantify 6-sulfatoxymelatonin in …
biological processes. Measuring the melatonin metabolite 6-sulfatoxymelatonin in urine can
estimate melatonin production. 6-sulfatoxymelatonin is mainly analyzed by immunoassays,
but these methods are hampered by cross-reactivity and poor reproducibility when used to
analyze small molecules. Therefore, we validated a high-throughput liquid chromatography
with tandem mass spectrometry (LC–MS/MS) method to quantify 6-sulfatoxymelatonin in …
Objectives
Regulators of circadian rhythm, including melatonin, influence fundamental biological processes. Measuring the melatonin metabolite 6-sulfatoxymelatonin in urine can estimate melatonin production. 6-sulfatoxymelatonin is mainly analyzed by immunoassays, but these methods are hampered by cross-reactivity and poor reproducibility when used to analyze small molecules. Therefore, we validated a high-throughput liquid chromatography with tandem mass spectrometry (LC–MS/MS) method to quantify 6-sulfatoxymelatonin in urine. We evaluated age-dependent 24-h excretion of 6-sulfatoxymelatonin into urine and the biological variation of urinary excretion in healthy individuals.
Methods
The online solid phase extraction method combined with LC–MS/MS was validated according to international guidelines, and used to measure the excretion of 6-sulfatoxymelatonin into urine of 240 healthy individuals. Biological variation of 6-sulfatoxymelatonin excretion was examined in 10 healthy individuals.
Results
Urinary 6-sulfatoxymelatonin results were well within the validation criteria (interassay coefficient of variation: <5.4%, quantification limit: 0.2 nmol/L). There was an age-related decrease in 6-sulfatoxymelatonin excretion into 24-h urine [F(5, 234)=13.9; p<0.001]. Within-subject variation of 6-sulfatoxymelatonin was 39.2% in day urine, 15.1% in night urine, and 12.2% in 24-h urine. Between-subject variation was 39.1% in day urine, 37.9% in night urine, and 36.8% in 24-h urine.
Conclusions
This MS-based method enables straightforward, reproducible, and sensitive quantification of 6-sulfatoxymelatonin in urine. Urinary 6-sulfatoxymelatonin levels decreased with age. Biological variation of 6-sulfatoxymelatonin excretion into urine was high between subjects and lower within subjects, indicating that repeated measurements of 6-sulfatoxymelatonin in 24-h urine are needed in future studies.
De Gruyter
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