Document Detail

Quantitative determination of melatonin in human plasma and cerebrospinal fluid with high-performance liquid chromatography and fluorescence detection.
MedLine Citation:
PMID:  10960829     Owner:  NLM     Status:  MEDLINE    
A validated new and precise reversed-phase high-performance liquid chromatographic method for the determination of melatonin in human plasma and cerebrospinal fluid, with 5-fluorotryptamine as internal standard, is described. Liquid-liquid extraction with dichloromethane was performed under alkaline conditions. After evaporation of the organic solvent, the extract was dissolved in eluent and chromatographed on a base-deactivated octadecyl column, using an eluent composed of 650 mL potassium dihydrogenphosphate solution (0.07 mol/L water), adjusted to a pH of 3.0 with a 43% phosphoric acid solution, mixed with 350 mL methanol. Fluorescence detection at an excitation wavelength of 224 nm and an emission wavelength of 348 nm was used for quantitation. Melatonin and 5-fluorotryptamine chromatographed with retention times of 5.3 and 9. 3 min, respectively. Mean recoveries of 96% (n = 10) and 95% (n = 5) were found for melatonin in plasma and cerebrospinal fluid respectively. 5-Fluorotryptamine was found to have a mean recovery of 90% (n = 10) and 82% (n = 5) in plasma and cerebrospinal fluid, respectively. The repeatability coefficients of variation for both melatonin and 5-fluorotryptamine in plasma were 4-5% [five different samples (r = 5) on two consecutive days (n = 2)], with reproducibility coefficients of 1.6-7% (n = 2, r = 5) and 0.9-4% (n = 2, r = 5) for melatonin and internal standard, respectively. In cerebrospinal fluid the repeatability coefficient of variation of the extraction procedure was 5% (n = 1, r = 5) for melatonin and 7% (n = 1, r = 5) for 5-fluorotryptamine. The correlation coefficients of the calibration curves were 0.9998 (n = 2) in plasma at a concentration range of 0.108-25.9 ng/mL and 0.9994 (n = 2) at a concentration range of 0.108-25.9 ng/mL in cerebrospinal fluid. The limit of detection was determined at 8 pg/mL which enables to measure melatonin concentrations at physiological concentrations reached during daytime.
J Sastre Toraño; P v Rijn-Bikker; P Merkus; H J Guchelaar
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biomedical chromatography : BMC     Volume:  14     ISSN:  0269-3879     ISO Abbreviation:  Biomed. Chromatogr.     Publication Date:  2000 Aug 
Date Detail:
Created Date:  2000-09-25     Completed Date:  2000-09-25     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  8610241     Medline TA:  Biomed Chromatogr     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  306-10     Citation Subset:  IM    
Copyright Information:
Copyright 2000 John Wiley & Sons, Ltd.
Pharmaceutical and Toxicological Laboratory, Department of Pharmacy, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
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MeSH Terms
Chromatography, High Pressure Liquid / methods*
Melatonin / analysis*,  blood,  cerebrospinal fluid
Reference Standards
Reproducibility of Results
Sensitivity and Specificity
Spectrometry, Fluorescence
Tryptamines / analysis,  blood,  cerebrospinal fluid
Reg. No./Substance:
0/Tryptamines; 576-16-9/5-fluorotryptamine; 73-31-4/Melatonin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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