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Measurement of caffeine and five of the major metabolites in urine by high-performance liquid chromatography/tandem mass spectrometry
Journal of mass spectrometry., 2005-03, Vol.40 (3), p.307-316
Weimann, Allan
Sabroe, Mads
Poulsen, Henrik E.
2005
Volltextzugriff (PDF)
Details
Autor(en) / Beteiligte
Weimann, Allan
Sabroe, Mads
Poulsen, Henrik E.
Titel
Measurement of caffeine and five of the major metabolites in urine by high-performance liquid chromatography/tandem mass spectrometry
Ist Teil von
Journal of mass spectrometry., 2005-03, Vol.40 (3), p.307-316
Ort / Verlag
Chichester, UK: John Wiley & Sons, Ltd
Erscheinungsjahr
2005
Quelle
MEDLINE
Beschreibungen/Notizen
Analysis of caffeine and its metabolites is of interest with respect to caffeine exposure, for kinetic and metabolism studies and for opportunistic in vivo estimation of drug metabolizing enzyme activity in humans and animals. For the latter, analysis is usually done by high‐performance liquid chromatography (HPLC) with UV detection. However, this method is close to the detection limit for certain of the metabolites and requires very long chromatography, 30–60 min. We have developed a fast method for the quantification of caffeine and its metabolites 1‐methylxanthine, 1‐methyluric acid, 1,7‐dimethyluric acid, 5‐acetylamino‐6‐amino‐3‐methyluracil (AAMU) and 5‐acetylamino‐6‐formylamino‐3‐methyluracil (AFMU) by HPLC tandem mass spectrometry (MS/MS) in urine that requires only its dilution with buffer and centrifugation before injection into the HPLC/MS/MS system. The chromatography lasts 7 min and is followed by 4.5 min for re‐equilibration of the HPLC column, giving a total analysis time of 11.5 min. The method provides a great sensitivity improvement with detection limits for all analytes ≤25 nM in real samples. Also, the analysis provides much improvement in capacity to ∼125 samples per 24 h. Intra‐ and inter‐day coefficients of variation of a single analysis are <6.5% for all the analytes. The inter‐day coefficient of variation of duplicate analyses is <4.8% for all analytes. The method is automated, including automated integration, and it is fast, robust and suitable for large‐scale investigations in humans and animals. Copyright © 2005 John Wiley & Sons, Ltd.
Sprache
Englisch
Identifikatoren
ISSN: 1076-5174
eISSN: 1096-9888
DOI: 10.1002/jms.785
Titel-ID: cdi_proquest_miscellaneous_67509338
Format
–
Schlagworte
Analysis
,
Biological and medical sciences
,
Caffeine
,
Caffeine - metabolism
,
Caffeine - urine
,
Chromatography, High Pressure Liquid
,
electrospray
,
General pharmacology
,
Humans
,
Medical sciences
,
metabolites
,
Molecular Structure
,
Pharmacology. Drug treatments
,
Reference Standards
,
Spectrometry, Mass, Electrospray Ionization
,
urine
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