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  • Title: Simultaneous determination of caffeine and its primary demethylated metabolites in human plasma by high-performance liquid chromatography.
    Author: Tanaka E.
    Journal: J Chromatogr; 1992 Mar 27; 575(2):311-4. PubMed ID: 1629311.
    Abstract:
    An improved high-performance liquid chromatographic method for the simultaneous determination of caffeine and its three primary metabolites (theophylline, theobromine and paraxanthine) in human plasma is described. The four substances were separated on a reversed-phase column (5 microns TSK gel ODS-80TM, 150 mm x 4.6 mm I.D.) by use of the mobile phase methanol-0.1 M NaH2PO4 (30:70, v/v) with a flow-rate of 0.8 ml/min. Absorbance was monitored at 274 nm. The detection limit was 5 ng/ml for theobromine and caffeine and 10 ng/ml for paraxanthine and theophylline. The linearity and reproducibility were sufficient for drug monitoring of caffeine and its primary methylxanthines.
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