Use of 3-(1,8-naphthalimido)propyl-modified silyl silica gel as a stationary phase for the high-performance liquid chromatographic separation of purine derivatives

Abstract

The use of a packing material, 3-(1,8-naphthalimido)propyl-modified silyl silica gel (NAIP), as a stationary phase for high-performance liquid chromatography, has been studied. NAIP behaved like a reversed-phase stationary phase with some π-π interaction. Purine derivatives, i.e., xanthine, hypoxanthine, uric acid, theobromine, theophylline and caffeine, were separated by a column packed with NAIP using an eluent of borate solution (pH 6.4)-MeOH (50:50, v/v). Of these, caffeine was selected as the target of the subsequent investigation and its determination was examined in commercially available medicinal drinks and pharmaceutical preparations. The average recoveries of caffeine were 98.0–107.4% for five drinks and 99.6–107.8% for five tablets and one powder. Subsequently, determination of caffeine and its metabolites in human plasma was examined. In twelve normal human plasma, caffeine levels ranged from 0.24 to 4.26 μ/ml. Time curves of plasma caffeine concentrations and those of its demethylated metabolite, 1,7-dimethylxanthine (1,7-DMX), after an oral ingestion of caffeine (200 mg) were measured by the proposed method and it was found that the maximum concentrations of caffeine and 1,7-DMX were obtained at 1-1.5 h and 3–6 h after ingestion, respectively.