Abstract
A high-performance liquid chromatographic method is described for the separation and quantitation of several purine compounds, including hypoxanthine. The isocratic separation of a standard mixture of nine compounds is achieved within 20 min on a reversed-phase Nucleosil 100-5C 18 column, with a mobile phase of KH 2PO 4 (300 m M, pH 4.0)—methanol—acetonitrile—tetrahydrofuran (97.9:1:1:0.1, v/v). Uric acid, guanine, hypoxanthine, uridine, xanthine, allopurinol, inosine, guanosine and 7-methylxanthine were almost completely baseline-separated, with detection limits in the range 0.5–1.2 pmol per injection. The influence of the concentrations of buffer and tetrahydrofuran on the quality of separation are described. The within-day and the day-to-day precision were satisfactory ( e.g. coefficients of variation of less than 1.5 and ca. 6.0%, respectively, for peak heights). The recovery of [ 3H]hypoxanthine added to samples was 86 ± 1%. Hypoxanthine was quantified in human plasma samples obtained at various times during coronary artery bypass grafting. The hypoxanthine levels measured immediately after release of the aortic cross-clamp were significantly higher than those determined under control conditions (18.8 ± 7.0 and 3.4 ± 1.0 μ M, respectively).
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: Journal of Chromatography B: Biomedical Sciences and Applications
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
