Quantitation of hypoxanthine in plasma from patients with ischemic heart disease: adaption of a high-performance liquid chromatographic method.
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 100-5C18 column, with a mobile phase of KH2PO4 (300 mM, 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 microM, respectively).
Himmel
H M
HM
Pharmakologisches Institut, Universität-GHS Essen, Germany.
Sadony
V
V
Ravens
U
U
D000493:Allopurinol / Q000097:blood; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003327:Coronary Disease / Q000097:blood*; D006151:Guanosine / Q000097:blood; D006801:Humans; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000097:blood*; D007288:Inosine / Q000097:blood; D014529:Uridine / Q000097:blood; D019820:Xanthine; D014970:Xanthines / Q000097:blood
D007042:Hypoxanthines; D014970:Xanthines; D006151:Guanosine; D019820:Xanthine; D019271:Hypoxanthine; D007288:Inosine; D000493:Allopurinol; C064273:7-methylxanthine; D014529:Uridine