Safranow K, Machoy Z, Ciechanowski K
Department of Biochemistry and Chemistry, Pomeranian Academy of Medicine, Powstańców Wielkopolskich 72, Szczecin, PL-70-111, Poland.
Anal Biochem. 2000 Nov 15;286(2):224-30. doi: 10.1006/abio.2000.4790.
A high-pressure liquid chromatography method has been developed for the analysis in urinary calculi of six purines: uric acid, 2, 8-dihydroxyadenine, xanthine, hypoxanthine, allopurinol, and oxypurinol. Separation was conducted isocratically on a reversed-phase column, using 50 mM phosphate buffer (pH 5.5) / methanol (97/3, v/v) as mobile phase. Limits of detection, depending on compound, ranged from 7 to 28 microg/g stone weight. Hitherto, no reports have appeared on other purines present with uric acid in stones, due to lack of a sensitive and specific analytical method. We have now found that all calculi with more than 4% uric acid also contained 1-methyluric and 7-methyluric acids and trace amounts of hypoxanthine, xanthine, and 2,8-dihydroxyadenine. Accurate identification and quantitation of purines in urinary calculi are important for the diagnosis of rare metabolic diseases leading to urolithiasis (xanthinuria, dihydroxyadeninuria), as well as for prevention of iatrogenic complications during treatment with allopurinol of uric acid urolithiasis. The method may be used for reference purposes in clinical laboratories and for research on the pathogenesis of urolithiasis in disorders of purine metabolism.
已开发出一种高压液相色谱法，用于分析尿结石中的六种嘌呤：尿酸、2,8 - 二羟基腺嘌呤、黄嘌呤、次黄嘌呤、别嘌呤醇和氧嘌呤醇。在反相柱上进行等度分离，使用50 mM磷酸盐缓冲液（pH 5.5）/甲醇（97/3，v/v）作为流动相。检测限因化合物而异，范围为7至28微克/克结石重量。迄今为止，由于缺乏灵敏且特异的分析方法，尚无关于结石中与尿酸共存的其他嘌呤的报道。我们现已发现，所有尿酸含量超过4%的结石还含有1 - 甲基尿酸和7 - 甲基尿酸以及痕量的次黄嘌呤、黄嘌呤和2,8 - 二羟基腺嘌呤。准确鉴定和定量尿结石中的嘌呤对于诊断导致尿石症的罕见代谢疾病（黄嘌呤尿症、二羟基腺嘌呤尿症）以及预防尿酸尿石症患者使用别嘌呤醇治疗期间的医源性并发症很重要。该方法可用于临床实验室的参考目的以及嘌呤代谢紊乱中尿石症发病机制的研究。
