Simultaneous measurement of allantoin, uric acid, xanthine and hypoxanthine in blood by high-performance liquid chromatography
Simultaneous measurement of allantoin, uric acid, xanthine and hypoxanthine in blood by high-performance liquid chromatography
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DOI：
10.1016/s0378-4347(97)00459-3
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发表时间：
1997-12-19
期刊：
影响因子：
--
通讯作者：
Kowalczyk, J
中科院分区：
文献类型：
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作者：
Czauderna, M;Kowalczyk, J
建立了一种测定反刍动物血浆中核酸和嘌呤分解代谢产物氧化嘌呤（即尿酸、黄嘌呤和次黄嘌呤）和尿囊素的高效液相色谱方法。血浆用10%三氯乙酸脱蛋白。该方法使测定氧化嘌呤没有衍生化。用2，4-二硝基苯肼转化为腙（GLX-DNPH）后测定尿囊素。采用两个反相C-18柱分离转化后的尿囊素、尿酸、黄嘌呤和次黄嘌呤衍生物。柱前衍生化和梯度洗脱与在205、254和360 nm处监测流出物的组合提供了用于研究血浆中的氧化嘌呤和尿囊素的简单且选择性的分析工具。HPLC分析的总运行时间为60 min。加入血浆中的嘌呤衍生物（即氧化嘌呤和尿囊素）的回收率为95 - 106%。当处理的样品在-10 ℃下储存7天时，嘌呤衍生物是稳定的。批内变异系数的低值（2.5-4.6%）和检测限的低值（0.187-0.004 nmol）表明该方法具有令人满意的精密度和灵敏度。(C)1997年Elsevier Science B.V.
A high-performance liquid chromatographic method for determining catabolism products of nucleic acids and purines, such as oxypurines (i.e. uric acid, xanthine and hypoxanthine) and allantoin in the blood plasma of ruminants was developed. The plasma was deproteinized with 10% trichloroacetic acid. The method enabled determination of oxypurines without derivatization. Allantoin was determined after conversion with 2,4-dinitrophenylhydrazine to a hydrazone (GLX-DNPH). Separation of converted allantoin, uric acid, xanthine and hypoxanthine derivatives was carried out using two reversed-phase C-18 columns. The combination of pre-column derivatization and gradient elution with monitoring of the effluent at 205, 254 and 360 nm provides a simple and selective analytical tool for studying oxypurines and allantoin in plasma. The total run time of the HPLC analysis was 60 min. The recovery of the purine derivatives (i.e. oxypurines and allantoin) added to the plasma was between 95 and 106%. Purine derivatives were stable when the processed samples were stored for 7 days at -10 degrees C. The low values of the intra-assay coefficient of variations (2.5-4.6%) and the low values of the detection Limits (0.187-0.004 nmol) point to the satisfactory precision and sensitivity of the method. (C) 1997 Elsevier Science B.V.