Czauderna M, Kowalczyk J
The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, Jabłonna.
J Chromatogr B Biomed Sci Appl. 1997 Dec 19;704(1-2):89-98. doi: 10.1016/s0378-4347(97)00459-3.
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 C18 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.
建立了一种高效液相色谱法，用于测定反刍动物血浆中核酸和嘌呤的分解代谢产物，如氧嘌呤（即尿酸、黄嘌呤和次黄嘌呤）和尿囊素。血浆用10%三氯乙酸脱蛋白。该方法能够在不进行衍生化的情况下测定氧嘌呤。尿囊素在与2,4-二硝基苯肼转化为腙（GLX-DNPH）后进行测定。使用两根反相C18柱对转化后的尿囊素、尿酸、黄嘌呤和次黄嘌呤衍生物进行分离。柱前衍生化与梯度洗脱相结合，并在205、254和360 nm处监测流出物，为研究血浆中的氧嘌呤和尿囊素提供了一种简单且具选择性的分析工具。HPLC分析的总运行时间为60分钟。添加到血浆中的嘌呤衍生物（即氧嘌呤和尿囊素）的回收率在95%至106%之间。当处理后的样品在-10℃下储存7天时，嘌呤衍生物稳定。测定内变异系数较低（2.5 - 4.6%）以及检测限较低（0.187 - 0.004 nmol）表明该方法具有令人满意的精密度和灵敏度。
