Xiang Lei-Wen, Li Jing, Lin Jin-Ming, Li Hai-Fang
Beijing Key Laboratory of Microanalysis Methods and Instrumentation, Department of Chemistry, Tsinghua University, Beijing 100084, China.
Institute of Food and Fermentation Industries, Fuqing Branch of Fujian Normal University, Fuqing 350300, China.
J Pharm Anal. 2014 Apr;4(2):153-158. doi: 10.1016/j.jpha.2013.11.003. Epub 2013 Dec 12.
Creatinine, uric acid, hypoxanthine and xanthine are important diagnostic biomarkers in human urine for gouty arthritis or renal disease diacrisis. A simple method for simultaneous determination of these biomarkers in urine based on reversed-phase high-performance liquid chromatography (RP-HPLC) with ultraviolet (UV) detector was proposed. After pretreatment by dilution, centrifugation and filtration, the biomarkers in urine samples were separated by ODS-BP column by elution with methanol/50 mM NaHPO buffer solution at pH 5.26 (5:95). Good linearity between peak areas and concentrations of standards was obtained for the biomarkers with correlation coefficients in the range of 0.9957-0.9993. The proposed analytical method has satisfactory repeatability (the recovery of data in a range of creatinine, uric acid, hypoxanthine and xanthine was 93.49-97.90%, 95.38-96.45%, 112.46-115.78% and 90.82-97.13% with standard deviation of <5%, respectively) and the limits of detection (LODs, S/N≥3) for creatinine, uric acid, hypoxanthine, and xanthine were 0.010, 0.025, 0.050 and 0.025 mg/L, respectively. The established method was proved to be simple, accurate, sensitive and reliable for the quantitation of gouty arthritis' biomarkers in human urine samples. The ratio of creatinine to uric acid was found to be a possible factor for assessment of gouty arthritis.
肌酐、尿酸、次黄嘌呤和黄嘌呤是人体尿液中用于痛风性关节炎或肾脏疾病诊断的重要生物标志物。本文提出了一种基于反相高效液相色谱（RP-HPLC）结合紫外（UV）检测器同时测定尿液中这些生物标志物的简单方法。尿液样品经稀释、离心和过滤预处理后，采用ODS-BP柱，以pH 5.26的甲醇/50 mM NaHPO缓冲溶液（5:95）洗脱，分离尿液中的生物标志物。生物标志物的峰面积与标准品浓度之间具有良好的线性关系，相关系数在0.9957 - 0.9993范围内。所提出的分析方法具有令人满意的重复性（肌酐、尿酸、次黄嘌呤和黄嘌呤的数据回收率分别为93.49 - 97.90%、95.38 - 96.45%、112.46 - 115.78%和90.82 - 97.13%，标准偏差均<5%），肌酐、尿酸、次黄嘌呤和黄嘌呤的检测限（LODs，S/N≥3）分别为0.010、0.025、0.050和0.025 mg/L。该方法被证明对于定量检测人体尿液样品中痛风性关节炎的生物标志物简单、准确、灵敏且可靠。肌酐与尿酸的比值被发现是评估痛风性关节炎的一个可能因素。
