Reversedª²phase high performance liquid chromatography assay for determining 4 kinds of markers of diabetic nephroª²pathy in urine

    KONG Yu£¬ZHAO Yongª²Xi, WANG Bo, WU Hong, MA Guoª²Ying

    Department of Bioengineer, Bioinformatics Research Center£¬ Key Laboratory of Ministry of Education for Biomedical Information Engineering, Xi¬ðan Jiaotong University, Xi¬ðan 710049, China, Department of Pharmaceutics, Fourth Military Medical University£¬Xi¬ðan 710033, China, Department of Applied Chemistry, Xi¬ðan University of Technology, Xi¬ðan 710048, China

    ¡¾Abstract¡¿ AIM: To establish a new reversedª²phase high performance liquid chromatography (RPª²HPLC) method for assay 4 kinds of nonª²proteinª²nitrogen markers of human diabetic nephroª²pathy (DN), creatine (Cr), creatinine (Cn), urea (U) and uric acid (Ua) in DN patients¬ð urines. METHODS: The urine samples were assayed by RPª²HPLC on C18 column (200 mm¡Á4.6 mm, id 5 ¦Ìm) at the ambient temperature and a wave length of 200 nm, using a 10 mmol/L pH 6.86 phosphate buffer [consisted 30%(V/V) methanol] as mobile phase with the flow rate of 0.9 mL/min. RESULTS: Baseline separation of the urine could be easily achieved in 5 min after a simple sample preparation. The method also displayed a high reproducibility (RSD values of retention time and peak area were no more than 3%) and the calibration curves showed good linearities in the range of 5-300 ¦Ìmol/L, 10-200 ¦Ìmol/L, 1-30 ¦Ìmol/L, 10-1500 ¦Ìmol/L for Cr, Cn, U and Ua respectively. CONCLUSION: This method has great potential for routine assay of clinical urine samples and monitoring DN early. ......