RP-HPLC determination of four kinds of kidney disease markers in urine.docVIP

 PAGE \* MERGEFORMAT 16 RP-HPLC determination of four kinds of kidney disease markers in urine Author: Kong Yu, Zhao Xi, Wang Bo, Wu Hong, Ma state Keywords: Diabetic nephropathy Reversedphase high performance liquid chromatography assay for determining 4 kinds of markers of diabetic nephropathy in urine 【Abstract】 AIM: To establish a new reversedphase high performance liquid chromatography (RPHPLC) method for assay 4 kinds of nonproteinnitrogen markers of human diabetic nephropathy (DN), creatine (Cr), creatinine (Cn), urea (U) and uric acid (Ua) in DN patients urines. METHODS: The urine samples were assayed by RPHPLC on C18 column (200 mm × 4.6 mm, id 5 μm) 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 μmol / L, 10-200 μmol / L, 1-30 μmol / L, 10-1500 μmol / 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. 【Keywords】 reversed high performance liquid chromatography; diabetic nephropathy; nonprotein nitrogen; urine 【Abstract】 Objective: To establish diabetic patients with nephropathy urine of non-protein nitrogen metabolites (creatine, creatinine, urea and uric acid) by RP-HPLC separation and analysis. Methods: The Agilent C18 ( 200 mm × 4.6 mm, diameter 5 μm) column with mobile phase containing 30% (volume ratio) methanol and 10 mmol / L, pH 6.86 phosphate buffer solution, flow rate 0.9 mL / min, detection wavelength 200 nm, column temperature at room temperature. Results: In patients within 5 mi