HPLC同时测定人尿中假尿苷次黄嘌呤含量

Determination on Concentration of Pseudouridine and Hypoxanthine in Human Serum by HPLC

目的 建立一种同时检测人尿中假尿苷、次黄嘌呤含量的高效液相色谱方法 (HPLC)。方法 色谱条件 :Spherigel-C1 8色谱柱 ;流动相 :0 0 2mol/LKH2 PO4缓冲液 ;流速 :1ml/min ;UV :2 5 4nm。结果 本法假尿苷线性范围为 1mg/L～ 1 6mg/L ,相关系数r =0 .9986 ,平均回收率 96 % ,最低检测浓度 0 2mg/L ;次黄嘌呤线性范围为1mg/L～ 2 0 /L ;相关系数r=0 .990 2 ;平均回收率 97% ;最低检测浓度为 0 3mg/L。结论 本法具有样品处理简单、稳定性高等特点 。

Objective To establish a method for the determination of concentration of Pseudouridine and hypoxanthine in the human urine(HPLC).Methods The concentration of Pseudouridine and hypoxanthine was assayed on a Spherigel-C 18 column with a mobile phase consisting of acetonitrile 0 02mol/L KH 2PO 4 at a flow rate of l ml/min and detected at UV 254nm.Results The calibration curve of Pseudouridine was linear over the range of l mg/L～16mg/L(r=0 9986),the average recovery of Pseudouridine was 96%,the limit of quantification was 0 2mg/L.The calibration curve of hypoxanthine was linear over the range of 1mg/L～20mg/L(r=0 9902),the average recovery of hypoxanthine was 97%,the limit of quantification was 0 3mg/L.Conclusion This method possessed the fectures of sample treatment simple and high stability.Clinical application demonstrated that this method had a better practicality.