尿中多胺的毛细管电泳激光诱导荧光测定法

Determination of polyamines in urine by capillary electrophoresis with laser induced fluorescence

目的 :建立高效毛细管区带电泳 (CZE)激光诱导荧光 (LIF)测定人尿中多胺 (POL)的新方法。方法 :样品用 5mmol/L的HClO4分离蛋白 ,样品中多胺与异硫氰酸荧光素 (FITC)完成衍生反应后 ,用四甲基乙二胺(TEMED)作内标 ,在 15mmol/L的硼酸盐缓冲液 (pH 8.5 )中 ,运用毛细管区带电泳进行色谱分离 ,以激光诱导荧光测定法测定多胺。结果 :多胺测定在 10～ 2 0 0 μg/L范围内具有良好的线性关系 (r =0 .996 ) ,最低检测限为 16μg/L ,日内变异系数 (CV) 3.5 1%～ 4 .6 1% ,日间变异系数 (CV) 3.74 %～ 4 .83% ,方法平均回收率为 96 .0 0 %～ 99.33%。结论 :本方法简便、快速、定量可靠 。

Objective To establish a new method to determine polyamines (POL) in urine using high performance capillary zone electrophoresis (CZE) with laser induced fluorescence. Methods Protein in the urine sample was separated by 5 mmol/L HClO 4, and then fluoresiso thiocyanate was added for the derivative reaction with POL in the sample. The N,N,N'N Tetramethylethylenediamine was added to the sample as the internal standard. The POL was separated by CZE and laser induced fluorescence in sodium tetraborate buffer (15 mmol/L,pH 8.5). Results The method had a good linearity in the POL range of 10～200 μg/L ( r =0.996); the minimum detectable concentration was 5 μg/L; the intra day coefficient of variation was 3.51%～4.61%;the inter day coefficient of variation was 3.74%(4.83%); and the average recovery rate was 96.00%～99.33%.Conclusion This method is simple, rapid and accurate,and may be used to the determine urine polyamines in clinical laboratories.