应用高效毛细管电泳法测定2型糖尿病肾病患者尿微量白蛋白的含量

摘要

目的:应用毛细管电泳耦合二极管阵列检测器(CE-DAD)技术检测2型糖尿病肾病患者尿微量白蛋白(mALB)的含量.方法:实验过程中,对缓冲液的浓度、pH值、分离电压、最佳波长以及添加剂十二烷基硫酸钠(SDS)的浓度进行系统的研究,得到了最佳分离条件,即运行缓冲溶液为10 mmol/L硼酸盐+70 mmol/L SDS,pH=9.25,检测波长194 nm,分离电压15 kV,分离温度25℃的条件下,通过毛细管电泳压力进样8s,使空白对照组、糖尿病组和肾病组的mALB在15 min内均被完全分离.结果:糖尿病肾病组患者中mALB的含量均高于健康对照组,差异有显著统计学意义,并且随着患者病情的加重,患者体内的mALB的含量明显增高.结论:应用CE-DAD检测方法能够实现mALB良好分离,是一种可靠、高效的分离方法.%Objective:To establish a new method for determination of the microalbuminuria (mALB) in urine of patients with type 2 diabetic nephropathy (T2DN) by capillary electrophoresis (CE) coupled with diode array detector (DAD).Methods:Experimental parameters such as concentration and pH of buffer solution,the separation voltage,optimal wavelength and SDS concentration were systematically investigated.The optimum separation conditions were determined as:10 mmol/L borate buffer and 70 mmol/L SDS at pH 9.25 with 8 s hydrodynamic injection at 50 mbar,optimal wavelength 194 nm,applied voltage were set at 15 kV and working temperature at 25 ℃,and the mALB were completely separated within 15 min.Results:There was a significantly difference between the healthy control and the patients,and the concentration of mALB had significantly increased as the disease became worse.Conclusion:This CE-DAD method can achieve good accuracy and repetition for analyzing mALB.