毛细管电泳-激光诱导荧光检测测定脑啡肽方法的建立及其应用  

Development of capillary electrophoresis with laser-induced fluorescence detection for the determination of enkephalin and its application

作　　者：董标[1] 吴胜明[1] 梁月琴[1] 李瑞生[2] 董方霆[1] 

机构地区：[1]国家生物医学分析中心,北京100850 [2]解放军第302医院实验动物中心,北京100039

出　　处：《化学试剂》2008年第10期753-755,768,共4页Chemical Reagents

摘　　要：建立脑啡肽（ENK）柱前荧光素异硫氰酸盐（FITC）衍生化,毛细管区带电泳-激光诱导荧光检测（CZE-LIF）测定的方法,并对FITC衍生化的影响因素和毛细管电泳的分离条件进行优化。毛细管电泳的分离条件为：75μm（内径）×67 cm石英毛细管柱;60 mmol/L pH 9.4 Tris-硼砂缓冲液;电压350 kV/cm;柱温25℃;压力进样3 s;LIF检测,λex/λem为488/520nm。以脑啡肽（ENK）标准品的浓度和电泳峰面积建立标准工作曲线,得到回归方程A=0.494 66＋0.085 92c,线性范围是35.2-563.1 fmol/μL,（n=7）,r=0.998 5,检测限为1.6×10^-15mol/μL（S/N=3）。大鼠脑部微透析液样品中脑啡肽应用此方法测定含量为7.4×10^-15mol/μL。A method for determination of precolumn derivatization enkephalin with fluorescein isothiocyanate isomer I（FITC） by capillary zone electrophoresis with laser-induced fluorescence detection was developed.The derivatization affect factors and CZE separation conditions were investigated and optimized.Electrophoretic conditions were as follows：a fused-silica capillary,67 cm（60 cm to the detector）×75 μm i.d.,with a running buffer of 60 mm tris-sodium tetraborate at a pH of 9.4,and a temperature of 25 ℃;a 3-s pressure injection,with an applied voltage of 350 kV/cm,and the laser induced fluorescence detection was performed at an excitation wavelength of 488 nm,and an emission wavelength of 520 nm.A linear relationship between the concentration and peak area was obtained in the concentration range 35.2～563.1 f mol/μL with a correlation coefficient of 0.998 5,and the detection limit was 1.6×10^-15 mol/μL（S/N=3）.The applicability of the method to microdialysis sample was demonstrated.

关 键 词：毛细管电泳 激光诱导荧光检测 衍生化 脑啡肽 

分 类 号：O629.7[理学—有机化学]