摘要
建立了测定CD45-FITC荧光抗体样品溶液中游离FITC浓度的毛细管电泳(CE)方法。熔融石英毛细管45 cm(有效长度30 cm)×75μm i.d.;0.05 mol·L-1磷酸氢二钠-磷酸二氢钠缓冲液(含5g·L-1PEG4000,p H=7.14);分离电压+12 k V;进样压力0.5 psi(3.45 k Pa),进样时间3.0 s;分离温度25℃;UV-Vis检测器检测波长200 nm。该方法能有效测定CD45-FITC荧光抗体样品溶液中游离的FITC含量,线性回归方程相关系数r=0.9900,检测限LOQ=0.20μg·m L-1。样品加标回收率为105.49%,相对标准偏差小于5.36%。该方法快速、灵敏、重现性好,能用于CD45-FITC荧光抗体样品溶液中游离FITC的快速测定。
A rapid capillary electrophoresis（ CE） method was developed for the determination of free fluorescein isothiocyanate（ FITC） in CD45-FITC fluorescent antibodysolution. The analyses were carried out on a CE system equipped with an UV-Vis detector and a PDA detector. Separation was performed in a bare fused-silica capillary of 45. 0 cm long（ 30. 0 cm to the detector,75μm i. d.） at 25℃,and a 0. 05 mol·L-1disodium hydrogen phosphate-sodium dihydrogen phosphate buffer solution（ containing 5g·L-1PEG4000,p H = 7. 14） was used as background electrolyte. The samples were introduced into the capillary under a pressure of 0. 5psi（ 3. 45 k Pa） for 3 s,and the separation voltage was set at ＋ 12 k V. The detection wavelength was fixed at 200 nm. In this approach,FITC could be determined in CD45-FITC fluorescent antibody solution. The correlation coefficient for linear regression equation（ r） for FITC standard sample was above 0. 9900. The detection limit was about 0. 20 μg·m L-1. The spiked recoveries were about105. 49%,and the relative standard deviation was kept within 5. 36%. The method is simple,sensitive,rapid,reliable and reproducible. It can be applied for the determination of FITC in CD45-FITC fluorescent antibodysolution.
出处
《化学研究与应用》
CAS
CSCD
北大核心
2015年第11期1666-1670,共5页
Chemical Research and Application
基金
安徽省高校省级优秀青年人才基金重点项目(2013SQRL051ZD)资助
安徽省自然科学基金项目(1308085QB24)资助
安徽省生化药物工程技术研究中心开放课题(BYEC1202
BYEC1301)资助
安徽省省级质量工程教学研究项目(2014jyxm211)资助
安徽省高等教育振兴计划项目(2014zytz014)资助
国家级大学生创新创业训练计划项目(201410367014)资助
蚌埠市社会科学规划项目(BB15B016)资助
