胶束电动毛细管电泳检测尿液转铁蛋白和白蛋白

摘要

Objective A simple,cheap and rapid method has been developed for the separation of transferrin(TRF)and albumin(Alb) by Micellar Eleetrokinetic Chromatography(MEKC).Methods Effects of separation conditions such as pH,running buffer type and concentration,surfactant types and concentration on the separation of TRF and Alb were investigated and optimized in detail.And MEKC was correlated with radioimmunoassay(RIA).Results Under the optimal conditions,the base lines separation of these proteins were achieved with satisfactory repeatability and high separation efficiency in 12 min using 50 mmol/L Tris buffer(pH 8.1) containing 10 mmol/L SDS,under 15 kV applied voltage.The linearity of the two proteins was in the range of 10-1000 mg/L.The detection limits in turn were 1.22 mg/L for TRF,1.05 mg/L for Alb with the RSD(n=6) of the migration time is<2.5% and the RSD(n=6) of peak area is lass than 5.4% for each proteins.And MEKC had better correlation with RIA,the correlative coefficient of TRF and Alb were 0.995 and 0.992,respectively.Conclusion MEKC separates TRF and Alb is a rapid,sensitive method with very small amount of samples and reagents needed.%目的 建立一种胶束电动毛细管电泳(Micellar Eleetrokinetic Chromatography,MEKC)快速检测尿液转铁蛋白(Transferrin,TRF)、白蛋白(Albumin,Alb)的方法.方法 详细考察并优化MEKC分离这两种蛋白质的影响因素,如缓冲溶液种类及浓度、pH、表面活性剂种类及浓度、分离电压、进样时间等.结果 当缓冲溶液为50 mmol/L三羟甲基氨基甲烷(Hydroxymethyl,Tris)(pH 8.1)、10 mmol/L 十二烷基硫酸钠(Sodium dodecyl sulfate,SDS)、分离电压为15 kV时,两种蛋白质在12 min内完成了很好的基线分离.在优化的条件下,检测TRF和Alb的线性范围为10～1 000 mg/L,检出限分别为1.22、1.05 mg/L,迁移时间重现性小于2.5%,面积的重现性小于5.4%.与放射免疫方法对比具有很好的相关性(TRF:r=0.995;Alb:r=0.992).结论 MEKC检测尿液TRF和Alb是一种快速、灵敏度高、试剂和样品用量少的方法.