应用移动反应界面富集技术进行毛细管电泳尿液指纹分析

Urine profiling by capillary electrophoresis with online stacking of moving reaction boundary

快速灵敏的尿液指纹图谱分析对于临床诊断中发现新的生物标记至关重要。该文建立了一种简便、快速、灵敏的移动反应界面(MRB)介导的富集技术进行毛细管电泳尿液指纹图谱分析。MRB由25mmol/L甘氨酸(Gly)-HCl(pH2.5)作为样品缓冲液和50mmol/LGly-NaOH(pH12.3)作为电泳缓冲液形成。与常规的毛细管区带电泳只能观察到尿液中不到10个峰相比,采用MRB可以观察到超过80个峰并将检测灵敏度提高了至少十几倍,显示该方法对于代谢组学分析具有重要的意义。

Rapid and sensitive profiling of urine holds evident significance for the discovery of new biomarkers and clinical diagnosis.In this study,a simple,rapid and sensitive moving reaction boundary（MRB）-induced stacking technique is described for the analysis of urine profiling by capillary zone electrophoresis（CZE）.The MRB was formed with acidic Gly-HCl as the sample buffer and alkaline Gly-NaOH as the running buffer.The optimum conditions of stacking and separation were：25 mmol/L Gly-HCl（pH 2.5）as the sample buffer and 50 mmol/L Gly-NaOH（pH 12.3）as the running buffer,15 kV of applied voltage,4.82 kPa（0.7 psi）×20 s of pressure injection,60.2 cm×75 μm i.d.（50 cm effective length）of capillary,214 nm of detection wavelength,24 ℃of separation temperature.Under the optimum conditions,the MRB can significantly improve the sensitivity of urine profiling.More than 80 stacking peaks have been observed and more than 10 fold sensitivity enhancement was achieved in contrast with the conventional CZE,which with only ten observable peaks and poor sensitivity.The results indicate that the MRB-induced stacking is a useful and potential tool for urine profiling.Furthermore,the present method is easy to perform,and the preparation of sample is quite simple.