Wu Jin, Fan Liuyin, Zhang Wei, Wang Qiuling, Cao Chengxi
Laboratory of Analytical Biochemistry & Bio-separation, School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
Se Pu. 2008 Sep;26(5):622-5.
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) x 20 s of pressure injection, 60.2 cm x 75 microm i.d. (50 cm effective length) of capillary, 214 nm of detection wavelength, 24 degrees C 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.
尿液的快速灵敏分析对于新生物标志物的发现和临床诊断具有明显的重要意义。在本研究中，描述了一种简单、快速且灵敏的移动反应边界（MRB）诱导堆积技术，用于通过毛细管区带电泳（CZE）分析尿液成分。MRB由酸性甘氨酸 - 盐酸作为样品缓冲液和碱性甘氨酸 - 氢氧化钠作为运行缓冲液形成。堆积和分离的最佳条件为：25 mmol/L甘氨酸 - 盐酸（pH 2.5）作为样品缓冲液，50 mmol/L甘氨酸 - 氢氧化钠（pH 12.3）作为运行缓冲液，施加电压15 kV，压力进样4.82 kPa（0.7 psi）×20 s，毛细管内径75 μm（有效长度50 cm）、长度60.2 cm，检测波长214 nm，分离温度24℃。在最佳条件下，MRB可显著提高尿液成分分析的灵敏度。观察到80多个堆积峰，与传统CZE相比，灵敏度提高了10倍以上，传统CZE只能观察到10个峰且灵敏度较差。结果表明，MRB诱导堆积是一种用于尿液成分分析的有用且有潜力的工具。此外，本方法易于操作，样品制备相当简单。
