Li Mu-De, Cheng Tian-Lu, Tseng Wei-Lung
Department of Chemistry, National Sun Yat-Sen University, Kaohsiung, Taiwan.
Electrophoresis. 2009 Jan;30(2):388-95. doi: 10.1002/elps.200800364.
In this study, we describe the use of Tween 20-capped gold nanoparticles (AuNPs) as selective probes for the extraction of aminothiols from an aqueous solution. Tween 20 molecules noncovalently attached to the surface of AuNPs to form Tween 20-AuNPs were used for the selective extraction of aminothiols through the formation of Au-S bonds. After extraction and centrifugation, the aminothiols were detached from the surface of the AuNPs by adding DTT in a high concentration. We used this probe in combination with CE and UV absorption detection. On-line concentration and separation of the released aminothiols were performed by using 1.6% v/v poly(diallyldimethylammonium chloride) as an additive in CE. Under optimal extraction and stacking conditions, the LOD at a S/N of 3 were 28, 554, and 456 nM for glutathione (GSH), cysteine (Cys), and homocysteine (HCys), respectively. In comparison with the normal injection without the extraction procedure, approximately 2280-, 998-, and 904-fold improvements in the sensitivity were observed for GSH, Cys, and HCys, respectively. We have validated the application of our method on the basis of the analysis of GSH and HCys in human urine samples. It is believed that this approach has significant potential to be extended to clinical diagnosis.
在本研究中,我们描述了使用吐温20包覆的金纳米颗粒(AuNPs)作为从水溶液中萃取氨基硫醇的选择性探针。非共价附着在AuNPs表面形成吐温20-AuNPs的吐温20分子,通过形成Au-S键用于氨基硫醇的选择性萃取。萃取和离心后,通过加入高浓度的二硫苏糖醇(DTT)将氨基硫醇从AuNPs表面分离。我们将该探针与毛细管电泳(CE)和紫外吸收检测结合使用。在CE中使用1.6% v/v的聚二烯丙基二甲基氯化铵作为添加剂,对释放的氨基硫醇进行在线富集和分离。在最佳萃取和堆积条件下,谷胱甘肽(GSH)、半胱氨酸(Cys)和同型半胱氨酸(HCys)在信噪比为3时的检测限分别为28、554和456 nM。与未进行萃取步骤的常规进样相比,GSH、Cys和HCys的灵敏度分别提高了约2280倍、998倍和904倍。我们通过分析人尿液样本中的GSH和HCys验证了该方法的应用。据信,这种方法具有扩展到临床诊断的巨大潜力。