CE with a new electrochemiluminescent detection system for separation and detection of proteins labeled with tris(1,10-phenanthroline) ruthenium(II)

✍ Scribed by Longhua Guo; Bin Qiu; Linlin Xue; Guonan Chen

• Book ID: 102191510
• Publisher: John Wiley and Sons
• Year: 2009
• Tongue: English
• Weight: 227 KB
• Volume: 30
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/elps.200800715

✦ Synopsis

Abstract

In this article, a CE with a new electrochemiluminescent (ECL) detection system was developed. A microfluidic ECL detection cell with less than 0.5 μL dead volumes was developed and used as detector for this system. A hydrofluoric acid‐etched porous joint was made at 8 mm from the outlet of the separation capillary to isolate the CE high voltage from ECL detection. The proposed CE‐ECL system was applied for separation and detection of some proteins labeled with tris(1,10‐phenanthroline) ruthenium(II). High efficiency ECL‐enhanced reagent, tripropylamine, was infused to the detection cell as coreactant by a micro‐infusion system to obtain maximum and stable ECL signal. The performance of this setup was illustrated by the analysis of tris(1,10‐phenanthroline) ruthenium(II)‐labeled proteins. The background electrolyte for protein detection was 20 mM Tris‐CH~3~COOH with 2.0% m/m PVP at pH 4.0. Under the optimal conditions, the corresponding LOD were 2.2×10^−10^ M for HSA, 4.4×10^−10^ M for casein (α‐S1) and 5.1×10^−10^ M for cytochrome c. The proposed method was also successfully used for the trace analysis of albumin in human urine without any pretreatment.