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

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 microL 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-CH3COOH with 2.0% m/m PVP at pH 4.0. Under the optimal conditions, the corresponding LOD were 2.2 x 10(-10) M for HSA, 4.4 x 10(-10) M for casein (alpha-S1) and 5.1 x 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.

Determination of enoxacin and ofloxacin by capillary electrophoresis with electrochemiluminescence detection in biofluids and drugs and its application to pharmacokinetics

“…Recently, CE-CL has been widely developed and become a useful tool for rapid detection of various analytes such as metal ions (Ren and Huang, 2001), amino acids , proteins (Zhi et al, 2007) and antigen-antibody complexes (Wang and Ren, 2005). The marriage of CE to ECL is a sensitive and effi cient analytical technique and has got excellent performance for the analysis of proteins Guo et al, 2009), amino acids (Li et al, 2006) and drugs (Zhao et al, 2004;Yuan et al, 2009;Liu et al, 2008aLiu et al, , 2009aXu et al, 2006;Deng et al, 2008).…”

Surface Enhanced Electrochemiluminescence of Ru(bpy)32+

“…Among all the ECL reagents, Ru(bpy) 3 2+ and its derivatives are the most representative and commonly used reagents due to their good water solubility, high electrochemical stability and the ability to be repeatedly regenerated in ECL reaction 19 20 . The Ru(bpy) 3 2+ -ECL reaction system has been widely used in the areas of immunoassay 21 22 23 , DNA quantification 24 25 26 , pharmaceutical study 25 27 and environmental detection 28 29 . Except for the wide use of the Ru(bpy) 3 2+ -ECL system in the scientific laboratories, the popularity of commercial ECL instruments, e.g.…”

Separation and determination of ephedra alkaloids in traditional Chinese medicine and human urines by capillary electrophoresis coupled with electrochemiluminescence detection

“…Comparing with the published results in urine applications, 2,6 this CE-ECL method can realize good separation and sensitive detection for E, ME, and PE in human urine. The recoveries at three different spiked concentration levels (20,200, and 400 mmol/L) were 101.5%-104.3% for E, 88.2%-101.3% for ME, and 97.6%-100.0% for PE, respectively. The RSDs of the peak area were less than 3.6%.…”