CE with a new electrochemiluminescent detection system for separation and detection of proteins labeled with tris(1,10-phenanthroline) ruthenium(II).
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.
Guo
Longhua
L
Ministry of Education Key Laboratory of Analysis and Detection Technology for Food Safety Department of Chemistry, Fuzhou University, Fuzhou, P. R. China.
Qiu
Bin
B
Xue
Linlin
L
Chen
Guonan
G
D000419:Albuminuria / Q000652:urine; D004553:Electric Conductivity; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D016014:Linear Models; D008163:Luminescent Measurements / Q000379:methods*; D008297:Male; D009942:Organometallic Compounds / Q000737:chemistry*; D010618:Phenanthrolines / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry; D011506:Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012709:Serum Albumin / Q000032:analysis
D009942:Organometallic Compounds; D010618:Phenanthrolines; D010710:Phosphates; D011506:Proteins; D012709:Serum Albumin; C034282:tris-(1,10-phenanthroline)ruthenium; C018279:sodium phosphate