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http://www.authorstream.com/Presentation/abikesh086-174281-capillary-electrophoresis-education-ppt-powerpoint/
Electro-chemiluminescence
• Excitation results from
electrochemical reaction
• Widely used in clinical analytes
• Ru(bpy)32+; emits photons-620nm
Ru(bpy)32+ + e- Ru(bpy)3+
Ru(bpy)32+ - e- Ru(bpy)33+
Ru(bpy)3+ + Ru(bpy)33+ Ru(bpy)32+* + Ru(bpy)32+
Ru(bpy)32+* Ru(bpy)32+ + hν
N - dealkylation
N - Alkylation
2+
Ru(bpy) 2+*
Ru(bpy) 3 3
hν λ = 620 nm
Experiment
• Solid phase extraction
• Electro-kinetic injection – HV 10 kV, 20 s, RT
• Uncoated Fused-Silica – 50cm, 50μm, 360μm
• Separation voltage 14 kV
• 15 mM running buffer
• MPI-A CE-ECL detector – 3-electrode system
– Working electrode; Pt wire (300-μm dia.)
– Reference electrode; Ag/AgCl (KCl saturated)
– Counter electrode; Pt wire
– PMT to capture photons (800 V)
Effect of Detection potential
Effect of running buffer pH
a = pH 7
b = pH 7.5
c = pH 8
d = pH 8.5
e = pH 9
Injection time - ECL intensity and Resolution
a = CIP
b = ENR
c = Rs
Linearity
Optimum conditions
Milk Samples
a = Blank Milk
b = Blank milk spiked; 15μg/kg CIP and 15μg/kg ENR;
Conclusions
• ECL detector is sensitive, simple and cheap
detection with a low background noise
• Detection limits of ENR (10 ng/ml) and CIP (15
ng/ml) were achieved
Acknowledgments
• Dr S.D Gilman
• CHEM 7750 Colleagues