Determination of ascorbic acid in individual rat hepatocyte cells based on capillary electrophoresis with electrochemiluminescence detection.
A novel method to detect ascorbic acid (AA) in individual rat hepatocyte cells was developed by combining CE with electrochemiluminescence (ECL) based on tris(2,2'-bipyridine) ruthenium(II) (Ru(bpy)(3)2+). A single cell, followed by 0.1% SDS as cell lysis solution, was injected into the inlet of the separation capillary by electromigration. After optimizing the analytical conditions, the RSDs of migration time and peak height were 0.38% and 2.6% for 1.0x10(-5) M AA (n=10), respectively. The linear range of AA was from 1.0x10(-8) to 5.0x10(-5) M with a correlation coefficient of 0.9979 and the LOD (S/N=3) was estimated to be 1.0x10(-8) M. This method has been successfully applied to determine AA in single rat hepatocytes and the amount of AA in seven rat hepatocytes ranged from 16 to 62 fmol. The above results demonstrated that CE coupled with ECL is convenient, sensitive, and will become an attractive alternative method for single-cell analysis.
Key Laboratory of Eco-chemical Engineering, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao, China.
Xuemei
X
Sun
Yan
Y
Niu
Sai
S
Bi
Shusheng
S
Zhang
D015082:2,2'-Dipyridyl / Q000031:analogs & derivatives; D000818:Animals; D001205:Ascorbic Acid / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods*; D022781:Hepatocytes / Q000737:chemistry*; D008163:Luminescent Measurements / Q000379:methods; D008297:Male; D009942:Organometallic Compounds; D051381:Rats; D012428:Ruthenium
D009942:Organometallic Compounds; C103599:bis(bipyridyl)ruthenium(II); D015082:2,2'-Dipyridyl; D012428:Ruthenium; D001205:Ascorbic Acid