Assay of bradykinin metabolites in human body fluids by CE-LIF coupled with transient ITP preconcentration.
A CE with LIF detection was developed for the separation and determination of three bradykinin (BK) metabolites: BK2-9, BK1-7 and BK1-5. BK fragments were derivatized with 5-(4, 6-dichloro-s-triazin-2-ylamino) fluorescein before CE-LIF analysis. Eighty millimolar of Tris-H3PO4 (pH 9.0) was selected as the derivatization media. Three BK fragments were baseline separated within 10 min by using 0.2 M TAPS-Tris buffer (pH 8.5) as the running buffer. Meanwhile we have also developed a simple, quick, and sensitive on-column transient ITP preconcentration for CE-LIF detection of three BK fragments. Ten millimolar of Tris-HCl (pH 9.0) was chosen as the leading electrolyte and 0.2 M TAPS-Tris (pH 8.5) containing 10 mM 1-butyl-3-methylimidazolium tetrafluoroborate as the terminating electrolyte and also served as the running buffer during CZE separation. By using this transient ITP coupled with CE-LIF, concentration detection limits (S/N=3) for BK2-9, BK1-7 and BK1-5 were 0.3, 0.1 and 0.1 pmol/L, respectively. This method has been applied to the assay of human saliva and plasma samples with satisfactory results.
Chen
Ying
Y
Ministry of Education Key Laboratory of Analysis and Detection Technology for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou, Fujian, P. R. China.
Zhang
Lan
L
Xu
Liangjun
L
Lin
Jin-Ming
JM
Chen
Guonan
G
D001920:Bradykinin / Q000032:analysis* / Q000097:blood / Q000378:metabolism; D004586:Electrophoresis / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005452:Fluoresceins / Q000737:chemistry; D005453:Fluorescence; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000737:chemistry; D007834:Lasers; D016014:Linear Models; D015203:Reproducibility of Results; D012463:Saliva / Q000737:chemistry* / Q000378:metabolism; D012680:Sensitivity and Specificity; D014227:Triazines / Q000737:chemistry; D014325:Tromethamine / Q000737:chemistry
C525963:1-butyl-3-methylimidazolium; C508435:5-(4,6-dichloro-s-triazin-2-ylamino) fluorescein; D005452:Fluoresceins; D007093:Imidazoles; D014227:Triazines; D014325:Tromethamine; D001920:Bradykinin