An assay for angiotensin-converting enzyme using capillary zone electrophoresis.
A sensitive and rapid method was developed for angiotensin-converting enzyme (ACE) activity determination by capillary zone electrophoresis. Hippuryl-l-histidyl-l-leucine, a synthetic tripeptide, was used as the ACE-specific substrate. Capillary zone electrophoresis was employed to separate the products of the enzymatic reaction and the ACE activity was determined by quantification of hippuric acid, a result of the enzymatic reaction on the tripeptide. The capillary electrophoresis was performed in a 27 cm x 75 micrometer i.d. fused-silica capillary using 200 mM boric acid-borate buffer (pH 9.0) as a run buffer with an applied voltage of 8.1 kV at a capillary temperature of 23 degrees C. The electrophoresis was monitored at 228 nm. Each electrophoretic run requires only a nanoliter of the enzymatic reactant solution, at only 6 min, rendering a powerful tool for the ACE assay.
Zhang
R
R
Institute of Biotechnology, Fuzhou University, Fujian, 350002, People's Republic of China.
Xu
X
X
Chen
T
T
Li
L
L
Rao
P
P
D002138:Calibration; D019075:Electrophoresis, Capillary / Q000379:methods*; D006626:Hippurates / Q000737:chemistry; D007700:Kinetics; D007703:Peptidyl-Dipeptidase A / Q000032:analysis*; D012015:Reference Standards
D006626:Hippurates; D007703:Peptidyl-Dipeptidase A; C030514:hippuric acid