Enantioselective binding analysis of verapamil to plasma lipoproteins by capillary electrophoresis-frontal analysis.
Capillary electrophoresis coupled with frontal analysis was applied to the study of enantioselective binding of verapamil (VER) to plasma lipoproteins. The drug-lipoprotein mixed solution, which had been in the binding equilibrium, was hydrodynamically introduced into a non-coated fused-silica capillary. Since VER is positively charged in the neutral run buffer (pH 7.4), the unbound VER enantiomers migrated toward the cathodic end much faster than negatively charged lipoproteins and their bound forms. Once unbound VER migrated apart from lipoprotein, the bound VER was quickly released from the protein to maintain the binding equilibrium. Thus, VER migrated as a zone through the capillary and gave a trapezoidal peak with a plateau region on the electropherogram. The VER concentration in this plateau region was equal to the unbound VER concentration in the initial sample solution. It was found that the bindings of VER to high-density lipoprotein (HDL), low-density lipoprotein (LDL) and oxidized LDL were not site-specific and not enantioselective. Partition-like binding to lipid part of these lipoproteins seemed to be dominant. The total binding affinities of LDL to VER were about seven-times stronger than those of HDL, and the oxidation of LDL by copper ion enhanced the binding affinities significantly.
Mohamed
N A
NA
Graduate School of Pharmaceutical Sciences, Kyoto University, Japan.
Kuroda
Y
Y
Shibukawa
A
A
Nakagawa
T
T
El Gizawy
S
S
Askal
H F
HF
El Kommos
M E
ME
D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D008074:Lipoproteins / Q000097:blood*; D008297:Male; D013237:Stereoisomerism; D014700:Verapamil / Q000097:blood*
D008074:Lipoproteins; D014700:Verapamil