On-line drug metabolites generation and their subsequent target analysis by capillary zone electrophoresis with UV-absorption detection.
This study presents the in-capillary enzymatic biotransformation of dextromethorphan, an antitusive drug and opioid receptor antagonist, and subsequent electrophoretic separation of its products. The study includes the optimization of separation parameters to fulfill the requirements of an online microreaction. The analyses were performed in a bare fused-silica capillary using 100 mM sodium tetraborate (pH 10.0) mixed with linear polyacrylamide (20%, v/v) and 2-propanol (10%, v/v). This BGE was suitable for monitoring both off-line and in-capillary incubations. The partial filling technique enabled the enzymatic reaction to be carried out in its optimal environment (20 mM sodium phosphate, pH 7.4). Finally, in-capillary microreaction in the presence of cytochrome P450 3A4 gave satisfactory outcomes.
Department of Biochemistry, Faculty of Science, Masaryk University, Brno, Czech Republic. zeisbergerova@mail.muni.cz
Marta
M
Zeisbergerová
Roman
R
Řemínek
Aleš
A
Mádr
Zdeněk
Z
Glatz
Jos
J
Hoogmartens
Ann
A
Van Schepdael
D015374:Biosensing Techniques; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D003915:Dextromethorphan / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008862:Microsomes, Liver / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet
D003915:Dextromethorphan; D003577:Cytochrome P-450 Enzyme System