Activity: Bupropion hydroxylase
Incubation conditions
Assay buffer: 0.05 M potassium phosphate pH 7.4, 5 mM MgCl2
Incubation volume: 200 µl
Stop reagent: 1 M hydrochloric acid (20 µl)
Internal standard: none
HPLC conditions
Column: HyperClone BDS-C18 (3 µm) 125 x 4 mm
Temperature: 30°C
Mobile phase: 100 mM potassium phosphate pH 2/acetonitrile/water 20/16/64 (v/v/v) (isocratic)
Flow rate: 1.0 ml/min
Run time: 17.5 min
Injection volume: 100 µl
Detection: UV, λ = 196 nm
Retention times:
hydroxybupropion 6.3 min
bupropion 12.4 min