Activity: Dextromethorphan O-demethylase
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: Hypersil BDS-C18 (3 µm) 100 x 4 mm
Temperature: 30°C
Mobile phase: 0.05% orthophosphoric acid/acetonitrile 78/22 (v/v) (isocratic)
Flow rate: 1.0 ml/min
Run time: 13.5 min
Injection volume: 50 µl
Detection: fluorescence, λ(ex) = 270 nm; λ(em) = 312 nm
Retention times:
dextrorphan 2.4 min
dextromethorphan 10.9 min