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Cypex CYP2D15
QC
Assays
Activity: Bufuralol
1'-hydroxylase
Incubation
conditions
Assay
buffer: 0.05 M potassium phosphate pH 7.4, 5 mM MgCl2
Incubation volume: 200 µl
Stop reagent: 70% perchloric acid (10 µl)
Internal standard: none
HPLC
conditions
Column:
HyperClone BDS-C18 (3 µm) 125 x 4 mm
Temperature: 30°C
Mobile phase: 10 mM perchloric acid/water/acetonitrile
20/56/24 (% v/v/v) initial
(gradient separation)
Flow rate: 1.0 ml/min
Run time: 12.5 min
Injection volume: 10 µl
Detection: fluorescence, l(ex)
= 252 nm; l(em)
= 302 nm
Retention times:
1'-hydroxybufuralol
2.8
min
bufuralol
5.4
min
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, l(ex)
= 270 nm; l(em)
= 312 nm
Retention times:
dextrorphan 2.4
min
dextromethorphan 10.9
min
Made
under licence from BTG International Ltd (AU730155, EP0914446,
US6566108 and other patents pending).
United States Patent Nos. 5,420,027 or 5,240,831, Canada Patent No.
2100245 and other patents pending.
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