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Cynomolgus monkey CYP2C8 (CYP2C20) QC Assays

Activity: Paclitaxel 6α-hydroxylase

Incubation conditions
Assay buffer: 0.1 M potassium phosphate pH 7.4, 5 mM MgCl2
Incubation volume: 200 µl
Stop reagent: acetonitrile (100 µl)

HPLC conditions
Column: Hypersil BDS-C18 (3 µm) 100 x 4 mm
Temperature: 40°C
Mobile phase: 20 mM potassium phosphate pH 7/acetonitrile 57.5/42.5 (v/v) (isocratic)
Flow rate: 1.0 ml/min
Run time: 8.5 min
Injection volume: 75 µl
Detection: UV, λ = 230 nm

Retention times:
6α-hydroxypaclitaxel 4.5 min
paclitaxel 18.5 min

The substrate, paclitaxel, and metabolite, 6α-hydroxypaclitaxel, are both available from Cypex Ltd.

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