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Cypex cynomolgus monkey CYP2C76 QC Assays

Activity: Tolbutamide 4-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)

HPLC conditions
Column: HyperClone ODS (5 µm) 250 x 4 mm
Temperature: 30°C
Mobile phase: Water/100 mM potassium phosphate pH 2/methanol/acetonitrile, 50/10/35/5 (v/v/v/v) (initial conditions)
Run type: gradient (5 - 25% acetonitrile over 4 min)
Flow rate: 1.0 ml/min
Run time: 16 min
Injection volume: 100 µl
Detection: UV,
l = 230 nm
Retention times:

      4-hydroxytolbutamide        5.9 min
     (carboxytolbutamide          6.8 min)
      tolbutamide                     9.1 min

The CYP2C76 substrate tolbutamide and metabolite 4-hydroxytolbutamide are available from Cypex.

 

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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