Figure 3.

Products resulting from the CYP76B1-dependent metabolism of chlortoluron and isoproturon. Approximately 8 pmol of CYP76B1 in WAT11 yeast microsomes was incubated for 60 min at 30°C with 500 μm substrate, 500 μm NADPH, 1 mm Glc-6-P, and 0.5 unit of Glc-6-P dehydrogenase in 200 μL of 0.1 m sodium phosphate, pH 7.4. Twenty microliters of the acidified and centrifuged incubation medium was directly analyzed by liquid chromatography-MS using a gradient of 3% to 60% B (100% acetonitrile, 0.08% TFA) in A (water, 0.1% TFA) in 65 min. Absorbance was monitored at 214 nm.