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11 Reactions were conducted at 37 °C in 50 mM sodium phosphate buffer (pH 7.5) and samples removed at selected times. HPLC was done using a Rainin Dynamax C-18 column (4.6 × 150 mm, 100 Å, 5 μm, Varian Instruments) with a linear gradient of 5-80% acetonitrile (v/v) in 50 mM ammonium acetate (pH 7.0) over 30 min. Products were monitored at 260 nm. Initial velocities were determined by integration of isoniazid peak areas.
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