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2, 4, 8, and 15 min for CYP2D6), 10 L aliquots of the preincubation mixture were transferred into secondary reaction tubes containing CYP-specific probe substrates (50 M bupropion for CYP2B6 and 5 M bufuralol for CYP2D6), 1 mM NADPH, 10 mM MgCl 2 , and 50 mM potassium phosphate buffer (pH 7.4) to a final volume of 100 L