Table 1

Incubation conditions, HPLC assay conditions, and K_m values for cDNA-expressed P450 isoform (Supersomes) activities

P450 Isoform	Substrate	Enzyme (pmol P450/inc)	Metabolite	Internal Standard (200 μl of Acetonitrile)	Mobile Phase Acetonitrile/10 mM Potassium Phosphate Buffer (pH 8.0)	Excitation	Emission
						nm
CYP1A2	2.5 μM CEC	0.5	CHC	0.5 μM HMC	0–10 min; 20:80–70:30 linear gradient	408	450
CYP2A6	0.5 μM Coumarin	0.4	HC	0.1 μM HMC	0–5 min; 20:80–30:70 linear gradient, 5–10 min; 30:70	368	456
CYP2C8	0.5 μM DBF	2.5	FL	40 nM DCFL	0–5 min; 15:85–32:68 linear gradient, 5–12 min 70:30	490	525
CYP2C9	35 μM MFL	2.5	FL	40 nM DCFL	0–10 min; 15:85–35:65 linear gradient	490	525
CYP2C19	1.5 μM MFL	2.5	FL	40 nM DCFL	0–10 min; 15:85–35:65 linear gradient	490	525
CYP2D6	5.0 μM MAMC	2.5	HAMC	0.1 μM HC	0–10 min; 10:90–30:70 linear gradient	370	470
CYP2E1	100 μM MFC	5.0	HFC	0.25 μM HC	0–10 min; 20:80–70:30 linear gradient	385	502
CYP3A4	0.5 μM DBF	0.5	FL	40 nM DCFL	0–5 min; 15:85–32:68 linear gradient, 5–12 min 70:30	490	525

CHC, 3-cyano-7-hydroxycoumarin; HC, 7-hydroxycoumarin; FL, fluorescein; HAMC, 7-hydroxy-4-(aminomethyl)coumarin; HFC, 7-hydroxy-4-(trifluoromethyl)coumarin; HMC, 7-hydroxy-4-methylcoumarin; DCFL, 2,7-dichlorofluorescein.