Table 5.

Monitoring peroxide production and reduced nucleotide consumption with controls.

Rxn.	Peroxide at early time (nmoles/ml)		Peroxide at later time (nmoles/ml)		Consumption rate (nmoles/ml/min)
	NADPH (10 min)	NADH (16 min)	NADPH (30 min)	NADH (33 min)	NADPH	NADH
CYP + Diclof	0	0	0.89 ± 0.2	1.3 ± 0.4	0.13 ± 0.06	0.10 ± 0.01
		3.8 ± 1.9		4.9 ± 0.9		nd
CPR + Diclof	1.96 ± 0.1*	1.0 ± 0.1	5.74 ± 0.4*	1.8 ± 0.7	0.74 ± 0.03*	0.12 ± 0.01
		9.0 ± 0.5		14.2 ± 0.1		nd
CPR + CYP	2.64 ± 0.2	2.9 ± 0.1	6.32 ± 0.1	1.82 ± 0.2	0.90 ± 0.04	0.27 ± 0.03
		12.7 ± 0.7		21.4 ± 0.3		nd
CPR + CYP + Diclof	3.44 ± 0.1*	2.1 ± 0.3	6.23 ± 0.1*	2.0 ± 0.5	1.08 ± 0.09*	0.37 ± 0.03
		11.2 ± 0.5		18.1 ± 1.1		nd

[CPR] = 100 nM, [CYP2C9] = 100 nM and [Diclof] = 100 μM. (The upper row in NADH reactions had 135 μM NADH and the lower one had 1350 μM NADH.) Rate of NADH depletion was determined from the slope of linear fit of 340 nm OD at 0, 90 and 180 min. The controls with CYP or CPR also included Diclof. For NADPH reactions, the initial concentrations of components were–[CYP2C9] = 100 nM, [CPR] = 100 nM, [Diclof] = 200 μM, and [NADPH] ~200 μM. NADPH depletion rate was determined from the slope of linear fit of 340 nm OD at 16, 26 and 36 min. The controls with CYP or CPR did not include Diclof. nd: not determined,

^*data previously reported in our work, given here for comparison (Manoj et al., 2010b).