ection: Name: 1.6mL of a solution containing both the oxidized and reduced form

Question

ection: Name: 1.6mL of a solution containing both the oxidized and reduced form of nicotinamide adenine inucleotide (NAD and NADH) was placed in a 1.6cm spectrophotometer cuvette. The absorbance at 340nm (which measures NADH) was 0.57. The absorbance at 260nm (which measures both NAD and NADH) was 1.90. The molar extinction coefficient of NADH at 340nm is 6220 L/(mol*cm), and the mola extinction coefficient of both compounds at 260nm is 18000 L/(mol"cm). Calculate the molar concentrations (in molarity) of NAD and NADH in the mixed solution X:.om , As at340--p 0bs ara60-1.90. calalet conc. NAD , NADH C Abs EX 1.6x622 0 1:00 6.

Explanation / Answer

Given data:

Path length of cuvette = 1.6

A340 of NADH = 0.57

A260 of NAD = 1.90

€ of NADH at 340 = 6220 L/ mol* cm

€ of NAD at 260 = 18000 L/mol*cm

Conc. of NADH and NAD = ?

formula is A = €BC

C = A / b * €

Thus, conc. of NADH = 0.57 / 1.6 * 6220 = 5.72 * 10^-5 M

Conc.of NAD = 1.90 / 1.6 * 18000 = 6.59 * 10^-5 M

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