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(a)Xanthine absorbs at 260 nm with an extinction coefficient of 3.85 x 103 M-1 c

ID: 488047 • Letter: #

Question

(a)Xanthine absorbs at 260 nm with an extinction coefficient of 3.85 x 103 M-1 cm-1 . An absorbance of 0.72 was recorded in a 1 cm cuvette containing 3 mL of solution. Stock solution (100 L ) had been added to 2.9 mL diluents to prepare sample for the measurement. Calculate the xanthine concentration in the stock solution

(b) The 280 nm absorbance of a solution of a pure protein was 0.86. The protein content, determined by BCA method, was 510 g per mL. If the molecular weight of the protein is 40,000, what is the molar (M) extinction coefficient? The mM extinction coefficient?

Explanation / Answer

a)

From Beer Lambert Law

A=Ecl

A = Absorbance = 0.72

E = 3.85 x 10^3 M-1cm-1 = Molar absorptivity

l = 1 cm = Path length

c = Concentration

Hence , c = A/El = 0.72 / 3.85 x 10^3 x 1 =1.87 x 10^-4 M

Hence,

S1 = 3 x1.87 x 10^-4 / 10^-4 =5.61 M

Hence stock solution concentration is 5.61 M