A sample of RNA is hydrolyzed and separated into three fractions by column chrom

Question

A sample of RNA is hydrolyzed and separated into three fractions by column chromatography. Two of the three fractions are pure nucleotides, but the third contains both adenylic and guanylic acids. At pH 7.0, the absorbance of the mixture is 0.305 at 280 nm and 0.655 at 250 nm in 1-cm cells. The molar extinction coefficients for each pure component at pH 7.0 are: Calculate the concentration of adenylic and guanylic acids in the 1-cm cell What is the ratio of A to G nucleotides present in the original RNA sample?

Explanation / Answer

Abs=Ecl

At 280 nm abs of mixture =2300*CA  *1+9300*CG *1

let Ca X

Let CG Y

0 .305 =2300X+9300Y (EQ-1)

0.655 =12300X+15700Y (EQ-2)

Solving both eq

X=0.0000219439 (adenalyic acid conc)

Y=0.000024528 (Guanlylica acid conc)

AG Ratio 0.895

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