So the absorbance spectrum below is of a solution of human hemoglobin in water.

Question

So the absorbance spectrum below is of a solution of human hemoglobin in water. The spectrum in red is of the hemoglobin in its deoxy form. The spectrum in blue is of teh same solution of hemoglobin, but following exposure of the solution to air.

a)Circle all the isosbestic points that you see on the spectra. How many are there, and at approximately what wavelengths do they occur?

b) Calculate the concentration of the hemoglobin solutions given that the molar extinction coefficient of oxyhemoglobin at 500 nm is roughly 21,000 M^-1 cm^-1, and the gram molecular weight is 64,500.

c) In another experiment, rather than exposing the deoxyhemoglboin to air, a small volume of air was injected into the sealled cuvette holding the hemoglobin. The absorbance of the hemoglobin at 560 nm decreased from .54(absorbance of the deoxy-H at 560nm) to .46. The absorbance fo the oxyhemoglobin at 560 nm was .33. What fraction of the hemoglobin was complexed with oxygen?

d) after another injection of a small amount of air, the absorbance at 560 nm decreased further to .40. After teh second injection of air, what was the fraction of hemoglobin complexded with oxygen.

Already got through with part A so no need to answer that one. I need the rest of them and please show work! I'm running short on time so im leaving this one up to you guys. Thanks...

So the absorbance spectrum below is of a solution of human hemoglobin in water. The spectrum in red is of the hemoglobin in its deoxy form. The spectrum in blue is of teh same solution of hemoglobin, but following exposure of the solution to air. Circle all the isosbestic points that you see on the spectra. How many are there, and at approximately what wavelengths do they occur? Calculate the concentration of the hemoglobin solutions given that the molar extinction coefficient of oxyhemoglobin at 500 nm is roughly 21,000 M^-1 cm^-1, and the gram molecular weight is 64,500. In another experiment, rather than exposing the deoxyhemoglboin to air, a small volume of air was injected into the sealled cuvette holding the hemoglobin. The absorbance of the hemoglobin at 560 nm decreased from .54(absorbance of the deoxy-H at 560nm) to .46. The absorbance fo the oxyhemoglobin at 560 nm was .33. What fraction of the hemoglobin was complexed with oxygen? after another injection of a small amount of air, the absorbance at 560 nm decreased further to .40. After teh second injection of air, what was the fraction of hemoglobin complexded with oxygen. Already got through with part A so no need to answer that one. I need the rest of them and please show work! I'm running short on time so im leaving this one up to you guys. Thanks...

Explanation / Answer

sorry i am unable to upload the images...server is not responding

part b) we have

A=e*c*l

considering 1 cm cuvette, l=1 cm from graph A=.21 at 500 nm

given e =21000 /M cm ...w e get

c=0.21/21000 =0.01 milli M

c) percentage from the given values can be calculated as follows

x=(54-46)/(54-33) *100 =42.1 %

similarly part d)

x= (54-40)/(54-33)*100 =73.68 %

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