HW 9 - SP6 - Approximate Analysis of an Elution Chromatograph - 6 pts
Three proteins, A, B and C, are eluted through a pilot plant chromatographic column. The outlet concentration as a function of time was determined using an ultraviolet spectrophotometer. The results appear in the graph shown below.
a.) Estimate y0, t0 and σt0 for each species.
b.) Determine the yield and purity of species B in a fraction collected between t = 30 min and t = 40 min.
c.) What is the concentration of species B in this fraction ?
Elution Chromatograph
a.) Estimate y0, t0 and σt0 for each species.
b.) Determine the yield and purity of species B in a fraction collected between t = 30 min and t = 40 min.
c.) What is the concentration of species B in this fraction ?
Elution Chromatograph
posted by Dr. B at 8:39 AM
3 Comments:
Here is the rule of thumb you need for this problem.
The dimensional standard deviation...sigm*t0 is approximately equal to half the width of the peak evaluated at half the height of the peak.
What equation do we use for the concentration of B? I found the purity and yields just fine, but I'm stuck on this part. Help...guidance...please
almost:
[B] = MB / Vol in the cut or fraction from t' to t
[B] = [INT{Fy dt} from t' to t] / [INT{F dt} from t' to t]
Yield = [INT{Fy dt} from t' to t] / [INT{Fy dt} from -infinity to infinity ]
Therefore, [B] = Yield * [INT{Fy dt} from -infinity to infinity ] / [INT{F dt} from t' to t]
Canel F because it is a constant.
[B] = Yield * [INT{y dt} from -infinity to infinity ] / [INT dt} from t' to t]
[INT dt} from t' to t] = t-t'
[B] = Yield * [INT{y dt} from -infinity to infinity ] / (t-t')
You can integrate the Gaussian peak to get: [INT{y dt} from -infinity to infinity ] = y0*sigma*t0*sqrt(2*pi)
We can also show that this is true without integration, but it is hard to do in text here. Instead, I will email a PPT file to everyone showing why [INT{y dt} from -infinity to infinity ] = y0*sigma*t0*sqrt(2*pi)
Long comment !?
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