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(a) The linear calibration curve in Figure 4-13 isy=0.01630(±0.00022)x+0.0047(±0.0026)withsy=0.0059. Find the quantity of unknown protein that gives a measured absorbance of when a blank has an absorbance of 0.095

(b) Figure 4-13 has n=14 calibration points in the linear portion. You measure k=14replicate samples of unknown and find a mean corrected absorbance of 0.169 Find the standard uncertainty and 95%confidence interval for protein in the unknown.

Short Answer

Expert verified

(a) The amount of protein is 10.1μg.

(b) The standard uncertainty uxis 0.2015μgand 95% confidence interval is 10.08μg±0.44.

Step by step solution

01

Formula of Standard Uncertainty

The problem is based on the Least square method.

The formula of standard uncertainty is as follows:

ux=Sy|m|1k+1n+(y-y)2m2∑(xi-x)2.

Here, the standard uncertainty is ux, the standard deviation of y is sy , the total number of measurement is n, the value of the measurement of the unknown is k, the slope is m, the mean of y measurements is y, the mean of x measurements is x.

02

Calculate the corrected absorbance and the amount of protein

(a)

The formula of absorbance isabsorbance=m×(μ²µofprotein)+b.

Rearrange the formula for μg of protein as follows:

μgof protein =absorbance-bm.

Check page number 85 (figure 4.13 ) in the textbook for the values of the slope and the intercept.

The value of the slope is 0.0163 and the intercept is 0.0047.

Calculate the corrected absorbance as follows:

Subtract the absorbance of the blank from the absorbance of the measured to calculate the corrected absorbance:

Corrected absorbance=0.264-0.095

=0.169.

μgof protein =0.169-0.00470.0163

μgof protein =0.16430.0163

μgof protein =10.079μg

So, the amount of protein is10.1μg.

03

Calculate ∑ (xi-x)2 and standard uncertainty

(b)

Use standard uncertainty formula to calculateux

Refer to figure 4-13 in the textbook (page number 85) for the value of sy.

The value of syis 0.0046.

Refer to figure 4-13 in the textbook (page number 87) for the values of xi, y, x, andy.The values of localid="1663566988931" xare 0,0,0,5.0,5.0,5.0,10.0,10.0,10.0,15.0,15.0,20.0,20.0,20.0and xis 9.643μg. The values of y is 0.302 and yis 0.1618.

Calculate∑xi-x2value as follows:

∑(xi-x)2=(0-9.643)2+(0-9.643)2+(0-9.64.643)2++(10-9.643)2+(10-9.643)2+(10-9.643)2+(20-9.643)2+(20-9.643)2+(20-9.643)2∑(xi-x)2=92.99+92.99+92.99+21.56++28.7+28.7+107.27+107.27+107.27∑(xi-x)2=723.25.

Replace 0.0163 for m,0.0059 for sy, 4 for k, 14 for n, 724 for ∑(xi-x)2, 0.302 for y, and 0.1618 for yand calculateux.

ux=0.0050.016314+114+0.302-0.16110.01632723.25=0.3114+114+0.01970.1922=0.31×0.65.nux=0.31×0.65ux=0.2015.

So, the standard uncertainty uxis±0.2015.

04

Confidence Interval

The formula to find the confidence interval for an unknown measurement x is as follows:

Confidence intervals for x=±tux.

Here, the value of Student's is tand the standard uncertainty isux.

For the degree of freedom 12, the value of t for the 95% confidence interval is $2.179.

From the solution of part (b),uxis 0.2015.

Replace 0.2015 for uxand 2.179 for tand calculate the confidence interval.

Confidence intervals forx=±2.179×0.2015.

Confidence intervals for x=±0.439.

Thus, the standard uncertainty uxis 0.2015 μgand the 95% confidence interval is10.08μg±0.44.

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