EXER 5.3.

RAW DATA

Table 5.3.1. Data for the measurement of column volume (VT) and flow rate
Parameter Value
Height of gel bed, cm 17.5
Inner diameter, cm 1.3
Radius, cm 0.65
Column Volume, cm3 23.22815068
Flow rate, mL/min 1.23954137

Table 5.3.2. Data for the elution volume and absorbance of protein standards.
Test Tube Elution Absorbance Test Tube Elution Absorbance
No. Volume, mL (A280) No. Volume, mL (A280)
1 2 0.0000 9 18 1.4240
2 4 -0.0170 10 20 0.4766
3 6 0.0021 11 22 0.6425
4 8 0.1273 12 24 0.1543
5 10 0.0385 13 26 0.1703
6 12 0.1597 14 28 0.0471
7 14 0.2952 15 30 0.1862
8 16 1.1174

Table 5.3.3. Molecular weight and elution volumes of protein standards.

Molecular Weight Elution volume (Ve),
Protein Standard log MW
(MW), kDa mL
myoglobin 16.7 1.222716471 18
blue dextran 2000 3.301029996 8
bromcresol purple 0.540 -0.267606240 22

Table 5.3.4. Linear regression parameters for the plot of log MW versus elution volume.
Parameter Value
Slope, mL-1- -0.245850428
y - intercept 5.35232025
R -0.989109016
R2 0.978336645

Table 5.3.5. Data for the elution volume and absorbance of egg albumin isolate.
Test Tube Elution Absorbance Test Tube Elution Absorbance
No. Volume, mL (A280) No. Volume, mL (A280)
1 2 0.016 9 18 0.077
2 4 0.042 10 20 0.034
3 6 3.135 11 22 0.014
4 8 2.913 12 24 0.012
5 10 1.474 13 26 0.016
6 12 0.421 14 28 0.008
7 14 0.643 15 30 0.007
8 16 0.201
Table 5.3.6. Data for the distribution coefficients of protein standards and egg albumin.
Elution Volume Void Volume Total Volume
Protein Kav
(Ve), mL (V0), mL (VT), mL
myoglobin 18 0.6566785561
blue dextran 8 0
bromcresol 22
8 23.22815068 0.9193499785
purple
egg albumin 14
03940071336
isolate

Table 5.3.7. Comparison of the experimental and theoretical values of the molecular weight
of egg albumin isolate.
Parameter Value
Experimental molecular weight of egg
81.2
albumin isolate (MWexp), kDa
Theoretical molecular weight of egg
45.0
albumin isolate (MWtheo), kDa
Percent error (%) 80.4

EXER 5.4.

Table 5.4.1. Retention factor of different proteins based on the obtained chromatogram.
Distance traveled,
Protein MW, kDa log MW Rf
cm
phosphorylase b 97 4.986771734 2.9 0.263636364
albumin 66 4.819543936 4.3 0.390909091
ovalbumin 45 4.653212514 6.0 0.545454545
carbonic anhydrase 30 4.477121255 6.7 0.609090909
trypsin inhibitor 20.1 4.301029996 8.3 0.754545455
- lactalbumin 14.4 4.158362492 9.7 0.881818182
Distance traveled by
11.0
tracking dye, cm

Table 5.4.2. Data for the determination of molecular weight of protein components of
samples.
Distance
Component/Sample Rf MW, Da
traveled, cm
albumin 4.3 0.390909091 66
ovalbumin 6.0 0.545454545 45
trypsin inhibitor 8.3 0.754545455 20.1
- lactalbumin 9.7 0.881818182 14.4
Distance traveled by
11.0
tracking dye, cm

Parepareho lang naman distance di na kailangan iinterpolate (duh)