A.

Void Volume Determination

B. Protein Standard Solution

Void Volume Determination

Potein Standard Solution

0.14

0.6

0.12

0.5
Absorbance

Absorbance

0.1
0.08
0.06
0.04

0.3
0.2
0.1
0

0.02

-0.1 0

0
-0.02

0.4

10
20
30
Equivalent Volume

Standards Used

(Da)

B-amylase
Alcohol anhydrase
BSA
Carbonic anhydrase
Cytochrome C

200,000
150,000
66,000
29,000
12,400

20

80

40

Elution Volume
(Ve) (mL)
11
18
29
40
51

Void Volume (Vo)

(mL)
5
5
5
5
5

Calibration Curve

Ve/Vo

Log MW

2.2
3.6
5.8
8
10.2

5.30103
5.176091
4.819544
4.462398
4.093422

C. Sample (Casein and Albumin)

Sample (casein and albumin)

Calib curve
3

6
5
4
3
2
1
0

Absrobance

log MW

40
60
Equivalent Volume

y = -0.1543x + 5.6899
R = 0.9947

10

Ve/Vo

Sample
Albumin
Casein

Ve/Vo
8.4
7.4

12

2
1
0
0

10

-1

20

30

Equivalent Volume

logMW
4.39378
4.54808

MW
24761.67
35324.82

40

50