ALKALINE PHOSPHATE EXPERIMENT
Table: Different Temperature with same volume of Enzyme.
Wavelength : 415nm
Temperature : 28c, 33c
Volume of Enzyme : 0.5ml
Time
0.5 ml enzyme/
Absorbance
Time
0.5 ml Enzyme
Absorbance @ 33C
0
0.065
0
0.068
2
0.077
2min
0.085
4
0.102
4min
0.098
6
0.115
6min
0.112
8
0.135
8min
0.136
10
0.151
10min
0.151
12
0.175
12min
0.189
17
0.203
14min
0.215
27
0.303
16min
0.250
37
0.418
18min
0.275
47
0.492
20min
0.350
57
0.572
22min
0.375
67
0.572
24min
0.428

26min
0.465

28min
0.500

30min
0.550

32min
0.595

34min
0.660

36
0.770

38
0.780

40
0.820

46
0.860

Table 2 : Same Temperature but different Enzyme levels
Temperature : 28c
Volume of Enzyme : 1.5 ml and 0.5 ml
Wavelength : 415nm
Time
0.5 ml enzyme/
Absorbance
Time
1ml Enzyme
Absorbance @ 28C
Time
1.5ml Enzyme
Absorbance
0
0.065
0
0.028
0
0.236
2
0.077
2
0.095
2
0.334
4
0.102
4
0.170
4
0.428
6
0.115
6
0.224
6
0.520
8
0.135
8
0.286
8
0.696
10
0.151
10
0.337
10
0.875
12
0.175
12
0.390
12
0.985
17
0.203
14
0.446
14
1.045
27
0.303
16
0.505
16
1.220
37
0.418
18
0.566
18
1.440
47
0.492
20
0.635
20
1.640
57
0.572
22
0.682
22
2.100
67
0.572
24
0.756
24
2.440

0.830
26

0.900
28

0.975
30

1.009
32

1.018
34

1.022
36

Graph 2: Time vs Absorbance at 415 nm at different enzyme concentration at same (28 ° C) temperature At higher concentration of enzyme the more absorbance observed. The lesser the concentration the lesser absorbance observed. According to the above graph concentration of enzyme is directly proportional to absorbance.

Post Lab Questions:
Comment on the accuracy of the data from other students.
The accuracy of the data was varying according to the amount of enzymes used and the temperature as a variable. If the amount of the enzymes is 1ml or 0.5 ml the reaction was slower and it takes some time to developed the color. Even the time was one of the important factors in