Seung Soo (Jason) Lee 002213-065
Internal Assessment – Investigating the Relationship between Concentration of Sodium Chloride and the Rate of Reaction of Enzyme Amylase
Research Question: How will changing the percentage of sodium chloride concentration affect the rate of reaction of enzyme amylase, measured using the absorbance of starch and iodine with a spectrophotometer.
Introduction: Amylase is an enzyme that is involved in the human digestive process. Found in both the human pancreas and the human saliva, amylase breaks down starch into sugar so that large molecules can be easily digested1. Like all enzymes, amylase must be kept in a certain condition in order to function properly. In this experiment, the effect of sodium chloride concentration on the rate of reaction of amylase will be investigated with the use of starch and iodine. When starch is mixed with iodine, the coils of beta amylose molecules found in starch trap iodine, causing the mixture to turn into a shade of blue-black. 2 When starch is broken down into glucose, however, the monosaccharide does not react with iodine. Therefore, glucose does not change color even when it’s mixed with iodine. Correspondingly, when drops of amylase are inputted into a blueblack mixture of starch and iodine, the starch molecules will be broken down into glucose molecules, causing the mixture to turn colorless. Thus, the rate of reaction of amylase correlates to the absolute value of the rate of change in absorbance of the solution. A rapid decrease in the absorbance of the blue-black color equates to a high rate of reaction of amylase, whereas a slow decrease in absorbance signifies a low rate of reaction. In this experiment, an external variable of sodium chloride will be manipulated into the amylase enzyme to determine the effect the concentration of sodium chloride on the rate of reaction of amylase. Rate of Reaction = │
│
1
"Amylase." Wikipedia. N.p., n.d. Web. 12 Jan 2011. <http://en.wikipedia.org/wiki/Amylase>. 2
Senese, Fred. "How Does Starch Indicate Iodine?." N.p., 15 Feb 2010. Web. 6 Jan 2011. <http://antoine.frostburg.edu/chem/senese/101/redox/faq/starch -as-redox-indicator.shtml>.
Seung Soo (Jason) Lee 002213-065
Hypothesis: As aforementioned, amylase, like all enzymes, must be kept under a certain set of conditions in order to function properly. Factors such as pH level, temperature, and salt concentration could all denature the enzyme and decrease its activity. . When a substrate can no longer bind to the active site of an enzyme due to its conformational change, the enzyme activity and the rate of reaction of the enzyme drops significantly. For instance, a high concentration of sodium chloride would alter the electrostatic interactions between charged amino acids, causing conformational change in the enzyme and destroying its active site.3 Furthermore, the presence of sodium chloride will only have little impact on the enzyme structure unless the sodium chloride concentration is very high, when it could completely denature the enzyme. Therefore, an enzyme should experience an exponential decrease in its rate of reaction as the concentration of sodium chloride is increased Rate of Reaction of Amylase, Abss-1 Concentration of Sodium Chloride, %
Figure 1: Prediction of the Effect of Sodium Chloride Concentration on Rate of Reaction of Amylase Enzyme
Thus, the hypothesis for this experiment is that if the sodium chloride concentration is increased, then the rate of reaction of amylase will decrease. A high concentration of sodium chloride will denature the enzyme amylase and, as a result, it will no longer be able to break down starch into glucose. The figure above demonstrates that the average rate of change in absorbance will undergo an exponential decrease as the concentration of sodium chloride is increased.
3
"Rule of Protein Structure." N.p., n.d. Web. 6 Jan 2011. <http://users.rcn.com/jkimball.ma.ultranet/BiologyP ages/D/DenaturingProtein.html>.
Seung Soo (Jason) Lee 002213-065
Variables:
Variable
Description
Units / range
Method of Measuring / Manipulating
Independent
Concentration of sodium chloride
%
The independent variable will be manipulated by a process of serial dilution, from 20% concentration of sodium chloride to 10%, 10% to 5%, 5% to 1%, and 1% to 0.1%.
Dependent
Rate of reaction of amylase
│
│ (ΔAbss-1)
Controlled
This will be measured with a spectrophotometer and Logger Pro. Because amylase breaks down starch into glucose, and glucose does not react with iodine, the enzyme activity of amylase will lower the blue-black absorbance of starch+iodine. Therefore, the rate of decrease in absorbance over time correlates to the absolute value of the rate of reaction of amylase. The change in absorbance will be measured from 0-20 seconds, and the rate of reaction can be calculated by finding the slope of the absorbance vs. time graph. The uncertainty can be considered negligible.
Concentration of starch & iodine
%
This will be kept constant by using the same mixture created through steps 1-3 of procedures for every trial. (0.05% starch + 300μl of iodine)
Amount of solutions inside the cuvette
μl
For every trial, 2.5ml of starch & iodine solution and 500μl of sodium chloride & amylase solution is put inside the cuvette.
Temperature
°C
Temperature is kept constant by conducting the experiment at room temperature (about 25 °C) for every trial.
Table 1: List of Variables
Seung Soo (Jason) Lee 002213-065
Apparatus and Materials:
Electronic balance (±0.001g) 100 cm3 & 10 cm3 volumetric flasks 10 cm3 pipette (±0.02 cm3) 1000 μl & 50 μl micropipettes 3 cm3 cuvettes 2 cm3 micro tubes Microcentrifuge Five small beakers for serial dilution
1 medium sized beaker Sodium Chloride Iodine Starch Hot plate Vernier Spectrophotometer Logger Pro
Procedures: Preparation of 0.05% starch mixed with iodine 1. 0.05g of starch and 100cm3 of distilled water are poured into a medium sized beaker. 2. The beaker is placed on a hot plate, and then stirred several times using a plastic stirrer until a homogenous solution is made. 3. 300μl of iodine is put into the starch solution. It is stirred several times using a plastic stirrer until a blue-black solution is made.
Preparation of sodium chloride of various concentrations (serial dilution) 8 cm3 distilled water
3
5 cm distilled water
5 cm3
20%
5 cm3
10%
2 cm3
5%
9 cm3 distilled water
1 cm3
1%
Figure 2: Serial Dilution of Sodium Chloride Solution
0.1%
Seung Soo (Jason) Lee 002213-065 4. 2g of sodium chloride and 10cm3 of distilled water is poured into a small beaker. 5. The beaker is stirred several times using a plastic stirrer until a homogenous solution is made, creating a 20% sodium chloride solution. 6. 5 cm3 of the obtained solution is transferred into another small beaker using a 10 cm3 pipette, and another 5 cm3 of distilled water is added into the beaker. The beaker is then stirred using a stirrer until a homogenous solution is made, creating a 10% sodium chloride solution. 7. The serial dilution of sodium chloride is continued, according to the layout in figure 2, to obtain 5%, 1%, and 0.1% concentrations of sodium chloride solutions.
Conducting the experiment 8. The Vernier spectrophotometer is calibrated using distilled water. Then, the wavelength at which to measure the absorbance is determined using the maximum wavelength of the blueblack mixture of starch and iodine. 9. 450μl of 20% sodium chloride solution is put inside a micro tube using a 1000μl micropipette. 50μl of amylase solution is added into the micro tube using a 50μl micropipette. The micro tube is then placed inside a microcentrifuge so that the solution will mix together. 10. Step 9 is repeated three times for all concentrations of sodium chloride, creating three mixtures of sodium chloride and amylase for each of the five variables. 11. 2.5cm3 of the starch & iodine mixture is put into a 3cm3 cuvette using a micropipette. 500μl of the sodium chloride & amylase mixture is added into the cuvette using a micropipette. 12. The solution is squeezed in and out three times using the micropipette to ensure that amylase spreads throughout the starch solution. After mixing three times, the “start” button on Logger Pro is clicked. Steps 11 and 12 are performed with the cuvette placed inside the spectrophotometer to minimize error. 13. The rate of change in absorbance of the mixture is measured using Logger Pro for 20 seconds. 14. Steps 11-13 are repeated for triplicate trials for all five concentrations of sodium chloride.
Seung Soo (Jason) Lee 002213-065
Data Collection:
Qualitative Data:
Even with the naked eye, one could observe the disappearance of color inside the cuvette, from a dark, blue-black coloration to a clear, colorless state.
Quantitative Data:
*** Refer to the Appendix for a complete table of raw data from Logger Pro.
Seung Soo (Jason) Lee 002213-065
Data Processing: Rate of Decrease in Absorbance / ΔAbss-1
Sodium Chloride Concentration /% 20.0
Trial 1
Trial 2
Trial 3
-0.001240
-0.001280
-0.001223
10.0
-0.001517
-0.001299
-0.001402
5.0
-0.001812
-0.001523
-0.0012304
1.0
-0.001836
-0.001703
-0.001714
0.1
-0.001845
-0.001985
-0.001931
Control (no sodium chloride): -0.002830 Table 2: Rate of Decrease in Absorbance for All Trials5 Sodium Chloride Concentration / %
Calculation
Average Rate of Reaction (±Standard Deviation)6 / ΔAbss-1
20.0
0.001248 ± 0.000029
10.0
0.001406 ± 0.000109
5.0
0.001668 ± 0.000204
1.0
0.001751 ± 0.000074
0.1
0.001920 ± 0.000071
Control (no sodium chloride): 0.002830 ΔAbss-1 Table 3: Calculation of Average Rates of Reaction
**Because the rate of reaction must be a positive value, the average rate of reaction was taken as an absolute value. 4
This value was neglected in data processing because it was considered as an outlier.
5
The rate of decrease in absorbance was determined by finding the slope of absorbance vs. time graph
using linear regression on Logger Pro software. 6
The processing of standard deviation is shown in table 4
Seung Soo (Jason) Lee 002213-065
Data Presentation:
LEGEND Run 4: Control (no sodium chloride) Run 5: 20% sodium chloride Run 10: 10% sodium chloride Run 11: 5% sodium chloride Run 14: 1% sodium chloride Run 19: 0.1% sodium chloride
Figure 4: Graph of Raw Data from Logger Pro7
7
Slopes of lines that have values closest to the average slope value for each concentration of sodium chloride is shown in boxes.
Seung Soo (Jason) Lee 002213-065
Average Rate of Reaction / ΔAbss-1
Effect of Sodium Chloride Concentration on the Rate of Reaction of Amylase 0.00185
0.0016 y = -3E-05x + 0.0018 R² = 0.914 0.00135
0.0011 0
5
10
15
Sodium Chloride Concentration / % Figure 5: Graph of Average Rates of Reaction against Concentration of Sodium Chloride8 9 8 9
Vertical error bars represent standard deviation for triplicate trials. Though they are difficult to discern, horizontal error bars represent the absolute uncertainty of sodium chloride concentration.
20
Seung Soo (Jason) Lee 002213-065
Uncertainties: Standard Deviation: Rate of Reaction of Amylase / ΔAbss-1
Average / ΔAbss-1 (±Standard Deviation)
Sodium Chloride Concentration /%
Trial 1
Trial 2
Trial 3
20.0
-0.001240
-0.001280
-0.001223
0.001248 ± 0.000029
10.0
-0.001517
-0.001299
-0.001402
0.001406 ± 0.000109
5.0
-0.001812
-0.001523
-0.00123010
0.001668 ± 0.000204
1.0
-0.001836
-0.001703
-0.001714
0.001751 ± 0.000074
0.1
-0.001845
-0.001985
-0.001931
0.001920 ± 0.000071
Table 4: Standard Deviation at Different Concentrations of Sodium Chloride
Example of Standard Deviation Calculation: [Sodium Chloride Concentration] = 20%
≒ 0.000029 Same calculations were done for 10%, 5%, 1%, and 0.1% sodium chloride concentrations.
10
This value was neglected in data processing because it was considered as an outlier.
Seung Soo (Jason) Lee 002213-065
Uncertainty due to dilution of glucose solution: *Uncertainty due to 10cm3 pipette = ±0.02 cm3 Concentration of Glucose / %
20.000
Volume of sodium chloride solution added / cm3 – 3
Uncertainties Volume of distilled Total percentage 3 water added / cm error for concentration of glucose / % – – 3
Absolute uncertainty for concentration of glucose / % –
10.000
5.00 ± 0.02cm = 5.00 ± 0.4%
5.00 ± 0.02cm = 5.00 ± 0.4%
±0.80
0.008
5.000
5.00 ± 0.02cm3 = 5.00 ± 0.4%
5.00 ± 0.02cm3 = 5.00 ± 0.4%
±0.80
0.004
1.000
2.00 ± 0.02cm3 = 2.00 ± 1%
8.00 ± 0.02cm3 = 8.00 ± 0.25%
±1.25
0.013
0.100
1.00 ± 0.02cm3 = 1.00 ± 2%
9.00 ± 0.02cm3 = 9.00 ± 0.22%
±2.22
0.011
Table 5: Uncertainty for Concentration of Glucose Solution
Sodium Chloride Concentration (±Uncertainty) / %
Average Rate of Reaction (±Standard Deviation) / ΔAbss-1
20.000
0.001248 ± 0.000029
10.000 ± 0.008
0.001406 ± 0.000109
5.000 ± 0.004
0.001668 ± 0.000204
1.000 ± 0.013
0.001751 ± 0.000074
0.100 ± 0.011
0.001920 ± 0.000071
Table 6: Combined Uncertainties for Independent & Dependent Variables
Seung Soo (Jason) Lee 002213-065
Conclusions: The hypothesis was supported by the results to the extent that an increase in sodium chloride concentration decreased the rate of reaction of enzyme amylase. However, the decrease in the rate of reaction was not exponential; rather, the relationship between NaCl concentration and average rate of reaction was pretty linear. As sodium chloride concentration increased, the average rate of reaction decreased at a fairly constant rate. Furthermore, once extrapolated, the graph in figure 5 demonstrates that the rate of reaction will be 0 when the sodium chloride concentration is at 60%. From this data, one could conclude that the enzyme amylase will completely cease to catalyze reactions at NaCl concentration of 60%.
Average Rate of Reaction / ΔAbss-1
In the hypothesis, it was stated that a slight presence of sodium chloride will not affect the rate of reaction of amylase significantly, but as the concentration of sodium chloride increases, the enzyme will undergo a rapid decrease in its rate of reaction. This is due to the fact that, as more sodium chloride ions are present in amylase, the ions associate with oppositely charged groups in the enzyme protein, increasing protein hydration and denaturing the enzyme.11 Contrary to the hypothesis, where the rate of reaction was predicted to undergo a slight decrease up until a certain concentration of sodium chloride, then a rapid decrease as the concentration is at a level high enough to denature the enzyme, the graph below displays the fact that even 0.1% of sodium chloride was enough to largely decrease the rate of reaction of amylase. Although the 0.1% sodium chloride did not completely denature amylase, it was still enough to cause the greatest decrease in the rate of reaction of amylase.
Effect of Sodium Chloride Concentration on the Rate of Reaction of Amylase 0.003 0.0025 0.002 0.0015 0.001 0
5
10
15
20
Sodium Chloride Concentration / % Figure 6: Graph Demonstrating the Relationship between Sodium Chloride Concentration and Rate of Reaction, Including the Control 11
"Protein Denaturation." N.p., n.d. Web. 7 Jan 2011. <http://class.fst.ohio state.edu/FST822/lectures/Denat.htm>.
Seung Soo (Jason) Lee 002213-065
Evaluation: Overall, the results of this experiment seem fairly accurate and reliable. There are no striking outliers – except for the one value shown in table 2 – and although the standard deviations are bit sizeable for some values, they are not critical enough to negate the conclusions drawn. As the model in figure 5 represents, the relationship between sodium chloride concentration and the rate of reaction of amylase is clearly a negative correlation. On a separate note, while it is true that the best-fit line in figure 5 is a linear one, the best-fit line for the graph in figure 6 would more likely be an exponential one. Going back to one of the conclusions drawn, the relationship shown in figure 6 represents an exponential decrease because of the fact that the control is also included in the graph. The jump from 0% sodium chloride to 0.1% sodium chloride is largely significant – more significant than any of the other increases in sodium chloride concentration. Thus, such results encourage the next experiment to, perhaps, incorporate an even smaller concentration of sodium chloride. The results of this experiment support the idea that a miniscule NaCl concentration such as 0.1% was still significant enough to disrupt the electrostatic bonds within the enzyme. In order to observe the effect of NaCl concentration on the activity of enzyme more efficiently, it would be apt to utilize even more miniscule concentrations of sodium chloride. The sizeable nature of the standard deviation could be caused by the discrepancy created by human error. Although a standard was set at the beginning of the experiment, to mix the amylase and starch in the cuvette – in & out using the micropipette three times – then pressing “start” on Logger Pro, this process posed the biggest error throughout the experiment. The time taken between the moment when enzyme amylase was put into the cuvette – thus starting to interact and break down starch – and the moment when the “start” button was clicked varied, though only by little, for every trial. Furthermore, mixing the solutions in the cuvette three times – and taking up time in the process – may have been a bad idea, for that time could have been sufficient for the enzyme to do all of its work. Moreover, another problem during the procedures could have occurred with the mixing of amylase with sodium chloride. Because 15 separate micro tubes had to be filled one by one, and then mixed through the microcentrifuge one by one, some of the amylase solutions in the micro tubes had longer time to interact with sodium chloride. This could have meant longer time for the sodium chloride to denature the enzyme, thus lowering its rate of reaction. Though it’s not certain, this could have been another source of error in the experiment. Overall, however, this investigation was successful in terms of the accuracy of its results. The increased presence of sodium chloride did lower the enzyme activity of amylase, as predicted in the hypothesis, and as accepted as a scientific fact. Although improving on minor errors could strengthen the investigation, the experiment successfully produced consistent and reliable data, leading up to a solid conclusion.
Seung Soo (Jason) Lee 002213-065
Improving the Investigation: Error
Impact
Improvement
Time discrepancy between the moment amylase is inserted into the cuvette and Logger Pro reading is started
It could have allowed more time for amylase to break down starch in some trials than in others, causing differences in rate of reaction from trial to trail and increasing standard deviation
There are a few ways to improve this error. One way would be to get a help of another person, allowing him to press the “start” button on Logger Pro as soon as the amylase is mixed three times. Another method would be to simply take out the mixing process, and start the Logger Pro reading as soon as amylase is inserted into the cuvette.
Time discrepancy in the amount of time sodium chloride was allowed to interact with amylase between each trial
It allowed more time for the sodium chloride in some micro tubes to denature amylase than in other micro tubes, allowing the possibility for further decrease in the enzyme activity for amylase used in some trials compared to other trials.
All 15 micro tubes could be incubated for an allotted amount of time – around 30-40 minutes – to equalize the amount of time that sodium chloride is allowed to interact with amylase.
450μl of sodium chloride solution was inputted into the micro tube, while only 50μl of amylase solution was inputted, causing imbalance
As mentioned in the conclusion, the results demonstrate a huge decrease from no sodium chloride to 0.1% sodium chloride. This suggests that too much sodium chloride was incorporated throughout the experiment, compared to the amount of amylase. The abundance of sodium chloride could have disrupted the enzyme activity of amylase too much.
The amount of sodium chloride solution and the amount of amylase solution could be balanced, to about 250μl each used for every trial. This change could perhaps produce results that are closer to those that were hypothesized.
10 cm3 pipette used during serial dilution of sodium chloride
Decreased precision & increased range of uncertainty
Since only about 1.5ml of each sodium chloride concentration was necessary for the experiment, a micropipette could have been used to perform the serial dilution, which would have lowered the range of uncertainty.
Table 7: Ways to Improve the Investigation
Seung Soo (Jason) Lee 002213-065
Appendix: 20% Trial 1 Time (s)
20% Trial 2 Time (s)
0
Abs 614.6nm 0.325344
1
20% Trial 3 Time (s)
0
Abs 614.6nm 0.332057
0.323938
1
2
0.322158
3
10% Trial 1 Time (s)
0
Abs 614.6nm 0.313663
0.329859
1
2
0.328055
0.320197
3
4
0.319295
5
10% Trial 2 Time (s)
0
Abs 614.6nm 0.324849
0
Abs 614.6nm 0.328899
0.312331
1
0.324127
1
0.322839
2
0.310414
2
0.327022
2
0.323331
0.326182
3
0.308982
3
0.32371
3
0.323634
4
0.325192
4
0.306824
4
0.317347
4
0.318245
0.317086
5
0.321327
5
0.30595
5
0.316302
5
0.317459
6
0.316004
6
0.322082
6
0.303372
6
0.311371
6
0.315669
7
0.31459
7
0.32031
7
0.301999
7
0.312996
7
0.315408
8
0.31396
8
0.319333
8
0.301567
8
0.310745
8
0.314665
9
0.312072
9
0.317908
9
0.299804
9
0.309092
9
0.313663
10
0.310708
10
0.315929
10
0.298299
10
0.309422
10
0.311851
11
0.309642
11
0.314813
11
0.297477
11
0.309973
11
0.310488
12
0.308286
12
0.313737
12
0.296372
12
0.305913
12
0.309019
13
0.306788
13
0.312848
13
0.294631
13
0.303481
13
0.307299
14
0.305986
14
0.311556
14
0.29378
14
0.302649
14
0.305913
15
0.304786
15
0.31034
15
0.29332
15
0.301747
15
0.305404
16
0.303916
16
0.309459
16
0.292472
16
0.300774
16
0.304278
17
0.302866
17
0.308469
17
0.291767
17
0.298908
17
0.303553
18
0.302396
18
0.307774
18
0.290852
18
0.298586
18
0.302758
19
0.301495
19
0.306788
19
0.29029
19
0.297549
19
0.301639
20
0.301098
20
0.305658
20
0.289237
20
0.296479
20
0.300631
10% Trial 3 Time (s)
5% Trial 1 Time (s)
0
Abs 614.6nm 0.32863
1
5% Trial 2 Time (s)
0
Abs 614.6nm 0.339073
0.327137
1
2
0.324811
3
5% Trial 3 Time (s)
0
Abs 614.6nm 0.322687
0.333645
1
2
0.329398
0.322271
3
4
0.323028
5
1% Trial 1 Time (s)
0
Abs 614.6nm 0.346723
0
Abs 614.6nm 0.320573
0.320762
1
0.342346
1
0.320573
2
0.316787
2
0.34045
2
0.31675
0.328362
3
0.314999
3
0.341792
3
0.312109
4
0.328055
4
0.314108
4
0.339938
4
0.310892
0.320875
5
0.322536
5
0.31115
5
0.333917
5
0.30781
6
0.32178
6
0.320988
6
0.31012
6
0.332986
6
0.30635
7
0.31847
7
0.31832
7
0.31034
7
0.331864
7
0.302613
8
0.315743
8
0.316675
8
0.306642
8
0.331285
8
0.301711
9
0.314293
9
0.316302
9
0.305113
9
0.330668
9
0.299194
10
0.311814
10
0.313515
10
0.303843
10
0.329744
10
0.299266
Seung Soo (Jason) Lee 002213-065 11
0.311556
11
0.312183
11
0.3033
11
0.328515
11
0.296978
12
0.31012
12
0.310561
12
0.301567
12
0.327787
12
0.294701
13
0.308506
13
0.308359
13
0.299445
13
0.326144
13
0.292014
14
0.306605
14
0.307189
14
0.298514
14
0.325039
14
0.290817
15
0.306532
15
0.306059
15
0.299338
15
0.3239
15
0.289623
16
0.304641
16
0.304097
16
0.298192
16
0.323255
16
0.288467
17
0.303952
17
0.303807
17
0.296408
17
0.322498
17
0.287106
18
0.303264
18
0.302469
18
0.295625
18
0.321667
18
0.286131
19
0.302721
19
0.30135
19
0.294276
19
0.321365
19
0.285609
20
0.301387
20
0.300092
20
0.294063
20
0.320235
20
0.286235
1% Trial 2 Time (s)
1% Trial 3 Time (s)
0
Abs 614.6nm 0.337622
1
0.1% Trial 1 Time (s)
0
Abs 614.6nm 0.337035
0.334461
1
2
0.332212
3
0.1% Trial 2 Time (s)
0
Abs 614.6nm 0.342465
0.33485
1
2
0.33256
0.33009
3
4
0.327749
5
0.1% Trial 3 Time (s)
0
Abs 614.6nm 0.341121
0
Abs 614.6nm 0.326678
0.340529
1
0.340253
1
0.323066
2
0.337152
2
0.335317
2
0.321252
0.329898
3
0.333762
3
0.331246
3
0.318695
4
0.327864
4
0.331169
4
0.328208
4
0.316712
0.325192
5
0.325915
5
0.329475
5
0.326831
5
0.31333
6
0.323824
6
0.324241
6
0.326946
6
0.324925
6
0.311224
7
0.321365
7
0.322271
7
0.325687
7
0.322536
7
0.308872
8
0.319859
8
0.320423
8
0.323407
8
0.320047
8
0.307189
9
0.317721
9
0.317908
9
0.321554
9
0.317983
9
0.305258
10
0.315743
10
0.316302
10
0.319145
10
0.316339
10
0.302938
11
0.314331
11
0.314776
11
0.317684
11
0.314145
11
0.301423
12
0.31333
12
0.313293
12
0.316228
12
0.312737
12
0.300056
13
0.311888
13
0.311851
13
0.314702
13
0.310819
13
0.29812
14
0.310782
14
0.310488
14
0.313218
14
0.309679
14
0.296194
15
0.309422
15
0.308579
15
0.311298
15
0.308469
15
0.294595
16
0.308213
16
0.307372
16
0.31012
16
0.306642
16
0.29279
17
0.30635
17
0.306532
17
0.308909
17
0.304822
17
0.291696
18
0.304931
18
0.304822
18
0.30708
18
0.303409
18
0.290044
19
0.303011
19
0.303988
19
0.306205
19
0.302252
19
0.288992
20
0.302541
20
0.302685
20
0.30504
20
0.300056
20
0.287315
Seung Soo (Jason) Lee 002213-065 Maximum Absorbance Wavelength (nm) 400
Abs 0.140983
Wavelength (nm) 523.68
403.5
0.143298
407
Control (No NaCl) Abs
Time (s)
0.269512
Wavelength (nm) 629.44
0.506694
0
Abs 614.4nm 0.356633
526.6
0.276394
632.4
0.505749
1
0.351468
0.144298
529.52
0.284868
635.36
0.502435
2
0.352439
410.5
0.143858
532.44
0.293012
638.32
0.499854
3
0.345206
414
0.144867
535.36
0.303899
641.28
0.497412
4
0.341121
417.5
0.145449
538.28
0.313062
644.24
0.493168
5
0.34116
421
0.144269
541.2
0.322568
647.2
0.492372
6
0.337113
424.5
0.15575
544.12
0.332483
650.16
0.488485
7
0.337543
428
0.16144
547.04
0.34552
653.12
0.482216
8
0.329129
431.5
0.168923
549.96
0.356925
656.08
0.480191
9
0.327175
435
0.171384
552.88
0.367604
659.04
0.474983
10
0.323634
438.5
0.177023
555.8
0.378477
662
0.47441
11
0.321214
442
0.18106
558.72
0.389967
664.96
0.469595
12
0.318058
445.5
0.185164
561.64
0.402134
668
0.464303
13
0.314479
449
0.184681
564.56
0.413991
671
0.461757
14
0.312368
452.5
0.164762
567.48
0.424809
674
0.455878
15
0.311999
456
0.184657
570.4
0.435809
677
0.451873
16
0.309202
459.5
0.184207
573.32
0.446248
680
0.448952
17
0.306168
463
0.183719
576.24
0.456774
683
0.445418
18
0.304133
466.5
0.19358
579.16
0.46569
686
0.438446
19
0.303011
470
0.198735
582.08
0.47381
689
0.43383
20
0.301639
473.5
0.205575
585
0.48009
692
0.428318
477
0.209701
588
0.487579
695
0.425969
480.5 484
0.213411 0.216026
590.96
0.490925
698
0.42161
593.92
0.492981
701
0.416259
487.5
0.218792
596.88
0.501117
704
0.411412
491
0.221762
599.84
0.504044
494.5
0.224311
602.8
0.507599
498
0.227361
605.76
0.511082
501.5
0.232694
608.72
0.510439
505
0.235745
611.68
0.51301
508.5
0.240517
614.64
0.514248
512
0.245664
617.6
0.512766
514.92
0.253401
620.56
0.510628
517.84
0.258245
623.52
0.509945
520.76
0.264801
626.48
0.508682
Abs