Completing lab data

1.
-

2.
-

Acids, bases and pH

Concentration of H3O in each solution as HCl and NaOH is added
Draw a summary table
Explain the changing pH values you have recorded, as you added
either HCl or NaOH to water
Observation
In the H2O with added HCl solution, the pH drastically decreased to 1.5pH
with the first .5mL of HCl drop in 200mL of water. With the second .5mL
drop, there were only .3 differences in the pH value and then it became .2
and .1 decrease on the third and forth .5mL drops. Changes in H3O+
concentration were significantly large during the first two .5mL drops of
HCl. Smaller changes were recorded after. This is due to most of the water
molecules have reached its

pH buffers
Equation for the reaction of the two compounds involved.
Plot your data graphically
Derive the dissociation constant (pKa) of the acid under test
Compare (and explain) the results of this experiment to the
results you got when adding HCl to water

3. Amino acids
- plot the data from this experiment graphically
- determine the two pKa values for glycine hydrochloride
(i)

(ii)
(iii)
(iv)

explain why there are two pKa values for the amino acid glycine
describe the order of proton dissociations from the molecule
determine these two values from your graph and your
understanding of the Henderson-Hasselbalch equation
show the various dissociation states of glycine as the pH changes
across the titration curve

Title: pH titration of a dibasic acid.

Aims:

To carry out a potentiometric determination of glycine hydrochloride and to

estimate pKa values for the -amino acid.
To use the values obtained to calculate the theoretical isoelectric point of glycine.
To assay the content of pure compound (C2H6NO2Cl)2 in the sample.
Able to be perform simple calculations involving the concepts of mass, volume,
concentration and the mole. To provide the basis for further investigations of acidbase reactions.

Introduction:

-amino acid are difunctional which contain both carboxylic acid group and amine
group.

Peptide and proteins are linked up by peptide bonds between amino acids.
Amino acid are amphoteric because it may act as either acid or base.
In solution, they act as internal ions or zwitterions.
Amino acid are very polar. Hence, very soluble I water and form crystalline solids.
Carboxylate group becomes more deprotonated when pH fall due to NH3+.
NH3+ loses its proton as pH rises due to COO-.
At certain pH value specific for each amino acid will be no net charge.
This pH is known as isoelectric point whereby the amino acid will not move in an
electrical field.
Example: Glycine, the isoelectric point is the average of the two pKa values.
pKa values for readily water soluble compounds, with moderately strong acidic and
basic functional group can be obtained by potentiometric titration which is based
on Henderson-Hasselbalch Equation that relates the fraction of ionized to unionized
form for any functional group to its pKa value and the pH of the solution.

Methods
1

1g of glycine hydrochloride is accurately weighed using a Balance B by difference

approximately 1g of glycine hydrochloride.

Weight of sample transferred is recodered.

Sample is dissolved with about 50mL of distilled water in a suitable beaker.

Use the electronic magnetic stirrer device. Place a magnetic bar into the 50mL
solution, and turn on the stirrer so that the bar turns at a moderate speed.
magnetic bar is checked so it does not collide with the electrode.

Record the initial pH

Titration stand is assembled, and pH electrode is submerged into the solution and
the burette is positioned to drop solution into the beaker.

The solution is titrated with standardized 0.5160M sodium hydroxide solution using
the potentiometric apparatus.

Ensured small aliquots of titrant are added in the region of the inflection points
and continued until both groups are titrated (approximately 40mL)

1.0mL of NaOH was added from the burette. This step was contibued until reached
2.0mL before end point.

10 When it approaches the end point which is between 17.0 mL and 19.0mL, the
procedure continued by adding the base in 0.2mL aliquots.
11 The pH value was noted after each addition until a sharp rise in pH was observed.
12 After the first inflection point was obtained, the pH was recorded subsequently
with every 1.0mL of titrant.

13 0.1mL of titrant was added continuously when the volume of the titrant approaches
the twice amount of the titrant volume gained at the first inflection point.
14 This was continued until the nearest whole number was obtained.
15 The exact molarity of NaOH(M) is recorded.
16 Results are recorded in the tables provided.
17 The titration was repeated for sample 2.

Discussion:
Amino acids such as Glycine are more complicated than simple weak acids since
amino acids have at least 2 ionizing groups.
Glycine has both a carboxylic acid and an amino group that can ionize, therefore
they are dibasic.
If we dissolve the free base of Glycine in pure water (ie neutral pH), it will ionize
by protonating itself.
The equilibrium is far to the right so most of the Glycine is in the charged form
called the Zwitterion and Glycine is still neutral because the + charge is netualized
by the - charge.
Glycine is always in the Zwitterion form at neutral pH.
Now if we put Glycine at an acid pH where it is fully protonated.
Glycine is neutral at pH 6; it has no net charge here.
The deviation of the pKa1 and pKa2 values for both samples will also cause the
values of the theoretical isoelectric point of both samples to be deviate from the
accurate theoretical isoelectric point.
Conclusions:
Carrying out potentiometric titration of glycine hydrochloride can estimate the inflection
points and pKa values of glycine hydrochloride.
Each sample has two inflection points and two pKa values.
From the graph of pH against volume of NaOH added, we can estimate the two inflection
points.
The theoretical isoelectric point of glycine hydrochloride can be obtained from both pKa
values from the sample which is the average value. Each of the samples has the same
value of theoretical isoelectric point.
The weight of the pure compound in the sample can also be calculated based on the first
inflection point. Finally, the weight of the pure compound in the sample is expressed as
percentage purity (%w/w).

Reference:
http://www.unm.edu/~rrobergs/titration.htm
http://www.bio.mtu.edu/campbell/aminot1.htm
Laboratory Guide of MPH115/CHM113;University of Sunderland; Year 2006
Donald Cairns; Essentials of Pharmaceutical Chemistry; Pharmaceutical Press;Year 2000;
page 16-17
Statement of Authenticity
I declare that the report above is completed all by my own effort and there are some of
the information taken from books and websites. However, all of the books and websites
are stated at section Reference above.

Signature:
Date:

Graph and calculation

Data 1

Volume
(mL)

pH

Volume
(mL)

pH

Volume
(mL)

pH

0
1

1.58
1.66

18.5
18.6

4.13
4.2

26
27

9.59
9.7

1.81

18.7

4.45

28

9.79

1.95

18.8

5.32

29

9.9

2.13

18.9

6.1

30

9.98

2.3

19

31

10.08

11

2.48

19.1

7.57

32

10.2

13

2.69

19.2

7.83

33

10.31

15

2.91

19.3

7.97

34

10.48

17

3.32

19.4

8.05

35

10.63

17.2

3.36

19.5

8.24

36

10.83

17.4

3.42

19.6

8.3

36.5

10.97

17.6

3.52

19.7

8.4

37

11.2

17.7

3.58

19.8

8.46

37.2

11.26

17.8

3.6

19.9

8.48

37.4

11.33

17.9

3.63

20

8.53

37.6

11.41

18

3.75

21

8.88

37.8

11.63

18.1

3.79

22

9.06

38

11.73

18.2

3.84

23

9.22

38.5

11.87

18.3

3.9

24

9.37

39

12

18.4

25

9.49

39.5

12.06

40

12.14

Second Infection
point
= 37.8 mL
First Infection point = 18.9 mL

Data 2

Volume
(mL)

pH

Volume
(mL)

pH

Volume
(mL)

pH

0.1
1.0

1.62
1.69

16.8
17.0

4.15
4.48

29.0
30.0

10.13
10.24

2.0

1.78

17.1

6.48

31.0

10.38

3.0

1.87

17.2

7.22

32.0

10.55

4.0

1.97

17.3

7.72

33.0

10.77

5.0

2.05

17.4

8.02

33.5

10.92

6.0

2.15

17.5

8.11

34.0

11.11

7.0

2.25

17.6

8.22

34.1

11.15

8.0

2.34

17.7

8.31

34.2

11.19

9.0

2.43

17.8

8.40

34.3

11.23

10.0

2.53

17.9

8.44

34.4

11.27

11.0

2.64

18.0

8.54

34.5

11.31

12.0

2.75

18.5

8.69

34.6

11.35

13.0

2.88

19.0

8.88

34.7

11.40

14.0

3.03

20.0

9.08

34.8

11.47

15.0

3.23

21.0

9.29

34.9

11.51

15.5

3.38

22.0

9.38

35.0

11.55

16.0

3.57

23.0

9.50

36.0

11.88

16.2

3.68

24.0

9.62

37.0

12.10

16.4

3.77

25.0

9.72

38.0

12.19

16.6

3.94

26.0

9.82

39.0

12.19

27.0

9.92

40.0

12.22

28.0

10.03

Second Infection point

= 34.0 mL

First Infection point = 17.0 mL