CHM 224: Physical Chemistry Laboratory

Experiment 7: Acidic and basic dissociation constants of glycine and its

isoelectronic point

Theory:
The isoelectric point or isoelectronic point (commonly denoted as pI) is the pH at which a molecule or
surface carries no net electrical charge. It is at this pH that the molecule will not show any motion in an
electric field. In order to have a sharp isoelectric point, a molecule (or surface) must be amphoteric,
meaning it must have both an acidic and basic functional groups. Proteins and amino acids are common
molecules that meet this requirement.
For an amino acid with only one amine and one carboxyl group, the pI can be calculated from the pK a
values of this molecule.
H H

H H + H+ pKa1
HOOC -
OOC
NH3+ NH3+
H H

- H H + H+ pKa2
OOC -
OOC
NH3+ NH2

pKa1 + pKa2
pI =
2

Chemicals:
Glycine, sodium hydroxide

Glassware & Equipment required:

pH meter, burette, beaker (100 mL), beaker (500 mL)

Procedure:

(A) Standardization of the pH meter

1. Switch on the pH meter and allow the instrument to warm up for 10 minutes.
2. Set the temperature control to room temperature.
 3. Rinse the electrode with distilled water and pat gently using tissue paper. (Do not rub the
electrode with tissue paper)
4. Insert the electrode assembly into the provided buffer solution (pH 4)
5. Set the selector switch of the instrument to read pH. If it does not show a value of 4 reset the
value to 4 using the calibration knob.
6. Remove the electrode assembly, rinse in distilled water, pat dry and insert in provided buffer
solution (pH 9.2) Calibrate again.
7. Repeat this with the provided buffer (pH 7)

(B) Determination of pI of glycine

1. Prepare a 0.1 M sodium hydroxide
solution (100 ml) in a 250 ml reagent
bottle.
2. Take 25 ml of the supplied 0.1 M
glycine hydrochloride solution in a clean
dry 100 mL beaker and measure the pH.
3. Add sodium hydroxide to the glycine
hydrochloride solution in the sequence
provided below:
(i) 3 mL till pH reaches 3.5
(ii) 0.5 mL till pH reaches 8.5
(iii) 3 mL till pH reaches 12
4. Record the reading as the amount of
sodium hydroxide added versus pH.
5. Plot a graph with volume of NaOH on
the x-axis and pH on the y-axis.
6. The two almost horizontal parts of the
graph gives the values of pK a1 and pK a2
for glycine. Use mid-points of these regions to get the values.
7. The average of these values (pK a1 and pK a2 ) gives the pI of glycine.

Note down all the observations in your lab notebook. Clean all your glassware and hand over to the
lab attendant.
 CHM 224: Physical Chemistry Laboratory

Experiment 8: Kinetics of saponification of ethyl acetate using conductometry

Theory:
The alkaline hydrolysis of an ester is called saponification. Ethyl acetate on alkaline hydrolysis
produces ethanol and acetate.
CH3COOC2H5 + OH- CH3COO - + C2H5OH
t=0 a a 0 0
t=t a-x a-x x x

The overall reaction is kinetically of second order, being first order with respect to each of
reactants, the ester and hydroxyl ions (OH-). The rate of the overall reaction maybe expressed
according to (1)
Rate = -d[ester]/dt = k[ester][OH-] ------------------- (1)
Where k is the rate constant in mole-1.litre.second-1 and [ ]s stand for concentrations in moles/L.


= . ( )2 --------------------- (2)

Integration of equation (2), for x = 0, when, t = 0 yield

1
=
. ---------------------- (3)

The progress of the reaction can be monitored by measuring the electrolytic conductance of the
reaction mixture. If C 0 , C t and C be the conductances of the reaction mixture at time t = 0, t and
at completion of the reaction (infinite time, t = ), then, equation (3) is transformed to:
(0 )
( )
= . . ---------------------- (4)

A plot of [(C 0 C t ) / (C t - C )] versus t will be a straight line passing through the origin and
possessing a positive slope = .

Chemicals:
Ethyl acetate, Potassium Hydrogen phthalate, sodium hydroxide, phenolphthalein

Glassware & Equipment required:

Conductivity meter, volumetric flask (100 ml), Reagent Bottle (250 ml), Pipette (10 ml), Burette,
beaker
Procedure:
1. Prepare 100 mL 0.05 M potassium hydrogen phthalate solution by accurate weighing in
a volumetric flask.
2. Prepare 100 mL of 0.05 M NaOH solution in a 250 mL reagent bottle. Use conductivity
water for making this solution.
3. Fill up the burette with sodium hydroxide solution.
4. Pipette out 10 ml of potassium hydrogen phthalate solution in a clean dry 100 ml conical
flask.
5. Add two drops of phenolphthalein and titrate against sodium hydroxide till a faint pink
color is obtained. Note down the burette reading.
6. Repeat the titration and calculate the strength of sodium hydroxide solution.
7. Pipette out 10 ml of supplied acetic acid solution (0.07N) in a clean dry 100 ml conical
flask.
8. Add two drops of phenolphthalein and titrate against the standard sodium hydroxide
solution till a faint pink color is obtained. Note down the burette reading.
9. Repeat the titration and calculate the strength of acetic acid solution.

Preparation of solutions for conductivity measurement:

10. Prepare 100 mL of exact 0.02 N sodium hydroxide solution by proper dilution in a 100
mL volumetric flask (use conductivity water).
11. Take 25 mL of this solution is a clean dry 100 mL beaker and measure the conductance
(C 0 ).
12. Prepare 100 mL of exact 0.02 N acetic acid from the supplied 0.07 N acetic acid by
proper dilution in a 100 mL volumetric flask (use conductivity water).
13. Prepare an exact 0.01 N sodium acetate CH3COO-Na+ by mixing equal volumes of 0.02
N acetic acid and 0.02 N sodium hydroxide. (Mix 20 mL of 0.02 N acetic acid and 0.02
N sodium hydroxide in a 100 mL beaker, stir well with a glass rod). Measure the
conductance (C ).
14. Prepare 100 mL of exact 0.02 N ethyl acetate solution from the 0.05 N ethyl acetate
provided to you.
15. In a dry 100 mL beaker, take 20 mL of 0.02 N sodium hydroxide and 20 mL of 0.02 N
ethyl acetate solution (use pipette) and note the time of half discharge.
16. Measure the conductance (C t ) of the reaction mixture at different time intervals,
approximately 1, 2, 3, 5, 7, 10, 12, 15, 25, 40, . Till the conductance remains
practically unchanged with time. Record the conductance vs time data in a tabular form.
Record the temperature of the experiment.
17. Plot (C 0 -C t )/(C t -C ) against t to obtain rate constant, k from the slope.

Note down all the observations in your lab notebook. Clean all your glassware and
hand over to the lab attendant.
 CHM 224: Physical Chemistry Laboratory

Experiment 9: Potentiometric Titration of a standard solution of AgNO 3

against mixture of halides Determination of concentration of two salts in a
given mixture

Theory:
The determination of the equivalence point of titrations based on potential measurements is
called potentiometric titrations. The electrode potential of an electrode depends upon the
concentration of its ions in the solution. Hence the potential of an indicator electrode goes on
changing with respect to a standard reference electrode by the change of concentration of the
ions during the titration. The equivalence point is indicated by a fairly large change in potential.
When two different redox electrodes are electrochemically connected, an electrochemical cell is
formed which is conventionally represented as
(-) (+)
(Left hand electrode) (E L ) || Right hand electrode) (E R )
Oxidation occurs Reduction occurs
When the Ag+|Ag(s) redox electrode is coupled with a saturated calomel electrode (SCE), KCl
(satd). Hg 2 Cl 2 (s).Hg(l), the following electrochemical cell is produced.
(-) (+)
(Pt) Hg(l) | Hg 2 Cl 2 (s) |KCl (aq. Saturated) || Ag+ (aq) | Ag (s)
(E L ) (E R )
Where, the symbol, II, stands for agar 2(M) NH 4 NO 3 salt-bridge, which eliminates the liquid
junction potential. The half cell reactions at the electrodes are:
L. H. Electrode: 2 Hg (I) + 2Cl- (aq) Hg 2 Cl 2 (s) +2e-
R. H. Electrode: Ag+ (aq) + e- Ag (s)
The overall reaction is:
2Hg (l) + 2 Cl- (aq) + 2 Ag+ (aq) Hg 2 Cl 2 (s) + 2Ag(s)
And e.m.f of the cell is given by:
E cell = E R E L = E Ag+|Ag - E SCE
= E0 Ag+|Ag + 0.059 log [a Ag+ |a Ag(s) ] E SCE (at 250C)
Ag(s) being in the standard state, its activity will be unity. For a dilute solution activity (a) of
Ag+ ion may be replaced by the numerical value of its concentration [Ag+]. Thus,
E cell = E0 Ag+|Ag + 0.059 log[Ag+] - E SCE (1)
Since E0 Ag+|Ag and E SCE are fixed,E cell depends on [Ag+].
As AgNO 3 is added to the solution containing a mixture of halides (KCl and KI), the initial
additions result in the formation of AgI exclusively, owing to a much lower solubility product
compared to AgCl followed by the formation of AgCl.

Chemicals:
Silver Nitrate, potassium chloride, potassium iodide.

Glassware & Equipment required:

Volumetric Flask (100 mL x2), beaker (100 mL x2), NH 4 NO 3 salt bridge, pipette, burette,
saturated calomel electrode, fully saturated solution of KCl, silver electrode, potentiometer,
magnetic stirring bar.

Procedure:
1. Prepare 100 mL of a standard 0.1 M AgNO 3 solution in conductivity water by accurate
weighing (use volumetric flask)
2. Take 10 mL of the prepared AgNO 3 solution (use pipette) in a 100 mL beaker and dip the
silver electrode in this solution. Add sufficient amount of conductivity water (use pipette)
so that the electrode dips properly. This constitutes experimental electrode (Ag+|Ag(s)).
3. Set up the experimental cell by connecting the saturated calomel electrode (SCE) dipped
in a fully saturated solution of KCl (supplied) and the experimental electrode through the
agar-NH 4 NO 3 salt bridge (Supplied).
4. Now connect the experimental cell with the potentiometer. (Ensure that you insert the
jacks in the proper sockets).
5. Calculate the emf of the cell at this point of the time using equation 1. (E0 Ag+|Ag =0.799 V
and E SCE =0.2415 V).
6. Now using the set zero knob on the potentiometer set the emf at the value calculated.
7. Now discard the AgNO 3 solution and take 20 mL of supplied KCl and KI solution
mixture in a 100 mL beaker. Dip the silver electrode in this solution. Add sufficient
amount of conductivity water so that electrodes dip in properly (use pipette).
8. Insert a magnetic stir bar into the KCl and KI solution mixture for homogenous mixing
(be careful, the magnetic stir bar should not hit the silver electrode).
9. Now add AgNO 3 from burette and note down the emf of the cell E cell everytime in the
sequence provided below:
(i) 2 drops at a time for the first 15 additions
(ii) 1 drop at a time for the next 20 additions
 (iii) 2 drops at a time for the next 10 additions
(iv) 1 drop at a time for the next 20 additions
10. By this time the emf value would have reached constancy. If not add a few more drops.
11. Record the number of drops of AgNO 3 and the E cell values in tabular form.
12. Plot the observed E cell values against number of drops of AgNO 3 solution added.
13. Determine the equivalence points and the E cell values at the equivalence points from the
graph.
14. Hence calculate the molarities of KCl and KI supplied to you.

Note down all the observations in your lab notebook. Clean all your glassware and hand
over to the lab attendant.