MUJOMBI TALENT CHIPO

R143547B

MBChB BIOCHEMISTRY PART 1

EXPERIMENT 1: VOLUMETRIC
DETERMINATION OF A
MIXTURE OF NaOH AND Na2CO3
BY ACID-BASE TITRATION.

INTRODUCTION
An acid–base titration is the determination of the concentration of an acid/base by exactly
neutralizing the acid/base with an acid/base of known concentration. This allows for quantitative
analysis of the concentration of an unknown acid/base solution. It makes use of the neutralization
reaction that occurs between acids and bases. The analyte (titrand) is the solution with an
unknown molarity. The reagent (titrant) is the solution with a known molarity that will react with
the analyte. The analyte is prepared by dissolving the substance being studied into a solution.
The solution is usually placed in a flask for titration. A small amount of indicator is then added
into the flask along with the analyte. The reagent is usually placed in a burette and slowly added
to the analyte and indicator mixture. The amount of reagent used is recorded when the indicator
causes a change in the color of the solution.  A useful indicator has a strong color that changes
quickly near its pKa. These traits are desirable so only a small amount of an indicator is needed.
If a large amount of indicator is used, the indicator will affect the final pH, lowering the accuracy
of the experiment. The indicator should also have a pKa value near the pH of the titration's
endpoint. Choosing an indicator with a pKa near the endpoint's pH will also reduce error because
the color change occurs sharply during the endpoint where the pH spikes, giving a more precise
endpoint. (Ramsden, 2000)

Sodium hydroxide - both solid and dissolved - easily reacts with atmospheric carbon dioxide.
That means it is usually contaminated with disodium carbonate Na2CO3. (Harris, 2004). It is not
a problem to determine sum of hydroxide and carbonates concentration by titration with a strong
acid. It is possible to determine both sodium hydroxide and carbonate concentration in one
titration; this method is very fast, simple and accurate enough.

Bio-chemical significance of acid-base titration

Acid–base homeostasis is the part of human homeostasis concerning the proper balance between
acids and bases, also called body pH. Almost every biological process is pH dependent; a small
change in pH produces a large change in the rate of the process. This is true not only for the
many reactions in which the H+ ion is a direct participant, but also for those in which there is no
apparent role for H+ ions. The enzymes that catalyze cellular reactions, and many of the
molecules on which they act, contain ionizable groups with characteristic pKa values. The
protonated amino and carboxyl groups of amino acids and the phosphate groups of nucleotides,
for example, function as weak acids; their ionic state depends on the pH of the surrounding
medium. As we noted above, ionic interactions are among the forces that stabilize a protein
molecule and allow an enzyme to recognize and bind its substrate. Cells and organisms maintain
a specific and constant cytosolic pH, keeping biomolecules in their optimal ionic state, usually
near pH 7. In multicellular organisms, the pH of extracellular fluids is also tightly regulated.
Constancy of pH is achieved primarily by biological buffers: mixtures of weak acids and their
conjugate bases. (Nelson and Cox, 2005) Since many metabolic reactions are accompanied by
the release or uptake of protons, most intracellular reactions are buffered. Oxidative metabolism
produces CO2, the anhydride of carbonic acid, which if not buffered would produce severe
acidosis. Maintenance of a constant pH involves buffering by phosphate, bicarbonate, and
proteins, which accept or release protons to resist a change in pH (Murray et al, 2003).
Bicarbonate, an intermediate in this reaction, is an important buffer salt in the cell and body. The
CO2–bicarbonate buffer is a little different from buffers using the usual kind of acids and bases, but it is
extremely important in maintaining the acid–base balance of the blood (Gilbert, 2000). Therefore, the
objective of this practical is:
 To determine the concentration of NaOH and Na2CO3 in the aliquot solution

Materials
“As per biochemistry practical schedule MBChB/BDS 2014-2015”

Method
The researcher titrated the aliquot with HCl using phenolphthalein and methyl orange as
indicators in the first and second phase respectively.

Results
Table 1: Summary of results from acid-base titrations

PROCEDURE TITRE

V1 (ml) V2 (ml)

1ST procedure 20.70 4.10

2ND procedure 20.60 4.20

3RD procedure 20.60 4.20

Mean 20.63 4.17

Mean = Volume in 1st + Volume in 2nd + Volume in 3rd procedure
3

Discussion
Solution contains three bases: OH-,HCO3-,CO32-. According to (Ramsden, 2004), the stronger the
base, the easier it reacts with acid. Of the three bases present, NaOH was the strongest, so it was
neutralized first. This was then followed by the carbonate ion. When all CO32- was converted to
HCO3-, pH of the solution dropped to approximately 8.31.Solution was decolorized from pink
after 20.63ml of HCl were added. This change of color was due to phenolphthalein indicator
which is known to change color at a pH of 8.2 (Chang.2005). After this first titration, a solution
of HCO3- ions remained. That means after adding V1 (20.63) of the titrant we have titrated all of
sodium hydroxide and some of the sodium carbonate. The reactions that occurred in phase 1
were:

NaOH + HCl → NaCl + H2O

Na2CO3 + HCl → NaCl + NaHCO3

Since HCl is reacting with two bases in phase 1,a larger volume of it will be required than in
phase 2.

When the neutralization of NaOH is complete, and neutralization of Na2CO3 is partially

complete, the remaining solution now contains only one base,HCO3-. Once it gets protonated, we
will be left with solution of carbonic acid, with pH around 4.0. This is close to the pH at which
methyl orange starts to change color (4.4). Reaction that took place during this second stage of
titration was:

NaHCO3+ HCl → NaCl + H2O + CO2

This means that after adding V2(4.17),all the remaining NaHCO3 will have been neutralized.
Less volume of acid is required here as we are neutralizing only one base

Calculations
ai) Volume of HCl used to neutralize NaOH = 20.63ml - 4.17ml
16.46ml

aii) Volume of HCl used to neutralize all NaHCO3 = 4.17 x 2

8.34

b) # of moles HCl used in to neutralize NaOH = CV

= 0.1mol/dm3 x 16.46
1000
= 0.001646mols

Therefore #mols of NaOH = 0.001646mols

Mass of NaOH = 0.001646 x 40

=0.06584g

[NaOH] = mass = 0.06584g = 2.6336g/l

volume 25/1000

#moles HCl /used to neutralize NaHCO3 = CV

= 0.1 mol/dm3 x 4.17
1000
= 0.000417mols

Therefore #mols Na2CO3 = 0.000417mols

Mass of Na2CO3 = 0.000417 x 106

= 0.044202g

[Na2CO3] = 0.044202 = 1.76808g/l

25/1000

The concentrations of both Na2CO3 and NaOH (as calculated above), were in the range that had
been specified as per Biochemistry Practical schedule,2014-2015.

Answers to questions:
ai) proton donor
 ii) proton acceptor

bi) pH is the negative logarithm to base 10 of the hydrogen ion concentration

ii) pH = -log[6.5x10-5]
= 4.19

c) A buffer is a solution that has both the acidic and basic forms of a weak acid and it resists
changes in pH when a little acid/alkali is added.

Mechanism of the bicarbonate buffer

CO2 + H2O H2CO3 + H+

H2CO3 H+ + HCO3-

H2CO3 dissociates partially according to equation 2 above under physiological pH. On addition
of H+ ions, the HCO3- ion reacts to produce H2CO3 hone cancelling the acidifying effect the added
H+ ions. Upon addition of OH-, the H+ ions produced from the partial dissociation of H2CO3
react with to produce water which is neutral.

d) Indicators change color over a wide range of pH thus reducing accuracy

-since indicators are weak acids, they sometimes react with the bases thus reducing accuracy also

Conclusion
In the phase of this evidence therefore, the researcher concluded that1.76808g/l of Na2CO3 and
2.6336g/l of NaOH reacted with the 0.1mol/dm3 HCl. They also concluded that alkali/acid
solutions can be determined volumetrically by acid/base titrations.

References
Chang, H. (2005) Chemistry. 2nd edition. (Macmillan Publishing Co.: New York), pg 30-33
Gilbert, H.F. (2000). Basic concepts of Biochemistry, A student’s survival guide. 2nd Edition
(McGraw Hill: USA), pg255

Harris, D. (2004) Exploring chemical analysis. 3rd Edition. (W.H Freeman: New York)

Murray, R. K., Granner, D. K., Mayes, P. A., Rodwell, V.W. (2003). Harper’s illustrated
biochemistry. 26th edition. (McGraw Hill Companies: Toronto), pg22-23

Nelson, D.L., Cox, M.M. (2005) Lehninger, Principles of Biochemistry. 4th Edition (W.H
Freeman: New York), pg 65-66

Ramsden, E.N. (2000). A 'Level Chemistry. 4th Edition. (Nelson Thornes: USA)