Laboratory Manual for Practical Exercises Acids and bases

Acids and bases

Contents
Acids and bases ........................................................................................................................1
Measurement and calculation of pH ........................................................................................2
Exercise: pH determination by acid-base indicators ...................................................................7
Exercise: pH determination by natural indicators – antocyanidins ...............................................8
Exercise: pH determination of body and natural fluids ...............................................................9
Exercise: Measuring pH by pH-meter .................................................................................... 10
Buffers .................................................................................................................................. 11
Exercise: Preparation of acetate buffers ................................................................................. 12
Exercise: Effect of acids and bases on pH of buffers, Buffering capacity .................................... 13
Control questions: ............................................................................................................... 14

Reactions between acids and bases belong among the most common chemical reactions in inorganic
chemistry. General concept of acids and bases was introduced by Brönsted. The basic postulate is
definition of acid as a substance capable of giving up a proton and base as a substance binding the
proton. Acid is a donor of proton, base is an acceptor of proton. In the acid-base equilibrium the acid
and base form a conjugate acid-base pair. Both partners have to be in solution at the same time, e.g.
HCl + NH3 → Cl- + NH4 +
According to the Brönsted's theory, some of ions (cationic or anionic) may be acids and bases, as
well. Acids and bases have the common name - protolytes and we used to call reactions between them
as protolytic.
Individual reaction represents autoprotolysis. It is a reaction where two molecules of the same substance
in function of Brönsted's acid and base transfer proton to each other. As an example we can use:
H2O + H2O ↔ H3O+ + OH-
NH3 + NH3 ↔ NH4+ + NH2-
H2SO4 + H2SO4 ↔ H3SO4+ + HSO4-
Therefore, e.g. in case of autoprotolysis of water, ionization constant of an autoprotolytic reaction has
form:
[H3O+][OH-]
Kc=
[H2O]2
Ionization of a solvent is low, therefore we can consider concentration of not ionized molecules as
constant. Thus, it is possible to establish new constant, ion product, or more precisely autoprotolytic
constant of solvent Kaut, which for a water has a form:
Kaut (KW) = Kc.[H2O]2 = [H3O+][OH-]

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Table: Autoprotolytic constants of solvents at 18 to 25 ºC

Solvent Kaut Solvent Kaut
Ethanol 7.9x10-20 Sulphuric acid 2.5x10-4
Formic acid 6.3x10-7 Hydrogen peroxide 1x10-13
Acetic acid 3.2x10-15 water 1x10-14

Measurement and calculation of pH

An activity of hydrogen ions in solution is expressed by hydrogen exponent pH. Term hydrogen
exponent introduced Sörensen in order to simplify indication of hydrogen ions concetration. Determing
(measurement) of pH belongs to fundamental tasks in chemical laboratories. Several methods are used.
Those simple include colorimetric methods using various acid-base indicators. As acid-base indicators
are used some of organic compounds, that are sensitive on change of hydrogen ions concentration. They
also change their colour in the dependence on pH of medium, in which they are dissolved. They are
weak acids, eventually bases and their ionized form (as indicators) has different colour than their neutral
molecule. They can be used for rough determination of pH. Selection of correct indicator depends on its
functional region and range of colour change. Functional region of indicator is defined as an interval
between two pH values, where we can notice colour change of indicator with a naked eye. The middle
of colour range always responds pH equal to pKA of the indicator. It applies then
functional area = pKAind ± 1
where pKAind is dissociation constant of indicator.

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Table: Examples of acid-base indicators

Indicator Colour of indicator in medium Indication region

acid base
thymol blue red yellow 1.2 – 2.8
methyl yellow red yellow 2.0 – 4.0
methyl orange orange yellow 3.3 – 4.6
methyl red red yellow 4.4 – 6.2
bromthymol blue yellow blue 6.0 – 7.6
phenolphthalein colorless violet 8.3 – 10.0
thymolphthalein colorless blue 9.3 – 10.5

Very simple is pH determination by indicator paper. Universal indicator paper with its 18-
membered comparative colour scale allows to measure pH at 0 – 12 range with possibility of estimate ±
0.5 pH.
There are several naturally existing substances in nature, their colour depends on pH of
environment. They are so-called anthocyanins, red and blue dyes, present in some flowers (e.g. rose,
geranium, mallow) and in vegetable (e.g. red cabbage). Chemically they belong to heteroglycosides
which are hydrolyzed by boiling with mineral acid to sugar component (usually monosaccharide or
disaccharide) and particular aglycone – anthocyanidin.
Chemically are anthocyanidins different hydroxy derivatives of 2-phenylbenzopyrylium chloride.

2-phenylbenzopyrylium chloride

pelargonidin cyanidin

delfinidin

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Research of anthocyanins of different flowers and fruits has led to an interesting conclusion - colours of
flowers depend on just three anthocyanidins: pelargonidin, cyanidin, and delphinidin:

These anthocyanidins and their methyl ethers give with acids and bases various colours in different
conditions. For example, blue colour of corn-flower is caused by alkali metal salt of cyanidin, while
oxonium salt of cyanidin causes red colour of rose. Pelargonidin chloride is a dye of red geranium.
The most exact measurement of pH is determination by pH-meter. It is potentiometric
determination. Potentiometry is one of the electroanalytic methods, which monitors dependence
potential of the measuring electrode against reference electrode depending on the composition of an
electrolyte. We can use potentiometry to define e.g. pH or dissociation constants.
We know direct and indirect potentiometrically determinations. Direct methods are used for
assessment the activity of some component of an electrolyte, directly from the measured equilibrium
potential. It is e.g. pH measurement. Indirect determinations qualify changes of the activity of some
solution's component using changes of measured potential.
We use potentiometers, i.e. pH-meters, for potentiometric pH measurements of the solution. pH-
meter is a voltaic cell consisting of two electrodes connected together through an electronic device.
Device is capable of measuring the potential difference between the electrodes. The electrode whose
potential does not alter with the concentration change of ions is called standard (reference) electrode.
The electrode whose potential depends on the composition of tested solution (concentration of hydrogen
ions in our case) is called measuring (indicator) electrode.

The electromotive force (EMF) of a voltaic cell equals to the potential difference of electrodes
EMF = E1 – E2,,
where E1 is the potential of measuring electrode and E2 is the potential of reference electrode.
Generally, for the electrode potential E of an electrode process (in different conditions than are
standard during equilibrium) applies Nernst-Peters equation

RT [ox]
E=E0+2,303 . log
nF [red]

where E is the electrode potential, Eº is the standard potential, n is number of electrons transferred
during electrode process, T is the absolute temperature, R is the universal gas constant, F is the
Faraday constant.
For metal electrodes with the electrode process M → Mn+ + ne-, the reduced form is metal, hence
[red] = 1. Likewise, for the electrode potential of hydrogen electrode applies

RT RT
EH2=2,303 .log[H3O+]=-2,303 . . pH=-59,2 . pH
F F

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because log [H3O+] = - pH; the value of 2.303 . RT/F is equal 59.2 mV at 25 ºC, it means when pH of
solution increases by 1 unit, the electrode potential of hydrogen electrode decreases by 59.2 mV.
Measurement of pH is in clinical biochemistry irreplaceable. pH value of body fluids gives an
information about condition of an organism. Most life processes may run only in relatively narrow range
of pH values and even slight change of this value fundamentally affects the rate of biological process,
eventually, it completely stops it. pH value of an environment most of biological processes is at range
about 6 – 9. Table shows pH values of some body and organic fluids.

Table: pH values some of body and organic fluids

Fluid pH Fluid pH
Gastric juice 1.5 – 2.0 Lemon juice 2.2 – 2.4
Urine 4.8 – 7.5 Sauerkraut 3.4 – 3.7
Saliva 5.8 – 7.1 Vinegar 2.6 – 2.7
Semen 6.9 – 7.4 Fresh milk 6.3 – 6.6
Blood 7.34 – 7.43 Tap water 5.5 – 6.0
Bile 6.2 – 8.5 Pure distilled water 7.0
Sweat 3.8 – 6.8 Beer 4.2 – 4.6

It is possible to calculate pH of every single solution by using derived formula for calculation.
Using the correct formula depends on chemical character of defined solution. Different formulas are for
pH calculation of strong acids and bases, different for pH of weak acids and bases. We will mention
formulas for pH calculation of various types of solutions.

pH calculation of strong acids and bases in solutions

Acids that almost completely dissociated in their diluted solutions are considered as strong acids.
To calculate pH we can use molar concentration, from which we can deduce concentration of hydrogen
ions. We always have to consider saturation of the acid, it defines hydrogen ions concentration. For
example, HCl dissociates HCl → H+ + Cl-, c(H+) = c(HCl), but H2SO4 dissociates H2SO4 → 2H+ + SO42-
, therefore c(H+) = 2.c(H2SO4).
To calculate the pH of a strong acid solution we use the equation
pH = - log c(H+)
Hydroxides of alkali metals and alkaline earth metals are generally considered as strong bases. It is
because of their complete dissociation into metal cation and hydroxide anion. But we cannot forget the
saturation of hydroxide – it determines concentration of hydroxide anions. For example, NaOH
dissociates NaOH → Na+ + OH-, c(OH-) = c(NaOH), but Ba(OH)2 → Ba2+ + 2OH-, c(OH-) =

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2.c(Ba(OH2)).
For calculation of pH of a strong base solution we use the equation
pH = 14 – pOH
pOH = - log c(OH-)

pH calculation of weak acids and bases in solutions

Weak acids are dissociated only partially in water solutions. The degree of dissociation
depends on value of the dissociation constant.
To find pH of weak acid solutions we need the equation
pH = ½. (pKA – log cA)
where cA is total concentration of present acid. However, it is important to realize, that pKA = -
log KA.
Analogically, to find pH of weak bases we need equation
pH = 14 – ½.(pKB – log cB)

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Laboratory Manual for Practical Exercises Acids and bases

Exercise: pH determination by acid-base indicators

Acid-base indicators are used either as solutions or as papers soaked in indicator solutions. In some
cases we use only one indicator, another time it is mixture of several indicators.
Acid-base indicators (Phan) are strips of filter paper soaked either in one or more indicator
solutions. The strip is inserted into tested solution for one second. pH determination is approximately
done by comparison of the strip colour with the colour scale on the box of indicator papers.

Materials:
Test tubes, distilled water, pipettes, indicator paper, beakers.
Solutions of CH3COOH, HCl, NH4OH and NaOH (all 0.1 mol/L), indicator solutions – methyl orange,
methyl red, phenolphtalein, methyl red.

Procedure:
1. Prepare 4 beakers with 20 mL distilled water and numbered them (tab.)

Beaker No. Water HCl CH3COOH NaOH NH4OH

1 20 2 - - -
2 20 - 2 - -
3 20 - - 2 -
4 20 - - - 2

2. Add 2 mL HCl solution into beaker No. 1, add 2 mL CH3COOH solution into beaker No. 2,
add 2 mL NaOH solution into beaker No. 3, and add 2 mL NH 4OH solution into beaker No. 4.
Mix up them.
3. The solution from each beaker divide into four test tubes (a, b, c, d).
4. Into each test tube add one drop of corresponding indicator solution by the table below.
5. Arrange all 16 test tubes (4 x 4) in such a manner that the test tubes with the same indicator
will be in a row. Compare their colours and record in the table in your note book.

Observation:
Beaker Indicator
No. Methyl orange Methyl red Neutral red Phenolphtalein
(a) (b) (c) (d)
1
2
3
4
Colour change of 3.3 – 4.6 4.4 – 6.2 6.8 – 8.0 8.3 – 10.0
the indicator (pH)

Conclusion:
What is approximate pH of acid and base solutions according to the table?

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Exercise: pH determination by natural indicators – antocyanidins

In acidic medium of mineral or organic acids antocyanidins are found as salts with red colour.
After addition of solution with pH > 7 is firstly released free antocyanidin, which is violet. Blue metals
salts of antocyanidins are formed after neutralization of all -OH groups in structure. This characteristic
can be used for pH determination.

Materials:
Test tubes, distilled water, pipettes, a leaf of red cabbage, a beaker (50 mL).
CH3COOH solution (c = 0.1 mol/L), NaHCO3 solution (c = 0.1 mol/L).

Procedure:
1. Pour 10 mL of hot water (appr. 80 ºC) into a beaker with appr. 10 g of pieces of red cabbage.
2. Extract at least 5 min with occasional stirring.
3. Transfer the extract of antocyanins into a clear beaker.
4. Prepare two test tubes and numbered them.
5. Pipette 2.5 mL of CH3COOH solution into the test tube No. 1 and 2.5 mL of NaHCO3 solution
into the test tube No. 2.
6. Add 0.5 mL of antocyanin extract into both test tubes.
7. Record in your note book the colour changes in test tubes.

Observation:

Test tube No. Colour

1
2

Conclusion:

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Laboratory Manual for Practical Exercises Acids and bases

Exercise: pH determination of body and natural fluids

pH determination of body fluids is one of important indicators of metabolic processes in organism.

For example, normal pH values of urine are in the range of 5 – 6. Urine becomes weakly alkaline only
after vegetarian diet. Because alkaline reaction of urine is exceptional it refers to some pathologic state.
Approximate determination of body and natural fluids is very simple. Commonly it is done by
universal indicator papers (e.g. Phan) stripped into a tested fluid for short time. The colour of strip is
then compared with the scale on the box.

Materials:
Test tubes, distilled water, pipettes, glass rod, litmus paper.
Samples of milk, beer, distilled water, tap water, lemon juice, urine.

Procedure:
1. Pipette 1 mL of studied sample into prepared test tubes.
2. Prepare pieces of litmus paper on laboratory desk.
3. Using glass rod dab samples of solutions on litmus paper.
4. Subtract the value of pH on the pH scale.

Observation:

Sample pH
milk
beer
distilled water
tap water
lemon juice
urine

Conclusion:

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Exercise: Measuring pH by pH-meter

pH-meter represents the electronic device designed for measuring pH. The measurement is based
on potentiometry, measuring the difference between the standard and reference electrodes. In practice,
calomel electrode is used as standard electrode and glass electrode as reference electrode.
Very common are combined glass electrodes, where glass and reference electrodes work as one
body. That electrode is composed of thin walled glass bank (0,005 mm) made of special Corning’s glass
– 72 % SiO2, 6 % CaO, 22 % Na2O, sometimes Na2O is replaced by Li2O. Inside the electrode there is
a solution of diluted HCl in which calomel or argentochlorid electrodes are dipped. pH-metres should
be calibrated before measuring by at least two standard buffer solutions with known pH.

Materials:
pH-meter, beakers, universal and Phan indicator papers, pipettes, filter paper
HCl (c = 0,1 mol/l), NaOH (c = 0,1 mol/l), CH3COOH (c = 0,1 mol/l), NH4OH (c = 0,5 mol/l),
(NH4)2SO4 (c = 0,25 mol/l), CH3COONa (c = 0,2 mol/l), Na2CO3(c = 0,l mol/l), NaHCO3 (c = 0,1 mol/l)

Procedure:
Determine pH of the solution, acids, bases and salts given by means of pH-meter, state values into the
table.

Observation:

Solutions pH determined by
universal paper pH-meter calculation
0,1 mol/l HCl
0,1 mol/l NaOH
0,1 mol/l CH3COOH (pK 4,73)
0,5 mol/l NH4OH (pK 4,74)
0,25 mol/l (NH4)2SO4
0,2 mol/l CH3COONa
0,1 mol/l Na2CO3
0,1 mol/l NaHCO3

Conclusion:
Compare the precision of pH values determined by pH meter, indicator papers and calculation.

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Buffers

In practice, there are frequently needed solutions with permanent pH value, which alternates only
a little after adding the small amount of strong acid or base. These so called buffer solutions (buffers)
are mixture of weak acids and it conjugates bases or weak bases and their conjugate acids. The typical
example is acetate buffer made of acetatic acid CH3COOH and sodium acetate CH3COONa. After
adding of the strong acid to this buffer, acetate ions made by sodium acetate dissociation bind added
hydroxonion ions H3O+ and are crossing onto nondissociated molecules of acetatic acid:
CH3COO- + H3O+ ↔ CH3COOH + H2O
When adding strong bases, acetatic acid is dissociated:
CH3COOH + OH- ↔ CH3COO- + H2O
In both cases the result is neutral H2O. This process lasts until the majority of the acetate ions or
undissociated acids molecules are consumed.
An equation relating the pH of a buffer to different concentration of conjugate acid and base is
known as the Henderson-Hasselbalch equation.
In a buffer system made up from a weak acid and its conjugate base we can get an equation:
c(HA)
c(H3O+)=KA .
c(A-)
After taking the negative logarithm of both sides:
c(A-)
pH=pKA+log
c(HA)
where KA is acid dissociation constant, pKA represents dissociated exponent (-log KA), c(A-) is the molar
concentration of conjugate base A- and c(HA) the molar concentration of weak acid HA.
Henderson-Hasselbalch equation can be expressed by the dissociation degree of weak electrolyte:
α
pH=pKA+log
1- α
where α is the level of electrolyte dissociation.
The pH value of environment affects substationally the course of chemical reactions in living
organism. A very significant value for the medicine has the acid-base balance of the inner environment
of the organism. By the regulatory function of lungs, kidneys and buffers in extracellular fluids, the
human organism is able to keep pH in a very narrow range in spite of organic acid and CO2 (around
400l) production.
From the buffer systems of blood the most important is hydrogencarbonate acid as well as
phosphate (H2PO4-/HPO42-) and protein buffers.

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Exercise: Preparation of acetate buffers

One of the best known buffers is acetate buffer system. It consists of a mixture of acetic acid and
sodium acetate solutions.
pH of the acetate buffer can be calculated by Henderson-Hasselbalch equation.

Materials:
Test tubes, pipettes, indicator paper, a glass rod, a pH-meter.
CH3COOH solution (c = 0.1 mol/L), CH3COONa solution (c = 0.1 mol/L).

Procedure:
1. Pipette CH3COOH and CH3COONa solutions into 5 test tubes by the table below.
2. Calculate pH in the test tubes by Henderson-Hasselbalch equation and record in your note book.
(KA(CH3COOH) = 1.86x10-5 and pKA = 4.73)
3. Determine pH in the test tubes by a pH-meter or the indicator paper.
4. Compare calculated and determined pH values.

Observation:

Buffer
Test tube No. CH3COOH [ml] CH3COONa pH calculated pH determined
[ml]
1 1 9
2 3 7
3 5 5
4 7 3
5 9 1

Conclusion:

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Exercise: Effect of acids and bases on pH of buffers, Buffering capacity

Buffering capacity is a quantitative measure of the buffer's ability to offset the effects of adding
a strong acid or base on the pH of a system – the resistance of a buffer solution to pH changes. By the
definition, it is a number of moles of strong acid or strong base should be added into 1 L of a buffer to
alter its pH by on pH unit. The more concentrated buffer solution, the larger buffering capacity. The
largest capacity of a buffer is when 𝑐𝐴−/ 𝑐𝐻𝐴 = 1, that means at pH = pKA.

Materials:
Test tubes, distilled water, pipettes.
Acetate buffer (c = 0.1 mol/L, pH = 4.7), 1 % solution of methyl red (a colour change in the pH range
4.4-6.2), 1 % solution of methyl orange (a colour change in the pH range 3.3-3.6), HCl solution (c = 1
mol/L), NaOH solution (c = 1 mol/L).

Procedure:
1. Pipette the solutions into four test tubes by the table below.
2. Add HCl solution by drops into test tubes No. 1 and 3 and record the volume at which is colour of
the test tubes changed.
3. Add NaOH solution by drops into test tubes No. 2 and 4 and record the volume at which is colour
of the test tubes changed.
4. Calculate the buffering capacity of the acetate buffer for acidic and alkaline legs.

Observation:

Test tube No. dist. water CH3COOH CH3COONa methyl orange methyl red
[mL] [mL ] [mL] (drops) (drops)
1 5 - - 2 -
2 5 - - - 2
3 - 2,5 2,5 2 -
4 - 2,5 2,5 - 2

Conclusion:
Make conclusions from the differences in the test tubes.

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Control questions:

1. Based on the ion product of water define the pH scale.

2. Give the relations (equations) by which we are able to calculate pH of different types of solutions.
Derive the relations for calculation of pH of: i) strong acids, ii) strong bases, iii) weak acids, iv) weak
bases and v) different types of salts.

3. Define potenciometry? What is galvanic cell (battery)?

4. Give the names and composition of blood buffer systems.

5. How is define buffering capacity?

6. How is changing the potential of the hydrogen electrode, if the pH changes of the 1 unit?

7. Define electromotive voltage article.

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8. Using a reaction explain the buffering effect of acetate buffer after adding of small amount of
concentrated nitric acid.

9. Define the difference between reference and measuring electrode.

10. Using data of table below sign the points of dependence of dissociation constant of water at given
temperature. Explain relation Kv = [H3O+] [OH-] = 1 .10-14 at 25 °C and calculate the concentration of
[H3O+].

Temperature °C 0 5 10 15 20 25 30 40 50 100
-2 -2
Kv (mol l ) .
10 14 0,114 0,186 0,293 0,452 0,681 1,008 1,471 2,916 5,476 51,3

Dependence of dissociation constant of water (Kw ) on temperature t

5,5
5
4,5
4
3,5
3
2,5
2
1,5
1
0,5
0
t °C
0 5 10 15 20 25 30 35 40 45 50

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