Chapter 2

EXPERIMENT -1

USE AND CALIBRATION OF GLASSWARE

OBJECTIVES

i. Learning to use and calibrate a pipette

ii. Learning to use and calibrate a burette

APPARATUS: Microgram balance (+ 0.0001), 10 ml pipette, weighing bottles, retort stand, and a 50 ml burette.

Introduction
Volumetric glassware, for the beginning and accomplished analytical chemist, is probably one of the most
important "tools of the trade". You will succeed or fail on your ability to effectively use glassware, which is
designed to contain or to deliver a known volume accurately and precisely. This experiment is designed to
acquaint you with volumetric glassware, both in terms of its practical use and with regard to its volume
calibration. Glass wares are calibrated to avoid introducing systematic errors into measurement.
Most take for granted the calibration of volumetric glassware. For truly exacting wet chemical analyses,
however, the importance of volume calibration cannot be overstated. As discussed in several current
quantitative analysis texts, one should compensate for temperature effects on diluent density and (in rare cases)
the thermal expansion of glass. In tandem with these effects, is the non-negligible effect of buoyancy in mass
measurements. You will learn how to compensate for thermal induced density changes for both diluent and air,
appropriate for high accuracy analytical measurements.
There are three general approaches to the calibration of volumetric glassware that are in wide use and with
which the student should be familiar.

1. The direct, absolute calibration is based on the direct mass measurement of water delivered by a burette or
pipet or contained in a volumetric flask and conversion to volume with the density, at the specified temperature.
2. The indirect, absolute calibration, or calibration by comparison, whereby a calibrated vessel is used as a
reference in the calibration of an unknown vessel, is sometimes used when many glassware items are to be
calibrated.
3. A relative calibration is based on the measured volumetric relationship between two items of glassware,
without knowledge of the absolute volume of either one. Accuracy is compromised in favor of precision.
Pipettes, burettes and volumetric flasks are intended for the most accurate measurements of solutions, but these
apparatus are not always accurate. To determine the true capacity of these instruments of precision weigh on
the analytical balance the water which they hold' or deliver . You will perform a direct, absolute calibration of a
burette or pipette in this introductory experiment. Study the following procedure and understand the
measurement techniques and calculations before coming to the laboratory for practical.
Fig. 1: Diagrams of 10ml± 0.1 transfer pipettes calibrated at 20 oC

Caution on the use of a pipette: you must first clean a pipette before using it. This can be checked by
filling the pipette with water and allowing the liquid to drain. No water drop lets should be observed on the
inner walls. THE PIPETTE IS NEVER FILLED BY MOUTH SUCTION BUT ONLY BY USING A RUBBER
SUCTION BULB OR A PIPETTE PUMP.

a b
Fig. 2 (a & b): correct use of a pipette

Fig. 3: the correct way to reading the volume of a liquid in a burette. Fig. 4: a burette
As was noted above, volumetric glassware is calibrated by measuring the mass of water that is contained in or
delivered by the device. Recalibration is usually performed by weighing the amount of water delivered by or
contained in the volumetric apparatus. This mass is then converted to the desired volume using the tabulated
density of Water:
Volume = mass / density
The volume occupied by a given mass of liquid varies with temperature, as does the volume of the device that
holds the liquid. 20oC has been chosen as the normal temperature for calibration of most volumetric glassware.
Glass is a fortunate choice for volumetric ware as it has a relatively small coefficient of thermal expansion. If
desired, the volume values (V) obtained at a temperature (t) can be corrected to 20 oC by use of:
V20 = Vt [1 + 0.00025 (20 - t)]
Where 0.00025/oC is the thermal expansion coefficient of Water.
Tolerance defines the state and quality of glass wares, the higher the tolerance values; the lower the quality of
that glass ware.
Tolerance ± (cm3) = [difference b/w nominal and measured values]
Measured values (volumes in cm3) = Vt1+Vt2+Vt3+Vt4+Vt5
5
Class A glass wares has lower tolerance values and are of more quality than Class B.

(i) Calibration of a pipette

Procedure:

1. Using a 10 ml pipette, measure and transfer 10cm 3 of distilled water into a pre-weighed clean weighing
bottle or 100 cm3 beaker.
2. Determine the weight of the 10cm3 of water and weighing bottle or beaker.
3. Repeat this experiment five times using the same volume of water.
4. Record the temperature of the distilled water also for each time with a mercury thermometer. Using the
density of water at the temperature of this experiment (table 4), hence, calculate the volume of each
mass of the water obtained.
5. Convert this volume to standard volume at normal calibration temperature.
6. Calculate the relative standard error of the pipette at 20oC.
7. Find the mean volume and Compare this volume with the delivering volume of burette.
8. Which of the two is more accurate?
9. Determine the tolerance of the pipette.

Table 1: Standard calibration table for pipette at 20oC

Volumes (cm3) Class A Class B

5 0.02 0.03
10 0.03 0.02
25 0.04 0.03
50 0.06 0.04
100 0.06 0.10
200 0.20 0.10
250 0.20 0.10
500 0.30 0.15
1000 0.40 0.20
2000 0.60 0.40

Table 2: Tolerance value for various volumes of Pipettes

Capacity (ml) Tolerances (ml)

0.5 0.006
1 0.006
2 0.006
5 0.01
10 0.02
20 0.03
25 0.03
50 0.05
100 0.08

Table 3: Tolerance value for various volumes of burettes

Capacity (ml) Tolerances (ml)

5 0.01
10 0.02
25 0.03
50 0.05
100 0.20

(ASTM E694)

Result and calculations:

Temperature of distilled water = t oC

Density at t oC temperature =

S/N Mass of empty Mass of empty Mass of water (g) Volume of water cm3
weighing bottle (g) weighing bottle +
water (g)
1
2
3
4
5

√
2 2 2
( x1 − x̄ ) + ( x 2− x̄ ) + ( x 3 − x̄ ) +. . .. .. . ..
Hint: Standard Deviation (S.D) = N −1

Where x 1 …. = individual observations, x̄ = mean, N = total no. of observations

S. D
Relative Standard Error = √ N

Where S.D = Standard Deviation, N = No. of observations

(ii) Calibration of a burette

Procedure: It is, of course, practically impossible to check the correctness of all the 500 or 250 divisions on a
burette. Usually, the corrections of only the marks which correspond to the volumes of 5, 10, 15ml and so
forth, is checked. The burette is checked as follows.

1. Fill a clean burette with distilled water.

2. Fix the water level at exactly zero division.
3. Pour out water three times up to the division into a pre -weighed weighing bottle or 50ml beaker and
weigh it.
4. Calculate the correction for every checked division.
5. Draw up a table on the basis of the data obtained and plot the correction curve.
6. Use the correction curve to correct for the unchecked divisions by extrapolation.
Table 4: Density of water
Tutorial Questions.

1. When the glass of a burette expands due to an increase in temperature does the diameter of the bore increase
or decrease?
2. Most volumetric glassware is calibrated at what temperature?
3. What do the letters T.D. and T.C., which are found on various types of volumetric glassware, signify?
4. A dilute aqueous solution with a molarity of 0.0314 M was prepared in the winter when the laboratory
temperature was 17oC. What will be the molarity of that solution on a warm spring day when the temperature is
25oC?
Note: Concentration is directly proportional to density (C α D)
5. A pipette was calibrated to deliver 25 cm3 of water into an empty weighing bottle of 10.283g at room
temperature of 23oC. What will be the volume at standard calibration temperature?
6. The following values were obtained for the calibration of a 10-ml pipet: 10.10, 9.98, and 10.00 ml. Calculate
the mean value and the average deviation from the mean.

Reference materials

1. Quantitative Chemical Analysis, 7th Ed., 2007, by D.C. Harris (Chapter 2)

2. Analytical Chemistry 6th Ed. by G.D. Christian (2004) Wiley, (Chapter 2)

Chapter 3

INTRODUCTION TO TITRIMETRIC TECHNIQUES

Quantitative analysis determines the amounts of the elements or compounds present in a sample. This is usually classified
on the basis of different techniques of reaction such as gravimetric, volumetric and instrumental analysis. If the analysis is
carried out solely using solutions of chemical substances, this is called a classical analysis or wet chemistry reaction e.g.
gravimetric and volumetric analysis but if the analysis is performed using a device, then it is called instrumental analysis.
Some important techniques utilizing this method are spectroscopic analysis, electrochemical analysis, chromatographic
analysis, colorimetric analysis, nuclear magnetic resonance etc. one of the important methods in quantitative analysis is
volumetric analysis, a commonly used laboratory techniques.

Volumetric analysis is a quantitative analysis or techniques based on finding the volume of one solution which will
exactly reacts with a definite volume of another solution of known concentration. The reaction employed in volumetric
analysis can be classified into two:

I. Those in which no change in oxidation state occurs; these are dependent upon the combinations of ions which
includes neutralization reactions, complex formation reactions and precipitation reactions.
II. Those that involves a change of oxidation state; oxidation-reduction reactions.

In volumetric analysis, the concentration of analyte can be found by reacting it with a standard solution of known
concentration. Volumetric analysis is based on the stoichiometry between the reactive species.

e.g HCl + NaOH → NaCl + H2O

Acid Base Salt Water

The concentration of the acid can be found by reacting it with the base of exactly known concentration (standard
solution). In this method, a known volume of one solution is taking in conical flask and solution of other substance is
added very slowly to it till the reaction is complete. From the volume of the unknown solution which is required to
completely react with definite volume of the other solution of known concentration, the concentration of the unknown
solution can be calculated using the law of equivalence which states that the substances react with each other in a definite
ratio by mass. If an acid is titrated with a standard base, then at the end point according to normality equation:

No. of gram equivalents of acid = No. of gram equivalents of the base

NAVA =NBVB where

NA is the Normality of the acid,

VA is the volume of the acids,

NB is the Normality of the Base, and

VB is the volume of the Base.

Background theory:

Acid base titration involves a neutralization reaction. In this titration, acids are determined by titration with an appropriate
standard base solutions and bases by titration with an appropriate standard acid solution. From point of view of analytical
chemistry, all chemical substance may be divided into two categories.

Category 1: substances whose purity and properties have been exactly known and verified. They are referred to as
primary standards. Primary standards are highly purified chemical compounds used as reference materials in preparing a
standard solution.
Category 2: Substances whose purity cannot be exactly known mainly because they are highly unstable, hygroscopic or
because they are volatile. It is easy to prepare a standard solution from group I compounds.

The substances is simply accurately weighed and dissolved to a known volume in a volumetric flask. Commonly, the
standard titrant solutions are prepared from group two substances as solution of approximate concentration then they are
titrated against a solution of a primary standard to determine their exact concentration. This procedure is referred to as
standardization. For instance, if you want to prepare a titrant solution of NaOH, you cannot weigh directly NaOH pellets
because NaOH is hygroscopic i.e. it absorbs unspecified amount of water from air and may be covered with a thin layer
of sodium bicarbonate due to reaction with carbon dioxide from air. The only way to solve this problem is to prepare an
approximate concentration by roughly weighing NaOH pellets and dissolving in approximate volume of distilled water
and then titrate the resulting solution with a primary standard of known concentration. This procedure is referred to as
standardization. Standardization usually involves:

-Preparation of a standard solution.

-Preparation of solution of approximate concentration to be standardized.

-Titration: the process during which exact concentration of the solution to be standardized is determined.

-Calculation of the result from data obtained in titration.

In the titration process one solution usually the standard (titrant) is slowly added from the burette to another solution
placed usually in a conical flask (the titrand) that contain a known volume or known mass of solute until the chemical
reactions between the solute is complete. The point at which stoichiometrically equivalent quantities of substances have
been brought together is known as equivalent point of titration. To determine when equivalent point of titration is
reached, an indicator is used. In acid base titration, the indicator used are usually organic dyes. For example
phenolphthalein is the dye that is colourless in acidic solution and pink in basic solution.

In fact, it is rarely possible to find an indicator that will indicate exact equivalent point. Hence the change in colour we
observed when we stop the titration indicates the end point of titration. Although the end point of titration does not
coincide exactly with the equivalence point the error is negligible.

Standard solution

A standard solution is a solution containing a precisely known concentration of a substance. A known weight of solute is
dissolved to make a specific volume. A standard solution can be prepared from primary or secondary standard substances.
A primary standard is any substance whose standard solution can be prepared directly by dissolving an accurately
weighed amount of it in water and making up the solution to a known volume by dilution with water. It is highly pure,
high molecular weight chemical compound, stable with no water of hydration e.g Na 2CO3, AgNO3, KCl,
FeSO4(NH4)2SO4.6H2O etc. In secondary standard, an approximate weight of the substance is dissolved in a known
volume of the solvent. The exact concentration of the solution is found by titrating it against a primary standard e.g
NaOH, KMnO4, KOH, HCl, H2SO4 etc.

Solution preparation
Preparing solution in the laboratory is an exercise considered to be tedious by student because of the various
techniques, skills and manipulations it involves. What student finds most difficult is on how to know the exact
mass or volume of substance to be dissolved in aqueous solutions, the concentrations to be prepared, dilutions
and calculations regarding solution preparation. But rather than shying away from it, you could however
become a wonderful scientist by learning the techniques of solution preparation. It is one of the skill you need
to excel as an analytical chemist.

Usually, most laboratory chemicals are commercially packaged as liquid or as solid substances. To prepare the
solution of this chemicals, proper knowledge and understanding of the information provided on the label of
chemicals could be very useful. Chemical labels carries information on the molecular weight of the compound,
the specific gravity, the percentage purity or percentage composition (minimum assay), maximum limits of
impurity and safety information which are very crucial to solution preparation.

To prepare a standard solution of a solid chemical, you first need to calculate the molecular weight of the
compound. Check up the information on the label.

Solid chemicals compounds with unknown molarity: For example to prepare 1M solution of sodium sulphate
Na2SO4 with molecular weight of 142.04g/mol, you will need to weigh this exact mass (142.04g) and dissolve
in 1litre of distilled water in a volumetric flask as shown in the diagram below.
From Basic Laboratory Methods for Biotechnology: Textbook and Laboratory Reference,
Seidman and Moore, 2000

If the concentration you need is 0.1M. The previous concentration has been reduced by a factor
of 10, 1/10 = 0.1. Then divide the mass of Na 2SO4 (142.04g) by 10. This will give 14.204g.
Weigh 14.204g and dissolve in 1 litre of distilled water as 0.1M solution of Na 2SO4. You can
always prepare a solution of any solid chemical compounds going by this rule.

Liquid chemical compound with unknown molarity (stock solution): The concentration of
commercial aqueous reagents are often express by weight percent. Thus trioxonitrate (V) acid is
sold as 70% solution which means that the reagent contains 70grams of trioxonitrate (V) acid per
100 grams of solution. To prepare this kind of solutions, an understanding of density, mass
percentage, conversion of mass to moles, molarity and dilution law are required.

Example: Describe the preparation of 200cm 3 of 6M HCl from the concentrated reagent. The
label on the bottle states that the reagent is 37% HCl and has a specific gravity 1.18.
Solution: The specific gravity of 1.18gcm -3 can be interpreted as; 1 cm 3 of the acid contains
1.18g HCl.

If it is 37% pure HCl, therefore, 1cm3of the acid will contain 37/100 x 1.18g

= 0.44g of HCl.

Next, convert the mass to mole.

Molar mass of HCl = 1+ 35.5 = 36.5gmol-1

Note, amount in mole = Mass

Molar mass

1cm3 of the acid contains 0.44g/36.5gmol-1 = 0.0120 mol of HCl.

1 dm-3 or 1000cm3 of the acid will contain 0.0120 x 1000 =12 M HCl

Using dilution law derivable from the law of conservation of matter,

C1V1 = C2V2

This equation applies to all dilution problems.

C1 (initial conc.) x V1 (initial volume) = C2 (final conc.) x V2 (final volume)

C1 = 12 M
V1 = unknown
C2 = 6 M
V2 = 200 cm3

V1 = (C2 x V2) / C1

= (6 X 200) / 12
= 1200 / 12 = 100 cm3.
This means that 100 ml of the 12 M HCl stock solution diluted to a final volume of 200cm3 with
distilled water in a standard flask will give a concentration of 6 M HCl.

Strength of a solution

This is defined as the amount of substance dissolved in one litre of a solution when expressed in
grams is termed as strength of a solution. Strength of a solution can also be expressed in terms of
Normality, Molality, Molarity, Formality etc.

Methods of expressing concentrations

Concentrations can be defined as the amount of solutes dissolved in a given quantity of solvent
or solution. The concentration of solution can be expressed either qualitatively or quantitatively.
The term dilute and concentrated are used to quantitatively describe a solution. A solution with a
relatively small amount of solutes is said to be dilute, while one with a large amount of solute is
said to be concentrated.

Molarity: This is the number of moles of a solute dissolved in a litre of solution. It is a

concentration term and its temperature dependent.

Molarity (M) = Moles of solute

Litres of solution

Molality:
The molality of a solution is defined as the number of moles of solute in one kilogram (kg) of
solution. It is a composition term (i.e mass of solute per unit mass of solvent), it is useful for
precise physical measurements. The reason is that molality is not temperature dependent,
whereas molarity is temperature dependent.
Molality = No. of moles or solutes
Kilograms of solvent

It can also be defined as the number of gram molecular masses of solute per 1000 grams of
solvent. A solution which contained 58g of NaCl in 1000g of water would be a 1Molal solution
(1m).

Formality: This is also a composition terms. It is expressed as the number of gram formula
masses of solute per kilograms of solvent.
Formality = Formula masses of solute
Kilogram of solvent
A solution which contain 58g of NaCl in 1 kg of water would be a 1 Formal solution (1F).

Normality: Normality is the number of gram equivalent of solute per litre. A 1N solution is a
solution containing one equivalent weight per litre thus, the normality of a monovalent
compound is the same as its molarity. Equivalent of a substance is the mass of that substance that
will combine with 1g atom of hydrogen. For example for H 2SO4, its equivalent mass is half mole
(i.e 2H+ are replaceable).

Normality = gram equivalent of solute

Litre

Osmolarity: Osmolarity is the total number of moles of particles dissolved in one litre of
solution. For non-electrolyte, the osmolarity is equal to the molarity but for strong electrolyte
like CaCl2, the osmolarity is 3 times the molarity, since each formula weight CaCl 2 provides 3
2+
moles of ion in solution (Ca 2Cl). Osmolality is the number of moles of solute per kilograms
of solvent.

Part per million: 1 part per million indicates that one gram of substance of interest is present
per million grams of total solution or mixture. For an aqueous solution whose density is close to
1g/ml, 1 ppm = 1µg/ml or 1mg/l
Parts per million (ppm) = Mass of solute × 106
Mass of solution

Parts per billion (ppb) = Mass of solute x 109

Mass of solution

Mole fraction: mole fraction (χ) is the moles of solute per total moles of components present in
the solution. If a solution contains component A, B, C, then the mole fraction of component A

χA = nA
nT
nT = nA + nB + nC
χA+ χB + χC = 1 ( unity)

Weight percentage (%): Weight percentage is defined as the ratio of the weight of solute to the
total weight of solute plus solvent multiplied by 100 or grams of solute in 100 grams of solution.
Wt/wt % = wt of solute x 100
wt of solute + wt of solvent

Wt / vol % = Weight of solute in gram x 100

Volume of solutions in cm3
This can be express as the grams of solute in 100ml of solution that is why a 10% sodium
chloride contains 10g of the salt in 100ml of water.

Volume percent: Volume percent is the volume (ml) of solute per 100 ml of solution
Vol / vol % = Volume of solute in cm3 x 100
3
Volume of solutions in cm

10 cm3 of ethanol plus 90 cm3 of water is a 10 % by volume solution.

Volume ratios: It expresses the amount in volumes of each parts or components of the solution
in ratios. For example a solution of 3:2:1 ethylene: chloroform: isoamyl alcohol contains 3 litres
ethylene, 2 litres chloroform and1 litre isoamyl alcohol respectively. Concentration can also be
expressed using mass or molarity ratio.

Calculating molarity from gram concentration:

Molarity can be calculated from gram concentration by using this formula
Concentration in mole per dm3 (Molarity) = Concentration in gram per dm3
Molar mass
 THEORY OF INDICATORS

An indicator is a chemical compound that undergoes a definite colour change at a specific

electrode potential. An acid/base indicator is an inorganic compound, which is added to an acid
to indicate when the end point is reached. It is either a weak acid or a weak base and can
therefore take part in equilibrium of the type

HIn(aq) → H+(aq) + In–(aq)

The unionized form (HIn(aq) has a different colour from the ionized form (In -). The colour of an
otherwise uncolored solution in which it is put will; therefore depend on the ratio of the two
forms, which in turn depends on the pH of the solution. Since pH changes during titration, the
colour of the solution containing the indicator will also change in the course of the titration.

With the appropriate choice of indicator, the colour change can be made to coincide with the end
point of acid/base titration.

When the two differently colored forms of an indicator are present in equal amounts, it gives an
intermediate colour that is often called the “neutral point” of the indicator. The “neutral point”
of an indicator does not always correspond to a pH of 7.

The particular pH at which an indicator displays its neutral colour is called the pK of that
indicator, where K is the acid dissociation constant of the indicator. If pK is less than 7,
indicator will show its basic colour and if pK is greater than 7, it will show an acidic colour.
Some indicators used in titration are KMnO4, n-Phenyl Anthranilic acid, Starch,
Phenolphthalein, Methyl orange, Congo red, Alizarin R., Bromothymol blue, Phenol Red,
Bromocresol purple etc.
End point and equivalent point: The end point of a titration is the point at which the reaction
between the titrant and the analyte becomes complete usually determined using an indicator.

Tutorial questions:

1. Determine the colour of the acidic form and of the basic form for each of the following
indicators:

 Methyl orange
 Congo red
 Phenolphthalein
 Bromothymol blue
 Bromocresol purple
 Alizarin R.
 Phenol red
 Cresol red.
2. How many grams of potassium oxalate should be dissolved in 500ml to make a 0.1N solution
for the titration of MnO4- as shown in the reaction below?
5H2C2O4 + MnO4- + 6H+ ↔ 2Mn2+ +10CO2 + 8H2O
3. A sample of salt water with a density of 1.02g/ml contains 17.8ppm nitrate NO 3-. Calculate the
molarity of nitrate in water.
4. A solution contains 2.65g of anhydrous NaCO3 in 250 ml of solution.
Find the concentrations of the solution in:
i. Grams per dm3
ii. Weight / weight percent
iii. Weight/ volume percent
iv. Mole per dm3
(The formula weight of Na2CO3 = 106, l ml of water weighs 1g.)
5. You have available 12.0 M HCl (Conc. HCl) and wish to prepare 0.500 litre of 0.750 M HCl
for use in an experiment. How do you prepare such a solution?
6. Describe the preparation of 500cm 3 of 6 M H3PO4 from the commercial reagent that is 85%
H3PO4 and has a specific gravity of 1.69.
7. How would you prepare 500 mL of a 5 % (w/v) solution of NaCl?
8. List 5 common indicators used in acid –base titration.
9. State the difference between end point and equivalent point
10. What is the principle of volumetric analysis?
11. Differentiate the following i) acid-base titration and precipitation titration ii) primary
standard and secondary standards iii) titrant and titrand.
12. On which factor is the use of acid-base indicator in titration depends
13. Why is KMnO4 not a primary standard and how can it be standardized?

Reference

Jack T. Ballinger, Gershon J. Shugar (1996): Chemical technicians’ ready reference hand book
4th edition, McGraw‐Hill Inc., New York, pg 391, 419-424. ISBN 0–07–0571 86–4.