BP102T.

PHARMACEUTICAL ANALYSIS (Theory)

UNIT-1
a) Pharmaceutical Analysis - Definition and scope
i. Different techniques of analysis
ii. Methods of expressing concentration.
iii. Primary and Secondary standards.
b) Errors: Sources of errors, types of errors, methods of minimizing errors,
accuracy, precision, and significant figures.

PHARMACEUTICAL ANALYSIS

Pharmaceutical Analysis is a branch of chemistry that deals with the separation,

identification, determination, quantification, and purification of a substance in a
sample.

OR

Pharmaceutical Analysis is a branch of chemistry that involves the application of

analytical procedures to ensure the purity, quality, and safety of pharmaceutical
products.

It involves qualitative and quantitative analysis that consists of a series of

procedures for identification, determination, quantitation, and purification.

The major applications of pharmaceutical analysis include-

✓ Identification of chemical categories of compounds,

✓ Determination of compounds in mixture
✓ Separation of the components from a mixture.
✓ Isolation, purification, and structural identification of compounds.

Pharmaceutical Analysis and quality control are vital in the pharmaceutical

industry as pharmaceuticals have an important role in human health.

Quality of drugs is vital and those that deviate in quality aspects are referred to as
sub-standard. The quality of the drug (product) is an output of a series of analysis
starting from the raw materials, in-process during the conversion stage, and

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extending till the finished product. The quality of the product should be ensured in
every stage of manufacturing and the quality of the drug should be in accordance
with the standard protocol prescribed by pharmacopoeia.

TYPES OF PHARMACEUTICAL ANALYSIS:

1. QUALITATIVE ANALYSIS
2. QUANTITATIVE ANALYSIS

1. QUALITATIVE ANALYSIS: It may be defined as “which analyte is

present in the given sample”. This has the goal of identifying the various
components present in the given sample based on physical and chemical
properties such as functional groups, elemental composition melting point,
etc.

2. QUANTITATIVE ANALYSIS: It may be defined as “How much amount

of analyte is present in the given sample”. This has the goal to determine the
quantity of each component present in the given sample.

SCOPE OF PHARMACEUTICAL ANALYSIS:

1. In Pharmaceutical Industry

2. In the food Industry

3. In cosmetic industry

4. In disease diagnosis

5. In Soil Study

6. Environmental pollutant determination

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1. In the Pharmaceutical Industry: There are different sectors in the
pharmaceutical industry as research and development (R&D) and Quality
control (QC) in which pharmaceutical analysis is utilized regularly.

2. In the food Industry: As we all know, packed food consumed by

consumers should have all parameters like quantity, purity, and safety which
enhance acceptability by consumers. For this, it is required to analyze all
these parameters for packed food. Different kinds of preservatives, coloring,
flavoring, and sweetening agents are used in packed food which can be
natural or synthetic chemical ingredients so these should be analyzed
qualitatively and quantitatively, for these various kinds of analytical
techniques are used.

3. In the cosmetic industry: The preparation of cosmetics, such as lipsticks,

creams, nail paints, lotions, shampoo, and conditioners, etc., play with two
things color or odor, and these coloring agents’ fragrances are also built by
different chemical ingredients so the quality and quantity of these
ingredients should be known which can be analysed by different techniques
of analysis.

4. In disease diagnosis: Different diseases can be diagnosed by

pharmaceutical analysis techniques like HIV is observed by the ELISA
method.

DIFFERENT TECHNIQUES OF ANALYSIS

1. Volumetric or Titrimetric Analysis

2. Electrochemical Analysis
3. Spectroscopic Analysis
4. Chromatographic Analysis

1. Volumetric Analysis: Volumetric methods include the determination of the

volume or weight of the standard reagent required to react completely with
the analyte.

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 As the name is clear in this titration volume is used. So, it is a quantitative
method.

Volumetric Analysis is classified into:

a) Acid-base titration
i. Aqueous titration
ii. Non-aqueous titration
b) Redox titration
c) Complexometric titration
d) Precipitation titration

2. Electrochemical Analysis: Electrochemical analysis is an analytical

technique that uses a measurement of potential, charge, or current to
determine an analyte’s concentration.
a. Potentiometric
b. Conductometric
c. Amperometric
d. Polarography

3. Spectroscopic Analysis: Spectroscopic methods are widely used for

qualitative and quantitative analysis. They give results in the form of an
interpretable spectrum and therefore are called spectroscopic methods.
Spectroscopy can involve any interaction between light and matter,
including absorption, emission, scattering, etc.

Four techniques are commonly used to help with structural analysis:

a) Ultraviolet-visible spectroscopy
b) Infrared Spectroscopy (IR)
c) Nuclear Magnetic Resonance Spectroscopy (NMR)
d) Mass Spectroscopy

4. Chromatographic Analysis: Chromatographic methods are mainly used for

the determination of complex mixtures or mixtures of components by
separating them into individuals.

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 a) Gas chromatography
b) High-performance liquid chromatography
c) Thin-layer chromatography
d) Column Chromatography

METHODS OF EXPRESSING CONCENTRATION

In all the techniques of quantitative analysis, the use of solutions requires some
basis for the expression of solution concentration. All the systems of concentration
expression have a fundamentally similar basis with respect to weight relationships
of solute and solvent, but the actual method of expression of concentration should
take on some convenient and specific form.

The concentration of an analyte in a solution can be expressed by more than one

method depending upon the type of analysis carried out.

Major methods for expressing concentration are: -

a) Percent Concentration
b) Normality
c) Molarity
d) Molality
e) Formality

a) Percent Concentration: Concentration is many-a-times expressed in terms of

percent (parts per hundred). The percent composition of a solution can be
expressed as:

i) Percent w/w = Mass of solute x 100

Mass of solution

ii) Percent v/v = Volume of solute x 100

Volume of solution

iii) Percent w/v = Mass of solute x 100

Volume of solution

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Percent w/w is frequently employed to express the concentration of commercial
aqueous reagents, percent v/v is used to specify the concentration of a solution
prepared by diluting a pure liquid with another liquid and percent w/v is employed
to indicate the composition of dilute aqueous solutions of solid reagents.

b) Normality: The normality of a solution can be defined as the number of grams

equivalent of solute dissolved in 1000ml of solution. It is denoted by ‘N’.
𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑔𝑟𝑎𝑚 𝑒𝑞𝑢𝑖𝑣𝑎𝑙𝑒𝑛𝑡𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
𝑁𝑜𝑟𝑚𝑎𝑙𝑖𝑡𝑦(𝑁) =
Volume of solution in litres

Equivalent Weight of an acid: It is the ratio of molecular weight to its basicity.

𝑀𝑜𝑙𝑒𝑐𝑢𝑙𝑎𝑟 𝑤𝑒𝑖𝑔ℎ𝑡
𝐸𝑞𝑢𝑖𝑣𝑎𝑙𝑒𝑛𝑡 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑎𝑐𝑖𝑑 =
basicity
Basicity is the number of replaceable hydrogen atoms present in an acid.

a. Example: - Equivalent weight of hydrochloric acid is: -

36.5 36.5
𝐸𝑞𝑢𝑖𝑣𝑎𝑙𝑒𝑛𝑡 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝐻𝑐𝑙 = = = 36.5
basicity 1

b. Example: - Equivalent weight of Sulphuric acid: -

98 98
𝐸𝑞𝑢𝑖𝑣𝑎𝑙𝑒𝑛𝑡 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝐻2𝑆𝑂4 = = = 49
basicity 2

Equivalent weight of base: It is the ratio of molecular weight to its acidity.

𝑀𝑜𝑙𝑒𝑐𝑢𝑙𝑎𝑟 𝑤𝑒𝑖𝑔ℎ𝑡
𝐸𝑞𝑢𝑖𝑣𝑎𝑙𝑒𝑛𝑡 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑏𝑎𝑠𝑒 =
acidity

Acidity is the number of replaceable hydroxyl groups of the base.

a. Example: - Equivalent weight of sodium hydroxide is: -

40 40
𝐸𝑞𝑢𝑖𝑣𝑎𝑙𝑒𝑛𝑡 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝐻𝑐𝑙 = = = 40
acidity 1

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b. Equivalent weight of calcium hydroxide: -
74 74
𝐸𝑞𝑢𝑖𝑣𝑎𝑙𝑒𝑛𝑡 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝐶𝑎(𝑂𝐻)2 = = = 37
acidity 2

c) Molarity: Molarity can be defined as the number of moles of solute dissolved in

1000ml of solution. It is denoted by ‘M’. We need two pieces of information to
calculate the molarity of a solute in a solution: -

The moles of solute present in the solution.

The Volume of solution (in liters) containing the solute.

To calculate molarity, we use the following equation: -

𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
𝑀𝑜𝑙𝑎𝑟𝑖𝑡𝑦(𝑀) =
Volume of solution in litres

c) Molality: It is defined as the number of moles of solute dissolved in 1 kilogram

of solvent. We need two pieces of information to calculate the molality of a solute
in a solution: -

The moles of solute present in the solution.

The mass of solvent (in kilogram) in the solution

To calculate molality the following equation is used: -

𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
𝑚𝑜𝑙𝑎𝑙𝑖𝑡𝑦(𝑚) =
mass of solvent in kilogram

d) Formality: The formality of a solution may be defined as the number of gram

formula masses of the ionic solute dissolved per liter of the solution. It is denoted
by F. Commonly; the formality term is used to express the concentration of the
ionic solids which do not exist as molecules but exist as networks of ions. A
solution containing one gram formula mass of solute per liter of the solution has

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formality equality to one and is called formal solution. The formality of a solution
changes with change in temperature.
𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑔𝑟𝑎𝑚 𝑓𝑜𝑟𝑚𝑢𝑙𝑎 𝑚𝑎𝑠𝑠𝑒𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
𝐹𝑜𝑟𝑚𝑎𝑙𝑖𝑡𝑦(𝐹) =
Volume of solution in litres

Primary and Secondary standards

Standardization:

Standardization means the determination of the concentration of a given sample

solution and can be determined by reacting the solution quantitatively with a
standard solution.

Very pure reagents of high stability are used in the preparation of standard
solutions. In such cases the accurate weight of the reagent is taken, dissolved, and
diluted to the exact known volume, and concentration is calculated on a theoretical
basis.

a) Primary Standard:

The substances of high purity used in the preparation of standard solution are
known as primary standard substances. A solution with an accurately known
concentration is called a primary standard solution.

The primary standard should satisfy the following requirements:

a) It must be easy to obtain, purify, dry, and preserve in a pure state.

b) It should be 100.00% pure although 0.01 to 0.02% impurity is tolerable if
accurately known.
c) It should be stable in atmospheric conditions. It should not decompose or
hygroscopic.
d) It should be readily soluble under the conditions in which it is to be
employed.
e) The substance should be unaltered in the air during weighing.
f) It should have a high relative molecular weight so that the weighing errors
may be negligible.

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b) Secondary Standard:

A secondary standard is a substance used for standardization and whose

concentration has been determined by comparison with the primary standard.

Properties

a) It has less purity.

b) Less stable
c) More reactive
d) But its solution remains stable for a long time.
e) Titrated against a primary standard.

ERRORS

In pharmaceutical analysis, the aim of an analyst who is performing an analytical

operation/experiment in the analytical laboratory must be that he or she must
obtain a true result by performing an analytical procedure correctly as required.
But in reality, an analyst who knows the various aspects of analytical operations
knows very well that the true or theoretical value of the result is very rare, and
he/she should only try to get his/her results close to the true value. How close is the
result to the true value depends upon the knowledge of the analyst towards the
chemistry or principle of operation, possible interferences in operation i.e., from
other ions, elements, and compounds as well as hands-on experience with the
statistical distribution of values.

“The term error refers to the difference between the measured value and true value
in the results of any analytical operation”.

However, it is not possible to eliminate error from any measurement, even if the
person operating is an expert, performing the operation in best condition and
performing on the best quality instrument. One should only make attempts to
minimize errors and get results that are close to the true value at the best possible.

Example: If a tablet contains 500mg of paracetamol and after analysis the analyst
observed 510mg of paracetamol tablet.

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Then,

Absolute Error = observed value – True value

510-500 = 10mg

% Relative Error = Observed value – True value x 100

True Value

510-500 x 100 = 2%

500

Types of Errors:

Errors that occur in analysis can be majorly classified into two types:

1. Determinate Errors
2. Indeterminate Errors

1. Determinate errors:

It is also known as systemic error. These are the errors whose source or cause as
well as magnitude is known and can be avoided or minimized by following proper
procedure carefully and taking necessary precautions.

Systemic errors can be classified as follows:

a) Personal Error
b) Instrumental Error
c) Reagent Error
d) Method Error
e) Proportional Error
f) Constant Error

a) Personal Error: For these errors, the person carrying operation or

experiment is responsible. These errors are common when the analyst not
personally doing the operation but is connected to the procedure as an
observer.

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Some of the examples of personal errors are: -

• The person carrying titration cannot be able to detect the endpoint by color
change clearly and always add more amount of titrant than required which
results in a positive error.

• The person carrying out titrations is unable to read the burette reading
properly and always notes incorrect values that also cause an error in results.

b) Instrumental Error: Instrumental Errors are seen when the instrument used
in the operation doesn’t give the accurate measurement that it needs to be
given. These are due to the use of uncalibrated glassware like pipettes,
burettes, etc.

c) Reagent Error: Reagent errors are due to the reagents used in the analysis.
These errors are seen when contaminated reagents are used in the analysis,
or the reagents attack on the apparatus or glassware and introduce impurities
in the analyte sample under analysis.

d) Method Error: These errors are not easy to detect which usually occur
when the analytical method is not performed properly or is unsuitable for
particular analysis.

e) Proportional Error: The magnitude of proportional error depends upon the

amount of sample. Proportional errors are due to the materials that interfere
with the analytical procedures. For example, the use of impure NaOH in the
titration of HCl gives an error in results and the concentration of NaOH is
proportional to the error in the result.

f) Constant Error: The constant error is independent of the magnitude of the

measured amount but becomes less significant as the magnitude increases.
For example, in titrations addition of 0.1 ml more titrant in 10ml titrand to
see color change clearly at the endpoint gives an error of 0.1ml. Similarly for
20 ml titrand 0.1 ml more titrant as added it also gives the same 0.1 ml error
even on double the size of the sample. Therefore, as the amount of titrand
increases the significance of constant error decreases.

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2) Indeterminate Error: It is also known as random error. The cause of random
error is usually not known therefore cannot eliminated by careful working or
taking precaution during analysis. For example, a person performing titration by
same method in same conditions for same sample, the results are not identical. The
error observed in that condition is called random error.

SOURCES OF ERROR:

Errors in the result in an analysis can be resulted from various sources. Some major
sources of errors in pharmaceutical analysis are described here –

1. Human Source

2. Instrumental, apparatus and glassware

3. Experimental condition

4. Constituents used in analysis.

5. Procedure

1. Human Source: The qualification and experience of an analyst performing the

analysis has a major impact on errors in results. If an experiment is performed by
an inexperienced person, the chance of error is higher as compared to the same
experiment performed by the experienced person.

2. Instrumental, apparatus, and glassware: If the instrument, glassware as well

as apparatus used in analysis is of low quality and uncalibrated the chances of error
are increased at significant extent.

3. Experimental Condition: If the analysis is carried out in the conditions which

are unfavourable for particular experiment or analysis the desirable result will not
obtained.

4. Constituents used in analysis: If various constituents like standard, solvent,

reagents etc. used in analysis are not of desired quality and purity the results will
obtained with errors.
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5. Procedure: If the analytical procedure used in analysis is not validated and if
validated but not followed carefully the errors in the results will be obtained.

METHODS OF MINIMIZING ERRORS

As we know the error is an in differentiate part of any analytical operation.

Analysts try his best to minimize the error as much as possible. Errors can be
minimized by applying following methods during analysis: -

1. Proper calibration of instruments, apparatus, and applying corrections

2. By running control determination

3. By running blank determination

4. By comparing the results of independent methods

5. By using the standard addition

1. Proper calibration of instruments, and apparatus and applying corrections:

By calibration of instruments like U.V. potentiometer, pH meter, etc. and glassware
like burette, measuring cylinder, pipette, etc. before performing analysis one can
eliminate errors to a very much extent. Errors can be also minimized by applying
necessary corrections if the source of error is known.

2. By running control determination: By running control determination

parallelly to the sample by taking standard under the same experimental conditions
the error can be reduced to a very possible extent. However, a standard should
contain the same weight of the constituent present in the unknown sample.

3. By running blank determination: Blank determination is the determination

under the same experimental conditions that are used for sample analysis but in
this case, the sample is excluded. Blank determination aims to know the effect of
impurities introduced by eliminating the error to a very much extent.

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4. By comparing the results of independent methods: In that case, the analysis
of same sample by totally different method is performed and the results obtained
from both methods are compared. If they are identical then they are considered as
reliable. For example, Determination of concentration of HCl solution is performed
by titration with standard NaOH and also by precipitation titration method with
AgCl2 and result obtained from both methods are compared.

5. By using Standard addition: In this method, small amounts of standard

constituents present in sample is added and sample is analyzed for the total amount
of constituents present. The difference in the results from sample with and without
adding a standard is calculated. Satisfactory recovery gives the confidence of the
accuracy of the result.

ACCURACY:

The term “accuracy” refers to the concordance between the measured value and
true value or in other words it refers to how close the measured value is to the true
value. It also refers as “correctness of measurement”. In relation to error accuracy
is inversely proportional to it i.e. more the accuracy less will be the error or vice-
versa.

PRECISION:

Precision refers to the degree of closeness between the several measurements of

same quantity of sample by same method.

Precision and accuracy are correlated to each other. Precision always represents
accuracy, but higher degree of accuracy does not necessarily represent that the
method is precise.

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 SIGNIFICANT FIGURES

Significant figures are digits which are necessary to express the results. Accuracy
of a measurement is depending upon the number if significant figures used to
represent it. More the significant figures more will be the accuracy.

For example, a sample weighed on rough balance is 71.2 and 71.2932 in analytical
balance. Here in rough balance measurement, there are only three significant
figures while in analytical balance there are six and each value give the precision
of respective measurement.

Rules for significant figures:

1. All non-zero numbers are significant.

e.g., 1,2,3,4,5,6,7,8,9

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2. Zeros between non-zero numbers are significant.
e.g., 00340.03210
3. Leading zeros are NEVER significant.
e.g., 00340.03210
4. Zeros on the end of a number are significant if there is a decimal.
e.g., 00340.003210

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