Different Techniques

In Pharmaceutical Analysis

Hafizul Seikh
Bengal College of Pharmaceutical Sciences and Research

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1.2 TECHNIQUES IN PHARMACEUTICAL ANALYSiS
There are various techniques of pharmaceutical analysis which can be divided into two
major categories.
1. Qualitative analysis
2. Quantitative analysis
1. Qualitative Analysis:
This category of analysis involves various test procedures that are designed for the
identification of compounds in the sample. These test results confirm the presence or
absence of a compound in the sample to be analysed.
Example: Colour reaction tests, limit tests, melting point and boiling point determination
for identification, precipitate formation etc.
2. Quantitative Analysis:
This category of analysis involves the quantitative determination of compounds in the
sample. Generally, the quantitation has been done on the basis of some physical property of
components. Quantitative analytical techniques are further classified as follows:
1. Chemical Methods
(a) Volumetric, (b) Gravimetric, (c) Gasometric
2. Physico-chemical Methods or Instrumental Methods
3. Microbiological Methods
4. Biological Methods
1.2.1 Chemical Methods
(a) Volumetric Methods: In volumetric methods, measurement of volume of solution is
taken as a parameter for assay. The volume of known strength of a solution that is required
to react completely with the substance to be analyzed is measured. The quantity of analyte is
determined from the volume of solution by calculation. The solution or reagent is called as
titrant and the analyte to be analysed is termed as titrate. The ideal characteristics in this
method to be considered are that, the reagents selected should react rapidly with the
analyte, the reaction must be complete and there should be some method of detection to
detect the end of reaction. Volumetric or titrimetric methods are rapid and user friendly
compared to gravimetric methods, thus preferred. Volumetric methods are classified into
different types depending upon the type of reactions involved in the reaction which are as
follows:
Neutralization titrations 2. Non-aqueous titrations
3. Precipitation titrations 4. Oxidation-reduction titrations
5. Complex metric titrations.

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 (b) Gravimetric Methods: In Gravimetric analysis, quantitation is done on the basis of
weight of compound. This process involves isolation and weighing of the compound of
known composition, i.e. purest form. The analysis is carried out by various processes such as
precipitation, volatilization, electro-analytical etc. Precipitation and volatilization are widely
employed methods. Gravimetric analysis is time-consuming compared to other techniques. It
is useful for the examination of compounds for the presence of impurities.
(c)Gasometrical Methods: In this method, the measurement of the volume of gases
forms the basics of analysis. When a chemical reaction is carried out under the specific
process, the volume of gas evolved or absorbed in the reaction is measured. The measured
volume is corrected to standard conditions of temperature and pressure. Gas burettes or
nitro-meters are used for the measurement of volume of gas. Example, for gases that are
measured by gasometrical methods are cyclopropane, carbon dioxide, nitrous oxide, oxygen,
octal nitrate, nitrogen, amyl nitrate, ethylene and helium.
1.2.2 Instrumental Methods
These methods involves the usage of instruments to measure the physical or
physiochemical property of the compound to be analysed thus lead to quantitaion of the
compound. Depending on the physical property of the compound various instruments have
been used for the measurement. The measurement of change in properties such as electric
current, potential, electrical conductivity, optical density, scattering of radiation, absorption,
emission, refractive index etc. using suitable, reliable and sensitive instruments. Some of the
physical properties and corresponding equipment used are listed as follows.
Physical Properties Instrumental Methods
Electrical potential Potentiometer
2. Electrical conductance Conductometry
3. Electrical current Polarography and voltametry
4. Absorption of radiation Spectrophotometry,
Colorimetry
Atomic absorption spectroscopy
5. Emission of radiation Emission spectroscopPy
Flame photometry,
Fluorimetry
6. Scattering of radiation Turbidimetric and Nephelometry
7. Refraction of radiation Refractometry
Rotation of plane Polarimetry
polarized light Optical rotatory dispersion
9. Thermal properties Thermal method of analysis
(DSC, DTA, TGA)

10. Mass to charge ratio Mass spectrometry

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 Further, instrumental methods involve another major area of analysis for separation
purpose called as chromatography. Chromatographic techniques are the separation
techniques used to separate components from a mixture of two or more compounds. They
use generally a stationary phase and a mobile phase for separation using adsorption and
partition as the major principles. Various techniques are involved in chromatography
including column chromatography, HPLC, thin layer chromatography, HPTLC, paper
chromatography, gel chromatography, gas chromatography, flash chromatography, ion
exchange chromatography and ion pair chromatography. These techniques have extensive
applications in the pharma industries.
1.2.3 Microbiological Methods
Microbiological methods are employed for compounds especially for antibiotics, for
which the chemical methods are not useful. This method involves the determination of
inhibition of growth of bacteria by the substances to be analysed in comparison with the
standard compound. On the basis of the result, the therapeutic efficacy of the antibiotics are
decided. The methods which are generally used include the cylinder plate (or cup plate)
method and the Turbidimetric (or tube assay) method.
1.2.4 Biological Methods
Biological assays are carried out to observe the biological effect of the drug on some
type of living matter. They are also called as bioassays. These are recommended when the
chemical or physical methods are not capable to estimate the potency of a drug. These
methods are carried out by comparing the biological effect of the sample to be tested with
the biological effect produced by a standard compound in identical test conditions. However,
the results of biological assays vary due to the difficulties in maintaining the identical
conditions thus the errors are also to be considered and corrected. Bioassays involves
measurement of various parameters including weight of tissues of organ, weight of organs,
blood parameters such as blood glucose, cholesterol, urea, enzyme concentration etc. In
some assays even the physiological symptoms also have been considered.

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