Pharmaceutical

Analysis
Definition and scope
Sem I
Pharmaceutical Analysis
 Different
techniques
of
analysis
 Analytical chemistry
• Application of a process or series of processes in
order to identify and/or quantify a substance, the
components of a solution or mixture, or the
determination of the structure of chemical
compounds.
• The “analytical technique” is a method that is used to
determine a chemical or physical property of a chemical
substance, chemical element, or mixture.

• There are a wide variety of techniques used for analysis,

from simple weighing (gravimetric analysis) to titrations
(titrimetric) to very advanced techniques using highly
specialized instrumentation.
 Types on basis of analyte size
• Macro Analysis-0.1g
• Meso (Semimicro) Analysis-0.01 to 0.1g
• Micro Analysis-0.001g to 0.01g
• Submicro Analysis-0.0001g to 0.001g
• Ultromicro Analysis-0.0001g
Classification of Analytical Chemistry
Qualitative Analysis
• It gives information about atomic and molecular
species or the functional group in sample.
• Melting point, boiling point, specific gravity,
Refractive Index, Absorbance, Optical rotation,
Viscosity, particle size, etc.
• Quantitative Analysis
• It gives relative amount of one or more of the
analyte in numerical terms.
 Analytical Methods
• Chemical Methods
• Physico-chemical Methods
• Microbiological Methods
• Biological Methods
• Chromatographic techniques
Chemical Methods
 Volumetric analysis
• Volumetric analysis, any method of quantitative
chemical analysis in which the amount of a
substance is determined by measuring the volume
that it occupies or, in broader usage, the volume of
a second substance that combines with the first in
known proportions, more correctly called
titrimetric analysis
 Gravimetric analysis

• Gravimetric
analysis describes a set
of methods used in
analytical chemistry for
the
quantitative determinati
on of an analyte (the ion
being analyzed) based
on its mass.
 Gasometric analysis
The determination of the structure or quantity of a substance by
means of measuring its gaseous derivatives.
 Physico-chemical Methods
Physical properties Instrumental methods
Electric potential Potentiometry
Electric conductance Conductometry
Electric current Polarography
Voltametry

Absorption of radiation Spectrophotometry

Colorimetry
Atomic absorption spectroscopy

Emission of radiation Emission spectroscopy

Flame photometry
Fluorimetry

Scattering of radiation Turbidimetric

Nephelometry

Refraction of radiation Refractometry

 Physical properties Instrumental methods

Rotation of plane polarised light Polarimetry

Optical rotatory dispersion

Thermal properties Thermal method

Mass to charge ratio Mass spectroscopy

 Potentiometry

• Potentiometry is one of the methods of electroanalytical

chemistry. It is usually employed to find the concentration of
a solute in solution. In potentiometric measurements, the
potential between two electrodes is measured using a high
impedance voltmeter.
 Conductometry
• The principle of conductometric titration is based on the fact
that during the titration, one of the ions is replaced by the
other and invariably these two ions differ in the ionic
conductivity with the result that conductivity of
the solution varies during the course of titration.
Polarography

• Polarography is a type of
voltammetry where the
working electrode is a
dropping mercury electrode
(DME) or a static mercury
drop electrode (SMDE), which
are useful for their wide
cathodic ranges and
renewable surfaces.
 Voltametry
• Voltammetry is a category of electroanalytical methods used
in analytical chemistry and various industrial processes.
In voltammetry, information about an analyte is obtained by
measuring the current as the potential is varied.
 Spectrophotometry

• Spectrophotometry is a method to measure how much a

chemical substance absorbs light by measuring the intensity of
light as a beam of light passes through sample solution. The
basic principle is that each compound absorbs or transmits light
over a certain range of wavelength.
 Colorimetry
• In physical and analytical chemistry, colorimetry or colourimetry
is a technique used to determine the concentration of colored
compounds in solution. A colorimeter is a device used to test
the concentration of a solution by measuring its absorbance of a
specific wavelength of light.
 Atomic absorption spectroscopy

• Atomic absorption spectrometry (AAS) detects elements in

either liquid or solid samples through the application of
characteristic wavelengths of electromagnetic radiation from a
light source.
• Depending on the light wavelength and its intensity, specific
elements can be detected and their concentrations measured.
 Emission spectroscopy

• Emission spectroscopy is
a spectroscopic technique which examines the
wavelengths of photons emitted by atoms or
molecules during their transition from an
excited state to a lower energy state.
 Flame photometry

• flame photometry, a branch

of atomic spectroscopy is
used for inorganic chemical
analysis for determining the
concentration of certain
metal ions such as sodium,
potassium, lithium, calcium,
Cesium, etc. In flame
photometry the species
(metal ions) used in the
spectrum are in the form of
atoms.
 Fluorimetry

• A fluorometer or fluorimeter is a device used to measure

parameters of visible spectrum fluorescence: its intensity and
wavelength distribution of emission spectrum after excitation by
a certain spectrum of light. These parameters are used to identify
the presence and the amount of specific molecules in a medium.
 Turbidimetric

• Turbidimetry (the name being derived from turbidity) is the

process of measuring the loss of intensity of transmitted light
due to the scattering effect of particles suspended in it. Light is
passed through a filter creating a light of known wavelength
which is then passed through a cuvette containing a solution.
 Nephelometry

• Nephelometry is a technique used in immunology to determine

the levels of several blood plasma proteins. For example the
total levels of antibodies isotypes or classes: Immunoglobulin
M, Immunoglobulin G, and Immunoglobulin A. It is important in
quantification of free light chains in diseases such as multiple
myeloma.
 Refractometry
• Refractometry is the analytical method of measuring substances'
refractive index (one of their fundamental physical properties) in
order to, for example, assess their composition or purity. A
refractometer is the instrument used to measure refractive index
("RI").
 Polarimetry

• Polarimetry is the measurement and

interpretation of the polarization of transverse
waves, most notably electromagnetic waves,
such as radio or light waves.
 Optical rotatory dispersion

Optical rotatory dispersion is

the variation in
the optical rotation of a
substance with a change in the
wavelength of light.
For example, when plane-
polarized white light from an
overhead projector is passed
through a cylinder of sucrose
solution, a spiral rainbow is
observed perpendicular to the
cylinder.
 Thermal method
• Thermal methods are multi-component techniques and include
thermogravimetry, differential thermal analysis and differential
scanning calorimetry.
• These methods find widespread use in both quality control and
research applications on industrial products such as polymers,
pharmaceuticals, metals and alloys.
 Mass spectroscopy

Mass spectrometry (MS) is an analytical technique that measures

the mass-to-charge ratio of ions. The results are typically presented
as a mass spectrum, a plot of intensity as a function of the mass-to-
charge ratio.
 Microbiological Methods

• Cylindrical Plate (or Cup Plate) Method

• Turbidimetric (or Tube assay) Method
Cylindrical Plate (or Cup Plate)
Method
Turbidimetric (or Tube assay) Method
 Biological Methods

• Matching Point or Bracketing Method

 Semi quantitative method
• Limit test - defined as quantitative or semi
quantitative test designed to identify and control small
quantities of impurity which is likely to be present in the
substance. Limit test is generally carried out to determine
the inorganic impurities present in compound.
 Chromatographic techniques
• Paper chromatography
• Ion exchange chromatography
• Affinity chromatography
• Thin layer chromatography
• High performance liquid chromatography
• Gas chromatography
• High performance thin layer chromatography
 Paper chromatography

• The principle involved is partition chromatography wherein the

substances are distributed or partitioned between liquid phases.
One phase is the water, which is held in the pores of the
filter paper used; and other is the mobile phase which moves
over the paper.
 Ion exchange chromatography

• Ion-exchange chromatography separates molecules based on

their respective charged groups.
• Ion-exchange chromatography retains analyte molecules on the
column based on coulombic (ionic) interactions.
• The ion exchange chromatography matrix consists of positively
and negatively charged ions.
 Affinity chromatography

• Affinity chromatography is a method of separating a biomolecule

from a mixture, based on a highly specific macromolecular
binding interaction between the biomolecule and another
substance.
 Thin layer chromatography
• Thin-layer chromatography is a chromatography technique used
to separate non-volatile mixtures.
• Thin-layer chromatography is performed on a sheet of an inert
substrate such as glass, plastic, or aluminium foil, which is coated
with a thin layer of adsorbent material, usually silica gel,
aluminium oxide, or cellulose.
 High performance liquid
chromatography
• HPLC relies on pumps to pass a pressurized liquid and a sample
mixture through a column filled with adsorbent, leading to the
separation of the sample components.
• The active component of the column, the adsorbent, is typically
a granular material made of solid particles (e.g., silica, polymers,
etc.), 2–50 μm in size.
 Gas chromatography

• Gas chromatography (GC) is an analytical technique used to

separate the chemical components of a sample mixture and
then detect them to determine their presence or absence
and/or how much is present. These chemical components are
usually organic molecules or gases.
 High performance thin layer
chromatography
• H.P.T.L.C. is very useful qualitative analysis method; it combines
the art of chromatography with quickness at a moderate cost.
• It is a major advancement of TLC principle with short time
duration and better resolution. gel of very fine particle size is
widely used as adsorbent.
 Importance of Pharmaceutical
Analysis

• Identification of raw materials (in-process and

finished product).
• Determination of additional impurities.
• Stability of the drug.
• Strength and concentration of the chemical
compound.
• Determine molecular weight of the chemical
compound.