Separation techniques play a key role in various fields of science and technology. The document discusses the significance of separation techniques in pharmaceutical analysis. It covers various classical and instrumental separation methods like chromatography, spectroscopy, centrifugation etc. and their applications in qualitative analysis, quantitative analysis and purification of substances. The conclusion emphasizes that separation techniques are important as they allow obtaining pure materials from complex mixtures.

1. A Seminar On Significance Of Separation
Techniques In Pharmaceutical Analysis
Presented By, Guided By,
Miss. Madhavi Mutha Dr. Rajesh J Oswal
Prof.Sandip Kshirsagar
Department of Pharmaceutical Chemistry
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2. INTRODUCTION
. Separations touch every branch of
science and technology and have
developed into a well established branch
known as separation science. If you
just look at the developments, say in
biological sciences such as
biochemistry, biotechnology, clinical
pharmacology, therapeutics and
toxicology, the progress has taken place
prominently because of the
advancements in the separation
methods.
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3. SEPARATION METHODS − A UNIFIED
SCIENCE
 The separations play a key role in the various branches
of science and technology but they themselves form a
unified branch of science. In the simplest terms,
separation is defined as an operation in which a
mixture is divided into at least two components having
different compositions. But this particular definition has
a limitation as it does not cover chiral separations in
which molecules of same composition and chemical
structure are involved. Therefore, a broader definition
of separation will be as:“Separation is a process by
which a mixture is divided in at least two
components with different compositions or two
types of molecules with the same composition but
different stereochemistry.”
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4. Objectives
i) Analysis of different constituents of a mixture.
ii) Procuring pure materials from complex mixtures.
In analysis, (i), there may be two aspects:
• Removal of interfering constituents before the actual
quantitation of one or more known compounds.
• Isolation of unknown constituents for subsequent
characterization.
Under “(ii)”, for obtaining the pure materials from
complex mixtures, the constituent with the desired purity.
In some cases, in order to attain the required level of
purity, the separation steps of the same process may
have to be repeated.
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5. SIGNIFICANCE OF SEPARATION TECHNIQUES
Analytical chemistry:
1.Classical methods:
. Qualitative analysis
A qualitative analysis determines the presence or absence of a particular
compound, but not the mass or concentration.
.Flame test
.Inorganic qualitative analysis generally refers to a systematic scheme to
confirm the presence of certain, usually aqueous, ions or elements by
performing a series of reactions that eliminate ranges of possibilities and then
confirms suspected ions with a confirming test..
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6. Gravimetric analysis:
Gravimetric analysis involves determining the amount of material
present by weighing the sample before and/or after some
transformation. A common example used in undergraduate education
is the determination of the amount of water in a hydrate by heating the
sample to remove the water such that the difference in weight is due
to the loss of water.
.Volumetric analysis:
Titration involves the addition of a reactant to a solution being
analyzed until some equivalence point is reached. Often the amount
of material in the solution being analyzed may be determined.
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7. 2.Instrumental methods:
.Spectroscopy:
Spectroscopy measures the interaction of the molecules with
electromagnetic radiation. Spectroscopy consists of many
different applications such as atomic absorption spectroscopy,
atomic emission spectroscopy, ultraviolet-visible spectroscopy,
x-ray fluorescence spectroscopy, infrared spectroscopy, Raman
spectroscopy, dual polarisation interferometry, nuclear magnetic
resonance spectroscopy, photoemission spectroscopy,
Mössbauer spectroscopy and so on
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8. .Mass spectrometry :
Mass spectrometry measures mass-to-charge ratio of molecules
using electric and magnetic fields
.Electrochemical analysis:
Genomics - DNA sequencing and its related research. Genetic
fingerprinting and DNA microarray
Proteomics - the analysis of protein concentrations and
modifications, especially in response to various stressors,at various
developmental stages, or in various parts of the body.
Metabolomics - similar to proteomics, but dealing with metabolites.
Transcriptomics - mRNA and its associated field
Lipidomics - lipids and its associated field
Peptidomics - peptides and its associated field
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9. .Thermal analysis:
Calorimetry and thermogravimetric analysis measure the
interaction of a material and heat.
.Separation:
Adsorption, adhesion of atoms, ions or molecules of gas, liquid,
or dissolved solids to a surface.
.Centrifugation:
Separating textile.
Removing water from lettuce after washing it in a salad
spinner.
Separating particles from an air-flow using cyclonic
separation.
The clarification and stabilization of wine.
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10. Chromatography:
Column chromatography :
Column chromatography is a separation technique in which the
stationary bed is within a tube. The particles of the solid stationary
phase or the support coated with a liquid stationary phase may fill the
whole inside volume of the tube (packed column) or be concentrated
on or along the inside tube wall leaving an open, unrestricted path for
the mobile phase in the middle part of the tube (open tubular column).
Differences in rates of movement through the medium are calculated
to different retention times of the sample.
Paper chromatography:
.separation and identification of mixtures that are or can be
colored, especially pigments.
Two-way paper chromatography, involves using two solvents and
rotating the paper 90° in between. This is useful for separating
complex mixtures of similar compounds, for example, amino acids.
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11. .Thin layer chromatography:
.analyzing ceramides and fatty acids.
.detection of pesticides or insecticides in food and water.
.analyzing the dye composition of fibers in forensics.
.assaying the radiochemical purity of
radiopharmaceuticals.
.Displacement chromatography:
.purification of proteins from complex mixtures.
.Two-dimensional chromatography represents rigorous approach to
evaluation of the proteome.
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12. .Gas chromatography:
 Gas chromatography (GC), is used in analysing
compounds that can be vaporized without decomposition.
It is well suited for use in the petrochemical, environmental
monitoring and remediation, and industrial chemical fields.
.Liquid chromatography:
High-performance liquid chromatography:
HPLC, is a chromatographic technique that can separate a
mixture of compounds and is used in biochemistry and analytical
chemistry to identify, quantify and purify the individual
components of the mixture.
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13. .Affinity chromatography:
.Purify and concentrate a substance from a mixture into a buffering
solution
.Reduce the amount of a substance in a mixture
.Discern what biological compounds bind to a particular substance
.Purify and concentrate an enzyme solution.
.Supercritical fluid chromatography:
(SFC) is a form of normal phase chromatography that is used for
the analysis and purification of low to moderate molecular weight,
thermally labile molecules. It can also be used for the separation of
chiral compounds
.Ion-exchange chromatography:
It has significance in water purification, preconcentration of trace
components, ligand-exchange chromatography, ion-exchange
chromatography of proteins, high-pH anion-exchange
chromatography of carbohydrates and oligosaccharides, and others.
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14. .Size-exclusion chromatography:
The main application of gel-filtration chromatography is the
fractionation of proteins and other water-soluble polymers, while gel
permeation chromatography is used to analyze the molecular weight
distribution of organic-soluble polymers.
Hybrid techniques
Gas chromatography–mass spectrometry:
GC-MS include drug detection, fire investigation, environmental analysis,
explosives investigation, and identification of unknown samples.
GC-MS has been widely heralded as a "gold standard" for forensic
substance identification because it is used to perform a specific test
Nuclear magnetic resonance spectroscopy:
NMR spectroscopy is often the only way to obtain high resolution
information on partially or wholly unstructured proteins.
"Nucleic acid NMR" is the use of NMR spectroscopy to obtain
information about the structure and dynamics of polynucleic acids,
such as DNA or RNA.
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15. Infrared spectroscopy:
Used in the long-term unattended measurement of
CO2concentrations in greenhouses and growth chambers by
infrared gas analyzers.
It also has significance in forensic analysis Infrared
spectroscopy has also been successfully utilized in the field
of semiconductor microelectronics.
Liquid chromatography–mass spectrometry:
LC-MS is very commonly used in pharmacokinetic studies of
pharmaceuticals and is most frequently used technique in the
field of bioanalysis and Metabolics.
LC-MS is frequently used in drug development at many different
stages including Peptide Mapping, Glycoprotein Mapping,
Natural Products Dereplication, Bioaffinity Screening, In Vivo
Drug Screening, Metabolic Stability Screening, etc.
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16. CONCLUSION: :
 It deals with separation of mixtures to enhance
purity of substance.
 Separation techniques are important because
most substances are needed in the pure state.
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