OVERVIEW when equilibrium is reached. PRINCIPLE: The porous membrane

OVERVIEW
OF BIOCHEMICAL TECHNIQUES

INTRODUCTION:

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Biochemical
techniques refer to a set of assays, procedure and approaches that helps to
researcher  in order to identify or study
 the materials found inside the  organisms and its  chemical reactions.

BIOCHEMISTRY RELATED TO APPLIED
DISCIPLANE:

§  Biochemistry
is mixed with both clinical and applied discipline that involves in is both
analytical & quantitative methods in order 
to explain the impact of relationship in molecular level.

§  In
biochemical research, experimental models are first subjected to qualitative
evaluation i.e., the complex device is damaged down & components are
separated & identified.

APPLICATION OF BIOCHEMICAL TECHNIQUES

§  It
gives an opportunity for researchers and scientists to discover the modern
research techniques within the discipline of biochemistry .

§  It
could be used each for separation & identity of macromolecules & micro
molecules.  In biochemistry, the term
macromolecule is applied to the four biopolymers ( nucleic acids, proteins ,
carbohydrates & lipids ) in addition to non-polymeric molecules with high
weight.

BIOCHEMICAL TECHNIQUES

1.     
DIALYSIS

2.     
CHROMATOGRAPHY

3.     
ELECTROPHORESIS

4.     
SPECTROPHOTOMETRY

5.     
MASS SPECTROSCOPY:

6.     
CENTRIFUGATION

1.   DIALYSIS:

This
method involves small solutes diffuse from a low concentration solution to high
conc. solution across a semi permeable membrane when equilibrium is reached.

PRINCIPLE:

The porous membrane
selectively  pass small solute  with  maintaining of large solute in solution . it
is separation process  of molecules primarily
based on size .

2.   CHROMATOGRAPHY:

A
technique which is used in the separation of mixture is known as
chromatographic technique. Chromatography technique is a laboratory technique.

Chromatography is an analytical
approach dealing with the separation of closely related compound from a
aggregate mixture . These include protein, peptides, amino acid, lipids, carbohydrates,
vitamins .

PRINCIPLE:

§  Mobile phase: A
solvent that flows through the supporting medium

§  Stationary phase: A
layer and the coating of the supporting medium that interact with the analyte

§  Supporting medium is a
solid surface on which the stationary phase is bounded or coated.

SIGNIFICANCE

The importance of the
chromatography is versatility and small quantities of a material can be
separated with case it is fast and accurate.

TYPES OF 
CHROMATOGRAPHY

§  PAPER CHROMATOGRAPHY

It is  a beneficial method in oreder to seprate the
combination of amino acid, sugar, chemical substances, lipid, urea in solution.
 Separation is primarily based on a
liquid-liquid partition of the compounds.

§  HIGH PERFORMANCE LIQUID CHROMATOGRAPHY
(HPLC)

It is used to separate  elements from complex solution  by the use of chemical interactions between
the materials being analyzed & the chromatography column.

3.   ELECTROPHORESIS:

The motion of charged ions
under the effect of electrical field  that results in migration towards the
oppositely charged electrode is called electrophoresis .OR

It is the motion of dispersed particle
relative to a fluid under the influence of spatially uniform electric field

SIGNIFICANCE

Importance in the separation of
macromolecules such DNA, RNA and protein

TYPES OF ELECTROPHORESIS

§  PAPER ELECTROPHORESIS

 Separation of charged particles is determined
by using differences in their migration size which varies with electric charge
, length of particle & form of particles.

§  GEL ELECTROPHORESIS,

It is used for the separation
of proteins & nucleic acid.  This technique
helps for  separation of molecules based
on their size and charge.

§  ISOELECTRIC FOCUSSING.

Basically used for purification
of proteins.  This technique is primarily
based on the immobilization of the molecules at isoelectric pH at some point of
electrophoresis.

4.   SPECTROPHOTOMETRY

Spectrophotometry
is commonly used for the measurement of transmittance of reflectance of
solution, transparent or opaqness in sample.

It is a method to measure how
much a chemical absorbs light, by measuring the intensity of light as a beam of
light passes through sample solution.

 Also it gives the quantitative dimension of
the mirrored image molecules.

SIGNIFICANCE

Its importance in medical
research, development of medicine research on vitamin and hormones etc.

5.   MASS
SPECTROSCOPY

It is analytical techniques in
which chemical species are ionized and these ions are sorted based on their
mass to charge ratio

 SIGNIFICANCE

Mass spectroscopy is used in
pharmaceutical, clinical, environmental, Biotechnology

 

6.   CENTRIFUGATION

Is a device for separating   particles from a solution according to there
length, shape, viscosity, density of the medium.

Centrifugation is
a process in which the centrifugal force are applied for the sedimentation of
the heterogeneous mixture with a centrifuge

SIGNIFICANCE

Used for the
separation of molecules on the basis of density and used in industries research