Bcm Instrumentation Flashcards
Who invented chromatography
Mikhail tsvet
Basic principles of chromatography
Differential distribution between the stationary n mobile phase
What is the stationary phase for paper chromatography
Cellulose acetate
Principle of differential distribution
The flow rate of the mobile phase
Retardation brought by the interaction of the solute n stationary phase
The speed of migration of solute depends on
Adsorptive affinity
In adsorption mechanism
Solute is separated base on
Adsorptive affinity
In partition mechanism solute is separated base on
Solubility difference
In exclusive mechanism solute are separated
By pore size
The movement of solvent in paper chromatography is through
Capillary action
The rate of mobility in paper chromatography is affected by
Relative solubility
Polarity of solvent
Polarity of solute
Reagent for paper chromatography
10%sulphuric acid exposed to iodine vapour
Thin layer chromatography make use of
Finely powered adsorbent such as :silica gel alumina starch biogel
Basic principles for separation of gel filtration is
Molecular size
Which separation technique s there no interaction between the solute molecules n stationary phase
Gel filtration
Basic principles for ion exchange chromatography
Charge to charge interaction
The stationary phase of iec consist of fixed charges know an
Resins
Basic principle of gas chromatography
Difference in the partition of the various molecules between the two phase
Basic principle for hplc
Separation based on relative affinity
What is the bases of hplc separation
Distribution of analyte between the mobile n stationary phase
The study of electro magnetic radiation emitted or absorbed by a given chemistal species
Spectroscopy
The duration of sample spent on column in hplc is determined by
Intermolecular interaction between it molecules and it parking material
What is used for measurements in spectrometer
A photo electric cell
What isolate radiant energy and narrow wave length region
A prism
What compares the light intensity between ntwo light path one path containing ba reference standard and the other a test sample
A double beam spectrometer
What measure the light intensity before and after a test sample has been inserted
Single beam spectrometer
The visible region is between
400_700nm
Ultra violent region of spectrum is between
400-200nm
Infra red region of spectrum is between
700-2000nm
Protein in solution have absorbance Maxima of 280nm because of
Because of amino acid with aromatic rings
Nucleic acid have an absorbance Maxima of 260nm
Because of the presence of heterocyclic rings
Photo metric instrument that measure visible region of spectrum is
Colorimeter
Fraction of light absorbed is
Absorbance
Fraction of light transmitted is
Transmittance
———-state that the absorption of radiant energy is directly proportional to the absorbing species in the sample
Beers law
Basic relationship between conc of absorbing species and measured value of absorbed radiation is in which law
Beers law
—————states that the same proportion of incident light will absorbed a unit thickness irrespective of the intensity of light
Lambert law
In beer Lambert law A=£cl
What is £
Molar extinction
—————-states the amount of light transmitted decrease exponentially with an increase in concentration of the solution through which the light passes
Beer Lambert law
According to beer Lambert law absorbance is linearly proportional to concentration but only valid for
Low concentration
According to beer Lambert high concentration might lead to
Association of molecules
________measures the density of light absorbed or transmitted
Spectrum photometry
Molecules that are excited by absorbtion of radiation are known as
Chromophore
If light is absorbed the molecules is
Excited
Absorbtion of longer wavelength such as ultraviolet n visible light is restricted to
Limited number of chromophore
What determine the absorption property of chromophore
Chemical structure of the molecule
Basic principles of centrifugation technique
Separation of particles based on sedimentation properties
Centrifugal equipment used in centrifugation technique
Centrifuge
The dense particle of centrifugation is termed
Pellet
The less dense part of centrifugation is termed
Supernatant
There are two procedure of centrifugation
Preparative
Analytical
______involvea the study of sedimentation characteristics of biological macro molecules
Analytical technique
______involves actual separation isolation and purification of particles such as whole cell an sub cellular organelles
Preparative technique
What respond to changes in hydrogen ion concentration
Ph electrode
What provide a stable electrical potential
Reference electrode
An increase in acidity of solution leads to
Higher concentration of hydrogen ion
An increase in alkalinity leads to an
Increase in hydroxyl ions
Basic principles of pH meter or potentiometer is based on
Exchange of ions between sample solution and inner solution of electrode
The unit of absorbance
Mol-1 cm-1
The force required to displaced solvent as the particle move through the centrifugal field
Bouyant force
Distribution is quantified using which coefficient
Distribution of partition coefficient
Sedimentation of particles is dependent on
Relative centrifugal force
Sedimentation coefficient is expressed in
Sverdberg unit
_______is a stepwise procedure involving increase centrifugal speed and time
Differential Centrifugation
An increase in s value will cause
Particle to sediment fast
Centrifugation through dense medium is
Density gradient centrifugation
Medium used in density gradient centrifugation
Sucrose
Fivoll
Precoll
Caesuim chloride
Two types of density gradient centrifugation are
Rate zonal
Isopycnic
The ratio of centrifugal force to the force of gravity is
Relative centrifugal force
Isopycnic centrifugation is used to separate
Organelles such as Golgi apparatus
Mitochondria perisome
In which centrifugation is the density of the particles to be separated are greater than the particles of the solvent
Rate zonal density
Which gradient marker produce continuous gradient
Sucrose
Basic principles of separation of rate zonal density centrifugation
Separation is based on size
Rf value is found in which chromatography
Paper chromatography
Positively resins are known as
Anion exchange
Beer Lambert states that intensity of light increase in respect to
Concentration of species
Mathematical expression for absorbance
—-logT. Log1/T logIo/I
Centrifugation depends on which factors
Viscosity
Size
Density
Shape
Which technique is used to detecting and quantifying analyte within a biological sample
Tracer technique
Which method is used in detecting an d quantifying analyte within biological sample based on specific antigen to antibody reaction
Immunoassay
Components of immunoassay are
Target analyte
Antibody
Detection label
Immune system are made up of
Humoral & cellular
ELISA means
Enzymes linked immunosbent assay