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CONTENT
CHROMATOGRAPHY_______________________________________________________
TYPES OF CHROMATOGRAPHY___________________________________________
Column Chromatography_________________________________________________
1. STATIONARY PHASE:___________________________________________________________
Adsorbents:______________________________________________________________________
The Solvent_____________________________________________________________________
Analysis of Column Eluents____________________________________________________
PREPARATION OF THE COLUMN:__________________________________________________
PACKING OF THE COLUMN:_______________________________________________________
1. Dry PACKING TECHNIQUE:_____________________________________________________________
2. WET PACKING TECHNIQUE:___________________________________________________________
DETECTION OF COMPONENTS:____________________________________________________
FACTORS AFFECTING COLUMN EFFICIENCY:______________________________________
APPLICATIONS___________________________________________________________________
ADVANTAGES OF C.C.:____________________________________________________________
DISADVANTAGES OF C.C.:_________________________________________________________
Pharmacognosy
CHROMATOGRAPHY
TYPES OF CHROMATOGRAPHY
1. Solid liquid Chromatography
Stationary Phase----Solid, Mobile Phase------Liquid
Tech: C.C., TLC, FCC, HPLC
2. Liquid liquid Chromatography
Stationary Phase----Liquid, Mobile Phase------Liquid
Tech: Paper Chromatography
3. Gas - Liquid Chromatography
Gas - Solid Chromatography
Stationary Phase----Solid / liquid, Mobile Phase------Gas
Tech: Gas Chromatography
Column Chromatography
1. STATIONARY PHASE:
CRITERIA:
Spherical in shape
Mechanical stability must be high.
Pharmacognosy
They should not react chemically.
It should be useful for separating for wide variety of compounds.
It shud be freely available and inexpensive.
Adsorbents:
Typical adsorbents used in C.C. are Alumina (aluminum oxide), Activated
charcoal, Magnesium silicate, Silica gel (silicon dioxide)., Inorganic carbonates,
starch, cellulose, sucrose.
Silica gel is slightly acidic while alumina may be acidic, neutral or basic. For
greatest effectiveness the particles of solid adsorbent should be of uniform size
and large surface area.
The strength of adsorption depends upon the compounds involved. Since the
adsorbents are polar, the more polar compounds are adsorbed more strongly.
Thus, non-polar compounds are eluted first.
The Solvent
The polarity of the solvent which is passed through the column affects
the relative rates at which compounds move through the column.
Polar solvents can more effectively compete with the polar molecules
of a mixture for the polar sites on the adsorbent surface and will also
better solvate the polar constituents.
Pharmacognosy
Analysis of Column Eluents
DETECTION OF COMPONENTS:
Colored components----- Visually
Colourless components--------Different properties which can be used are
UV / Visible detector, Flourescence detector, RI detector, Flame Ionization
Detector,..
FACTORS AFFECTING COLUMN EFFICIENCY:
Nature of the solvent
Dimensions of columns
Particle size of column packing
Temperature of the column
Pharmacognosy
APPLICATIONS
Separation of mixture if the components
Purification process
Isolation of metabolites
Estimation of drugs in formulation.
ADVANTAGES OF C.C.:
Any type of mixture can be separated
Any quantity of mixture can be separated
Wider choice of M.P.
Automation is possible
DISADVANTAGES OF C.C.:
Time consuming
Increased amounts of M.P. is required.
Automation makes the technique more complicated and expensive.
PLANT PIGMENTS
The plant life around us is full of wonderfully rich and vibrant colors. From red
to violet, the entire color spectrum can be found in plant materials. These colors
come
from a surprisingly small number of plant pigments. The three primary groups of
plant
pigments are anthocyanins, carotenoids, and chlorophyll. Others include porphyrins,
betalains, flavonoids etc. Each of these absorbs light at
different parts of the visible light spectrum and has a different function in the plant.
The light that is absorbed may be used by the plant to power chemical
reactions, while the reflected wavelengths of light determine the color the
pigment will appear to the eye.
Pharmacognosy
Chlorophyll is the primary pigment in plants; it is a chlorin that absorbs yellow
and blue wavelengths of light while reflecting green. It is the presence and
relative abundance of chlorophyll that gives plants their green color. T he energy
of sunlight is captured by chlorophyll pigments to make food during the process of
photosynthesis.
There are 3 kinds of Chlorophyll:
Chlorophyll "a": mainly responsible for photosynthesis. Found in all plants, algae and
cyanobacteria carrying out photosynthesis.
Chlorophyll "c": found only in photosynthetic kind of organisms found in aquatic ecosystem
e.g. Chromista and dinoflagellates.
All chlorophylls serve as the primary means plants use to intercept light in order
to fuel photosynthesis.
Betalains are red or yellow pigments. Like anthocyanins they are water-soluble,
but unlike anthocyanins they are indole-derived compounds synthesized from
tyrosine. This class of pigments is found only in the Caryophyllales (including
cactus and amaranth), and never co-occur in plants with anthocyanins.
Pharmacognosy
Betalains are responsible for the deep red color of beets, and are used
commercially as food-coloring agents.
Nyctaginace
ae