Professional Documents
Culture Documents
Objectives
To separate the components of Capsicum frutescens / Moringa
oleifera
To perform column chromatography and thin layer chromatography
To identify the colored pigments present in Capsicum frutescens /
Moringa oleifera
To determine the polarity of the colored pigments
To compute the Rf value of the colored pigments
Plant Sources
Capsicum frutescens
Moringa oleifera
Chromatography
separation of mixtures due to differences in the
equilibrium distribution of sample components
between stationary phase and mobile phase.
separation is based on the rates at which the
components of a mixture are carried through a
stationary phase by a mobile phase
Mobile Phase
Moves over or though the stationary phase
carrying the pigments in it
organic
compounds
interact
with
the
adsorbents through the following types of
interactions: ion-dipole, dipole-dipole, hydrogen
bonding, dipole induced dipole, and van der
Waals forces
Silica and Alumina
ELUTION
Chromatogram
pattern formed by substances that have been
separated by chromatography
output or
technique
result
of
the
chromatographic
Types of Chromatography
Based on direction of movement of the solvent
1. ascending (through capillary action)
2. descending (due to action of gravity)
1. Isocratic
2. Gradient
Liquid
STATIONARY PHASE
TYPE
Solid; adsorbent
Gas-solid (adsorption)
Gas-solid
Liquid
Gas-liquid (partition)
Solid; adsorbent
Liquid-Solid (adsorption)
Solid; gel
Solid; immobilized
Liquid
Liquid-liquid (partition)
Liquid
STATIONARY PHASE
TYPE
Paper
Paper (partition)
Sample Preparation
Column Chromatography
one of the most useful methods for
the separation and purification of
solids and liquids when purifying
small quantities of material
adsorbent
cotton
plug
Column Chromatography
Elution with the Solvent System
solvent
adsorbent
(1) the
(2) the
Column Chromatography
Development of Chromatogram
Mr. A. R. M. Salcedo
Ethers
Increasing polarity
Halogenated hydrocarbons
Aromatic hydrocarbons
Alcohols
Amines
Carboxylic acids
Mr. A. R. M. Salcedo
Column Chromatography
Components of Capsicum frutescens
Stationary phase: silica gel
Column Chromatography
Components of Capsicum frutescens
Column Chromatography
Components of Capsicum frutescens
CAPSANTHIN (red)
CAPSORBIN (red-orange)
Column Chromatography
Components of Capsicum Frutescens
-CAROTENE (yellow)
Column Chromatography
Components of Capsicum frutescens
Column Chromatography
Components of Moringa oleifera
Stationary phase: silica gel
Mobile phase:
1. Hexane:acetone (7:3)
2. Acetone
3. Acetone: methanol (1:1)
Column Chromatography
Components of Moringa oleifera
lutein
zeaxanthine
Column Chromatography
Components of Moringa oleifera
Chlorophyll a (dark green)
Column Chromatography
Components of Moringa oleifera
Phaeophytin a & b
(dark gray and bluish gray)
adsorbent
spots of sample
origin
2. Equilibration
3. Development of Chromatogram
4. Visualization
5. Evaluation
Rf =
Ultraviolet Spectroscopy
- used to detect the presence of a conjugated pi bond
system in a molecule
The Electromagnetic
Spectrum
Ultraviolet Spectroscopy
Observed Color
of Compound
Approximate Wavelength
of Light Absorbed
Green
Red
700 nm
Blue-green
Orange-red
600 nm
Violet
Yellow
550 nm
Red-Violet
Yellow-green
530 nm
Red
Blue-green
500 nm
Orange
Blue
450 nm
Yellow
Violet
400 nm
The End
Miracles are reserved to those
who believe according to their
faith.