ASSIGNMENT NO 1 Chromatography EXACT

ASSIGNMENT NO 1
CHROMATOGRAPHY:-
The word chromatography is derived from two Greek words Chroma means color and graphein means
to write.
Chromatography is a laboratory technique for the separation of a mixture. The mixture is dissolved in a
fluid called the mobile phase, which carries it through a structure holding another material called the
stationary phase.
 MOBILE PHASE:- It is the phase that moves in a definite direction for the chromatography
procedure.
 STATIONARY PHASE:- The stationary phase is the substance fixed in place for the
chromatography procedure.
TYPES OF CHROMATOGRAPHY:-
1. COLUMN CHROMATOGRAPHY:-
Column chromatography is a chromatography method used to isolate a single chemical
compound from a mixture. Chromatography is able to separate substances based on differential
adsorption of compounds to the adsorbent; compounds move through the column at different rates,
allowing them to be separated into fractions.
2. THIN-LAYER CHROMATOGRAPHY (TLC):-

Thin layer chromatography (TLC) is a chromatographic technique used to separate the
components of a mixture using a thin stationary phase supported by an inert backing.
3. GAS CHROMATOGRAPHY (GC):-

Gas chromatography (GC) is a common type of chromatography used in analytical chemistry for
separating and analyzing compounds that can be vaporized without decomposition.
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4. ION-EXCHANGE CHROMATOGRAPHY:-
Ion chromatography is a chromatography process that separates ions and polar molecules based
on their affinity to the Ion Exchanger. It works on almost any kind of charged molecule including
large proteins, small nucleotides, and amino acids.
5. PAPER CHROMATODRAPHY:-
Paper chromatography is one of the types of chromatography procedures which runs on a
piece of specialized paper. It is a planar chromatography system where in cellulose filter paper
acts as a stationary phase on which the separation of compounds occurs.
6. HIGH PRESSURE LIQUID CHROMATOGRAPHY (HPLC):-

High-performance liquid chromatography is a technique in analytical chemistry used to
separate, identify, and quantify each component in a mixture.
7. AFFINITY CHTOMATOGRAPHY:-
Affinity chromatography is a method of separating biochemical mixtures based on a highly
specific interaction between antigen and antibody, enzyme and substrate, receptor and ligand,
or protein and nucleic acid.
APPLICATIONS OF CHROMATOGRAPHY:-
 It is used in the food industries for analysis of different additives in the food.
 Environmental and governmental agencies also use chromatography to test drinking water.
 In pharmaceutical companies large amount of pure chemicals for making further medicines is
prepared by using chromatography.
 Paper chromatography and Gas chromatography are employed in finger print, DNA RNA
analysis.
 It is applied in forensic science for investigation.
 Presence of alcohol or some other drugs in blood or urine are detected by using gas
chromatography.
 It is also used to derive the amount of reactants or products.
PURPOSE OF CHROMATOGRAPHY:-
Chromatography consists of two phases: one mobile phases and one configuous stationary phase. The
stationary phase is liquid or solid and the mobile phase is gas or liquid. The compound mixture moves
along with the mobiIe phase through stationary phase and separate dispensing on the different degree
of adhesion (to the silica) of each component in the sample or the compound mixture

WHAT IS SILICA?
Silicon dioxide is also known as silica with the chemical formula SiO₂. It is a hard, unreactive, colorless
compound which occurs as the mineral quartz and as a principal constituent of sandstone and other
rocks.
WHAT IS N-HEXANE:-
Hexane is an alkane of six carbon atoms, with the chemical formula C₆H₁₄. n-Hexane is a chemical made
from crude oil. Pure n-Hexane is a colorless liquid with a slightly disagreeable odor. It is highly
flammable, and its vapors can be explosive. Pure n-Hexane is used in laboratories
WHAT IS THE RETENTION FACTOR (RF) ?
The Rf value is defined as the ratio of the distance moved by the solute (i.e. the dye or pigment under
test) and the distance moved by the solvent (known as the Solvent front) along the paper.
 If Rf value of a solution is zero,the solute remains in the stationary phase and thus it is
immobile.
 If Rf value=1 then the solute has no affinity for the stationary phase and travel with the
solvent front.
 Rf value can never be greater than 1.
USES CHROMATOGRAPHY:-
ANALYTICAL:- determine chemical composition of a sample.
PREPARATIVE:- purify and collect one or more components of a sample.
PIGMENTATION:- separate pigments due to difference in solubility.
CAPILLARY ACTION:- the movement of liquid within the spaces of a porous material due to the force of
adhesion, cohesion, and surface tension. The liquid is able to move up the filter paper because its
attraction to itself is stronger than the force of gravity.

CHROMATOGRAPHY TERMS:-
 ANALYTE:-
The analyte is the substance to be separated during chromatography. It is also normally what is
needed from the mixture.
Analytical chromatography is used to determine the existence and possibly also the
concentration of analyte(s) in a sample.
 BONDED PHASE:-
A stationary phase chemically bonded to a support that is used for the separation.
 CHROMATOGRAM:-
A chromatogram is the visual output of the chromatograph. In the case of an optimal
separation, different peaks or patterns on the chromatogram correspond to different
components of the separated mixture.
 ELUATE:-
Remove (an adsorbed substance) by washing with a solvent, especially in chromatography.
The eluate is the solvent that carries the analyte.
 IMMOBILIZED PHASE:-
An immobilized phase is a stationary phase that is immobilized the support particles, or on the
inner wall of the column tubing.
 SOLUTE:-
A solute is a solid, liquid, or gas which is dissolved to make a solution. A solute is dissolved in a
solvent to make a solution.
 SOLVENT:-
Solvent is a component of a solution that is present in the greatest amount. It is the substance in
which the solute is dissolved.
 DETECTOR:-
The detector refers to the instrument used for qualitative and quantitative detection of analytes
after separation.

ASSIGNMENT NO 1 Chromatography EXACT

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ASSIGNMENT NO 1 Chromatography EXACT

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What are the different types of chromatography discussed?

What are some common terms used in chromatography?

ASSIGNMENT NO 1

2. THIN-LAYER CHROMATOGRAPHY (TLC):-

3. GAS CHROMATOGRAPHY (GC):-

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6. HIGH PRESSURE LIQUID CHROMATOGRAPHY (HPLC):-

MADE BY: MUHAMMAD ALI (ROLL NO 39)

WHAT IS THE RETENTION FACTOR (RF) ?

ANALYTICAL:- determine chemical composition of a sample.

PREPARATIVE:- purify and collect one or more components of a sample.

PIGMENTATION:- separate pigments due to difference in solubility.

MADE BY: MUHAMMAD ALI (ROLL NO 39)

MADE BY: MUHAMMAD ALI (ROLL NO 39)

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