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CHROMATOGRAPHY - Class #5

The document discusses separation methods, including solvent extraction and chromatography. It covers the types of solvent extraction and chromatography, including thin layer chromatography, gas chromatography, and high performance liquid chromatography. It also discusses principles, terminology, factors that affect efficiency, and applications of these separation methods. The document provides examples of numerical problems related to solvent extraction and chromatography calculations.

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Brijesh Sharma
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72 views

CHROMATOGRAPHY - Class #5

The document discusses separation methods, including solvent extraction and chromatography. It covers the types of solvent extraction and chromatography, including thin layer chromatography, gas chromatography, and high performance liquid chromatography. It also discusses principles, terminology, factors that affect efficiency, and applications of these separation methods. The document provides examples of numerical problems related to solvent extraction and chromatography calculations.

Uploaded by

Brijesh Sharma
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
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RAMAIAH

College of Arts,
Science &
Commerce

SEPARATION METHODS
Class #5
(21-05-2021)
Solvent Extraction – Types, continuous, efficiency, selectivity. Distribution coefficient,
Nernst distribution law, derivation factors affecting the partition, applications.
Chromatography – Types, Terminology, Principles of Paper, Thin layer, Column, gas
chromatography, column efficiency, plate theory, factors affecting the column efficiency,
band broadening. Rf factor, Van-Demeter equation, medium performance, liquid
chromatography, high performance liquid chromatography. Reverse phase liquid
chromatography, super critical fluid chromatography, characteristics of supercritical fluids.
2D-thin layer chromatography, electrophoresis, principles, applications etc., Numerical
problems on solvent extraction, Rf factor and van-Demeter equation.

Dr. Vasanth K
RAMAIAH
College of Arts,
Science &
Commerce
SUPERCRITICAL FLUIDS
SUPERCRITICAL FLUID (SCF)

 The behavior of SCF was first observed in 1822 by CHARLES CAGNIARD

 SCF can be any substance at a TEMPERATURE and PRESSURE above its


CRITICAL POINT where distinct liquid and gas phase do not exist.

 Hence, at the critical point or stationary point is a point on PHASE


DIAGRAM at which both LIQUID and GAS phases of a substance have the
same density and are hence indistinguishable.

 Thus when a compound is subjected to PRESSURE and TEMPERATURE


higher than its CRITICAL POINT, the fluid is said to be SUPRCRITICAL.

 At this point or region, fluid exhibit with INTERMEDIATE properties between


liquid and gas. They possess liquid-like DENSITIES and gas-like
VISCOSITIES.

 Thus, the fluid is said to be SUPERCRITICAL when it is heated above its


CRITICAL TEMPERATURE and compressed above its CRITICAL PRESSURE.
Dr. Vasanth K
RAMAIAH
College of Arts,
Science &
Commerce

SUPERCRITICAL FLUIDS

Dr. Vasanth K
RAMAIAH
College of Arts,
Science &
Commerce

SUPERCRITICAL FLUIDS

Dr. Vasanth K
RAMAIAH
College of Arts,
Science &
Commerce

SUPERCRITICAL FLUIDS

Dr. Vasanth K
RAMAIAH
College of Arts,
Science &
Commerce

SUPERCRITICAL FLUIDS
 EXAMPLES:

1. SUPERCRITICAL CO2:

VAPOUR

LIQUID+VAPOUR

Dr. Vasanth K
RAMAIAH
College of Arts,
Science &
Commerce
SUPERCRITICAL FLUIDS
 Carbon dioxide is most widely used SCF, as it is Cheaper, Chemically INERT,
Non-toxic, non-flammable and is readily available.

 Critical temperature of CO2 : 31oC


Critical pressure of CO2: 74 bar (7.4 pascals)

USES:

 Supercritical CO2 is known as GREEN SOLVENT and being non-toxic it is


used for extraction purposes in various food industries and pharmaceutical
extractions.

 It is used to remove organochlorides pesticides and metal impurities from


agriculture crops and herbal industries.

Dr. Vasanth K
RAMAIAH
College of Arts,
Science &
Commerce
SUPERCRITICAL FLUIDS

2. SUPERCRITICAL WATER:

Critical temperature: 150 to 250oC


Critical pressure: 15 to 100 bar

 SC water can solubilize hydrophobic compounds hence it is also known as


HOT COMPRESSED WATER. This SCF water is used for EXTRACTION of
plant materials, waste processing (wet air oxidation), etc.

 In SCF water, organic compounds and gases become highly miscible and
precipitation of inorganic compounds occur. It is also used to synthesize
nanoparticles.

3. Other supercritical fluids like ETHANE, PROPANE, METHANOL, ETHANOL


are also used in its supercritical phase for various extractions, synthesis and in
chemical reactions.
Dr. Vasanth K
RAMAIAH
College of Arts,
Science &
Commerce

SUPERCRITICAL FLUID CHROMATOGRAPHY (SFC)

PRINCIPLE:

 This technique is also known as CONVERGENCE CHROMATOGRAPHY.

 SFC is a form of NORMAL PHASE chromatography that uses


SUPERCRITICAL FLUID such as CO2 as the mobile phase.

 SFC is used to analyze and purify the LOW to HIGH molecular weight
molecules, thermally labile molecules and also used to separate the
CHIRAL COMPOUNDS.

 SFC is particularly useful for analysis of non-volatile and easily


thermally-decomposable samples. This is not suitable for samples used
for spectroscopy, GC or HPLC.

Dr. Vasanth K
RAMAIAH
College of Arts,
Science &
Commerce

SUPERCRITICAL FLUID CHROMATOGRAPHY (SFC)

INSTRUMENT:

Dr. Vasanth K
RAMAIAH
College of Arts,
Science &
Commerce
SUPERCRITICAL FLUID CHROMATOGRAPHY (SFC)
Mobile Phase:
 Majority of applications in SFC and in supercritical extraction (SFE) uses
CO2 as the principal component of mobile phase.

 This is because the critical conditions of CO2 (Pc: 7.4 Mpa; and Tc: 31.3oC)
are mild, and CO2 is cheaper and non-toxic.

 To increase the solubility of solute in mobile phase, the CO2 is often mixed
with modifiers like (Eg: Methanol, ethanol, isopropanol, n-propanol,
acetonitrile, n-hexane,, THF, etc) with or without additives like organic
base or acid.

 This can be used for both normal and as a reverse phase.

Conditions:
 Temperature and Pressure are constant at approximately 25oC and 15 Mpa
(Supercritical Conditions).

 In SFC, CO2 pumps are kept under refrigeration as gas is pumped


Dr.in liquid K
Vasanth
RAMAIAH
College of Arts,
Science &
Commerce

SUPERCRITICAL FLUID CHROMATOGRAPHY (SFC)


Stationary Phase and Columns:

 Columns of 25 X 0.46 cm packed with 5 µm particles whose efficiency is in


between gas chromatography and HPLC.

 CO2 is non-polar and hence it can solubilize hydrophobic compounds


when this mobile phase is used. Hence, for hydrophobic solutes bonded
alkyl stationary phases are used, thereby acting as a reverse phase liquid
chromatography (RPLC).

 Similarly polar solutes can be separated by using polar stationary phase


(Normal phase chromatography) Eg: silica, aminopropyl, poly ethtylene
glycol – bonded phases etc

 Staionary phases like (R) – N – (3,5 – dinitrobenzol) phenyl glycine


covalently bonded to amino propyl silica with CO2 and modifiers like
methanol, ethanol or 2-propanol conditions are used for separation of
CHIRAL COMPOUNDS.
Dr. Vasanth K
RAMAIAH
College of Arts,
Science &
Commerce

SUPERCRITICAL FLUID CHROMATOGRAPHY (SFC)


DETECTORS:

1. UV//Visible detectors
2. Flame ionization detectors (FID)
3. Evaporative light scattering detectors

APPLICATIONS:

1. SFC is used as an interphase technique combined with mass spec used


for analysis of the pharmaceutical products, natural products, etc.
2. It is mainly used to characterize and purify CHIRAL/ACHIRAL compound
3. It can be used as both reversible and normal phase chromatography to
separate non-polar an polar solutes respectively
4. It has application in pharmaceutical, agrochemical, propellant, explosive,
forensic, food processing, metallurgical etc type of industries.

Dr. Vasanth K
RAMAIAH
College of Arts,
Science &
Commerce

Dr. Vasanth K

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