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GAS CHROMATOGRAPHY

Manoj Sigdel

GAS CHROMATOGRAPHY
DEFINITION: Gas Chromatography is a technique in which the
vaporized sample components are separated based on
their partition /adsorption between the mobile phase & a
stationary phase liquid / solid, held in a column.
CLASSIFICATION: Based on the nature of stationary phase, the
methodology is divided into
Gas solid Chromatography
Gas liquid Chromatography.

GSC : Mobile phase


gas
Stationary phase solid
In GSC, when a carrier gas containing analytes is
passed through a column containing solid Stationary
phase, the analytes get adsorbed on to the solid
Stationary phase & the separation is due to
differences in their adsorptive behavior.

GLC : Mobile phase


gas
Stationary phase liquid
In GLC, when a carrier gas containing analytes is
passed through a column containing liquid
Stationary phase, the analytes get distributed
themselves between the liquid Stationary phase &
the carrier gas phase according to their partition
coefficients.

In gas chromatography (GC), the sample is


vaporized and injected onto the head of a
chromatographic column. Elution is brought
about by the flow of an inert gaseous mobile
phase.
The mobile phase does not interact with
molecule of the analyte; its only function is to
transport the analyte through the column.

Criteria for compounds to be analyzed by GC :


Volatility- Unless the component is sufficiently volatile in
nature, it cannot be homogeneously mixed with the mobile
phase, hence volatility is an important criteria. (Requires
analyte to be either naturally volatile or can be
converted to a volatile derivative)

Thermostability- One of the requirement of gas


chromatography is that the sample component must be in the
vapour form so solid and liquid samples are heated to high
temperatures to aid volatlization. Hence the samples
components should be thermostable at high temperatures.

Derivatization increases the thermal stability of the


compound.

The analyte should have a measurable vapour pressure at


the temperature employed.

a) DERIVATIZATION: It is a technique, which helps to improve the


analysis, By this technique, we can convert polar,
less volatile molecule into less polar by more
volatile compound. Also this method increases the
thermal stability of the compound.
Derivatization can be brought about by
Esterification
Acylation
Alkylation
Silylation

ESTERIFICATION: It is usual to prepare derivatives of carboxyl gas like


prostaglandins, analgesic, autocoids has carboxyl group,
which is converted to ester that increases volatility
by decreasing hydrogen bonding.

ACYLATION
When the sample under study contains a phenol, primary or
secondary alcohol or amine, derivatization by acylating is frequently
used.
This was performed by use of acetic anhydride and a catalyst like
acetic acid, p-toluene, sulfonic acid, pyridine, N-methylimidazole

Perfluoroacylation:
Although acetylation is usually adequate to give good gas
chromatographic characteristics, trifluoroacetates,
pentafluoropropionates or heptafluorobutyrates are currently used
to increase the sensitivity of the analysis.This group significantly
increases the mol wt of the sample relative to the analogous
hydrocarbon which is best method to increase the retension time

Alkylation:

Alkylation is used to derivatize functional groups such as


alcohols, phenols, amines, imides, and sulfhydryl groups
in which there is labile hydrogen.
Alkylation can be accomplished by using the Williamson
ether synthesis in which an alcohol or phenol is
treated with an alkyl or benzyl halide in the presence
of base

SILYLATION: The active hydrogen of OH, - COOH, - NH2, SH, can be replaced by silyl groups to give more
volatile derivatives.
Several silylating agents are used & all of them
have a general formula (CH3)3 SiX
R OH + (CH3)3 SiCl
(Trimethyl chlorosileane)

R O Si (CH3)3 + HCL
(Tri methyl silyl ether)

Uses of Derivatization:

Increase volatility (i.e,sugars)

Eliminates the presence of polar groups like OH,NH and SH.

Derivatization targets O, S, N, & P functional


groups.

Increases detectability i.e., steroids/ cholesterol.

THEORY OF GAS CHROMATOGRHPHY:


The two theories have been established to describe the
gas chromatographic separation. They are: a) Plate theory
b) Rate theory
a) Plate theory: - According this theory, the
chromatographic column consists of a
number of continuos-narrow, horizontal plates called the
Theoritical Plates which are the hypothetical units of the
column.

In plate theory it is assumed that, during


chromatographic process: All the solute particles are placed in the first
theoretical plate &
Equillibration of the solute particles between the 2
phases takes place at each theoretical plate. This theory
measures efficiency of the column & the factors which
influence the efficiency are
1) The number of theoretical plates (N)
2) Height equivalent of theoretical plates (H)
3) Resolution (R)

b) RATE THEORY
Plate theory failed to explain the ways to improve the
performance of the column, which the rate theory did.
This theory explained the fact that the phase flows
continuously & that the solute particles are constantly
being transported & partitioned in the column.
It can be explained by Vandeemeter Equation
H = A+B + C i) A Eddys diffusion
ii) B Longitudinal diffusion
iii) C Mass transfer resistance

Instrumentation
Whether GLC or GSC consists of basic
components are,
1) A carrier gas, which is maintained at a high
pressure and is delivered to the instrument
at a rapid and reproducible rate.
2) Flow meters.
3) Sample injection system.
4) Separation column.
5) One or more detectors.
6) Thermostated chambers for the temperature
regulation of the column & detector.

Carrier gas
The choice of the carrier gas is some time
dependent upon the type of the detector used in gas
chromatography. Most widely used carrier gases are
nitrogen, helium, hydrogen, and argon.
Hydrogen
Better thermal conductivity,
Low density.
It is not useful often as the other gases because it is
associated with fire hazards and it react with
unsaturated compounds.
Helium
Excellent thermal conductivity, but it expensive.
Is the one of the gas most widely employed.
Nitrogen

Ideal characteristics of the carrier gas;

Inertness
Suitable to the detector used
High purity
Easily available
Cheap
Less risk of the explosion
Should give best column performance
consistent with the required speed of the
analysis.

Flow meters
As we know that the carrier gas are
stored at high pressure. In order to
maintain the uniform flow rate of carrier
gas we use flow meters &flow regulator.
Flow meters are used to measure the
flow rate of carrier gas. They are
Rotameter & Soap bubble meter.

Rotameter is placed conveniently


before the column inlet. It has an
ordinary glass tube like burette with a
float held on to a spring. The level of
the float is determined by the flow rate
of the carrier gas & is pre calibrated.

Soap bubble meter

It is also similar to Rotameter and instead of a float, a soap


bubble is formed which indicates the flow rate. It has glass
tube with a inlet at the bottom through which gas comes in.
A rubber bulb is used to store the soap solution. When the
bulb is gently pressed, a drop of soap solution is formed
which is converted into bubble by the pressure of carrier
gas & travels up. The distance traveled upward is a
measure of the flow rate of the carrier gas. The graduations
are precalibrated.

Sample injection system :


Samples for introducing into the column can be of any type i.e, either
gas ,liquid or solid in nature.
*Gases - by syringe or by loop injector.

*Liquids can be
injected through loop /septum devices. Usually a rubber septum
made up of silicone rubber which can withstand high temp is used.

*Solid samples are dissolved in solvent and then injected into the
septum.
The ideal introduction of sample into the gas ' chromatography system
would be to inject it as a compact "plug" onto the first theoretical
plate of the column.

Column
Two categories of column are used for gas
chromatography.
a) Packed column
b) Open tubular column or capillary column

a) Packed column are those which contain


particles of the stationary phase packed into a
metallic or glass tube. The metallic tubes are
usually constructed of stainless steel, but copper
and aluminium tubes are also used.
Diameter: 2 to 20 mm
Length: 1 to 4 meters
The length of the packed column is dependent
upon the required number of theoretical plates.
To fit into the temperature controlled oven in the
gas chromatograph the packed column is
usually carefully bent or coiled. The bending or
coiling is done such a way that it should not

The stationary phase in the packed column is


either small particles of a solid adsorbent for
GSC or a liquid supported on small particles of
a solid in GLC.
The function of the solid support in GLC packed
column is to provide an inert surface on which
the liquid phase can be coated and to provide
an appropriate physical structure for the
stationary phase.
The solid support must be thermostable at the
temperature at which column is operated and it

Solid support is derived from diatomaceous


earths. They are the skeletal remains of the
unicellular algae known as diatoms. They
consist of hydrated silica groups. The large
surface area of treated diatomaceous earths is
nearly ideal as solid supports for the GLC.
The stationary phase used in GLC packed
column and in some open tubular column must
be a non-volatile liquid at the temperature at
which the column is operated. The sample
compounds must be at least slightly soluble in
the liquid phase in order to be separated, and
the liquid should, if possible, be a pure

Polar substances are usually better separated


on a polar stationary phase and non polar
substances on non polar stationary phases.
Stationary phases
Nature
Temperature in use
Poly dimethyl siloxane Nonpolar
-60 to 3200C
Poly(diphenyl)dimeth Nonpolar bonded phase -60 to 3200C
yl siloxane
Polycyano propyl
phenyl dimethyl
siloxane
Polyalkylene glycol
Polyethylene glycol

intermediate polarity

upto 2800C

Polar
Polar

30 to2200C
50 to 2800C

Packed Columns

The major advantage and use


is for large-scale or
preparative purification

Industrial scale purification


maybe
in the kilogram or
Oil refinery

separatesgreater range

fractions of
oil for
petroleum
products

500 L
chromatograp
hy column

b) Open tubular columns constitutes the second category of


GC columns. Rather than being filled with particles as packing
material, they have the liquid stationary phase coated on the
columns inner wall. These column are also called as capillary
column or Golay columns
Because the pressure drop through open tubular column is
considerably less then that through packed columns, it is
possible to use open tubular columns, which are longer, then
packed columns.
Length: 10 to 100 meter
Diameter: 0.2 to 0.5 mm.
Material of construction: glass or fused silica

Metals are avoided because they catalyze the several


chemical reactions.
Operating Temperature: 10 to 25 oC above the boiling point of
the highest boiling sample component.
There are two main types :
Wall coated open tubular (WCOT), in which the
stationary phase is coated directly on to the inner walls
of tubing, which is liquid.
Support coated open tubular (SCOT),which have a finely
divided layer of solid support material deposited on the inner
walls onto which the stationary phase is then coated. Though
not as efficient as WCOT, they have a higher sample capacity,
which enable them to be used without a stream splitter.

Advantages of open columns over packed column


Diameter of the column is small and the number of different
paths through the column that sample component can follow is
minimized
The coating on WCOT and bonded phase columns
generally are more uniform then the stationary phase used
in packed column.
All of those factors combine to produce better resolution on
coated capillary columns then on packed columns.

Column temperature control:


Control of column temperature is important in order to
produce reproducible chromatogram. It is done by GC Oven
which can be set to operate at fix temp.
Accurate control of column temperature is necessary in
isothermal analysis. To avoid problems of the least volatile
component in a sample having component of different
volatile in a mixture, which takes long time to elute, the
column temperature can be progressively increased while
the flow rate is kept constant.

Detectors
Characteristics of a good detector
o
o
o
o
o
o
o

Sensitivity appropriate to sample


Large linear dynamic range
Useful at a range of temperatures
Rapid response time
Easy to use
Stable, Predictable response
Nondestructive (probably least important)

In general detectors can be classified into two groups


Differential detectors
Integrating detectors

Differential Detectors
Those detectors that respond to the conc. (in
mole fraction) of solute in the carrier gas. Eg:
Thermal conductivity & electron capture
detectors, Photo Ionization Detectors

Advantages:
Do not destroy the sample in the process of
detecting it.
Thus collection of successive fractions of the
solute for the further characterization is
possible.
Disadvantages:
Precise or efficient quantitative analysis is

Integrating Detectors
Those detectors that respond to the mass flow rate
of the
solute in moles per unit time. Eg:
flame ionization detectors, flame photometric
detectors.

Advantages:
Precise or efficient quantitative analysis is
possible.
Disadvantages:
destroy the sample in the process of detecting
it.

Comparison Of Detectors
DETECTOR

TYPE SUPPORT SELECTIVITY


GAS

DETECT DYNAM
IBILITY IC
RANGE

FID (Flame
Ionization
Detector )

Mass
flow

100pg

107

TCD (Thermal
Conductivity
Detector )

Conc. Reference Universal

1ng

107

ECD
(Electron
Capture
Detector )

Con.

PID (Photo
Ionization
Detectors )

Conc, Make-up

Aliphatic,aromatic, 2pg
ketones,esters,ald
ehydes,amines

FPD (Flame
Photometric
Detectors )

Mass
flow

SO4,PO4,tin,boron,
arsenic,germaniu
m,selenium,chrom
ium

Hydrogen
& air

Make-up

Hydrogen
& air or
oxygen

Most organic
compounds

Halides,nitrates,nit 50pg
riles,peroxides,anh
ydriedes,organom
etallics

100pg

105

10

103

Interfacing Gas Chromatography


with Spectroscopic Methods
Gas chromatography is often coupled with
the selective techniques of spectroscopy,
thus giving so-called hyphenated methods
that provide the chemist with powerful tools
for identifying the components of complex
mixtures.
Gas Chromatography/Mass
Spectrometry (GC/MS)
The flow rate from capillary columns is
generally low enough that the column
output can be fed directly into the ionization
chamber of the mass spectrometer. For
packed columns and megabore capillary

Thank You