Chromatographic methods

Gas chromatography

Shishir Kanti Pramanik, PhD

Professor
Department of Chemistry, SUST

Dept. of Chemistry, SUST

 INTRODUCTION

• Gas chromatography (GC) was discovered by Mikhail Semenovich Tsvett

(early 1900s)

• Used to analyze volatile substances in the gas phase

• The components of a sample are dissolved in a solvent and vaporized

in order to separate the analytes by distributing the sample between two
phases: a stationary phase and a mobile phase.

• Process of separating component(s) from the given sample mixture by

using a gaseous mobile phase. The mobile phase is a chemically inert
gas that serves to carry the molecules of the analyte through the heated
column

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 INTRODUCTION

• It involves a sample being vaporized and injected onto the head of

the chromatographic column. The sample is transported through the
column by the flow of inert, gaseous mobile phase. The column
itself contains a liquid stationary phase or solid stationary phase
which is adsorbed onto the surface of an inert Solid

• The stationary phase is either a solid adsorbant, termed gas-solid

chromatography (GSC), or a liquid on an inert support, termed
gas-liquid chromatography (GLC)

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 Types of GC
Two major types:

• Gas-solid chromatography (GSC):

✓ Using a packed column with a solid surface of particles forming
stationary phase, e.g., alumina or cross-linked polymer
✓ Mobile phase is a gas
✓ Used for separation of low molecular gases, e.g., air components, H2S,
CS2 ,CO2, rare gases, CO and oxides of nitrogen

• Gas-liquid chromatography (GLC):

✓ Using a packed column with a liquid stationary phase coated onto inert
support particles, and stationary phase retains onto by adsorption or
chemical bonding
✓ The mobile phase is a gas
✓ Using open tubular column with the liquid stationary or solid stationary
phase coated onto inner walls of the column tubing referred to as wall
coated (WCOT), porous layer open tubular column (PLOT) and surface
coated open tubular column (SCOT)
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 Principles

• The principle of separation in GC is “partition.”

• The analytes to be separated is distributed accordion to That mean K=
Csp/Cmp.
• Distribution ratio depend on the analytes vapor pressure,
thermodynamics properties of the bulk component band and affinity for
the stationary phase.
Mobile Phase Vapor pressure

Component Distribution according to K and

temperature
Stationary Phase Affinity for SP

• The mixture of component to be separated is converted to vapour and

mixed with gaseous mobile phase.
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 Principles

• The component which is more soluble in stationary phase travel slower

and eluted later. The component which is less soluble in stationary
phase travels faster and eluted out first.
• No two components has same partition coefficient conditions. So the
components are separated according to their partition coefficient.
• Partition coefficient is “the ratio of solubility of a substance distributed
between two immiscible liquids at a constant temperature.

To
detector

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 Instrumentation

1. Carrier Gas
2. Flow regulators and flow Meters
3. Injection Devices
4. Columns
5. Temperature Control Devices
6. Detectors
7. Recorders And Integrators

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GC main components

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 Carrier gas

Hydrogen-Good thermal conductivity, low density, inflamable

Helium - Good thermal conductivity, but expensive
Nitrogen - Inexpensive, Reduced sensitivity

The carrier gas acts as the mobile phase and transport the sample
component through the column to the detector, retardation occurring due to
interaction with the stationary phase

It should meet the following criteria:

• Should be chemically inert to the column materials and sample
components
• Gases with the smallest diffusion coefficients, hydrogen, helium will give
better separation efficiencies than higher molecular weight gases such as
N2
• Ratio of viscosity to diffusion coefficient should be minimum for rapid
analysis
• Carrier gas purities should be better than 99.99%
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 Criteria for Carrier gas

• Should be cheap and readily available

• Should be of high quality and not cause any fire accidents
• Should give best possible results
• Should be suitable for the sample to be analyzed and for the detector
• Hydrogen has low density and better thermal conductivity. However, it
reacts with unsaturated compounds and is inflammable and explosive in
nature.
• Nitrogen is inexpensive but it gives reduced sensitivity.
• He is the most preferred gas.
• Inlet pressure ranges from: 10-50 psi
• -Flow rate : 25-150 mL/min for packed columns
• Flow rate: 2-25 mL/min for open tubular column
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 INJECTION DEVICES (Sampling unit)

• Sampling unit or injection port is attached to the column head.

• Since the sample should be in vapourized state, the injection port is
provided with an oven that helps to maintain its temperature at about
20-500 C above the boiling point of the sample.
• Gaseous samples may be introduced by use of a gas tight hypodermic
needle of 0.5-10 ml capacity.
• For Liquid samples , micro syringes of 0.1-100μL capacity may be
used.
Direct injection is mostly used.
Types:
1.Automatic sampler
2.Headspace Sampling
3.Purge and Trap
Automatic sampler
4.Pyrolysis
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 COLUMN UNIT

• Most important part of the GC

• Columns are of different shapes and sizes that includes: “U” tube type
or coiled helix type
• They are mainly made of stainless steel, and Glass

➢ Support material
✓ it’s main function is to provide mechanical support to the liquid
phase. An ideal support should have a large surface area,
chemically inert, should get uniformly wet with liquid phase, should
be thermostable.
✓ Commonly used solid phases are: diatomaceous earth or
kieselguhr, glass beads, porous polymers, sand,etc.

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➢ Liquid phase:-
It should have the following requirements:
• It should be non-volatile
• Should have high decomposition temperature
• Should be chemically inert
• Should posses low vapour pressure at column temperature
• Should be chemically and structurally similar to that of the solute
i.e., polar for polar solute.

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 COLUMN UNIT

Examples of different liquid phases:

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 COLUMN UNIT

Types of columns:-
• There are two general types of columns:

1. Packed columns:- In GLC, they are densely packed with finely

divided, inert, solid support material(diatomaceous earth) coated with
liquid stationary phase.
-In GSC, the columns are packed with adsorbents or porous polymers.
✓ Length: 1.5 - 10m
✓ internal diameter: 2 - 4mm.

2. Capillary columns-
✓ length ranges from 10-100m
✓ inner diameter is usually 0.1-0.5mm.

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 COLUMN UNIT

Capillary columns-
It is mainly of two types:
❑Wall-coated columns - consist of a capillary tube whose walls are
coated with liquid stationary phase.

❑ Support-coated columns- the inner wall of the capillary is lined

with a thin layer of support material such as diatomaceous earth, onto
which the stationary phase has been adsorbed. It is also known as
PLOT (porous-layer open tubular columns).
➢SCOT columns are generally less efficient than WCOT columns. Both
types of capillary column are more efficient than packed columns.

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COLUMN UNIT

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 COLUMN UNIT

Equilibrium of the column:-

• The packed columns are equilibrated before introduction of the sample.
This is done by allowing continuous flow of heated carrier gas through the
columns for a specific duration of time (24hrs) at prescribed temperature.
• Ideally prepared and conditioned columns show a zero base line on the
recorder upon passage of carrier gas alone.

➢Column temperature:-
• This can be controlled by jackets equipped with vapours of a boiling liquid,
electrically heated metal blocks or circulating air baths.
• Compounds of low B.P- eluted at lower temperature
• Compounds of high B.P- boils at higher temperature resulting in broader
and shallower peaks, require temperature programming.
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➢ Column oven
serves to control the temperature of the column within a few tenths
of a degree to conduct precise work

▪ Can be operated in two manners:

✓ isothermal programming or
✓ temperature programming

▪ Isothermal programming
• Temperature of the column is held constant throughout the entire separation.
• The optimum column temperature for isothermal operation is about the
middle point of the boiling range of the sample.
• Isothermal programming works best only if the boiling point range of the
sample is narrow.
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▪ Isothermal programming
▪ Problems:
1. If a low isothermal column temperature is used with a wide boiling
point range, the low boiling fractions are well resolved but the high
boiling fractions are slow to elute with extensive band broadening
2. If the temperature is increased closer to the boiling points of the
higher boiling components, the higher boiling components elute as
sharp peaks but the lower boiling components elute so quickly there
is no separation.

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 Liquid phase Chromatography

➢ Temperature programming method

✓ Column temperature is either increased continuously or in steps as the
separation progresses.
✓ The method is well suited to separating a mixture with a broad boiling
point range.
✓ The analysis begins at a low temperature to resolve the low boiling
components and increases during the separation to resolve the less
volatile, high boiling components of the sample.
✓ Rates of 5-7 °C/minute are typical for temperature programming
separations.

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 Sampling Techniques for GC
Necessity of the Derivatisation of sample
Some compounds are not sufficiently volatile, so they tail badly and are too strongly
attracted to the stationary phases .
• To increase the volatility of the sample
• To reduce thermal degradation of the sample by increasing thermal stability
• To increase the detector response by incorporating into the derivative
functional groups which produce a higher detector signal
• To improve separation and reduce tailing
Derivatisation methods
1. Silylation
• Replacement of acidic hydrogen on the analyte molecule with an alkylsilyl group
(SiMe3)
• Derivatives are generally less polar, more volatile and more thermally stable

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 Sampling Techniques

Derivatisation methods

2. Acylation

• Used to form perfluroacyl derivatives of alcohols and phenols or amines

• To enhance detector performance using an electron capture detector as well as
increase volatility

i)

ii)

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 Sampling Techniques

Derivatisation methods

3. Alkylation (esterification)

• Addition of the alkyl group to an active functional group, formation of methyl esters
• Useful derivatisation reaction

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