Gas Chromatography

Outline

• Chromatography
• Classification of Chromatography
• Gas Chromatography (GC)
• Mass Spectrometry
• Gas Chromatography-Mass Spectrometry (GC–
MS)
 Chromatography
• Is a technique used to separate and identify
the components of a mixture.

• Works by allowing the molecules present in

the mixture to distribute themselves between
a stationary and a mobile medium.

• Molecules that spend most of their time in the

mobile phase are carried along faster.
 Chromatography

• Chromatography, in its various forms, is a purification and

analytical technique that has the widest applicability in
organic chemistry

• Virtually all stable molecules will survive one or more types

of chromatographic separation

• Depending upon the system used, the purity of

chromatographed compounds can be from modest to very
high – even to analytical purity
 Tswett’s Original
History Experiment

• Chromatography was discovered by Michael Tswett

in the first part of the 20th century - in 1906. Powdered
Sucrose
Column
• The technique was discovered in the course of
investigating plant pigments

• Tswett passed pentane solutions of plant pigments

through a bed of powdered sucrose and observed
the formation of colored bands – hence the name
chromatography.

• At the time he speculated that it would be possible

to separate colorless compounds by the same
 Chromatography

• Chromatography is a physical method of

separation

• Components to be separated are

distributed between two phases

• One phase is stationary while the other

phase is mobile which moves through it
in a definite direction

• The chromatographic process occurs due

to differences in the distribution
constant of the individual sample
 Chromatography
• The separation of the different components in
the mixture is based on
• Polarity
• Boiling point
• Ionic strength
• Size
 Chromatography
• Mobile phase: phase in which sample is dissolved in;
may be gas, liquid or supercritical fluid

• Stationary phase: phase through which mobile

phase is forced to pass

• Mobile and stationary phases are chosen and the

analyte will distribute itself between the two phases
 Classification of Chromatography

According to the force of separation

1. Adsorption chromatography
2. Partition chromatography
3. Ion exchange chromatography
4. Gel filtration chromatography
5. Affinity chromatography
 According to the packing of the stationary phase

1. Thin layer chromatography (TLC):

 The stationary phase is a thin layer supported on
glass, plastic or aluminium plates

2. Paper chromatography (PC):

 The stationary phase is a thin film of liquid
supported on an inert support

3. Column chromatography (CC):

 The stationary phase is packed in a glass column
 According to the mobile phase

• Liquid chromatography:
mobile phase is a liquid (LLC, LSC)

• Gas chromatography:
mobile phase is a gas (GSC, GLC)
 Different Types of chromatography
Mechanism Mobile phase Stationary phase Mode or type
Solutes move at different rates Liquid or gas Solid that attracts Adsorption
according to the forces of attraction the solutes Chromatography
.to the stationary phase
Solutes equilibrate between the 2 Liquid or gas Thin film of liquid Partition
phases according to their partition formed on the Chromatography
coefficients surface of a solid
inert support
Solute ions of charge opposite to the Liquid Solid resin that Ion Exchange
fixed ions are attracted to the resin containing carries fixed ions Chromatography
by electrostatic forces & replace the electrolytes & mobile
.mobile counterions couterions of
opposite charge
attached by
covalent bonds

Molecules separate according to Liquid Porous gel with no Molecular Exclusion

:their size attractive action Chromatography
1.Smaller molecules enter the pores on solute
of the gel, and need a larger volume molecules
of eluent.
2.Larger molecules pass through the
column at a faster rate.
Special kind of solute molecules interact with Liquid or gas Solid on which specific Affinity
those immobilized on the stationary phase molecules are
immobilized Chromatography
 What is GC ?
• GC is a Separation Technique
• Sample is usually a complex mixture we require
to separate into constituent components

• Usually to quantify some or all components e.g.

Pharmaceuticals, Environmental pollutants, etc.

• Occasionally used as a qualitative tool

 Gas - Solid Chromatography (GSC)
• The stationary phase, in this case, is a solid like silica or
alumina

• It is the affinity of solutes towards adsorption onto the

stationary phase which determines, in part, the
retention time

• The mobile phase is, of course, a suitable carrier gas

• The use of GSC in practice is considered marginal when
compared to gas liquid chromatography
 Gas - Liquid Chromatography (GLC)

• Partition of molecules between gas (mobile phase) and

liquid (stationary phase).
• The stationary liquid phase with very low volatility is held
on particles of a solid support.
• The mobile phase is a suitable carrier gas (Helium,
Nitrogen)
• GLC is the most widely used technique for the separation
of volatile species
• The wide variety of stationary phases are available
• Column preparation is quite easy
• GLC is simply called GC
In the animation below the red molecules are more soluble in the liquid
(or less volatile) than are the green molecules.
 Schematic Diagram of Gas Chromatography
Injection
port Recorder

Oven

Detector

Column
Nitrogen
cylinder
 Gas Chromatography
Filters/Traps Data system
H

RESET

Regulators Syringe/Sampler

Inlets

Detectors • gas system

Gas Carrier
Hydrogen

• inlet
Air

Column • column
• detector
• data
system
 Chromatogram of petrol

Some additional peaks (unlabeled) are due to the

presence of other fragments present in the petrol
 Most Common Stationary Phases

 Separation of mixture of polar

compounds Carbowax 20M
(polyethylene glycol)

 Separation of mixtures of non-polar

compounds OV101 or SE-30 (polymer of
methylsilicone)

 Methylester of fatty acids DEGS

(diethylene glycol succinate)
 GC Components
• Carrier Gas, N 2 or He, 1-2 mL/min
• Injector
• Oven
• Column
• Detector
 Syringe

Injector
Detector

Gas tank

Column

Oven
 Carrier Gas

Inert

Helium

Choice dictated by detector, cost, availability

Pressure regulated for constant inlet pressure

Flow controlled for constant flow rate

Chromatographic grade gases (high purity)

 Injector
• A GC syringe penetrates a septum to inject sample into
the vaporization chamber

• Instant vaporization of the sample, 280 C

• Carrier gas transports the sample into the head of the
column

• Purge valve controls the fraction of sample that enters

the column
 Oven

• Programmable
• Isothermal- run at one constant temperature
• Temperature programming - Start at low
temperature and gradually ramp to higher
temperature
• More constant peak width
• Better sensitivity for components that are retained
longer
• Much better chromatographic resolution
• Peak refocusing at head of column
 Column
It is normally contained in a Thermostatic Oven
About 1μL of liquid is injected into one end of the column

As each component reaches the other end it is detected and registered

on a chart recorder

The Retention Time (the time required to travel through the column) is
characteristic of a particular substance (for the same column, temperature,
gas flow etc.)

The area under each peak indicates the relative quantities

 Open Tubular Capillary Column

0.32 mm ID
Mobile phase
(Helium)
flowing at 1 Liquid
mL/min Stationary 0.1-5 mm
phase

15-60 m in length
Fused Silica Open Tubular (FSOT) Columns

• Coated with polymer, crosslinked

• Polydimethyl siloxane (non-polar)
• Poly(phenylmethyldimethyl) siloxane (10% phenyl)
• Poly(phenylmethyl) siloxane (50% phenyl)
• Polyethylene glycol (polar)
• Poly(dicyanoallyldimethyl) siloxane
• Ploy(trifluoropropyldimethyl) siloxane
 Polar vs Non-polar
• Separation is based on the vapor pressure and
polarity of the components.

• Within a homologous series (alkanes, alcohols,

olefins, fatty acids) retention time increases with
chain length (or molecular weight)

• Polar columns retain polar compounds to a

greater extent than non-polar
• C18 saturated vs. C18 saturated methyl ester
 Detectors

• Thermal Conductivity Detectors (TCD)

• Flame Ionization Detectors (FID)
• Electron Capture Detectors (ECD)
• Electron impact/chemical ionization (EI/CI)
 Advantages and Disadvantages of GC

Advantages Disadvantages
• Fast analysis • Limited to volatile samples
- Typically minutes (even sec.) T limited to ~ 380 °C
• High Resolution Need Pvap ~ 60 Torr at
- Record N~1.3 x 106 that temperature
• Sensitive detectors (easy ppm, often
ppb)
• Highly accurate quantification (1-5 %
• Not suitable for thermally
labile samples
RSD)
• Automated systems • Some samples may require
extensive preparation
• Non-destructive*
- Allows online coupling to Mass Spec. • Requires spectroscopy
(usually MS) to confirm peak
• Small sample (mL)
identify
• Reliable and relatively simple
• Low cost
 Thermal Conductivity Basics
The TCD is a nondestructive, When the carrier gas is contaminated
concentration sensing detector. A by sample , the cooling effect of
heated filament is cooled by the the gas changes. The difference in
flow of carrier gas cooling is used to generate the
detector signal
Flow

Flow
 Thermal Conductivity Detector
Principle: The thermal balance of a heated filament

• When a compound elutes, the thermal conductivity

of the gaseous mixture of carrier gas and compound
gas is lowered, and the filament in the sample
column becomes hotter than the other control
column.

• Its resistance increased, and this imbalance between

control and sample filament resistances is measured
by a simple gadget and a signal is recorded
 Flame Ionization Detectors (FID)
• Effluent exits column and enters an air/hydrogen
flame

• The gas-phase solute is pyrolized to form electrons

and ions

• All carbon species are reduced to CH 2

+
ions

• These ions collected at an electrode held above the

flame

• The current reaching the electrode is amplified to

give the signal
 Characteristics of FID

• A general detector for organic compounds

• Very sensitive (10-13 g/s)
• Linear response (10 7
)
• Rugged
• Disadvantage: specificity
 Electron Capture Detector (ECD)

• Ultra-sensitive detection of
halogen-containing species

• For pesticide analysis (picogram)

• Accept electrons of carrier gas

 Electron Capture Detector

• ECD detects ions in the exiting from the gas chromatographic

column by the anode electrode.
• 3
H or 63Ni which emits  particles.
• Ionization : N2 (Nitrogen carrier gas) +  (e) = N2+ + 2e

• These N2+ establish a “base line”

• X (F, Cl and Br) containing sample +  (e)  X-
• on recombination : X- + N2+ = X + N2
• The “base line” will decrease and this decrease constitutes
the signal.
• Insecticides, pesticides, vinyl chloride and fluorocarbons are
very easily detected
Electron Capture Detector
 Function
• Separation of volatile organic compounds
• Volatile – when heated, VOCs undergo a phase
transition into intact gas-phase species

• Separation occurs as a result of unique equilibria

established between the solutes and the stationary
phase (the GC column)

• An inert carrier gas carries the solutes through the

column