PRINCIPLE, METHODOLOGY AND

APPLICATION OF GAS
CHROMATOGRAPHY (GC)
CHAPTER 7
BMS481
Lesson Learning Outcome (LLO)

At the end of this course, students should be able to

 1. Understand the principle of GC
 2. Know the application and limitation of GC
 3. Compare between GC and LC
Outline

 Uses of Gas Chromatography

 GC components and Types of Columns
 Factors Affecting Chromatographic Separation
 Data Analysis
 Which Industries
Use
Chromatography?

 Chemical/Petrochemical
 Clinical/Forensic
 Consumer Products
 Environmental
 Food
 Pharmaceutical
Why Gas Chromatography?

 Simple
 Cheap (can be automated)
 Short analysis times
 High accuracy
 Qualitative and Quantitative analysis
 Applicable in % to ppb level
 Should be thermally stable

Compounds
Should be un-reactive and
Amenable to non-absorptive to
GC???? chromatographic system

Should be volatile Presence of

polar groups
at temperature reduces
below 350-400 C volatility
IUPAC Definition of Chromatography

“ A physical method of separating sample components from a mixture by

selective adsorption or partitioning of the analyte between two phases:

A “mobile phase and a stationary phase”

Types of GC and their phases
• 1. GLC – Gas Liquid Chromatography
• Stationary phase: immobilized liquids
(siloxanes, polyethylene glycols..)
• Principle: Partition

• 2. GSC – Gas Solid Chromatography

• Stationary phase : Solids (alumina, silica,
polymers, carbon…)
• Principle : Adsorption

*Mobile phases
Gases (N2, He, H2, Ar)…GC
GC Principle


The principle of GC involves separation ofvolatile components of the
sample based on their distribution and partition co-efficient between two
phases.
 This is ratio of solubility of substance in between gaseous mobile phase and
stationary liquid phase.
 The components of the sample that are partitioned into gas come out first
while others come later
How gas chromatography works

 GC consists of a carrier gas flow, an injector port, a

column, a temperature regulated oven and a
detector.

 The sample is injected into the heated injection port

(to be volatilised) and is carried by the mobile
gas into the column. The column is heated to provide
sufficient vapor pressure to elute the sample. Inside
the column, the components get separated by the
differential partition in between the mobile phase
gas and stationary phase liquid. The separated
components flow through a heated detector for
observation.
GC components and types of columns

 Components of a Gas Chromatography

 Types of GC columns
 Types of GC capillary columns
Components of a Gas Chromatograph
Mobile phases (carrier gas)

N2, He and H2 are typical carrier gases

 He:
- Most common and compatible with most detectors
- Better resolution (smaller plate heights)
- Solutes diffuse rapidly  smaller mass transfer term
 N2:
- Lower detection limit for a flame ionization detector
- Lower resolution and solute diffusion rates
 H2:
- Inexpensive
- Fastest separations (optimum velocity is 2x higher
than He)
Flow rate increases N2 < He < H2
- Can not be used with mass spectrometers might
explosive mixtures with air
- Better resolution (smaller plate heights)
- Solutes diffuse rapidly  smaller mass transfer term
Injection port
• The sample to be analyzed is loaded at the injection
port via a hypodermic syringe.
• The injection port is heated (350oC )in order to volatilize
the sample
• Once in the gas phase, the sample is carried to mixing
chamber for complete vaporization mixing before
entering column
• GC is very sensitive instrument. Typically samples of
one micro liter or less are injected on the column
• These volumes can be further reduced by using what is
called a split injection system in which a controlled
fraction of the injected sample is carried away by a gas
stream before entering the column.
Types of GC columns
Open tubular (Capillary) columns

 Commonly used in GC
 Higher resolution, shorter analysis time, and greater sensitivity
 Low sample capacity
Capillary column
materials
 Metal Tubing- MXT
 Stainless steel
 Almost as inert as fused silica but useable up
to approx. 450 °C

 Fused Silica
 Synthethic, amorphous glass
 Excellent inertness and useable to approx.
380 °C (400 °C)
Types of Capillary
columns
 WCOT
 Partition chromatography
 Typical phases: Siloxanes and Polyethylene glycols
 0.10 to 0.53mm internal diameters
 PLOT
 Adsorption chromatography
 Gas, light hydrocarbons/solvent analysis
 Adsorbents: porous polymers, alumina (<1 um particle
diameter)
 0.25 to 0.53mm internal diameter
 The critical parameters for GC column:
Column
• Dimensions : internal diameter, column length, film
selection thickness
• Conditions: temperature, flow rate
parameters • Composition: stationary phase composition, carrier gas

Given a sample, you will need first to choose

what stationary phase will work best
• First pick the type of column, then think about dimension
• Conditions can be optimized for given column dimension

Choice of stationary phase is very important

• It determines what kind of sample you can run

• Critical for packed columns, but less so for open tube
column because of its high efficiency
Factors impacting the separation
Non-column factors affecting separation

 Carrier gas: type & linear velocity

 Temperature
 Injection bandwidth
 Column factors: diameter, length
 Column temperature

 Increase in temperature gradient during

separation, increase the solute vapor
pressure and hence decrease the retention
time
 Stationary phase factor:

Basic rule in GC stationary phase…

“choose a stationary phase that “looks like” the

components you want to separate..”
The detector
Qualitative and Quantitative Analysis
 Mass Spectrometer and Fourier Transform Infrared Spectrometers can identify compounds as part of a GC
system
- Compare spectrum with library of spectra using a computer

 Compare retention times between reference sample and unknown

- Use multiple columns with different stationary phases
- Co-elute the known and unknown and measure changes in peak area

 The area of a peak is proportional to the quantity of that compound

Peak area increases proportional

Peak Area

to concentration of standard if
unknown/standard have the
identical retention time  same
compound

Concentration of Standard
Type of detectors

 Thermal conductivity detector

 Two columns with conducting wire in between
 No sample – the thermal conduction is constant (temperature of
both gases is the same)

 Injected sample - The effluent gas carries the

sample components into the detector column. Since effluent
gas is mixed with sample components there results in the
difference in thermal conductivity from prior one recording.

This difference in conductivity is specific for the component

analyzed. This is recorded for further comparison and
identification of the components and their quantity.
 Flame ionisation detector

 Sample components from effluent are

ionized by subjecting to flame in a
chamber
 These ions raise upwards and are attracted
towards anode or catode based on the
charge on them
 When the impinge on the electrodes, the
current is passed which is recorded.
 The strength and intensity of current
depending on the sample and is specific
 Useful for organic compunds, atmospheric
and aqueous environmental samples
 Sample is destroyed
 GC Analysis
*the elution

 Order of elution
(retention time) is
mainly determined by
volatility
 The volatility also
depends on polarity of
the compound and
between the stationary
phase
Polar compound = least volatile = most retained
Polar compound = polar stationary phase = most retained
 The number of components in
a sample is determined by the
number of peaks
 The amount of a given
component in a sample is
determined by the area under
the peaks
 The identity of components
can be determined by the
given retention times – Must
be couple with another
instrument (e.g. MS)
THANK YOU

TEST 2
IS WAITING….