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….
