Theory and Instrumentation of GC

Introduction to Gas Chromatography

Learning Aims & Objectives

Aims

Outline a Brief History of Gas Chromatography (GC)
Compare and contrast GC with other analytical techniques primarily High
Performance Liquid Chromatography (HPLC)
Explain the function of each major component of the GC system
Explain the terms and appearance of a typical Chromatogram
Outline the fundamental basis for separation in GC
Indicate the major advantages of GC and the application areas in which it is used


Objectives
At the end of this Section you should be able to:

Identify analytes suitable for GC analysis from physico-chemical data
Describe the function of the various components of a Gas Chromatograph
Explain the fundamental basis of separation in GC in terms of solubility and vapour
pressure of analytes
Recognise when the use of GC might be applicable to solving analytical problems








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Content


Origins of Gas Chromatography 3
Why Choose Gas Chromatography? 4
Why Choose Gas Chromatography? 6
GC Separation Mechanism 7
The Distribution Constant (Partition Coefficient) (Kc) 7
The Gas Chromatograph 8
The GC instrument process 8
The Chromatogram 9
GC Advantages & Disadvantages 10
Typical GC Applications 10




3
Origins of Gas Chromatography
Gas chromatography was invented by A. J. P. Martin who, with R. L. M. Synge, suggested
its possibility in a paper on liquid chromatography published in 1941.



When dealing with liquid-liquid partition chromatography, they predicted that the mobile
phase need not be a liquid but may be a vapour. Very refined separations of volatile
substances in a column in which a permanent gas is made to flow over gel impregnated
with a non-volatile solvent would be much faster and thus, the columns much more
efficient and separation times much shorter.
So the concept of gas chromatography was envisioned more than fifty years ago, but
unfortunately, little notice was taken of the suggestion and it was left to Martin himself and
his co-worker A. T. James to bring the concept to practical reality some years later in
1951, when they published their epic paper describing the first gas chromatograph.
They demonstrated the technique by separating and quantitatively determining the
components of a C1-C12 fatty acid mixture. The importance of GC was recognized almost
immediately by petrochemical laboratories, which faced the challenge of analysing
complex hydrocarbon mixtures.
















An early GC separation of a fatty acid mixture by Martyn and James


Archer J.P. Martin Richard L.M. Synge
(1910-2002) (1914-1994)

4
Why Choose Gas Chromatography?

The two main chromatographic techniques used in modern analytical chemistry are Gas
Chromatography (GC) and High Performance Liquid Chromatography (HPLC).

A typical HPLC chromatograph (left) and a Gas Chromatograph (right)

HPLC uses a liquid mobile phase to transport the sample components (analytes) through
the column, which is packed with a solid stationary phase material.


Typical HPLC column (left) and GC column (right)

HPLC was first proposed by the Russian botanist Mikhail Tswett first used the term
Chromatography (Latin for coloured drawing) in 1906, to describe the separation that
occurred when solutions of plant pigments were passed through columns of calcium
carbonate or alumina, using petroleum ether.

In contrast, Gas Chromatography uses a gaseous mobile phase to transport sample
components through either packed columns or hollow capillary columns containing the
stationary phase. In most cases, GC columns have smaller internal diameter and are
longer than HPLC columns.

GC has developed into a sophisticated technique since the pioneering work of Martin and
James in 1952, and is capable of separating very complex mixtures of volatile analytes.


5


Schematic diagram of a typical capillary Gas Chromatograph

Gas Inlets:
Gas is fed from cylinders through supply piping to the instrument. It is usual to filter gases
to ensure high gas purity and the gas supply pressure. Required gases might include:
Carrier (H
2
, He, N
2
)
Make-up gas (H
2
, He, N
2
)
Detector fuel gas (H
2
& Air/Ar or Ar and CH
3
/N
2
)
depending on detector type.

Injector:
Here the sample is volatilised and the resulting gas entrained into the carrier stream
entering the GC column.
Many inlet types exist including:
Split/Splitless,
Programmed Thermal Vaporising (PTV),
Cool-on-column (COC) etc.
The COC injector introduces the sample into the column as a liquid to avoid thermal
decomposition or improve quantitative accuracy.

Detector:
The detector responds to a physico-chemical property of the analyte, amplifies this
response and generates an electronic signal for the data system to produce a
chromatogram. Many different types exist and the choice is based mainly on application,
analyte chemistry and required sensitivity also on whether quantitative or qualitative data
is required.

Detector choices include: Flame ionisation (FID) / Electron Capture (ECD) / Flame
Photometric (FPD) / Nitrogen Phosphorous (NPD) / Thermal Conductivity (TCD) and
Mass Spectrometer (MS)

Data System:
The data system receives the analogue signal from the detector and digitises it to form the
record of the chromatographic separation known as the Chromatogram.

The data system can also be used to perform various quantitative and qualitative
operations on the chromatogram assisting with sample identification and quantitation.


6
Pneumatic controls:
The gas supply is regulated to the correct pressure (or flow) and then fed to the required
part of the instrument. Control us usually required to regulate the gas coming into the
instrument and then to supply the various parts of the instrument. A GC fitted with a Split /
Splitless inlet, capillary GC column and Flame Ionisation detector may have the following
different gas specifications:

Carrier gas supply pressure / Column Inlet Pressure (column carrier gas flow) / Inlet split
flow / Detector make-up gas flow. Modern GC instruments have Electronic Pneumatic
pressure controllers older instruments may have manual pressure control via regulators.


Why Choose Gas Chromatography?

The following information gives an indication of the type of sample (analyte) analysed by
either GC and HPLC and relative strengths and limitations of each technique.

GC
Samples analysed by GC must be volatile (have a significant vapour pressure
below 250
o
C)
Derivatisation to increase volatility is possible but can be cumbersome and
introduces possible quantitative errors
Most GC analytes are under 500 Da Molecular Weight for volatility purposes
Highly polar analytes may be less volatile than suspected when dissolved in a
polar solvent or in the presence of other polar species due to intermolecular forces
such as hydrogen bonding.

HPLC
HPLC analysis has no volatility issues, however the analyte must be soluble in the
mobile phase.
HPLC can analyse samples over a wide polarity range and is able to analyse
ionic samples. Mobile phase components are selected to ensure sample solubility.
HPLC has no real upper molecular weight limit and large proteins of many
thousands of Daltons may be analysed.
So under what circumstances would we chose GC to separate our sample
components?

Table 1. Molecular properties of selected analytes
Molecule Name Properties GC suitability

Hexane C
6
H
14
(86.2Da)
Boiling point: 69
o
C
Vapour pressure (@25
o
C)
130kPa
Yes

Benzene C
6
H
6
(78.1Da)
Boiling point: 80.1
o
C
Vapour pressure (@25
o
C)
12.7kPa
Yes

Anthracene C
6
H
14
(178.1Da)
Boiling point: 340
o
C
Vapour pressure (@25
o
C) n/a.
kPa
No



7
GC Separation Mechanism

In Gas Chromatography (GC) the mobile phase is a gas and the stationary phase is either
a solid - Gas solid chromatography (GSC) or an immobilised polymeric liquid - Gas Liquid
Chromatography (GLC). Of the two types of GC, GLC is by far the most common as will
be seen.

The Distribution Constant (Partition Coefficient) (Kc)

[ ]
[ ] Cm
Cs
Kc =

Where
Cs refers to the concentration of analyte in the stationary phase.
Cm refers to the concentration of analyte in the mobile phase.

The Distribution Coefficient measures the tendency of an analyte to be attracted to the
stationary phase. Larger Kc values lead to longer retention analyte times. The value of
Kc can be controlled by the chemical nature of the stationary phase and the column
temperature.

The figure shows a typical separation process in GC. Each sample component partitions
between the gaseous mobile phase and liquid stationary phase (often coated onto the
inner wall of a long thin capillary tube). The rate and degree of partitioning depends upon
the chemical affinity of the analyte for the stationary phase and the analyte vapour
pressure which is governed by the column temperature.












The diagram shows that analyte A has a higher affinity for the mobile phase (lower Kc
value) and therefore elutes more quickly than analyte B.

From the figure it can be seen that component A has a lower affinity for the stationary
phase and therefore is moved through the column more quickly than
component B, which spends more of its time in the stationary phase in this way
separation is achieved.

In GC, analyte separation is achieved by optimising the differences in stationary phase
affinity and the relative vapour pressures of the analytes. In practice these parameters are
manipulated by changing the chemical nature of the stationary phase and the column
temperature.




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8
The Gas Chromatograph

Instrumentation for Gas Chromatography has continually evolved since the inception of
the technique in 1951 and the introduction of the first commercial systems in 1954.

Most modern commercial GC systems operate in the following way:

An inert carrier gas, such as helium, is supplied from gas cylinders to the GC
where the pressure is regulated using manual or electronic (pneumatic) pressure
controls.
The regulated carrier gas is supplied to the inlet and subsequently flows through
the column and into the detector.
The sample is injected into the (usually) heated injection port where it is volatilised
and carried into the column by the carrier gas.
The sample is separated inside the column - usually a long silica based column
with small internal diameter. The sample separates by differential partition of the
analytes between the mobile and stationary phases, based on relative vapour
pressure and solubility in the immobilised liquid stationary phase.
On elution from the column, the carrier gas and analytes pass into a detector,
which responds to some physico-chemical property of the analyte and generates
an electronic signal measuring the amount of analyte present.
The data system then produces an integrated chromatogram.

The GC instrument process

The sample is injected into the inlet where it is volatilised and a representative
portion is carried onto the column by the carrier gas.
The sample components are separated by differential portioning in the stationary
and mobile phases.
The separated sample components elute from the column into the detector where
some physico-chemical parameter is detected and a signal produced.
This signal is then amplified and sent to the data system where the chromatogram
is electronically constructed.





















9
The Chromatogram

As the components elute from the column they pass into a detector where some
physico-chemical property of the analyte produces a response from the detector. This
response is amplified and plotted against time giving rise to a chromatogram.































Typical chromatogram


Components (such as the injection solvent) that are not retained within the column elute
at the dead time or hold up time t
M
. There are various ways of measuring this parameter
using unretained compounds such as methane or hexane.

Those compounds (analytes and sample components) that are retained elute as
approximately Gaussian shaped peaks later in the chromatogram. Retention times
provide the qualitative aspect of the chromatogram and the retention time of a compound
always will be the same under identical chromatographic conditions. The chromatographic
peak height or peak area is related to the quantity of analyte. For determination of the
actual amount of the compound, the area or height is compared against standards of
known concentration.





10
GC Advantages & Disadvantages

Gas chromatography has several important advantages which are listed opposite.

GC techniques produce fast analyses because of the highly efficient nature of the
separations achieved this will be studied later on. It can be argued that modern GC
produces the fastest separations of all chromatographic techniques. A column has been
produced to separate 970 components within a reasonable analysis time - proving that
very complex separations may be carried out using GC.

By using a combination of oven temperature and stationary phase chemistry (polarity)
very difficult separations may also be carried out including separations of chiral and
other positional isomers.

GC is excellent for quantitative analysis with a range of sensitive and linear detectors to
choose from.

GC is however limited to the analysis of volatile samples. Some highly polar analytes can
be derivatised to impart a degree of volatility, but this process can be difficult and may
incur quantitative errors.

A practical upper temperature limit for conventional GC columns is around 350-380
o
C.
Analyte boiling points rarely exceed 400
o
C in GC analysis and the upper Molecular Weight
is usually around 500 Da.

Advantages
Fast analysis.
High efficiency leading high resolution.
Sensitive detectors (ppb).
Non destructive enabling coupling to Mass Spectrometers (MS) an instrument
that measures the masses of individual molecules that have been converted
into ions, i.e., molecules that have been electrically charged.
High quantitative accuracy (<1% RSD typical)
Rugged and reliable thechiques.
Well established with extensive literature and applications.
Disadvantages
Limited to volatile samples.
Not suitable for samples that degrade at elevated temperatures (thermally labile).
Not suited to preparative chromatography.
Requires MS detector for analyte structural elucidation (characterisation).
Most non-MS detectors are destructive.


Typical GC Applications

Since the development of GC instruments in the early to mid 1950s, GC has found
applications in a host of industrial, environmental, pharmaceutical and biotechnology
analytical laboratories.

Modern GC techniques are able to sample from a wide variety of matrices, including
solids, liquids and permanent gases.


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High temperature applications using specially designed columns are able to analyse
relatively non-volatile substances and Cool-on-Column injection techniques allow the
sampling of moderately thermally labile materials.
Purge and trap and headspace autosampling techniques are now well established and are
able to desorb or extract samples collected in the most inhospitable of environments, such
as the emission stacks of industrial plants.

Detector technology for GC is able to detect very small amounts of pesticides for example,
from environmental samples and GC-MS techniques allow structural elucidation of even
the most complex analytes.

Click on each of the application areas opposite to get more specific details on applications
by industrial sector.

Pharmaceutical
In the pharmaceutical industry GC is used to analyse residual solvents in
both raw materials (drug substance) and finished products (drug product).
Biopharmaceutical applications include urine drugs screens for
barbiturates and underivatised drugs ethylene oxide in sterilized products
as sutures.


Foods/Flavours/Fragrances
The food industry uses GC for a wide variety of applications including
quality testing and solvents testing. The Flavours and Fragrances
industries use GC for quality testing and fingerprinting of fragrances for
characterization.



Petrochemical
GC applications include natural gas analysis or refineries, gasoline
characterization and fraction quantitation, aromatics in benzene, etc.
Geochemical applications include mapping of oil reserves and tracing of
reservoir etc.


Environmental
Environmental GC applications include detection of pollutants such as
pesticides, fungicides, herbicides, purgeable aromatics, etc.
Industrial environmental protection applications include stack and waste
emissions as well as water discharges.



Chemical/Industrial
Chemical/Industrial uses include determination of product content,
determination of purity, monitoring production process, etc GCs are used
to detect organic acids, alcohols, amines, esters, and solvents.