Gas Chromatography

Gas Chromatography
Gas chromatography (GC) is a common type of chromatography used in analytical chemistry
for separating and analysing compounds that can be vaporized without decomposition.
Typical uses of GC include
• Testing the purity of a particular substance
• Separating the different components of a mixture.
Gas Chromatography is a technique applied for
Separation, identification and quantification of components of a mixture of organic compounds
by selective partitioning between the stationary phase and mobile phase inside a column
followed by sequential elution of separated components.
The technique is suitable for separation of compounds having following characteristics :
• High volatility
• Thermal stability
• Low molecular weights

Background of Gas Chromatography

Gas chromatography is widely used for the separation of organic compounds. Benzene and
cyclohexane (having boiling point 80.1 and 80.8◦C) are not able to be separated by distillation
method but readily separated by gas chromatography.
James & Martin introduced Gas Liquid Chromatography

Types of Gas Chromatography

There are two Types of Gas Chromatography
1) Gas–Solid (adsorption) Chromatography
Chromatography in which stationery phase is solid adsorbent.
Separation is based on the relative adsorption of the components of the mixture on stationary
phase
2) Gas–Liquid (partition) Chromatography
Chromatography in which stationery phase is liquid on an inert support
Separation is based on the relative solubilities of the components of the mixture on stationary
phase
In any case, the sample used in gas chromatography must be either a gas or capable of being
converted into gas at the temperature of the column

Muhammad Zain Tariq MCEC-19-04 BS-Chemistry 7th Semester Session 2019-23

Gas Chromatography

GC Separation
Principle
The source of the mobile phase, i.e ., the carrier gas, is usually a high-pressure gas cylinder
equipped with a pressure regulator. The mixture to be separated is loaded in the injector which
vaporizes it, if necessary, and introduces the gaseous mixture into the column where separation
of the mixture into its components occurs. The separated components flow into the detector
where they are continuously monitored. The output from the detector is an electric signal,
which is amplified and recorded by the recorder.

Instrumentation
• Pressure regulator
• Sample injection port
• Gas chromatography column
• Ovens and heaters for maintaining temperatures of the injection port and the column
• Stationary phase
• Detector
• Signal recorder

Factors affecting GC Separation

The most important factor in gas chromatography is the selection of the proper column
(stationary phase for the particular separation to be attempted.
The nature of the liquid or solid phase will determine the exchange equilibrium with the sample
components;
This will depend on the
• Solubility or adsorbability of the analytes
• The polarity of the stationary phase and sample molecules
• The degree of hydrogen bonding
• Specific chemical interactions.

Muhammad Zain Tariq MCEC-19-04 BS-Chemistry 7th Semester Session 2019-23

Gas Chromatography

Process of GC Separation
• In gas chromatography, the sample is converted to the vapour state (if it is not already a
gas) by injection into a heated port, and the eluent is a gas (the carrier gas).
• The stationary phase is generally a non-volatile liquid or a liquid-like phase supported
on or bonded to a capillary wall or inert solid particles such as diatomaceous earth.
• The sample is rapidly injected by means of a hypodermic syringe through a septum or
from a gas sampling valve.
i. Typically, the injected sample first goes into the inlet/inlet liner and
ii. then the carrier gas carries it to the column.
• The sample injection port, column, and detector are heated to temperatures at which the
sample has a vapour pressure of at least 10 torr, usually about 50◦C above the boiling point
of the highest boiling solute
• The injection port and detector are usually kept somewhat warmer than the column
i. To promote rapid vaporization of the injected sample
ii. To prevent sample condensation in the detector.

• For packed columns, liquid samples of 0.1 to 10 μL are injected, while gas samples of 1
to 10 mL are injected. Gases may be injected by means of a gas-tight syringe or through a
special gas inlet chamber of constant volume (gas sampling valve).
• For capillary columns, volumes of only about 1/100 these sizes must be injected because
of the lower capacity (albeit greater resolution) of the columns.

• Sample splitters are included on chromatographs designed for use with capillary columns
that deliver a small fixed fraction of the sample to the column, with the remainder going
waste. They usually also allow splitless injection when packed columns are used
(split/splitless injectors).

• The carrier gas is a chemically inert gas available in pure forms as argon, helium, or
nitrogen. A highly dense gas gives best efficiency since diffusivity is lower, but a low-
density gas gives faster speed. The choice of gas is often dictated by the type of detector.
Separation occurs as the vapour constituents equilibrate between carrier gas and the
stationary phase.

• Gas chromatography always uses flow-through detectors that automatically detect the
analytes as they elute from the column; the majority of GC detectors are destructive.
• The sample emerges from the column at a constant flow rate. A variety of detectors are
used, the specific response is dependent upon the analyte
• Some detectors contain a reference side and a sampling side. The carrier gas is passed
through the reference side before entering the column and emerges from the column
through the sampling.
• The difference in response of the sampling side relative to the reference side is
processed as the analytical signal.
• The signal, representing the chromatographic peaks is acquired and displayed by a data
system as a function of time.

Muhammad Zain Tariq MCEC-19-04 BS-Chemistry 7th Semester Session 2019-23

Gas Chromatography

• By measuring the retention time (the minutes between the time the sample is injected and
the time the chromatographic peak appears) and comparing this time with that of a standard
of the pure substance, it may be possible to identify the peak
• Agreement of retention times of two compounds does not guarantee the compounds are
identical
• The area under the peak is proportional to the concentration, and so the amount of substance
can be quantitatively determined. The peaks are often very sharp (this requires fast
detectors).
• Chromatography data handling systems usually have automatic detection of peaks, readout
of the peak area and/or peak height, as well as the retention time.
• With complex mixtures, it is not a simple task to identify the many peaks.

GC-MS Gas Chromatography–Mass Spectrometry

Instruments are commercially available in which the gaseous effluent is fed into am
spectrometer where they are ionized, sorted on the basis of their mass-to-charger and identified
based on mass/charge ratio mass (as well as and fragmentation pattern). This important
analytical technique is called gas chromatography–mass spectrometry
The mass spectrometer is a sensitive and selective detector, and when a capillary GC column
(very high resolution) is used (capillary GC–MS), this technique is capable of identifying and
quantifying very complex mixtures of trace substances.
For example, hundreds of compounds may be identified in sewage effluents, and traces of
complex drugs in urine or blood or pollutants in water can be determined. Some 4000
compounds have been identified in cigarette smoke.

Injectors
The sample is rapidly injected by means of a hypodermic syringe through a septum or from a
gas sampling valve.
Types of Injectors
1. S/SL (split/splitless) Injector
A sample is introduced into a heated small chamber via a syringe through a septum – the heat
facilitates volatilization of the sample and sample matrix. The carrier gas then either sweeps
the entirety (splitless mode) or a portion (split mode) of the sample into the column.
• In split mode, a part of the sample/carrier gas mixture in the injection chamber is exhausted
through the split vent. Split injection is preferred when working with samples with high
analyte concentrations (>0.1%)
• Splitless injection is best suited for trace analysis with low amounts of analytes (<0.01%).
In splitless mode the split valve opens after a pre-set amount of time to purge heavier
elements that would otherwise contaminate the system. This pre-set (splitless) time should
be optimized, the shorter time (e.g., 0.2 min) ensures less tailing but loss in response, the
longer time (2 min) increases tailing but also signal.

Muhammad Zain Tariq MCEC-19-04 BS-Chemistry 7th Semester Session 2019-23

Gas Chromatography

2. On-column Injector
The sample is here introduced directly into the column in its entirety without heat, or at a
temperature below the boiling point of the solvent. The low temperature condenses the sample
into a narrow zone. The column and inlet can then be heated, releasing the sample into the gas
phase. This ensures the lowest possible temperature for chromatography and keeps samples
from decomposing above their boiling point.
3. PTV Injector
Temperature-programmed sample introduction was first described by Vogt in 1979. Originally
Vogt developed the technique as a method for the introduction of large sample volumes (up to
250 µL) in capillary GC. Vogt introduced the sample into the liner at a controlled injection
rate. The temperature of the liner was chosen slightly below the boiling point of the solvent.
The low-boiling solvent was continuously evaporated and vented through the split line. Based
on this technique, Poy developed the programmed temperature vaporising injector; PTV. By
introducing the sample at a low initial liner temperature many of the disadvantages of the
classic hot injection techniques could be circumvented.
4. Gas source inlet or gas switching valve
Gaseous samples in collection bottles are connected to what is most commonly a six-port
switching valve. The carrier gas flow is not interrupted while a sample can be expanded into a
previously evacuated sample loop. Upon switching, the contents of the sample loop are inserted
into the carrier gas stream.
5. P/T (Purge-and-Trap) system
An inert gas is bubbled through an aqueous sample causing insoluble volatile chemicals to be
purged from the matrix. The volatiles are ‘trapped’ on an absorbent column (known as a trap
or concentrator) at ambient temperature. The trap is then heated and the volatiles are directed
into the carrier gas stream. Samples requiring pre-concentration or purification can be
introduced via such a system, usually hooked up to the S/SL port.

Gas Chromatography Columns

Columns can be in any shape that will fill the heating oven. Column forms include coiled tubes,
U-shaped tubes, and W-shaped tubes, but coils are most commonly used.
The two types of columns used in GC are
• Packed columns
• Capillary columns.
Packed columns came first and were used for many years. Capillary columns are more
commonly used today, but packed columns are still used for applications that do not require
high resolution or when increased capacity is needed.

Muhammad Zain Tariq MCEC-19-04 BS-Chemistry 7th Semester Session 2019-23

Gas Chromatography

1) Packed Column
Typical packed columns are 1 to 10 m long and 0.2 to 0.6 cm in diameter. Well-packed columns
may have 1000 plates/m, and so a representative 3-m column would have 3000 plates.
Short columns can be made of glass or glass/silica-lined stainless steel, but longer columns
may be made of stainless steel or nickel so they can be straightened for filling and packing.
Columns are also made of Teflon. For inertness, glass is still preferred for longer columns.
The resolution for packed columns increases only with the square root of the length of the
column. Long columns require high pressure and longer analysis times and are used only when
necessary (e.g., analytes that are poorly retained require more stationary phase to achieve
adequate retention). Separations are generally attempted by selecting columns in lengths of
multiples of 3, such as 1 or 3 m. If a separation isn’t complete in the shorter column, then the
next longer one is tried.
The column is packed with small particles that may themselves serve as the stationary phase
(adsorption chromatography) or more commonly are coated with a nonvolatile liquid phase of
varying polarity (partition chromatography). Gas–solid chromatography (GSC) is useful for
the separation of small gaseous species such as H2, N2, CO2, CO, O2, NH3, and CH4 and
volatile hydrocarbons, using high surface area inorganic packings such as alumina (Al2O3) or
porous polymers (e.g., Porapak)
The gases are separated by their size due to retention by adsorption on the particles. Gas–solid
chromatography is preferred for aqueous samples.
The solid support for a liquid phase should have a high specific surface area that is chemically
inert but wettable by the liquid phase.
• It must be thermally stable
• And available in uniform sizes.
The most commonly used supports are prepared from diatomaceous earth, a spongy siliceous
material. They are sold under many different trade names.
• Chromosorb P is a pink-coloured diatomaceous earth prepared from crushed firebrick.
• Chromosorb W is diatomaceous earth that has been heated with an alkaline flux to
decrease its acidity; it is lighter in colour.
• Chromosorb G was the first support expressly developed for GC, combining the good
efficiency and handling characteristics of Chromosorb G while having the low
adsorptive properties of Chromosorb W
Column-packing support material is coated by mixing with the correct amount of liquid phase
dissolved in a low-boiling solvent such as acetone or pentane. About a 5 to 10% coating (wt/wt)
will give a thin layer. After coating, the solvent is evaporated by heating and stirring; the last
traces may be removed in a vacuum.

Muhammad Zain Tariq MCEC-19-04 BS-Chemistry 7th Semester Session 2019-23

Gas Chromatography

2) Open Tubular Columns

Open-tubular columns that today provide extremely high resolution and have become the
mainstay for gas-chromatographic analyses.
These columns are made of thin fused silica (SiO2) coated on the outside with a polyimide
polymer for support and protection of the fragile silica capillary, allowing them to be coiled.
The polyimide layer is what imparts a brownish colour to the columns, and it often darkens on
use.
The inner surface of the capillary is chemically treated to minimize interaction of the sample
with the silanol groups (Si–OH) on the tubing surface, by reacting the Si–OH group with a
silane-type reagent (e.g., DMCS).
The capillaries are 0.10 to 0.53 mm internal diameter, with lengths of 15 to 100 m and can have
several hundred thousand plates, even a million. They are sold as coils of about 0.2 m diameter
Capillary columns offer advantages of high resolution with narrow peaks, short analysis time,
and high sensitivity (with detectors designed for capillary GC) but are more easily overloaded
by too much sample. Split injectors by and large alleviate the overloading problem.

There are three types of open-tubular columns.

Wall-coated open-tubular (WCOT) columns have a thin liquid film coated on and supported
by the walls of the capillary. The walls are coated by slowly passing a dilute solution of the
liquid phase through the columns. The solvent is evaporated by passing carrier gas through the
columns. Following coating, the liquid phase is cross-linked to the wall. The resultant
stationary liquid phase is 0.1 to 0.5 μm thick. Wall-coated open-tubular columns typically have
5000 plates/m. So a 50-m column will have 250,000 plates.

Support coated open-tubular (SCOT) columns, in which solid microparticles coated with
the stationary phase (much like in packed columns) are attached to the walls of the capillary.
These have higher surface area and have greater capacity than WCOT columns. The tubing
diameter of these columns is 0.5 to 1.5 mm, larger than WCOT columns.

Porous layer open-tubular (PLOT) columns, have solid-phase particles attached to the
column wall, for adsorption chromatography. Particles of alumina or porous polymers
(molecular sieves) are typically used. These columns, like packed GSC columns, are useful for
separating permanent gases, as well as volatile hydrocarbons.

The resolution efficiency of open-tubular columns is generally in the order: WCOT > SCOT
> PLOT

Muhammad Zain Tariq MCEC-19-04 BS-Chemistry 7th Semester Session 2019-23

Gas Chromatography

Detectors
Detector Applications Sensitivity Linearity Remarks
Range
Thermal General, Fair, 5–100 ng, Good, except Sensitive to
conductivity responds to 10 ppm–100% thermistors at temperature and flow
all substances higher changes; concentration
temperatures sensitive Prone to
burnout at high sample
concentrations
Flame All organic Very good, Excellent, up Requires very stable gas
ionization substances; 10–100 pg, to 106 flow; response for water
some is 10⁴−10⁶ times weaker
oxygenated 10 ppb–99% than for hydrocarbons;
products mass sensitive
respond
poorly. Good
for
hydrocarbons
Flame Sulfur Very good, 10 Excellent
photometric compounds pg S,
(393 nm),
1 pg P
phosphorus
compounds
(526 nm
Electron All Excellent for Poor Very sensitive to
capture substances halogen impurities and
that have containing temperature changes;
affinity to substancs, quantitative
capture 0.05–1 pg, 50 analysis complicated;
electrons; no ppt—1 ppm concentration sensitiv
response for
aliphatic and
naphthenic
hydrocarbons
Vacuum UV Nearly all Excellent down Good up to Very recently
absorption substances to pg levels 10⁴ introduced detector,
but inert expected to have a wide
gases and universal
and nitrogen application area,
provides
some structural
confirmation based
on spectral match
Mass Nearly all Excellent Excellent Can provide structural
spectrometry substances. and molecular
Depends on weight information
ionization
metho

Muhammad Zain Tariq MCEC-19-04 BS-Chemistry 7th Semester Session 2019-23

Gas Chromatography

Headspace Analysis
Headspace analysis avoids the need for solvent extraction for volatile analytes. A convenient
way of sampling volatile samples for GC analysis is the technique of headspace analysis. A
sample in a sealed vial is equilibrated at a fixed temperature, for example, for 10 min, and the
vapor in equilibrium above the sample is sampled and injected into the gas chromatograph. A
typical 20-mL glass vial is capped with a silicone rubber septum lined with
polytetrafluoroethylene (PTFE). A syringe needle can be inserted to withdraw a 1-mL portion.

Thermal Desorption
In thermal desorption, the volatile analyte is desorbed from the sample by heating and
introduced directly into the GC. Thermal desorption (TD) is a technique in which solid or
semisolid samples are heated under a flow of inert gas. Volatile and semi volatile organic
compounds are extracted from the sample matrix into the gas stream and introduced into a gas
chromatograph. Samples are typically weighed into a replaceable PTFE tube liner, which is
inserted into a stainless steel tube for heating.

Purging and Trapping

Purge-and-trap is a form of headspace analysis in which the volatile analyte is trapped on a
sorbent and then thermally desorbed. The purge-and-trap technique is a variation of thermal
desorption analysis in which volatiles are purged from a liquid sample placed in a vessel by
bubbling a gas (e.g., air) through the sample and collecting the volatiles in a sorbent tube
containing a suitable sorbent. The trapped volatiles are then analyzed by thermally desorbing
them from the sorbent.

Muhammad Zain Tariq MCEC-19-04 BS-Chemistry 7th Semester Session 2019-23