Gas Chromatography

Gas chromatography is a term used to describe the group of analytical separation

techniques used to analyze volatile substances in the gas phase. In gas chromatography, the
components of a sample are dissolved in a solvent and vaporized in order to separate the
analytes by distributing the sample between two phases: a stationary phase and a mobile
phase. The mobile phase is a chemically inert gas that serves to carry the molecules of the
analyte through the heated column. Gas chromatography is one of the sole forms of
chromatography that does not utilize the mobile phase for interacting with the analyte. The
stationary phase is either a solid adsorbant, termed gas-solid chromatography (GSC), or a
liquid on an inert support, termed gas-liquid chromatography (GLC).

Introduction
In early 1900s, Gas chromatography (GC) was discovered by Mikhail Semenovich Tsvett
as a separation technique to separate compounds. In organic chemistry, liquid-solid column
chromatography is often used to separate organic compounds in solution. Among the
various types of gas chromatography, gas-liquid chromatography is the method most
commonly used to separate organic compounds. The combination of gas chromatography
and mass spectrometry is an invaluable tool in the identification of molecules. A typical gas
chromatograph consists of an injection port, a column, carrier gas flow control equipment,
ovens and heaters for maintaining temperatures of the injection port and the column, an
integrator chart recorder and a detector.

To separate the compounds in gas-liquid chromatography, a solution sample that contains

organic compounds of interest is injected into the sample port where it will be vaporized.
The vaporized samples that are injected are then carried by an inert gas, which is often used
by helium or nitrogen. This inert gas goes through a glass column packed with silica that is
coated with a liquid. Materials that are less soluble in the liquid will increase the result
faster than the material with greater solubility. The purpose of this module is to provide a
better understanding on its separation and measurement techniques and its application.

In GLC, the liquid stationary phase is adsorbed onto a solid inert packing or immobilized
on the capillary tubing walls. The column is considered packed if the glass or metal column
tubing is packed with small spherical inert supports. The liquid phase adsorbs onto the
surface of these beads in a thin layer. In a capillary column, the tubing walls are coated
with the stationary phase or an adsorbant layer, which is capable of supporting the liquid
phase. However, the method of GSC has limited application in the laboratory and is rarely
used due to severe peak tailing and the semi-permanent retention of polar compounds
within the column. Therefore, the method of gas-liquid chromatography is simply
shortened to gas chromatography and will be referred to as such here. The purpose of this
module is to provide a better understanding on its separation and measurement techniques
and its application.
Instrumentation
Sample Injection

A sample port is necessary for introducing the sample at the head of the column. Modern
injection techniques often employ the use of heated sample ports through which the sample
can be injected and vaporized in a near simultaneous fashion. A calibrated microsyringe is
used to deliver a sample volume in the range of a few microliters through a rubber septum
and into the vaporization chamber. Most separations require only a small fraction of the
initial sample volume and a sample splitter is used to direct excess sample to waste.
Commercial gas chromatographs often allow for both split and splitless injections when
alternating between packed columns and capillary columns. The vaporization chamber is
typically heated 50 °C above the lowest boiling point of the sample and subsequently
mixed with the carrier gas to transport the sample into the column.

Figure 1: A cross-sectional view of a microflash vaporizer direct injector.

Carrier Gas

The carrier gas plays an important role, and varies in the GC used. Carrier gas must be dry,
free of oxygen and chemically inert mobile-phase employed in gas chromatography.
Helium is most commonly used because it is safer than, but comprable to hydrogen in
efficiency, has a larger range of flow rates and is compatible with many detectors.
Nitrogen, argon, and hydrogen are also used depending upon the desired performance and
the detector being used. Both hydrogen and helium, which are commonly used on most
traditional detectors such as Flame Ionization(FID), thermal conductivity (TCD) and
Electron capture (ECD), provide a shorter analysis time and lower elution temperatures of
the sample due to higher flow rates and low molecular weight. For instance, hydrogen or
helium as the carrier gas gives the highest sensitivity with TCD because the difference in
thermal conductivity between the organic vapor and hydrogen/helium is greater than other
carrier gas. Other detectors such as mass spectroscopy, uses nitrogen or argon which has a
much better advantage than hydrogen or helium due to their higher molecular weights, in
which improve vacuum pump efficiency.

All carrier gasses are available in pressurized tanks and pressure regulators, gages and flow
meters are used to meticulously control the flow rate of the gas. Most gas supplies used
should fall between 99.995% - 99.9995% purity range and contain a low levels (< 0.5 ppm)
of oxygen and total hydrocarbons in the tank. The carrier gas system contains a molecular
sieve to remove water and other impurities. Traps are another option to keep the system
pure and optimum sensitive and removal traces of water and other contaminants. A two
stage pressure regulation is required to use to minimize the pressure surges and to monitor
the flow rate of the gas. To monitor the flow rate of the gas a flow or pressure regulator was
also require onto both tank and chromatograph gas inlet. This applies different gas type will
use different type of regulator.The carrier gas is preheated and filtered with a molecular
sieve to remove impurities and water prior to being introduced to the vaporization chamber.
A carrier gas is typically required in GC system to flow through the injector and push the
gaseous components of the sample onto the GC column, which leads to the detector ( see
more detail in detector section).

 
Figure 3. Gas Recommendations for Packed Columns

Column Oven
The thermostatted oven serves to control the temperature of the column within a few tenths
of a degree to conduct precise work. The oven can be operated in two manners: isothermal
programming or temperature programming. In isothermal programming, the temperature of
the column is held constant throughout the entire separation. The optimum column
temperature for isothermal operation is about the middle point of the boiling range of the
sample. However, isothermal programming works best only if the boiling point range of the
sample is narrow. If a low isothermal column temperature is used with a wide boiling point
range, the low boiling fractions are well resolved but the high boiling fractions are slow to
elute with extensive band broadening. If the temperature is increased closer to the boiling
points of the higher boiling components, the higher boiling components elute as sharp
peaks but the lower boiling components elute so quickly there is no separation.

In the temperature programming method, the column temperature is either increased

continuously or in steps as the separation progresses. This method is well suited to
separating a mixture with a broad boiling point range. The analysis begins at a low
temperature to resolve the low boiling components and increases during the separation to
resolve the less volatile, high boiling components of the sample. Rates of 5-7 °C/minute are
typical for temperature programming separations.

Figure 3. The effect of column temperature on the shape of the peaks.

Open Tubular Columns and Packed Columns

Open tubular columns, which are also known as capillary columns, come in two basic
forms. The first is a wall-coated open tubular (WCOT) column and the second type is a
support-coated open tubular (SCOT) column. WCOT columns are capillary tubes that have
a thin later of the stationary phase coated along the column walls. In SCOT columns, the
column walls are first coated with a thin layer (about 30 micrometers thick) of adsorbant
solid, such as diatomaceous earth, a material which consists of single-celled, sea-plant
skeletons. The adsorbant solid is then treated with the liquid stationary phase. While SCOT
columns are capable of holding a greater volume of stationary phase than a WCOT column
due to its greater sample capacity, WCOT columns still have greater column efficiencies.

Most modern WCOT columns are made of glass, but T316 stainless steel, aluminum,
copper and plastics have also been used. Each material has its own relative merits
depending upon the application. Glass WCOT columns have the distinct advantage of
chemical etching, which is usually achieved by gaseous or concentrated hydrochloric acid
treatment. The etching process gives the glass a rough surface and allows the bonded
stationary phase to adhere more tightly to the column surface.

One of the most popular types of capillary columns is a special WCOT column called the
fused-silica wall-coated (FSWC) open tubular column. The walls of the fused-silica
columns are drawn from purified silica containing minimal metal oxides. These columns
are much thinner than glass columns, with diameters as small as 0.1 mm and lengths as
long as 100 m. To protect the column, a polyimide coating is applied to the outside of the
tubing and bent into coils to fit inside the thermostatted oven of the gas chromatography
unit. The FSWC columns are commercially available and currently replacing older columns
due to increased chemical inertness, greater column efficiency and smaller sampling size
requirements. It is possible to achieve up to 400,000 theoretical plates with a 100 m WCOT
column, yet the world record for the largest number of theoretical plates is over 2 million
plates for 1.3 km section of column.

Packed columns are made of a glass or a metal tubing which is densely packed with a solid
support like diatomaceous earth. Due to the difficulty of packing the tubing uniformly,
these types of columns have a larger diameter than open tubular columns and have a limited
range of length. As a result, packed columns can only achieve about 50% of the efficiency
of a comparable WCOT column. Furthermore, the diatomaceous earth packing is
deactivated over time due to the semi-permanent adsorption of impurities within the
column. In contrast, FSWC open tubular columns are manufactured to be virtually free of
these adsorption problems.

Figure 4. Properties of gas chromatography columns.

 
 Figure 5. Computer Generated Image of a FSWC column (specialized for measuring BAC
levels)

Figure 6. Computer Generated Image of a FSWC column (specialized to withstand extreme

heat)

Different types of columns can be applied for different fields. Depending on the type of
sample, some GC columns are better than the others. For example, the FSWC column
shown in Figure 5 is designed specially for blood alcohol analysis. It produces fast run
times with baseline resolution of key components in under 3 minutes. Moreover, it displays
enhanced resolutions of ethanol and acetone peaks, which helps with determining the BAC
levels. This particular column is known as Zebron-BAC and it made with polyimide
coating on the outside and the inner layer is made of fused silica and the inner diameter
ranges from .18 mm to .25 mm. There are also many other Zebron brand columns designed
for other purposes.

Another example of a Zebron GC column is known as the Zebron-inferno. Its outer layer is
coated with a special type of polyimide that is designed to withstand high temperatures. As
shown in figure 6, it contains an extra layer inside. It can withstand up to 430 °C to be exact
and it is designed to provide true boiling point separation of hydrocarbons distillation
methods. Moreover, it is also used for acidic and basic samples.
Detection Systems
The detector is the device located at the end of the column which provides a quantitative
measurement of the components of the mixture as they elute in combination with the carrier
gas. In theory, any property of the gaseous mixture that is different from the carrier gas can
be used as a detection method. These detection properties fall into two categories: bulk
properties and specific properties. Bulk properties, which are also known as general
properties, are properties that both the carrier gas and analyte possess but to different
degrees. Specific properties, such as detectors that measure nitrogen-phosphorous content,
have limited applications but compensate for this by their increased sensitivity.

Each detector has two main parts that when used together they serve as transducers to
convert the detected property changes into an electrical signal that is recorded as a
chromatogram. The first part of the detector is the sensor which is placed as close the the
column exit as possible in order to optimize detection. The second is the electronic
equipment used to digitize the analog signal so that a computer may analyze the acquired
chromatogram. The sooner the analog signal is converted into a digital signal, the greater
the signal-to-noise ratio becomes, as analog signal are easily susceptible to many types of
interferences.

An ideal GC detector is distinguished by several characteristics. The first requirement is

adequate sensitivity to provide a high resolution signal for all components in the mixture.
This is clearly an idealized statement as such a sample would approach zero volume and the
detector would need infinite sensitivity to detect it. In modern instruments, the sensitivities
of the detectors are in the range of 10 -8 to 10-15 g of solute per second. Furthermore, the
quantity of sample must be reproducible and many columns will distort peaks if enough
sample is not injected. An ideal column will also be chemically inert and and should not
alter the sample in any way. Optimized columns will be able to withstand temperatures in
the range of -200 °C to at least 400 °C. In addition, such a column would have a short linear
response time that is independent of flow rate and extends for several orders of magnitude.
Moreover, the detector should be reliable, predictable and easy to operate.

Understandably, it is not possible for a detector meet all of these requirements. The next
subsections will discuss some of the more common types of gas chromatography detectors
and the relative advantages and/or disadvantages of each.

Table 7: Typical gas chromatography detectors and their detection limits.

Type of Detector Applicable Samples Detection Limit

Mass Spectrometer (MS) Tunable for any sample .25 to 100 pg

Flame Ionization (FID) Hydrocarbons 1 pg/s

Table 7: Typical gas chromatography detectors and their detection limits.

Type of Detector Applicable Samples Detection Limit

Thermal Conductivity (TCD) Universal 500 pg/ml

Electron-Capture (ECD) Halogenated hydrocarbons 5 fg/s

Atomic Emission (AED) Element-selective 1 pg

Dark current of
Chemiluminescence (CS) Oxidizing reagent
PMT
.002 to .02
Photoionization (PID) Vapor and gaseous Compounds
µg/L
 

Mass Spectrometry Detectors

Mass Spectrometer (MS) detectors are most powerful of all gas chromatography detectors.
In a GC/MS system, the mass spectrometer scans the masses continuously throughout the
separation. When the sample exits the chromatography column, it is passed through a
transfer line into the inlet of the mass spectrometer . The sample is then ionized and
fragmented, typically by an electron-impact ion source. During this process, the sample is
bombarded by energetic electrons which ionize the molecule by causing them to lose an
electron due to electrostatic repulsion. Further bombardment causes the ions to fragment.
The ions are then passed into a mass analyzer where the ions are sorted according to their
m/z value, or mass-to-charge ratio. Most ions are only singly charged.

The Chromatogram will point out the retention times and the mass spectrometer will use
the peaks to determine what kind of molecules are exist in the mixture. The figure below
represents a typical mass spectrum of water with the absorption peaks at the appropriate
m/z ratios.
 Figure 8. Mass Spectrum of Water

Instrumentation
One of the most common types of mass analyzer in GC/MS is the quadrupole ion-trap
analyzer, which allows gaseous anions or cations to be held for long periods of time by
electric and magnetic fields. A simple quadrupole ion-trap consists of a hollow ring
electrode with two grounded end-cap electrodes as seen in figure #. Ions are allowed into
the cavity through a grid in the upper end cap. A variable radio-frequency is applied to the
ring electrode and ions with an appropriate m/z value orbit around the cavity. As the radio-
frequency is increased linearly, ions of a stable m/z value are ejected by mass-selective
ejection in order of mass. Ions that are too heavy or too light are destabilized and their
charge is neutralized upon collision with the ring electrode wall. Emitted ions then strike an
electron multiplier which converts the detected ions into an electrical signal. This electrical
signal is then picked up by the computer through various programs. As an end result, a
chromatogram is produced representing the m/z ratio versus the abundance of the sample.

GC/MS units are advantageous because they allow for the immediate determination of the
mass of the analyte and can be used to identify the components of incomplete separations.
They are rugged, easy to use and can analyze the sample almost as quickly as it is eluted.
The disadvantages of mass spectrometry detectors are the tendency for samples to
thermally degrade before detection and the end result of obliterating all the sample by
fragmentation.

Fig
ure 9. Schematic of the GC/MS system.

 
 F
igure 10. Arrangement of the poles in Quadrupole and Ion Trap Mass spectrometers

Flame Ionization Detectors

Flame ionization detectors (FID) are the most generally applicable and most widely used
detectors. In a FID, the sample is directed at an air-hydrogen flame after exiting the
column. At the high temperature of the air-hydrogen flame, the sample undergoes
pyrolysis, or chemical decomposition through intense heating. Pyrolized hydrocarbons
release ions and electrons that carry current. A high-impedance picoammeter measures this
current to monitor the sample's elution.

It is advantageous to use FID because the detector is unaffected by flow rate,

noncombustible gases and water. These properties allow FID high sensitivity and low
noise. The unit is both reliable and relatively easy to use. However, this technique does
require flammable gas and also destroys the sample.
 Figure 11. Schematic of a typical flame ionization detector.

Thermal Conductivity Detectors

Thermal conductivity detectors (TCD) were one the earliest detectors developed for use
with gas chromatography. The TCD works by measuring the change in carrier gas thermal
conductivity caused by the presence of the sample, which has a different thermal
conductivity from that of the carrier gas. Their design is relatively simple, and consists of
an electrically heated source that is maintained at constant power. The temperature of the
source depends upon the thermal conductivities of the surrounding gases. The source is
usually a thin wire made of platinum, gold or . The resistance within the wire depends upon
temperature, which is dependent upon the thermal conductivity of the gas.

TCDs usually employ two detectors, one of which is used as the reference for the carrier
gas and the other which monitors the thermal conductivity of the carrier gas and sample
mixture. Carrier gases such as helium and hydrogen has very high thermal conductivities so
the addition of even a small amount of sample is readily detected.

The advantages of TCDs are the ease and simplicity of use, the devices' broad application
to inorganic and organic compounds, and the ability of the analyte to be collected after
separation and detection. The greatest drawback of the TCD is the low sensitivity of the
instrument in relation to other detection methods, in addition to flow rate and concentration
dependency.
 Figure 12. Schematic of thermal conductivity detection cell.

Figure 13. Standard Chromatogram of a Mixture of Gases

Chromatogram

Figure 13 represents a standard chromatogram produced by a TCD detector. In a standard

chromatogram regardless of the type detector, the x-axis is the time and the y-axis is the
abundance or the absorbance. From these chromatograms, retention times and the peak
heights are determined and used to further investigate the chemical properties or the
abundance of the samples.
Electron-capture Detectors

Electron-capture detectors (ECD) are highly selective detectors commonly used for
detecting environmental samples as the device selectively detects organic compounds with
moieties such as halogens, peroxides, quinones and nitro groups and gives little to no
response for all other compounds. Therefore, this method is best suited in applications
where traces quantities of chemicals such as pesticides are to be detected and other
chromatographic methods are unfeasible.

The simplest form of ECD involves gaseous electrons from a radioactive ? emitter in an
electric field. As the analyte leaves the GC column, it is passed over this ? emitter, which
typically consists of nickle-63 or tritium. The electrons from the ? emitter ionize the
nitrogen carrier gas and cause it to release a burst of electrons. In the absence of organic
compounds, a constant standing current is maintained between two electrodes. With the
addition of organic compounds with electronegative functional groups, the current
decreases significantly as the functional groups capture the electrons.

The advantages of ECDs are the high selectivity and sensitivity towards certain organic
species with electronegative functional groups. However, the detector has a limited signal
range and is potentially dangerous owing to its radioactivity. In addition, the signal-to-noise
ratio is limited by radioactive decay and the presence of O2 within the detector.

Figure 14. Schematic of an electron-capture detector.

Atomic Emission Detectors

Atomic emission detectors (AED), one of the newest addition to the gas chromatographer's
arsenal, are element-selective detectors that utilize plasma, which is a partially ionized gas,
to atomize all of the elements of a sample and excite their characteristic atomic emission
spectra. AED is an extremely powerful alternative that has a wider applicability due to its
based on the detection of atomic emissions.There are three ways of generating plasma:
microwave-induced plasma (MIP), inductively coupled plasma (ICP) or direct current
plasma (DCP). MIP is the most commonly employed form and is used with a positionable
diode array to simultaneously monitor the atomic emission spectra of several elements.

Instrumentation

The components of the Atomic emission detectors include 1) an interface for the incoming
capillary GC column to induce plasma chamber,2) a microwave chamber, 3) a cooling
system, 4) a diffration grating that associated optics, and 5) a position adjustable
photodiode array interfaced to a computer.

Figure 15. Schematic of atomic emission detector.

GC Chemiluminescence Detectors

Chemiluminescence spectroscopy (CS) is a process in which both qualitative and

quantitative properties can be be determined using the optical emission from excited
chemical species. It is very similar to AES, but the difference is that it utilizes the light
emitted from the energized molecules rather than just excited molecules. Moreover,
chemiluminescence can occur in either the solution or gas phase whereas AES is designed
for gaseous phases. The light source for chemiluminescence comes from the reactions of
the chemicals such that it produces light energy as a product. This light band is used instead
of a separate source of light such as a light beam.

Like other methods, CS also has its limitations and the major limitation to the detection
limits of CS concerns with the use of a photomultiplier tube (PMT). A PMT requires a dark
current in it to detect the light emitted from the analyte.
 Figure 16. Schematic of a GC Chemiluminescence Detector

Photoionization Detectors

Another different kind of detector for GC is the photoionization detector which utilizes the
properties of chemiluminescence spectroscopy. Photoionization detector (PID) is a portable
vapor and gas detector that has selective determination of aromatic hydrocarbons, organo-
heteroatom, inorganice species and other organic compounds. PID comprise of an ultrviolet
lamp to emit photons that are absorbed by the compounds in an ionization chamber exiting
from a GC column. Small fraction of the analyte molecules are actually ionized,
nondestructive, allowing confirmation analytical results through other detectors. In
addition, PIDs are available in portable hand-held models and in a number of lamp
configurations. Results are almost immediate. PID is used commonly to detect VOCs in
soil, sediment, air and water, which is often used to detect contaminants in ambient air and
soil. The disavantage of PID is unable to detect certain hydrocarbon that has low molecular
weight, such as methane and ethane.

Instrumentation

Figure 17. Schematic of a photoionization detector

Limitations

1. Not suitable for detecting semi-volatile compounds

2. Only indicates if volatile organic compounds are presents.
3. High concentration so methane are required for higher performance.
4. Frequent calibration are required.
5. Units of parts per million range
6. Enviromental distraction, especially water vapor.
7. Strong electrical fieldsRapid variation in temperature at the detector and naturally
occurring compounds may affect instrumental signal.

Applications
Gas chromatography is a physical separation method in where volatile mixtures are
separated. It can be used in many different fields such as pharmaceuticals, cosmetics and
even environmental toxins. Since the samples have to be volatile, human breathe, blood,
saliva and other secretions containing large amounts of organic volatiles can be easily
analyzed using GC. Knowing the amount of which compound is in a given sample gives a
huge advantage in studying the effects of human health and of the environment as well.

Air samples can be analyzed using GC. Most of the time, air quality control units use GC
coupled with FID in order to determine the components of a given air sample. Although
other detectors are useful as well, FID is the most appropriate because of its sensitivity and
resolution and also because it can detect very small molecules as well.

GC/MS is also another useful method which can determine the components of a given
mixture using the retention times and the abundance of the samples. This method be
applied to many pharmaceutical applications such as identifying the amount of chemicals in
drugs. Moreover, cosmetic manufacturers also use this method to effectively measure how
much of each chemical is used for their products.

Equations
“Height equivalent to a theoretical plate” (HETP) use to calculate the flow rate by usingthe
total number of theoretical plates (N) and column length (L). Some application, HETP
concepts is used in industrial practice to convert number of theoretical plates to packing
height. HETP can be calculate with the Van Deemter equation, which is given by

Where A and B and C are constants and v is the linear velocity (carrier flow rate).
 A is the "Eddy-Diffusion" term and causes the broadening of the solute band.
  B is the "Longitudinal diffusion" term whereby the concentration of the analyte, in
which diffuses out from the center to the edges.This causes the broadering of the
analyte band.
 C is the "Resistance to Mass Transfer " term and causes the band of the analyte
broader.

L is the length of the column, where N is the number of theoretical plates, tR is the
retention time, and ω is the width of the elution peak at its base.

In which, the more plates give a better resolution and more efficiency. Resolution can be
determined by
 

A relationship between the plates and resolution is giving by,

Where the selectivity, a, and k' is the capacity factors take places of the two solutes. The
selectivity and capacity factors can be control by improving separation, such as changing
mobile/ stationary phase composition, column temperature and use a special chemical
effect.
References
1. Skoog, D. A.; Holler, F. J.; Crouch, S. R. Principles of Instrumental Analysis. Sixth
Edition, Thomson Brooks/Cole, USA, 2007.
2. Krugers, J. Instrumentation in Gas Chromatography. Centrex Publishing Company-
Eindhoven, Netherlands, 1968.
3. Hubschmann, H. Handbook of GC/MS: Fundamentals and Applications. Wiley-
VCH Verlag, Germany, 2001.
4. Scott, R. P. W. Chromatographic Detectors: Design, Function, and Operation.
Marcel Dekker, Inc., USA, 1996.
5. J.N. Driscoll. REview of Photoionization Detection in Gas Chromatography: The
first Decade. Journal of CHromatographic Science , Vol 23. November 1985. 488-
492.
6. Boer, H. , "Vapour phase Chromatography", ed. Desty, D. H., 169 (Butterworths
Sci. Pub., London, 1957).
7. Dimbat, M. , Porter, P. E. , and Stross, F. H. , Anal. Chem., 28, 290 (1956). | Article
| ISI | ChemPort |