International Journal of Agriculture Innovations and Research

Volume 6, Issue 1, ISSN (Online) 2319-1473

Manuscript Processing Details (dd/mm/yyyy) :
Received : 10/07/2017 | Accepted on : 17/07/2017 | Published : 26/07/2017

Gas Chromatography: Principles, Advantages and

Applications in Food Analysis
Wedad Q. AL-Bukhaiti1*, Anwar Noman1, 2 Aseela Saeed Qasim3, Ammar AL-Farga4
1State key Laboratory of Food Science and Technology & School of Food Science and Technology, Jiangnan University, 1800 Lihu,
Avenue, Wuxi 214122, P.R. China.
2Department of Agricultural Engineering, Faculty of Agriculture, University of Sana`a, Sana`a, Yemen
3Department of Chemistry, Faculty of Education, University of Sana`a, Sana`a, Yemen
4School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, PR China

*Corresponding author: Tel: +86 15251516277, E-mail address: wedadqasim@yahoo.com

Abstract – Gas chromatography (GC) is a common kind of
chromatography used as a piece of analytical science for
segregating and investigating exacerbates that can be
vaporized without disintegration. Regular employments of
GC are trying the immaculateness of a particular substance,
or separating of the distinctive segments of a blend.
Development of the analytical methods for identification,
purity evaluation and quantification of drugs and food has
received a great deal of attention in the field of separation
science. This review describes GC method development and
validation in general way. A general and very simple approach
for the GC method development for the separation of
compounds was discussed. Knowledge of the physiochemical
properties of the primary compound is of utmost importance
prior to the any GC method development. Several steps are
being considered for GC method development like column
section (stationary phase and dimensions: column id, length,
and film thickness), carrier gas selection (Nitrogen, Helium,
flow rate), temperature programing (Initial temperature,
initial hold, ramp rate, final temperature, and final hold),
injector selection, Injector temperature, detector selection and
detector temperature. Optimized method is also need to be
validated with various parameters (e.g. specificity, precision,
accuracy, detection limit, linearity, etc.).
Keywords – Gas Chromatography, Development, Column,
Detectors, Application, Validation.
I. INTRODUCTION
Chromatographic separation methods are without any
doubt the most frequently employed analytical techniques
for compositional analysis [1]. Gas chromatography is a
unique and versatile technique. In its initial stages of
development it was applied to the analysis of gases and
vapors from very volatile components. Gas
chromatography is the analytical technique used for
product identification (under very controlled conditions)
and must be directly coupled to a mass spectrometer when
information other than a comparative fingerprint (program)
is required, such as positive identification of peaks on the
chromatogram [2]. The basic principal of gas
chromatography is that greater the affinity of the compound
for the stationary phase, more the compound will be
retained by the column and longer it will be before it is
eluted and detected. Thus the heart of the gas
chromatograph is the column in which separation of the
component takes place, and to this must be added the source
and control of the carrier gas flow through the column, a
mean of sample introduction and a means of detection of
Copyright © 2017 IJAIR, All right reserved
123
the components as they elute from the end of the column.
Since temperature will influence the volatility of the
analytes, the column is placed in a thermostatically
controlled oven [3]. Faster gas chromatographic separation
is a generally beneficial option. Since the decrease time of
analysis results in the increased sample. Reduction of
analysis time can be achieved by changing column
parameters: shorter length, smaller column inner diameter,
thinner film of stationary phase, or operational parameters:
faster temperature program rate, isothermal analysis.
Different carrier gas, higher carrier gas flow rate or a
combination of both approaches can be applied [4].
Chromatography (GC) 50 years ago, GC has been used to
help determine food composition, discover our nutritional
needs, improve food quality, and introduce novel foods.
Furthermore, GC has been the only adequate approach to
measure many of the organic contaminants that occur at
trace concentrations in complex food and environmental
samples. GC has been instrumental in helping humans
realize that we must use caution with agricultural and
industrial chemicals to avoid health and environmental
damage.
Gas Chromatography (GC) is a normally utilized analytic
technique as a part of numerous research and industrial
research facilities for quality control and in addition
identification and quantitation of components in a mixture.
GC is likewise utilized technique as a part of numerous
environmental and forensic labs since it takes into
consideration the detection of very little quantities. An
expansive variety of tests can be analysed the until the
compounds are adequately thermally steady and reasonably
volatile [5].
II. PRINCIPLES OF GAS CHROMATOGRAPHY
The basis of the separation is a retardation of the
individual components as they are moved through a long
column by a carrier gas, usually helium or nitrogen. The
column consists of a steel or glass tube filled with an inert
packing material such as glass or ceramic beads (see Figure
1). In gas-liquid chromatography (GLC), these are coated
with an in volatile liquid, so that the surface area of the
liquid in contact with the gas is large. For some
applications, the packing may be a solid without any liquid
coating; it is then called gas-solid chromatography (GSC),
but this is less widely used than GLC.
The sample is injected into the carrier gas stream. As it
moves through the column with the carrier gas, the
 International Journal of Agriculture Innovations and Research
molecules of each substance present in the sample will
distribute between the gas and the liquid. Individual
molecules will constantly move between the gas and the
liquid in a dynamic equilibrium. While a molecule is in the
gas phase it will pass along the column, while it remains
dissolved in the liquid it will be stationary. The more
volatile a substance, the greater proportion of time its
molecules will be moving in the carrier gas, and so the
sooner it will emerge from the column. In this way each
substance will become separated within the column and
emerge separated by time at the end.
The time taken from injection to emergence is known as
the retention time (Rt), and is characteristic for each
substance under any given set of conditions. It depends on
the volatility of the substance, as well as the temperature of
the column and its length and diameter. Many substances
have an inconveniently long retention time at room
temperature, and this is overcome by heating the column in
an oven. Having separated the components in the column
so that they emerge individually, some method of detecting
and measuring them is needed. Two types of detector are
commonly used: thermal conductivity and flame ionization.
Fig. 1. Schematic Diagram of Gas Chromatography
III. APPLICATIONS
Gas chromatography (GC) is used widely in applications
involving food analysis. Typical applications pertain to the
quantitative and/or qualitative analysis of food
composition, natural products, food additives, flavor and
aroma components, a variety of transformation products,
and contaminants, such as pesticides, fumigants,
environmental pollutants, natural toxins, veterinary drugs,
and packaging materials [7]. And particular food
applications involving GC, such as carbohydrates and
amino acids [8]. Lipids and accompanying lipophilic
compounds [9, 10]. flavors and aroma [11, 12].
GC can be used for the direct separation and analysis of
gaseous samples, liquid solutions, and volatile solids. If the
sample to be analyzed is non-volatile, the techniques of
derivatization or pyrolysis GC can be utilized [2]. Gas
Copyright © 2017 IJAIR, All right reserved
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Volume 6, Issue 1, ISSN (Online) 2319-1473
Thermal conductivity detectors (TCDs) rely on changes in
the thermal conductivity of the gas leaving the column. The
pure helium carrier gas passes over a hot tungsten-rhenium
filament, causing it to cool, since helium has a very high
thermal conductivity. When a chemical substance emerges
with the carrier gas, cooling will be less, and the
temperature of the filament will rise. As with most metals,
its electrical resistance increases with temperature and this
can be measured and recorded. Flame ionization detection
(FID) is more frequently encountered in food applications,
since many compounds under investigation are organic
(containing carbon), and FID is around a thousand times
more sensitive than thermal conductivity detection for
organics. The gas emerging from the column is burned with
a hydrogen and air mixture. This forms ions, which conduct
an electric current which can be amplified and recorded on
a chart recorder. Although the number of ions formed in
this way is small, perhaps only 0.0001 per cent of the total
carbon atoms present in the sample, the proportion
produced is always constant. This means that the total
signal recorded on the chart recorder is proportional to the
amount of the chemical substance present [6].
chromatography (GC) has been an indispensable analytical
technique in the application of fatty acid determinations in
oilseed plant breeding, biosynthesis and human
metabolism. As well as the characterization of complex
mixtures of geometric isomers when combined with other
chromatographic separations and spectroscopic
identification [13]. Plant breeders uses GC as a more precise
and rapid technique for studying the variation and
inheritance of fatty acids in oilseed crops such as rapeseed
[14]. flaxseed [15], and safflower [16].
IV. ADVANTAGES OF GC PERFORMANCE
Optimum qualitative and quantitative GC analysis of
complex mixtures presupposes: (1) good resolution, as
shown by sharp and symmetric peaks; (2) high repeatability
and reproducibility of retention times; (3) high precision
 International Journal of Agriculture Innovations and Research
and accuracy in quantitation based on peak area
measurements, i.e. no discrimination of components
through volatility, polarity or concentration; (4) minimum
thermal and catalytic decomposition of sensitive sample
components [17]. The use of fused-silica capillary columns
with improved surface inertness, thermal stability and
resolution [18]. Best fulfils most of these requirements. In
capillary GC the peak resolution, expressed in terms of
column efficiency, separation and retention factors [19]. Is
primarily affected by the polarity of the stationary phase,
column length, internal diameter and film thickness [20, 21,
22]. A variety of columns with different properties are
available. In addition, fused-silica columns are highly
applicable in practical work due to their flexibility and
simplicity in handling and easy connection to GC and mass
spectrometers [23]. In order to improve the sensitivity of the
GC analyses it is also important to test the effect of carrier
gas flow rate, as well as gas flows in the FID, in order to
reduce the noise from the hydrogen flame [24]. The carrier
gas, usually hydrogen or helium, and its purity can also
affect the resolution [20].
V. DEVELOPMENT OF GC METHOD
Methods are developed for new products when no official
methods are available. Alternate method for existing (Non-
Pharmacopoeial) products are to reduce the cost and time
for better precision and ruggedness. When alternate method
proposed is intended to replace the existing procedure
comparative laboratory data including merit/demerits are
made available [25]. Several steps are being considered for
GC method development like column selection (stationary
phase and dimensions: column id, length, and film
thickness), carrier gas selection (Nitrogen, Helium, flow
rate), temperature programing (Initial temperature, initial
hold, ramp rate, final temperature, and final hold), injector
temperature and detector temperature [26].
Steps involved in Method development [27]:
- Understanding the Physicochemical properties of sample.
- Selection of chromatographic conditions.
- Developing the approach of analysis.
- Sample preparation.
- Method optimization.
- Method validation.
VI. SELECTION OF CHROMATOGRAPHIC
CONDITIONS
1. Column Selection
A column is of course, the starting and central piece of a
chromatograph. An appropriately selected column can
produce a good chromatographic separation which provides
an accurate and reliable analysis. An improperly used
column can often generate confusing, inadequate, and poor
separations which can lead to results that are invalid or 1-2 Sample Injection Port
complex to interpret. There are over 10, 000 compounds
that can be analysed by GC and over 400 GC capillary
columns [26]. The initial choices of column and supporting
instrumentation have a profound influence on the
possibilities for, and ultimate results of, separation
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Volume 6, Issue 1, ISSN (Online) 2319-1473
optimization. Manipulation of column parameters
(stationary phase, inner diameter, length, and film
thickness) gives chromatographers control over column
efficiency, resolution and speed of analysis. [2]. In gas
chromatography, the elution order of analytes is governed
by several factors such as latent vapour pressures,
solubilities in stationary phase, and propensities for
molecular interaction in the stationary phase. All of these
effects change with temperature and their concerted effect
ultimately determine the equilibrium distribution of solute
molecules between the mobile and stationary phase [28].
Selection of Column:
A column is of course, the starting and central piece of a
chromatograph. An appropriately selected column can
produce a good chromatographic separation which provides
an accurate and reliable analysis. An improperly used
column can often generate confusion, inadequate, and poor
separations which can lead to results that are invalid or
complex to interpret. There are over 10, 000 compounds
that can be analysed by GC and over 400 GC capillary
columns. It is a challenge for a column manufacturer to give
detailed column selection guidelines to meet such a wide
variety of applications. An optimized chromatographic
separation begins with the column. The selection of the
proper capillary column for any application should be based
on four significant factors which are: stationary phase,
column internal diameter, film thickness, and column
length. The differences in the chemical and physical
properties of injected organic compounds and their
interactions with stationary phase are the basis of separation
process. When strength of the analyte-phase interactions
differs significantly for two compounds, one is retained
longer than the other. How long they are retained in the
column (retention time) is a measure of these analyte-phase
interactions. Changing the chemical features of the
stationary phase alters its physical properties. Two
compounds that co-elute (do not separate) on a particular
stationary phase might separate on another phase of
different chemistry [26].
1-1 Selection of Column Internal Diameter (i.d.) High
Efficiency
Column id plays two contradicting roles in separation.
The efficiency of a capillary column, measured in plates (n)
or plates per meter (n/m), increases as the i.d. of the column
decreases. This is one of the basic principles behind fast GC
but decreased sample loading capacities. When a column is
overloaded with sample, the plate number is decreased
greatly. If the sample to be analysed contains many
analytes, or has analytes that elute closely together, the most
narrow i.d. capillary column that is practical should be
selected. Note that very narrow bore columns, such as 0.10
or 0.18 mm i.d., may require specialized equipment, such
as GC with a pressure regulator that allows higher column
head pressure [29].
For optimum column potency, the sample mustn't be
large, and may be introduced onto the column as a "plug"
of vapor - slow injection of enormous samples causes band
broadening and loss of resolution. The foremost common
injection methodology is wherever a micro syringe is
 International Journal of Agriculture Innovations and Research
employed to inject sample through a rubber septum into a
flash vaporizer port at the top of the column. The
temperature of the sample port is sometimes concerning
50oc beyond the boiling purpose of the smallest amount
volatile element of the sample [30-32].
1-3 Column Temperature
For precise work, column temperature should be
controlled to at intervals tenths of a degree. The optimum
column temperature is dependent upon the boiling purpose
of the sample. As a rule of thumb [33], a temperature
slightly on top of the typical boiling purpose of the sample
ends up in associate extraction time of two - half-hour.
Least temperatures offer sensible resolution, however
increase extraction times [34].
2. Selection of Carrier Gas:
Finding the Best Carrier Gas Average Linear Velocity
Determining the best average linear velocity is fairly easy
and only involves a small amount of trial and error.
Hydrogen provides the best resolution in shortest amount of
time. Helium provides similar resolution, but at a longer
analysis time. Nitrogen is not recommended for use with
capillary columns due to the extremely long analysis times.
When using helium as the carrier gas, try an initial average
linear velocity of 30 cm/sec. If better resolution is desired,
reduce the velocity to not less than 25 cm/sec; however, the
analysis time will be increased. If a shorter analysis time is
desired, increase the velocity to 35-40 cm/sec; be aware of
potential resolution losses at these higher linear velocities.
Average linear velocities of 30-35 cm/sec are used for many
analyses when using helium as a carrier gas. When using
hydrogen as the carrier gas, try an initial average linear
velocity of 60 cm/sec. If better resolution is desired, reduce
the velocity to not less than 50 cm/sec; however, the
analysis time will be increased. If a shorter analysis time is
desired, increase the velocity to 70-80 cm/sec; be aware of
potential resolution losses at these higher linear velocities.
Average linear velocities of 60-70 cm/sec are used for many
analyses when using hydrogen as carrier gas. [35]. The
choice of gas to be used as mobile phase in gas
chromatography is influenced by the following
requirements and considerations:
Inertness, Dryness, Freedom from oxygen, Safety, Cost
and Availability [3].
3. Optimization of Column Oven Temperature
Program.
The column resides in an oven, and temperature, which
greatly affects the effectiveness of the chromatographic
separation, is an extremely important factor used in
controlling GC. In many cases, isothermal is not the most
effective temperature mode for sample separation; in such
cases, a temperature program can be used. Most GC
temperature program have initial temperature, a ramp
(degree increase per minute) and a final temperature. Using
a linear temperature program as a starting point if previous
analysis information is not available to use as a guide, the
first program development step is to try a simple, linear
temperature program. To improve the resolution of earlier
eluting peaks, decrease the initial temperature or increase
the initial hold time. Decreasing the initial temperature
usually results in the largest resolution improvement, but
Copyright © 2017 IJAIR, All right reserved
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Volume 6, Issue 1, ISSN (Online) 2319-1473
analysis times are substantially increased. The resolution of
peaks eluting in the middle of the chromatogram can be
altered by change in ramp rate. If there is excessive peak
resolution, the ramp rate can be increased to reduce
resolution and the analysis time. If there is insufficient
resolution, decrease the ramp rate, but there will be an
increase in the analysis time. Better resolution of later
eluting peaks often occurs when decreasing the ramp rate.
Another option to alter resolution of peaks in the middle of
a chromatogram is to use a mid-ramp hold. A mid ramp hold
is a several minute isothermal portion somewhere during a
temperature ramp. Stop the temperature program shortly
after last peak has eluted from the column. If column’s
isothermal temperature limit is reached and peaks are still
eluting, a final hold time is necessary. Only use a final hold
time if the temperature limit is reached [26, 35].
4. Optimization of Injector Type, Temperature &
Injection Volume
Introduction of the sample into GC system is a critical
step in separation. The reproducibility of the amount of
sample injected is important to ensure the reproducibility of
results. A sample can be injected manually into the system
or by using an auto sampler system. A major errors in GC
is poor injection technique. The injector temperature for
separation. The temperature of injector is used to rapidly
vaporize the liquid sample into gaseous phase that can be
carried to the column for separation. In capillary and micro
packed gas chromatography (GC), there are four primary
techniques for vaporizing a sample and transferring it onto
the inlet of the analytical column: split, splitless, direct, and
on-column injections. Of these, split and splitless injections
are the most commonly used techniques. Split Injector was
selected for analysis of sample with high concentration
levels. In the split injection mode, only a fraction of the
vaporized sample is transferred onto the head of the
column. The remainder of the vaporized sample is removed
from the injection port via the split vent line. Split injections
should be used only when sample concentrations are high
enough to allow a portion of the sample to be discarded
during the injection process, while still maintaining a
sufficient concentration of analytes at the detector to
produce a signal [35, 36].
5. Selection of Stationary Phase
When selecting a column, first determine the samples
characteristics to match the columns stationary phase.
Stationary phases are in general divided into 3 categories
first non-polar second mid-polar and third Polar. Stationary
phases are further categorized by Siloxane (non-polar and
mid-polar) and Polyethylene glycol (PEG or polar). G1, G2
and G38 (100% Methyl Polysiloxane) does not undergo
hydrogen bonding interactions. The change in the elution
order of Hexanol and Phenol with G14, G15, G16, G20,
G39 and G47 (Polyethylene glycol) is a combination of
dipole and hydrogen bonding interaction [26].
6. Optimization of Detector Type & Detector
Temperature
A variety of detector is commercially available to be used
with GC, each having its own limitations and advantages.
The most commonly used detector in GC is flame ionization
detector. Detector temperatures and the relative flow rates
 International Journal of Agriculture Innovations and Research
Volume 6, Issue 1, ISSN (Online) 2319-1473

of carrier gas, hydrogen and air into the detector are the key
operating parameters [26]. A series of standards is defined
for evaluation of detector parameters such as drift, noise,
sensitivity, linear range, dynamic range etc. The variation
in detector response with flow rate depends on whether the
detector is concentration or mass flow dependent. For
concentration dependent detectors (e.g., thermal
conductivity detector, photo ionisation detector) a decrease
in the flow-rate does not affect the peak height, which
remains approximately constant. However the peak width,
and consequently the peak area, increase. In contrast, for
mass flow detection systems (e.g., flame ionization
detection, flame photometric detection, nitrogen
phosphorus detection) the response is inversely
proportional to the retention time [37].
7. Detectors
There are several detectors which may be employed in
gas activity. Totally different detectors can offer differing
kinds of property. The response of a mass flow dependent
detector is unaffected by make-up gas. As shown in the
table (1) [38-43].

Table: 1 Types of detectors which may be employed in gas activity

Detector Type Support gases Selectivity Detectab Dynamic
ility range
Flame ionization Mass flow Hydrogen and Most organic cpds. 100 pg 107
(FID) air
Thermal conductivity Concentration Reference Universal 1 ng 107
(TCD)
Electron capture Concentration Make-up Halides, nitrates, nitriles, peroxides, 50 fg 105
(ECD) anhydrides, organometallics
Nitrogen- phosphorus Mass flow Hydrogen and Nitrogen, phosphorus 10 pg 106
air
Flame photometric Mass flow Hydrogen and Sulphur, phosphorus, tin, boron, arsenic, 100 pg 103
(FPD) air possibly germanium, selenium, chromium
oxygen
Aliphatics, aromatics, ketones, esters, 2 pg 107
Photo- ionization Concentration Make-up aldehydes, amines, heterocyclics,
(PID) organosulphurs, some organometallics

The effluent from the column is mixed with gas and air,
and lit. Organic compounds burning within the flame
manufacture ions and electrons which might conduct
electricity through the flame [44]. An outsized electrical
potential is applied at the burner tip, and a collector
conductor is found higher than the flame [45]. The present
ensuing from the transformation of any organic compounds
is measured [46].
VII. CONCLUSION
Gas chromatography is an important analytical technique
for qualitative and quantitative analysis in a wide range of
application areas. It is fast, provides a high peak capacity,
is sensitive and allows combination with a wide range of
selective detection methods including mass spectrometry.
However, the application area of GC is limited because the
molecules to be analysed have to be thermally stable and
sufficiently volatile. Numerous molecules do not meet these
requirements and hence are not amenable to direct GC
analysis. Recent research has resulted in better
chromatographic columns and methods for sample
preparation that enable a significant expansion of the
molecular application range of GC. The strategies exploited
include conversion of (macro) molecules into smaller
species and approaches to reduce the polarity of molecules.
Abbreviations
mm millimeter
fg femtogram
ng nanogram
pg pictogram
Copyright © 2017 IJAIR, All right reserved
127
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AUTHORS' PROFILES
First Author:
Wedad Q. AL-Bukhaiti
Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, Jiangsu, P.R.
China.
Second Author:
Anwar Noman
School of Food Science and Technology, Jiangnan University, 1800 Lihu
Avenue, Wuxi, 214122, Jiangsu, P.R. China.