GAS

CHROMATOGRAPHY
Submitted to : Dr. D. Sindhanaiselvi

Submitted by : Kumar Gaurav

Reg. No. : 19CE1032
Section : CE1
Department : Civil Engineering
Contents:
 Introduction
 Principle
 Advantages
 Instrumentation
 Working
 Evaluation
 Application
 Reference
Introduction:

• Gas chromatography – It is a process of separating component(s)

from the given crude drug by using a gaseous mobile phase.

• It involves a sample being vaporized and injected onto the head of

the chromatographic column. The sample is transported through
the column by the flow of inert, gaseous mobile phase. The column
itself contains a liquid stationary phase which is adsorbed onto the
surface of an inert solid.
Two Major Types of Gas Chromatography:

• Gas-Solid Chromatography:
In this, the mobile phase is a gas while the stationary phase is a
solid.
It is used for separation of low molecular gases, e.g., air
components, H2S, CS2 ,CO2, rare gases, CO and oxides of
nitrogen .

• Gas-Liquid Chromatography:
The mobile phase is a gas while the stationary phase is a liquid
retained on the surface as an inert solid by adsorption or chemical
bonding.
Principles:
• The principle of separation in GC is “partition.”
• The mixture of component to be separated is converted to vapour and
mixed with gaseous mobile phase.
• The component which is more soluble in stationary phase travel slower
and eluted later. The component which is less soluble in stationary phase
travels faster and eluted out first.
• No two components has same partition coefficient conditions. So the
components are separated according to their partition coefficient.
• Partition coefficient is “the ratio of solubility of a substance distributed
between two immiscible liquids at a constant temperature.”
Advantages:

 The technique has strong separation power and even complex

mixture can be resolved into constituents.
 The sensitivity of the method is quite high.
 It gives good precision and accuracy.
 The analysis is completed in a short time.
 The cost of instrument is relatively low and its life is generally
long.
 The technique is relatively suitable for routine analysis.
Instrumentation:
• Carrier gas
- He (common), N2, H2, Argon
• Sample injection port
- micro syringe
• Columns
• Detectors:
 Thermal conductivity (TCD)
 Electron capture detector(ECD)
 Flame Ionization detector (FID)
 Flame photometric (FPD)
Schematic Diagram of a Gas Chromatograph:

Source: Google Images

Carrier Gas:
 The carrier gas must be chemically inert.
 Commonly used gases include nitrogen, helium, argon, and carbon
dioxide.
 The choice of carrier gas is often dependant upon the type of detector
which is used.
 The carrier gas system also contains a molecular sieve to remove water
and other impurities.
o P inlet 10-50 psig
o F=25-150 mL/min packed column
o F=1-25 mL/min open tubular column
Sampling unit:-

• Sampling unit or injection port is attached to the

column head. Micro syringe

• Since the sample should be in vaporized state, the injection port is

provided with an oven that helps to maintain its temperature at
about 20-500 C above the boiling point of the sample.
• Gaseous samples may be introduced by use of a gas tight
hypodermic needle of 0.5-10 ml capacity.
• For Liquid samples , micro syringes of 0.1-100μL capacity may be
used.
Columns:
 There are two general types of column, packed and capillary (also known as
open tubular).

 Packed columns contain a finely divided, inert, solid support material

(diatomaceous earth) coated with liquid stationary phase. Most packed
columns are 1.5 - 10m in length and have an internal diameter of 2 - 4mm.

 Capillary columns have an internal diameter of a few tenths of a millimeter.

They can be one of two types; wall-coated open tubular (WCOT) or support-
coated open tubular (SCOT).
• Wall-coated columns consist of a capillary tube whose walls are coated
with liquid stationary phase. In support-coated columns, the inner wall
of the capillary is lined with a thin layer of support material such as
diatomaceous earth, onto which the stationary phase has been adsorbed.
• SCOT columns are generally less efficient than WCOT columns.
Both types of capillary column are more efficient than packed columns.
Detectors:

 The eluted solute particles along with the carrier gas exit from the
column and enter the detector.
 The detector then produces electrical signals proportional to the
concentration of the components of solute.
 The signals are amplified and recorded as peaks at intervals on
the chromatograph.
Properties of an Ideal Detector :

• Sensitive
• Operate at high Temperature T (0-400 C)
• Stable and reproducible
• Linear response
• Wide dynamic range
• Fast response
• Simple and Reliable
• Non-destructive
• Uniform Response to all analytes
1. Thermal Conductivity Detector:

o TCD is based upon the fact that the heat lost from a filament
depends upon the thermal conductivity of the stream of
surrounding gas as well as its specific heat.
o When only carrier gas flows heat loss to metal block is
constant, filament T remains constant.
o When an analyte species flows past the filament generally
thermal conductivity changes, thus resistance changes which is
sensed by Wheatstone bridge arrangement.
o The imbalance between control and sample filament
temperature is measured and a signal is recorded.
• Advantages :
 Simple and inexpensive
 Durable and posses long life
 Accurate results
 Non-selective, hence known as universal detectors

• Disadvantages:
 Low sensitivity
 Affected by fluctuations in temperature and flow
rate.
2. Electron capture detector :
 Molecules of compounds, which posses affinity for electrons, differ in their
electron absorbing capacities. This difference is utilized in this detector for
identification of the compounds.
 Working - A foil made up of a radioactive metal like Ni63 (β- emitter) is
placed inside a Teflon coated cell which also contains a cathode and an anode.
 In the absence of organic species, the produced electrons migrate towards
positive electrode and produce a certain constant standing current.
 When a sample/eluent is present it captures the electrons, elutes from column,
there is a drop in this constant current.
 The potential across two electrodes is adjusted to collect all the ions and a
steady saturation current, is therefore, recorded.
 Advantages:-
• Highly selective
• Highly sensitive for the detection of compounds like halogens,
quinones, peroxides, nitrites, etc.
• It is non-destructive
• More sensitive than TCD and FID.

 Disadvantages:-
• Least sensitive to compounds whose molecules have negligible
affinity for electrons.
• Carrier gas used should be of pure form like pure nitrogen.
3. Flame ionization detector:

 This employs hydrogen flame that is maintained in a small cylindrical jet

made up of platinum or quartz.
 Effluent from the column with helium or nitrogen as carrier gas are fed
into the hydrogen flame, gets ignited and undergoes pyrolysis to produce
ions.
 For detection of these ions, two electrodes are used that provide a
potential difference.
 The ions produced are repelled by the positive electrode which hit the
collector plate. The current produced in doing so is amplified and fed to
an appropriate recorder.
4. Flame photometric detector:

 It is a selective detector that is responsive to compounds containing

sulphur or phosphorous.
 The detection principle is the formation of excited sulphur (S2*) and
excited hydrogen phosphorous oxide species (HPO*) in a reducing
flame.
 A photomultiplier tube measures the characteristic chemiluminescent
emission from these species. The optical filter can be changed to allow
the photomultiplier to view light of 394 nm for sulphur measurement or
526 nm for phosphorus.
 Applications:
o For detection of heavy metals like
chromium, selenium, tin, etc, in
organometallic compounds.
o Also for analysis of pesticides, coal,
hydrogenated products as well as Flame photometric detector
air and water pollutants.
Working:
 Fill the syringe with sample.
 Record the setting i.e., column temperature, detector temperature and injection port
temperature.
 Introduce sample into the injection port by completely inserting the needle into the
rubber septum. Note down the injection time.
 The sample gets vapourized due to higher temperature of injection port and is swept
into column by carrier gas.
 This sample components now get distributed between the gas and stationary liquid
phase depending upon their solubilizing tendencies.
 The components with minimal solubility move faster and those with maximum
solubility travel slowly.
 The components leaving the column activate detector and recorder to give a plot.
 The components that slowly leave the column give a bell
shaped curve of shorter peak while the one which travels faster
gives a bluntly pointed curve of larger peak.
 In above graph, the component that first emerges out of the
column is component 4 followed by 2,5,3 and 1.
 The area under the curve is determined in order to obtain the
percentage composition of the mixture.
Evaluation:
 HETP (height equivalent to theoretical plate)- It is the distance on the column in
which equilibrium is attained between the solute in the gas phase and the solute in
liquid phase. Larger the number of theoretical plates/ smaller the HETP, the more
efficient the column is for separation.
HETP = Length of column/n ; Where n = number of theoretical plates
 Retention Time: defined as the absolute time taken by a sample to show maximum
peak after injecting.
 Retention Volume: defined as the volume of gas required to elute about half of the
solute through the column.
VR = tR ×F
F = average volumetric flow rate (mL/min), estimated by using soap bubble
meter (some gases dissolving in soap solution).
Application:
 Qualitative Analysis – by comparing the retention time or volume of the sample to
the standard / by collecting the individual components as they emerge from the
chromatograph and identifying these compounds by other methods like UV, IR,
NMR.
 Quantitative Analysis- area under a single component elution peak is proportional
to the quantity of the detected component/response factor of the detectors. It is
done by:
I. Direct comparison method-
A(sample) / A(std) = α C(sample)/C(std)
Where, α is the response factor determined for every pure compound under
given conditions.
II. Calibration curve- a graph is plotted by taking peak areas on Y axis
and concentration of standard compound on X axis. Concentration of
unknown sample is then determined by plotting its peak area on
same graph.

III. Internal standard method- A known concentration of internal

standard, which has similar retention characteristics as that of
sample is added to both reference standard and test sample.
• Pharmaceutical applications-
 Quality control and analysis of drug products like antibiotics
(penicillin), antivirals (amantidine), general anesthetics (chloroform,
ether), sedatives/hypnotics (barbiturates), etc.
 Assay of drugs – purity of a compound can be determined for drugs
like :
 Atropine sulphate
 Clove oil
 Stearic acid
 In determining the levels of metabolites in body fluids like plasma,
serum, urine, etc.
References:
o Wikipedia
o Google
o Google Images
o Google Slides
o Slideshare
o Library Genesis
o Department of E&I, SIT Tumkur
o Skoog.D.A, Holler .F.J; principles of instrumental analysis.
Thank You