GAS

CHROMAROGRAPHY

By: Madam Nurul Fasihah Razak

What Is
This Topic
About?

What is GC?
What it is used for?
Why it is important to learn?
 LEARNING OUTCOMES
Explain the basic ILLUSTRATE the

01 PRINCIPLE of Gas
Chromatography 03 schematic diagram of
GC instrumentation.
(GC).

Describe the FUNCTION

02 04
Describe the TYPE for each components in GC
GC spectrophotometry and
the applications.
 Terminology
Mobile phase: gas that carries the sample/analyte through the
stationary phase

Stationary phase: where separation take place. It can be almost

anything.

Eluent: mobile phase (gas)

Eluate: species eluted from the bottom of a chromatographic column

Chromatogram : is a graph/ plot obtained from an analysis. Each peak

in the chromatogram represents a single compound.
Retention time :
The time at which the peak reaches its maximum

Absolute retention time:

time from the moment of injection required for a sample to reach the detector

Dead time:
time between injection of a sample and the appearance of the first peak on
the chromatogram

Relative RT :
time relative to some internal standard requires for a sample to be detected.
 Basics principles
 An analytical tech. for separating compounds based primarily on their volatilities.
 Separation occurs based on the interactions of the sample (solute) with the
mobile and stationary phases.
 Provides both qualitative (what is inside) & quantitative (how much) information
for individual compounds present in sample.

 Sample (solute) is dissolved in a solvent eg. Methanol

 Sample and solvent are vaporized onto the head of a column
 Basics principles
 Vaporized solvent and solute are carried through the column by an inert gas
(mobile phase)
Note: the mobile phase does not interact with compounds of interest
 All of the solvent passes through the column (unretained)
Note: the solvent is not the mobile phase
 Separation occurs by interaction of solute with a stationary phase
 Detection occurs by a variety of means (e.g. thermal conductivity, flame
ionization, thermionic or electron capture detectors
 Types of GC
We distinguish three types of gas chromatography based on the type of
stationary phase involved:

GAS-LIQUID GAS-SOLID GAS-BONDED

PHASE

• The stationary phase is • The stationary • The stationary phase

a liquid adsorbed on a phase is a is an organic material
solid surface. solid material bonded to a solid
• most common type. surface
GC- Schematic Diagram
GC- Schematic Diagram

READ OUT
 Basic Components
1. Carrier Gas (mobile phase)
He, Ar, N2 and H2
2. Sample Injection System
3. Chromatographic Column
• Contains stationary phase
• Many configurations
4.Oven- Isothermal or temperature programmed
5.Detector
FID, TC, ECD, MS, etc.
6.Readout
Chromatography software (strip chart and integrator package)
 Fundamentals principles of GC
• is a process by which a mixture of volatile materials
(samples) is separated by adsorption between two
immiscible phases, which is mobile phase and stationary
phase.
• Samples normally organic compound , vaporizable at
400º C.
• Other samples may be amenable by after sample pre-
treatment method
• Column is the heart of GC for separations occurred.
 Column

The vaporized sample is carried through the column by a carrier gas.

Component of column – Tubing and packing/stat. phase.
Tubing is usually made of glass, stainless steel, copper, aluminum, nickel,
glass lined, fused silica.
 Column selection depends on the sample to be analysed.
 Resolving power is increased with longer column
 Analysis time is increased with longer column
 Column temp. – isothermal : RT increase
 However, the general the general rule is LIKE SEPARATE LIKE.
 GC Column Types
1. Packed column

○ Used in early gas-liquid chromatography

○ Typically made of glass, Teflon and aluminum

○ Length typically 2-3 m; ID ~3 mm

○ Filled with a material called a “solid support”; the material which holds the stationary phase

2. Capillary column

○ Wall coated Open Tubular ( WCOT ) – polymer film coated on the inner wall of the column

○ Porous Layer Open Tubular ( PLOT ) – porous solid particles coated on the inner wall of the column

○ Packed capillary column – solid particles packed inside the column

Stationary Phase
Often referred to as the “immobilized liquid phase”
Desirable Properties:
1. Low volatility
2. Thermal Stability
3. Chemical inertness
4. Solvent characteristics that optimize k’ and α
Choice of stationary phase is often made based on polarity
For the solute and stationary phase to interact they must have similar
polarities
“Like dissolves like”
 Sample Injector
● Function: to change sample to a gas & to
introduce the resulting gaseous sample into
column.
● Purpose: to introduce sample as “plug” at
the head of the column
○ Effects band broadening
● Injector typically 50 °C hotter than oven and
higher than boiling point of sample

● Sample is “flash evaporated” and expands

into gas expansion chamber

● Injection volumes are small

○ Capillary columns ~ 1 µL

○ Packed columns 1-20 µL

 Desirable GC injector
qualities:
o Vaporize all solutes instantaneously without thermal
decomposition.
o No sample loss through adsorption
o No sample contamination
o Injection of sample into mobile phase without sample dispersion
or tailing
o Avoid diffusion of sample components in mobile phase
 INJECTOR TYPES

• Split/Splitless injector
• Advantage: prevent introduction of non-volatile components
in the sample
• Disadvantage: discrimination against high boiling compounds
and thermally unstable compounds
• Wide bore injector
• On-column Injector ( OCI )
• Programmed Temperature Vaporizing ( PTV ) Injector
SPLIT SPITLESS INJECTION
for higher concentration samples. during injection, split flow line is
Sample vapor mixed with carrier closed.
gas will flow with the carrier gas.

A small flow goes into the column Almost all of sample vapor goes into
( 1- 4 ml/min typically ). the column.
A much larger flow ( 10- 100 ml/min Split flow line is re-opened after
typically ) goes out from the split about 30 sec to 2 min.
outlet.
Typical injection volume : 1- 2
microlitre.
Advantage : good peak shapes in Good for semi-volatile samples.
general
For trace level compounds.
 Oven

• Isothermal or temperature programmed

• Higher temperature – poor separation, better
shape peaks
• Lower temperature – better separation, poor
shape peaks
 Detectors
• Detector indicates the presence of each component that elutes
out from the column
• FID – for compounds that can be oxidized in hydrogen –air
flame
• FPD , ECD, TCD, FTD
Thermal Conductivity Detector
(TCD)
 Detection Principle: analyte gases have different
thermal conductivities than carrier gases
 A platinum, gold or tungsten wire (or a thermister)
is placed in the exit gas stream from the column
 A constant voltage is applied to heat the wire
 Temperature/Resistance of the wire is proportional
to the thermal conductivity of the surrounding gas
 Double detector system: one detector in carrier
gas and one in the carrier + analyte
 Cancels out resistance due to carrier gas giving
signal only for analyte
Flame Ionization Detector
(FID)
 Organic analytes are pyrolyzed in an
air/H2 flame
 Ions are produced in the plasma
around the flame
 proportional to number of
carbons present
 Positive voltage is applied to
collector; negative to the flame body
 Ions migrate to collector producing a
current (signal)
GC-FID
Thermionic or
Nitrogen/Phosphorus
Detector
 Similar to FID
 Flame flows around an heated
rubidium silicate bead
 Heated bead forms a plasma (600-
800 °C)
 Sensitive to P and N
 Ion current for P & N > 104 times
that of C
 Electron Capture Detector
(ECD)
How it Works
• Column effluent is passed over a ß- emitter
– Tritium or 63-Ni
• The carrier gas is ionized
• A burst of e- is produced with each radioactive decay
• Potential is applied between the collector (anode) and the detector
body (cathode)
• Produces a constant background current
• The current flow decreases in the presence of analyte molecules
– The analyte captures the emitted electrons

Detector Characteristics
• Sensitive to molecules containing electronegative functional
groups (e.g. Cl- )
• Non-linear response to analyte concentration
Photoionization Detector

• An UV source ionizes all the

molecules in the column
effluent
• Ions produced are collected
resulting in a current flow
 Other GC Detectors
• Mass Spectrometer
– GC-MS “hyphenated” system
– Used for identification purposes
• Atomic Emission Detector
– GC-AED
• Sulfur Chemiluminescence Detector (SCD)
• Fourier-Transform Infrared (FTIR)
– Used with polar molecules
 Characteristics of GC Detectors
Gas Chromatography Detectors – Characteristics

Sensitivity
(g solute/mL
Detector Signal Signal of carrier
Type Generator Produced Advantages Disadvantages gas)
Thermal Thermal conductivity of Change in the electrical  Universal detector  Low sensitivity
Conductivity analyte gas (usually less resistance of wire in analyte  Simplicity  Often can’t use with capillary ~ 10-8 g
than that of carrier H2 or stream (measure i & R at  Large linear dynamic range columns
He constant applied voltage; or  Non-destructive (10-100 ppm)
change in temperature when  Sensitive to all compounds
thermister used)
Flame Ionization of analyte in Ion current  Very sensitive  Insensitive to H2O, CO2, SO2,
Ionization H2/air flame  Large linear dynamic range (`107) and NOx ~ 10-13 g
 General purpose detector  Destructive
 Responds to reduced carbon in
analytes
Thermionic Ionization of analyte in Ion current  Selective towards organic  Selective towards organic Similar to the
H2/air flame compounds containing P or N compounds containing P or N FID, but for N
compared to C (~104 to 106 X) compared to C (~104 to 106 X) and P
Electron Reduction in the Ion current; decreases in the  Selective towards organic  Selective towards organic
Capture ionization of a carrier presence of organic molecules compounds containing compounds containing ~ 10-15
gas (e.g. N2) by a which can capture electrons electronegative functional groups (e.g. electronegative functional
radioactive (ß- emitter) halogens, peroxides, quinines, and groups (e.g. halogens,
source, usually 63-Ni nitro groups) peroxides, quinines, and nitro
 Non-destructive groups)
 Very sensitive  Insensitive to amines,
alcohols, hydrocarbons,
 Small linear dynamic range
(~100)
Mass Ionization of analyte Analyte ions; discrimination  Universal detector  Cost
Spectrometer based on mass to charge ratio of  Good for complex organic mixtures  Complexity ~ 10-12
analyte ions  Speed  Ease of use
 High sensitivity
 Compound identification
EXAMPLE OF THE CHROMATOGRAM
 Criteria to optimize separation

● i.Capacity factor
● k', is often used to describe the migration rate of an analyte on a column. You may
also find it called the capacity factor. When an analytes retention factor is less
than one, elution is so fast that accurate determination of the retention time is very
difficult. High capacity factors (greater than 20) mean that elution takes a very long
time. Ideally, the retention factor for an analyte is between one and five.
● When calculating the selectivity factor, species A elutes faster than species B.
The selectivity factor is always greater than one.
● ii.Resolution
A characteristic of the separation of two adjacent
peaks.
Factor effecting resolution:
Temperature ramp rate
- Column length
- Carrier gas flow rate
- Film thickness
- Column internal diameter
- Ultra fast technology
iii.Peak shape
●Good peak shape can be defined as a symmetrical or
gaussian peak
●poor peak shape can include both peak fronting and

tailing.

• Good peak shape is important for....

Improved resolution (Rs)
More accurate quantitation
Longer usable column lifetime (based on system
suitability criteria)
 Factor effecting peak shape

● the initial column temperature,

● the column phase ratio, and
● the boiling points of the sample
● column selection
 iv.Operation Temperature

● Temperature programming
maintaining a low temperature for a short period of time, and increasing the
temperature to help force out the longer-‘sticking’ compounds.

● Isothermal
- use constant temperature throughout analysis
Operation Temperature
(a) low temperature (45 °C) - good
resolution initially - but too slow later

(b) higher temperature (145 °C) - much

faster, but poor resolution for early-
eluting species

In general - best results for temperatures

near boiling point of analyte.

If there is a wide range of boiling points

in the sample - then the best results are
obtained by temperature
programming as shown in (c), for the
same mixture, where the temperature
steps are as shown.
 current GC equipment
● GC-MS

● Several GC-MS have left earth. Two were brought to Mars by the Viking program.[14]
Venera 11 and 12 and Pioneer Venus analysed the atmosphere of Venus with GC-MS.[15]
The Huygens probe of the Cassini-Huygens mission landed one GC-MS on Saturn's
largest moon, Titan.[16] The material in the comet 67P/Churyumov-Gerasimenko will be
analysed by the Rosetta mission with a chiral GC-MS in 2014.[17]