Gas Chromatography

Yao Zhongping
Office: Y832
Email: bczpyao@[Link]
 Outline
• Overview of gas chromatograph
• Carrier gas
• Sample introduction system
• Column
• Detector

• Applications
• Summary

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After this session, you are expected to be
able to

• Describe basic components of a gas chromatograph and

the working principle and role of each component

• Apply gas chromatographic techniques to solve problems

3
Reference book:

Skoog, D. A.; Holler, F. J.; Crouch, S. R. Principles of

Instrumental Analysis, Thomson Brooks/Cole, 6th edition
2007.

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Gas chromatography
• Applicable to “volatile” compounds only.
Analyte should have appreciable vapour pressure and
stability at “injector” temperature.

• The retention of sample component depends on:

▪ Volatility
▪ The degree of interaction with stationary phase
Hydrogen bonding
Dipole-dipole
Dipole-induced dipole
Induced dipole-induced dipole
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Gas Chromatograph

Gas (Mobile phase) Supply System

Injection System
Heated,
Chromatographic Column & Column Oven Temperature
Controlled
Detector(s)

Signal Conditioning System

To convert signals for processing by the data system.
Data System
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6
 Electronics
(for signal
conditioning)

Schematic of a gas chromatograph

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Computerized Recorder Gas Chromatograph
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 A commercial gas chromatographic system

Oven temperature
control module

Gas flow
control
module

Column
Oven Chromatographic
column
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 Injectors
Detectors

Control &
Signal
Conditioning

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Carrier gas (MOBILE PHASE)

• Should be inert and do not interact with analytes.

Usually helium or nitrogen, sometimes hydrogen and argon
• Should be pure.
Free from water vapour, oxygen and impurities
• Commonly supplied by a compressed gas cylinder (initial
pressure ~150 bar) or gas generator.

• Constant flow rate / head pressure

Controlled by pressure / flow controller

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Injector (sample introduction device)

• Fast vaporization of sample

• Effective transfer of sample to column

• Non-discriminating
The same effects on the components with different boiling points.

Analyte should have appreciable vapour pressure at

injector temperature without decomposition.
Analysis applicable to “volatile” compounds only.

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 Heater

Simple on-column injector for packed column

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 Capillary column
Fins for heat
injector
dissipation

Packed column
injector

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A Simple Septum Injector
Rubber
Septum

To column

Hole for
syringe
insertion
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Sample Injection Syringe
Volume: usually 0.5 – 10 µL

Cheap
Easily handled
Repeatable
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Problems associated with sample injection into
capillary column

• Column can accept only a very small amount of sample.

• Broadening of sample peak due to sample overloading.

• Very sensitive to contaminants.

Solution

• Inlet splitting

• Purging
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Split injector for capillary column

• Bleeding from
septum is “blown
away” by a purge
gas stream.

• Part of the sample

vapour is allowed
to escape through
the split outlet at
sample injection.

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Column
Cylindrical tubing containing the stationary phase

Material for making the column body

• Inert
• Can withstand operating temperature
• Common material: Stainless steel, glass, silica (quartz)
Other used material: Copper, aluminum, Telfon (PTFE)

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Column

packed column capillary column

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Packed Column
The chromatographic column is firmly and uniformly packed with
coated or uncoated solid particles—stationary phase that are
responsible for chromatographic separation.

Solid stationary phase or Liquid stationary phase

solid support particles (coated or bonded)

Length : 1 to 10 meters Diameter: usually 2 – 6 mm

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Open Tubular (Capillary) Column
Wall-Coated Open Tubular (WCOT) Column

Column tubing (usually

Liquid stationary phase
made from quartz)
coated/bonded to wall

Polyimide outer
coating (for strength)

Diameter: 0.1 – 0.53 mm i.d.

Length: 5 – 100 meters
Stationary phase thickness: 0.1 – 5 µm
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Comparison between packed and capillary (WCOT) columns
Type of Column
WCOT Packed
Length, m 5 – 100 1–6
Inside diameter, mm 0.1 – 0.53 2–6
Efficiency, plates/m 2000 – 4000 500 – 1000
Total number of plates (20 – 400) × 104 (1 – 10) × 103
Sample size, ng 10 – 75 10 – 106
Relative back pressure Low High
Relative speed Fast Slow
Flexible? Yes To some extent
(steel)
Capillary column requires less sample and is faster and more efficient
for separation.
Packed column is not commonly used now. 23
Liquid Stationary Phase
Non-volatile (very low vapour pressure) and inert liquids.
even at 200 – 300°C

• The liquid should be thermally stable.

• The liquid should have good solvent characteristics for the
analytes.
• Usually polymeric.
Oils, greases or even waxes at room temperature
• Liquid stationary phases are applied as thin layer coated onto
the support.
• There are a wide variety of choices of different polarity.
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Examples of commonly used stationary phase
Polysiloxane (OV phases)

OV-1 R = methyl (non-polar)

OV-5 R = 5% phenyl, 95% methyl
OV-17 R = 50% methyl, 50% phenyl (semi-polar)

Polyethylene glycol (Polar stationary phase)

Carbowax 20M (M. W. ~20,000) 25

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 26
Polarity increases
Choosing Liquid Stationary Phase
Major Factors:

• Maximum operating temperature

• Differential solubility

Like dissolves like

Compounds dissolve better in the stationary liquid phase

will be more retained.

alkanes, olefins, aromatics, esters, alcohols, acids

polarity

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 A (Methanol) Polar, b.p. 65°C
B (Ether) non-polar, b. p. 36°C
C (Methyl acetate) Semi-polar, b.p. 57°C

A BC

1. OV-1
(non-polar)

Time

2. CARBOWAX
(polar)

Time
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Bonded Phase
Chemically bonded stationary phase
R
Si OH Si O Si R
Cl R R
O + 2 -2HCl O
Si R
Si OH R R Si O Si R
R

Silanization

Fused Silica Bonded Phase Column

Polyimide Coating
Fused Silica Capillary Tube 0.1 - 0.5 mm I.D.

Chemically Bonded Stationary Phase Layer~

1 µm thick

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Other Capillary Columns
Support-coated open tubular (SCOT) column

Glass/Silica Capillary Tube

Stationary phase-coated inert

solid particles adhering to
the column wall

Porous layer open tubular (PLOT) column

Glass/Silica Capillary Tube

Etched Porous Layer

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Detector

Detector

Concentration or mass rate

dependent

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Mass Rate Dependent detector
• Responds to the mass of solute passing through it per unity
time.

• Detector response (peak area) is unaffected by change in

flow rate of the mobile phase.

e.g. Flame Ionization Detector (FID)

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Concentration Dependent Detector

• Responds to the mass of the solute per unit volume of

mobile phase passing through it.

• Detector response (peak area) is affected by change in

flow rate of the mobile phase.

e.g. Thermal Conductivity Detector (TCD)

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Characteristics of an ideal detector

• Adequate sensitivity

• Wide linear and dynamic range

• Low noise

• Short response time

• Good stability and reproducibility

• Being nondestructive to samples

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Noise
• Unwanted signal fluctuation.
• Defined as standard deviation of the signal.
• Sources:
▪ Fluctuation in power supply
▪ Random arrival of photons, etc

• Signal-to-noise ratio (S/N)

The ratio of the average value of the output signal
to its standard deviation.

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Signal to Noise Ratio

sample
noise

Average signal
Signal

blank

Time

average signal
S/N =
noise
Limit of detection (LOD)
The lowest concentration / amount of analyte that can be
detected (typically gives a signal with S/N = 3)
Signal

Time

Limit of quantification (LOQ)

The lowest concentration / amount of analyte that can be
quantified (typically gives a signal with S/N = 10)
Sensitivity

An instrument/method is more sensitive, if it is able to measure

smaller changes in the content/concentration of the analyte

Sensitivity may also be defined by the slope of the calibration graph

Response

Concentration
Dynamic range & linear range

Linear Range
Signal

Concentration
Linear range: concentration range that signal has a linear relationship
with concentration.
Largest concentration in the linear range
Dynamic range =
Smallest concentration in the linear range

e.g., linear range: 10-5 – 10-2 M, Dynamic range: 103 (3 orders of

magnitude)
Thermal Conductivity Detector (TCD)

• Detection based on the difference in thermal conductivity

between the analyte and the mobile phase (carrier gas).

• The resistance/voltage across an electrically heated

filament is measured.
• It is almost a universal detector.

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 Small molecule
high thermal
Heat transfer conductivity
by conduction
Carrier flow

BLOCK
Large molecule
Heat transfer low thermal
by conduction conductivity

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 Block

When an analyte elutes and

the thermal conductivity of
the effluent is reduced.

The filament heats up and

changes resistance, and a
signal is thus observed.

Filament

Thermal Conductivity Cell 42

Carrier Gas for TCD

• Carrier gas should have either very HIGH or very

LOW thermal conductivity.

Large difference in thermal conductivity between

analyte and carrier  HIGH SENSITIVITY

• Common carrier gas: hydrogen /helium.

Thermal conductivity 6-10 times that of organic compounds

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Features of TCD
• Simple construction.

• Nondestructive detection.

• Wide dynamic range (~105).

• Relatively low sensitivity (~10-8 g/mL).

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Ionization Detector
Electrodes
Ionization
Source
+
- + + - + -- -
+
- - + -+ - - -
+ - +
Carrier Gas Flow + +
-
- - + - + - - +
+
+
+ - -+ +
- + - + - + +
-
• Analyte is ionized by some means.
• The ion current flowing across 2 electrodes (kept at a
high potential difference) is measured. 45
Schematic of Flame Ionization Detector (FID)
Resulting current:
Ionization is ~10-12 A
induced by
burning in a
Electrometer
hydrogen /
(+)
air flame

Resistor
Flame
Igniter -
( )

Air

Hydrogen Carrier
47
Non-combustible compounds: no signal

NH3 He CO
H2O Flame
Ionization No
CS2 O2 Air Response
Detector
H2S N2
SO2 NH3

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Features of FID
• High sensitivity (~10-13 g/s).
• Very wide linear response range (~107).
• Responds to the number of carbon atoms entering the detector
per unit time. (mass rate dependent)
▪ Functional groups, such as carbonyl, alcohol, halogen, and
amine, produce fewer / no ions.
▪ No signal from non-combustible gases.
Such as H2O, CO2, SO2, noble gases, and NOx.
• Applicable for most organic samples.
Including those contaminated with water and the oxides of nitrogen
and sulfur.
• Most widely used detector for GC.
• Destructive detector and requirement of additional gases. 49
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 e-
- e- +

CATHODE e- e- ANODE
e-

   


Radioactive
β-emitter

Ionization is induced by β-particles emitted from a radio-isotope

Principle of ECD
• The sample elute from a column is
passed over a radioactive β emitter, + electrode
usually nickel 63.
• β particles from the emitter cause
ionization of carrier gas and the
- electrode
production of a burst of electrons. β –emitter
(63Ni )
Carrier gas: Ar-CH4, He-CH4, N2

CH4 CH4+ + e

• In the absence of organic species, a constant standing current

between electrodes results from this ionization process.
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In the presence of sample molecule (A-X)

A—X + e A—X-

The standing current is reduced by the decrease of the free electrons.

Standing current

Sample molecules with highly electronegative groups/atoms

can be detected by this detector.
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Features of ECD

• Highly sensitive to electronegative functional groups.

e.g. halogens, peroxides, quinones, and nitro group.

• Non-destructive.

• Radioactive hazard.
Using ß-emitting radioactive substances e.g. 63Ni, 3H.

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Mass spectrometry detector (MSD)

• Each component is subsequently ionized in ion source.

Electron Ionization (EI) or Chemical Ionization (CI) ion source
• The generated ions are separated by mass analyzer and
detected by detector, and a mass spectrum is obtained for
each component. 54
Features of mass spectrometry detector
• Database containing over 200,000 standard EI spectra
allows spectral comparison and rapid identification of
components by database library.
e.g. NIST library
• Specific mass spectrometry techniques allow rapid, selective
and sensitive quantitation of components.
e.g. selected ion monitoring (SIM)
• One of the most powerful detector for GC.

• High cost.

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Qualitative analysis
• To establish the purity of organic compounds.
Additional peaks Impurities
• To identify components by retention times.
Problems: Reproducibility of retention time
Similar retention times of different compounds
Usually used for confirming presence or absence of a suspected
compound in a mixture. (by co-injection with the standard)
• To identify components by coupling GC with detectors such as
mass spectrometry.
GC-MS: Premier technique for analysis of mixture.
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Quantitative analysis
• Based on peak height
More easily measured. Related to peak widths
• Based on peak area
Independent of peak broadening. Preferred method.

Quantitative methods
• The external-standard method
• The internal-standard method
• The standard addition method
• The area-normalization method
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External standard method

• Construct a calibration curve using the

standard sample of different
concentrations.

• Determine the concentration of the sample

to be quantified from the calibration curve
using the measured intensity of the
sample.

The standard sample and the sample to

be quantified are measured separately
under the same conditions.

• Variations in sample treatment and

instrumental analysis can cause errors. De Hoffman, E. and Stroobant, V. Mass Spectrometry: Principles
and Applications, John Wiley & Sons, 3rd ed., 2007, p266.

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Internal standard method
Similar procedures to the external method,
but in each measurement, an internal
standard is added to correct the variations.
• A calibration curve is obtained by
measuring samples containing the fixed
quantity of the internal standard and
increasing quantities of the external
standard (usually the same as the
compound to be quantified).
• The sample to be quantified is then
measured, also with a constant quantity
of internal standard added to it.
• Intensity ratio with the internal standard
De Hoffman, E. and Stroobant, V. Mass
is used for the quantitation, to Spectrometry: Principles and Applications, John
compensate the variations in sample Wiley & Sons, 3rd ed., 2007, p267.

treatment and instrumental analysis.

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Standard addition method
• Add different concentrations (e.g., 0, s1, s2, s3, s4) of the standard (the same
as the compound to be quantified) to each sample of the same volume.
• Measure the intensity of each solution and plot intensity against the added
concentration of the standard.
I
x x x x x
+ + + + +
0 s1 s2 s3 s4

x+0 x+s1 x+s2 x+s3 x+s4 s

The x-intercept of the extrapolated line is the concentration of the sample

I = a(x+s) When I = 0, x = -s.
• Usually used if the matrix is present for the sample and not for the
standards, e.g., urine samples, soil samples.
• Normally is not as sensitive as the standard method since it relies on the
intensity change to determine the sample concentration. 60
The area-normalization method

Area A
= molar response (Rm), area per unit mole
Mole A

(Area B)/RmB
× 100%
Mole% of B =
(Area A)/RmA + (Area B)/RmB + (Area C)/RmC

Sample component area Rm relative mole mole%

(× 100)
Benzene 35 100 35.0 41.07
Heptane 58 143 40.6 47.65
Methyl hexane 13 136 9.6 11.26

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 Temperature Programming

Temperature 2
Column Temperature

Isothermal stage 2

Temperature 1
Isothermal stage 1

Time
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Temperature programming
45 oC

120 oC

180 oC

30 oC

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Advantages of temperature programming

• Analysis of wide boiling range samples

• Reasonable analysis time

• Better resolution of peaks

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 Gasoline

Peak Identification
1. Benzene
2. Toluene
3. Ethylbenzene
4. m + p-Xylene
5. o-Xylene

Stainless Steel, 6ft × 1/8" OD, 10% TCEP on 100/120

Column:
Chromosorb P-AW
Temp: 50°C (3 min hold) to 130°C at 5°C/min
Carrier: Nitrogen at 29 mL/min
Detector: FID
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Source: Alltech Chromaccess database
 Peak Identification
Unleaded Gasoline 1. n-Butane
2. Isobutane
3. Pentane
4. 2-Methylpentane
5. 3-Methylpentane
6. Hexane
7. Methyl cyclopentane
8. 2,4-Dimethylpentane
9. Benzene
10. 2-Methylhexane
11. 3-Methylhexane
12. n-Heptane
13. Toluene
14. Ethylbenzene
15. m-Xylene
16. p-Xylene
17. 2-Methyloctane
18. o-Xylene
19. n-Nonane
Column: Heliflex® AT-1, 30 m × 0.25 mm × 1.00 um
20. Propylbenzene
Temp: 35°C (12 min hold) to 200°C (20 min hold) 21. 1,2,4-Trimethylbenzene
22. n-Decane
at 4°C/min
23. n-Undecane
Carrier: Helium at 0.65 mL/min (22 cm/sec) 24. 2-Methylnaphthalene
25. 1-Methylnaphthalene
Detector: FID at 275°C 26. Dimethylnapthalenes

Source: Alltech Chromaccess database 66

[Link]
 Organophosphorus Pesticides on AT-5ms
(Composite Mix) Peak Identification
1. TEPP
2. Sulfotepp
3. Monocrotophos
4. Dimethoate
5. Dioxathion
6. Terbufos
7. Phosphamidon
8. Malathion
9. Parathion
10. cis-Chlorfenvinphos
11. trans-Chlorfenvinphos
12. Ethion
Heliflex® AT-5ms, 30 m × 0.25 mm × 0.25 um
Column: 13. Famphur
Part No. 15807
14. Carbophenothion
Temp: 120°C (3 min hold) to 285°C (4 min hold) at 15. Phosmet
5°C/min
16. EPN
Carrier: Helium at 0.8 mL/min (26 cm/sec) 17. Leptophos
Detector: MSD at 300°C 18. Azinphos-ethyl

Source: Alltech Chromaccess database 67

[Link]
Summary
• GC is applicable for
volatile and thermally stable compounds
• Components of a mixture are separated based on their
volatility and interactions with the stationary phase
• A gas chromatograph consists of
Gas supply system Injection system Column and oven
Detector Signal conditioning system Data system
• GC can be used for
separation, qualitative and quantitative analysis
• Performance indicators of detectors:
Noise, S/N, LOD, LOQ, sensitivity, dynamic range, linear range
• Quantitative methods:
External-standard method Internal-standard method
Standard addition method Area-normalization method 68