GAS

CHROMATOGRAPHY
MMS 3201
AQUATIC METHODOLOGY AND
INSTRUMENTATION
Izwandy Idris, Blue Cabin,
ext. 3467, izwandy.idris@umt.edu.my

Introduction
What is Chromatography?

Separation of compounds (solutes) into separate

components for identification (qualitative) and
concentration determination (quantitative)

Several techniques including such as Thin Layer,

Size-exclusion, HPL and Gas Chromatographies.

Gas chromatography separating and analysing

volatile compounds that can be vaporized without
decomposition

GAS CHROMATOGRAPHY

Suitable for volatile and thermally stable (<450 oC)

compound

Application in aquatic science: detection of

Disinfection by-Products, taste odour components,
insecticides, pesticides, hydrocarbon, fatty acids etc.

Sample of organic compounds is injected into an inert

gaseous mobile phase (known as the carrier gas) for
analysis.

The sample is carried through a packed or capillary

column and the components in the sample are
separated based on their ability to distribute between
the stationary phase and the mobile phase.

GC analytical technique
2.

Separation processes
i. Different adsorption with column packing
materials.
ii. Different Boiling point (Temp programming
System).
iii. Different in weight (Molecular weight)

Sample injected
(A, B &C)

B
C
A
Carrier (inert) gas Initial column temp set at 60oC
The boiling points for
A=70oC

B=100oC & C= 150oC.

If we increase the column temperature

from 60oC to 200oC at the rate of
10oC/min, the results of the
chromatogram will be
Retention Time, RT
A
B
C

Peak
height

Time, min.

INJECTION PORT

GC SEPARATION

Common inert mobile phase Neon (Ne), Argon (Ar), Helium

(He) and Nitrogen gas (N2)

Inert gas no chemical reaction with mobile phase

(compound) and stationary phase.

Chromatography column two types i.e. packed and

capillary

Types of Columns for GC

Capillary Columns

A.k.a open tubular column - made from

fused silica coated with a protective
polymer.

Up to 100 meter, diameter 50 300 m.

TYPES OF GC COLUMN

(0.01 0.053cm)

(0.3-0.4cm)

Types of Columns for GC

Packed Column

Made from glass, stainless steel, copper or

aluminum, typically 2 6 m long, internal
diameter 2 4 mm.

Filled up with particulate solid with diameter

ranging between 30-350 m. Smaller solid
particle, > separation

TYPES OF GC COLUMN

(0.01 0.053cm)

(0.3-0.4cm)

Comparison Of column

Capillary column

Packed column

Capillary Column (Shimadzu 2010)

The Stationary Phase

Use high molecular weight, thermally

polymer, and chemically inert

stable

Coating the inner tube (capillary) or solid support

material

Solutes with different of boiling points are easily

separated. Two solutes with the same boiling points
could be separated if the stationary phase reacts
with one of the solutes

Optimal detection similar polarity with solute

(increase separation and absorption)

Non-polar solutes easier separation with nonpolar stationary phase

Injection of Sample
1. Sample must be volatile.
2. The analyte must be present at an
appropriate concentration.
3. Injected sample must not be degradable
in the column.

Temperature Control (Temp Programming)

Important for good separation

Column inside a thermo-stated oven

Isothermal separation column is

maintained at a constant temperature

Multi separation temp is set slightly below

the lowest boiling solute and increase
gradually (Temperature programming)

DETECTORS FOR GC
1.

Thermal Conductivity Detector

based on mobile phases thermal conductivity
Helium is selected mobile phase high thermal conductivity
When a solute is eluted from the column, the thermal
conductivity of the mobile phase decreases change in
conductivity
TCD gives a signal for any solute which has different thermal
conductivity from helium
Can be used to detect any compound

DETECTORS FOR GC
2.

Flame Ionization Detector (FID)

Combustion of organic compounds in H2/air flame - flame
rich in electrons and ions.
Ions are detected with electrically charge metal detactor (DC:
~300 v), more current across detector high rate of
ionisation high concentration
used only for organic compounds

Sensitivity is 2 3 lower than the TCD.

DETECTORS FOR GC
3. Electron Capture Detector (ECD)
consists of a beta emitter (electron) such as 63Ni.
Emitting electrons ionize the mobile phase (usually
N2), production of additional electrons electric
current

When the eluted solute pass through the ECD, the

electric current in the ECD decrease and a signal
is obtained
Selective detector for solutes with electronegative
functional groups (halogenated compounds and
nitrogen groups) pesticides (environment),
pharmaceutical.

OM

OM

/Hydrocarbons

H2 + O2

N2

Example of Chromatogram-Petroleum
Hydrocarbon Analysis
Petroleum hydrocarbons are extracted from sea
water and fractionated into
1. Aliphatic hydrocarbon &
2. Aromatic hydrocarbon fractions using
Column Chromatography.
GC-FID with capillary column (30 m) and
temperature programming 20 min (70 oC),
increase to 300oC with a rate of 8oC/min and hold
at 300 oC for 30 min

Aliphatic Hydrocarbons
FID2 B, (JOHNSON\SIG11155.D)
pA

C27

C18

150

C29

C19

C28

C23

C24

C26

C21

200

C22

250

10

20

C32

C20

C16

C10

C9

C8

50

ISTDi

C17

100

30

40

50

60

70

min

Polyaromatic Hydrocarbons
FID2 B, (JOHNSON\SIG11026.D)
pA
120

BkF

100

Pyr

80

60

ISTDI

ISTDII

20

10

20

30

40

BbF
IP

ISTDIII
BaA

40

50

60

70

min

Qualitative & Quantitative Analysis

1.
2.

3.
4.

Use Reference Standard Material such as

EPA 16 Aromatic Hydrocarbons
RSM is analyzed with the GC under the
same condition set for the analysis.
RT value and the intensity of peak are
recorded
Compare with the samples peak

Retention Value(RT) 11.9 minutes

Aliphatic Hydrocarbons External Standard

C32

C29

C28

C27

C26

C24

C23

C22
C21
C19

C8

300

C18

C16

400

C17

C13

500

C14

C12

C30

C20

C25

C9

600

C11

pA

C15

C10

FID2 B, (JOHNSON\SIG11151.D)

200

100

10

20

30

40

50

Polyaromatic Hydrocarbons External Standard

pA

Ace
Acy

Na

FID2 B, (JOHNSON\SIG11032.D)

450

400

350

300

250

Flu

200

10

20

30

40

BbF

dBahA
BghiP

50

BkF
BaP
IP

Pyr

100

BaA Chyrs

AnPh

FI

150

50

GC-MS (Identification of Organic Compounds)

1.

Sample with unknown solutes

2.

Effluent from GC is introduced into the mass

spectrometers ionization chamber mobile phase
removal

3.

Ionization chamber ionization of molecules and

separated by mass to charge ratio

4.

Selective monitoring detector is used to detect the

specific mass-to-charge ratio produced electricity
current

GC MS(Shimadzu 2010)

GC-MS FLOW DIAGRAM

GC MS 3D VIEW

Gas Chromatograph Mass Spectrometer

Total Organic Carbon Analyzer

(TOC)

Analysis of organic substances in

- Water
- solid including tissues and sediment
IR light absorption

Organic + oO2
600 C

CO2

Infrared Analysis

TOC

Video on Gas Chromatography

https://www.youtube.com/watch?v=08YWhLTjlfo