Gas Chromatography (GC)

(Dr. Kuwat Triyana)

GAS LIQUID CHROMATOGRAPHY

Principles
Partition of molecules between gas
(mobile phase) and liquid
(stationary phase).

Most Common Stationary Phases

1. Separation of mixture of polar

compounds
Carbowax 20M (polyethylene glycol)
2. Separation of mixtures of non-polar
compounds
OV101 or SE-30 (polymer of
methylsilicone)

Gas Chromatography
Filters/Traps

Data system
H

RESET

Regulators

Syringe/Sampler
Inlets
Detectors

Gas Carrier

Hydrogen

Air

Column

gas
system
inlet
column
detector
data
system

Schematic Diagram of Gas Chromatograph

Schematic Diagram of Gas Chromatography

DETECTORS

Flame Ionization Detector (Nanogram - ng)

High temperature of hydrogen flame (H2

+O2 + N2) ionizes compounds eluted from
column into flame. The ions collected on
collector or electrode and were recorded on
recorder due to electric current.

chematic Diagram of Flame Ionization Detect

Exhaust

Chimney
Igniter

Hydrogen
Inlet

Collector Electrode
Polarizing Electrode
Column
Effluent

Schematic Diagram of Flame

Ionization Detector
Collector

Detector electronics

- 220 volts
Flame

Chassis ground

Jet
Column

Signal output

Thermal Conductivity Detector

Measures the changes of thermal
conductivity due to the sample (g).
Sample can be recovered.

Thermal Conductivity
Detector
Principal: The thermal balance of a heated
filament
Electrical power is converted to heat in a
resistant filament and the temperature will
climb until heat power loss form the
filament equals the electrical power input.
The filament may loose heat by radiation to
a cooler surface and by conduction to the
molecules coming into contact with it.

Thermal Conductivity Basics

When the carrier gas is contaminated by
sample
, the cooling effect of the
gas changes. The difference in cooling
is used to generate the detector signal.

Flow

Flow

The TCD is a nondestructive,

concentration sensing detector. A
heated filament is cooled by the flow of
carrier gas
.

Thermal Conductivity Detector

When a compound elutes, the thermal
conductivity of the gaseous mixture of
carrier gas and compound gas is
lowered, and the filament in the sample
column becomes hotter than the other
control column.
Its resistance increased, and this
imbalance between control and sample
filament resistances is measured by a
simple gadget and a signal is recorded

Thermal Conductivity Detector

Relative Thermal Conductivity

Compound

Carbon Tetrachloride
Benzene
Hexane
Argon
Methanol
Nitrogen
Helium
Hydrogen

Relative Thermal
Conductivity

0.05
0.11
0.12
0.12
0.13
0.17
1.00
1.28

Thermal Conductivity Detector

Responds to all compounds
Adequate sensitivity for many compounds
Good linear range of signal
Simple construction
Signal quite stable provided carrier gas glow
rate,
block temperature, and filament
power are controlled
Nondestructive detection

Electron Capture Detector

For pesticide analysis (picogram).

Accept electrons of carrier gas.

Electron Capture Detector

ECD detects ions in the exiting from the gas chromatographic
column by the anode electrode.
3

H or

63

Ni which emits particles.

Ionization : N2 (Nitrogen carrier gas) + (e) = N2+ + 2e

These N2+ establish a base line
X (F, Cl and Br) containing sample + (e) XIon recombination : X- + N2+ = X + N2
The base line will decrease and this decrease constitutes the
signal.
Insecticides, pesticides, vinyl chloride, and fluorocarbons

Electron Capture Detector

Electron Capture Detector

Gas Chromatography
Application

SEMI- QUANTITATIVE ANALYSIS OF FATTY ACIDS

Detector Response

C18

Peak Area (cm2 )

10

C 16

8
6

C14

4
2
0.5

1.0

1.5

2.0

2.5

Sample Concentration (mg/ml)

Retention Time
The content % of C14 fatty acids =

C
C + C + C

= the content % of C14 fatty acids

3.0

TENTATIVE IDENTIFICATION OF UNKNOWN COMPOUNDS

Response
Mixture of known compounds

Octane
1.6 min = RT
Hexane

GC Retention Time on Carbowax-20 (min)

Response

Unknown compound may be Hexane

1.6 min = RT

Retention Time on Carbowax-20 (min)

Decane

Retention Times
Response

RT= 4.0 min on SE-30

Hexane

GC Retention Time on SE-30

Response

RT= 4 min on SE-30

Unknown compound

GC Retention Time on SE-30

GLC ADVANTAGES
1. Very good separation
2. Time (analysis is short)
3. Small sample is needed - l
4. Good detection system
5. Quantitatively analyzed

DISADVANTAGES OF GAS CHROMATOGRAPHY

Material has to be volatilized at 250C without decomposition .

Methylester

Fatty Acids

R C OH + CH3 OH + H2 SO4

Reflux

R C O CH3
Volatile in Gas
Chromatography

O
CH2 O C R
O
CH

O C R
O

CH2

O C R

CH3 ONa
+

CH3 OH

O
3 R C O CH3
Volatile in Gas
Chromatography

Gas Chromatogram of Methyl Esters of Fatty Acids

The Effects of OH groups of

Carbohydrates
6
CH2OH
O
5

4
OH
HO
3

6
CH2OH
O
5

2 OH 4
OH
OH
HO
3

1
2 OH
OH

Derivation of Glucose with Trimethylchlorosilane

6
CH2OH
O
5

4
OH
HO
3

CH
1 +

Si

5Cl

2 OH
OH

CH

Glucose

4
(CH3)3-Si-O

3
CH

Trimethylchlorosilane
6
CH2O-Si(CH3)3
O
5
O-Si(CH ) 1
3 3

2 O-Si(CH3)3
O-Si(CH3)3

5HCl

Effects of Derivation
1.Time
consumption
2.Side reaction
3.Loss of sample

THIN LAYER CHROMATOGRAPHY

Stationary Phase ---------> Silica Gel
Mobile Phase -------------> Solvent
(developing)

THIN LAYER CHROMATOGRAPHY

DEVELOPING
CHAMBER
SPOT

SOLVENT

Solvent
Front

1.1 cm
5.5 cm

Origin

Rf =

Distance from starting origin to center of zone

Distance from starting origin to solvent front
5.5
11

= 0.5

Thermal Conductivity Detector

The detector contains two filaments: one
exposed only to carrier gas, while the other
is exposed to the carrier gas for sample
analysis.
When the gas for the sample analysis is
only carrier gas , the two filaments can be
balanced.
Instead of a direct measurement of
filament temperature, the filament

Thermal Conductivity Detector

The ability of a colliding molecule to
carry off heat depending on its thermal
conductivity. Hydrogen and helium have
high thermal conductivity and therefore
will be more efficient at cooling a
heated filament than other gases will

Thermal Conductivity Detector

The TCD will respond to any substance

different from the carrier gas as long a
concentration is sufficiently high enoug

Thermal Conductivity Detector

Thermal Conductivity Detector

Electron Capture Detector

Electron capture compound, X (highly electonegative element), tends to
capture free electrons and increase the amount to ion recombination
X (F, Cl and Br) + e XIon recombination : X- + N2+ = X + N2
The current will decrease and this decrease constitutes the signal.

Halogens, lead, phosphorous, nitro groups, silicone and polynuclear

aromatics.
Insecticides, pesticides, vinyl chloride, and fluorocarbons

Electron Capture Detector