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)

3. Methylester of fatty acids

DEGS (diethylene glycol succinate)
 Gas Chromatography
Filters/Traps Data system
H

RESET

Regulators Syringe/Sampler

Inlets

Detectors  gas system

Gas Carrier
Hydrogen

 inlet
Air

Column  column
 detector
 data system
Schematic Diagram of Gas Chromatography
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.
Schematic Diagram of Flame Ionization Detector

Exhaust

Chimney

Igniter Collector Electrode

Polarizing Electrode

Hydrogen Column
Inlet 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 (µ 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

The TCD is a nondestructive, When the carrier gas is contaminated by

concentration sensing detector. A sample , the cooling effect of the
heated filament is cooled by the flow of gas changes. The difference in cooling
carrier gas . is used to generate the detector signal.
Flow

Flow
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 Relative Thermal
Conductivity
Carbon Tetrachloride 0.05
Benzene 0.11
Hexane 0.12
Argon 0.12
Methanol 0.13
Nitrogen 0.17
Helium 1.00
Hydrogen 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 β particles.
Ionization : N2 (Nitrogen carrier gas) + β (e) = N2+ + 2e
These N2+ establish a “base line”

X (F, Cl and Br) containing sample + β (e)  X-

Ion 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

D e te c to r R e s p o n s e
C1 8 (2 )
P eak A reacm
10

C1 6 8

C1 4 4

0.5 1.0 1.5 2.0 2.5 3.0

S am ple C on centratio n (m g/m l)
R e te n tio n T im e

C1
4
The content %of C14fatty acids = ∗1
0
C + C+
1
61
C 8
1
4

= the content % of C fatty

14 acids
TENTATIVE IDENTIFICATION OF UNKNOWN COMPOUNDS

Response

Mixture of known compounds

Octane
Decane
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)

Retention Times
R e spo n se

R T = 4 .0 m in o n S E -3 0
H ex an e

G C R ete n tio n T im e o n S E -3 0

R e spo n se

R T = 4 m in o n S E -3 0

U n k n o w n c o m p o u nd

G C R e ten tio n T im e o n S E -3 0
 GLC ADVANTAGES

1. Very good separation

2. Time (analysis is short)
3. Small sample is needed - µ l
4. Good detection system
5. Quantitatively analyzed
DISADVANTAGES OF GAS CHROMATOGRAPHY

Material has to be volatilized at 250C without decomposition.

Fatty Acids Methylester

O O
R C OH +
CH 3 OH +H 2 SO 4 R C O CH 3
Reflux Volatile in Gas
Chromatography
O
CH 2 O C R

O CH 3 ONa O
CH O C R + CH 3 OH 3 R C O CH 3

Volatile in Gas
O Chromatography
CH 2 O C R
Gas Chromatogram of Methyl Esters of Fatty Acids
The Effects of OH groups of Carbohydrates

6
CH2OH
O
5 6
4 CH2OH
OH 1
HO O
OH 5
3 2 4
OH OH 1
HO
OH
3 2
OH
Derivation of Glucose with Trimethylchlorosilane

6
CH2OH
O CH 3
5
4
OH 1 + 5Cl Si CH 3
HO
OH CH 3
3 2
OH
Glucose Trimethylchlorosilane
6
CH2O-Si(CH3)3
O
5
4 + 5HCl
O-Si(CH ) 1
3 3
(CH3)3-Si-O
3 2 O-Si(CH3)3
O-Si(CH3)3
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

1.1cm

5.5cm

Origin

Distancefromstartingorigintocenter of zone
Rf =
Distancefromstartingorigintosolvent front

5.5
= =0.5
11
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 resistant, which is a function
of temperature, is measured.
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 as its
concentration is sufficiently high enough.
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  X-

Ion 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