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 Carrier Hydrogen Air

Column

gas system inlet 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 Inlet 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 (mg). 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, concentration sensing detector. A heated filament is cooled by the flow of carrier gas . 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

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 Benzene Hexane Argon Methanol Nitrogen Helium Hydrogen

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.
3H

or 63Ni 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 3.0

Sample Concentration (mg/ml)

Retention Time
C C + C C +

The content % of C fatty acids = 14

= the content % of C fatty acids 14

TENTATIVE IDENTIFICATION OF UNKNOWN COMPOUNDS

Response Mixture of known compounds

Octane 1.6 min = RT Hexane

Decane

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

DISADVANTAGES OF GAS CHROMATOGRAPHY

Material has to be volatilized at 250C without decomposition.
Fatty Acids Methylester

O R C OH + CHOH + H SO4 3 2 R Reflux

O C O CH 3 Volatile in Gas Chromatography

O CH 2 O C O CH O C O CH 2 O C R R + CH 3 OH R CH3 ONa 3R O C O CH 3

Volatile in Gas Chromatography

Gas Chromatogram of Methyl Esters of Fatty Acids

The Effects of OH groups of Carbohydrates

6 CH2 OH O 5 4 OH HO 3 1

2 OH 4 OH OH HO 3

6 CH2 OH O 5 1

2 OH OH

Derivation of Glucose with Trimethylchlorosilane

6 CH2 OH 5 4 HO OH 3

O
1 2 OH OH

CH 3 + 5Cl Si CH 3 Trimethylchlorosilane
6 CH2 O-Si(CH3)3 O 5 O-Si(CH ) 1
3 3

CH 3

Glucose

4 (CH3)3-Si-O

5HCl

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

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

THIN LAYER CHROMATOGRAPHY

Stationary Phase --------->

Silica Gel

Mobile Phase -------------> (developing)

Solvent

THIN LAYER CHROMATOGRAPHY

DEVELOPING CHAMBER

SPOT

SOLVENT

Solvent Front

1.1 cm 5.5 cm

Origin

Distance from starting origin to center of zone Rf = 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 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