GAS LIQUID CHROMATOGRAPHY

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

Column

gas system inlet column detector data system

Gas Carrier

Hydrogen

Air

Schematic Diagram of Gas Chromatography

Detector

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

Hydrogen Inlet

Column Effluent

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 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 by conduction to the molecules which contact with the filament.

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 separated compound elutes from the column , the thermal conductivity of the mixture of carrier gas and compound gas is lowered. The filament in the sample column becomes hotter than the control column. The imbalance between control and sample filament temeprature is measured by a simple gadget and a signal is recorded

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

Thermal Conductivity Detector

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

Electron Capture Detector

Analyses for pesticide, Insecticides, vinyl chloride, and fluorocarbons in foods.

Most sensitive detector (10-12 gram)

Electron Capture Detector

ECD detects positive ions of carrier gas by the anode electrode.
6 3

Ni emits particles.

Ionization : N2 (Carrier gas) + (e) = N2+ + 2e. The N2+ establish a base line X (F, Cl and Br) containing sample + (e) XIon recombination: X- + N2+ = X + N2, The base line due to the N2+ will decrease and this decrease constitutes the signal. The more the halogen containing X compounds in the sample, the less the N2+ in the detector

Electron Capture Detector

Electron Capture Detector

Chromatogram of Compounds from Fermented Cabbage

Chromatogram of Orange Juice Compounds

Gas Chromatography Application

Semi-Quantitative Analysis of Fatty Acids

C 18
10
Peak Area

C 16

8 6 4 2 0.5 1.0 1.5 2.0 2.5 3.0

Detector Response

C 14

Sample Concentration (mg/ml)

Retention Time
The content %of C fatty acids = 14 1 0 C + C+C 1 18 6
1 4

C4 1

= the content % of C fatty acids 14

Tentative Identification of Unknown Compounds

Mixture of known compounds

Response

1.6 min = RT Hexane

Octane

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

Advantages of Gas Chromatography

Very good separation Time (analysis is short) Small sample is needed - l 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 + CH3OH + H2SO4 O CH2 O C R CH O O C R Reflux

O R C O CH3 Volatile in Gas Chromatography

CH3ONa + CH3OH

O 3 R C O CH3 Volatile in Gas Chromatography

O CH2 O C R

Gas Chromatogram of Methyl Esters of Fatty Acids

Effects of OH groups of Carbohydrates

6 CH OH 2 5 4 HO 3 2 OH OH O 1 OH

Derivation of Glucose with Trimethylchlorosilane

6 CH2OH O 5 1 2 OH OH

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

4 OH HO 3

CH 3

Glucose

4 (CH3)3-Si-O

5HCl

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

Effects of Derivation

Time consumption Side reaction Loss of sample