What is Gas Chromatography?

Gas chromatography or gas-liquid chromatography is a chromatographic technique that used for the separation of volatile organic compound. It involves separation, identification and quantification of the chemical compound. The separation of the mixture is done due to the difference in the interaction between the analyte with stationary phase as it carries by the mobile phase. The chemical compounds or analyte where in vapour state is distributed between the stationary phase and the mobile phase. The mobile phase does not interact with molecule of the analyte; its only function is to transport the analyte through the column. In this instrumentation of chromatography, the mobile phase is gas while the stationary phase is liquid that held inside the column. There were few components that made up gas chromatography which is carrier gas(mobile phase), injection system, oven(where stationary phase located), column, detector and signal processing and read out.

Components of gas-liquid chromatography

(from http://teaching.shu.ac.uk)

1. Mobile phase: Mobile phase must not react with the analyte. Its function is to carry analyte through the stationary phase or the column. It also refers as carrier gas.

Carrier gas It must be chemically inert, for example helium, nitrogen, and hydrogen. Pressure regulators, gauges and flow meter were associate with the gas supply. It usually contain molecular sieve to remove water or other impurities.

2. Sample injection system. It is where the sample is being introduced to the system. Column efficiency requires that the sample to be of suitable size and introduced as a plug of vapor. A Slow injection of oversized samples will causes results to have a band spreading and poor resolution. Commonly used method for injection of sample is by the aid of microsyringe to inject a liquid or gaseous sample through a self-sealing, silicone-rubber diaphragm or septum into a flash vaporizer port located at the head of the column where the sample port is ordinarily about 50oC above the boiling point of the least volatile component of the sample. Thus before running the sample, it is crucial to know the boiling point of the analyzed sample. There are few types of injection port that is used which is flash vaporization, splitless capillary injector that is commonly used, direct injector that used in trace analysis, cool on-column injector, autosampler and head space. 3. Oven In gas chromatography, optimum temperature depends on the boiling points of the sample components. Thus the control of the temperature depends on the type of compound analyzed which is its boiling point. A temperature that is roughly higher than the average boiling point of the sample results in a reasonable elution period. Temperature programming is used to reduce the retention time of the sample in the column and overcome seperations problem.

4. Column It is where the stationary phase of the system located. Column is the location where separation of the analyte occurs. There are two types of column in gas chromatography which is packed column and capillary column. Packed column: It is less commonly used and made of glass and steel. It length range from 1 to 5 meter with internal diameter if 2 to 4 mm. These column is densely packed with uniform, finely divided solid support, coated with thin layer (0.05 to1m) of stationary liquid phase. It also accommodate larger samples. Inside the packed column, the carrier gas flow between 10 40 mL/min. However, it is not suitable for trace analysis. It also contain an inert and stable porous support on which the stationary phase can be impregnated or bound. Capillary column Unlike packed column, capillary column is widely used in gas

chromatography analysis. It also known as open tubular calumn. Its length is between 10 and up to 100 meter. They were coiled around a light weight of metallic support. There were a few types of capillary column which is: FSOT (Fused Silica Wall Coated). Its internal diameter range from 0.1 to 0.3 mm WCOT (Wall Coated) Internal diameter range from 0.25 to 0.75 mm SCOT (Support Coated) Its internal diameter is 0.5 mm PLOT

5. Detector It measure the eluents after as they evolve from the column. It is where the components of mixture or analyte is identified. There were few factors that made up a good detector. For ideal detector: a) it need high reliability & ease to use. b) Similarity response toward all solutes or alternatively a high predictable and selective response toward one or more classes of solute. c) Detector should also be nondestructive where there is no damage to column is occur. d) High sensitivity e) Have a good stability and reproducibility. f) Linear response to solutes that extends over several orders of magnitude. g) Temperature range from room temperature to atleast 4000C. There are three types of detectors that is used in gas chromatography: a) Thermal conductivity detector(TCD) It is a nondestructive universal detector. That has a moderate sensitivity. It is less satisfactory with carrier gas whose conductivities closely resemble those of most sample components. It measure change in thermal conductivity. b) Flame Ionization Detector (FID) It is destructive one. Combustible organic compound is needed for this type of detector. It is very sensitive and have wide dynamic range. The signal is depend on number of carbon atom in the organic analyte. It is mass sensitive instead of concentration sensitive. It is not sensitive to noncombustible analyte such as water and carbon dioxide gas.

c) Electron Capture Detector (ECD) It is non-destructive detector. It measures the changes in electron flown from source as eluting compound is capturing the electron. In the absence of organic species, the result is a constant standing of current. In the presence of organic molecules containing electronegative functional groups that tend to capture electrons, results in the current decreases markedly. 6. Signal processor and read out The data from the experiment will be displayed.It displays the peaks of all the substances in the sample. This is called the chromatogram. Software can perform all the calculations.

Application of Gas- Liquid Chromatography

GC-MS ANALYSIS OF ETHANOL EXTRACT FROM THE AERIAL PARTS OF FAGONIA LONGISPINA (FAMILY ZYGOPHYLLACEAE)

Apparatus and material: GC Clarus 500 Perkin Elmer equipment, Aerial parts of Fagonia longispina, 60% EtOH,
distilled water, hexane, ethyl ether, chloroform. (100% Dimethylpolysiloxane). 100%

Dimethylpolysiloxane

Method: Preparation of the extracts

1. 100 g dried powder of the aerial parts of Fagonia lonjispina extracted by using 60% EtOH. 2. The extraction is concentrated. Then diluted with water and partitioned with hexane, ethyle ether and Chloroform. 3. The residue of the hexane extract which is 2 gram, of the ethyl ether extract with 1.4 gram and of the 1.6 gram CHCl3 extract were performed using a gas chromatograph-mass spectrograph (GC-MS).
Gas Chromatography Mass Spectrometry Analysis

1. Analysis performed by using GC Clarus 500 Perkin Elmer equipment. 2. Compounds were separated on Elite-1 capillary column (100% Dimethylpolysiloxane).

3. Oven temperature was programmed at 100C for 1 min, then increased to 220C at the rate of 10C/min, then increased up to 260C at the rate of 5C/min which is for 9 min. 4. Ionization of the sample components was performed in the EI mode (70 eV). 5. The carrier gas was helium (1ml/min) and the sample injected was 2 l. 6. The detector was Mass detector turbo mass gold-Perkin Elmer. 7. The total running time for GC was 28 min and software used was Turbo mass 5.2. The individual constituents were identified by comparing their mass spectra with the spectra of known compounds stored in the spectral database,NBS,WILEYand NIST attached to the GC-MS instrument and reported.

Expected Results:

Figure 1: GC-MS Chromatogram of the hexane extract of the aerial parts of Fagonia lonjispina

Figure 2 GC-MS Chromatogram of the ethyl ether extract of the aerial parts of Fagonia lonjispina

Figure 3 GC-MS Chromatogram of the Chloroform extract of the aerial parts of Fagonia lonjispina

Conclusion: The analysis of the phytochemicals in the hexane, ethyl ether and chloroform extracts of Fagonia lonjispina aerial parts revealed 13 compounds namely that major players in antioxidant responses evoked by the plant.
List of compound present.

Ethyl Palmitate (26.71%), 9,12Octadecadienoic acid, ethyl ester( 16.03 %), 9.12.15 Octadecatrienoic acid,ethyl ester(z,z,z) (57.25%) , Phenol2,6-bis (1,1-dimethylethyl)4-methyl- ( 27.21%) , N-hexadecanoic acid( 12.24%), Tridecanoicacid ( 9.25%), 9,12-Octadecadienoicacid(z,z) ,methylester ( 8.16%), 11,14,17 Elcosatrienoicacid,methyl ester ( 34.69%) , Decanoic acid ( 12.24) , 9-Elcosene,( E ) ( 15.62%) , Cyclotetracosane ( 03.75%), 1-Heptadecene ( 23.12%), 1-Nonadecene ( 06.25%) .

UNIVERSITI TEKNOLOGI MARA SABAH FACULTY OF APPLIED SCIENCE

DIPLOMA IN SCIENCE
CHM260 CHAPTER 5 GAS-LIQUID CHROMATOGRAPHY
TITLE : GAS CHROMATOGRAPHY, COMPONENTS AND APPLICATION NAME : Ahmad Ismi Zulfadli Bin Ishak Matrix ID : 2010852022 GROUP : AS1206A1 DATE : 11th March 2013
LECTURER : MISS NOOR EZAWANIEE HJ MOULTON