Gas chromatography is one of the sophisticated models of chromatography. As the name indicates the system operates with the flow of gas in the procedure.
This type of chromatography was especially designed to evaluate volatile compounds like fatty acid compounds, essential oils (volatile oils) etc.
The system is cumbersome and also with delicate instrumentation. It is quite expensive in terms of instrumentation, its maintenance and even the operation costs are also very high. Further expert handling is recommended in its operation requires unlike other easy chromatography techniques.
The gas chromatography principle involves separation of components of the sample under test due to partition in between gaseous mobile phase and stationary liquid phase. The components partitioned into gas come out first while other come later.
Gas chromatography runs on the principle of partition chromatography for separation of components. In terms of stationary and mobile phases it is categorized under gas-liquid type of chromatography i.e stationary phase is a liquid layer supported over a stationary phase while the mobile phase is an inert and stable gas. Hence the perfect name as Gas Liquid chromatography (GLC).
How gas chromatography works:
The gas is set to flow at a constant rate from the cylinder on to the liquid layer impregnated on solid support in a column. The sample is injected into the injection point and is carried by the mobile gas into the column. Inside the column, the components get separated by differential partition in between the mobile phase gas and stationary phase liquid. The component that partitioned into gas comes out of the column first and is detected by detector. The one partitioned into liquid phase comes out later and is also detected. The recordings are displayed onto a computer software. From these peaks one can identify the components and also their concentration.
Gas chromatography method: Below is the video for the instrumentation and method simultaneously.
Gas chromatography instrumentation
The gas chromatography apparatus can be listed as
1. The mobile phase gas in a cylinder: The mobile phase is an inert gas or non reactive gases like nitrogen, helium & hydrogen will do. The gas is kept in a gas cylinder and its out flow is controlled by regulator. There is also a pressure gauze which indicates the speed of flow of gas into the column. Most commonly used gas is helium.
2. The injection system: This is present before column yet inside the thermal chamber to load sample under analysis into the system.
3. Column of gas chromatography . The gas chromatography column is a usually long (few meters like 3 to 6 mts) and coiled for accommodation into a small thermal chamber. The column is mostly made of steel or glass.
The gc columns are 3 types viz.
♠ Packed column. This is a column into which solid beads are packed. This column has advantages like efficient separation and precise readings.
♠ Tubular column. Here into a stainless steel hollow tube a thin layer of liquid is coated to act as a stationary phase. This column offers least resistance to flow of gas.
♠ Support coated tubular column. Here into stainless steel column a thin solid layer is coated on to which a thin layer of liquid stationary phase is present.
4. The Detector: is another vital component of the gas chromatography apparatus. GC detectors detect the isolated components and helps in identification and quantification of the sample. They are of 4 types of gc detectors like
♦ Thermal conductivity detector: Here there are two columns which have a conducting wire in between. The gas is allowed to pass through the two columns of detectors i.e to one the effluent from gas chromatography column and to other gas from the gas cylinder directly. Since the temperature of both gases is same, the thermal conduction is constant.
When the sample is injected into gas chromatography column. The effluent gas carries the sample components into the detector column. Since effluent gas is mixed with sample components there results in difference in thermal conductivity from prior one recording. This difference in conductivity is specific for the component analyses and is recorded for further comparison and identification of the components as well as their quantity.
♦ Flame ionization detectors: Here the sample components from effluent are ionized by subjecting to flame in a chamber. These ions raise upwards and are attracted towards anode or cathode based on charge on them. When they impinge on the electrodes, current is passed which is recorded. The strength and intensity of current depends on the sample and is specific.
♦ Argon ionization detector;These detectors are similar to flame ionization detectors with only difference that argon ion gas is used to ionize the sample molecules. The argon ion are obtained by reacting argon gas with radio active elements. Once argon ionizes they try to get back to stable state by either taking or giving electron from the sample components thus making sample molecules to ions for detection.
♦ Electron capture detector. etc.
5. The computer to record the analysed readings. This is connected with the detector and hence records the detector changes in reference to the flow of separated components from the exit of the column. The record is called gas chromatograph.
5. The thermal chamber to fix or maintain fixed temperature.
* Precolumn and post column treatment of sample (if necessary). This is done to modify the sample. The sample should be stable on heating and also be separated properly. For this precolumn derivatisation is used and for the sample be detected properly post -column derivitisation is done by somw chemical change preferably.
As further improvement in GC, the gas chromatography apparatus is fixed with Mass spectroscopy system (gc-ms) for better analysis of components in terms of their mass.