Chromatography is one of several separation techniques that is defined as differential migration from a narrow initial zone. Electrophoresis is another member of this group. In this case, the driving force is the electric field, which gives different forces to the solutes with different ionic charges. The resistive force is the viscosity of the solvent not flowing.
The combination of these forces results in typical ionic mobility for each solute. Chromatography has many applications in the fields of biology and chemistry. This is widely used in biochemical research for the separation and identification of compounds in the sense of organic chemistry derived from biologics. In the petroleum industry, this technique is used to analyze complex hydrocarbon blends.
table of contents
- Benefits of Chromatography
- Purification from the results of chemical synthesis reactions
- Isolation of active compounds in natural ingredients
- Analysis of environmental waste
- Protein Separation
- Analysis and quality check in the food industry
- Application in the pharmaceutical industry
- Forensic Testing
- Drug testing
- Horse meat scandal (Horsemeat)
- Ebola immunization
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- Benefits of Chromatography
Chromatography is a laboratory technique for separating mixtures. Various mixed constituents move at different speeds, causing them to separate. This separation is based on the differential division between mobile and stationary phases. Subtle differences in the partition coefficients of compounds produce differential retention in the stationary phase and thus affect separation.
Chromatography can be preparative or analytic. The purpose of preparative chromatography is to separate the mixture components for later use, and thus is a form of purification.
Analytical chromatography is carried out normally with smaller amounts of material and is used to determine the presence or measure of the relative proportion of analytes in a mixture. The two are not mutually exclusive.
Chromatography was first used in Russia by Italian-born scientist Mikhail Tsvet in 1900. He continued to work with chromatography in the first decade of the 20th century, mainly for the separation of plant pigments such as chlorophyll, carotene and xanthophyll. A new type of chromatography developed during the 1930s and 1940s made this technique useful for many separation processes.
Chromatographic techniques developed substantially as the work of Archer John Porter Martin and Richard Laurence Millington Synge during the 1940s and 1950s, where they won the Nobel Prize in Chemistry in 1952.
They established the basic principles and techniques of partition chromatography, and their work led to the rapid development of several chromatographic methods: paper chromatography, gas chromatography, and what came to be known as high-performance liquid chromatography.
Since then, the technology has advanced rapidly. The researchers found that the main principles of Tsvet chromatography can be applied in many different ways, producing a variety of chromatographic varieties.
Progress continues to improve the technical performance of chromatography, allowing increasingly similar molecular separations. Chromatography has also been used as a method for testing the potential of cannabis.
Definition of Chromatography
Chromatography is a technique for separating components, or solutes, from a mixture based on the relative amounts of each solute distributed between the flow of moving fluid, called the mobile phase, and the adjacent stationary phase. The mobile phase can be liquid or gas, while the stationary phase is a solid or liquid substance.
The principle of chromatography is that chromatography exploits differences in the polarity of different molecules in a mixture. In this technique, the liquid acts as a mobile phase and sweeps away layers of particles called the stationary phase. A sample solution containing the mixture to be separated is put into a mobile phase which moves through a stationary phase.
The components in the mixture are separated based on their relative affinity for the two phases. Molecules that have greater affinity for stationary phases are slower than molecules that have lower affinity. The separated molecules are then compared with known and identified standards.
Chromatography has several advantages, including:
- Proper separation, analysis and purification is possible using chromatography.
- This requires a very low sample volume.
- It works on a variety of samples including medicine, food particles, plastics, pesticides, air and water samples, and tissue extracts.
- Mixture components separated by chromatography can be collected separately.
- Can be used to separate very complex mixes.
Definition of Chromatography According to Experts
The definition of chromatography according to experts, among others:
Chromatography can be defined as a physical mixture separation technique based on differences in the distribution of the components of the mixture between two phases, the stationary phase (solid or liquid) and the mobile phase (liquid or gas).
IUPAC (International Union of Pure and Applied Chemistry)
Chromatography can be defined as a method used to separate components in a sample, where the components are distributed between two phases, namely the stationary phase and the mobile phase. The stationary phase can be a solid or a liquid superimposed on a solid or gel.
Benefits of Chromatography
The following are a variety of benefits or the use of chromatography in general, including:
1. Purification from the results of chemical synthesis reactions
In organic chemistry we know the meaning of chemical synthesis reactions that allow the formation of products from certain reactants. In the synthesis reaction, it does not always produce a pure product or only one type of product, but also sometimes there are other products called by-products of the reaction product.
To separate the by-products from the main product, purification must be carried out. The easiest way is to use column chromatography. Column chromatography is based on differences in the polarity of the separated compounds so that the pure reaction compounds can be produced.
2. Isolation of active compounds in natural ingredients
We often use active compounds from certain natural ingredients because they have pharmacological activities such as antibacterial, antifungal, antioxidant, and others. To isolate these compounds purely from a natural material such as leaves, a separation method is needed, such as by using column chromatography.
With chromatography techniques, natural material samples will be extracted and then separated by elution. The result will be obtained the desired active compound and can be further characterized using more complex tools such as using FTIR or NMR.
3. Analysis of environmental waste
Chromatography plays an important role in many pharmaceutical industries and also in the chemical and food industries. Environmental testing laboratories generally want to identify small amounts of contaminants such as PCBs in waste oil, and pesticides.
The Environmental Protection Agency makes chromatographic methods to test drinking water and monitor air quality. The pharmaceutical industry uses this method to prepare highly purified ingredients in large quantities, and also to analyze purified compounds to track contaminants.
4. Protein Separation
Other applications of chromatography, especially HPLC ( High performance liquid chromatography ) are used to separate biochemical mixtures based on specific affinity between different cognitive components such as enzymes and substrates, antigens and antibodies, or receptors and ligands
In HPLC, the sample mixture is put into a mobile phase (often in the form of a solvent) and this is then pumped into a dense analytical column at high pressure for rapid separation of sample molecules. This is also called high pressure liquid chromatography.
5. Analysis and quality check in the food industry
Chromatography is used for analysis and quality checking in the food industry, with the separation and analysis of additives, preservatives, vitamins, and protein meanings . It can also be used to detect poisons and contaminants in food.
6. Application in the pharmaceutical industry
In chromatography it is used to purify materials and analyze chemical compounds to track contaminants, as well as to separate chiral compounds.
7. Forensic Testing
Gas chromatography is often used to investigate criminal cases. This can be a crime scene test (analysis of blood or cloth samples), verification of combustion (identifying the meaning of the chemicals responsible for fire to see if there is a foul play) or blood testing after death to determine levels of alcohol, drugs or toxic substances in the body .
8. Drug testing
Of course, not all blood samples are taken after death. The accuracy of chromatography can identify substances in the bloodstream making it valuable in testing for doping or drugs that improve performance in athletes as well.
Interestingly, in a story, doping tests also worked on horses, revealing how a new form of liquid hybrid chromatography combined with mass spectrometry can also be applied to horses.
9. Horse meat scandal (Horsemeat)
Speaking of horses, the 2013 scandal highlighted the fact that some meat traders were not fit to replace beef bread with horse meat. Because traditional methods for meat detection and analysis have proven to be inconclusive, chromatography is used to separate the two meats.
10. Ebola immunization
It turns out that chromatography is also important in saving millions of lives. The deadly Ebola virus, which has claimed more than 5,000 lives since it erupted late last year, has caused panic in the media and in the countries of Sierra Leone, Guinea and Liberia, where most of the virus is locked up.
When scientists try to fight this disease, chromatography has revealed itself to be very useful in determining which antibodies are more effective in neutralizing Ebola.
Although no drug has been conclusively validated, this technique plays an important role in the development of Zmapp’s experimental immunization and will continue to be used in ongoing research.