Amino Acid Purification
Amino Acid Purification and Column Chromatography: Why They Are a Perfect Match
Amino acids are essential compounds found in our body, and our body produces some of them, yet some we need to replenish from the food we consume. These important organic compounds are essentially the major building blocks of proteins in living organisms. They help produce new cells and repair tissue.
When it comes to consuming food products that are good in amino acid profile, you need to make sure that the quality of amino acids used or produced is top notch. You can implement numerous different column chromatography techniques for efficient amino acid purification.
To obtain high-quality purification of amino acids, you can use ion-exchange chromatography, or the size-exclusion chromatography technique. In these techniques, the amino acid mixture is loaded into a column filled with silica gel that is best suitable for the chromatography to take place. The silica acts as an adsorbent and helps separate and purify the compounds and amino acids from impurities and other particles as it pass through the column.
Though the fact is that all proteins are essentially made up of amino acids, they do have different structures and compositions, according to the type of amino acid profile they are made up of.
So for the amino acid purification process, the amino acids are first dissolved in a solvent and this solution/mixture is then applied at the top of the column. Now, as the solution flows down the column gradually, different amino acids are adsorbed by the silica gel at different levels depending on their affinity, leading to amino acid purification.
As the solvent passes through the column, it is collected in fractions, and each fraction will contain a different set of amino acids but similar properties. This will also be void of any impurities as they are adsorbed in the column. The fractions can be sent through the column multiple times to obtain purified compounds with the help of ion-exchange chromatography or size-exclusion chromatography.
You can monitor the elution process efficiently by using various detectors like UV-spectrophotometers or fluorescence detectors. This can be used to gauge the purity level of each fraction during the elution phase, and if need be you can repeat the column chromatography process multiple times till you achieve the desired amino acid purification levels.
Once the samples and fractions are eluted out of the column, you can implement various techniques such as high-performance liquid chromatography, or mass spectrometry to analyze the fractions, and identify the proteins/amino acids that have be eluted.
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