Isolation and Purification of Natural Compounds
Many different natural compounds that are usually derived from plants, microbes, and other sources, need to be isolated and purified from their mixtures before they are used in various industrial processes and applications. But sometimes, separating and purifying these important natural substances from complicated or heavy mixtures can be a challenging task. You can easily separate and purify various compounds from a mixture by implementing column chromatography techniques, as they are efficient, easy to perform, provide pure and high-quality separation results, and are the best way to separate or purify the compound while retaining their efficiency.
Advantages of Column Chromatography for Natural Product Isolation & Purification
There are many key advantages of using column chromatography for the isolation and purification of natural compounds as they provide efficient separation and purification of various different mixtures.
High Resolving Power: The column chromatography process can effectively separate many different complex mixtures of natural products based on their differences in polarity, size, and other properties. This allows for the efficient isolation of target compounds, even from crude plant or microbial extracts as each compound has a different polarity towards the material that is used in the process leading to purified compound separation.
Compatible With a Wide Range of Natural Products: You can use and implement the column chromatography technique to purify or isolate a wide range of different natural products and mixtures, including alkaloids, terpenoids, flavonoids, and many others. Here, the choice of the stationary and mobile phases that you will use can be optimized for the target compounds to get efficient separation.
Can Purify Both Solids and Oils: If you are looking to separate or purify different compounds, column chromatography can be used to purify both solid and liquid natural products, unlike techniques like recrystallization, which are mostly limited to only solid compounds.
Selective Separation of Compounds is Possible: Column chromatography allows for the selective separation of compounds based on their physical properties such as polarity, size, and charge, to be separated or purified efficiently from their mixtures. This selective separation properties is useful and important when dealing with complex mixtures found in natural products.
High Capacity Samples: It can handle or can be easily scaled to accommodate relatively large sample volumes, making it suitable for isolating bioactive compounds from raw materials or biological extracts when compared to the other techniques.
Cost-Effective: Adsorption column chromatography, especially if you are using silica gel as adsorbents, is a much more cost-effective and widely used option in the initial separation stage due to its simplicity of the process and affordability.
High Purification: The process can effectively remove many different impurities present in the mixture, allowing you to obtain highly purified natural products in their highly purified state.
Key Steps Involved in Isolating Natural Compounds Using Column Chromatography
Sample Preparation
The first important step in this process is the proper extraction and preparation of the natural product sample. This typically involves extraction of the target compounds from plant, microbial, or other natural sources using useful/appropriate solvents. Next, the simple extract goes through a process to increase its concentration and get rid of any unnecessary parts of the surrounding material. After that, the extract sticks to a solid material, such as silica gel, to be inserted into the column.
Column Chromatography
The prepared sample of the mixture is then loaded onto the chromatography column that is packed with a suitable stationary phase. Here, the selection of the stationary phase, such as silica gel, alumina, etc, is based on the polarity of the compounds to be separated and other properties of the target compounds. The column is evenly filled and balanced with the initial mobile phase. The sample extract is then gently placed into the column, and the column is rinsed with a precisely chosen mobile phase, typically beginning with a non-polar solvent and slowly becoming more polar.
Fraction Collection and Analysis
As the column is eluted, fractions are collected in test tubes. These fractions are then analyzed to identify the ones that actually contain the target natural compounds. You can easily monitor the fractions by implementing or using techniques like TLC, HPLC, GC, or other analytical methods that effectively allow for the combination of fractions containing the same compounds based on the analysis. It also may be required to further purify the mixed fractions, usually by going through the column chromatography process again, this time using a different stationary phase.
Identification of the Compounds
The final purified natural compounds that are obtained after the process are then subjected to various techniques for structural identification to determine the molecular weight and fragmentation pattern of the compounds.
Column chromatography is one of the important and advantageous techniques for the isolation and purification of natural compounds from complex plant, microbial, and other natural sources. The high resolving power, scalability, and compatibility with a wide range of natural product classes make it an ideal choice. We are the global manufacturers and suppliers of premium quality Silica Gel for use in the processes for the isolation and purification of natural compounds.
