I have already collected the total alkaloidal extract from crude by acid base extraction. How can I isolate all the alkaloids present in total alkaloidal extract?
You should select chromatographic material (aluminium oxide neutral, acid, etc, Sephadex) and technique (Chromatotron, column) according to the type of alkaloids. Literatures about your plant material will help you.
Using neutral alumina is a good chromatographic method, also you can use Sephadex columns.
You should select chromatographic material (aluminium oxide neutral, acid, etc, Sephadex) and technique (Chromatotron, column) according to the type of alkaloids. Literatures about your plant material will help you.
Column is the only solution. The more advanced technique you use, the better the result will be. Conventional column will also do but the suggestions in previous posts are far advanced and more productive too.
If you have a good TLC profile for the alkaloidal mixture, then you can go for prep-TLC and separate the individual band from the mixture and then try to get the crystals of the single compound.
Column purification remains the choice. For the fact that all the members of the mixture are alkaloids any other method chosen will be the same in terms of reactivity for them. However, their relative mobilities would autommatically differ due to variations in molar masses in a seperating column. It is advised that a basic stationary phase be used as adsorbent because an acidic type will lead to a chemical reaction between the labile phase (alkaloids) and the adsorbent. The net effect of this ugly development is tailing and non-seperation of the substances.
For separation of pure individual alkaloids, the crude alkaloidal extract need to be chromatographed through Column Chromatography using appropriate adsorbent silica gel, alumina, Sephadex etc.) and careful solvent elution followed by regular monitoring of eluents by TLC. Pooling together TLC homogenous eluents and crystallization will furnish pure alkaloids. For more precise separation of co-eluted compounds, HPLC would be the correct option.
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