I am starting research work on bee pollen, I want to extract the biochemical constituents of a pollen. I need to know the overall methodology used on this type of technique?
Methods quantification of biochemical constituents depends on type of constituent you want to determine.Most common method for extraction of biochemical constituents is methanolic extraction and can be determined by methods specific for the constituent.It is not clear which constituent you want to quantify.
I want a protein extraction method of pollen? because they present the exine and I need a method to destroy it before extracting proteins?
Why you want to remove exine?As it is very difficult to remove it due to small size of pollens.Pollen including exine can be used for protein extraction.
But with the exine can not easily extract the protein from the cell. have you an assisted, how to extract the proteins prior to the determination?
Ihitassen,
for better protein extraction from bee-pollen you can first extract lipid fraction by hexane or heptane (in sonic bath for 15 min). In that way you will desintegrate exine (at least partially if not totaly). After that you can extract other fraction together with 50% ethanol. After that extract polyphenols with ethyl-acetate from ethanolic solution. Then with buthanol you can extract protein fraction. At the end, in aquatic fraction sugars will remain.
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