Hi all friends
i am asking for a procedure for liquid- liquid fractionation for a polar(methanol extract) thermolabile plant extract , i want to fractionate the extract to 5-6 fractions to ease the isolation process
Im working separating enzymes using a PEG 8000/phosphate system. What do you want to know?
I am also doing a liquid liquid fraction....if u can tell me u are interested in separation of which type of compound i can help u out.....
hummm understand... well, the system consists in two miscible liquids into a solution that turns non miscible in certain conditions like concentration, pH, temperature. When it happens is generated a chemical gradient over the biphase system. This chemical gradient drives the necessary energy to make the separation of your interest substance.
The basics evolve the independent preparation of two solutions that are putted together to a certain condition be reached. Some people use two polymeric solutions, another ones uses a polymer and a salt solution. There another possible combinations. Once the volume of these solutions are calculated you put them together and complete the final volume and incubate your new solution in a certain temperature.
Summarily is this... But is necessary a standardization of your chemical system before using it as strategy to separate your flavonoids as well.
depends on the type of system that you want to use .Do you want to evaluate what combination is better to perform your purification procedure testing different kinds of extraction combined to fractionation? If positive, you might decide what kind of system that you want to use.
Im using a kind of system that uses PEG 8000 and phosphate buffer solutions. In my case, for proteins per example, i built the system and put on it around to 2mg (i dont remember the exact value here now) of my interest protein dissolved in water. Depending on your chosen system, these extracts with organic solvents that your sample is dissolved can disturb the phase formation of your fractionation system. Generally concentrated samples were preferred to my (enzymes) case. Well, dont worry.
Decide firslty checking the literature the best fractionation system reported to flavonoids dissolved in these solvent (or something similar) to starting using it. This is what I will do.
hey if u r interested in a alkaloid extraction with difeerent solvents system .... then u can first mix the methanol extract with water and petroleum ether in equal amount n fractionate it... the ether layer will contain waxes, terpeoinds, the aqueous phase will contain amino acids, alkloids, sugar molecule. then acidify with .2 N HCl then extract with chloroform the aqueous phase is basified with ammonia till ph 11 then again extract with chloroform the chloroform phase will contain alkaloids. for more information u can refer Harbone - phytochemical extraction
Lincoln Scott could you please send me the paper ref. where this method has been used.
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