Dear Abdulelah Ahmed Thabet

You can start with a liquid-liquid fractionation using less polar to more polar solvents (hexane-chloroform-ethyl acetate-butanol and the remaining in water)

1- re-suspend your extract in water

2-wash your suspension with hexane and separate in a funnel; and repeat until the hexane added remain clear before passing to chloroform... and so on

and then check each fraction for its polyphenol content, they should be present either in ethyl acetate and/or butanol fractions and in lesser amount  in the water fraction.

For column fractionation, you should standardize your solvent system yourself using analytic TLC. For this purpose you should choose a system of 2 or 3 miscible solvents (1 less polar and other (s) more polar) and check their ability to separate your compounds before going for the column.

Regards!

NOBOSSE

Dear Researcher,

Completely agree with Nobosse, While analyzing polyphenols you need some  methods.  I am attaching some useful documents here of  my respected colleges. Good luck

I am also agree with  Nobosse and Ajay . The main step after extraction is to isolate the fractions in pure form. This can be done by Column chromatography and TLC. The purity of compounds can be checked by HPLC. Good Luck!

I  think all are on the same wavelength ,   get the aqueous extract  , perform the TLC test on a  square type  ( 4 X 4 inches size ) to check  first on a one dimensional  side , the behaviour of the  extract contents , the the same plate can be eluted  on the second dimension . this will show us the no /  amount  of the impurities being  eluted in the first dimension.  The solvent system we just have to find out through the trial basis &  use that which  gives maximum no. of  compounds   on the TLC plate.  The Column chromatography  solvent system has to be   fairly stronger due to thickness of the   silica or cellulose column bed . Collect the column fractions &  do their  UV   , try to separate them as per their behaviour. Separate the one U R interested most , evaporate them for further study & c=hemical  characterixzations .

USE TLC to find out best solvent system for separation of your compounds

I fully agree with Nobosse.

I agree with the answers of previous researchers

you will find attached an article published recently explains the method of extraction of phenolic fractions by liquid-liquid extracion.

Dear all thanks so much for your response.

dEAR Abdulelah Ahmed Thabet

FIRST DESIGN BETTER TLC SOLVENT SYSTEM FOR NETTER SEPARATION THEN DO COLUMN CHROMATOGRAPHY IN BEST SOLVENT FOR BETTER SEPARATION.

Thanks.