hi sir , pls see the attached file i think it is use full to you

If you know the structures of the compounds of your mixture, you may try to use suitable protecting groups that modify only certain compound(s) of your mixture and lead to more (un-)polar derivative(s) with a different Rf value and better separation via column chromatography and/or crystallisation. After purification you can remove the protecting group again in order to get the pure compound.

kuin,

you can send the extract to me , I will look over it get the result send it to your address

You can use preparative TLC to purify that particular spot with the particular system

Dear,

Your question is not specific so that the 

Without further details, I'm inclined to agree with the approach described by Thamere and Murad.  In addition, crystallization may be another option.

My advice is that, you should first start by splitting your plant extract using different solvent of different polarity and obtain fractions such as hexane, DCM, butanol and that of water (liquid-liquid separation method). After that, you can now run the column using a mixture of specific solvent such as hexane and DCM (play on their %) for the non-polar fraction. The mixture of hexane and chloroform for mid-polar fraction (DCM), and finally the mixture of chloroform and ethyl acetate for polar fraction (butanol). You should also check the ratio between your sample and silica when loading your column.

NB. This is possible if your extraction was made using a solvent that can extract both polar and non-polar compounds (methanol, ethanol).

Fisrtly you must find the good eluent which gives you separate spots, then try with preparative TLC.

Check your solvent selection for extraction. You could benefit from the use of mixtures of solvents and Not just one. Also check your solvent selection for TLC. Remember that solubility and clarity of separation depend on the affinity of different chemicals for different solvents .  

You can do TLC with different solvent system for first better resolution with good difference in Rf value and also confirm wheather UV active or not. Followed by go ahead for Preparative TLC with same solvent system that giving best resolution as well as distingushible Rf value. then start scratching your desired band definitely you will mark it under UV if UV sensitive. then you can collection your compound by redesolving the silica with your selected solvent system.Then go for Mass, HPLC and NMR.