Dialysis overnight at 4 in PBS

what stationary phase did u use in the column chromatography?? did u try using different resins dedicated for the purpose of preliminary purification ..and then u can go for compound purification using C-18 column packing.

though it is slow but an effective column packing in purification,

I've used a conventional glass column with 60-120 mesh silica!

Dear Imran,

The best collumn will depend of the sample's characteristictic and the kind of impurities that you wanna to remove. Mayyou give some characteristic of the sample and impurities?

ya i am agreed with Rodrigo Salazar suggestion!!! its depands on characteristictic property.

60-120 mesh silica is not enough to get the level of purity you want.. U have to use the resins to get better result.. Choosing the resins will depend upon the type of compound you want to purify.

Can you specify the class of compounds?

Yes.. you r right Dr Ashrey. Resins are good option for peliminary purification of impurities viz. XAD-4, 8, 16 etc, HP-20 and so on....

Gudluck

:

You can also use preparative TLC or try with a Sephadex column, but the method depend on the type of compound you want to purify as Dr. Gupta said.

I don't recommend to use of preparative TLC method if your compound is UV-inactive,

1- First you should try COLUMN CHROMATOGRAPHY again on using different SOLVENT SYSTEM (Which solvent system you have used you didn't mentioned).

2- You should use SEPHADEX COLUMN (suggested by Gloria) or

3- go on HPLC using reverse-phase column (I recommend H80). Before using the HPLC you are to check the best solvent system on REVERSE PHASE TLC.

THIRD choice is much better if you have facility of HPLC.

you can also try with Sephadex column using Methanol:Water, 1:1

You can use Prep HPLC but that will surely be costly!!!! you can go for Column chromatography using resins (XAD-16) is good for polyphenols . or you can select a variety of resins as per your requirement.

You can also view this site:

http://chem.rochester.edu/~nvd/columntroubleshoot.html

Thanking all for your valuable suggestions and links.

Thanks Saikiran

Thanks Carlos

Crystallization or chromatography

Resins can be used to remove impurities.