By Shin on Wednesday, June 4, 2003 - 12:45 am:
These day, I take experiments to purify hyaluronic acid from fermentated broth with ODS column.
I want to purify with no buffer because if I use any buffer I have to remove it after purfication even though some reviews use 0.01M TBAP(tetrabutylammonium phosphate)/acetonitrile 83/17 as mobile phase
Can anybody give any advise?
how can I eiliminate this buffer after purification if it is impossible to purify with no buffer?
By Tom M. on Wednesday, June 4, 2003 - 08:35 am:
Shin,
The TBAP is just an ion pairing agent used to increase the retention of hyaluronic acid. Do a literature search, there are many methods for hyaluronic acid that do not use an ion pairing agent.
Also, search the web for desalting columns. Good luck.
By Chris Pohl on Wednesday, June 4, 2003 - 11:35 am:
I agree with Tom that your first choice should be to use reversed phase with ion suppression. You should be able to get away with using a volatile acid such as formic acid to suppress ionization and maximize retention. Afterward you can strip off the formic acid in a rotary evaporator.
Another option would be to use tetrabutylammonium formate as your ion pair reagent. I don't think you can buy this reagent but you can easily make it from tetrabutylammonium hydroxide and formic acid. If you use this reagent and then pass the column effluent through a column packed with strong acid cation exchange resin in the hydrogen form, you should be able to remove the tetrabutylammonium ion and you could then remove the formic acid in a rotary evaporator. You might need to make sure you have sufficient solvent in the cleanup step in order to prevent absorption of your target in the cleanup resin.