Guanidine HCl compatability of C18

[Amar]

Helo every body,
Is C18 RP-HPLC column compatible with Gunidine chloride? My peptide is insoluble without Gunidine. If not, which column can be used with denaturant?
Thanks
Amar

[MartinG]

I have been able to inject a protein in 6M GnHCl on a R1 POROS reversed phase column, which has a selectivity similar to a C4 column. The retention time was the same than when the same protein was injected without GnHCl. So it worth trying.

Good luck

Martin

[HW Mueller]

If one does a lit search on protein RPHPLC one sees a lot of applications with all kinds of columns using guanidine HCl. Having used only urea I would still venture a guess that the only cause for worry is an acidic pH, with all that Cl- around. On the latter: Check into all those chains about Cl-, HCl, and stainless steel.

[Amar]

Helo HW,
do you mean you have used urea on C18 RPHPLC columns? is it okay to use? coz I use either GuCl or urea.
Thanks for previous reply.
Amar

[Mark Tracy]

Urea won't hurt your column at all. Just be sure that the sample does not precipitate when it mixes with the mobile phase.

[Amar]

Hi Mark,
Thanks. That information was of great help. How about DTT? is it going to damage C18 column?
Amar

[Mark Tracy]

DTT is also harmless, so is iodoacetamide (at least harmless to your C18 column).

[Amar]

Hi mark, I really appreciate your answers. I will post if I have any question in future.
Amar

[HW Mueller]

Yes I did use C-18 columns with Urea, I didn´t mention it specifically as there are also sundry examples in the literature, and also because I can´t see any possibility for reaction. Again, the Cl- of guanidine should be watched if you use low pH, because of the stainless steel.