Re: AAA hydrolysis

tandersen@ccgateway.amc.edu
Thu, 21 Jan 99 14:35:27 EST

Cameron-

When running hydrolysis as you do, followed by Pico-Tag analysis, I
encountered the black tar associated with high carbohydrate
concentrations in the sample. I proceeded as normal with the
derivitization and chromatography and generally obtained excellent
results. The tar is quite insoluble and does not interfer too much
with analysis. I have less information about recovery of amino acids.
I suppose prudence would compel one to be sure the gaurd column is
functional, but the tar may not be a dead end.

Tom Andersen

______________________________ Reply Separator _________________________________
Subject: AAA hydrolysis
Author: cameron.olbert@arcb.us.astra.com at Internet-Mail
Date: 1/20/99 3:30 PM

Dear ABRF members,
We are currently trying to do AAA on a protein in a 10mM Tris 1%
n-Octyl-Glucopyranoside buffer and at the end of the hydrolysis step we are
seeing a black mass coating the bottom of the sample tubes. We are running
a vapor phase hydrolysis with 6N HCl 1% phenol, at 110C for 20 hours,
purging the reaction vial with nitrogen before hydrolysis. We believe the
black mass is caused by the nOG in the buffer (we've seen it in a buffer
blank). Any suggestions on how to avoid this hydrolysis problem are greatly
appreciated.

Thank you in advance
Cameron Olbert
Astra Research Center Boston
cameron.olbert@arcb.us.astra.com