Cameron-
I too experience the black tar in certain hydrolyzed samples. Like Tom, I have
not found it to interfere with derivatization or chromotography. However, a
quick, low speed centrifuge will drop the tar from solution and away from your
injector needle, or allow you to transfer the sample away should you so desire.
Ryan Coleman.
______________________________ Reply Separator _________________________________
Subject: Re: AAA hydrolysis
Author: tandersen@ccgateway.amc.edu at smtpgate-zgi
Date: 01/21/1999 2:35 PM
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
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