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Subject: RE: Prot seq & hplc
Date: Thu, 17 May 2001 17:58:37 -0700
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John -
It may not be the same problem , but I had a similar situation, although not
nearly as bad, on the HP 1100 on my HP sequencer. The lamp on the VWD
detector had a lot of hours logged; replacing the lamp took care of the
problem.
Scott Leigh, PhD
Senior Scientist
Protein Chemistry and Analytical Development
sleigh@fibrogen.com
direct (650) 866-7391
main (650) 866-7200
fax (650) 866-7255
FibroGen
225 Gateway Blvd.
South San Francisco, CA 94080
-----Original Message-----
From: John.Holt@aventis.com [mailto:John.Holt@aventis.com]
Sent: Thursday, May 17, 2001 9:21 AM
To: Recipients of ABRF List
Subject: Prot seq & hplc
The blank chromatogram on the 1090 LC of our HP G1000 Sequencer System has
a slope from 0 to 6mAU over the needed part of the separation. This slope
appears to have increased steadily over a year or so. As others have
remarked, the price of this is loss of sensitivity, which is now nearer
10pmole than the 1pmole that was previously possible. If this were simple
rp HPLC, I would match the absorbances of mobile phases A and B
experimentally by adding TFA to A (currently, B absorbs more than A).
Can anyone tell me (a) if I'm right in thinking that the problem is no more
than absorbance mismatch, (b) if there's an acceptable fix (doesn't require
extensive re-tuning of PTH separation) and (c), if there's a better (more
inert, or MS-friendly) chromophore to use than TFA to match solvents in rp
HPLC?
John Holt
Analytical Sciences
Aventis Pasteur
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