Has anyone tried using a nitrogen specific HPLC detector for peptide
quantitation?
Antek (http://www.antekhou.com/product/chrom/HPLC.htm) makes one, for
example.
I don't have any experience with this type of detector but was wondering if
anyone
else on the list has tried quantitation of peptides with this approach?
I've run across this problem of performing absolute quantitation of
peptides and proteins
before and have thought of this approach. It would be nice to know if this
works.
David Bunk
Research Chemist
Analytical Chemistry Division
National Institute of Standards and Technology
At 06:05 PM 5/30/00 +0200, mraida@gmx.de wrote:
>Dear Alessandro,
>after long years working with peptides, I am sorry to say that amino acid
>analysis is the only reliable method, in my eyes. The drawbacks beside the
>work are either amino acids which are destroyed or amino acids which are
>generally to high, but in case of a known sequence - this should be with a
>pure
>synthetic peptide - you can look for the most stable amino acids. There is
>still a small error but less than any other method. I used 160 ƒC vapour
>phase hydrolysation and OPA/FmoC precolumn derivatisation on a HP Aminoquant
>with RP separation and either UV or fluoresecence detection, standard
>protocolls. In comparison to other methods (Ninhydrin etc.) the results
>were within
>small deviations, even if the whole procedure had been carried out in a
>different lab on the same peptide.
>Yours
>Manfred Raida
>APB Proteomics
>
>--
>Dr. Manfred Raida
>European Proteomics Lab
>Amersham Pharmacia Biotech
>Munzingerstrasse 9
>D-79111 Freiburg
>Germany
>Email: Manfred.Raida@eu.apbiotech.com
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