Re: Peptide Content
Ioannis Papayannopoulos (iap@iname.com)
Wed, 22 Jul 1998 16:37:51 -0400
Perhaps I am missing something but why not use RP-HPLC with UV detection at
214 nm. The rule of thumb I remember from my graduate school days for
angiotensin I (decapeptide) was that 1 nmol of the peptide injected and
eluted off the column at 1 mL/min gave a response of about 0.1 AU at 214
nm. With state of the art HPLC instruments and detectors (microbore
columns, low flow rates, low volum flow cells with long light path lengths,
etc.) one should be able to quantitate in the 10-50 range, I should think.
HPLC obviously lends itself to this as it is easy to generate the standard
curves needed for quantitation, with autosamplers and the software which
most HPLC instrument vendors provide.
Ioannis Papayannopoulos
CytoMed, Inc.
Cambridge, MA
At 12:12 PM 7/22/98 -0600, Henry Wolfe wrote:
>I am looking for a sensitive, linear, quantitative metod for determining
>peptide content. The restrictions I am under are the following: I have a
>12-residue peptide with one primary amine, three disulfides and no
>aromatic amino acids. Also, I do not own an amino acid analyzer or
>protein sequencer, but have good UV, fluorescence, CE and HPLC
>equipment. Any suggestions? References to journal articles would be an
>added bonus.
>
>Thanks,
>
>Hank Wolfe
>henry.wolfe@nycomed-Amersham.com
>
>