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[Arabidopsis] benzoylation of polyamines

Brian Cady via arab-gen%40net.bio.net (by briancady101 from yahoo.com)
Sun Jun 9 05:12:23 EST 2013


Could this have to do with phthalates in many pipette tip plastics contaminating your samples?
I faintly remember seemingly similar issues arose in auxin work mentioned in a GC & LCMS workshop I went to.

Brian



Message: 1
Date: Thu, 06 Jun 2013 12:55:44 -0400
From: "James J. Campanella" <campanellj from mail.montclair.edu>
Subject: [Arabidopsis] benzoylation of polyamines
To: methods from magpie.bio.indiana.edu, plantbio from magpie.bio.indiana.edu,
    arab-gen from magpie.bio.indiana.edu
Message-ID: <d8e384f35718c796.51b086d0 from montclair.edu>
Content-Type: text/plain; charset=windows-1252


I don� know if anyone out there has performed this
protocol, but I am looking for advice. We are trying to determine the
concentration of 


polyamines in some plant tissue samples using a
benzoyl-labelling process and HPLC. Unfortunately, we have run into some
problems and 


not even gotten beyond the first step of labeling standards.




Analyzing the benzoylated standards on a C18 column using
an isocratic flow of 52% acetonitrile at 1 ml/min, we found no retention 


time
differences between labelled putrescine, spermidine and spermine. We found the
same result using a 50% methanol buffer system. 


It is unclear to us whether we are
having problems with the HPLC or whether it is an issue with the labelling
process.  





I was worried that we were not
getting any labelling at all so we ran a sample of the benzoyl chloride on the
HPLC just to see what its retention 


time may be�nd we found multiple large
peaks. It� entirely unclear to me why a reagent like benzoyl chloride should
have such multiple peaks. 


Sigma-Aldrich would not admit there was anything wrong with the product and has not yet answered my grad student's inquiries after a week's

time.




We would appreciate any help or advice that anyone can give.


Jim Campanella

--

----------

James J. Campanella, PhD
Associate Professor,
Department of Biology
Montclair State University
1 Normal Avenue
Montclair, NJ 07043

Alternate email address: jcamp from alumni.uchicago.edu

Ph: 973-655-4097
Fax: 973-655-7047


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