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Old 06-06-2013, 06:55 PM posted to bionet.plants
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Default benzoylation of polyamines


I donít 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 beóand we found multiple large
peaks. Itís 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:

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

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