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[Arabidopsis] benzoylation of polyamines

James J. Campanella via arab-gen%40net.bio.net (by campanellj from mail.montclair.edu)
Thu Jun 6 11:55:44 EST 2013


 =

I don=92t know if anyone out there has performed this
protocol=2C but I am looking for advice=2E We are trying to determine th=
e
concentration of =



polyamines in some plant tissue samples using a
benzoyl-labelling process and HPLC=2E Unfortunately=2C we have run into =
some
problems and =



not even gotten beyond the first step of labeling standards=2E



 =

Analyzing the benzoylated standards on a C18 column using
an isocratic flow of 52=25 acetonitrile at 1 ml/min=2C we found no reten=
tion =



time
differences between labelled putrescine=2C spermidine and spermine=2E We=
 found the
same result using a 50=25 methanol buffer system=2E =



It is unclear to us whether we are
having problems with the HPLC or whether it is an issue with the labelli=
ng
process=2E  =





 =

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=97and we found multiple large
peaks=2E It=92s entirely unclear to me why a reagent like benzoyl chlori=
de should
have such multiple peaks=2E =



Sigma-Aldrich would not admit there was anything wrong with the product =
and has not yet answered my grad student=27s inquiries after a week=27s

time=2E
 =




We would appreciate any help or advice that anyone can give=2E
 =


Jim Campanella

--

----------

James J=2E Campanella=2C PhD
Associate Professor=2C
Department of Biology
Montclair State University
1 Normal Avenue
Montclair=2C NJ 07043

Alternate email address=3A jcamp=40alumni=2Euchicago=2Eedu

Ph=3A 973-655-4097
Fax=3A 973-655-7047


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