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Agmatine determination in plant material: Aspects of the benzoylation and high performance liquid chromatographic analysis
Author(s) -
Wehr J. Bernhard
Publication year - 1995
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/pca.2800060504
Subject(s) - chemistry , agmatine , chromatography , reagent , derivative (finance) , degradation (telecommunications) , hydrolysis , yield (engineering) , organic chemistry , enzyme , putrescine , telecommunications , materials science , computer science , financial economics , economics , metallurgy
Various steps in the existing procedure (Slocum, R. D., Flores, H. E., Galston, A. W. and Weinstein, L. H., Plant Physiol. (1989) 89, 512) for the analysis of agmatine as its benzoyl derivative were found to impair the quantification of agmatine, leading to irreproducible results. Degradation of benzoylated agmatine is a major factor contributing to erratic results, and two mechanisms of degradation have been observed. Firstly, benzoylagmatine undergoes a temperature‐dependent hydrolysis in alkaline solution and this reaction has a half‐life of 168 min at 22°C. Therefore, the benzoylation reaction time and temperature have to be controlled in order to ensure reliable results; a maximum yield of benzoylagmatine may be obtained after incubation for 30 min. Secondly, benzoylagmatine can undergo rapid alkali‐catalysed methanolysis with a half‐life of 12 min at 22°C. Methanolysis produces a degradation product that was originally considered to be agmatine or benzoylurea, but is probably N ‐benzoylmethoxybenzamidomethylene imine. The formation of benzoylagmatine depends on the amount of benzoylchloride added since small volumes of the reagent do not exhaustively derivatize agmatine, hence leading to unreliable results. Mass spectrometry of benzoylagmatine showed that tribenzoylagmatine is the main derivative formed. The suggested modifications to the existing procedure enable the rapid and reliable determination of agmatine in various plant materials.