Quantitation of the putative neurotransmitter agmatine as the hexafluoroacetylacetonate derivative by stable isotope dilution gas chromatography and negative-ion chemical ionization mass spectrometry

Douglas Stickle, Alan Bohrer, Richard Berger, Jeremiah Morrissey, Saulo Klahr, John Turk

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

A method is described for detection and quantitation of agmatine [4- (aminobutyl)guanidine] by gas chromatography/negative-ion chemical ionization/mass spectrometry after derivatization with hexafluoroacetylacetone. The lower limit of detection of the derivative was about 25 fmol on-column. For quantitative studies of agmatine content in biological samples, a procedure utilizing an internal standard ([15N4]agmatine prepared from [15N4]arginine) and an extraction step had a lower limit of detection of about 15 pmol for total sample content. Agmatine content was measured in rat tissue samples and normalized to protein content. Kidney and spleen samples exhibited the greatest content of agmatine per unit protein mass but agmatine was also detected in pancreatic islets and brain regions (cerebellum and cerebral cortex). On the basis of these measurements, it is estimated that the pancreatic islet intracellular agmatine concentration may exceed 1 μM. The sensitive and highly specific means of detection and quantitation provided by mass spectrometry may be useful in investigating the physiological role of agmatine in mammalian systems.

Original languageEnglish
Pages (from-to)129-136
Number of pages8
JournalAnalytical Biochemistry
Volume238
Issue number2
DOIs
StatePublished - Jul 1 1996

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