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 [l5N4]-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 lUM. 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.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR000954-20
Application #
5221773
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
20
Fiscal Year
1996
Total Cost
Indirect Cost
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