Determination of rat hepatic polyamines by electron-capture gas chromatography.
BACKGROUND
An efficient and reproducible electron-capture gas chromatographic protocol that allows the simultaneous detection and quantification of the polyamines putrescine (1,4-diaminobutane), cadaverine (1,5-diaminopentane), and spermidine (N-[3-aminopropyl]-1,4-diaminobutane) was developed.


METHODS
Hepatic tissue from male Sprague-Dawley rats was used for analysis. The polyamines and the internal standard (sertraline) were extracted and derivatized with pentafluorobenzoyl chloride (PFBC) under basic aqueous conditions prior to analysis on a gas chromatograph equipped with a capillary column (narrow-bore fused silica column; 25 mm x 0.32 mm) and an electron-capture detector.


RESULTS
PFBC reacts with the amine functions of the polyamines examined here to produce PFB derivatives with high sensitivity on electron-capture detection. The method permitted the quantitative analyses of all three amines in rat hepatic tissue; the concentration of putrescine, but not spermidine, was increased significantly following a 14-day administration of the diamine oxidase (DAO) inhibitor aminoguanidine. Cadaverine was also present at increased concentrations in hepatic homogenates from aminoguanidine-treated rats.


CONCLUSIONS
Extractive derivatization with PFBC followed by gas chromatographic analysis using electron-capture detection results in a rapid and reproducible assay that permits the simultaneous detection and quantification of putrescine, cadaverine, and spermidine in biological tissue.
Clements
Robert L H
RL
Department of Psychiatry, University of Alberta, Edmonton, Alberta, Canada.
Holt
Andrew
A
Gordon
Emily S
ES
Todd
Kathryn G
KG
Baker
Glen B
GB
D000818:Animals; D001565:Benzoates; D002103:Cadaverine / Q000032:analysis*; D002849:Chromatography, Gas / Q000379:methods*; D006151:Guanosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D008099:Liver / Q000737:chemistry*; D008297:Male; D011700:Putrescine / Q000032:analysis*; D051381:Rats; D020280:Sertraline; D013095:Spermidine / Q000032:analysis*; D005464:Fluorobenzenes; D005466:Fluorocarbons
D001565:Benzoates; D002103:Cadaverine; D006151:Guanosine; D011700:Putrescine; D020280:Sertraline; D013095:Spermidine; C402368:N(7)-aminoguanosine; C014737:pentafluorobenzoyl chloride; D005464:Fluorobenzenes; D005466:Fluorocarbons