Simultaneous determination of polyamines, N-acetylated polyamines and the polyamine analogues BE-3-3-3 and BE-4-4-4-4 by capillary gas chromatography with nitrogen-phosphorus detection.
We describe a method for the profiling of polyamines, N-acetylated polyamines and the polyamine analogues N1,N11-bis(ethyl)norspermine (BE-3-3-3) and 1,19-bis(ethylamino)-5,10,15-triazanonadecane (BE-4-4-4-4) in L1210 murine leukaemia cells by capillary gas chromatography with nitrogen-phosphorus detection. The method makes use of four intemal standards. Prepurification comprises deproteinization, isolation with Sep-Pak silica at pH 9.0, conversion to heptafluorobutyryl derivatives and postderivatization organic fluid extraction. Within- and between-series precisions (given as CV.s) for analysis of 1-2x10(6) cells were: putrescine 5.5 and 29.4%; spermidine 1.6 and 7.1%; and spermine 3.2 and 7.6%, respectively. Recoveries relative to the respective internal standards, were in the 70.6-104.7% range. Accuracy and precision of measurements of BE-4-4-4-4 can probably be improved by the introduction of a separate pentamine internal standard. We conclude that the method can be used for studying the effect of BE-3-3-3 and BE-4-4-4-4, and possibly their metabolites, on polyamine homeostasis (biosynthesis, retroconversion, transport, terminal catabolism) and polyamine function.
Dorhout
B
B
Central Laboratory for Clinical Chemistry, University Hospital Groningen, Netherlands.
Kingma
A W
AW
de Hoog
E
E
Muskiet
F A
FA
D000107:Acetylation; D000818:Animals; D002849:Chromatography, Gas; D007939:Leukemia L1210 / Q000378:metabolism; D011073:Polyamines / Q000032:analysis* / Q000378:metabolism; D011437:Propylamines / Q000032:analysis* / Q000378:metabolism; D012680:Sensitivity and Specificity; D013096:Spermine / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D014407:Tumor Cells, Cultured
D011073:Polyamines; D011437:Propylamines; C059685:N(1),N(11)-diethylnorspermine; C082768:BE 4-4-4-4; D013096:Spermine