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

• Published in: Journal of chromatography. B, Biomedical sciences and applications Vol. 700; no. 1; pp. 23 - 30
• Main Authors: Dorhout, Bernard, Kingma, Anneke W., de Hoog, Elly, Muskiet, Frits A.J.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 24.10.1997; Elsevier Science
• Subjects: 1,19-Bis(ethylamino)-5,10,15-triazanonadecane; Acetylation; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Chromatography, Gas; Fundamental and applied biological sciences. Psychology; Leukemia L1210 - metabolism; N-Acetylpolyamines; N1, N11-Bis(ethyl)norspermine; Non peptidic neurotransmitters, polyamines; Other biological molecules; Polyamines; Polyamines - analysis; Polyamines - metabolism; Propylamines - analysis; Propylamines - metabolism; Putrescine; Sensitivity and Specificity; Spermidine; Spermine; Spermine - analogs & derivatives; Spermine - analysis; Spermine - metabolism; Tumor Cells, Cultured; 1,19-Bis(ethylamino)-5,10,15-triazanonadecane; Spermine; N-Acetylpolyamines; N 1, N 11-Bis(ethyl)norspermine; Spermidine; Putrescine; Polyamines; Capillary column; L1210 cell line; Gas chromatography; Polyamine; Simultaneous measurement; Quantitative analysis
• ISSN: 0378-4347; 1387-2273
• DOI: 10.1016/S0378-4347(97)00294-6

We describe a method for the profiling of polyamines, N-acetylated polyamines and the polyamine analogues N 1, N 11-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 internal 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 C.V.s) for analysis of 1–2×10 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.