Simultaneous fluorometric determination of intracellular polyamines separated by reversed-phase high-performance liquid chromatography.
A reversed-phase HPLC technique in combination with fluorescent detection is described for simultaneous quantification of the precolumn Dansyl derivatives of intracellular amines. The derivatives were stable for at least one week, kept protected from the light at -20 degrees C. The detection limit was between 1 and 5 pmol for all tested polyamines. Serotonin coeluted with tryptamine. The method has a very good reproducibility for both, retention times and chromatographic peak areas. The average recovery of standard amine solutions added to cellular extracts was estimated to be higher than 90%. The described method enables a rapid, reliable and reproducible quantification of biogenic and related polyamines in biological fluids and tissues.
Matés
J M
JM
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Málaga, Spain.
Márquez
J
J
García-Caballero
M
M
Núñez de Castro
I
I
Sánchez-Jiménez
F
F
D000818:Animals; D015317:Biogenic Polyamines / Q000378:metabolism*; D002286:Carcinoma, Ehrlich Tumor / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003619:Dansyl Compounds / Q000378:metabolism; D006636:Histamine Release; D012701:Serotonin / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D014407:Tumor Cells, Cultured / Q000378:metabolism
D015317:Biogenic Polyamines; D003619:Dansyl Compounds; D012701:Serotonin