Improved high-performance liquid chromatographic method for the determination of polyamines as their benzoylated derivatives: application to P388 cancer cells.
A simple reversed-phase HPLC method was developed for the determination of eight polyamines or monoacetylpolyamines, as their benzoylated derivatives. Interfering products, inherent to the benzoyl chloride derivatization technique (benzoic acid, methyl benzoate and benzoic anhydride), were identified. A new derivatization procedure for their total elimination was developed without any loss of sensitivity and selectivity. Not only the HPLC method was validated, but also the choice of an internal standard was investigated. The results show that it is possible to use this HPLC assay to determine the polyamine content in P388 cancer cells. Furthermore, the method is now being used to evaluate the uptake of various polyamines by P388 cancer cells and by other cancer and parasitic cells.
Schenkel
E
E
Laboratoire de Toxicologie et de Chimie bioanalytique, Université Libre de Bruxelles, Belgium.
Berlaimont
V
V
Dubois
J
J
Helson-Cambier
M
M
Hanocq
M
M
D000818:Animals; D001565:Benzoates / Q000032:analysis* / Q000737:chemistry; D019817:Benzoic Acid; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007700:Kinetics; D007941:Leukemia P388; D011073:Polyamines / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results; D012996:Solutions; D013056:Spectrophotometry, Ultraviolet; D014407:Tumor Cells, Cultured
D001565:Benzoates; D011073:Polyamines; D012996:Solutions; D019817:Benzoic Acid