Determination of dansylated polyamines in red blood cells by liquid chromatography-tandem mass spectrometry.
The concentration of polyamines in red blood cells (RBCs) is considered to be an index of cell proliferation. This index has been demonstrated to be of clinical importance for the follow-up and treatment of some cancer patients. The concentration of polyamines in RBCs is usually determined by high-performance liquid chromatography (HPLC) with fluorescence detection. In the current work, we present a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of putrescine, spermidine, and spermine, the three major polyamines in RBCs. The polyamines were dansylated and analyzed by an LC gradient of 20-min duration on a C18 column on-line with a tandem mass spectrometer. An internal standard (1,8-diaminooctane) was used for quantification. This method exhibited excellent linearity for the three polyamines with regression coefficients higher than 0.99. The limits of detection for putrescine, spermidine, and spermine were 0.10, 0.75, and 0.50 pmol/ml, respectively. The intrarun precision values for putrescine, spermidine, and spermine all were better than 10%, and the interrun precision values were 13%, 9%, and 20%, respectively. The LC-MS/MS method is sufficiently simple and reliable enough to replace the currently used HPLC method with fluorescence detection in which putrescine is not always detectable.
Département de Biologie Intégrée, Centre Hospitalier Universitaire, 38043 Grenoble cedex, France. vducros@chu-grenoble.fr
Véronique
V
Ducros
Daniel
D
Ruffieux
Hélène
H
Belva-Besnet
Florence
F
de Fraipont
François
F
Berger
Alain
A
Favier
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003619:Dansyl Compounds / Q000737:chemistry*; D004912:Erythrocytes / Q000737:chemistry*; D006801:Humans; D011073:Polyamines / Q000032:analysis* / Q000737:chemistry; D011700:Putrescine / Q000032:analysis / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013095:Spermidine / Q000032:analysis / Q000737:chemistry; D013096:Spermine / Q000032:analysis / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*
D003619:Dansyl Compounds; D011073:Polyamines; D013096:Spermine; C029038:dansyl chloride; D013095:Spermidine; D011700:Putrescine