Determination of erythrocytic polyamines by reversed-phase liquid chromatography

A simple and rapid semiautomated procedure for determining polyamines in erythrocytes by high-performance liquid chromatography is described. Putrescine, spermidine, and spermine are converted to fluorescent dansyl derivatives, extracted with cyclohexane, and separated in less than 10 min on a reversed-phase C18 ODS column, with an acetonitrile-water gradient as the mobile phase. The method showed a coefficient of variation of 2.73% for spermidine and 3.27% for spermine. The respective reference values, evaluated in 10 healthy patients, were 7.88 (SD 2.09) and 5.42 (SD 1.55) mumol/L of packed erythrocytes. Only negligible amounts of putrescine were found.