High-performance liquid chromatographic determination of polyamines in milk as their 9-fluorenylmethoxycarbonyl...

A high-performance liquid chromatographic method for the determination of polyamines in milk is milk is described. Polyamines were extracted in perchloric acid and derivatized with 9-fluorenylmethoxycarbonyl chloride (FMOC-Cl). The excess of reagent was reacted with aspartic acid before the analysis on a column-switching system. Linearity of derivatization was calculated for each amine and the coefficient of regression ranged from 0.994 to 0.999. Chromatographic separation of FMOC-polyamines was achieved with a gradient elution programme of water-acetonitrile. The correlation coefficients of the standard curves in the concentration range from 0.5 to 5 nmol ml-1 were higher than 0.991. The repeatability of the method, expressed as R.S.D. for each polyamines ranged from 3.0 to 8.6%. The percent mean recoveries at 1 nmol ml-1 spiking level were 49 +/- 3, 58 +/- 5, 61 +/- 5 and 48 +/- 4 for putrescine, cadaverine, spermidine and spermine, respectively. The limit of detection, calculated on the basis of three times signal-to-noise ratio, was 50 pmol ml-1 for each polyamine.