Determination Of Polyamines In Seawater By High Performance Liquid Chromatographic

We developed a highly sensitive and accurate reversed-phase high-performance liquid chromatography (RP-HPLC) method to determine dissolved free polyamines (putrescine, spermidine and spermine) in seawater. This method is based on pre-column derivatization with dansyl chloride (DNS-Cl). The derivatives were separated on a ODS column (150mm×4.6mm i.d., 5μm particle size, Agilent). The gradient elution was performed with two mobile phases A (acetonitrile) and B (ammonium acetate) at a flow rate of 1.0mL/min. The column eluate was monitored by fluorescence detection (excitation, 340nm; emission, 515nm). The detection limits for putrescine, spermidine and spermine were 9.6×10-11, 2.8×10-10 and 1.0×10-10mol/L respectively. The reliability of the method was satisfactory in terms of linearity (R>0.99), precision (relative standard deviation below 2.5%) and recovery (from 84.9 to 110.9%). This analytical technique requires neither preliminary extraction, nor long multi-step procedures. This method can apply in the dissolved free polyamines (putrescine, spermidine and spermine) determination in seawater.Five micro-algae (Skeletonema costatum, Chaetoceros curvisetus, Chaetoceros debilis, Chaetoceros didymus and Phaeodactyum tricornutum) were cultured in the lab and four polyamine concentrations in cluture medium were measured by HPLC using DNS-Cl reagent. Free 2-phenylethylamine, putrescine and spermidine concentrations in seawater increased remarkably during the later phase of the five micro-algae.Dissolved free polyamines (2-phenylethylamine, putrescine, spermidine and spermine) of the East China Sea in May, 2009 were detected using HPLC method. The concentrations of dissolved free 2-phenylethylamine, putrescine, spermidine and spermine in surface seawater ranged from 4.25～276.64nmol/L, 0.12~112.67nmol/L, 1.15~19.50nmol/L, 2.82~10.70nmol/L respectively. 2-phenylethylamine and putrescine concentrations were much higher than spermine and spermidine. The concentrations of the four polyamines in the surface seawater of the section cb were higher than those in bottom seawater.We investigated the concentrations of polyamines in the surface, 10m depth and bottom seawater during diatom red-tide dispersional area in the East China Sea. The concentrations of dissolved free 2-phenylethylamine, putrescine, spermidine and spermine in seawater ranged from 27.54～75.96nmol/L, 14.50～59.56nmol/L, 0.11～0.73nmol/L, 3.30～6.71nmol/L respectively. Spermine and spermidine concentrations were much lower than 2-phenylethylamine and putrescine.