Determination of Dissolved Free Putrescine,Spermidine and Spermine in Seawater by High Performance Liquid Chromatography

A highly sensitive and accurate reversed phase high performance liquid chromatographic (RP-HPLC) method was developed 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 (150 mm×4.6 mm i.d.,5 m particle size,Agilent) .The gradient elution was performed with two mobile phases A (acetonitrile) and B (ammonium acetate) at a flow rate of 1.0 mL/min.The column eluate was monitored by fluorescence detection (λex=340 nm; λem=515 nm) .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 (R0.99) , precision(relative standard deviation below 3.6% ) and recovery (from 84.9% to 110.9% ) .This analytical technique requires neither preliminary extraction,nor long multi-step procedures.This method can be applied to the determination of dissolved free polyamines (putrescine,spermidine and spermine) in seawater.