使用高效液相色谱法测定海水中3种游离态多胺(腐胺、亚精胺和精胺)。海水样品经HClO4溶液酸化,丹磺酰氯衍生化,ODS色谱柱(150mm×4.6mmi.d.,5μm)分离,以0.1mol/L乙酸铵和乙腈为流动相梯度淋洗,流速为1.0mL/min,荧光检测(激发波长:340nm;发射波长:515nm)。本实验中腐胺、亚精胺和精胺的检出限(S/N=3)分别为9.6×10-11,2.8×10-10和1.0×10-10mol/L。在1×10-9~1×10-7mol/L范围内,3种多胺的浓度和荧光信号值均呈良好的线性关系(R〉0.99)。海水样品添加浓度在5×10-9,1×10-8,2.5×10-8和5×10-8mol/L的回收率为84.9%~110.9%;衍生方法的相对标准偏差〈3.6%,本方法中多胺衍生物不需要用有机溶剂萃取,衍生后可直接进样分析,避免了萃取过程中多胺的损失,极大地提高了分析速度。本方法具有选择性高,样品用量少,灵敏度好等特点,适合海水中游离态腐胺、亚精胺和精胺的痕量分析。
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.