采用对氨基苯甲酸乙酯为衍生化试剂,建立柱前衍生化反相高效液相色谱法(rP-HPlC)测定2-去氧葡萄糖的含量。结果表明,最适的衍生化条件为反应温度85℃,反应时间1.5H,衍生化试剂(AbEE)和单糖(2-dg/glu)的摩尔比为25∶1;色谱条件为色谱柱HyPErSIl OdS2(250MMx4.6MM,5μM),流动相为0.02%TfA-乙腈(80∶20,V/V),流速1.0Ml/MIn,波长307nM。本方法在1~700Mg/l浓度范围内与峰面积呈良好的线性关系(r=0.9999);定量限(S/n=10)及检出限(S/n=3)分别为450和100μg/l;平均加标回收率为101.7%;rSd为0.7%;日内及日间精密度均小于0.7%。表明本方法灵敏度高,测定结果准确,重复性好,可用于2-去氧葡萄糖样品的准确定量分析。A method was developed for the determination of 2-deoxyglucose(2-DG)by reversed phase high performance liquid chromatography(RP-HPLC) after pre-column derivated with p-aminobenzoic ethyl ester(ABEE).The results showed that the optimum derivation conditions were as follows: reaction temperature was 85 ℃,reaction time was 1.5 h and molar ratio of derivative reagent(ABEE) to monosaccharide(2-DG/Glu) was 25∶1.The derivative was analyzed on a Hypersil ODS 2(250 mm×4.6 mm,5 μ) column with a mixture of acetonitrile to 0.02% trifluoroacetic acid(TFA)(2∶80,V/V) as the mobile phase,at the flow rate of 1.0 mL/min and the detection wavelength at 307 nm.This method has a good linearity at the concentration from 1 mg/L to 700 mg/L(r=0.9999).The limit of detection was 100 μg/L while the quantitative limit was 450 μg/L.The average recovery of 2-DG was 101.7% and the relative standard deviation(RSD) was 0.7%.The inter-day and intra-day precision were both less than 0.7%,This method is of high sensitivity,precision and repetition,indicating that it is suitable for precise quantitative determination of 2-DG.海洋公益性行业科研专项经费基金(No.201005020);福建省科技重大专项经费基金(No.2010NZ0001-2);厦门市海洋与渔业局科技计划项目经费基金(厦海渔合[2010]9号)资助项
