柱前衍生-反相高效液相色谱法测定2-去氧葡萄糖的含量

Detection of 2-Deoxyglucose by Pre-column Derivation Reversed Phase High Performance Liquid Chromatography

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(250mm×4.6mm,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.