利用高效液相色谱法测定转基因甘蔗中的低聚果糖

摘要

为了检测蔗糖-1-果糖基转移酶（1-SST）转化甘蔗的果聚糖种类和含量,本研究使用高效液相色谱（HPLC）建立了测定果糖、葡萄糖、蔗糖和3种低聚果糖（蔗果三糖、蔗果四糖和蔗果五糖）含量的两个体系。甘蔗糖类提取采用超声波辅助水浸提法,实验以乙腈和水配比（V乙腈∶V水=87∶13;V乙腈∶V水=75∶25）作为流动相,Hydersir NH2柱作为分析柱,流速1.0 mL.min-1,柱温30℃,检测器为示差折光检测器。结果表明,糖类质量浓度在0.625～50 g.L-1内呈现良好的线性关系,相关系数在0.9%In order to determine varieties and contents of froctooligosaccharides in transgenic sugarcane transformed with 1-SST.Two saccharides determination methods using HPLC were established,one for determining the fructose,glucose and sucrose,the other for determining three Froctooligosaccharides（GF2,GF3 and GF4）.Saccharides of sugarcane were extracted with water,using a mobile phase composed of acetonitrile and water（87/13 V/V;75/25 V/V） at a flow rate of 1.0 mL·min-1 for separating,Hydersir NH2 column（4.6 mm×250 mm,5 μm） temperature was 30 ℃,the differential refraction detector was used to detect samples.The results indicated that the linear ranges of the six saccharides were 0.625—50 mg·mL-1,with a correlation coefficient exceeding 0.999 3.kestose GF2 in sugarcane transformed with the 1-SST gene were detected.Our methods can be used for accurate qualitative and quantitative determination of fructose,glucose and sucrose in sugarcane.