利用高效液相色谱法测定转基因甘蔗中的低聚果糖  

Determination of Froctooligosaccharides in Transgenic Sugarcane with 1-SST using High Performance Liquid Chromatography(HPLC)

作　　者：武媛丽[1,2] 杨本鹏[2] 蔡文伟[2] 张树珍[2] 

机构地区：[1]海南大学农学院,海南儋州571737 [2]中国热带农业科学院热带生物技术研究所,海南海口571101

出　　处：《热带生物学报》2011年第2期101-106,共6页Journal of Tropical Biology

基　　金：中央级公益性科研院所基本科研业务费(ITBBZD0724)

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0.999 3以上,在转化1-SST的甘蔗中能够检测到蔗糖:蔗糖-1-果糖基转移酶的催化产物蔗果三糖(GF2),用此方法也可以较为准确地检测甘蔗茎杆节间其他糖类的组分及含量,可以用于甘蔗中糖类的定性和定量检测。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.

关 键 词：高效液相色谱法 低聚果糖 果糖 葡萄糖 蔗糖 

分 类 号：Q534[生物学—生物化学]