植物组织中低聚糖乙酰化及毛细管气相色谱分析

Capillary Gas Chromatographic Analysis of Oligosaccharide Derivatives in Plant Tissue

建立一种用乙酰化衍生处理低聚糖并用毛细管气相色谱-FID进行分析的方法。以1-甲基咪唑为催化剂并以乙酸酐为乙酰化试剂,同时对植物样品中蔗糖、棉子糖和水苏糖等低聚糖乙酰化产物进行毛细管气相色谱分离和FID检测。确定了低聚糖乙酰化衍生物的毛细管气相色谱分析条件,并对低聚糖乙酰化反应条件及色谱分离条件进行了优化。结果表明,在80–1000ng·μL–1范围内线性关系良好,蔗糖、棉子糖和水苏糖的相关系数(R)分别为0.9952、0.9957和0.9877,并且精准度与回收率均较高。使用该方法对低聚糖进行乙酰化反应重现性好、所需样品材料及试剂量少且污染毒害小,能够得到理想的分离、检测和定量分析效果,适用于少量植物组织中低聚糖的定量分析。该方法在食品、医药检测和基础科学研究领域均具有广泛的适用性及参考价值。

We introduce the establishment and optimization of capillary gas chromatography-flame ionization detection（FID） for analysis of acetylated oligosaccharides.Chemical and instrumental analysis of oligosaccharides requires derivating them into their derivatives.We used 1-methylimidazole as a catalyst and acetic anhydride as a reagent for acetylation of oligosaccharides.The acetylation and separation of acetylated products of sucrose,raffinose and stachyose were optimized and effectively separated by gas chromatography on a DB-15 capillary column and detected by an FID detector.The method is accurate,precise and has high recovery rate.The recovery rates for sucrose,raffinose and stachyose were all above 90%.The standard curves of oligosaccharide acetylated derivatives showed a high linearity in the range of 80 to 1 000 ng·μL-1 concentrations.The linear correlation coefficients were 0.995 2 for sucrose,0.995 7 for raffinose and 0.987 7 for stachyose.The method is suitable for analysis of target compounds in the food sciences,medicine and biological sciences,with the advantages of high reproducibility,minor sampling and chemical consumption and environmental friendliness.