研究高效液相色谱(HPLC)示差折光检测法测定甘蔗节间糖含量,建立一种简便、快速和准确可靠的,能同时测定果糖、葡萄糖、蔗糖含量的高效液相色谱法。HPLC方法采用Agilent Zorbaxcarbohydrate柱(4.6mm×250mm,5μm),流动相为乙腈-水(75:25,V/V),流速1.0mL/min,柱温30℃,检测器温度35℃。该法相对标准偏差(RSD)为0.82%~0.87%(n=5),加标回收率为97.84%~99.12%,糖类质量浓度在0.625~20mg/mL内呈现良好的线性关系,相关系数在0.9982(n=6)以上。采用本法测定两种甘蔗品种不同时期、不同节间糖含量,均获得满意结果。
Fructose,glucose and sucrose contents in sugarcane internodes were determined using high performance liquid chromatography with refractive index detector(RID).The chromatographic separation was achieved on an Agilent Zorbax carbohydrate column(4.6 mm×250 mm,5μm),which was held at a constant temperature of 30℃,using a mobile phase composed of acetonitrile and water(75:25,V/V)at a flow rate of 1.0 mL/min.RID temperature was set as 35℃.Average spike recoveries for 5 replicate determinations of three saccharides were found in a range of 97.84%to 99.12%,with a relative standard deviation(RSD)between 0.82%and 0.87%.The developed calibration curves displayed good linearity over a concentration range of 0.625 to 20 mg/mL,with a correlation coefficient exceeding 0.9982(n=6).This method has been applied to determine two different varieties sugarcane internodes with satisfying results.