HPLC-ELSD法检测葛根酶解物中纤维低聚糖的含量

摘要

建立高效液相色谱-蒸发光检测器(High Performance Liquid Chromatography-Evaporative Light Scattering Detection)测定葛根酶解制备物中葡萄糖、纤维二糖和纤维三糖的方法.HPLC条件为:采用Asahipak NH2P-50-4E(4.6 mm×250 mm,5μm)色谱柱,柱温35℃、流动相:乙腈:水=70:30(体积比)、流速0.8 mL/min;ELSD参数为:漂移管温度70℃,N2流速2.0 L/min.结果表明,葡萄糖、纤维二糖和纤维三糖的浓度与其峰面积在0.07 mg/mL~0.7 mg/mL内有良好的线性关系(R2>0.99),精密度RSD 0.99%~3.1%,平均回收率98.17%~103.42%.%A high performance liquid chromatography (HPLC) with evaporative light scattering detection (ELSD) method was developed for the dermination of 3 saccharides such as glucose,cellobiose and cellotriose in the enzymatic hydrolysate of Radix Puerariae. A Asahipak NH2P-50-4E(4.6 mm×250 mm,5μm)column was used and the HPLC contidions were as follows:colunmn temperature 35℃, mobile phase:acetonitrile:water=70:30, flow rate 0.8 mL/min . And also the ELSD conditions were:detector drift tube temparature 70℃, nitro-gen flow rate 2.0 L/min. The results showed that there were good linear correlations between the concentration and the peak area of the 3 kinds of saccharides with the detection limitation from 0.07 mg/mL to 0.7 mg/mL(R2>0.99).Precision RSDs of standard samples for the determination of the sugars were 0.99%-3.1%and average recoveries for the sugars in 5 real samples were 98.17%-103.42%.