HPLC-ELSD测定草石蚕低聚糖含量的研究

Research on Determination of Oligosaccharide in Stachys sieboldii Miq.by HPLC-ELSD

建立了高效液相色谱法(HPLC-ELSD)测定草石蚕中低聚糖含量的方法。采用蒸发光检测器(ELSD),色谱柱为Waters XBridge Amide色谱柱(4.6 mm×250 mm,3.5μm),流动相为乙腈(A)-水(B),分别添加0.2%(V/V)三乙胺,梯度洗脱(A%为70%→45%→70%→70%,相应时间周期为0min→16min→16.01min→20min),流速1.0mL/min,柱温35℃。结果表明:蔗糖、棉籽糖、水苏糖在0.1~3.0g/L质量浓度范围内线性关系良好,相关系数不小于0.9986;相对标准偏差(RSD)为1.49%~1.67%(n=5);平均回收率分别为97.78%,97.99%,97.85%。干基草石蚕中蔗糖、棉籽糖、水苏糖含量分别为(6.38±0.10)%,(2.72±0.08)%,(43.38±0.25)%。

Establish a HPLC-ELSD method for the determination of oligosaccharide in Stachys sieboldii Miq.. The sample is separated on a Waters XBridge Amide column（4.6 mm×250 mm,3.5 μm）at 35℃ by ELSD and eluted with acetonitrile （A）-aqueous （B） solution added 0. 2% （V/V） triethylamine respectively, gradient elution（A% is 70%→45%→70%→70%, the corresponding time period is 0 min→16 min→16. 01 min→ 20 min）at a flow rate of 1.0 mL/min. The results show that there is a good linear range for sucrose,raffinose and stachyose in the quality concentration range of 0. 1-3. 0 g/L with a correlation coefficient no less than 0. 9986, the relative standard deviations are 1.49%-1.67% （n= 5）, and the average recoveries are 97.78%, 97.99% and 97.85% respectively. The sucrose, raffinose and stachyose in Stachys sieboldii Miq. are （6.38 ± 0. 10）%, （2.72±0.08）% and （43.38±0. 25）% respectively.