HPLC-ELSD法测定甘蔗叶片蔗糖含量

Determination of Sucrose Content in Sugarcane Leaves by High Performance Liquid Chromatography with Evaporative Light Scattering Detector(HPLC-ELSD)

利用水提法提取甘蔗叶片中的糖类,采用Xbridge NH2色谱柱,流动相为乙腈∶水（70∶30,V/V）加0.1%NH4OH,流速1.0 mL/min,柱温25.0℃,建立了一种高效、快速的测定甘蔗叶片中蔗糖含量的高效液相色谱——蒸发光散射检测法（High Performance Liquid Chromatography with Evaporative Light Scattering Detector,HPLC-ELSD）。在该方法中,ELSD检测器漂移管温度为85.0℃,氮气流速为2.0 L/min,增益为2。在以上条件下,蔗糖标准品能与糖类标准品中的其它糖类明显分开,甘蔗叶片中蔗糖也能与其它糖类明显分开,分离效果均良好。蔗糖浓度为0.2~0.9 mg/mL时,呈良好的线性关系（R2=0.994 4）。本方法精密度达到2.0%,表明仪器的精密度良好;蔗糖回收率为99.7%,说明本方法的回收率良好;通过间隔一定时间多次试验,测定蔗糖在提取液中的稳定性,结果表明多次试验误差为0.9%,表明此样品抽提法获得的样品较为稳定。

A highly efficient and rapid method to determine the sucrose content in sugarcane leaves was established. The sample was extracted by water, separated by the XBridge-NH2 chromatographic column, with acetonitrile：water（70：30）supplemented with 0.1% NH4OH as the mobile phase. The flow rate was 1.0 mL/min, and the column temperature was 25.0℃, while the ELSD drift tube temperature was set to 85.0 ℃, and the nitrogen flow velocity was 2.0 L/min. Under the above condition, sucrose, glucose and fructose in both the standard sugar and the tested samples could be clearly separated, When the sucrose concentration was at 0.2-0.9 mg/mL, the sucrose concentration and the peak area had good linear relation, and the corresponding R2 was 0.994 4. The instrument precision was very good with a relative standard deviation（RSD）of 2.0%. The recovery rate of the sucrose in this method was 99.7% with satisfactory value of 95%-105%. The stability of the sample was 0.9% in this test, suggesting that the sample extracted was stable and could be kept for a certain period of time.