高效液相色谱-蒸发光散射检测法测定草石蚕根茎中的水苏糖

Determination of Stachyose in Stachys Sieboldii Miq. by High Performance Liquid Chromatography with Evaporative Light Scattering Detection

应用高效液相色谱 蒸发光散射检测法建立了测定草石蚕植物根茎中水苏糖含量的新方法 ,并设计和讨论了 2种从草石蚕根茎干粉中提取水苏糖的工艺方法。采用BondapakNH2 色谱柱 (3 0 0mm× 3 .9mmi.d .) ,流动相为甲醇 水 (80∶2 0 ,V V) ,流速 1 .0mL min,柱温为室温 ;蒸发光散射检测器漂移管温度 40℃ ,载气N2 流速 4.0L min。在上述条件下测得水苏糖在 1 .0 2～ 1 2 .2 2 μg范围内线性关系良好 (r=0 9995 ) ,检出限达到 0 .1 0 μg,平均加样回收率为 91 .6%。 2种不同提取方法的测定结果 :水苏糖含量分别为 2 9.8%和 2 5 .2 % ,纯度为 63 .76%和 88.3 5 %。该方法简便快捷 ,重现性和准确度良好 ,结果准确可靠 ,为实际样品的测定提供了科学依据。

A method for the determination of stachyose in Stachys sieboldii miq. by high performance liquid chromatography (HPLC) with evaporative light scattering detection (ELSD) was established. Two different extraction crafts were designed and compared. Stachyose was successfully separated on a Bondapak NH2 column (300 mm x 3.9 mm i.d.) at ambient temperature, the mobile phase composition was a binary solvent mixture of methanol-water (80:20, V/V). The flow rate was 1.0 mL/min. The parameters of ELSD were set as follows: drift tube temperature was 40degreesC, nitrogen carrier gas flow was 4.0 L/min. This method fulfilled all the standard requirements of linearity, accuracy and precision. The linear range was 1.02similar to12.22 mug (r=0.9995). The detection limit was 0.10 mug and the average recovery was 91.6% (RSD=1.43%). The contents of stachyose by two different extraction crafts were 29.8% and 25.2%, the purity were 63.76% and 88.35%, respectively. This method was accurate, rapid, simple and reproducible, and suitable for the integrated analysis of stachyose.