草石蚕水苏糖检测技术的研究

Determination and Characteristic of Stachyose in Stachys SieboldiiMiq

建立了我国传统植物草石蚕块茎中水苏糖的定性分析和定量检测方法。采用薄层层析(TLC)法定性分析,选择乙酸乙酯/甲醇/水/冰醋酸(3/2/1/2)为展开剂,采用二苯胺-苯胺-磷酸为显色剂。该条件下水苏糖Rf值为0.47,显色点为棕黄色,斑点清晰,与其他糖分分离良好。用高效液相色谱-示差折光检测(HPLC-RID)法定量检测,采用Kromasil NH_2柱,流动相为乙腈/水(55/45,v/v),柱温为30℃,流速为1.0 mL/min,进样体积10μL,该色谱条件下,水苏糖峰出峰对称性好,时间较短(9.5 min),水苏糖浓度在1.0～8.0 mg/mL范围内线性关系良好(R^2=0.9996),回收率在96.0%～102.5%之间。在生产应用中,可以根据实际采用其中一种或者两种方法结合,以满足检测需要。

The methods of qualitative and quantitative determinations on stachyose extracted from the Stachys Sieboldii Miq. have been established. In the method of thin-layer chromatography （TLC） ,ethyl acetate/methanol/water/acetlc acid （3/2/1/2） are selected as the developing agent, diphenylamine-aniline-phosphate as the reagent. Under this condition ; Rf value of stachyose is 0. 47, with yellow spot, separated from other sugars clearly. High performance liquid chromatography （HPLC） is used with the refractive index detector （RID）, Kromasil NH2 column, mobile phase of acetonitrile/water （55/45, v/v） ,column temperature 30℃ and the flow rate 1.0 mL/min. The results show that the stachyose peak have great symmetry, short time （9.5 min）, good linear relationship （R2 = 0. 9996） with concentration of 1.0 - 8.0 mg/mL and recoveries of 96.0% - 102. 5 %. In production applications, one or combine two methods can be chosen for the testing requirements.