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采用HPLC法测定15个不同产地甜叶菊叶中莱鲍迪苷A的含量,筛选适合甜叶菊叶水提物中富集总糖的吸附树脂,建立洗脱条件。HPLC条件为:Agilent-C18(4.6x250 mm,5μm)色谱柱;流动相为75%甲醇-水(V/V)等度洗脱,流速1 m L/min;柱温30℃;测定波长210 nm;进样量20μL。结果表明,各地区甜叶菊叶中莱鲍迪苷A的含量范围在0.25%~17.50%之间,差异较大;浙江绍兴、浙江衢州和新疆阿拉尔产甜叶菊叶中莱苞迪苷A含量较高,超过13.0%。不同型号树脂对莱鲍迪苷A的吸附量存在差异,LSA-10,D101,S-8,AB-8,ADS-21和HPD450较适合从水提液中富集总糖,且莱鲍迪苷A的相对含量较高。采用D101树脂吸附总糖的较优洗脱条件为:首先用3%Na OH水溶液洗脱除杂,接着水洗至中性,最后用90%乙醇-水洗脱收集总糖。
The contents of rebaudioside A in 15 leaves of Stevia rebaudiana L. were determined by HPLC, and the adsorption resin suitable for total sugar in the water extract of Stevia rebaudiana was screened. The elution conditions were established. The HPLC conditions were: Agilent-C18 column (4.6x250 mm, 5μm); mobile phase consisted of isocratic elution of 75% methanol / water with a flow rate of 1 m L / min; nm; injection volume 20μL. The results showed that the contents of rebaudioside A in stevia rebaudiana leaves ranged from 0.25% to 17.50% in different regions, and the differences were significant. The content of rebaudioside A in stevia rebaudiana leaves from Shaoxing, Zhejiang Province and Alar of Xinjiang, Higher, more than 13.0%. The adsorption amount of rebaudioside A in different types of resin is different. LSA-10, D101, S-8, AB-8, ADS-21 and HPD450 are more suitable for the enrichment of total sugar from aqueous extract, The relative content of glycoside A is higher. The optimum conditions for the total sugar adsorption on D101 resin were as follows: firstly, the impurities were eluted with 3% NaOH aqueous solution, then washed with water until neutral, and finally with 90% ethanol-water.