浊点萃取-高效液相色谱法同时测定槲寄生中3种黄酮的含量

Determination of three flavonones in Viscum coloratum(Komar.) Nakai by HPLC-UV with cloud-point extraction

目的采用浊点萃取法富集槲寄生中的异鼠李素-3-O-β-D-葡萄糖苷、鼠李秦素-3-O-β-D-葡萄糖苷、鼠李秦素-3-O-β-D-(6″-β-羟基-β-甲基戊二酸半酯)葡萄糖苷,并使用高效液相色谱法测定其含量。方法采用聚乙二醇辛基苯基醚(Triton X-114)作为萃取剂,色谱柱为Synergi C18柱(250 mm×4.6 mm,4μm),流动相为乙腈(A)和体积分数0.5%乙酸溶液(B)梯度洗脱,流速为1.0 mL.min-1,柱温为30℃,检测波长为358 nm。结果测得异鼠李素-3-O-β-D-葡萄糖苷、鼠李秦素-3-O-β-D-葡萄糖苷、鼠李秦素-3-O-β-D-(6″-β-羟基-β-甲基戊二酸半酯)葡萄糖苷质量浓度分别在2.093～41.86、1.200～24.00、0.861 0～17.22 mg.L-1内与峰面积呈良好线性关系;平均回收率分别为93.2%、95.6%、94.5%。结论该方法准确、灵敏,可作为槲寄生药材质量控制的有效方法。

Objective To develop a method based＇on cloud-point extraction （CPE）for the determination of isorhanmetin-3-O-β-D-glucoside, rhamnazin-3-O-β-D-glucoside, rhamnazin-3-O-β-D-（ 6＂-β-hydroxy-fl-methylglutaryl）-glucoside in the Viscum coloratum（ Komar. ）Nakai by HPLC-UV. Methods The non-ionic surfactant Triton X- 114 was chosen as the extract solvent and various experimental conditions were investigated to optimize the extraction process. Under optimum conditions, the extracts were analyzed by reversephase column. Acetonitrile and 0. 5 % acetic acid were used as the mobile phase under gradient conditions. The flow rate was 1.0 mL· min^-1 and the UV detection wavelength was set at 358 nm. Results All calibration curves showed good linearity （ r 〉 0. 999） within test ranges. The recoveries, measured at three concentration levels, varied from 93.2% to 95.6%. Conclusions The developed and validated methods are simple, accurate and can be selected as the techniques for separation and quantification of the three flavonones in the Viscum coloratum（ Komar. ） Nakai.