芪蛭降糖胶囊中毛蕊异黄酮葡萄糖苷薄层鉴别及含量测定

TLC identification and content determination of calycosin-7-O-β-D-glucoside in Qizhi jiangtang capsule

目的建立中药复方芪蛭降糖胶囊中毛蕊异黄酮葡萄糖苷的薄层鉴别和含量测定方法。方法采用薄层色谱法(TLC)对组方中毛蕊异黄酮葡萄糖苷进行定性鉴别,采用70%乙醇溶液提取、D101大孔树脂富集,以硅胶GF_(254)板为吸附剂,三氯甲烷-甲醇-水为展开系统,紫外显色;利用高效液相色谱法(HPLC)对其进行含量测定,色谱柱为Agilent SB-C_(18)色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%磷酸水,检测波长260 nm,流速1 ml/min,柱温30℃,进样量10μl。结果薄层色谱斑点分离效果较好(Rf=0.44),斑点显色清晰且无干扰、重复性好;高效液相色谱毛蕊异黄酮葡萄糖苷峰分离良好,线性方程为Y=1035.8X+509.89(r=0.999 6,0.716~3.580μg),线性关系良好。结论该方法可用于芪蛭降糖胶囊中毛蕊异黄酮葡萄糖苷的定性鉴别和含量测定。

Objective To set up the methods of TLC identification and content determination of calycosin-7-O-β-D-glucoside in Qizhi jiangtang capsule. Methods The thin layer chromatography（ TLC） was used to give the qualitative identification of calycosin-7-O-β-D-glucoside. 70% ethanol solution was used for the extraction,the macroporous resin D101 for purification,the silica gel GF_（245） plate as adsorbent,the trichloromethane-metnanol-water for the spreading system and ultraviolet spectrophotometry method for coloration. The high performance liquid chromatography（ HPLC） was adopted for the content determination,with Agilent SB-C_（18） column,250 mm × 4. 6 mm,5 μm; acetonitrile -0. 1% phosphoric acid as mobile phase,at 260 nm in wavelength,1 ml/min as flow velocity and 30 ℃ at temperature. The sample size was 10 μl. Results The separation effect of TLC spots was good（ Rf = 0. 44）. They were clear,no interference and good in repeatability. HPLC showed the good peak separation of calycosin-7-O-β-D-glucoside. The linear equation was Y = 1035. 8 X ＋ 509. 89（ r = 0. 999 6,0. 716 ~ 3. 580 μg）,indicating the good linear relationship. Conclusion This method is applicable for the qualitative identification and content determination of calycosin-7-O-β-D-glucoside in Qizhi jiangtang capsule.