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建立胰岛β-细胞膜色谱模型,以硅胶为载体,将兔胰腺β-细胞膜固定在其表面制备成胰岛β-细胞膜固定相.用lowry法测定膜蛋白含量,用电子显微镜和表面能谱分析技术,对固定相表面特性进行分析,并测定其K+,Na+-ATP酶活性.胰岛β-细胞膜色谱柱(10mm×2mm,I.D.)在(37±0.5)℃条件下,以25mmo1·L?1硫酸铵缓冲液(pH7.4)为流动相,流速为0.2mL·min?1,在不同检测波长分析了4种磺酰脲类降血糖药物格列喹酮、格列本脲、格列吡嗪和格列奇特的保留特性,用药物容量因子的对数值(logk′)表示其亲合作用强度.并与药物对小鼠正常血糖的活性强度进行了相关性分析比较.
The islet β-cell membrane chromatographic model was established, and the pancreatic β-cell membrane was immobilized on the surface of the islet β-cell membrane stationary phase by using silica gel as a carrier.The membrane protein content was determined by lowry method. The electron microscopy and surface energy spectrum analysis The surface characteristics of the stationary phase were analyzed and the activities of K +, Na + -ATPase were determined.The islet β-cell membrane column (10mm × 2mm, ID) under the condition of (37 ± 0.5) ℃ and 25mmol·L -1 ammonium sulfate The buffer solution (pH7.4) was used as the mobile phase and the flow rate was 0.2mL · min -1. Four different sulfonylurea drugs, gliquidone, glibenclamide, glipizide and The retention characteristics of gliclazide, the logarithm of the drug capacity factor (logk ’) that affinity intensity, and drug activity of normal blood glucose in mice were analyzed by correlation analysis.