高效液相色谱法研究沙丁胺醇与红细胞膜表面β_2肾上腺素受体结合  

Determination of Salbutamol and Its Combination with β_2-Adrenergic Receptor from Avian Erythrocyte Membrane by HPLC

作　　者：蒋心惠[1] 滕永真[2] 周岐新[1] 

机构地区：[1]重庆医科大学药学院,重庆市渝中区医学院路1号400016 [2]重庆医科大学生命科学院,重庆市渝中区医学院路1号400016

出　　处：《光谱实验室》2012年第4期1960-1963,共4页Chinese Journal of Spectroscopy Laboratory

基　　金：国家自然科学基金(30572353)

摘　　要：建立了同时检测红细胞混悬液中沙丁胺醇(Salbutamol,SA)与特布他林(Terbutaline,TE)的高效液相色谱法并用于沙丁胺醇与红细胞膜β_2受体-配体结合实验研究。通过1500r/min离心5min的方法将试样中游离SA和TE与含大分子的受体-药物复合物的红细胞分离,以10mmol/L磷酸二氢钾溶液(其中含0.2%三乙胺,用0.5mol/L磷酸调pH 5.1)-甲醇(90:10,V/V)为流动相;流速为1.0mL/min;检测波长为276nm;柱温30℃;进样量20μL。SA在2.0—20.0ng/mL范围内线性关系良好,相关系数为0.9965;TE在0.20—20μg/mL范围内线性关系良好,相关系数为0.9971。样品测定SA和TE的重复性RSD分别为1.26%和0.94%;检出限分别为0.5ng/mL和1.0ng/mL。测定结果显示,SA的非特异结合量和特异结合量均随SA加入量的增加而增大。TE和SA与β_2肾上腺素受体的结合位点不同。An effective HPLC method with hypersil C18column,10 mmol/L potassium dihydrogen phosphate solution（contains 0.2%triethylamine,adjusted pH to 5.1 with 0.5 mol/L phosphate）-methanol（90：10,V/V） as mobilc phase and detection wavelength at UV 276nm has been developed for simultaneous determination of salbutamol（SA） and terbutaline（TE） in erythrocyte suspensions in order to investigate SA binding toβ2 receptor.The flow rate was 1.0mL/min and the column tempreture was 30 C.Free SA and TE were separated from the membrane receptor-binding complex through 1500r/min centrifugal for 5min.The linear ranges and correlation coefficients were 2.0-20.0ng/mL and 0.9965 for SA as well as 0.20-20μg/mL and 0.9971 for TE.The RSDs were 1.26%for SA and 0.94%for TE.The LODs of SA and TE were 0.5ng/mL and 1.0ng/mL,respectively.The results show that the non-specific binding and specific binding amounts of salbutamol toβ2 receptor increase with the increasing of SA amount used,and the sites of TE or SA binding withβ2-adrenergic receptor are different.

关 键 词：沙丁胺醇 特布他林 红细胞膜 Β2肾上腺素受体 高效液相色谱法 

分 类 号：O657.7[理学—分析化学]