沙丁胺醇与红细胞膜表面β_2-肾上腺素受体结合量的高效液相色谱法测定  

Determination of Salbutamol Binding Capacity with β_2-Adrenergic Receptor in Avian Erythrocyte Membrane by HPLC

作　　者：蒋心惠[1] 杨俊卿 周岐新[1] 

机构地区：[1]重庆医科大学药学院,重庆400016

出　　处：《分析测试学报》2010年第8期837-840,845,共5页Journal of Instrumental Analysis

基　　金：国家自然科学基金资助项目(30572353)

摘　　要：建立了同时检测红细胞混悬液中沙丁胺醇（SA）与普萘洛尔（PR）含量的高效液相色谱法，并用于红细胞膜殷受体一配体结合实验研究。药物与红细胞悬液保温反应1h后，以1500r／min离心5min，将试样中游离SA和PR与含大分子的受体一药物复合物的红细胞分离，以HypersilC18柱为分析柱，10mmol／L磷酸二氢钾溶液（含0．2％三乙胺，pH5．1）-甲醇（体积比17：83）为流动相；于激发波长284nm，发射波长323nm处检测。SA在4．0～48．0μg／L范围内线性关系良好，相关系数为0．9989；PR在2．0—12．0mg／L范围内线性关系良好，相关系数为0．9998。SA与PR的RSD分别为0．85％、0．79％；检出限分别为0．18、0．30μg/L。测定结果表明，sA的非特异结合和特异结合量均随着sA加入量的增加而增大。该方法可以用于药物和受体相互作用的研究。An effective high performance liquid chromatographic method was developed for the simultaneous determination of salbutamol （SA） and propranolol （PR） in erythrocyte suspensions in order to investigate SA binding to β2 receptor. Free SA and PR were separated from erythrocyte through 5 minutes centrifugation with a speed of 1 500 r/min after SA and PR reacting with membrane receptor existing on the surface of erythrocytes for 1 hour. The analysis of SA and PR was performed on a hypersil C18 column with 10 mmol/L potassium dihydrogen phosphate solution containing 0. 2% triethylamine（pH 5.1） -methanol（17 ： 83, by volume） as mobile phase. The fluorescence detector was selected with 284 nm as excitation wavelength and 323 nm as emission wavelength. The calibration curves were linear in the range of 4.0 - 48.0 ixg/L for SA with a correlation coefficient of 0. 998 9 and 2. 0 - 12. 0 mg/L for PR with a correlation coefficient of 0. 999 8. The relative standard devia- tions of SA and PR were 0. 85% and 0. 79% , respectively, and the limits of detection（LODs） were 0. 18 μg/L and 0. 30 μg/L, respectively. The results indicated that the non-specific binding and specific binding capacities of salbutamol to β2 receptor increased with the increase of SA concentration. The method was suitable for the investigation of the interaction between drugs and receptors.

关 键 词：沙丁胺醇 普萘洛尔 红细胞膜 β2-肾上腺素受体 高效液相色谱 

分 类 号：O657.72[理学—分析化学] TQ460.72[理学—化学]