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建立了用高效液相色谱检测红细胞混悬液中盐酸普萘洛尔的方法并用于红细胞膜β2受体-配体结合实验研究。通过1500 r/m in离心5 m in的方法将试样中游离普萘洛尔和含受体-普萘洛尔复合物的红细胞分离,采用HypersilBDS?C18柱(150 mm×4.6 mm,5μm)为分析色谱柱;流动相为20 mmol/LKH2PO4溶液(其中含0.25%三乙胺,pH 5.1)?甲醇(65∶35,V/V),流速为1.0 mL/m in;276 nm波长处检测;柱温30℃;进样量20μL。普萘洛尔在5~150 ng/mL范围内线性关系良好,相关系数为0.9998。样品测定的重复性相对标准偏差为1.05%;检出限为1.5 ng/mL。样品测定结果表明普萘洛尔的特异性结合量随其加入量的增加而增大,然后达到饱和。该方法可用于受体配体结合实验的研究。
A method for the determination of propranolol hydrochloride in erythrocyte suspension by high performance liquid chromatography (HPLC) was established and used to study the β2 receptor-ligand binding of erythrocyte membrane. Samples were separated from free propranolol and receptor-propranolol complex-containing red blood cells by centrifugation at 1500 rpm for 5 minutes, using a Hypersil BDS® C18 column (150 mm × 4.6 mm, 5 μm) The mobile phase consisted of 20 mmol / L KH2PO4 solution (containing 0.25% triethylamine, pH 5.1) and methanol (65:35, V / V) at a flow rate of 1.0 mL / ; Column temperature 30 ℃; injection volume 20μL. Propranolol had a good linearity in the range of 5-150 ng / mL with a correlation coefficient of 0.9998. The relative standard deviation (RSD) of the sample was 1.05%. The detection limit was 1.5 ng / mL. The results of sample measurement showed that the specific binding of propranolol increased with the addition of propranolol and then reached saturation. This method can be used for the study of receptor ligand binding experiments.