摘要
目的:建立高效液相色谱-迎头分析法(HPLC-FA)研究格列本脲与人血清白蛋白(HSA)的结合作用。方法:采用HPLC-FA法,色谱柱为 Pinkerton GFFⅡ-S5-80(150 mm×4.6 mm,5 μm),流动相为67 mmol·L^(-1)磷酸盐等渗缓冲液(pH 7.4,I=0.17 mol·L^(-1)),流速0.2 mL·min^(-1),检测波长230 nm,进样量900μL,柱温37℃。结果:应用非线性回归参数估算求得 HSA 分子上两类结合位点结合的格列本脲分子数及相应的结合平衡常数:n_1,=1.5和 K_1=1.1(μmol·L^(-1))^(-1);n_2=3.8和 K_2=0.012(μmol·L^(-1))^(-1)。结论:HPLC-FA 法操作简便,适用于研究格列本脲与 HSA 的结合作用。本实验选定的进样体积为900μL。
Objective:To develop HPLC - FA method for study of protein binding of glibenclamide. Methods: The separation is performed using Pinkerton GFF Ⅱ - S5 - 80 internal - surface reversed - phase silica support ( 150 mm ×4. 6 mm,5 μm)at pH 7.4 in a 67 mmol · L^-1 isotonic sodium phosphate buffer at 37 ℃. Other conditions included flow rate 0.2 mL · min^ - 1, UV detection at wavelength 230 nm and injection volume 900 μL. Resuits:Nonlinear regression parameter estimation is applicable to the association constants measurement of glibenclamide to both primary and secondary sites, which are 1.1 ( μmol · L^-1 )^ -1 and 1.5 for K1 and n1 , respectively,and 0. 012( μ mol · L^-1 )^ -1 and 3.8 for K2 and n2 ,respectively. Conclusions:The method is proved to be suit- able for investigation of protein binding of glibenclamide. The injection volume is 900 μL.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2005年第10期1176-1178,共3页
Chinese Journal of Pharmaceutical Analysis