反相高效液相色谱法测定小鼠血浆中醋酸洗必泰浓度

Determination of Chlorhexidine Acetate in Mouse Serum by RP-HPLC

目的 :建立小鼠血浆中醋酸洗必泰浓度的反相高效液相色谱测定法。方法 :以十二烷基磺酸钠为反离子 ,安定为内标 ,采用离子对萃取法对血样进行提取 ;μ-BondapakC18 柱 (3 9mm×300mm ,10μm)为分析柱 ,0 02mol/L磷酸二氢钾缓冲液 (pH=3)∶甲醇(44:56 ,V/V)为流动相 ,检测波长为257nm。结果 :醋酸洗必泰在0 5～10μg/ml血药浓度范围内 ,浓度与峰面积比值间线性关系良好 ,r=0 9997 ;3种血浆浓度 (1、4、8μg/ml)平均回收率为104 67 % ,日内、日间精密度分别为1 5 %、0 37 %、0 26 %及2 45 %、0 46 %、1 27 % ;血浆中药物的最低检测浓度为0 4μg/ml。结论 :该方法灵敏、准确 ,适用于临床药代动力学的研究。

OBJECTIVE:To develop a RP-HPLC method for determining the concentration of chlorhexidine(Ⅰ) in mouse serum METHODS:Ⅰ and the internal standard,diazepam,were extracted to ethyl acetate with sodium laurysulfonate as ion-pair Chromatographic separation was achieved on μ-Bondapak C18 column(3 9mm×300mm,10μm)equilibrated with 0 02mol/L potassium dihydrogen phosphate buffer solution (pH=3)∶methanol(44∶56,V/V),with UV detection at 257nm RESULTS:The calibrotion curve of Ⅰ was linear in the range of 0 5～10μg/ml,r=0 9 997 The avarage recovery was 104 67%(n=6) The within-day and between-day RSDs in serum concentrations of 1?4?8μg/ml were 1 5%?0 37%?0 26% and 2 45%?0 46%?1 27% respectively The minimum detectable concentration was 0 4μg/ml CONCLUSION:The developed method was sensitive,accurate and suitable for the determination of Ⅰ in mouse serum