RP-HPLC法同时测定家兔体内氨苯砜、咖啡因和美托洛尔的含量

目的: 建立同时测定家兔体内CYP450 3种探针药物氨苯砜、咖啡因、美托洛尔含量的方法。 方法: 用反相高效液相色谱法测定,选用Diamonsil C₁₈色谱柱(200 mm×4.6 mm,5 μm);流动相为甲醇-水-冰醋酸-三乙胺(20∶ 80∶ 0.06∶ 0.1);检测波长:氨苯砜为295 nm,咖啡因为275 nm,美托洛尔为225 nm;流速为1.5 mL·min^-1;柱温为35 ℃。 结果: 在上述色谱条件下,3种探针药物可以很好地分离和测定,氨苯砜、咖啡因和美托洛尔的线性范围分别为0.050~62.5 μg·mL^-1(r=0.9999),0.10~40.0 μg·mL^-1 (r=0.9996),0.10~25.0 μg·mL^-1 (r=0.9999);方法回收率分别为:氨苯砜85.6%~101.3%,咖啡因96.1%~112.8%,美托洛尔93.3%~102.7%。 结论: 该方法简便快速,结果准确,可用于同时测定家兔体内氨苯砜、咖啡因和美托洛尔的含量,为研究受试药物对CYP450各亚酶的调控作用及相关研究提供了分析测定方法。

Objective: To establish a method for simultaneously determining the contents of three probe drugs of CYP450 in rabbit plasma,including dapsone,caffeine and metoprolol. Methods: The three drugs were determined by RP-HPLC,and the chromatography conditions were as follows:Diamonsil C₁₈ column (200 mm×4.6 mm,5 μm),mobile phase consisting of methanol,water,glacial acetic acid and triethylamine (20∶ 80∶ 0.06∶ 0.1),flow rate 1.5 mL·min^-1,the detection wavelength for dapsone at 295 nm,caffeine at 275 nm and metoprolol at 225 nm,column temperature 35 ℃. Results: The three drugs were seperated well under the above HPLC condition.There was good linear relationship for dapsone in the ranges of 0.050-62.5 μg·mL^-1(r=0.9999),caffeine 0.10-40.0 μg·mL^-1 (r=0.9996),and metoprolol 0.10-25.0 μg·mL^-1 (r=0.9999).The recoveries of dapsone,caffeine and metoprolol were 85.6%-101.3%,96.1%-112.8%,and 93.3%-102.7,respectively. Conclusion: The method is simple and accurate,and suitable for simultanously determining plasma concentrations of dapsone,caffeine and metoprolol in rabbit,and can be used for evaluating effects of drugs on CYP450 subtype enzymes activities and related study.