反相高效液相色谱法测定(E)-3-(4-((3,5,6-三甲基吡嗪-2-基)甲氧基)-3-甲氧基苯基)丙烯酸在SD大鼠血浆中的浓度

Determination of(E)-3-(3-methoxy-4-((3,5,6-trimethylpyrazin-2-yl) methoxy)phenyl)acrylate in rat plasma by RP-HPLC

目的建立测定SD大鼠血浆中(E)-3-(4-((3,5,6-三甲基吡嗪-2-基)甲氧基)-3-甲氧基苯基)丙烯酸浓度的反相高效液相色谱(LC-UV)方法,并用于其在SD大鼠体内的药动学研究。方法采用Diamonsil-C18(2)柱(100 mm×4.6 mm,5μm),以乙腈∶水(含5 mM的醋酸铵,用醋酸调节pH至4.0)=30∶70为流动相,测定70只SD大鼠单剂量尾静脉给药15mg/kg后不同时刻血浆中药物的浓度,并由此计算其在SD大鼠体内的药动学参数。结果 SD大鼠血浆中(E)-3-(4-((3,5,6-三甲基吡嗪-2-基)甲氧基)-3-甲氧基苯基)丙烯酸测定的线性范围为0.020 16～10.08μg/ml,提取回收率大于95%,批内批间精密度均小于15%。大鼠尾静脉注射给药后,血浆中峰浓度(cmax)为35.60μg/ml,半衰期(t1/2)为3.36 h,0～12 h药时曲线下面积(AUC0～12)为17.12μg.h/ml,0～∞药时曲线下面积(AUC0～∞)为17.72μg h/ml。结论本法方便快捷,可用于(E)-3-(4-((3,5,6-三甲基吡嗪-2-基)甲氧基)-3-甲氧基苯基)丙烯酸的临床前药动学研究。

Objective To establish a method for determination of （ E ） -3- （ 3-methoxy-4- （ （ 3,5,6-trimethylpyrazin-2-yl ） me- thoxy） phenyl）acrylate in plasma of SD rats by RP-HPLC. Methods The chromatographic separation was performed on a Diamonsil- C18 column（ 100 mm× 4.6 mm,5 μm） with the mobile phase consisting of acetonitrile/5 mM ammonium acetate buffer（ pH 4.0） （30/ 70, v/v ）. The single intravenous injection of （E）-3-（ 3-methoxy-4-（ （3,5,6-trimethylpyrazin-2-yl） methoxy）phenyl）acrylate was given to 70 SD rats to study the pharmaeokineties. Results The intra-run and inter-run RSD were less than 15%. The pharmacokinetics parameters were as follows： Cmox was 35.60 pog/ml , t1/2 was 3.36 h, AUC0-12 was 17. 12 μg h/ml, AUC0-∞ was 17.72μg h/ml. Conclusion The method was simple and accurate, which could be used to separation and determination of （E）-3-（3-methoxy-4-（ （3, 5,6-trimethylpyrazin-2-yl） methoxy） phenyl） acrylate.