高效液相色谱法测定安立生坦中对映异构体

目的: 建立手性高效液相色谱方法,用于测定安立生坦中对映异构体。方法: 采用对照品法进行定性定量,使用Chiralpak AD-H(4.6mm×250 mm,5 μm)色谱柱,流动相为正己烷-乙醇(95:5),流速1.0 mL·min^-1,检测波长263 nm。结果: 安立生坦与其对映异构体之间有效分离,分离度大于8.0;质量浓度在1.04~5.18 μg·mL^-1范围内线性关系良好;低、中、高浓度平均加样回收率(n=3)分别为106.6%、103.4%、103.7%,RSD分别为3.0%、1.4%、2.2%;样品溶液在24 h内基本稳定。经检测,6批安立生坦中对映异构体含量分别为0.15%、0.13%、0.09%、未检出、0.12%和0.16%。结论: 本方法经方法学验证,可用于安立生坦中对映异构体的测定。

Objective: To develop an HPLC method for the determination of enantiomer in ambrisentan. Methods: Reference substance method was used for the qualitative and quantitative determination.The separation was performed on a Chiralpak AD-H column (4.6 mm×250 mm, 5 μm) with the mobile phase of hexane-ethanol (95: 5) at a flow rate of 1.0 mL·min^-1, and the detective wavelength was set at 263 nm. Results: Ambrisentan and enantiomer were effectively separated under the above conditions with the resolution higher than 8.0;the calibration curves were linear in the concentration range of 1.04-5.18 μg·mL^-1;the recoveries (n=3) were 106.6%, 103.4%, and 103.7% respectively at low, medium and high concentrations, with the RSD of 3.0%, 1.4%, and 2.2%, respectively;the test solution was basically stable within 24 h.The enantiomer contents in six batches of ambrisentan were 0.15%, 0.13%, 0.09%, not detected, 0.12% and 0.16%respectively. Conclusion: The method is suitable for determination of enantiomer in ambrisentan by methodology validation.

[1] HROMETZ SL,SHIELDS KM.Role of ambrisentan in the management of pulmonary hypertension[J].Ann Pharmacother,2008,42(11):1653
[2] CHENG JW.Ambrisentan for the management of pulmonary arterial hypertension [J].Clin Ther,2008,30(5):825
[3] Assessment Report for Volibris,EMEA123999.2008
[4] HARBESON S.Selective Endothelin Type-A Antagonists:WO 2008097468 A2[P].2008-08-14
[5] GU JM(谷建敏),DU YM(杜玉民),WANG W(王伟),et al.Determination of dissociation constants of darusentan and ambrisentan by capillary electrophoresis(毛细管电泳法测定达卢生坦与安贝生坦的解离常数)[J].Chin J Pharm(中国医药工业杂志),2012,43(6):468
[6] SONG J(宋杰),YU ZX(余再新).Endothelin receptor antagonists for pulmonary arterial hypertension(内皮素受体拮抗剂在肺动脉高压治疗中的应用)[J].Clin Med J(临床药物治疗杂志),2013,11(6):5
[7] RIECHERS H,ALBRECHT HP,AMBERG W,et al.Discovery and optimization of a novel class of orally active nonpeptidic endothelin-A receptor antagonists[J].J Med Chem,1996,39(11):2123
[8] ZHANG L(张玲),YANG HL(杨华良),WANG JM(王家明).Determination of the six residual organic solvents in ambrisentan by GC(GC测定安立生坦原料药中6种有机溶剂残留)[J].Chin J Mod Appl Pharm(中国现代应用药学),2014,31(10):1238
[9] ZHANG DD(张丹丹),YAN CQ(闫翠勤),QIAO HM(乔红梅).Enantiomeric separation of 2-hydroxy-3-methoxy-3,3-diphenylpropionic acid by high performance liquid chromatography(高效液相色谱法分离2-羟基-3-甲氧基-3,3-二苯基丙酸对映体)[J].Appl Chem Ind(应用化工),2013,42(8):1544
[10] LI XG(李小刚).Study on the synthesis of ambrisentan(安立生坦的合成研究)[J].Appl Chem Ind(应用化工),2014,43(9):1675
[11] ZHOU FG(周付刚),GU JM(谷建敏),SU XJ(苏信杰),et al.Synthesis of ambrisentan(安贝生坦的合成)[J].Chin J Pharm(中国医药工业杂志),2010,41(1):60
[12] MATIORAM G,CHRISTIAN J,ALEXANDRE M,et al.Process for Producing Ambrisentan:WO 2010091877 A2[P].2010-8-19