HPLC/DAD/MS法测定氯雷他定及其微量杂质

目的: 建立高效液相色谱/二极管阵列检测器/质谱法(HPLC/DAD/MS)法测定氯雷他定及其2种合成过程中微量杂质地洛他定和N-甲基地洛他定,并对另一未知杂质结构进行鉴定。 方法: 采用RP-HPLC法,色谱柱:Alltima C₁₈柱(150 mm×4.6 mm,5 μm);流动相:乙腈-2%醋酸水溶液梯度洗脱,梯度条件如下:0 min,10∶ 90;5 min,10∶ 90;5.10 min,30∶ 70;15.00 min,30∶ 70;15.10 min,50∶ 50;40.00 min,50∶ 50;流速:1.0 mL·min^-1;柱温:35 ℃;进样量:10 μL;检测波长:276 nm;离子源:ESI,扫描模式:正离子扫描。未知杂质的分离制备采用RP-HPLC法,色谱柱:Alltima C₁₈半制备柱(250 mm×10 mm,10 μm);流动相: 乙腈-2%冰醋酸水溶液70∶ 30等度洗脱,流速3.8 mL·min^-1;检测波长296 nm。采用600 M核磁共振仪对未知杂质X进行¹H-NMR的测定。 结果: 氯雷他定及其2种杂质分离良好。氯雷他定、地洛他定、N-甲基地洛他定分别在0.0102~10.2 μg、0.00508~5.08 μg、0.0053~5.32 μg范围内线性关系良好(r=0.9996~0.9999);最低检测限分别为0.31,0.15,0.16 ng;定量限分别为1.02,0.51,0.53 ng。推测了该未知杂质X的化学结构为8-氯-11(1-羧酸乙酯基-4-哌啶基)-11H-苯并 芳庚并 吡啶,为氯雷他定的异构体。 结论: 该方法简便准确,可靠,重现性好,方法可行,可应用于原料药和制剂的质量研究和质量控制。

Objective: To establish a method of high performance liquid chromatography-diode array detection-mass spectrometry(HPLC/DAD/MS)to determine loratadine and its impurities which produced during the synthesis progress and identify the structure of an unknown impurity. Methods: Two HPLC methods were established.One method was achieved on a C₁₈ column(150 mm×4.6 mm,5 μm)utilizing a mobile phase of acetonitrile-2% acetic acid solution(0 min,10∶ 90;5 min,10∶ 90;5.10 min,30∶ 70;15.00 min,30∶ 70;15.10 min,50∶ 50;40.00 min,50∶ 50)at a flow rate of 1.0 mL·min^-1 with a diode array detector set at 276 nm.The other method was achieved on a C₁₈ semipreparative column(250 mm×10 mm,10 μm)utilizing a mobile phase of acetonitrile-2% acetic acid solution(70∶ 30)at a flow rate of 3.8 mL·min^-1 with a diode array detector set at 296 nm for the unknown impurity separation and preparation.The chemical structure of the unknown impurity was elucidated by NMR spectra of ¹H-NMR. Results: Loratadine and its impurities could be separated with a good resolution.Loratadine,desloratadine and N-methyldesloratadine were liner over the range of 0.0102-10.2 μg,0.00508-5.08 μg,0.00532-5.32 μg(r=0.9996-0.9999);The values of limit of detection were 0.31,0.15,0.16 ng,and values of limit of quantitation were 1.02,0.51,0.53 ng,respectively.The chemical structure of unknown impurity X had been identified as 8-chloro-11(1-ethoxycarbony-4-piperidinyl)-11H-benzo cyclohepta pyridine,which is one of isomers of loratadine. Conclusion: The method appeared to be simple,accurate,reliable,and reproducible,and can be applied to quality research and quality control of loratadine and its preparations.