HPLC法测定拉莫三嗪中的有关物质

目的：建立拉莫三嗪原料药有关物质的HPLC测定方法。方法：采用FeiniGen RedClassical Ⅱ C₁₈（250 mm×4.6 mm，5 μm）色谱柱，对拉莫三嗪的有关物质进行定量分析，以磷酸二氢钾缓冲液（取磷酸二氢钾2.7 g，加水800 mL使溶解，加三乙胺6.7 mL，用磷酸调节pH至2.0，加水至1 000 mL）为流动相A，乙腈为流动相B，梯度洗脱，流速为1.5 mL·min^-1，柱温为35℃，检测波长为210 nm（用于检测2，3-二氯苯甲酸）和270 nm（用于检测除2，3-二氯苯甲酸外的其他杂质）。用HPLC-MS鉴定氧化降解溶液中未知杂质；采用Thermo Acclaim^TM 120 C₁₈（250 mm×4.6 mm，5 μm）色谱柱，以甲醇-水（50∶50）为流动相，ESI离子源，正离子扫描方式，检测离子m/z为272。结果：主峰与各有关物质峰分离度良好，拉莫三嗪及各有关物质在质量浓度为0.3~4.0 μg·mL^-1范围内呈良好的线性关系（r²=0.998 7），拉莫三嗪的定量下限为0.2 μg·mL^-1（270 nm）、杂质Ⅰ的定量下限为0.2 μg·mL^-1（210 nm）。3批样品有关物质测定结果显示，已知杂质和未知杂质均未检出。结论：本法可为拉莫三嗪质量控制提供参考。

Objective: To establish an HPLC method for determination of related substances in lamotrigine.Methods: The test for related substances of lamotrigine was performed on the FeiniGen RedClassicalⅡC₁₈ (4.6 mm×250 mm,5 μm)column.The mobile phase A was potassium dihydrogen phosphate buffer solution(dissolve 2.7 g potassium dihydrogen phosphate in 800 mL water,add 6.7 mL triethylamine,adjust to pH 2.0 with phosphoric acid and dilute to 1 000 mL with water)and the mobile phase B was acetonitrile with gradient elution at the flow rate of 1.5 mL·min^-1;the column temperature was 35℃ and the detection wavelength was 210 nm for detecting 2,3-dichlorobenzoic acid and 270 nm for detecting impurities except 2,3-dichlorobenzoic acid.The unknown impurity in oxidative degradation solution was identified by HPLC-MS;the separation was performed on the Thermo Acclaim^TM 120 C₁₈ column(250 mm×4.6 mm,5 μm)with methanol-water(50:50)as the mobile phase.The mass spectrometer was operated in positive electrospray ionization modes with the transition m/z of 272. Results: The main peak and the peaks of the related substances were well separated.A good linearity was observed over the range of 0.3-4.0 μg·mL^-1(r²=0.998 7),the LOQ of lamotrigine was 0.2 μg·mL^-1(270 nm)and the LOQ of impurity Ⅰ was 0.2 μg·mL^-1(210 nm).The test results of three batches of samples showed that each of the impurities was not detected.Conclusion: The established method can be used for quality control of lamotrigine.

[1] 张媚,李鸣,张弦.拉莫三嗪治疗癫痫伴抑郁的临床疗效及对认知功能的影响[J].中国医药导刊,2017,19(10):1010 ZHANG M,LI M,ZHANG X.The clinical efficacy of lamotrigine in the treatment of epilepsy with depression and the influence on cognitive function[J].Chin J Med Guide,2017,19(10):1010
[2] 姚志厚.拉莫三嗪单药治疗癫痫的临床疗效和安全性观察[J].现代诊断与治疗,2018,29(6):862 YAO ZH.Clinical efficacy and safety of lamotrigine monotherapy in treating epilepsy[J].Mod Diagn Treat,2018,29(6):862
[3] ANDERSEN G,HEDERMANN G,WITTING N,et al.The antimyotonic effect of lamotrigine in non-dystrophic myotonias:a double-blind randomized study[J].Brain,2017,140(9):2295
[4] 林娜,柳洋,祁晖,等.拉莫三嗪抗抑郁作用及其机制研究[J].济宁医学院学报,2018,41(3):163 LIN N,LIU Y,QI H,et al.The effect of lamotrigine on anti-depressant of and its mechanism[J].J Jining Med Univ,2018,41(3):163
[5] 钟涛.拉莫三嗪联合丙戊酸钠治疗小儿癫痫[J].上海医药,2018,39(17):25 ZHONG T.Study on lamotrigine combined with sodium valproate in the treatment of epilepsy of children[J].Shanghai Med Pharm J,2018,39(17):25
[6] 董红梅.奥卡西平联用拉莫三嗪治疗儿童癫痫的临床对照研究[J].智慧健康,2018,4(9):49 DONG HM.Clinical control study of oxcarbazepine combined with lamotrigine in treatment of epilepsy children[J].Smart Healthcare,2018,4(9):49
[7] 田娟,李娟娟,李园,等.草酸艾司西酞普兰联合拉莫三嗪治疗儿童癫痫合并抑郁症的疗效及安全性[J].国际精神病学杂志,2017,44(6):1051 TIAN J,LI JJ,LI Y,et al.Efficacy and safety of eseitalopramoxalate combined with lamotrigine in the epilepsy children with depression[J].J Int Psychiatr,2017,44(06):1051
[8] 王凌云,侯静,张勇.左乙拉西坦联合拉莫三嗪治疗老年性癫痫患者的临床研究[J].中国实用医药,2018,13(5):3 WANG LY,HOU J,ZHANG Y.Clinical study on levetiracetam combined with lamotrigine in the treatment of elderly patients with epilepsy[J].China Pract Med,2018,13(5):3
[9] KALITA J,CHANDRA S,MISRA UK.Pregabalin and lamotrigine in central poststroke pain:a pilot study.[J].Neurol India,2017,65(3):506
[10] XU S,LIU L,CHEN Y,et al.Population pharmacokinetics of lamotrigine co-administered with valproic acid in Chinese epileptic children using nonlinear mixed effects modeling[J].Eur J Clin Pharmacol,2018,74(5):583
[11] EP 9.0.Vol Ⅲ[S].2017:2873
[12] BP 2018.Vol Ⅱ[S].2018:76
[13] USP 41-NF 36[S].2018:2333
[14] YBH01522013国家食品药品监督管理局药品标准[S].2013 YBH01522013 The State Food and Drug Administration Drug Standard[S].2013
[15] YBH04392010国家食品药品监督管理局药品标准[S].2010 YBH04392010 The State Food and Drug Administration Drug Standard[S].2010
[16] 中华人民共和国药典2015年版.四部[S].2015:374 ChP 2015.Vol Ⅳ[S].2015:374
[17] 谢沐风,罗霞萍,陈亚美.如何建立HPLC法测定有关物质的方法[J].中国药品标准,2002,3(6):6 XIE MF,LUO XP,CHEN YM.How to build the method of the related substance analysis by HPLC[J].Drug Stand China,2002,3(6):6