目的：建立无催化剂作用下衍生化GC-MS 法测定利伐沙班原料药中基因毒杂质5-氯-2-酰氯噻吩。方法：采用气相色谱-质谱联用法,色谱柱为Agilent HP-INNOWAX 毛细管柱（30 m×0.32 mm×0.25μm）,柱温为程序升温（起始温度110 ℃,保持2 min,10 ℃·min-1 升温至230 ℃,保持6 min）,进样口温度230 ℃,分流比5∶1,氦气流速1.5 mL·min-1,直接进样体积1 μL;离子源为EI,电子能量70 eV,离子源温度230 ℃,四极杆温度150 ℃,扫描方式及碎片离子分别为SIM 及m/z 145、m/z 176。结果：基因毒杂质5-氯-2-酰氯噻吩质量浓度在8.21~101.47 ng·mL-1 范围内与峰面积积分值呈良好的线性关系,相关系数r=0.998 1,平均加样回收率（n=9）为99.2%,精密度试验RSD（n=6）为2.5%,且衍生化产物在常温下8 h内稳定。6批样品的基因毒杂质5-氯-2-酰氯噻吩含量结果分别为6.48、6.94、1.83、46.70、49.93、46.35ng·mL-1。结论：本方法专属性强,准确度、精密度高,耐受性、灵敏度好,可用于利伐沙班原料药中基因毒杂质5-氯-2-酰氯噻吩的检测和质量控制。
Objective: To establish a method for the determination of genotoxic impurity（5-chlorothiophene-2-carbonyl chloride）in rivaroxaban by GC-MS with catalyst-free derivatization. Methods: GC-MS method was adopted. The determination was performed on capillary column of Agilent HP-INNOWAX（30 m×0.32 mm,0.25 μm）;the column temperature was programmed to start at 110 ℃,maintained for 2 min,and then rose to 230 ℃ at a rate of 10 ℃·min-1,maintained for 6 min;the inlet temperature was 230 ℃,helium （1.5 mL·min-1）was used as the carrying gas;the split ratio was 5∶1;the injection volume was 1 μL. The detector was MS;the ion source was EI;the electron energy was 70 eV;the temperature of the ion source was 230 ℃ ; the temperature of the fourth pole was 150 ℃ ;scanning mode and fragment ions were SIM,m/z 145,m/z 176, respectively. Results: The genotoxic impurity（5-chlorothiophene-2-carbonyl chloride）showed good linear relationships within the ranges of 8.21-101.47 ng·mL-1（r=0.998 1）. The average recovery was 99.2%（n=9）. RSD（n=6）of precision test was 2.5% and the derivative product was stable within 8 hours at room temperature. The contents of 5-chloro-2-acylthiphene in six samples were 6.48,6.94,1.83,46.70,49.93,46.35 ng·mL-1, respectively. Conclusion: This method has high specificity,accuracy,precision and tolerance sensitivity,which can be used for the test and quality control of the genotoxic impurity（5-chlorothiophene-2-carbonyl chloride）of rivaroxaban.
[1] 江凯,张惠斌,周金培,等.Xa 因子抑制剂的研究进展[J].中国新药杂志,2012,21(20):2080
JIANG K,ZHANG HB,ZHOU JP,et al.Research progress of XA factor inhibitors[J].Chin J New Drugs,2012,21(20):2080
[2] 吴丹,王静,王艳,等.噁唑烷酮类化合物的合成及其凝血因子Xa 抑制活性研究[J].中国药物化学杂志,2019,29(6):426
WU D,WANG J,WANG Y,et al.Synthesis of oxazolidinone derivatives and their coagulation factor Ⅹ a inhibitory activity[J].Chin J Med Chem,2019,29(6):426
[3] 康凯,曹久妹.利伐沙班抗凝效果评估方法的研究进展[J].诊断学理论与实践,2019,18(2):218
KANG K,CAO JM.Research progress of anticoagulation evaluation method of rivaroxaban[J].J Diagno Concept Pract,2019,18(2):218
[4] 杨健,朱亚楠,刘晓岩,等.新凝血因子Xa 抑制剂Bg 115-2的抗血栓作用及药代动力学[J].中国药理学与毒理学杂志,2012,26(1):10
YANG J,ZHU YN,LIU XY,et al.Antithrombotic effect and pharmacokinetics of bg115-2,a new clotting factor Xa inhibitor[J].Chin J Pharmacol Toxicol,2012,26(1):10
[5] 郑文灿,法艳梅,吕亚青,等.利伐沙班在临床治疗中的研究进展[J].中国处方药,2020,18(1):18
ZHENG WC,FA YM,LÜ YQ,et al.Research progress of rivaroxaban in clinical treatment[J].J China Prescr Drug,2020,18(1):18
[6] 陈桂浩,杨跃进.利伐沙班在特殊人群房颤中的研究进展[J].中华医药杂志,2017,97(26):2078
CHEN GH;YANG YJ.Research progress of rivaroxaban in atrial fibrillation of special population[J].Nat Med J China,2017,97(26):2078
[7] 李梅娟,王预立,林杉镱,等.新型口服抗凝药利伐沙班在房颤患者中的应用[J].实用药物与临床,2019,22(10):1009
LI MJ,WANG YL,LIN SY,et al.Application of a new oral anticoagulant rivaroxaban in patients with atrial fibrillation[J].Pract Pharm Clin Remed,2019,22(10):1009
[8] 汪生,杭太俊.药物中基因毒性杂质检测策略的研究[J].中国新药杂志,2019,28(23):2840
WANG S,HANG TJ.Study on the detection strategy of genotoxic impurities in drugs[J].Chin J New Drugs,2019,28(23):2840
[9] 谢含仪,林云良,张瑞凌,等.基因毒性杂质分析方法和前处理技术的研究进展[J].药物分析杂志,2018,38(10):1668
XIE HY,LIN YL,ZHANG RL,et al.Research progress in analysis methods and pretreatment techniques of genotoxic impurities[J].Chin J Pharm Anal,2018,38(10):1668
[10] 王萍,徐彩虹,陈仙,等.原料药中基因毒性杂质控制的研究进展[J].中国现代应用药学,2015,32(1):119
WANG P,XU CH,CHEN X,et al.Research progress in the control of genotoxic impurities in APIs[J].Chin J Mod Appl Pharm,2015,32(1):119
[11] 刘晓丹,陆峰,何伍.浅析遗传毒性杂质在医药工业中的来源和控制[J].药学服务与研究,2017,17(3):235
LIU XD,LU F,HE W.Source and control of genotoxic impurities in pharmaceutical industry[J].Pharm Care Res,2017,17(3):235
[12] 马磊,马玉楠,陈震,等.遗传毒性杂质的警示结构[J].中国新药杂志,2014,23(18):2106
MA L,MA YN,CHEN Z,et al.Warning structure of genotoxic impurities[J].Chin J New Drugs,2014,23(18):2106
[13] 张慧敏,林建群,冯康彪,等.药品中遗传毒性杂质的评估和控制[J].中国现代应用药学,2014,31(9):1160
ZHANG HM,LIN JQ,FENG KB,et al.Evaluation and control of genotoxic impurities in drugs[J].Chin J Mod Appl Pharm,2014,31(9):1160
[14] 戴川,赵圣轩,林世博,等.利伐沙班工业化生产合成路线分析[J].中国药业,2015,24(21):4
DAI C,ZHAO SX,LIN SB,et al.Analysis on the synthetic route of industrial production of rivaroxaban[J].China Pharm,2015,24(21):4
[15] 李果. 利伐沙班合成工艺研究[J].天津化工,2020,34(3):8
LI G.Study on the synthetic technology of rivaroxaban[J].Tianjin Chem Ind,2020,34(3):8
[16] 尹秀娥,胡小燕,侯德粉,等.利伐沙班有关物质的HPLC 测定[J].中国医药工业杂志,2019,50(12):1487
YIN XE,HU XY,HOU DF,et al.Determination of related substances in rivaroxaban by HPLC[J].Chin J Pharm,2019,50(12):1487
[17] RUAN XL,ZHOU J,ZHENG XY,et al.Comparison of the hydrolysis and esterification methods for the determination of genotoxic 5-chlorothiophene-2-carbonyl chloride in rivaroxaban using HPLC[J].Chromatographia,2016,79:413
[18] RAMAN NV,PRASAD AV,REDDY KR.Sensitive derivatization methods for the determination of genotoxic impurities in drug substances using hyphenated techniques[J].J Pharm Biomed Anal,2014,89:276
