Objective: To establish an HPLC-MS/MS method for the determination of the genotoxic impurities (S)-5-chloro-N-(3-chloro-2-hydroxypropyl) thiophene-2-amide and 4-(3-oxy-4-morpholine) nitrobenzene in rivaroxaban. The factors affecting impurities separation and the response of mass spectrometry were systematically investigated. Methods: A ZORBAX SB-C18 column(150 mm×2.1 mm, 3.5 μm) was used with 10 mmol·L-1 ammonium formate solution-methanol as mobile phase at flow rate of 0.4 mL·min-1. And MRM detection by electrospray positive ionization was used. Results: (S)-5-chloro-N-(3-chloro-2-hydroxypropyl) thiophene-2-amide and 4-(3-oxy-4-morpholine) nitrobenzene had good linear relationships in the range of 0.008-3.4 ng·mL-1 and 0.2-3.4 ng·mL-1, respectively. The average recoveries(n=9) were 97.9% and 98.9%, RSD were 0.94% and 0.68% respectively. The limits of quantitation were 0.008 ng·mL-1 and 0.2 ng·mL-1 respectively. The results of genotoxic impurity residue determination in 15 batches of rivaroxaban samples were far below the limit value. Conclusion: The method is sensitive and specific, and can effectively detect trace genotoxic impurities in rivaroxaban.
LI Ping, ZHANG Feng-mei, ZHOU Ping, CAO Yue-feng, WANG Jian
. Detection of trace genotoxic impurities in rivaroxaban by HPLC-MS/MS*[J]. Chinese Journal of Pharmaceutical Analysis, 2024
, 44(1)
: 101
-108
.
DOI: 10.16155/j.0254-1793.2024.01.10
[1] 王开. 利伐沙班片的处方和工艺开发[D]. 杭州: 浙江大学, 2020
WANG K. Formulation and Process Development of Rivaroxaban Tablets[D]. Hangzhou: Zhejiang University, 2020
[2] 白龙, 李洁. 活血通肺汤联合利伐沙班片治疗急性肺动脉栓塞的疗效观察[J]. 血栓与止血学, 2022, 28(3): 385
BAI L, LI J. Clinical observation of Huoxue Tongfei decoction combined with rivaroxaban tablets in the treatment of acute pulmonary embolism[J]. Chin J Thromb Hemost, 2022, 28(3): 385
[3] 朱道平. 在静脉血栓的治疗中采取利伐沙班片治疗的临床效果分析[J]. 系统医学, 2021, 6(20): 156
ZHU DP. Analysis of the clinical effect of rivaroxaban tablets in the treatment of venous thrombosis[J]. Syst Med, 2021, 6(20): 156
[4] 王欢, 曾莉娜, 姚松, 等. 利伐沙班片在血栓栓塞性疾病中的应用进展[J]. 临床误诊误治, 2020, 33(10): 109
WANG H, ZENG LN, YAO S, et al. Application progress of rivaroxaban tablets in thromboembolic diseases[J]. Clin Misdiagn Misther, 2020, 33(10): 109
[5] 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. Strategies for determination of genotoxic impurities in pharmaceuticals[J]. Chin J New Drugs, 2019, 28(23): 2840
[6] 徐洁. 利伐沙班中苯胺类潜在基因毒性杂质的HPLC法测定[J]. 药物分析杂志, 2017, 37(1): 105
XU J. Determination of aniline potential genotoxic impurity in Rivaroxaban by HPLC[J]. Chin J Pharm Anal, 2017, 37(1): 105
[7] 陈轶嘉, 石玲玲, 陈阳, 等. 无催化剂衍生GC-MS法测定利伐沙班基因毒杂质5-氯-2-酰氯噻吩[J]. 药物分析杂志, 2021, 41(4): 714
CHEN YJ, SHI LL, CHEN Y, et al. Determination of genotoxic 5-chlorothiophene-2-carbonyl chloride in rivaroxaban by GC-MS with catalyst-free derivatization[J]. Chin J Pharm Anal, 2021, 41(4): 714
[8] 冯贵成. 抗凝血药物利伐沙班合成工艺研究[D]. 杭州: 浙江工业大学, 2017
FENG GC. Study of the Synthesis Process of Antithrombotic Drug Rivaroxaban[D]. Hangzhou: Zhejiang University of Technology, 2017
[9] 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
[10] 王倩倩, 顾铭盼, 沈志超. HPLC法测定利伐沙班中基因杂质5-氯噻吩-2-甲酰氯[J]. 广东化工, 2019, 46(3): 176
WANG QQ, GU MP, SHEN ZC. Methods for the determination of genotoxic 5-chlorothiophene-2-carbonyl chloride in rivaroxaban using HPLC[J]. Guangdong Chem Ind, 2019, 46(3): 176
[11] BARSAM SJ, PATEL JP, ROBERTS LN, et al. The impact of body weight on rivaroxaban pharmacokinetics[J]. Res Pract Thromb Haemost, 2017, 1(2): 180
[12] 张云峰, 钱建钦, 王建. HPLC-MS/MS法分析氟胞嘧啶中痕量基因毒性杂质N,N-二甲基苯胺[J]. 药物分析杂志, 2017, 37(2): 265
ZHANG YF, QIAN JQ, WANG J. Determination of trace level of genotoxic impurity N,N-dimethylaniline in flucytosine by HPLC-MS/MS[J]. Chin J Pharm Anal, 2017, 37(2): 265
[13] 王建, 周萍, 刘贵君, 等. HPLC-MS/MS分析阿哌沙班中基因毒性杂质及基质效应消除研究[J]. 中国现代应用药学, 2022, 39(10): 1336
WANG J, ZHOU P, LIU GJ, et al. Study on genotoxic impurities and elimination of matrix effect in apixaban by HPLC-MS/MS[J]. Chin J Mod Appl Pharm, 2022, 39(10): 1336
[14] EMEA. Guideline on the Limits of Genotoxic Impurities[R]. 2006: 2
[15] ICH Q2(R1). Current Step4 Version: Parent Guideline Dated 27 October 1994[S]. 1994
