Citation: QU Min-Min, CHEN Jia, XU Bin, ZHANG Ya-Jiao, XU Hua, XIE Jian-Wei. A Toxic Effect-Directed Screening Method for Genotoxic Impurities in Drugs Based on Mass Spectrometry Quantitative Analysis of Phosphorylated Histone H2AX[J]. Chinese Journal of Analytical Chemistry, ;2021, 49(9): 1531-1539. doi: 10.19756/j.issn.0253-3820.211096 shu

A Toxic Effect-Directed Screening Method for Genotoxic Impurities in Drugs Based on Mass Spectrometry Quantitative Analysis of Phosphorylated Histone H2AX

State Key Laboratory of Toxicology and Medical Countermeasures and Laboratory of Toxicant Analysis, Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China

Corresponding author: XU Hua, huarxu@163.com
Received Date: 31 January 2021
Revised Date: 6 April 2021

Fund Project: Supported by the National Key R&D Program of China (No.2018YFC1602600) and the National Natural Science Foundation of China(No.21974151).

Inspired by the fact that phosphorylated histone H2AX (γ-H2AX) has emerged as a useful biomarker for break of double-strand DNA. In this study, an isotope dilution-based liquid chromatography-tandem mass spectrometry screening method for genotoxic impurities (GTI) was established. The detection limits for H2AX and γ-H2AX target peptides were 1 ng/mL and 2 ng/mL, respectively. The accuracy and precision could meet the methodological requirement for biological samples analysis. This study focused on two kinds of important genotoxic impurities, ethylmethylsulfone (EMS) and N-nitrosodimethylamine (NDMA), in two human cell lines (HepG2 and HeLa) with different metabolic capabilities. The results showed that EMS had genotoxic characteristics in the two cell lines, while NDMA had genotoxicity only in HepG2 cells. The sensitivities of the method were 0.06 μg/g for EMS and 0.03 μg/g for NDMA, which satisfied the international detection limit standard (EMS:0.6 μg/g, NDMA:0.3 μg/g). The method was further applied to the screening of GTI in five commercially available tablets. The tablets were dissolved in water, followed by incubating with the cells to detect γ-H2AX level. The results showed that other unconcerned ingredients in these drugs did not interfere with the detection. This method was convenient, accurate and sensitive, and could be used for rapid detection of GTI in drugs.
- Mass spectrometry,
- Phosphorylated histone H2AX,
- Quantification,
- Genotoxic impurities,
- Screening
1. [1]
  LIU D Q, SUN M J, KORD A S. J. Pharm. Biomed., 2010, 51(5):999-1014.
2. [2]
  REDDY A V B, JAAFAR J, UMAR K, MAJID Z A, ARISA B, TALIB J. J. Sep. Sci., 2015, 38(5):764-779.
3. [3]
  BENIGNI R, BOSSA C. Curr. Comput. Aid. Drug, 2006, 2(2):169-176.
4. [4]
  EMEA. European Medicines Agency announces recall of Viracept.[EB/OL].
5. [5]
  EMEA. EMA reviewing medicines containing valsartan from Zhejiang Huahai following detection of an impurity. EMA/459276/2018.
6. [6]
  ICH M7(R1). Assessment and Control of DNA Reactive (Mutagenic) Impurities in Pharmaceuticals to Limit Potential Carcinogenic Risk. 2017.[EB/OL]. http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Multidisciplinary/M7/M7_R1_Addendum_Step_4_2017_0331.pdf.
7. [7]
  EMEA. Guideline on the Limit of Genotoxic Impurities[EB/OL]. http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2009/09/WC500002903.pdf.
8. [8]
  FDA Guidance for Industry Specific Aspects of Regulatory GenotoxicityTests for Pharmaceuticals.[EB/OL]. https://www.fda.gov/downloads/drugs/guidancecomplianceregulatoryinformation/guidances/ucm074925.pdf.
9. [9]
  National Medical Products Administration. Technical Guidelines for Research on Nitrosamine Impurities in Chemical Drugs (for Trial Implementation). 2020.[2020-05-17]. http://www.nmpa.gov.cn/WS04/CL2138/377043.html.
10. [10]
  KAKADIYA P R, CHANDRASHEKHAR T G, GANGULY S, SINGH D K, SINGH V. J. Pharmaceut. Biomed., 2011, 6(2):21-28.
11. [11]
12. [12]
13. [13]
  ZHOU J, XU J, ZHENG X, LIU W, ZHENG F. J. Sep. Sci., 2017, 40(17):3414-3421.
14. [14]
  ELDER D P, SNODIN D J. J. Pharm. Pharmacol., 2009, 61(3):269-278.
15. [15]
16. [16]
  KOPP B, KHOURY L, AUDEBERT M. Arch. Toxicol., 2019, 98(8):2103-2114.
17. [17]
  LEE Y, WANG Q, SHURYAK I, BRENNER DJ, TURNER H C. Radiat. Oncol., 2019, 14(150):1-10.
18. [18]
19. [19]
  SNODIN D J, MCCROSSEN S D. Regul. Toxicol. Pharmacol., 2013, 67(2):299-316.
20. [20]
  SMART D J, AHMEDI KP, HARVEY J S, LYNCH A M. Mutat. Res., 2011, 715(1-2):25-31.
21. [21]
  GEORGE J, TSUCHISHIMA M, TSUTSUMI M. Cell Death Dis., 2019, 10(1):1801-1809.
22. [22]
  PIOTROWSKA H, KUCINSKA M, MURIAS M. Mol. Cell. Biochem., 2013, 383(1-2):95-102.
23. [23]
24. [24]
25. [25]
1. [1]
  Wei Shao , Wanqun Zhang , Pingping Zhu , Wanqun Hu , Qiang Zhou , Weiwei Li , Kaiping Yang , Xisheng Wang . Design and Practice of Ideological and Political Cases in the Course of Instrument Analysis Experiment: Taking the GC-MS Experiment as an Example. University Chemistry, 2024, 39(2): 147-154. doi: 10.3866/PKU.DXHX202309048
2. [2]
  Wei Peng , Baoying Wen , Huamin Li , Yiru Wang , Jianfeng Li . Exploration and Practice on Raman Scattering Spectroscopy Experimental Teaching. University Chemistry, 2024, 39(8): 230-240. doi: 10.3866/PKU.DXHX202312062
3. [3]
  Zian Lin , Yingxue Jin . Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS) for Disease Marker Screening and Identification: A Comprehensive Experiment Teaching Reform in Instrumental Analysis. University Chemistry, 2024, 39(11): 327-334. doi: 10.12461/PKU.DXHX202403066
4. [4]
  Ling Bai , Limin Lu , Xiaoqiang Wang , Dongping Wu , Yansha Gao . Exploration and Practice of Teaching Reforms in “Quantitative Analytical Chemistry” under the Perspective of New Agricultural Science. University Chemistry, 2024, 39(3): 158-166. doi: 10.3866/PKU.DXHX202308101
5. [5]
  Wei Li , Jinfan Xu , Yongjun Zhang , Ying Guan . 共价有机框架整体材料的制备及食品安全非靶向筛查应用——推荐一个仪器分析综合化学实验. University Chemistry, 2025, 40(6): 276-285. doi: 10.12461/PKU.DXHX202406013
6. [6]
  Mi Wen , Baoshuo Jia , Yongqi Chai , Tong Wang , Jianbo Liu , Hailong Wu . Improvement of Fluorescence Quantitative Analysis Experiment: Simultaneous Determination of Rhodamine 6G and Rhodamine 123 in Food Using Chemometrics-Assisted Three-Dimensional Fluorescence Method. University Chemistry, 2025, 40(4): 390-398. doi: 10.12461/PKU.DXHX202405147
7. [7]
  Zunxiang Zeng , Yuling Hu , Yufei Hu , Hua Xiao . Analysis of Plant Essential Oils by Supercritical CO₂Extraction with Gas Chromatography-Mass Spectrometry: An Instrumental Analysis Comprehensive Experiment Teaching Reform. University Chemistry, 2024, 39(3): 274-282. doi: 10.3866/PKU.DXHX202309069
8. [8]
  Xinran Zhang , Siqi Liu , Yichi Chen , Qingli Zou , Qinghong Xu , Yaqin Huang . From Protein to Energy Storage Materials: Edible Gelatin Jelly Electrolyte. University Chemistry, 2025, 40(7): 255-266. doi: 10.12461/PKU.DXHX202408104
9. [9]
  Min Gu , Huiwen Xiong , Liling Liu , Jilie Kong , Xueen Fang . Rapid Quantitative Detection of Procalcitonin by Microfluidics: An Instrumental Analytical Chemistry Experiment. University Chemistry, 2024, 39(4): 87-93. doi: 10.3866/PKU.DXHX202310120
10. [10]
  Hao Wu , Zhen Liu , Dachang Bai . ¹H NMR Spectrum of Amide Compounds. University Chemistry, 2024, 39(3): 231-238. doi: 10.3866/PKU.DXHX202309020
11. [11]
  Cheng Zheng , Shiying Zheng , Yanping Zhang , Shoutian Zheng , Qiaohua Wei . Synthesis, Copper Content Analysis, and Luminescent Performance Study of Binuclear Copper (I) Complexes with Isomeric Luminescence Shift: A Comprehensive Chemical Experiment Recommendation. University Chemistry, 2024, 39(7): 322-329. doi: 10.3866/PKU.DXHX202310131
12. [12]
  Yuhao SUN , Qingzhe DONG , Lei ZHAO , Xiaodan JIANG , Hailing GUO , Xianglong MENG , Yongmei GUO . Synthesis and antibacterial properties of silver-loaded sod-based zeolite. Chinese Journal of Inorganic Chemistry, 2024, 40(4): 761-770. doi: 10.11862/CJIC.20230169
13. [13]
  Pei Li , Yuenan Zheng , Zhankai Liu , An-Hui Lu . Boron-Containing MFI Zeolite: Microstructure Control and Its Performance of Propane Oxidative Dehydrogenation. Acta Physico-Chimica Sinica, 2025, 41(4): 100034-. doi: 10.3866/PKU.WHXB202406012
14. [14]
  Jiali CHEN , Guoxiang ZHAO , Yayu YAN , Wanting XIA , Qiaohong LI , Jian ZHANG . Machine learning exploring the adsorption of electronic gases on zeolite molecular sieves. Chinese Journal of Inorganic Chemistry, 2025, 41(1): 155-164. doi: 10.11862/CJIC.20240408
15. [15]
  Yiping HUANG , Liqin TANG , Yufan JI , Cheng CHEN , Shuangtao LI , Jingjing HUANG , Xuechao GAO , Xuehong GU . Hollow fiber NaA zeolite membrane for deep dehydration of ethanol solvent by vapor permeation. Chinese Journal of Inorganic Chemistry, 2025, 41(2): 225-234. doi: 10.11862/CJIC.20240224
16. [16]
  Jiandong Liu , Zhijia Zhang , Mikhail Kamenskii , Filipp Volkov , Svetlana Eliseeva , Jianmin Ma . Research Progress on Cathode Electrolyte Interphase in High-Voltage Lithium Batteries. Acta Physico-Chimica Sinica, 2025, 41(2): 100011-. doi: 10.3866/PKU.WHXB202308048
17. [17]
  Xianyong Lu , Tao Hu . Developing an Innovative Inorganic Chemistry Teaching Model Based on Aerospace Specialty Characteristics. University Chemistry, 2025, 40(7): 127-131. doi: 10.12461/PKU.DXHX202409037
18. [18]
  Chongjing Liu , Yujian Xia , Pengjun Zhang , Shiqiang Wei , Dengfeng Cao , Beibei Sheng , Yongheng Chu , Shuangming Chen , Li Song , Xiaosong Liu . Understanding Solid-Gas and Solid-Liquid Interfaces through Near Ambient Pressure X-Ray Photoelectron Spectroscopy. Acta Physico-Chimica Sinica, 2025, 41(2): 100013-. doi: 10.3866/PKU.WHXB202309036
19. [19]
  Zhaoxuan ZHU , Lixin WANG , Xiaoning TANG , Long LI , Yan SHI , Jiaojing SHAO . Application of poly(vinyl alcohol) conductive hydrogel electrolytes in zinc ion batteries. Chinese Journal of Inorganic Chemistry, 2025, 41(5): 893-902. doi: 10.11862/CJIC.20240368
20. [20]
  Changsheng An , Tao Liu . Decoding SEI chemistry at the lithium-metal potential. Acta Physico-Chimica Sinica, 2025, 41(9): 100101-0. doi: 10.1016/j.actphy.2025.100101

Get Citation

PDF

XML

Metrics

PDF Downloads(9)
Abstract views(1003)
HTML views(207)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142