Citation: QU Min-Min, CHEN Jia, XU Bin, LI Zhi, GUO Lei, XU Hua, XIE Jian-Wei. Determination of Genotoxic Compounds with Different Modes of Action on Histone H3 Acetylation by Protein Modification Quantitative Mass Spectrometry[J]. Chinese Journal of Analytical Chemistry, ;2022, 50(6): 940-947. doi: 10.19756/j.issn.0253-3820.221025 shu

Determination of Genotoxic Compounds with Different Modes of Action on Histone H3 Acetylation by Protein Modification Quantitative Mass Spectrometry

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

Corresponding author: XU Hua, huarxu@163.com
Received Date: 14 January 2022
Revised Date: 31 March 2022

Fund Project: Supported by the National Natural Science Foundation of China(No. 21974151).

Three target peptides, H3K9ac, H3K14ac and H3, in human cell lines were determined by highperformance liquid chromatography coupled with triple quadrupole mass spectrometry under positive ion multiple reaction monitoring mode(MRM). The cell samples were pre-treated by trypsin digestion,propionylation derivatization, desalination, peptide enrichment, and then separated by ACQUITY UPLC BEH C₁₈ column. The results indicated that the three peptides in cells had good linear relationship within the concentration range of 1-250 ng/mL(R²>0.99). The detection limits(S/N=3) of the peptides were 0.1 ng/mL.Accuracy, precision, matrix effects and recoveries meet the methodological requirement for biological samples analysis. The established method had many advantages such as simple operation, high sensitivity,fast detection and accurate quantification, which had been initially applied to evaluate the effects of two classic genotoxic compounds(camptothecin and colchicine) with different modes of action on the acetylation at position 9 and position 14 of histone H3 in cells. This method provided important technical support for DNA damage/repair and transcription research of genotoxic compounds.
- High performance liquid chromatography,
- Mass spectrometry,
- Histone H3,
- Acetylation,
- Genotoxic compounds
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