Citation: WU Hai-Jiang, ZHANG Ya-Jiao, LIU Qin, CHEN Jia, XU Bin, XU Hua, XIE Jian-Wei. Comparison of Different Hydrolysis Methods in DNA Adducts Analysis and Application[J]. Chinese Journal of Analytical Chemistry, ;2022, 50(7): 1048-1056. doi: 10.19756/j.issn.0253-3820.221018 shu

Comparison of Different Hydrolysis Methods in DNA Adducts Analysis and Application

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

Corresponding author: XIE Jian-Wei, xiejianwei@bmi.ac.cn
Received Date: 12 January 2022
Revised Date: 30 March 2022

Fund Project: Supported by the National Natural Science Foundation of China (No.22104153).

The identification and quantification of DNA adducts is an important technical issue in the research of DNA damage, epigenetics and genotoxic impurities screening. Various DNA hydrolysis methods with individual advantages are available for different applicable analytical purposes to produce bases, nucleosides or nucleic acid fragments. By taking the classic alkylation reagent mustard gas as a model compound, three common DNA hydrolysis methods including thermal hydrolysis, acid hydrolysis and enzymatic hydrolysis were comparatively used to pretreat the sample from in vitro incubation solution or in vivo animal exposure experiments. The produced DNA adducts were screened and quantitatively detected by mass spectrometry. Hydrolysis efficiency of DNA adducts in different analytical samples as well as different sites and adduct forms were investigated. Results showed that there were differences in the stability of DNA adducts derived from different base modification sites. The N-site adducts of guanine and adenine were relatively stable under each hydrolysis condition, while O6 site adducts of guanine were unstable to acid and thermal hydrolysis. In comparison, the efficiency of thermal hydrolysis was lower, while the acid hydrolysis was more efficient which improved 10.4%-94.9% hydrolysis yields and DNA-base adducts were more than 99% among the products, thus suitable for the quantitative analysis of structure-known DNA adducts by mass spectrometry. For enzymatic hydrolysis method, it could produce various adduct forms, such as base- and/or nucleoside-adducts, i.e., the product information was rich, which was suitable for screening and identification of structure-unknown adducts. However, the enzymatic hydrolysis efficiency was often affected by the factors such as DNA modification site, matrix effect and enzymatic hydrolysis conditions which might consequently affect the accuracy of DNA adduct quantification.
- DNA hydrolysis,
- DNA adducts,
- Liquid chromatography-tandem mass spectrometry
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