{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T03:33:15Z","timestamp":1792812795347,"version":"4.5.0"},"reference-count":33,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2025,10,4]],"date-time":"2025-10-04T00:00:00Z","timestamp":1759536000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Key Research and Development Project of Shandong Province","award":["2024CXGC010618"],"award-info":[{"award-number":["2024CXGC010618"]}]},{"name":"Jinan Haiyou Industry Leading Talent Support Project","award":["CYLJ20241901438"],"award-info":[{"award-number":["CYLJ20241901438"]}]},{"name":"Taishan Industrial Leading Talents Program","award":["TSCX202306076"],"award-info":[{"award-number":["TSCX202306076"]}]}],"content-domain":{"domain":["www.mdpi.com"],"crossmark-restriction":true},"short-container-title":["Molecules"],"abstract":"<jats:p>DNA direct reactive impurities (DDRIs) can react with nucleophilic sites of DNA, leading to mutations. The control strategies outlined in International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) M7 are based on the known compound structure of DDRIs. Non-target screening of DDRIs in drugs is still challenging due to the diversity of the species and the poor stability. In this study, a derivatization reagent including a reactive group and report group was designed to screen DDRIs. Based on the electrophilic theory of chemical carcinogenesis, an amine reagent was used as a reactive group to interact with DDRIs. Two derivatization reagents, p-methoxyaniline and p-methoxybenzoyl-\u03b2-alaninamide, were employed, each containing different chromatographic modification groups to mitigate matrix effects. The derivatization products were analyzed by ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry (UPLC-HRMS). Non-target screening for DDRIs was achieved by product ions filtering of the report group.<\/jats:p>","DOI":"10.3390\/molecules30193981","type":"journal-article","created":{"date-parts":[[2025,10,6]],"date-time":"2025-10-06T13:33:41Z","timestamp":1759757621000},"page":"3981","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Derivative-Based Non-Target Identification of DNA-Reactive Impurities with Fragment Ion Filtering"],"prefix":"10.3390","volume":"30","author":[{"given":"Dongmei","family":"Zhang","sequence":"first","affiliation":[{"name":"Biological Products Research Division, Shandong Institute for Food and Drug Control, Xinluo Avenue No. 2749, Jinan 250101, China"},{"name":"National Medical Products Administration Key Laboratory for Research and Evaluation of Generic Drugs, Jinan 250101, China"},{"name":"Industrial Technology Foundation Public Service Platform, Jinan 250101, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5105-918X","authenticated-orcid":false,"given":"Baojian","family":"Hang","sequence":"additional","affiliation":[{"name":"Biological Products Research Division, Shandong Institute for Food and Drug Control, Xinluo Avenue No. 2749, Jinan 250101, China"},{"name":"National Medical Products Administration Key Laboratory for Research and Evaluation of Generic Drugs, Jinan 250101, China"},{"name":"Industrial Technology Foundation Public Service Platform, Jinan 250101, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiran","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pengfei","family":"You","sequence":"additional","affiliation":[{"name":"Biological Products Research Division, Shandong Institute for Food and Drug Control, Xinluo Avenue No. 2749, Jinan 250101, China"},{"name":"National Medical Products Administration Key Laboratory for Research and Evaluation of Generic Drugs, Jinan 250101, China"},{"name":"Industrial Technology Foundation Public Service Platform, Jinan 250101, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-3504-2910","authenticated-orcid":false,"given":"Feng","family":"Shi","sequence":"additional","affiliation":[{"name":"Biological Products Research Division, Shandong Institute for Food and Drug Control, Xinluo Avenue No. 2749, Jinan 250101, China"},{"name":"National Medical Products Administration Key Laboratory for Research and Evaluation of Generic Drugs, Jinan 250101, China"},{"name":"Industrial Technology Foundation Public Service Platform, Jinan 250101, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-3777-4307","authenticated-orcid":false,"given":"Liping","family":"Gong","sequence":"additional","affiliation":[{"name":"Biological Products Research Division, Shandong Institute for Food and Drug Control, Xinluo Avenue No. 2749, Jinan 250101, China"},{"name":"National Medical Products Administration Key Laboratory for Research and Evaluation of Generic Drugs, Jinan 250101, China"},{"name":"Industrial Technology Foundation Public Service Platform, Jinan 250101, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2507","DOI":"10.1021\/cr100222q","article-title":"Mechanisms of chemical carcinogenicity and mutagenicity: A review with implications for predictive toxicology","volume":"111","author":"Benigni","year":"2011","journal-title":"Chem. Rev."},{"key":"ref_2","unstructured":"ICH Guideline (2013). Assessment and Control of DNA Reactive (Mutagenic) Impurities in Pharmaceuticals to Limit Potential Carcinogenic Risk, ICH Guideline. Available online: https:\/\/www.ich.org\/page\/multidisciplinary-guidelines."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1186\/1742-6405-6-18","article-title":"Elevated ethyl methanesulfonate (EMS) in nelfinavir mesylate (Viracept, Roche): Overview","volume":"6","author":"Pozniak","year":"2009","journal-title":"AIDS Res. Ther."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1869","DOI":"10.1016\/j.talanta.2008.09.041","article-title":"Development and validation of LC methods with visible detection using pre-column derivatization and mass detection for the assay of voglibose","volume":"77","author":"Ramakrishna","year":"2009","journal-title":"Talanta"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"25797","DOI":"10.1039\/C9RA03835C","article-title":"A new HPLC-UV derivatization approach for the determination of potential genotoxic benzyl halides in drug substances","volume":"9","author":"Ji","year":"2019","journal-title":"RSC Adv."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"873","DOI":"10.1002\/rcm.7505","article-title":"Applicability of gas chromatography\/quadrupole-Orbitrap mass spectrometry in support of pharmaceutical research and development","volume":"30","author":"Baldwin","year":"2016","journal-title":"Rapid Commun Mass Spectrom."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Lee, K., Yoo, W., and Jeong, J.H. (2022). Analytical Method Development for 19 Alkyl Halides as Potential Genotoxic Impurities by Analytical Quality by Design. Molecules, 27.","DOI":"10.3390\/molecules27144437"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1177\/02611929241256040","article-title":"In Silico Phototoxicity Prediction of Drugs and Chemicals by using Derek Nexus and QSAR Toolbox","volume":"52","author":"Ahuja","year":"2024","journal-title":"Altern. Lab. Anim."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Zhou, X., Chen, X., Fan, L., Dong, H., Ren, Y., and Chen, X. (2022). Stepwise Diagnostic Product Ions Filtering Strategy for Rapid Discovery of Diterpenoids in Scutellaria barbata Based on UHPLC-Q-Exactive-Orbitrap-MS. Molecules, 27.","DOI":"10.3390\/molecules27238185"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"6049","DOI":"10.1039\/D1AN01047F","article-title":"Overview, consequences, and strategies for overcoming matrix effects in LC-MS analysis: A critical review","volume":"146","author":"Nasiri","year":"2021","journal-title":"Analyst"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Cortese, M., Gigliobianco, M.R., Magnoni, F., Censi, R., and Di Martino, P.D. (2020). Compensate for or minimize matrix effects? Strategies for overcoming matrix effects in liquid chromatography-mass spectrometry technique: A tutorial review. Molecules, 25.","DOI":"10.3390\/molecules25133047"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1375","DOI":"10.1007\/s00216-011-4901-y","article-title":"A new approach for generic screening and quantitation of potential genotoxic alkylation compounds by pre-column derivatization and LC-MS\/MS analysis","volume":"400","author":"Beerman","year":"2011","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.talanta.2012.11.012","article-title":"Diagnostic fragment-ion-based and extension strategy coupled to DFIs intensity analysis for identification of chlorogenic acids isomers in Flos Lonicerae Japonicae by HPLC-ESI-MS(n)","volume":"104","author":"Zhang","year":"2013","journal-title":"Talanta"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"14528","DOI":"10.1021\/acs.analchem.0c02719","article-title":"In-depth annotation strategy of saturated hydroxy fatty acids based on their chromatographic retention behaviors and MS fragmentation patterns","volume":"92","author":"Zhu","year":"2020","journal-title":"Anal. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"385","DOI":"10.2174\/1389201024666230726152629","article-title":"Genotoxic Impurities in Critical Analysis of Product Development: Recent Advancements, Patents, and Current Challenges","volume":"25","author":"Swain","year":"2024","journal-title":"Curr. Pharm. Biotechnol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1056","DOI":"10.1016\/j.yrtph.2024.105669","article-title":"Retrospective application of ICH M7 to anti-hypertensive drugs in Brazil: Risk assessment of potentially mutagenic impurities","volume":"151","author":"Waechter","year":"2024","journal-title":"Regul. Toxicol. Pharmacol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"524","DOI":"10.1038\/s41370-023-00574-6","article-title":"Non-targeted analysis (NTA) and suspect screening analysis (SSA): A review of examining the chemical exposome","volume":"33","author":"Manz","year":"2023","journal-title":"J. Expo. Sci. Environ. Epidemiol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Basu, A.K. (2018). DNA Damage, Mutagenesis and Cancer. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19040970"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1021\/acs.chemrestox.3c00302","article-title":"Mass Spectral Library for DNA Adductomics","volume":"37","author":"Walmsley","year":"2024","journal-title":"Chem. Res. Toxicol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"bgaf008","DOI":"10.1093\/carcin\/bgaf008","article-title":"Reactions of [13C]-labelled tobacco smoke with DNA to generate selected adducts formed without metabolic activation","volume":"46","author":"Tang","year":"2025","journal-title":"Carcinogenesis"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1584\/jpestics.D23-060","article-title":"Investigation of variability in the matrix effect on stable isotope-labeled internal standards in liquid chromatography-tandem mass spectrometry analysis of 25 pesticides in vegetables","volume":"49","author":"Banno","year":"2024","journal-title":"J. Pestic. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"465580","DOI":"10.1016\/j.chroma.2024.465580","article-title":"Matrix effect evaluation using multi-component post-column infusion in untargeted hydrophilic interaction liquid chromatography-mass spectrometry plasma metabolomics","volume":"1740","author":"Zhu","year":"2025","journal-title":"J. Chromatogr. A"},{"key":"ref_23","first-page":"3997","article-title":"Orthogonal Deprotection Strategy of Fmoc Provides Improved Synthesis of Sensitive Peptides: Application to Z-Arg-Lys-AOMK","volume":"9","author":"Almaliti","year":"2024","journal-title":"ACS Omega"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2149","DOI":"10.1021\/ja00897a025","article-title":"Solid Phase Peptide Synthesis. I. The Synthesis of a Tetrapeptide","volume":"85","author":"Merrifield","year":"1963","journal-title":"J. Am. Chem. Soc."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Reichardt, C., and Welton, T. (2011). Solvents and Solvent Effects in Organic Chemistry, Wiley-VCH Verlag GmbH & Co. KGaA.","DOI":"10.1002\/9783527632220"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Day, D.M., Farmer, T.J., Granelli, J., Lofthouse, J.H., Lynch, J., McElroy, C.R., Sherwood, J., Shimizu, S., and Clark, J.H. (2022). Reaction Optimization for Greener Chemistry with a Comprehensive Spreadsheet Tool. Molecule, 27.","DOI":"10.3390\/molecules27238427"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1199","DOI":"10.1002\/jcc.27079","article-title":"Electronic and magnetic properties of TATA-DNA sequence driven by chemical functionalization","volume":"44","year":"2023","journal-title":"J. Comput. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"8702","DOI":"10.1093\/nar\/gkae639","article-title":"Site-specific N-alkylation of DNA oligonucleotide nucleobases by DNAzyme-catalyzed reductive amination","volume":"52","author":"Boyd","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1910","DOI":"10.1039\/D3OB00070B","article-title":"Defining the substrate scope of DNAzyme catalysis for reductive amination with aliphatic amines","volume":"21","author":"Yang","year":"2023","journal-title":"Org. Biomol. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1021\/ci030424n","article-title":"Chemical reactivity as a tool to study carcinogenicity: Reaction between estradiol and estrone 3,4-quinones ultimate carcinogens and guanine","volume":"44","author":"Huetz","year":"2004","journal-title":"J. Chem. Inf. Comput. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4078","DOI":"10.1021\/jo060098l","article-title":"Chemical reactivity as a tool to study carcinogenicity: Reaction between chloroethylene oxide and guanine","volume":"71","author":"Bren","year":"2006","journal-title":"J. Org. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1016\/j.tiv.2012.07.009","article-title":"Can the chemical reactivity of an ultimate carcinogen be related to its carcinogenicity? An application to propylene oxide","volume":"27","author":"Mavri","year":"2013","journal-title":"Toxicol. Vitr."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"4383","DOI":"10.1021\/acs.analchem.0c03931","article-title":"Simple and Sensitive Method for Deep Profiling of Host Cell Proteins in Therapeutic Antibodies by Combining Ultra-Low Trypsin Concentration Digestion, Long Chromatographic Gradients, and BoxCar Mass Spectrometry Acquisition","volume":"93","author":"Nie","year":"2021","journal-title":"Anal. Chem."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/30\/19\/3981\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,6]],"date-time":"2025-10-06T13:34:59Z","timestamp":1759757699000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/30\/19\/3981"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,4]]},"references-count":33,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2025,10]]}},"alternative-id":["molecules30193981"],"URL":"https:\/\/doi.org\/10.3390\/molecules30193981","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,4]]}}}