Archives of Toxicology最新文献

{"title":"A simplified LC–MS-based method for sensitive analysis of DNA adducts utilizing accessible in vitro metabolism models","authors":"Andrea Gerdemann,&nbsp;Matthias Behrens,&nbsp;Georgia Günther,&nbsp;Ahmed Ghallab,&nbsp;Jan G. Hengstler,&nbsp;Hans-Ulrich Humpf,&nbsp;Melanie Esselen","doi":"10.1007/s00204-025-04125-w","DOIUrl":"10.1007/s00204-025-04125-w","url":null,"abstract":"<div><p>Genetic information of living cells is encoded by the specific arrangement of nucleobases in deoxyribonucleic acid (DNA). Even minor changes or modifications of these nucleobases, favored by many reactive sites, can significantly affect correct replication and DNA integrity leading to severe toxic effects. Analyzing these DNA modifications is highly complex and often requires non-specific assays for DNA damage or ³²<i>P</i>-postlabelling. A major limitation of these methods is their inability to provide structural information, a gap that can be addressed by instrumental analytical techniques. An additional major challenge is the selection of an appropriate biological system capable of reliable DNA adduct formation in high yields, since most compounds require metabolic activation prior to reacting with DNA. Therefore, the aim of this study was to develop a fast and simple workflow for sensitive DNA adduct analysis. In addition to highlighting the main pitfalls in sample preparation, this publication focuses on the comparison of biological systems in terms of metabolic activity, using six well-known carcinogens from different chemical classes. The combination of HepG2 cells and liver S9 fractions demonstrated comparable or even superior capabilities to primary hepatocytes and enabled the detection of DNA adducts from aflatoxin B₁, benzo[a]pyrene, methyleugenol as well as α-asarone and β-asarone, particularly after metabolic activation by the aryl hydrocarbon receptor agonist β-naphthoflavone. Notably, the adduct formation of phenylpropanoids was documented for the first time in a non-transfected cancer cell line using high-performance liquid chromatography coupled to mass spectrometry. Therefore, the method can be used to detect previously unknown DNA adducts from diverse chemical classes and provides structural insights into the formed DNA adducts.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":"99 10","pages":"4021 - 4034"},"PeriodicalIF":6.9,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00204-025-04125-w.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144590343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification and characterization of toxic components of Carybdea brevipedalia venom: implications for understanding box jellyfish (Cnidaria: Cubozoa) envenomation","authors":"Du Hyeon Hwang,&nbsp;Ramachandran Loganathan Mohan Prakash,&nbsp;Ravi Deva Asirvatham,&nbsp;Changkeun Kang,&nbsp;Euikyung Kim","doi":"10.1007/s00204-025-04117-w","DOIUrl":"10.1007/s00204-025-04117-w","url":null,"abstract":"<div><p>Jellyfish stings, particularly from cubozoans, pose a significant threat to human health worldwide. However, the major toxins responsible for the clinical symptoms following envenomation by box jellyfish (class Cubozoa) are not well understood. In this study, we investigated the major toxic components of <i>Carybdea brevipedalia</i> venom (CbV) as a representative model of box jellyfish venoms. We identified the key lethal toxin components (CbVLT) from crude CbV. For this, we isolated the toxic components using anion exchange chromatography and fast protein liquid chromatography (FPLC) methods, accompanied by toxicity evaluations. In vitro toxicity assays confirmed the strong cytotoxicity and hemolytic activity of CbVLT. Furthermore, CbVLT exhibited strong lethality in zebrafish, inducing notable hemorrhage and swelling, significant lactate dehydrogenase (LDH) activity, and fatal histopathological changes in vital organs (brain, gills, and heart). These results indicated that CbVLT contains the key lethal toxins in the venom. Subsequently, LC–MS/MS analysis identified 8 toxin homologs in CbVLT, including potassium channel toxins, cytolysins, hemolysins, proteases, and hydrolases. These toxins are responsible for the severe local and systemic reactions and even death caused by <i>C. brevipedalia</i> stings. These findings uncover and characterize the key lethal toxins in CbV, which will help reveal the molecular mechanisms of the venom’s lethality and significantly aid in developing therapeutic strategies to treat stings from the cubozoa jellyfish in the future.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":"99 10","pages":"4263 - 4279"},"PeriodicalIF":6.9,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144574742","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An integrative multi-omics study to identify candidate DNA methylation biomarkers associated with gastric cancer prognosis","authors":"Jingjing Gu,&nbsp;Yanling Wu,&nbsp;Wenyue Tao,&nbsp;Junyi Xin,&nbsp;Hanting Liu,&nbsp;Weida Gong,&nbsp;Qinghong Zhao,&nbsp;Haiyan Chu,&nbsp;Mulong Du,&nbsp;Meilin Wang,&nbsp;Dongmei Wu,&nbsp;Guoquan Tao,&nbsp;Zhengdong Zhang","doi":"10.1007/s00204-025-04118-9","DOIUrl":"10.1007/s00204-025-04118-9","url":null,"abstract":"<div><p>Aberrant DNA methylation (DNAm) is the most well-defined epigenetic hallmark in gastric cancer (GC), which may be associated with a variety of risk factors exposure. In this study, leveraging the multi-omics data of Genome-wide association studies (GWAS), methylation quantitative trait locus (mQTL) and expression quantitative trait locus (eQTL) collected from The Cancer Genome Atlas (TCGA), Genotype-Tissue Expression (GTEx) Project and Gene Expression Omnibus (GEO) database, a joint analysis of cox proportional hazard regression and Summary-data-based Mendelian randomization (SMR) analysis were adapted to investigate the causal associations of DNAm, gene expression and the GC prognosis. The results showed the causal association of hypermethylation of cg16007185, down-regulation of <i>TMX1</i>, and poorer prognosis of GC patients. Mendelian randomization (MR) analysis revealed that exposure to PCB-99, a type of polychlorinated biphenyl, might lead to the hypermethylation of cg16007185. Mediation analysis showed the borderline mediation role of <i>TMX1</i> in the association between cg16007185 and GC survival, with an indirect effect (IE) of 5.24% (<i>P</i> = 0.102). Weighted correlation network analysis (WGCNA) and enrichment analysis predicted that <i>TMX1</i> was involved in the cell proliferation pathway. In vitro experiments validated that promoter hypomethylation could promote the <i>TMX1</i> expression, which inhibited the proliferation and metastatic ability of GC cells. Overall, our results suggest that the hypermethylation of cg16007185, a result of PCB-99 exposure, may promote the poor prognosis of GC patients by decreasing the <i>TMX1</i> expression.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":"99 10","pages":"4067 - 4080"},"PeriodicalIF":6.9,"publicationDate":"2025-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144566947","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Assessing risk of bias in toxicological studies in the era of artificial intelligence","authors":"Thomas Hartung,&nbsp;Sebastian Hoffmann,&nbsp;Paul Whaley","doi":"10.1007/s00204-025-03978-5","DOIUrl":"10.1007/s00204-025-03978-5","url":null,"abstract":"<div><p>Risk of bias is a critical factor influencing the reliability and validity of toxicological studies, impacting evidence synthesis and decision-making in regulatory and public health contexts. The traditional approaches for assessing risk of bias are often subjective and time-consuming. Recent advancements in artificial intelligence (AI) offer promising solutions for automating and enhancing bias detection and evaluation. This article reviews key types of biases—such as selection, performance, detection, attrition, and reporting biases—in in vivo, in vitro, and in silico studies. It further discusses specialized tools, including the SYRCLE and OHAT frameworks, designed to address such biases. The integration of AI-based tools into risk of bias assessments can significantly improve the efficiency, consistency, and accuracy of evaluations. However, AI models are themselves susceptible to algorithmic and data biases, necessitating robust validation and transparency in their development. The article highlights the need for standardized, AI-enabled risk of bias assessment methodologies, training, and policy implementation to mitigate biases in AI-driven analyses. The strategies for leveraging AI to screen studies, detect anomalies, and support systematic reviews are explored. By adopting these advanced methodologies, toxicologists and regulators can enhance the quality and reliability of toxicological evidence, promoting evidence-based practices and ensuring more informed decision-making. The way forward includes fostering interdisciplinary collaboration, developing bias-resilient AI models, and creating a research culture that actively addresses bias through transparent and rigorous practices.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":"99 8","pages":"3065 - 3090"},"PeriodicalIF":6.9,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00204-025-03978-5.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144564311","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gestational vortioxetine exposure impairs pregnancy outcomes and fetal brain development via modulation of BDNF/Bax-Bcl2/5-HT pathways in rats","authors":"Pallavi Singh,&nbsp;Priyanka Agrawal,&nbsp;K. P. Singh","doi":"10.1007/s00204-025-04122-z","DOIUrl":"10.1007/s00204-025-04122-z","url":null,"abstract":"<div><p>Pregnancy presents a critical period during which the use of antidepressants requires cautious evaluation, especially due to the high incidence of depression in women of reproductive age. Vortioxetine hydrobromide (VOX), a newer multimodal antidepressant, is increasingly favored for its improved efficacy and tolerability. However, its safety profile during gestation and its potential long-term impact on fetal brain development remain insufficiently explored. This study aimed to assess the reproductive and developmental neurotoxicity of VOX administered at clinically relevant doses during pregnancy using a Wistar rat model. Pregnant dams received daily oral doses of either 1 mg/kg or 2 mg/kg of VOX from gestation day (GD) 6 to 21. On GD 21, the dams were euthanized, and fetuses along with placental tissues were collected for further analysis. Fetal brains underwent morphological, neurochemical, and histopathological evaluations. Exposure to VOX at the higher dose led to significant maternal and fetal toxicity, marked by reduced maternal weight gain, smaller litter sizes, and decreased fetal and placental weights. Notably, adverse neurodevelopmental outcomes were observed, including reduced fetal brain dimensions, cortical and hippocampal thinning, altered levels of brain-derived neurotrophic factor (BDNF), disrupted monoaminergic neurotransmitter levels (serotonin, dopamine, and norepinephrine), elevated apoptotic markers (Bax/Bcl-2 ratio), and increased acetylcholinesterase activity. These findings suggest that prenatal VOX exposure disrupts essential neurodevelopmental pathways, particularly those involving BDNF and monoaminergic signaling, potentially leading to persistent alterations in brain development. This is the first study to systematically demonstrate the developmental neurotoxicity of VOX, emphasizing the urgent need for further investigation into its safety during pregnancy.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":"99 10","pages":"4243 - 4261"},"PeriodicalIF":6.9,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144564312","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gadolinium toxicity: mechanisms, clinical manifestations, and nanoparticle role","authors":"Jose L. Domingo,&nbsp;Richard C. Semelka","doi":"10.1007/s00204-025-04124-x","DOIUrl":"10.1007/s00204-025-04124-x","url":null,"abstract":"<div><p>Gadolinium-based contrast agents (GBCAs), essential for MRI, are facing renewed scrutiny due to gadolinium (Gd) retention and emerging toxicity profiles. While the link between less stable agents and Nephrogenic Systemic Fibrosis (NSF) in renal impairment is established, gadolinium (Gd) deposition is also observed in the brain, bone, and skin across all GBCA classes, even in patients with normal renal function. This finding has raised concerns and led to a concept of Gadolinium Deposition Disease (GDD). The present review synthesizes current evidence on clinical manifestations and underlying mechanisms. It highlights pathways beyond traditional transmetallation, particularly endogenous nanoparticle formation as a key mechanism for Gd release and retention, potentially challenging the stability assumptions for even macrocyclic agents. Structural factors (linear/macrocyclic; ionic/non-ionic) and stability parameters (thermodynamic log K; kinetic kobs) influencing risk are evaluated alongside regulatory responses. GBCAs should be viewed not as inert diagnostics but as agents with complex, cumulative biological interactions. Future research should focus on developing non-gadolinium alternatives, validating biomarkers for early detection of Gd retention, and conducting controlled trials on chelation therapy efficacy. Clinicians must balance the diagnostic benefits of GBCAs with potential long-term risks, ensuring informed patient consent and judicious use. Innovative approaches, such as Gd-grafted nanodiamonds with high relaxivity and enhanced safety via polyvinylpyrrolidone (PVP) coating, may offer alternatives to traditional GBCAs by reducing toxicity risks<i>.</i> Manganese-based contrast agents, such as Mn-PyC3A, show promise as safer alternatives due to efficient renal and hepatobiliary elimination, even in renal impairment, as demonstrated in rat models.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":"99 10","pages":"3897 - 3916"},"PeriodicalIF":6.9,"publicationDate":"2025-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00204-025-04124-x.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144551776","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Predicting the developmental toxicity of 8-methyl-benzo[a]pyrene (BaP) by physiologically based kinetic (PBK) modeling-facilitated reverse dosimetry and read-across from BaP","authors":"Danlei Wang,&nbsp;Jing Fang,&nbsp;Miaoying Shi,&nbsp;Lenny Kamelia,&nbsp;Ivonne M. C. M. Rietjens,&nbsp;Peter J. Boogaard","doi":"10.1007/s00204-025-04115-y","DOIUrl":"10.1007/s00204-025-04115-y","url":null,"abstract":"<div><p>The present study predicts an in vivo dose-response curve for developmental toxicity of 8-methyl-benzo[a]pyrene (8-MBaP) in rats using PBK model-facilitated reverse dosimetry and read-across from a previously developed model for BaP. As for BaP, in vitro developmental toxicity of 8-MBaP in the mouse embryonic stem cell test (mEST) was shown to require bioactivation to its 3-hydroxy metabolite (3-OH-8-MBaP). To facilitate extrapolation of the in vitro observed developmental toxicity of 3-OH-8-MBaP to an in vivo dose–response curve for 8-MBaP, a PBK model for 8-MBaP was built based on the previously developed PBK model for BaP. The PBK models for BaP and 8-MBaP were validated based on (i) comparison of predicted blood concentrations of BaP or 8-MaP to blood concentrations in rats dosed with BaP or 8-MBaP in the present study and (ii) literature-reported blood concentrations of BaP and its metabolite 3-OH-BaP in rats upon dosing with BaP. The predicted half maximal effect dose (ED₅₀) for developmental toxicity of 8-MBaP that was derived from the predicted in vivo dose–response curve was 1.3-fold lower than that of BaP for which the predicted value was in line with the reported in vivo ED₅₀. The present study illustrates how a new approach methodology (NAM) can be applied to predict developmental toxicity of a methyl substituted PAH, by PBK modeling facilitated read-across from a non-substituted PAH with available in vivo data. The method also elucidates how a methyl substituent could affect the kinetics and subsequent developmental toxicity of a non-substituted PAH.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":"99 10","pages":"4035 - 4050"},"PeriodicalIF":6.9,"publicationDate":"2025-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00204-025-04115-y.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144558906","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A recent decade update on combating doxorubicin-induced toxicities","authors":"Sruthi Sritharan,&nbsp;Nageswaran Sivalingam","doi":"10.1007/s00204-025-04112-1","DOIUrl":"10.1007/s00204-025-04112-1","url":null,"abstract":"<div><p>The gradual rise in global cancer rates every year instills the need for a reliable therapeutic strategy. Chemotherapy alone or in combination with surgery, radiation, or adjuvant therapy is still the preferred treatment option for a wide range of cancers. Doxorubicin (dox) is an extensively used chemotherapeutic anthracycline for combating a broad range of solid and soft-tissue tumors. However, dox therapy is often associated with undesirable side effects, mainly cardiotoxicity, limiting its clinical application. Over the years, many strategies have been developed to overcome these toxicities and improve the drug’s therapeutic potential. This review is focused on discussing the recent investigations into natural compounds, nanoformulations, miRNAs, and physical exercise in protecting against dox-mediated cardiotoxicity, hepatotoxicity, nephrotoxicity, and neurotoxicity.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":"99 9","pages":"3565 - 3578"},"PeriodicalIF":6.9,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144537912","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Long non-coding RNA LINC02499 regulates the expression of CYP3A5 in human liver cells","authors":"Chengcheng Li,&nbsp;Langzhang Li,&nbsp;Siqi Zhang,&nbsp;Yawen Zhang,&nbsp;Tianxing Yang,&nbsp;Liang Li","doi":"10.1007/s00204-025-04120-1","DOIUrl":"10.1007/s00204-025-04120-1","url":null,"abstract":"<div><p>Cytochrome P450 3A5 (CYP3A5) is a critical drug-metabolizing enzyme in human hepatocytes, playing a key role in the metabolism of various clinically relevant drugs, with considerable variability in gene expression across individuals. Long non-coding RNAs (lncRNAs) are likely to be important regulators within the <i>CYP3A5</i> regulatory network in the liver. In this study, we employed a computational approach to estimate Sobol's sensitivity indices (SSI) under generalized linear models, applied to liver RNA expression microarray data (GTEx v8). The SSI-based analysis revealed that the long non-coding RNA LINC02499 exhibits the highest SSI value in relation to <i>CYP3A5</i> expression in the liver. Furthermore, we conducted a comprehensive evaluation of LINC02499's biological characteristics and confirmed its regulatory role in <i>CYP3A5</i> expression through real-time quantitative PCR and Western blotting experiments at the cellular level. We also demonstrated the direct interaction between miR-329-5p and both LINC02499 and <i>CYP3A5</i> mRNA using dual-luciferase reporter assays and electrophoretic mobility shift assays (EMSA). In conclusion, our findings outline a regulatory network in which LINC02499 functions as a molecular sponge, mitigating the post-transcriptional inhibitory effect of miR-329-5p on <i>CYP3A5</i> expression.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":"99 11","pages":"4455 - 4466"},"PeriodicalIF":6.9,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144551777","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Prioritisation of co-formulants and plant protection products for non-dietary risk assessment using NAMs","authors":"Alkiviadis Stagkos-Georgiadis,&nbsp;Bright Baffour-Duah,&nbsp;Tewes Tralau,&nbsp;Denise Bloch","doi":"10.1007/s00204-025-04078-0","DOIUrl":"10.1007/s00204-025-04078-0","url":null,"abstract":"<div><p>Plant protection products (PPPs) contain one or more active substances (AS) as well as a varying number of co-formulants. Whilst AS are amongst the most data-rich and well-investigated substances in toxicology, possibly toxic co-formulants and PPPs with potentially relevant mixture effects are not considered in risk assessment. This especially applies to operators who come in contact with the undiluted product. In this study, we investigated this concern by prioritising co-formulants and PPPs for further testing using new approach methodologies (NAMs). We combined in silico predictions of co-formulant nephrotoxicity and hepatotoxicity with AS and co-formulant kinetic interaction prediction and thus identified 427 PPPs for further investigation. In a next step, six PPPs were there assessed for their comparative AS and PPP toxicity in liver cells. One product displayed more than additive effects and was thus tested for in silico predicted kinetic interactions. Whilst permeability glycoprotein (P-gp) interaction could not be confirmed, the observed effect may likely be attributed to CYP2C19 interaction. In summary, the findings highlight that the use of in silico tools as a screening step is useful to limit the number of PPPs for further testing. Subsequent in vitro testing of a limited number of PPPs supported the application of the additivity concept and highlighted that prioritised co-formulants contribute additively. We propose the integration of such co-formulants into the Hazard Index (HI) approach currently applied in active substance combined risk assessment.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":"99 8","pages":"3205 - 3221"},"PeriodicalIF":6.9,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00204-025-04078-0.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144551778","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}