Journal of Toxicological Sciences最新文献

{"title":"Neurotoxicity of acrylamide in wild-type and TNF-α depletion mice: possible alternative role of IL-6 and dipolar effects of TNF-α depletion on oxidative stress pathway.","authors":"Cai Zong, Harue Sato, Sahoko Ichihara, Yoichiro Iwakura, Seiichiroh Ohsako, Gaku Ichihara","doi":"10.2131/jts.51.183","DOIUrl":"10.2131/jts.51.183","url":null,"abstract":"<p><p>Neurotoxicity of acrylamide has been demonstrated both in humans and animals, while the mechanisms remain largely unknown. We recently reported TNF-α deletion suppressed acrylamide-induced neurotoxicity in mice at low dose. Here we further investigated expression of antioxidant and proinflammatory cytokines to explore roles of TNF-α. Wild type and TNF-α KO mice were exposed to acrylamide at 0/12.5/25 mg/kg bw for 28 days. The results showed that acrylamide significantly decreased body weight at 12.5 and 25 mg/kg bw, but decreased brain weight only at 25 mg/kg bw. TNF-α deletion didn't alleviate the above effects. Also, TNF-α deletion didn't alleviate decrease of grip strength at 25 mg/kg bw. Immunohistochemical results showed that TNF-α deletion alleviated noradrenergic axon degeneration in cortex S1FL and S1HL regions at 12.5 mg/kg bw, but not 25 mg/kg bw. Moreover, TNF-α deletion suppressed acrylamide-induced upregulation of TGF-β and NF-κB, but didn't suppress upregulation of IL-6, suggesting possible roles of IL-6 in acrylamide-induced neurotoxicity, particularly at high concentration. Moreover, this study showed a dipolar effect of TNF-α deletion on oxidative stress pathways, i.e., at 25 mg/kg bw, TNF-α deletion suppressed upregulation of oxidative stress (Keap1/HO-1/Gclc/Gclm/Sod/Cat/Gstm/MT-1); however, at 12.5 mg/kg bw, TNF-α deletion accelerated upregulation of Nqo1/Gclm/Sod1/Cat/Gsr. Taken together, genetic TNF-α ablation, at least partially, alleviated acrylamide neurotoxicity at low concentration. Limited alleviation effects at high concentration generated a hypothesis that this may be due to IL-6 signaling and dipolar regulating effects of TNF-α deletion on oxidative stress pathway. This study provided new insights into acrylamide neurotoxicity and TNF-α-targeting strategy.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"51 3","pages":"183-199"},"PeriodicalIF":1.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147326407","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of the stability and cross-reactivity of zopiclone and eszopiclone using immunoassay kits.","authors":"Takeshi Saito, Akira Namera, Rie Yamamoto, Seiji Morita, Yoshihide Nakagawa","doi":"10.2131/jts.51.173","DOIUrl":"10.2131/jts.51.173","url":null,"abstract":"<p><p>Zopiclone (ZOP) and eszopiclone (EZOP) are hypnotics belonging to the cyclopyrrolone class, which are metabolized to zopiclone-N-oxide (ZOPNO) and N-desmethylzopiclone (NDZOP), respectively, and finally to 2-amino-5-chloropyridine (2A5C) after long-term storage, freezing, and conservation. This study aimed to examine the stability of ZOP and EZOP using a commercially available immunoassay urine screening kit for ZOP. ZOP and EZOP, as well as their metabolites and 2A5C, were analyzed in urine using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Spiked urine samples were used to confirm cross-reactivity for the immunoassay test and to compare the quantitative results of the clinical urine samples. The results indicate that urine samples stored for an extended period or altered postmortem may lead to false-negative results for ZOP, providing important information for interpreting results in cases involving ZOP.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"51 3","pages":"173-181"},"PeriodicalIF":1.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147326451","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Induction of S100A11 calcium-binding protein in renal tubular epithelial cells during the formation of calcium oxalate crystals.","authors":"Shintaro Isa, Toshihiko Aki, Kanako Noritake, Atsushi Yamada, Kana Unuma","doi":"10.2131/jts.51.489","DOIUrl":"https://doi.org/10.2131/jts.51.489","url":null,"abstract":"<p><p>Acute kidney injuries and renal dysfunction caused by ethylene glycol poisoning are often associated with nephrolithiasis. Calcium oxalate (CaOx) crystal formation is considered one of the most common events at the early stage of nephrolithiasis. The aim of this study is to investigate molecular mechanisms underlying CaOx crystal formation on renal tubular epithelial cells in vitro. We applied oxalate or potassium oxalate (final concentrations, 200-1200 µM) in the medium culturing NRK-52E rat proximal tubular epithelial cells. Cell death was observed with levels of oxalate or potassium oxalate above 800 µM 48 hr after addition. Crystals, mainly consisting of CaOx as confirmed by polarized light microscopy, were detected on cells treated with 800 µM oxalate or potassium oxalate. Transcriptome analysis showed that a gene (S100A11-like) belonging to the S100 family, which consists of homologous Ca-binding proteins, many of which can form complexes with annexin family proteins, was the gene most upregulated by 800 µM oxalate. Immunoblot as well as immunocytochemical analysis indicated not only upregulation but also alteration of intracellular localization of S100A11. The same altered pattern of localization was also observed for annexin A2, which has been shown to be involved in the formation of CaOx crystals. This study presents the possibility that, in addition to annexin family proteins, S100 family proteins might be involved in the regulation of CaOx crystal formation and/or other events associated with crystal formation.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"51 9","pages":"489-500"},"PeriodicalIF":1.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148881041","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Variable hepatic and renal manifestations following grass carp gallbladder ingestion: a three-patient case series from Vietnam.","authors":"Nguyen Dang Duc, Nguyen Trung Nguyen, Lam Nguyen Hong Khanh, Le Thi Dieu Hien","doi":"10.2131/jts.51.527","DOIUrl":"https://doi.org/10.2131/jts.51.527","url":null,"abstract":"<p><p>Fish gallbladder ingestion remains a traditional folk remedy in several Asian countries but may result in severe hepatorenal toxicity. We report three patients admitted to the Poison Control Center, Bach Mai Hospital, Hanoi, Vietnam, following ingestion of grass carp (Ctenopharyngodon idella) gallbladders. A 44-year-old man developed severe hepatocellular injury (peak AST 4442 U/L, ALT 5115 U/L), moderate acute kidney injury (AKI), and prolonged severe hypertension requiring intravenous nicardipine infusion for 88 hr. A 48-year-old man presented with severe AKI (peak creatinine 749 µmol/L) and toxic hepatitis and underwent intermittent hemodialysis because of rapidly progressive renal dysfunction despite preserved urine output. A 73-year-old woman developed severe acute hepatocellular injury with AST and ALT levels exceeding 7000 U/L and ferritin of 40,000 ng/mL but without clinically significant AKI. Peak total bilirubin concentrations were 83.6, 102, and 99 µmol/L in Cases 1, 2, and 3, respectively. Viral hepatitis investigations were negative in all patients. Despite variable clinical phenotypes, all patients recovered with supportive treatment. These cases further illustrate the variable patterns of hepatorenal involvement following grass carp gallbladder ingestion, ranging from dialysis-requiring acute kidney injury to predominant hepatocellular injury with preserved renal function. Because toxin exposure could not be quantified, the observed variability may reflect differences in ingested toxin dose, host susceptibility, or both. These findings highlight interindividual variability in patterns of hepatorenal involvement following grass carp gallbladder ingestion.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"51 10","pages":"527-532"},"PeriodicalIF":1.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149111579","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Charcoal hemoperfusion for aconite poisoning: Insights from a CKD case and complementary simulation experiments.","authors":"Yoshiaki Oshima, Akihiro Otsuki, Yukari Minami, Masaharu Fukuki, Naoki Moriyama, Aki Aoki, Ryo Endo, Masato Nakasone, Yoshinori Kitagawa, Hiroyuki Minato, Shuhei Nakaji, Akira Tanaka","doi":"10.2131/jts.51.413","DOIUrl":"https://doi.org/10.2131/jts.51.413","url":null,"abstract":"<p><p>Aconite contains four highly toxic diester-diterpene alkaloids (DDAs): aconitine, mesaconitine, hypaconitine, and jesaconitine. The efficacy of charcoal hemoperfusion (CHP) in treating aconite poisoning has recently been reported in several East Asian cases. This study evaluated the potential efficacy of CHP using two simulation experiments and one clinical case. One simulation examined the saturated adsorption capacity of activated charcoal for aconitine, whereas the other assessed the time course of perfusate aconitine concentrations at the inlet and outlet of an activated-charcoal column. The clinical case involved a 61-year-old female with stage V chronic kidney disease who underwent CHP for an electrical storm due to aconite poisoning. Her initial plasma DDA concentration was markedly higher than those reported in previous cases. This elevated concentration raises the possibility of substantial ingestion, although the exact amount cannot be determined. CHP was initiated 2 hr after ingestion and continued for 7 hr. In the simulations, activated charcoal showed a saturated adsorption capacity of 110 mg of aconitine per gram, and the column removed most aconitine in a single pass. In the clinical case, plasma DDA concentrations rapidly decreased after 7 hr of CHP, with a clearance rate of approximately 70 mL/min at a blood-flow rate of 100 mL/min. The terminal elimination half-life of plasma DDAs was 109.5 hr, likely reflecting the patient's underlying kidney disease. Taken together, these findings suggest that CHP may contribute to DDA elimination under severe renal impairment, although its broader clinical relevance remains uncertain.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"51 8","pages":"413-426"},"PeriodicalIF":1.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148664072","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Possible involvement of tubular interleukin-34 in macrophage recruitment during colistin-induced nephrotoxicity in rats.","authors":"Kohei Matsushita, Hirotoshi Akane, Jun-Ichi Akagi, Tomomi Morikawa, Yasuko Mizuta, Kumiko Ogawa, Takeshi Toyoda","doi":"10.2131/jts.51.163","DOIUrl":"10.2131/jts.51.163","url":null,"abstract":"<p><p>Colistin is a cationic cyclic lipopeptide antibiotic used against multidrug-resistant Gram-negative bacteria; however, its clinical application is limited by nephrotoxicity. Although oxidative stress, mitochondrial dysfunction, and endoplasmic reticulum stress are implicated in the primary cytotoxic mechanisms, the secondary inflammatory pathways remain poorly understood. In this study, we aimed to elucidate the mechanisms underlying colistin-induced nephrotoxicity by analyzing the molecular responses of injured renal tubules isolated by laser microdissection (LMD) in a rat model. Six-week-old male Sprague-Dawley rats were subcutaneously administered colistin (0, 15, or 30 mg/kg/day) for 28 days. Increases in serum creatinine and blood urea nitrogen were associated with tubular vacuolation, single-cell necrosis, and regenerative changes in proximal tubules. Immunohistochemistry revealed increased expression of kidney injury molecule-1 (KIM-1) and presence of cleaved caspase-3 in the injured tubules, indicating epithelial regeneration and apoptosis, respectively. LMD-based microarray analysis identified 486 upregulated and 472 downregulated genes in vacuolated/regenerative tubules compared with normal tubules. Pathway analysis indicated activation of immune-related processes, particularly associated with macrophage activation and trafficking, including interleukin-34 (IL-34). In situ hybridization confirmed Il34 mRNA expression in the cytoplasm of the injured tubules, and accumulation of CD68-positive macrophages around KIM-1-positive tubules. Conversely, no significant change was observed in CD163-positive macrophages following colistin treatment, suggesting proinflammatory M1 macrophage predominance. These findings indicate that tubular IL-34 induction and subsequent macrophage recruitment amplify colistin-induced nephrotoxicity at the secondary level, suggesting that proximal tubules function as both primary targets and effectors of inflammation. Targeting the IL-34-related signaling could thus serve as a potential approach to alleviate colistin-induced renal injury.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"51 3","pages":"163-172"},"PeriodicalIF":1.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147326391","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of zwitterionic liquids on cytochrome P450 and P-glycoprotein.","authors":"Yusuke Kubota, Risa Funaki, Yuto Sana, Takeru Ishizaki, Richard W Wong, Eishu Hirata, Kosuke Kuroda","doi":"10.2131/jts.51.303","DOIUrl":"10.2131/jts.51.303","url":null,"abstract":"<p><p>OE₂imC₃C, a zwitterionic ionic liquid, has attracted considerable attention as a low-toxicity solvent. However, the effects of OE₂imC₃C on the pharmacokinetics of drugs have not yet been investigated. In this study, we examined the effects of OE₂imC₃C on p-glycoprotein (P-gp) and cytochrome P450 (CYPs), typical factors associated with pharmacokinetic interactions. No notable inhibition of P-gp by OE₂imC₃C was observed at the tested concentrations. OE₂imC₃C inhibited several CYPs (IC₅₀:0.048-0.66 vol% for 70 wt% aq OE₂imC₃C stock solution) and induced the gene expression of several CYPs. Based on these findings, practical guidance for the use of OE₂imC₃C was proposed here. A previous study suggested that OE₂imC₃C does not penetrate the cell membrane, and therefore, we had hypothesized that it does not inhibit CYPs inside cells. However, OE₂imC₃C inhibited CYPs. The related transporter was assumed to be organic cation transporter 1 (OCT1). These results suggest that OE₂imC₃C effect on CYPs should be considered in both in vitro and in vivo experiments. The CYP inhibition was alleviated through structural modification of OE₂imC₃C.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"51 5","pages":"303-314"},"PeriodicalIF":1.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147839777","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Caffeine potentiates the dependence induced by central nervous system depressants contained in over-the-counter medications in mice.","authors":"Ayaka Yonemura-Hirata, Asuka Kaizaki-Mitsumoto, Satoshi Numazawa","doi":"10.2131/jts.51.131","DOIUrl":"10.2131/jts.51.131","url":null,"abstract":"<p><p>Over-the-counter (OTC) medicines are available without a prescription and are key to the promotion of self-medication. However, they also pose the risks of misuse, overdose, addiction, and abuse. These risks have recently emerged as global public health concerns. An important aspect of OTC drugs in Japan is that they are often combined with drugs with different effects. Although it has been noted that interactions between depressants and stimulants of the central nervous system (CNS) may promote drug dependence, the details remain unclear due to a lack of basic evidence. Therefore, an assessment was conducted to determine the interaction between CNS depressants, including dextromethorphan, diphenhydramine, and bromovalerylurea, and the CNS stimulant caffeine, by employing a conditioned place preference test in mice. Even at low doses, long-term administration of dextromethorphan and diphenhydramine induced place preference. Long-term administration of high-dose bromovalerylurea also induced this effect. The period required for dextromethorphan, diphenhydramine, bromovalerylurea, and morphine to acquire place preference was shortened by co-administration with caffeine, demonstrating that CNS stimulation enhances the preference of these sedatives in mice. Moreover, the preference for these drugs was suppressed by the dopamine D₁ receptor antagonist SCH23390, and by the dopamine D₂ receptor antagonist sulpiride, suggesting that dopamine is involved in the enhancing effect. These findings underscore the need to reconsider the active ingredients and distribution practices of OTC products, as the prolonged or inappropriate use of OTC medications and polypharmacy increases the risk of dependence.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"51 2","pages":"131-139"},"PeriodicalIF":1.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146100330","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative cytotoxicity of novel mercury species α-mercuri-acetaldehyde and α-mercuri-acetic acid versus methylmercury in SH-SY5Y cells.","authors":"Kaito Yamashiro, Shun Kono, Takumi Katsuzawa, Sachie Arae, Ryo Irie, Yuuki Fujimoto, Tsutomu Takahashi, Yasuyuki Fujiwara, Yasuhiro Shinkai, Toshiyuki Kaji, Yo Shinoda","doi":"10.2131/jts.51.141","DOIUrl":"10.2131/jts.51.141","url":null,"abstract":"<p><p>Recently, α-mercuri-acetaldehyde (HgCH₂CHO) and α-mercuri-acetic acid (HgCH₂COOH) have been proposed as potential causative agents of Minamata disease. However, their toxicological profiles remain largely unknown. This study aimed to characterize the cytotoxicity, cellular uptake, and efflux mechanisms of these compounds in SH-SY5Y neuroblastoma cells and to compare these properties with those of methylmercury (MeHg). Cell viability was assessed after 24 hr of exposure to MeHg (1-10 µM), HgCH₂CHO (10-50 µM), or HgCH₂COOH (10-50 µM) using the CCK-8 assay. The roles of L-type amino acid transporter 1 (LAT1) and multidrug resistance-associated proteins (MRPs) were evaluated using the inhibitors JPH203 (1 µM) and MK571 (10 µM), respectively. Intracellular mercury accumulation was quantified after 24 hr of exposure to 3 µM of each compound using thermal decomposition-amalgamation atomic absorption spectrometry. All compounds exhibited dose-dependent cytotoxicity, with a relative toxicity order of MeHg (LC₅₀: 6.4 µM) > HgCH₂CHO (LC₅₀: 14.6 µM) > HgCH₂COOH (LC₅₀: 39.2 µM). LAT1 inhibition had minimal effect on MeHg toxicity but slightly attenuated that of HgCH₂CHO and HgCH₂COOH. Conversely, MRP inhibition markedly enhanced MeHg toxicity, modestly increased that of HgCH₂CHO, and slightly increased that of HgCH₂COOH. Cellular mercury accumulation was consistent with cytotoxicity patterns, showing 10-20-fold lower levels for HgCH₂CHO and HgCH₂COOH than for MeHg. HgCH₂CHO and HgCH₂COOH were approximately 2-5-fold less cytotoxic than MeHg and exhibited substantially lower intracellular mercury levels. Our findings suggest that HgCH₂CHO and HgCH₂COOH are unlikely to have neurotoxic potential comparable to that of MeHg.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"51 2","pages":"141-147"},"PeriodicalIF":1.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146100324","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Elemental impurities in Yataprasen Thai Polyherbal Formulation: source-dependent variation, and analytical method validation.","authors":"Jaenjira Angsusing, Yu-Ki Tanaka, Chuda Chittasupho, Yasumitsu Ogra","doi":"10.2131/jts.51.111","DOIUrl":"10.2131/jts.51.111","url":null,"abstract":"<p><p>The global consumption of complementary and alternative medicine (CAM) has risen dramatically. However, safety concerns persist owing to contamination with elemental impurities. In this study, we optimized and validated an analytical method for quantifying four toxic metals, As, Cd, Pb, and Hg, in Yataprasen (YTPS) extract, a complex Thai traditional polyherbal formulation. Samples were obtained from natural collection sites and commercial sources, and the contributions of 13 individual herbs to total impurities were evaluated. Inductively coupled plasma mass spectrometry (ICP-MS) was applied following four acid-digestion protocols. Quantification involved external calibration and standard addition, with validation covering LOD, LOQ, linearity, precision, and recovery. Toxicological risk was assessed in accordance with the ICH Q3D(R2) guideline. Digestion with 1 mL of HNO₃ gave the highest accuracy and recovery. Cd and Pb levels showed little variation across methods. Validation demonstrated excellent accuracy (93.6-107.5% recovery), strong linearity (R² > 0.998), and low detection limits (<1.5 µg/kg). A significant difference in elemental impurity concentration was observed between the two sources, with the naturally collected YTPS exhibiting markedly higher levels of all four metals than the commercial one. While external calibration was sufficient for commercial samples, standard addition was required for naturally sourced samples to overcome matrix effects. Component analysis identified Allium sativum L., Cymbopogon nardus (L.) Rendle, and Melia azedarach L. as the major contributors to the impurity burden. Source-dependent variation in elemental impurity concentration was observed in YTPS extracts, with natural collection posing a greater toxicological concern. The validated analytical workflow provides a robust platform for quality control and regulatory assessment of traditional polyherbal formulations.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"51 2","pages":"111-122"},"PeriodicalIF":1.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146100367","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}