Derivation of a chronic reference dose and reference concentration for trimethylbenzenes and C9 aromatic hydrocarbon solvents

Trimethylbenzenes (TMBs) and C9 aromatic hydrocarbon solvents are structurally similar and have similar toxicity. Based on a review of the entire TMB and C9 aromatic hydrocarbon solvents toxicology database, oral and inhalation studies were identified to serve as the basis for a Reference dose (RfD) and Reference concentrations (RfC). The RfD and RfC were derived using standard USEPA methods and assumptions. The RfD was calculated to be 0.4 mg/kg/day using a 90-day oral study that resulted in a NOAEL of 600 mg/kg/day, based on a lack of adverse effects at the highest dose level (reversible effects such as increased serum phosphorus levels and liver and kidney weights), along with a total uncertainty factor of 1000. For the RfC, three studies were considered based on different study designs and toxicological endpoints, including neurotoxicity, systemic toxicity, and potential developmental and reproductive toxicity. For all three studies, as the calculated RfCs were consistent (3–4 mg/m³), the most conservative RfC, 3 mg/m³, was selected. The C9 aromatic hydrocarbon solvents referred to herein are based on chemistries assessed as part of the TSCA Section 4 Test Rule. These solvents contain primarily ethyl toluene and tri-methyl benzene isomers, but the specific compositions can vary based on feedstock and manufacturing process, thus, it is important to consider the composition of any specific solvent to assess similarity to that assessed in the TSCA Section 4 Test Rule program.     

Proposal for a revised Reference Concentration (RfC) for manganese based on recent epidemiological studies

In 1993, based on observations of subclinical neurological effects in workers, the United States Environmental Protection Agency (US EPA) published a Reference Concentration (RfC) of 0.05 μg/m³ for manganese (Mn). The geometric mean exposure concentration, 150 μg/m³ respirable Mn, was considered the lowest observable adverse effect level (LOAEL), and uncertainty factors (UFs) were applied to account for sensitive populations, database limitations, a LOAEL, subchronic exposure, and potential differences in toxicity of different forms of Mn. Based on a review of more recent literature, we propose two alternate Mn RfCs. Of 12 more recent occupational studies of eight cohorts with chronic exposure durations, examining subclinical neurobehavioral effects, predominantly on the motor system, three were considered appropriate for development of an RfC. All three studies yielded no observable adverse effect levels (NOAELs) of approximately 60 μg/m³ respirable Mn. Converting the occupational NOAEL to a human equivalent concentration (HEC) of 21 μg/m³ (for continuous exposure) and applying a UF of 10 to account for intraspecies variability yielded an RfC of 2 μg/m³. We also derived a similar RfC (7 μg/m³) using an Mn benchmark dose (BMD) as the point of departure. Overall confidence in both RfCs is medium.     

Derived Reference Doses (RfDs) for the environmental degradates of the herbicides alachlor and acetochlor: Results of an independent expert panel deliberation

An independent peer expert panel was convened under the auspices of the Alliance for Risk Assessment (ARA) to review toxicology data and derive oral Reference Doses (RfDs) for four environmental degradates of the acetanilide herbicides, alachlor and acetochlor. The degradates included in this evaluation were (1) alachlor tertiary-ethanesulfonic acid (ESA), (2) alachlor tertiary-oxanilic acid (OXA), (3) acetochlor ESA, and (4) acetochlor OXA. Each degradate was judged to have sufficient data for developing low to medium confidence RfD, with use of an additional uncertainty factor (UF) to cover data gaps. Body weight decreases were identified as the most sensitive treatment-related adverse effect for RfD development. A composite UF of 1000 (10 for human variability in sensitivity, 10 for interspecies differences in sensitivity, and 10 for subchronic to chronic and database deficiency combined; i.e., 10_A × 10_H × 10_S&D) for each degradate was considered reasonable, while noting that an argument could be made for an UF of 3000 (10_A × 10_H × 30_S&D). Based on the available data, an oral RfD of 0.2 mg/kg-day is recommended for both acetochlor ESA and acetochlor OXA and an oral RfD of 0.8 mg/kg-day is recommended for both alachlor ESA and alachlor OXA.     

Derivation of a toxicity reference value for nitrogen trichloride as a disinfection by-product

Nitrogen trichloride is a highly volatile chlorination disinfection by-product, very commonly found in the air of indoor swimming pools. The aim of this work is to characterize the hazard associated with it and to determine the concentration at which health effects appear, for application in health risk assessments for users of indoor swimming pools. Hazard identification was based on a literature survey and analysis of animal and human studies, with special attention paid to their methodological quality and to reports of a dose–response relationship. A toxicity reference value was derived for respiratory effects, based on human data from both general and occupational data. We selected a lowest-observed-adverse-effect-level of 0.355 mg/m³ based on objective measurements rather than self-reported effects. Two uncertainty factors were applied to take into account both intra-species variability and the use of a concentration with an effect rather than a no-observed-adverse-effect-level. A toxicity reference value of 4 × 10^?3 mg/m³ for nitrogen trichloride is proposed for repeated short exposures. Alternative values based on animal data range from 0.01 to 0.03 mg/m³.     

Comparison of PBTK model and biomarker based estimates of the internal dosimetry of acrylamide

Estimates of internal dosimetry for acrylamide (AA, 2-propenamide; CASRN: 79-06-1) and its active metabolite glycidamide (GA) were compared using either biomarkers of internal exposure (hemoglobin adduct levels in rats and humans) or a PBTK model (Sweeney et al., 2010). The resulting impact on the human equivalent dose (HED, oral exposures), the human equivalent concentration (HEC, inhalation), and final reference values was also evaluated. Both approaches yielded similar AA HEDs and HECs for the most sensitive noncancer effect of neurotoxicity, identical oral reference doses (RfD) of 2 × 10⁻³ mg AA/kg bw/d, and nearly identical inhalation reference concentrations (RfC = 0.006 mg/m³ and 0.007 mg/m³, biomarker and PBTK results, respectively). HED and HEC values for carcinogenic potential were very similar, resulting in identical inhalation unit risks of 0.1/(mg AA/m³), and nearly identical oral cancer slope factors (0.4 and 0.5/mg AA/kg bw/d), biomarker and PBTK results, respectively. The concordance in estimated HEDs, HECs, and reference values from these two diverse methods increases confidence in those values. Advantages and specific application of each approach are discussed.(Note: Reference values derived with the PBPK model were part of this research, and do not replace values currently posted on IRIS: http://www.epa.gov/iris/toxreviews/0286tr.pdf.)     

Derivation of a chronic oral reference dose for cobalt

Cobalt (Co) is an essential element in humans as a component of vitamin B12. However, at high levels Co exposure has been shown to have detrimental effects. This study was designed to identify a chronic oral reference dose (RfD) for Co. Currently available data indicate that non-cancer health effects associated with Co exposure may include hematological, neurological, immunological, reproductive, cardiovascular, and endocrine responses. This analysis employs the standard US EPA risk assessment methodology for establishing a chronic RfD. In this analysis, the Jaimet and Thode (1955) 10-week, multiple dose human study of thyroid effects (decreased iodine uptake) in children was determined to be the most robust and sensitive study for identifying a potential point of departure dose (POD). A dose of 0.9 mg Co/kg-day was chosen as the POD. Consistent with the US EPA’s previous derivation of the perchlorate RfD, which is also based on decreased iodine uptake in humans, we considered several uncertainly factors (UFs), and determined that a factor of 10 for human variability was appropriate, as well as a factor of three for database adequacy. Applying an aggregate uncertainty factor of 30 to the POD yields a chronic oral RfD of 0.03 mg/kg-day. We believe this value would be protective of non-cancer health effects in the general population for a lifetime of daily exposure to Co.     

Mode of action analysis for liver tumors from oral 1,4-dioxane exposures and evidence-based dose response assessment

1,4-Dioxane is found in consumer products and is used as a solvent in manufacturing. Studies in rodents show liver tumors to be consistently reported after chronic oral exposure. However, there were differences in the reporting of non-neoplastic lesions in the livers of rats and mice. In order to clarify these differences, a reread of mouse liver slides from the 1978 NCI bioassay on 1,4-dioxane in drinking water was conducted. This reread clearly identified dose-related non-neoplastic changes in the liver; specifically, a dose-related increase in the hypertrophic response of hepatocytes, followed by necrosis, inflammation and hyperplastic hepatocellular foci. 1,4-Dioxane does not cause point mutations, DNA repair, or initiation. However, it appears to promote tumors and stimulate DNA synthesis. Using EPA Guidelines (2005), the weight of the evidence suggests that 1,4-dioxane causes liver tumors in rats and mice through cytotoxicity followed by regenerative hyperplasia. Specific key events in this mode of action are identified. A Reference Dose (RfD) of 0.05 mg/kg day is proposed to protect against regenerative liver hyperplasia based on a benchmark dose (BMD) approach. Based on this RfD, a maximum contaminant level goal of 350 μg/L is proposed using a default relative source contribution for water of 20%.     

Application of a physiologically based pharmacokinetic model for reference dose and reference concentration estimation for acetone

Recent health risk assessments to propose a Reference Dose (RfD) for acetone (Forsyth, 2001; U.S. EPA, 2001) have been based on the results of an oral subchronic study conducted in rats and mice (Dietz et al., 1991; NTP, 1991). These assessments have utilized the traditional concept of establishing the RfD by determining the lowest experimentally determined No-Observed-Adverse-Effect Level (NOAEL) and applying various Uncertainty Factors (UFs) (U.S. EPA, 1988). This article describes a risk assessment for acetone based on the systemic toxicity observed in subchronic and developmental toxicity studies to estimate an RfD and an inhalation reference concentration (RfC) for acetone. Specifically, this approach examined the subchronic study by Dietz et al. (1991), as well as an inhalation developmental toxicity study on acetone (Mast et al., 1988) and several toxicology studies of isopropanol (IPA). This was accomplished by applying a physiologically based pharmacokinetic (PBPK) model developed previously for IPA and its metabolite acetone (Clewell et al., 2001). The incorporation of the PBPK model into the derivation of an RfD and RfC for acetone allowed for a tissue-based approach rather than an external exposure-based approach, making it possible to derive an oral RfD from an inhalation study. In addition, the use of the PBPK model to analyze data from chronic and reproductive/developmental studies conducted with IPA enabled an assessment of the potential for acetone to produce any of the effects observed in the IPA studies. This analysis provided sufficient information to reduce the need for UFs in the adjustment of the NOAEL from the oral subchronic study for the determination of an RfD. Using the PBPK model in the acetone risk assessment supports a composite UF of 60 for the subchronic study, compared to composite factors of 300 to 3000 in the other recent risk assessments. This difference resulted in an RfD of 16 mg/kg/d, compared to the values of 0.3 to 3 that have previously been estimated (Forsyth, 2001; U.S. EPA, 2001). Considering the results from the inhalation developmental study (Mast et al., 1988) resulted in an RfD of 8.7 mg/kg/d. Using this study also fills a data gap for acetone that exists if only the oral database for acetone is considered for RfD derivation. An RfC of 29 ppm was also estimated for acetone using the Mast et al. (1988) study results in combination with the PBPK model. The potential impact of endogenous acetone on a risk assessment for acetone is also discussed.     

An antioxidant,N,N′-diphenyl-p-phenylenediamine (DPPD), affects labor and delivery in rats: A 28-day repeated dose test and reproduction/developmental toxicity test

A 28-day repeated dose toxicity test and reproduction/developmental toxicity test for N,N′-diphenyl-p-phenylenediamine (DPPD) were conducted in [Crl:CD(SD)] SPF rats. Male and female rats were dosed with DPPD by gavage for 28 days at 0, 100, 300, or 1000 mg/kg bw/day or for a total of 42–46 days at 0, 8, 50, or 300 mg/kg bw/day. No significant adverse effects were observed in the repeated dose toxicity study up to 1000 mg/kg bw/day in both sexes. In the reproduction/developmental toxicity study, two females showed piloerection, hypothermia, and pale skin; one died and the other showed dystocia on day 23 of pregnancy at 300 mg/kg bw/day. Another female delivered only three live pups at 300 mg/kg bw/day. A significantly prolonged gestation period was observed at 50 and 300 mg/kg bw/day. The NOAELs of repeated dose toxicity and reproduction/developmental toxicity were considered to be 1000 and 8 mg/kg bw/day, respectively.     

Effect of styrene exposure on plasma parameters,molecular mechanisms and gene expression in rat model islet cells

Styrene is an aromatic hydrocarbon compound present in the environment and have primary exposure through plastic industry. The current study was designed to evaluate styrene-induced toxicity parameters in rat plasma fasting blood glucose (FBG) level, oral glucose tolerance, insulin secretion, oxidative stress, and inflammatory cytokines in cellular and molecular levels. Styrene was dissolved in corn oil and administered at different doses (250, 500, 1000, 1500, 2000 mg/kg/day and control) to each rat, for 42 days. In treated groups, styrene significantly increased fasting blood glucose, plasma insulin (p < 0.001) and glucose tolerance. Glucose tolerance, insulin resistance and hyperglycemia were found to be the main consequences correlating gene expression of islet cells. Styrene caused a significant enhancement of oxidative stress markers (p < 0.001) and inflammatory cytokines in a dose and concentration-dependent manner in plasma (p < 0.001). Moreover, the activities of caspase-3 and −9 of the islet cells were significantly up-regulated by this compound at 1500 and 2000 mg/kg/day styrene administrated groups (p < 0.001). The relative fold change of GLUD1 was downregulated (p < 0.05) and upregulated at 1500 and 2000 mg/kg, respectively (p < 0.01). The relative fold changes of GLUT2 were down regulated at 250 and 1000 mg/kg and up regulated in 500, 1500 and 2000 mg/kg doses of styrene (p < 0.01). The expression level of GCK indicated a significant upregulation at 250 mg/kg and downregulation of relative fold changes in the remaining doses of styrene, except for no change at 2000 mg/kg of styrene for GCK. Targeting genes (GLUD1, GLUT2 and GCK) of the pancreatic islet cells in styrene exposed groups, disrupted gluconeogenesis, glycogenolysis pathways and insulin secretory functions. The present study illustrated that fasting blood glucose, insulin pathway, oxidative balance, inflammatory cytokines, cell viability and responsible genes of glucose metabolism are susceptible to styrene, which consequently lead to other abnormalities in various organs.     

Application of a physiologically based pharmacokinetic model for isopropanol in the derivation of a reference dose and reference concentration

An interspecies physiologically based pharmacokinetic (PBPK) model describing isopropanol (IPA) and its major metabolite, acetone, was applied to perform route-to-route and cross-species dosimetry to derive reference dose (RfD) and reference concentration (RfC) values for IPA. Adult PBPK models for rats and humans were extended to simulate exposure to IPA during pregnancy and used to estimate internal dose metrics in the mother and fetus during development. Endpoints from chronic, developmental, and reproductive toxicity studies were considered for the derivation of RfDs and RfCs. Due to uncertainties in the mode of action of toxicity for IPA and acetone, the dose metric used for most responses was the total area under the blood concentration curve (AUC) for the combination of IPA and acetone. This combined dose metric provided a more conservative estimate than those based on AUCs for IPA or acetone. Peak blood concentration of IPA was the dose metric for neurobehavioral effects. The recommended RfD and RfC for IPA are 10 mg/kg/day and 40 ppm, respectively, based on decreased fetal body weights. All of the PBPK-derived RfD or RfC values for various endpoints were similar (within a factor of 3), regardless of route of exposure in the animal study.     

Acute and sub-chronic toxicity studies of the extract of Thunberg Fritillary Bulb

The aim of this study was to investigate the acute and sub-chronic toxicity of extract of Thunberg Fritillary Bulb. For the acute toxicity tests, graded doses of the extract were administered orally to mice. The animals were observed for toxic symptoms and mortality daily for 14 days. In the sub-chronic toxicity study, rats were orally administered the extract at doses of 1 and 3 mg/kg body weight (BW) for 26 weeks. After 26 weeks, the rats were sacrificed for hematological, biochemical and histological examination. In the acute toxicity tests, the estimated median lethal dosage (LD₅₀) was 52.2 mg/kg body weight in the mice. In the sub-chronic toxicity tests, a dose of 1 mg/kg body weight presented no toxicity. Above the 1 mg/kg dose, the main adverse signs observed in male rats were body or head tremor and spontaneous motor activity reduction. There were no other significant changes observed in hematology, blood biochemistry, organ weight and organ histology. The overall findings of this study indicate that the extract of Thunberg Fritillary Bulb is non-toxic up to 1 mg/kg body weight, which can be considered a safe application dose.     

Derivation of an occupational exposure limit for inorganic borates using a weight of evidence approach

Inorganic borates are encountered in many settings worldwide, spurring international efforts to develop exposure guidance (US EPA, 2004; WHO, 2009; ATSDR, 2010) and occupational exposure limits (OEL) (ACGIH, 2005; MAK, 2011). We derived an updated OEL to reflect new data and current international risk assessment frameworks. We assessed toxicity and epidemiology data on inorganic borates to identify relevant adverse effects. International risk assessment frameworks (IPCS, 2005, 2007) were used to evaluate endpoint candidates: reproductive toxicity, developmental toxicity, and sensory irritation. For each endpoint, a preliminary OEL was derived and adjusted based on consideration of toxicokinetics, toxicodynamics, and other uncertainties. Selection of the endpoint point of departures (PODs) is supported by dose–response modeling. Developmental toxicity was the most sensitive systemic effect. An OEL of 1.6 mg B/m³ was estimated for this effect based on a POD of 63 mg B/m³ with an uncertainty factor (UF) of 40. Sensory irritation was considered to be the most sensitive effect for the portal of entry. An OEL of 1.4 mg B/m³ was estimated for this effect based on the identified POD and an UF of 1. An OEL of 1.4 mg B/m³ as an 8-h time-weighted average (TWA) is recommended.     

Characterisation and toxicological assessment of Neutral Methacrylate Copolymer for GRAS evaluation

Neutral Methacrylate Copolymer is a fully polymerised copolymer used in the pharmaceutical industry to permit pH-independent delayed release of active ingredients from oral dosage forms. This function has potential use with food supplements and this article describes available information on the safety of the substance.Oral administration of radiolabelled copolymer to rats resulted in the detection of chemically unchanged copolymer in the faeces, with negligible absorption. Safety studies revealed no adverse toxicity following repeated administration at doses of up to 2000 mg/kg bw/d in a sub-chronic study in rats or 250 mg/kg bw/d in a sub-chronic study in dogs. No reproductive toxicity occurred at up to 2000 mg/kg bw/d in rats or rabbits. The substance shows no evidence of genotoxicity, has low acute toxicity and no irritation or sensitisation potential.An ADI value of 20 mg/kg bw was concluded from two alternative approaches. Daily exposure from use in dietary supplements is estimated as up to 10.0 mg/kg bw in adults and 13.3 mg/kg bw in children. There would therefore appear to be no safety concerns under the intended conditions of use. The information provided is intended to support an evaluation that the substance may be “generally recognized as safe” (GRAS).     

Nitric oxide mediated effects on reproductive toxicity caused by carbon disulfide in male rats

This study investigated nitric oxide (NO) mediation of carbon disulfide (CS₂) toxicity that compromised male rat spermatogenesis and endocrine function. Rats were exposed to multiple levels of CS₂ concentration (0, 50, 250, 1250 mg/m³). A 1250 mg/m³ CS₂ + sodium nitroprusside (SNP) group and a 1250 mg/m³ CS₂ + NG-monomethyl-l-arginine (l-NMMA) group were established to explore the role of NO in mediating CS₂ toxicity. NO concentrations, NO synthase (NOS) activity, and sex hormone levels were measured, and sperm characteristics were observed and analyzed. Our data show that CS₂ exposure decreased: NOS activity; tissue NO concentrations; serum levels of gonadotropin-releasing hormones, luteinizing hormones, and testosterone; and sperm count and activity. In contrast, increased serum follicle-stimulating hormone concentrations and teratospermia were observed with CS₂ exposure. SNP reduced some of the toxic effects of CS₂, while l-NMMA treatment showed no effect. The results suggests that NO mediates compromised reproductive system function caused by CS₂ exposure.     

Reproductive and developmental toxicity screening test of 3-cyanopyridine in rats

Crl:CD(SD)rats were given 3-cyanopyridine by gavage at 0, 5, 30 or 180 mg/kg/day. Males were dosed for 42 days beginning 14 days before mating, and females for 40–53 days beginning 14 days before mating to day 3 of lactation, including throughout the mating and gestation periods. General toxicity, mainly liver damage, was observed in males at ≥30 mg/kg/day and in females at ≥5 mg/kg/day. Sertoli cell vacuolation was observed at 180 mg/kg/day, and spermatocyte damages were observed at ≥30 mg/kg/day. Effects on estrous cycles, corpora lutea and implantations, and unsuccessfully mated females, despite additional mating, were observed at 180 mg/kg/day. Delayed initiation of delivery, dystocia, and deaths or moribundities of pregnant females were observed at 180 mg/kg/day, and only two pregnant rats delivered live pups at that dose. The NOAEL for reproductive/developmental toxicity was concluded to be 30 mg/kg/day.     

Combined effects of oxytetracycline and Pb on earthworm Eisenia fetida

Combined effects of oxytetracycline (OTC) and Pb on lysosomal membrane stability and coelomocyte apoptosis of earthworm were studied in the paper. Compared with control, the lysosomal membrane stability decreased and coelomocyte apoptosis increased in the treatments of single OTC and Pb contamination. As for compound pollution, combined effect of (5 mg/kg OTC + 50 mg/kg Pb) treatment on earthworm lysosomal was synergistic (except 28 d). However, it was antagonistic at higher concentration of (10 mg/kg OTC + 50 mg/kg Pb) and (20 mg/kg OTC + 50 mg/kg Pb) treatment. In addition, coelomocyte apoptosis of earthworm decreased significantly compared with single OTC, indicating an antagonistic reaction. And joint toxicity of OTC and Pb decreased significantly with the increasing OTC concentration.     

Chemical state of heterocyclic aromatic amines in grilled beef: Evaluation by in vitro digestion model and comparison of alkaline hydrolysis and organic solvent for extraction

During grilling of the roast beef the following heterocyclic aromatic amines were found: IQ = 200.6 ng 100 g⁻¹, MeIQx = 719.8 ng 100 g⁻¹, MeIQ = 532.9 ng 100 g⁻¹, 4.8-diMeIQx = 755.4 ng 100 g⁻¹, norharmane = 507.0 ng 100 g⁻¹, harmane = 1952.6 ng 100 g⁻¹, Phe-P 1 = 263.7 ng 100 g⁻¹, Trp-P 2 = 559.2 ng 100 g⁻¹, PhIP = 1179.8 ng 100 g⁻¹ and AαC = 51.7 ng 100 g⁻¹. Their content was tested by using the method based on alkaline hydrolysis of the sample and the method based on solvent extraction of the grilled meat samples at different temperatures (without hydrolysis). The study showed that the heterocyclic aromatic amines produced during the grilling of beef are in a free form and chemically or physico-chemically bonded. The chemical forms of HAA formed in food have never been studied. For the purpose of the partial confirmation that HAA may be chemically or physico-chemically bonded, grilled beef samples were digested in vitro in model segments of the human digestive tract. Digestive enzymes, particularly proteolytic enzymes caused a statistically significant increase of free HAA determined by using solvent extraction without prior chemical hydrolysis of the sample.     

Tolerability and toxicological profile of pixantrone (Pixuvri®) in juvenile mice. Comparative study with doxorubicin

The tolerability of pixantrone dimaleate (Pixuvri^®), an aza-anthracenedione for non-Hodgkin lymphoma, was assessed in juvenile mice after intraperitoneal injection.Twenty animals/sex/dose received pixantrone 15 or 27 mg/kg/day on Post-Natal-Days (PND) 10, 13, 17, 20, 35, 39 and 42 in comparison with doxorubicin, 3 mg/kg/day. Animals were sacrificed on PND 42, 73 and 96.All pixantrone animals survived, while doxorubicin induced 52.5% mortality and the surviving animals were sacrificed early due to severe toxicity. Recoverable bone marrow toxicity (pixantrone), and toxicity to thymus and reproductive organs (pixantrone, doxorubicin) were observed without nephro- or hepatotoxicity. Pixantrone was measurable in plasma up to 2 h (occasionally 6 h) post-dose. At PND 42, mean C_max and AUC values increased proportionally with dose, without gender difference or accumulation.Pixantrone showed minimal cardiotoxicity in males and negligible in females at PND 96. Doxorubicin induced significant heart weight reduction at PND 42, however early sacrifice impeded further cardiac assessments.     

Toxicological,toxicokinetic and gastroprotective evaluation of the benzaldehyde semicarbazone

Benzaldehyde semicarbazone (BS) has presented positive results in several pharmacological models, including anticonvulsivant and anti-inflammatory models. The present study evaluated the preclinical toxicity (acute and subchronic), as well as the toxicokinetic and gastroprotective effects of BS against ethanol lesions. Oral doses of 300 and 2000 mg/kg were used in the preclinical acute toxicity study; 100, 200, and 300 mg/kg were used in both the subchronic toxicity evaluation and the gastric study; and 300 mg/kg was used in the toxicokinetic study. No impact from the dose of 300 mg/kg could be identified; while, one animal died at 2000 mg/kg in the acute toxicity test. In the subchronic toxicity test, changes in the biochemical parameters of the liver, as well as in the histopatological evaluation, demonstrated that BS is a hepatotoxic drug. BS proved to be effective for moderate and severe gastric lesions. In the toxicokinetics study, BS presented a low concentration and rapid plasma disappearance. Several results also indicate that BS is likely to be mostly eliminated from the liver and may well undergo a first-pass effect after oral absorption. It was impossible to estimate the noobserved-adverse-effect-levels (NOAEL) and lowest-observed-adverse-effect-levels (LOAEL) due to the presence of hepatotoxicity in all tested doses.