Genotoxicity Assessment Research Articles

Mutagenicity and Genotoxicity Assessment of Leuconostoc lactis DMLL10 Isolated from Kimchi.

Leuconostoc lactis DMLL10 is a microorganism specific to kimchi fermentation. In this study, we sought to evaluate the toxicity of this strain, which was newly isolated from kimchi, to determine its safety as a food ingredient. Bacterial reverse mutation assay, chromosomal aberration assay, and mammalian cell in vitro micronucleus assay were performed to assess the genetic toxicity of Leu. lactis DMLL10. The strain did not induce mutagenicity in Salmonella typhimurium TA98, TA100, TA1535, TA1537, or Escherichia coli WP2uvrA, with or without metabolic activation of S9 mixture. The oral administration of Leu. lactis DMLL10 also did not significantly increase the number of micronucleated polychromatic erythrocytes, or the mean ratio of polychromatic to total erythrocytes. Additionally, Leu. lactis DMLL10 did not cause a significant chromosomal aberration in CHU/IL cells in the presence or absence of S9 activation. Therefore, Leu. lactis DMLL10 can be suggested as a functional food ingredient with reliability and safety.

Mouthwash Containing Plant-Derived Biosurfactant and Chitosan Hydrochloride: Assessment of Antimicrobial Activity, Antibiofilm Activity, and Genotoxicity

Mouthwash Containing Plant-Derived Biosurfactant and Chitosan Hydrochloride: Assessment of Antimicrobial Activity, Antibiofilm Activity, and Genotoxicity

Improving the Assessment of Risk Factors Relevant to Potential Carcinogenicity of Gene Therapies: A Consensus Article.

Regulators and industry are actively seeking improvements and alternatives to current models and approaches to evaluate potential carcinogenicity of gene therapies (GTs). A meeting of invited experts was organized by NC3Rs/UKEMS (London, March 2023) to discuss this topic. This article describes the consensus reached among delegates on the definition of vector genotoxicity, sources of uncertainty, suitable toxicological endpoints for genotoxic assessment of GTs, and future research needs. The collected recommendations should inform the further development of regulatory guidelines for the nonclinical toxicological assessment of GT products.

Behavioural changes, DNA damage and histological alterations in Labeo rohita fingerlings in response to organic-coated silver nanoparticles.

Silver nanoparticles (AgNPs) have garnered significant global attention from researchers due to their unique physicochemical properties and wide-ranging applications in industry and medicine. However, their release into aquatic ecosystems has raised concerns regarding potential ecotoxicological consequences. The present study investigated the effects of polyvinyl pyrrolidone-coated silver nanoparticles on Labeo rohita fingerlings, focusing on behavioural reactions, genotoxic effects, histological changes and bioaccumulation. L. rohita fingerlings were exposed to polyvinyl pyrrolidone-coated silver nanoparticles with sizes ranging from 18 to 29nm for 7days at concentrations of 100, 200, 400 and 800 ug/l. The nanoparticle zeta potential was found to be extremely negative, measuring - 55.5mV for 18nm and - 31.4mV for 29nm. Behavioural abnormalities, including respiratory distress, reduced responsiveness and erratic swimming, were observed in exposed groups compared to controls, with severity increasing with higher nanoparticle concentrations. Genotoxicity assessment revealed significantly higher DNA damage in kidney cells compared to gill cells. Histological examination of gill tissues showed clogging in primary and secondary lamellae, along with distorted anatomy, necrosis and vacuolar atrophy in peripheral tubules of the kidneys. The kidneys exhibited greater nanoparticle accumulation than the gills with prolonged exposure. Moreover, 18nm AgNPs induced more pronounced DNA damage and histological alterations in the kidney and gill tissues compared to 29nm nanoparticles. This study elucidates the critical role of monitoring AgNPs in aquatic systems, providing essential data on their behaviour and environmental impacts. The findings highlight the need for improved detection techniques and effective management of AgNP contamination. Future research should focus on developing more sensitive analytical methods, understanding long-term ecological effects and exploring innovative remediation strategies.

Evaluation of Genotoxicity by Comet Assay in Tissues of Clarias gariepinus Exposed to Cassava Effluent

Untreated wastewater discharge into waterbodies poses a number of risks, including the potential for aquatic organisms' DNA to be damaged. Using the comet assay, this study assessed the genotoxic impact of cassava wastewater on the gonads, liver, and gills of post-juvenile Clarias gariepinus. The post-juvenile C. gariepinus fish were acquired from a fish farm in Edo State, Nigeria, and were subjected to different concentrations of cassava wastewater (0.2%, 0.3%, 0.5%, and 0.7%) for a duration of 96 hours. In contrast to the typical actions seen in the control groups, the fish exposed to the effluent exhibited restless, erratic movements, and gasping for air. The first set of repetitions showed that catfish mortality increased with concentration; after 96 hours, exposure to the lowest concentration (0.2%) and the highest concentration (0.7%) of cassava wastewater caused 10% and 40% of the fish to die, respectively. The second set of repetitions showed that catfish mortality increased with increasing concentration; after 96 hours, exposure to the lowest concentration (0.2%) and the highest concentration (0.7%) of cassava wastewater resulted in 20% and 50% catfish mortality, respectively. Following the exposure time, the fish were brought to the lab where their gonads, liver, and gills were removed in order to use the comet assay for genotoxic assessment. The genotoxins present in the wastewater can be the reason for the DNA impairment found in the genotoxic assessment result at different effluent concentrations. The investigation also showed that DNA damage increased with increasing concentration, indicating a dose-dependent genotoxic effect of cassava wastewater on post-juveniles of C. gariepinus. This research demonstrates the critical importance of treating wastewater before to discharge.

A comparative account of phototoxicity of anthracene and pyrene in the tadpoles of the anuran amphibian Fejervarya limnocharis using multiple toxicological end points.

Anthracene (Anth) and pyrene (Pyr), two of the priority polycyclic aromatic hydrocarbons (PAHs), being lipophilic in nature, not only accumulate in animals, but also settle in the sediment of water bodies leading to continuous exposure for animals. Anth and Pyr when exposed to sunlight can be photoactivated and have harmful effects on aquatic organisms. A comparative analysis was carried out to assess the acute, sub-chronic, genetic and biochemical toxicity of Anth and Pyr in F. limnocharis tadpoles following short exposures to sunlight on a daily basis. In the bioaccumulation studies, it was found that both Anth and Pyr accumulated in the tadpole tissues in a concentration and time dependent manner. The LC50 values for Anth (under 15 min of daily sunlight exposure) were found to be 2.87, 2.59, 2.28, 1.80 mg/L at 24, 48, 72 and 96 h of the exposures. The corresponding LC50 values for Pyr were 1.03, 0.80, 0.62, 0.42 mg/L. Sublethal exposure of Anth and Pyr affected the survivality, time to metamorphosis as well as morphometric parameters under sunlight exposure. In the genotoxicity assessment studies, particularly the micronucleus test and comet assay, it was found that Pyr led to a higher incidence of micronucleus formation and DNA damage in comparison to Anth. The exposure to PAHs resulted in significant changes in the activity of antioxidant-mediated protective response, specifically the SOD activity, which varied between the groups treated with Anth and Pyr. On the other hand, Pyr treated group showed a higher level of GSH as compared to Anth treated groups. Moreover, the elevation in MDA level in the Anth and Pyr treated groups suggests an increase in lipid peroxidation. Future research should focus on understanding the ecotoxicological risk faced by anuran amphibia due to PAHs that frequently occur in aquatic environments and developing strategies to mitigate these risks.

An evaluation of the genotoxicity and 90-day repeated dose oral toxicity in rats of Porphyridium purpureum.

Interest in microalgae products for use in food is increasing, as demands for sustainable and cost-effective food choices grow due to the escalating global population and increase in climate-related struggles with agriculture. Toxicological assessments of some species of microalgae have been conducted, but there were little data available for the oral consumption of the red microalgae Porphyridium purpureum and no data on genotoxicity. This article articulates a genotoxicity assessment and a 90-day repeated dose oral toxicity study in rats performed according to OECD guidelines. Under the experimental conditions applied, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used in the bacterial reverse mutation test. Similarly, the test item did not induce structural chromosomal aberrations in V79 hamster lung cells. The test item also did not cause chromosomal damage in bone marrow of mice in the mammalian micronucleus test. The no observed adverse effect level (NOAEL) of the 90-day repeated dose oral toxicity study in rats was determined to be the highest dose tested, 3000 mg/kg bw/day. These data add to the body of evidence regarding the safety of P.purpureum for human consumption.

Exploring the Biosafety Potential of Haberlea rhodopensis Friv. In Vitro Culture Total Ethanol Extract: A Comprehensive Assessment of Genotoxicity, Mitotoxicity, and Cytotoxicity for Therapeutic Applications.

The escalating elderly population worldwide has prompted a surge of interest in longevity medicine. Its goal is to interfere with the speed of ageing by slowing it down or even reversing its accompanying effects. As a field, it is rapidly growing and spreading into different branches. One of these is the use of nutraceuticals as anti-ageing drugs. This field is gaining massive popularity nowadays, as people are shifting towards a more natural approach to life and seeking to use natural products as a source of medicine. The present article focuses on the cellular effect of Haberlea rhodopensis Friv. in vitro culture total ethanol extract (HRT), produced by a sustainable biotechnological approach. The extract showed a similar phytochemical profile to plant leaf extract and was rich in primary bioactive ingredients-caffeoyl phenylethanoid glycosides, myconoside, and paucifloside. This study examined the biosafety potential, cytotoxicity, genotoxicity, and mitochondrial activity of the extract using in vitro cultures. The results showed high cell survival rates and minimal cytotoxic effects on Lep3 cells, with no induction of reactive oxygen species nor genotoxicity. Additionally, the extract positively influenced mitochondrial activity, indicating potential benefits for cellular health. The results are promising and show the beneficial effect of HRT without the observation of any adverse effects, which sets the foundation for its further testing and potential therapeutic applications.

Hitting two targets with one shot on pesticide genotoxicity assessment – Identifying risk while unveiling ex vivo approach as a throughput tool in gill-breathing animals

Pesticides in the environment often compromise the ecosystem, thus requiring reliable approaches to assess their effects. Commonly used approaches, such as in vivo, come with several disadvantages, namely in the light of the 3 R’s policy. Seeking for accurate and ethical approaches, this study intended to validate the ex vivo technique as an alternative, and to assess the genotoxicity of chemically-based pesticides and a biopesticide. The ex vivo approach was applied to gill cells of Procambarus clarkii for 2, 4 and 8 h. Cell viability and DNA integrity were evaluated to determine the applicability of this approach. Crayfish gill cells only showed to be suitable for exposures of 2 h. Accordingly, genotoxicity was evaluated in gill cells exposed, for 2 h, to environmentally relevant concentrations of the chemically-based pesticides dimethoate (20 µg L−1), imazalil (160 µg L−1) and penoxsulam (23 µg L−1), as well as to the bioinsecticide Turex® (25, 50, 100, 200 and 400 µg L−1). Every chemically-based pesticide demonstrated to be genotoxic, despite not inducing oxidative DNA damage. On the other hand, Turex® showed no genotoxic effects. Overall, the ex vivo approach demonstrated to be possible and practical to implement, improving the number of outcomes with a lower number of organisms. The findings from the screening test suggest that biological pesticides may pose a lower risk to non-target organisms compared to chemically-based pesticides.

Assessment of acute toxicity, genotoxicity, and anti-inflammatory activity of SteLL, a lectin from Schinus terebinthifolia Raddi. Leaves, in mice

Ethnopharmacology relevanceSchinus terebinthifolia Raddi (Anacardiaceae), known as Brazilian pepper tree, stands out as a medicinal plant widely used in traditional medicine. The leaves are popularly used as anti-inflammatory agent and to relieve inflammatory conditions such as bronchitis, ulcers, and wounds, for example. Aim of the studyThe present study evaluated the acute toxicity, genotoxicity, and anti-inflammatory activity of S. terebinthifolia leaf lectin (SteLL) in mice (Mus musculus). Materials and methodsIn the acute toxicity assay, the animals were treated intraperitoneally (i.p.) or orally (per os) with a single dose of 100 mg/kg. Genotoxicity was assessed by the comet and micronucleus assays. Carrageenan-induced peritonitis and paw edema models were used to evaluate the anti-inflammatory effects of SteLL (1, 5 and 10 mg/kg, i.p.). ResultsNo animal died and no signs of intoxication or histopathological damage were observed in the acute toxicity assay. Genotoxic effect was not detected. In peritonitis assay, SteLL reduced in 56–69% leukocyte migration to the peritoneal cavity; neutrophil count decreased by 25–32%, while mononuclear cell count increased by 67–74%. SteLL promoted a notable reduction of paw edema after 4 h (61.1–63.4%). Morphometric analysis showed that SteLL also decreased the thickness of epidermal edema (30.2–40.7%). Furthermore, SteLL decreased MPO activity, plasma leakage, NO release, and modulated cytokines in both peritoneal fluid and paw homogenate. ConclusionSteLL did not induce acute toxicity or genotoxicity in mice and stands out as a promising candidate in the development of new phytopharmaceuticals with anti-inflammatory action.

Integrated Ozonation and Photocatalysis to Remove Pollutants for Reuse of Rainwater

Rainwater is susceptible to pollutants such as sulphur dioxide, nitrogen oxides, heavy metals, and particles, posing challenges to water quality protection and soil degradation, impacting ecosystems and agriculture. The study focuses on the effectiveness of combined ozonation and photocatalysis in improving physicochemical parameters and reducing toxic substances. Integrated analyses, including ecotoxicological assessments, evaluate the impact of treatment on actual rainwater samples. The results indicate significant reductions in color, heavy metals, and organic pollutants after treatment. Microbiological analyses reveal the inactivation of E. coli, which is crucial for safe water reuse. Ecotoxicity studies show no toxicity to crustaceans, but slight toxicity to algae and bioluminescence bacteria in post-treatment samples. Genotoxicity assessments indicate that there is no detectable DNA damage. Overall, the study highlights the complex nature of rainwater pollution and the efficacy of photocatalytic ozonation in reducing contaminants, underscoring the need for more research to ensure sustainable water resource management.

Genotoxicity and cytotoxicity assessment of 'forever chemicals' in zebrafish (Danio rerio)

Per- and polyfluoroalkyl substances (PFAS) comprise many chemicals with strong carbon-carbon and carbon-fluorine bonds and have extensive industrial applications in manufacturing several consumer products. The solid covalent bonding makes them more persistent in the environment and stays away from all types of degradation, naming them 'forever chemicals.' Zebrafish (Danio rerio) was used to evaluate the genotoxic and cytotoxic effects of legacy PFAS, Perfluorooctane sulfonate (PFOS), and its alternatives, such as Perfluoro-2-methyl-3-oxahexanoic acid ammonium (GenX) and 7H-Perfluoro-3,6-dioxa-4-methyl-octane-1-sulfonic acid (Nafion by-product 2 [NBP2]) upon single and combined exposure at an environmental concentration of 10 µg/L for 48-h. Erythrocyte micronucleus cytome assay (EMNCA) revealed an increased frequency of micronuclei (MN) in fish erythrocytes with a significant increase in NBP2-treated fish. The order of genotoxicity noticed was NBP2 > PFOS > Mixture > GenX in D. rerio. Fish exposed to PFOS and its alternatives in single and combined experiments did not cause any significant difference in nuclear abnormalities. However, PFOS and combined exposure positively inhibit cytokinesis, resulting in an 8.16 and 7.44-fold-change increase of binucleated cells. Besides, statistically, increased levels of reactive oxygen species (ROS) and malondialdehyde (MDA) content indicate oxidative stress in D. rerio. In addition, 'forever chemicals' resulted in cytotoxicity, as evident through changes in nucleus width to the erythrocyte length in NBP2 and mixture exposure groups. The findings revealed that PFAS alternative NBP2 is more toxic than PFOS in inducing DNA damage and cytotoxicity. In addition, all three tested 'forever chemicals' induced ROS and lipid peroxidation after individual and combined exposure. The present work is the first to concern the genotoxicity and cytotoxicity of 'forever chemicals' in the aquatic vertebrate D. rerio.

Multi-Endpoint Toxicological Assessment of Chrysin Loaded Oil-in-Water Emulsion System in Different Biological Models.

Chrysin is hypothesized to possess the ability to prevent different illnesses, such as diabetes, cancer, and neurodegenerative disorders. Nonetheless, chrysin has a low solubility under physiological conditions, resulting in limited bioavailability. In a previous study, we utilized an oil-in-water emulsion system (chrysin-ES or chrysin-NE) to encapsulate chrysin, thereby increasing its bioaccessibility and preserving its antioxidant and anti-Alzheimer's properties. To promote the chrysin-ES as a supplementary and functional food, it was obligatory to carry out a safety assessment. Cytotoxicity testing showed that chrysin-ES was harmless, with no killing effect on 3T3-L1 (adipocytes), RAW 264.7 (macrophages), HEK293 (kidney cells), and LX-2 (hepatic stellate cells). The acute toxicity evaluation demonstrated that the 50% lethal dose (LD50) for chrysin-ES was greater than 2000 mg/kg BW. Genotoxicity assessments found that chrysin-ES did not induce DNA mutations in vitro or in vivo. Furthermore, chrysin and chrysin-ES exhibited anti-mutagenic properties against PhIP-induced and IQ-induced mutagenesis in the Ames test, while they inhibited urethane-, ethyl methanesulfonate-, mitomycin C-, and N-nitrosomethylurea-mediated mutations in Drosophila. The present study illustrates the safety and anti-genotoxicity properties of chrysin-ES, allowing for the further development of chrysin-based food supplements and nutraceuticals.

Challenges of the Application of In Vitro Digestion for Nanomaterials Safety Assessment.

Considering the increase in the production and use of nanomaterials (NM) in food/feed and food contact materials, novel strategies for efficient and sustainable hazard characterization, especially in the early stages of NM development, have been proposed. Some of these strategies encompass the utilization of in vitro simulated digestion prior to cytotoxic and genotoxic assessment. This entails exposing NM to fluids that replicate the three successive phases of digestion: oral, gastric, and intestinal. Subsequently, the resulting digestion products are added to models of intestinal cells to conduct toxicological assays, analyzing multiple endpoints. Nonetheless, exposure of intestinal cells to the digested products may induce cytotoxicity effects, thereby posing a challenge to this strategy. The aim of this work was to describe the challenges encountered with the in vitro digestion INFOGEST 2.0 protocol when using the digestion product in toxicological studies of NM, and the adjustments implemented to enable its use in subsequent in vitro biological assays with intestinal cell models. The adaptation of the digestion fluids, in particular the reduction of the final bile concentration, resulted in a reduced toxic impact of digestion products.

Assessment of genotoxicity of air pollution in urban areas using an integrated model of passive biomonitoring

Atmospheric pollution is a major public health issue and has become increasingly critical for human health. Urban atmospheric pollution is typically assessed through physicochemical indicators aligned with environmental legislation parameters, providing data on air quality levels. While the effects of pollution on sensitive organisms serve as a warning for public health decision-makers, there remains a need to explore the interpretation of environmental data on pollutants. The use of species adapted to urban environments as sentinels enables continuous and integrated monitoring of environmental pollution implications on biological systems. In this study, we investigated the use of the plant species Tradescantia pallida as a biomonitor to evaluate the genotoxic effects of atmospheric pollution under diverse vehicular traffic conditions. T. pallida was strategically planted at the leading urban intersections in Uberlândia, Brazil. During COVID-19 pandemic lockdowns, we compared indicators such as physical, biological, and traffic data at different intersections in residential and commercial zones. The reduction in vehicular traffic highlighted the sensitivity of plant species to changes in air and soil pollutants. T. pallida showed bioaccumulation of heavy metals Cd and Cr in monitored areas with higher traffic levels. Additionally, we established a multiple linear regression model to estimate genotoxicity using the micronucleus test, with chromium concentration in the soil (X1) and particulate matter (PM) in the atmosphere (X2) identified as the primary independent variables. Our findings provide a comprehensive portrait of the impact of vehicular traffic changes on PM and offer valuable insights for refining parameters and models of Environmental Health Surveillance.

Pobody’s Nerfect: (Q)SAR works well for predicting bacterial mutagenicity of pesticides and their metabolites, but predictions for clastogenicity in vitro have room for improvement

Genotoxicity assessment is a key component of regulatory decision-making in pesticide authorization and biocide approval. Conventionally, these genotoxicity requirements are addressed with OECD test guideline-compliant in vitro tests. In recent years, in silico approaches, such as (Q)SAR, have matured sufficiently so that they may be suitable to support, complement or even replace in vitro testing as a first tier of genotoxicity assessment. Among the different endpoints for genotoxicity, a high reliability is expected for in silico predictions of the endpoint bacterial mutagenicity. For other endpoints predictive performance is either unclarified or seems to be comparably lower. Herein, we describe the evaluation of several commercial and freely available (Q)SAR models and complementary combinations thereof with respect to the endpoints bacterial mutagenicity and chromosome damage in vitro. We used curated in-house test sets derived from OECD test guideline-compliant studies, gathered from submissions for the regulatory approval of biocides and plant protection products. The data set comprises active substances, metabolites and impurities. In line with previous publications we show that (Q)SAR models for bacterial mutagenicity generally performed well for compounds of the pesticide domain. Model combinations significantly increased the respective sensitivity. Models for chromosome damage still need to improve prior to their stand-alone use in regulatory decision-making, either strongly leaning towards sensitivity, at the expense of specificity or vice versa. Similar to the endpoint bacterial mutagenicity, combinations of models for chromosome damage increase sensitivity when compared to the individual models alone.

Oral toxicity and genotoxicity assessment of standardized Echinacea purpurea (L.) extract and the pharmacokinetic profile of its active ingredient chicoric acid

Oral toxicity and genotoxicity assessment of standardized Echinacea purpurea (L.) extract and the pharmacokinetic profile of its active ingredient chicoric acid

Assessment of Ingested Micro- and Nanoplastic (MNP)-Mediated Genotoxicity in an In Vitro Model of the Small Intestinal Epithelium (SIE).

Micro- and nanoplastics (MNPs) have become ubiquitous contaminants of water and foods, resulting in high levels of human ingestion exposure. MNPs have been found in human blood and multiple tissues, suggesting that they are readily absorbed by the gastrointestinal tract (GIT) and widely distributed. Growing toxicological evidence suggests that ingested MNPs may pose a serious health threat. The potential genotoxicity of MNPs, however, remains largely unknown. In this study, genotoxicity of primary and environmentally relevant secondary MNPs was assessed in a triculture small intestinal epithelium (SIE) model using the CometChip assay. Aqueous suspensions of 25 and 1000 nm carboxylated polystyrene spheres (PS25C and PS1KC), and incinerated polyethylene (PEI PM0.1) were subjected to simulated GIT digestion to create physiologically relevant exposures (digestas), which were applied to the SIE model at final MNP concentrations of 1, 5, and 20 μg/mL for 24 or 48 h. PS25C and PS1KC induced DNA damage in a time- and concentration-dependent manner. To our knowledge, this is one of the first assessment of MNP genotoxicity in an integrated in vitro ingestion platform including simulated GIT digestion and a triculture SIE model. These findings suggest that ingestion of high concentrations of carboxylated PS MNPs could have serious genotoxic consequences in the SIE.

Scientific Opinion on additional scientific data related to the safety of preparations of Rheum palmatum L., Rheum officinale Baill. and their hybrids, Rhamnus purshianaDC., Rhamnus frangula L. and Cassia senna L., submitted pursuant to Article 8(4) of Regulation (EC) No 1925/2006.

The Panel on Nutrition, Novel Foods and Food Allergens (NDA) was asked to deliver a scientific opinion on the safety of plant preparations from the root or rhizome of Rheum palmatum L., Rheum officinale Baill. and their hybrids, from the bark of Rhamnus frangula L. and Rhamnus purshiana DC. and from the leaf or fruit of Cassia senna L., which have been placed under Union scrutiny in Part C of Annex III in accordance with Article 8(4) of Regulation (EC) No 1925/2006. The NDA Panel reviewed the additional scientific data submitted during the period of scrutiny and the public consultation by interested parties. The pertinent scientific data were invitro and invivo genotoxicity studies on the plant preparations under consideration. All the results of the genotoxicity studies on plant preparations were negative. However, the plant preparations that were tested in the submitted studies were not sufficiently characterised with respect to the content of total and individual hydroxyanthracene derivatives (HADs) and components other than HADs. The studies confirmed the presence of ■■■■■, known to be genotoxic invivo, and ■■■■■, shown to be genotoxic invitro. In line with the EFSA Scientific Committee statement on genotoxicity assessment of chemical mixtures, considering the presence of an invivo genotoxic compound, the plant preparations used in these studies have to be considered of concern for genotoxicity. Thus, the safety of preparations containing HADs from the root or rhizome of Rheum palmatum L., Rheum officinale Baill. and their hybrids, from the leaf or fruit of Cassia senna L. and from the bark of Rhamnus frangula L. and Rhamnus purshiana DC. cannot be established based on the submitted studies.

Forced Degradation Products of Olmesartan Medoxomil and their In Vitro Genotoxicity Assessment by Comet Assay using HEK Cells

AbstractThis study designed to characterize major forced degradation products of Olmesartan Medoxomil (OM) and to perform genotoxicity profiling using acid, base, and hydrogen peroxide‐induced stress degradation studies. The isolation of degradation products was conducted on flash chromatography using FlashPure ECOFLEX C18 4 g column with gradient elution and diode array detection‐based fraction collection, and characterized by mass spectrometry. Three impurities (impurity‐1, 2, and 3) were identified with respective m/z values of 447.213, 461.229, and 463.208. The genotoxic potential of impurities was assessed by in silico prediction, and in vitro comet assay using HEK cells. The extent of DNA damage was analyzed using Comet Assay IV imaging software. For impurity‐1, head and tail lengths and their respective total intensities were significantly (p<0.001) larger than those of dimethyl sulfoxide (solvent) control and comparable to positive control. Other parameters such as mean grey level and total area of comet were statistically significant as compared with the solvent and positive controls. Overall, impurity‐1 was a possible genotoxic impurity and a known major degradation product in OM tablet dosage form. In silico prediction also revealed impurity‐1 as toxicity class 3, whilst impurities 2 and 3 were of toxicity class 4.