Oral Toxicity In Rats Research Articles

A 13-week repeated oral dose toxicity evaluation and a 4-week recovery evaluation of rosemary concentrate containing 50% ursolic acid in male and female rats.

This study was performed to evaluate the safety of rosemary concentrate containing 50% ursolic acid (RCUA50), the ethanolic extract of rosemary. RCUA50 was administered orally for 13 weeks at 1000, 2000, and 4000 mg/kg/day, and then the rats were maintained for 4 weeks without RCUA50 administration for recovery evaluation. We observed clinical signs, body weights, food consumption, functional observations, ophthalmological examination, urinalysis, estrus cycle, hematology, clinical chemistry, sperm analysis, organ weights, gross examination, and histopathological examinations. During the dosing and recovery period, there were no test substance-related deaths, clinical signs, changes of body weights, and food consumption in all treated groups. In the main group, there were no test substance-related effects in functional observations and ophthalmological examination. In the main and recovery groups, there were no test substance-related effects in hematology, clinical chemistry, sperm examination, organ weights, necropsy and histopathological examination. In conclusion, the repeated oral administration of RCUA50 for 13 weeks resulted in no test substance-related adverse effect at all dose levels. Therefore, the NOAEL was considered to be greater than 4,000 mg/kg/day in both sexes under the conditions of this study.

GC-MS Analysis, Oral Toxicity Evaluation, and Molecular Docking Studies of Ocimum tenuiflorum L. Essential Oil: Exploring its Anti-Diabetic Potential through Peroxisome Proliferator-Activated Receptor-Delta (PPARδ) Activation

Ocimum tenuiflorum (O. tenuiflorum), commonly known as Tulsi, is an aromatic herb with significant medicinal properties, particularly in the management of diabetes. This study aimed to extract essential oil from the leaves of O. tenuiflorum, perform a comprehensive GC-MS analysis, evaluate acute oral toxicity, and assess its anti-diabetic potential through computational molecular docking with the peroxisome proliferator-activated receptor-delta (PPARδ) protein. The essential oil was extracted using hydro-distillation, and GC-MS identified 28 compounds, with (E)-caryophyllene, β-elemene, and trans-isoeugenol being the most abundant. Molecular docking against PPARδ (PDB ID: 5U3R) highlighted rimuene, himachalane, valencene, and nootkatene as top candidates, exhibiting strong binding affinities comparable to the reference drug seladelpar. Predominantly hydrophobic interactions were observed due to the volatile, non-polar nature of the compounds in the oil. Acute oral toxicity tests showed an LD50 >2000 mg/kg body weight which confirmed the safety of the essential oil, with low toxicity profile. This study identifies rimuene and himachalane as promising PPARδ modulators, suggesting further in vitro, in vivo, and computational evaluations to validate their therapeutic potential in diabetes management.

Safety Evaluation for Acute and Chronic Oral Toxicity of Maha Pigut Triphala Contains Three Medicinal Fruits in Sprague-Dawley Rats.

Maha Pigut Triphala is the herbal mixture of three fruits consisting of T. bellirica, T. chebula, and E. officinalis also known as P. emblica. Humans regularly eat the fresh fruits of these plants on a daily basis. Maha Pigut Triphala is one of the widely known herbal medicinal formulas used in traditional Thai medicine. Besides studying pharmacological properties, attention should also be paid to the safety and toxicity studies of herbal medicines. The objective of the present study was to evaluate the acute and chronic oral toxicity of Maha Pigut Triphala (2:1:3) in Sprague-Dawley rats. A single dose of Maha Pigut Triphala at a concentration of 5000 mg/kg body weight was administered orally to female rats in the acute oral toxicity study. In the chronic oral toxicity study, male and female rats were treated with various concentrations of Maha Pigut Triphala (600, 1200, and 2400 mg/kg body weight) once daily for 270 consecutive days. The presence of abnormalities in the symptoms and behavior of the rats were observed and recorded throughout the experiment. Additionally, body weight, organ weight, and mortality were recorded. At the end of the study, blood samples were collected for hematological and blood chemistry analysis, while the internal organs were evaluated for gross pathological and histopathological changes. The acute oral toxicity study revealed no mortality and abnormal symptoms or behavior in Maha Pigut Triphala-treated rats. Moreover, gross pathological and histopathological findings did not reveal any abnormalities in the internal organs. In the chronic oral toxicity evaluation, although there were negligible changes in body weight, organ weight, and hematological and blood chemistry parameters in rats treated with Maha Pigut Triphala for 270 days, no behavioral or gross pathological and histopathological abnormalities were observed. Overall, the results of this study demonstrate that Maha Pigut Triphala (2:1:3) neither causes acute nor chronic oral toxicity in rats, proposing the safety of this herbal formula in animals prior to human trials and use.

Cell Painting and Chemical Structure Read-Across Can Complement Each Other for Rat Acute Oral Toxicity Prediction in Chemical Early Derisking.

Early derisking decisions in the development of new chemical compounds enable the identification of novel chemical candidates with improved safety profiles. In vivo studies are traditionally conducted in the early assessment of acute oral toxicity of crop protection products to avoid compounds, which are considered "very acutely toxic", with an in vivo lethal dose of 50% (LD50) ≤ 60 mg/kg body weight. Those studies are lengthy and costly and raise ethical concerns, catalyzing the use of nonanimal alternatives. The objective of our analysis was to assess the predictive efficacy of read-across approaches for acute oral toxicity in rats, comparing the use of chemical structure information, in vitro biological data derived from the Cell Painting profiling assay on U2OS cells, or the combination of both. Our findings indicate that the classification of compounds as very acute oral toxic (LD50 ≤ 60 mg/kg) or not is possible using a read-across approach, with chemical structure information, morphological profiles, or a combination of both. When classifying compounds structurally similar to those in the training set, the chemical structure was more predictive (balanced accuracy of 0.82). Conversely, when the compounds to be classified were structurally different from those in the training set, the morphological profiles were more predictive (balanced accuracy of 0.72). Combining the two models allowed for the classification of compounds structurally similar to those in the training set to slightly improve the predictions (balanced accuracy of 0.85).

A toxicological assessment of Ganoderma lucidum and Cordyceps militaris mushroom powders.

Gonoderma lucidum (G. lucidum) and Cordyceps militaris (C. militaris) are among the many mushrooms known for their long history of use in traditional medicine. Wildcrafted sources of mushrooms including G. lucidum and C. militaris can be limited from a scarcity and quality perspective, but solid fermentation processes in cultivation settings can provide an efficient way to deliver whole mushroom preparations of a consistent composition. Despite the historical use of these mushrooms, few published reports have explored their potential subchronic oral toxicity or genotoxicity, either from specific components or whole mushroom preparations. The purpose of this study was to assess the potential for acute toxicity, subchronic toxicity, and genotoxicity of powders produced from G. lucidum mycelial biomass and fruiting body ("Organic Reishi M2-102-02 powder") cultured on oats, and C. militaris mycelial biomass, stroma, and fruiting body ("Organic Cordyceps M2-116-04 powder") cultured on oats. Results of the testing demonstrate that both Organic Reishi M2-102-02 powder and Organic Cordyceps M2-116-04 powder were not acutely toxic, did not exhibit subchronic oral toxicity in rats at doses up to the highest dose tested of 2,000mg/kgbw/day, and did not have genotoxic potential based on in vitro and in vivo genotoxicity assays.

Evaluating the performance of multi-omics integration: a thyroid toxicity case study

Multi-omics data integration has been repeatedly discussed as the way forward to more comprehensively cover the molecular responses of cells or organisms to chemical exposure in systems toxicology and regulatory risk assessment. In Canzler et al. (Arch Toxicol 94(2):371–388. https://doi.org/10.1007/s00204-020-02656-y), we reviewed the state of the art in applying multi-omics approaches in toxicological research and chemical risk assessment. We developed best practices for the experimental design of multi-omics studies, omics data acquisition, and subsequent omics data integration. We found that multi-omics data sets for toxicological research questions were generally rare, with no data sets comprising more than two omics layers adhering to these best practices. Due to these limitations, we could not fully assess the benefits of different data integration approaches or quantitatively evaluate the contribution of various omics layers for toxicological research questions. Here, we report on a multi-omics study on thyroid toxicity that we conducted in compliance with these best practices. We induced direct and indirect thyroid toxicity through Propylthiouracil (PTU) and Phenytoin, respectively, in a 28-day plus 14-day recovery oral rat toxicity study. We collected clinical and histopathological data and six omics layers, including the long and short transcriptome, proteome, phosphoproteome, and metabolome from plasma, thyroid, and liver. We demonstrate that the multi-omics approach is superior to single-omics in detecting responses at the regulatory pathway level. We also show how combining omics data with clinical and histopathological parameters facilitates the interpretation of the data. Furthermore, we illustrate how multi-omics integration can hint at the involvement of non-coding RNAs in post-transcriptional regulation. Also, we show that multi-omics facilitates grouping, and we assess how much information individual and combinations of omics layers contribute to this approach.

First insight into the formation of transformation products of a biopesticide guvermectin in rat and its health risk

Guvermectin is a new chemical isolated from the microbial metabolites and is registered as a novel plant growth regulator. However, the biotransformation behavior and toxicity of guvermectin to mammals remain unclear and have unknown implications for consumers or occupationally exposed persons. Therefore, we investigated the biotransformation of guvermectin in vivo and in vitro, its effects on CYP450s activities, and its oral toxicity in rats. The results showed that guvermectin could be rapidly absorbed when administered orally and eliminated rapidly in the serum, with a half-life of 6.3 h. Four phase І metabolism products of guvermectin in the serum were screened and identified using UPLC-QTOF/MS. Two products, adenine and psicofuramine, were confirmed using reference standards. Hydrolysis and oxidation reactions were the main transformation pathways. Oral toxicity tests in rats showed that guvermectin exhibited light toxicity to rats (LC50 > 5000 mg/kg b.w.). However, an in vitro probe drug experiment revealed that guvermectin could induce CYP2D6 activity, and a lower concentration of guvermectin exhibited a stronger effect on CYP2D6 than higher concentration (1.38-fold). Molecular docking studies implied that guvermectin was an antagonist of CYP1A2, CYP2C9, and CYP3A4. These findings provided a better understanding of the environmental and human health risks associated with guvermectin and promote its rational use. However, the potential risk of endocrine disruption can not be ignored due to the presence of nucleoside-like metabolites.

Bifidobacterium longum subsp. infantis YLGB-1496-Toxicological evaluation.

Bifidobacterium infantis YLGB-1496, originally isolated from breast milk from a Taiwanese mother, is under study for use as a probiotic. As part of safety assessment, an Ames, in vivo mouse micronucleus, and in vivo mouse spermatocyte chromosome aberration assay were conducted along with a 13-week oral rat toxicity study. B.infantis YLGB-1496 had no activity in any of the genotoxicity assays. Administration of the bacteria to Sprague-Dawley rats at doses ranging from 0 to 1.5 g/kg bw/day had no treatment-related effects on any of the endpoints measured. There appear to be no concerns for translocation or pathogenicity of B.infantis YLGB-1496 based on extensive experience with the species in general. The results of the current investigations support potential use of B.infantis YLGB-1496 as a probiotic in infant formula.

Acute and subacute oral toxicity evaluation of Ayurvedic formulation Tapyadi loha in rats

Ayurveda is one of the oldest systems of traditional medicine that provides treatments for a wide range of acute and chronic health problems. It is a common myth amongst people that Ayurvedic drugs have no side effects, whereas the fact is that these drugs can cause adverse effects. Despite their wide use, the safety data of many Ayurvedic formulations are still unavailable. Tapyadi loha is an Ayurvedic formulation traditionally claimed for iron deficiency anemia in pregnant and non-pregnant patients. However, no scientific study has been conducted to evaluate its oral toxicity. Hence, the present study evaluated the acute and subacute oral toxicity of the Tapyadi loha according to the OECD test guidelines 425 and 407, respectively. Tapyadi loha did not cause mortality nor any signs of toxicity when given once orally at a dose of 2000 mg/kg. Subacute toxicity study showed no mortality as well as no behavioral, hematological, biochemical and histopathological abnormalities in rats treated with Tapyadi loha formulation at 250, 500 and 1000 mg/kg for 28 days. It is concluded that the Tapyadi loha is safe at a single dose of 2000 mg/kg and 28 days repeated dose of 1000 mg/kg by oral route in rats.

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.

Evaluation of Acute and Repeated Dose Oral Toxicity of Phoenix dactylifera L. Pollen Methanolic Extract in Rats

Pollen of the date palm (Phoenix dactylifera L.) has been used for thousands of years as a traditional herbal medicine for improving fertility. However, safety of plant-derived materials is an important concern to human health, and herbal remedies should be taken with adequate knowledge about their toxicity, adverse effects, purity, appropriate dosage and dose limits. The current work was undertaken to evaluate the safety of date palm pollen (DPP) and to classify it according to its oral toxicity in rats. The experiments were done according to the reported guidelines of the Organization for Economic Co-operation and Development (OECD) to assess the acute and repeated dose oral toxicity in rats. The obtained results showed neither death nor sign of toxicity or abnormality in any of the animals throughout the experimental period regarding acute and repeated dose oral toxicity. Also the results of the hematological and clinical biochemistry determinations showed no significant differences between the control and DPP-treated rat groups. It could be concluded that, the tested plant material is a relatively safe substance and its approximate LD50 defined by 2000mg/kg < LD50 < 5000mg/kg according to the OECD tests guidelines.

Spectral characterization, analgesic, and anti-inflammatory effects of ethanolic extract of Calotropis procera leaf and dry latex from Jazan, Kingdom of Saudi Arabia

Traditional healers have used the shrub Calotropis procera (CP) for many years for various therapies. The present study investigated the bioactive constituents of ethanolic extract of CP leaf and dried latex using gas chromatography-mass spectrometry and Fourier transforms infrared spectroscopy. The identification and characterization of the compounds were confirmed by examining the constituents' mass spectrum fragmentations and FT-IR spectra and comparing the results with those in the literature. The tail-flick method was used to investigate the analgesic properties of the extract and its anti-inflammatory activities using a rat model of formalin-induced oedema. Acute oral toxicity in rats was studied per OECD recommendations. Twenty male rats were divided into four groups and received an ethanolic extract of the leaves and dried milky sap of CP (200 mg/Kg) in groups 1, 2, and 3. Group 4 rats were administered aspirin 50 mg/kg as a positive control. The CP dried latex extract has the highest content of lupeol and its acetate derivative compared to its leaf extract. The CP dried latex extract inhibited inflammation more significantly than the ethanolic leaf extract and the drug indomethacin at a higher dosage (200 mg/kg). The ethanolic extracts showed analgesia comparable to aspirin. It suggests that fatty acids and their esters, particularly ethyl linoleate (8.96%), ethyl palmitate (7.99%), ethyl linoleate (6.98%), and palmitic acid (5.18%), may be valuable biomarkers for characterizing leaf and latex samples and describing the medicinal potential of CP.

Biochemical parameters, oxidative stress biomarkers, and anatomopathological changes in Wistar rats treated with 3′-demethoxy-6-O-demethylisoguaiacin and norisoguaiacin

Artemisia cina (Ac) is a plant with anthelmintic compounds such as 3′-demethoxy-6-O-demethylisoguaiacin (D) and norisoguaiacin (N). Three major objectives were proposed: (1) To evaluate biochemical parameters in blood (2) to determine the tissue oxidative stress by biomarkers as TBARS and glutathione peroxidase activity, and (3) to evaluate anatomopathological changes in organs such as the brain, liver, kidney, and lung after oral administration of n-hexane extract of Ac and D and N. D and N were administrated following the OECD guides for acute oral toxicity evaluation (Guide 420). Fifty Wistar rats were distributed into ten groups as follows: Group 1 (G1): 4 mg/Kg; G2: 40 mg/Kg; G3: 240 mg/Kg; G4: 1600 mg/Kg of n-hexane extract of Ac. G5: 2 mg/Kg; G6: 20 mg/Kg; G7: 120 mg/Kg; G8: 800 mg/Kg of D and N, G9: water and G10: polyvinylpyrrolidone at 2000 mg/Kg. At 14 days, the rats were euthanized, and the blood, liver, brain, kidney, and lung were taken for biochemical analysis, anatomopathological changes, and TBARS and GSH evaluation. Glucose, cholesterol, and phosphorus were altered. Histopathological analysis showed multifocal neuronal degeneration in the brain (G2). The kidney and lungs had changes in G7. The GSH and TBARS increased in G6 and G7. The TBARS activity was higher in G1 and G2. In conclusion, extract and D and N of Ac did not have damage at therapeutic doses. D, N, and n-hexane extract of A. cina do not cause histopathological damage at pharmaceutical doses. Still, the brain, kidney, and liver are related to biochemical parameters at higher doses. However, compounds are proposed as antioxidant agents.

Safety study of carboxymethylated Basella alba mucilage: a subchronic oral toxicity evaluation in Wistar albino rats

Background: The evaluation of toxicity is of paramount importance in the screening of a new compound. Basella alba mucilage possesses a versatile excipient property that can be innovated with its chemical modification to get the functionalized mucilage. A few pharmacological activities of Basella alba mucilage have also been reported earlier, but its toxicity study in rats has yet to be discussed. Aim: The study aims to assess the in vivo toxicity of carboxymethylated Basella alba mucilage in Wistar albino rats for 28 days. Material and Methods: In the current investigation, Carboxymethylated Basella alba mucilage is taken, and its subchronic toxicity study is carried out in forty-eight healthy rats (twenty-four male rats and 24 female rats), divided into four groups containing six rats of each sex. All the biochemical and hematological parameters and histopathological investigation were estimated for all the animal groups. Result: The subchronic toxicity study reveals that the modified mucilage is safe for all doses (20mg/kg body weight, 40mg/kg body weight, 80 mg/kg body weight). The study showed no significant difference in the dose group's behavioral toxicity, nephrotoxicity, and hepatotoxicity compared to the control. All the hematological and biochemical parameters lay in the normal range. The histopathological examination of treatment groups showed no abnormality or lesion in the tissue samples of internal organs. Conclusion: The study confirms the safety of Carboxymethylated Basella alba mucilage for use in pharmaceutical formulations.

Characterization of the antioxidant activity, identified free radical-relieving components by LC/QTOF/MS and acute oral toxicity studies of Tri-Tharn-Thip tea

This study assessed the antioxidant activities and bioactive maker of Tri-Than-Thip tea (Tri-TTa) to provide database support for the use of Tri-Than Thip remedy according to the Thai Pharmaceutical Textbook and aims to develop herbal medicine or food from natural products. Tri-TTa and herbal components were tested for antioxidant scavenging activity (DPPH assay, ABTS assay), chain-breaking/iron ion chelating activity (Metal Chelating Activity; MAC, the ferric-reducing antioxidant power activity; FRAP), superoxide anion radical scavenging activity (NBT assay), oxygen radical antioxidant capacity (ORAC assay), total content of Phenolic (TCP) and flavonoid (TCP) substance. The chemical components were separated and investigated by LC/QTOF/MS for safe dosages due to acute oral toxicity in rats. The Tri-TTa showed the strongest inhibition of antioxidant activity with the DPPH, ABTS, and MCA assay, with IC50 values of 0.087 ± 0.003, 0.025 ± 0.000, and 0.012 ± 0.000 mg/mL, respectively. In addition, Tri-TTa can also inhibit superoxide anion radicals, with IC50 values of 158 ± 0.009 mg/mL, and the oxygen radical was 3.47 ± 0.03 μM of TE/g extract. TCP, TCF, and FRAP values of Tri-TTa were 154.123 ± 1.326, 80.233 ± 14.268 mg equivalent/g of extract, and 1092.457 ± 106.024 mM FeSO4/mg, respectively. Additionally, Tri-TTa contains important substances in the phenolic and flavonoid groups that have good antioxidant abilities and were confirmed to be safe through acute oral toxicity testing in rats, which revealed no toxicity-related effects at high doses. Tri-TTa is a strong candidate for use as a natural product with antioxidant activity, suggesting that plant polyphenol content may promote antioxidant activity. These results support further research on Thai folk remedy infusions as promising antioxidant agents that can be developed for food, tea, or tonic drugs from natural.

Oral acute toxicity study of ethanol extract of Mobe leaves (Artocarpus lacucha Buch-Ham) in Wistar rats

Many medicinal plants are now being chosen because the treatment is safer and cheaper. Artocarpus lacucha Buch-Ham is a plant with many pharmacological activities and is efficacious; its safety has not been studied. Acute oral toxicity evaluation followed Organization for Economic Co-operation and Development guidelines using the fixed dose method. The evaluation began with a preliminary test, then a primary test with three groups: a 2000 mg/Kg BW dose test group, a 5000 mg/Kg BW dose test group, and a control group. The results of visual observations, haematological and clinical examinations, and histological examination of organs (liver, spleen, kidneys, lungs and heart) showed no toxicity in the animals, and they did not die during testing. The findings of this study support the safety of Artocarpus lacucha Buch-Ham leaf ethanol extract, which did not produce harmful results in acute toxicity tests.

Replacing concurrent controls with virtual control groups in rat toxicity studies

Virtual control groups (VCGs) in nonclinical toxicity represent the concept of using appropriate historical control data for replacing concurrent control group animals. Historical control data collected from standardized studies can serve as base for constructing VCGs and legacy study reports can be used as a benchmark to evaluate the VCG performance. Replacing concurrent controls of legacy studies with VCGs should ideally reproduce the results of these studies. Based on three four-week rat oral toxicity legacy studies with varying degrees of toxicity findings we developed a concept to evaluate VCG performance on different levels: the ability of VCGs to (i) reproduce statistically significant deviations from the concurrent control, (ii) reproduce test substance-related effects, and (iii) reproduce the conclusion of the toxicity study in terms of threshold dose, target organs, toxicological biomarkers (clinical pathology) and reversibility. Although VCGs have shown a low to moderate ability to reproduce statistical results, the general study conclusions remained unchanged. Our results provide a first indication that carefully selected historical control data can be used to replace concurrent control without impairing the general study conclusion. Additionally, the developed procedures and workflows lay the foundation for the future validation of virtual controls for a use in regulatory toxicology.

The short-term toxicity and metabolome of dicyclopentadiene

Dicyclopentadiene (DCPD) was investigated in a 14-day oral rat toxicity study based on the OECD 407 guideline in combination with plasma metabolomics. Wistar rats received the compound daily via gavage at dose levels of 0, 50 and 150 mg/kg bw. The high dose induced transient clinical signs of toxicity and, in males only, reduced body weight gain. High dose liver changes were characterized by altered clinical chemistry parameters in both sexes and pathological changes in females. In high dose males an accumulation of alpha-2 u-globulin in the kidney was noted. Comparing the DCPD metabolome with previously established specific metabolome patterns in the MetaMap® Tox data base suggested that the high dose would result in liver enzyme induction leading to increased breakdown of thyroid hormones for males and females. An indication for liver toxicity in males was also noted. Metabolomics also suggested an effect on the functionality of the adrenals in high dose males, which together with published data, is suggestive of a stress related effect in this organ. The results of the present 14-day combined toxicity and metabolome investigations were qualitatively in line with literature data from subchronic oral studies in rats with DCPD. Importantly no other types of organ toxicity, or hormone dysregulation beyond the ones associated with liver enzyme induction and stress were indicated, again in line with results of published 90-day studies. It is therefore suggested that short term “smart” studies, combining classical toxicity with ‘omics technologies, could be a 2 R (refine and reduce) new approach method allowing for the reduction of in vivo toxicity testing.

Cytotoxicity, Antiadipogenic, Low‐Density Lipoprotein Oxidation Inhibitory Activities, and Acute Toxicity Study of Psychotria densinervia Hydroethanolic Leaf and Bark Extracts

Background: Obesity is increasingly taking an important stage as a cause of death worldwide, and interventions with a good cost‐effectiveness ratio are needed. Psychotria densinervia is one of these natural products with health benefits. Objective. The present study evaluated the cytotoxicity, antiadipogenic, low‐density lipoprotein (LDL), oxidation inhibitory activities, and acute toxicity of Psychotria densinervia hydroethanolic leaf and bark extracts.Methods: The cytotoxicity evaluation of the extracts (62.5, 125, 250, and 500 μg/mL) using the MTT assay and the antiadipogenic activity (25, 50, 100, and 200 μg/mL) using oleic acid were carried out in SW‐872 cells. Copper sulfate (CuSO4)‐induced oxidation was used in the evaluation of the effect of extracts (0.25, 0.5, and 1 mg/mL) against LDL oxidation. The oral acute toxicity evaluation of a single dose of 2000 mg/kg of the extracts was performed in Wistar albino rats weighing 127 ± 2 g.Results: The leaf and bark extracts did not show any sign of cytotoxicity at the tested concentrations. The best antiadipogenic activity was observed by the standard orlistat (38.45 ± 1.70 μg/mL), followed by the leaf extract (IC50: 41.47 ± 0.50 μg/mL) and the least the bark extract (IC50: 107.50 ± 0.90 μg/mL). At a concentration of 1 mg/mL, the leaf extract presented an oxidation lag time of 130 min, which was higher and better than that of the bark extract (120 min). Quercetin (standard) presented an oxidation lag time longer than 3 h. The oral acute toxicity evaluation did not show any signs of toxicity indicating that the LD50 was greater than 2000 mg/kg.Conclusion: Based on the results obtained, the P. densinervia hydroethanolic leaf extract possesses a better antioxidant and antiadipogenic activities than the bark extract.

Toxicological Evaluation of a Polyherbal Formulation (18KHT01) and Validation of UPLC-DAD Method for Quality Control.

Background: 18KHT01 is a novel synergistic composition of Quercus acutissima, Camellia sinensis, Geranium thunbergii, and a small portion of Citrus limon. Our previous report demonstrated that the 18KHT01 exhibits potent antiobesity effects, with synergistic antioxidant, antiadipogenic, and antiobesity activities in diet-induced obese mice. This study explores the toxicity profile and quality control parameters of the 18KHT01 formulation. Methods: Broad-spectrum acute and subacute oral toxicity studies were performed using male and female ICR mice. In order to simultaneous analysis of the 18KHT01 formulation, an ultraperformance liquid chromatography coupled with a diode array detector (UPLC-DAD) method was developed and validated using six marker compounds. Results: Acute oral toxicity evaluation of 18KHT01, administered at single high doses of 2, 2.5, 3, and 5 g/kg, identified 2 g/kg as the no-observed adverse effect level (NOAEL). The LD50 (50% lethal dose) and LD100 (100% lethal dose) of 18KHT01 for male ICR mice were 3.99 and 7.77 g/kg, and those for the female mice were 2.94 and 4.70 g/kg, respectively. In addition, a 30-day repeated dose oral subacute toxicity evaluation indicated that 18KHT01 is safe below 500 mg/kg/day for long-term administration in ICR mice of either sex. UPLC-DAD method validation revealed that each calibration curve for the marker compounds showed good linearity, as well as the validation parameters such as precision, specificity, and accuracy met the acceptance criteria. Conclusion: The present study evidenced the toxicological profile of 18KHT01 polyherbal formulation in mice as well as developed a simple, rapid, and accurate chromatographic method for quality control.