Cytotoxic, acute oral toxicity, genotoxic and mutagenic assessment of the essential oil from fresh leaves of Croton argyrophyllus (Kunth.).

ETHNOPHARMACOLOGICAL RELEVANCE: Croton argyrophyllus Kunth., commonly known as "marmeleiro" or "cassetinga," is widely distributed in the Brazilian Northeast region. Its leaves and flowers are used in traditional medicine as tranquilizers to treat flu and headaches. AIM OF THE STUDY: This study was conducted to determine the chemical composition and toxicological safety of essential oil from C. argyrophyllus leaves using in vitro and in vivo models. MATERIALS AND METHODS: The chemical composition of the essential oil was determined using a gas chromatograph coupled to a mass spectrometer. Cytotoxicity was tested in the HeLa, HT-29, and MCF-7 cell lines derived from human cells (Homo sapiens) and Vero cell lines derived from monkeys (Cercopithecus aethiops) using the MTT method. Acute toxicity, genotoxicity. Mutagenicity tests were performed in Swiss mice (Mus musculus), which were administered essential oil orally in a single dose of 2000 mg/kg by gavage. RESULTS: The main components of the essential oil were p-mentha-2-en-1-ol, α-terpineol, ß-caryophyllene, and ß-elemene. The essential oil exhibited more than 90% cytotoxicity in all cell lines tested. No deaths or behavioral, hematological, or biochemical changes were observed in mice, revealing no acute toxicity. In genotoxic and mutagenic analyses, there was no increase in micronuclei in polychromatic erythrocytes or in the damage and index in the comet assay. CONCLUSIONS: The essential oil was cytotoxic towards the tested cell lines but did not exert toxic effects or promote DNA damage when administered orally at a single dose of 2000 mg/kg in mice.

Evaluation of in silico model predictions for mammalian acute oral toxicity and regulatory application in pesticide hazard and risk assessment.

The United States Environmental Protection Agency (USEPA) uses the lethal dose 50% (LD50) value from in vivo rat acute oral toxicity studies for pesticide product label precautionary statements and environmental risk assessment (RA). The Collaborative Acute Toxicity Modeling Suite (CATMoS) is a quantitative structure-activity relationship (QSAR)-based in silico approach to predict rat acute oral toxicity that has the potential to reduce animal use when registering a new pesticide technical grade active ingredient (TGAI). This analysis compared LD50 values predicted by CATMoS to empirical values from in vivo studies for the TGAIs of 177 conventional pesticides. The accuracy and reliability of the model predictions were assessed relative to the empirical data in terms of USEPA acute oral toxicity categories and discrete LD50 values for each chemical. CATMoS was most reliable at placing pesticide TGAIs in acute toxicity categories III (>500-5000 mg/kg) and IV (>5000 mg/kg), with 88% categorical concordance for 165 chemicals with empirical in vivo LD50 values ≥ 500 mg/kg. When considering an LD50 for RA, CATMoS predictions of 2000 mg/kg and higher were found to agree with empirical values from limit tests (i.e., single, high-dose tests) or definitive results over 2000 mg/kg with few exceptions.

Acute Toxicity Assays with the Artemia salina Model: Assessment of Variables.

The use of the brine shrimp Artemia salina (Leach) in acute toxicity assays has great potential due to its simplicity, low cost and reproducibility. In the current study, some of the variables that can influence the reliability of the assay in terms of test organism survival, were evaluated as part of its implementation in our laboratory. The quality and type of water used, the buffer components and other parameters (salinity, pH and dissolved oxygen level), were all evaluated for optimisation purposes. DMSO (dimethyl sulphoxide) was used as the test substance in the toxicity assay, to evaluate the concentration limits as a solvent in sample preparation. Regarding the buffer salinity, pH and dissolved oxygen level, we found that a 25% to 30% deviation from the standard values did not affect the survival of the nauplii (the first-instar larval stage) under assay conditions. In summary, we corroborate the potential use of this model for the prediction of the toxic potential of substances, to inform future testing strategies.

Acute and Sub-acute Oral Toxicity Assessment of Ferrum phosphoricum in Rats.

BACKGROUND: Ferrum phosphoricum (FP) has been used by traditional medicine practitioners for various ailments since ancient times. However, scientific evidence on the safety of FP is still unavailable. Thus, the current study aimed to investigate the acute and sub-acute oral toxicity of homeopathic FP in experimental rats. METHODS: In an acute toxicity investigation, a single dose of 2,000 µL/kg of FP 6c, 30c and 200c was administered to female Wistar rats, which were monitored for up to 14 days according to the Organization for Economic Cooperation and Development (OECD) guideline 423. For a sub-acute toxicity study, FP 6c, 30c and 200c (200 µL/kg) were administered to male and female rats for 28 days as per the OECD guideline 407. All the animals were observed for mortality, clinical signs and body weight during the study. At the end of the experiment, hematological, biochemical and histopathological assessments were performed. RESULTS: During the acute toxicity study, no mortality was observed in rats administered with FP, and thus the median lethal dose (LD50) was identified as >2,000 µL/kg. In the sub-acute study, no mortality or adverse clinical signs were noticed with FP treatment. Moreover, weekly body weight gain was normal. Hematological and biochemical investigations revealed no abnormalities. Furthermore, histological analysis of FP-treated rats' vital organs revealed no pathological changes. CONCLUSION: Overall, our findings imply that FP 6c, 30c and 200c potencies are safe and do not cause toxicity when given orally to Wistar albino rats for an extended period at a dose of 200 µL/kg.

New supporting data to guide the use of evident toxicity in acute oral toxicity studies (OECD TG 420).

Currently there are three test guidelines (TG) for acute oral toxicity studies of substances or mixtures from the Organisation for Economic Co-operation and Development (OECD). TG 423 and TG 425 use lethality as an endpoint, while TG 420 replaces death with 'evident toxicity', defined as clear signs that exposure to a higher dose would result in death. However, the perceived subjectivity of 'evident toxicity' may be preventing wider use of TG 420. To address this, the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) and the European Partnership for Alternative Approaches to Animal Testing (EPAA) collaborated to provide recommendations on the recognition of 'evident toxicity'. Historical data from acute oral toxicity studies were analysed for clinical signs at the lower dose that could have predicted death at the higher dose. Several signs including ataxia, laboured respiration, and eyes partially closed, alone or in combination, are highly predictive. Others such as lethargy, decreased respiration, and loose faeces have lower but still appreciable positive predictive value (PPV). The data has been used to develop recommendations to promote use of TG 420 and thus reduce the suffering and numbers of animals used in acute oral toxicity studies.

Assessment of cytotoxicity, acute, subacute toxicities and antioxidant activities (in vitro) of Sanghuangporus vaninii crude polysaccharide.

ETHNOPHARMACOLOGY RELEVANCE: Sanghuangporus vaninii (S. vaninii), as a traditional large medicinal fungus, has a history of more than 2000 years in Chinese history and has been widely used to treat female diseases such as vaginal discharge, amenorrhea, and uterine bleeding, and recent pharmacological studies have also found that it has antioxidant, anti-inflammatory, and anti-tumor physiological activity, which has received more and more attention. AIM OF THE STUDY: The objective was to evaluate cytotoxicity and the acute, subacute toxicity, and in vitro antioxidant activity of S. vaninii crude polysaccharide (SVP). MATERIALS AND METHODS: The monosaccharide composition of SVP was determined by HPLC (high-performance liquid chromatography). The cytotoxicity of different concentrations of SVP on three types of cells (HT-22, Kupffer macrophages, HEK293) was assessed using CCk-8. The acute toxicity in vivo was evaluated for 14 days after the administration of SVP (2500,5000, or 10,000 mg/mL). For the evaluation of subacute toxicity, mice were daily treated for 28 days with SVP (2500,5000, or 10,000 mg/mL). In addition, DPPH, hydroxyl radical, and superoxide anion radical were used to evaluate the in vitro antioxidant activity of SVP. RESULTS: SVP was not toxic in all three cell lines tested. In vitro antioxidant tests on the extracts showed that SVP possessed a strong antioxidant capacity in vitro. In the acute study, the no-observed-adverse-effect level (NOAEL) in male and female rats was 10，000 mg/kg body weight. There were also no deaths or severe toxicity associated with SVP in subacute studies. However, SVP treatment had a decreasing effect on body weight in mice of both sexes (2500, 5000, and 10000 mg/kg). At doses (5000 and 10,000 mg/kg), SVP had a reduced effect on food intake in both male and female mice. In addition, there were significant effects on organ coefficients of the liver, lung, and kidney. Hematological analysis showed significantly lower LYM (%) values in mice of both sexes, with significantly lower MCH (pg) values obtained in males (5000 mg/kg and 10000 mg/kg) and higher GRAN (%) values in females. In addition, the RDW-SD (fL) values were significantly lower in the male mice given the highest dose. Biochemical tests showed that there were no significant changes in ALT, AST, TP, and Cr levels after SVP treatment. In histopathological analysis, mild liver toxicity was observed in both female mice treated with 10,000 mg/kg SVP. CONCLUSION: The extract of SVP showed a predominance of polysaccharide compounds, with non-toxic action in vivo. Our approach revealed SVP on the chemical composition and suggests a high margin of safety in the popular use of medicinal fungi. In conclusion, our results suggest that SVP is safe, and can be used as health care products and food.

Safety assessment of European cranberrybush (Viburnum opulus L.) fruit juice: Acute and subacute oral toxicity.

European cranberrybush (ECB) (Viburnum opulus L.) fruits are abundant in phenolic compounds associated with various health benefits. However, the toxicity and safety of ECB juice have not been systematically studied. In the present study, acute and subacute oral toxicities of ECB fruit juice were evaluated on Sprague-Dawley rats and BALB/c mice to establish a toxicity profile. In acute tests, a single administration of 2000 mg/kg body weight of extract to rats exhibited no clinical signs of toxicity or mortality, indicating that the lethal dose (LD50) was over 2000 mg/kg. In subacute tests, repeated administration for 28 days at 0 (control), 500, and 2000 mg/kg doses of extract in mice did not display adverse clinical signs or deaths. However, in the 2000 mg/kg subacute group, platelet counts were significantly high, which correlated with histopathological analyses revealing that ECB extract at 2000 mg/kg was toxic to the kidney, liver, and adipose tissue. The NOAEL value of ECB extract was found as 500 mg/kg/day, but further sub-chronic and chronic toxicity studies are warranted to comprehensively evaluate the long-term safety implications. The study's results emphasize the importance of considering the dosage of dietary supplements containing high levels of phenolic compounds over an extended period to avoid potential cumulative effects from prolonged consumption of high doses.

Assessment of Biological Activities, Acute and Sub-chronic Toxicity of Liang (Gnetum gnemon var. tenerum) Leaves Powder, a Natural Product.

Gnetum gnemon var. tenerum (Gnetaceae) is a shrub plant native to South-East Asia. In Thailand, Liang leaves are commonly consumed in South of Thailand as vegetable. According to literature, they have an antihyperglycemic capacity because of their rich chlorophyll, fiber, and protein. However, there is need to assess the safety since natural food products are not completely devoid of toxicity. This study aimed to assess the biological activities as well as the acute and sub-chronic oral toxicity of Liang leaves powder (LLP). The evaluation of LLP for acute oral toxicity was performed at dose level 2000 mg/kg body weight in Wistar rats while the sub-chronic oral toxicity of LLP was performed at the effective dose (1.47 g/kg) for antihyperglycemic property according to Organisation for Economic Co-operation and Development (OECD)-425. The results showed that LLP demonstrated anti-inflammatory activities. It also showed no clinical signs of toxic effects and mortality in rats throughout 90 d. Thus, LLP could be classified in GHS category 5 which are of relatively low acute toxicity and then the lethal dose, 50% (LD50) cut off at 5000 mg/kg body weight to infinity (∞). Administration of LLP to the experimental rats significantly increased (p < 0.05) the concentration of triglyceride and increased concentration of creatinine as a result of kidney malfunction was also noticed in the experimental rats. Hematological alteration was not noticed in the treated female rats, but red blood cell, hemoglobin and hematocrit concentrations significantly increased in the treated male rats. The study concludes that sub-chronic administration of 1.47 g/kg LLP is relatively safe.

Safety assessment of ethanolic extract of Canarium strictum Roxb. leaves: Acute and subchronic toxicity studies.

Canarium strictum Roxb. (Burseraceae) is a tree distributed in India, China and Thailand. In traditional Ayurvedic medicine, it is used to treat asthma, rheumatism, blood impurities, syphilis, fever, epilepsy and cough. Toxicological information is currently unavailable warrants present research. Ethanol leaf extract obtained by soxhlet extraction was used to investigate its toxicity. The acute toxicity data showed ethanolic leaf extract is safe up to 2000mg/kg dose in female albino mice. There were no behavioral or physiological alterations or gross clinical abnormalities. The ethanolic leaf extract was administered orally to Wistar rats (n=5) of both sexes at a dose of 300, 600 and 1200mg/kg/d for 90 days during the investigation of sub-chronic toxicity. There were no treatment-related deleterious effects on general behavior, body weight, relative organ weight, biochemical and hematological parameters in the sub-chronic trial when evaluated daily/weekly. Organ histopathology revealed no significant abnormalities. Additionally, the ethanolic leaf extract improved rats' cholesterol and metabolic profiles. There is no apparent harm with ethanolic leaf extract treatment for 13 weeks, unless the dosage is quite high. Thus, it implies that the leaves are safer to use as a traditional medicine remedy for a variety of conditions in a wide dose range.

The toxicity assessment of neorudin in cynomolgus monkeys.

In this study, the toxicity effects on circulatory system and respiratory system, and the acute toxicity test of recombinant neorudin (EPR-hirudin, EH) in cynomolgus monkeys were evaluated to provide reference information for clinical studies. Eighteen cynomolgus monkeys were randomly divided into three groups for single intravenous administration of 3, 30 mg/kg EH and normal saline, respectively. The changes of respiratory frequency, respiratory intensity, blood pressure and electrocardiogram before and after administration were recorded. In acute toxicity test, six cynomolgus monkeys were intravenously received EH at a single dose of 171, 257, 385, 578, 867 and 1300 mg/kg respectively. The vital signs, hematology, serum biochemistry, coagulation indexes and electrocardiogram indexes of the animals were determined before administration and on the 7th and 14th day after administration. As the results showed that there were no significant abnormal changes in respiratory frequency, respiratory intensity, blood pressure or electrocardiogram in cynomolgus monkeys after receiving EH at 3 mg/kg and 30 mg/kg, and there was no statistical difference between the treated groups and normal saline group. In the acute toxicity test, no significant abnormalities were observed in vital signs, hematology, serum biochemistry, coagulation indexes and electrocardiogram indexes of six cynomolgus monkeys at day 7 and 14 after EH administration. Furthermore, autopsies of all cynomolgus monkeys showed no abnormalities. The results of toxicokinetics showed that AUClast of the drug increased in proportion to the EH dose in the range of 171-578 mg/kg, and increased in over proportion to the EH dose in the range of 578-1300 mg/kg. The variation of Cmax was basically consistent with AUClast. In a sum, A single intravenous injection of 3 and 30 mg/kg of EH did not affect the circulatory system and respiratory system in cynomolgus monkeys and the maximum tolerated dose of EH in cynomolgus monkey is over 1300 mg/kg (equivalent to 619-1300 times of the proposed clinical equivalent dose).

Environmental safety of second and third generation bioplastics in the context of the circular economy.

Bioplastics derived from organic materials other than crude oil are often suggested as sustainable solutions for tackling end-of-life plastic waste, but little is known of their ecotoxicity to aquatic species. Here, we investigated the ecotoxicity of second and third generation bioplastics toward the freshwater zooplankton Daphnia magna. In acute toxicity tests (48 h), survival was impacted at high concentrations (g.L-1 range), within the range of salinity-induced toxicity. Macroalgae-derived bioplastic induced hormetic responses under chronic exposure (21 d). Most biological traits were enhanced from 0.06 to 0.25 g.L-1 (reproduction rate, body length, width, apical spine, protein concentration), while most of these traits returned to controls level at 0.5 g.L-1. Phenol-oxidase activity, indicative of immune function, was enhanced only at the lowest concentration (0.06 g.L-1). We hypothesise these suggested health benefits were due to assimilation of carbon derived from the macroalgae-based bioplastic as food. Polymer identity was confirmed by infra-red spectroscopy. Chemical analysis of each bioplastic revealed low metal abundance whilst non target exploration of organic compounds revealed trace amounts of phthalates and flame retardants. The macroalgae-bioplastic disintegrated completely in compost and biodegraded up to 86 % in aqueous medium. All bioplastics acidified the test medium. In conclusion, the tested bioplastics were classified as environmentally safe. Nonetheless, a reasonable end-of-life management of these safer-by-design materials is advised to ensure the absence of harmful effects at high concentrations, depending on the receiving environment.

Assessment of sub-chronic oral toxicity of Nityanand Rasa: An ayurvedic herbo-metallic formulation.

ETHNOPHARMACOLOGICAL RELEVANCE: Nityananda Rasa (NR) is an ayurvedic herbo-metallic formulation used to treat gout, obesity, hypothyroidism, elephantiasis, and other diseases. However, its safety is a concern owing to the use of heavy metals like mercury and arsenic. AIM OF THE STUDY: To study the sub-chronic oral toxicity of NR on albino wistar rats for safety evaluation. MATERIALS AND METHODS: The male and female albino wistar rats were administered a daily dose of 30 (low), 300 (medium) and 600 (high) mg/kg BW/day of NR for 90-day period. The body weight and feed consumption were monitored once a week. After 90 days, blood and vital organs were harvested for genotoxicity, hematology, biochemistry, histopathology, gene expression and the biodistribution analysis. RESULTS: There was no mortality or severe behavioural changes observed in rats. Significant changes in biochemical enzyme levels were seen at medium and high doses of NR i. e. 300 and 600 mg/kg BW/day respectively. No hematological changes were observed. Mild histopathological changes seen at high dose of NR which were found in concurrence with the biochemical alterations in liver and brain. There was mild genotoxicity and no detectable level of mercury but significant arsenic level in blood at high dose. Gene expression was mildly affected. CONCLUSIONS: NR induced moderate toxic effects at high dose but can be considered safe at therapeutic doses.

Phytochemical Analysis and Toxicity Assessment of Bouea Macrophylla Yoghurt.

The Bouea macrophylla fruit is native to Malaysia and is known for its many beneficial effects on one's health. Probiotics are well-known for their roles as anti-inflammatory, antioxidant, and anti-tumour properties due to their widespread use. As a result, the purpose of this study was to incorporate the ethanolic extract of Bouea macrophylla into yoghurt and then assess the rodents for any toxicological effects. According to the findings of the nutritional analysis, each 100 mL serving of the newly formulated yoghurt contains 3.29 g of fat, 5.79 g of carbohydrates, 2.92 g of total protein, and 2.72 g of sugar. The ability of the newly developed yoghurt to stimulate the growth of Lactobacilli was demonstrated by the fact that the peak intensity of Lactobacillus species was measured at 1.2 × 106 CFU/g while the titratable acidity of the lactic acid was measured at 0.599 CFU/g. In order to carry out the toxicological evaluation, forty-eight male Sprague Dawley (SD) rats were utilized. Oral administration of single doses of 2000 mg/kg over the course of 14 days was used for the study of acute toxicity. Subacute toxicity was studied by giving animals Bouea macrophylla yoghurt (BMY) at repeated doses of 50, 250, 500, and 1000 mg/kg/day over a period of 28 days, while the control group was given normal saline. The results of the acute toxicity test revealed that rats treated with increasing doses up to a maximum of 2000 mg/kg exhibited no signs of toxicity. After an additional 14 days without treatment, acute toxicity of a single dose (2000 mg/kg) of BMY did not show any treatment-related toxicity in any of the rats that were observed. According to the data from the subacute toxicity study, there were no differences between the treated groups and the control groups in terms of food and water intake, body weight, plasma biochemistry (AST, ALT, ALP, and creatinine), haematological products, or organ weights. The architecture of the liver, heart, and kidney were all found to be normal upon histological examination. This indicates that oral consumption of BMY did not result in any negative effects being manifested in the rodents.

A toxicological evaluation for safety assessment of ruthenium-based diosmetin complex in rats.

The flavonoid-based organometallic complexes have been identified as novel bioactive compounds with enhanced pharmacological and therapeutic activity. In this study, the ruthenium-p-cymene diosmetin complex was synthesized, characterized, and investigated for toxicological profiling through different toxicological and genotoxicological studies which include acute and sub-acute toxicity, chromosomal aberration, and bone marrow micronucleus study. The acute oral toxicity study demonstrated the LD50 dose of the complex at 500 mg/kg body weight which further instigated the sub-acute doses i.e. 50, 100, and 200 mg/kg. The histopathological analysis demonstrated that the 400 mg/kg dose was associated with severe toxicological incidences of the vital organs (liver, kidney, pancreas, testis, and stomach) except the ovary with increased levels of ALP, AST, ALT, and WBC count. However, 50, 100, and 200 mg/kg doses did not show any toxicological alteration and maintained the normal levels of hematological and serum biochemical parameters. The genotoxicological assessment of the complex depicted no such genetic damage or mutagenicity in any complex treated groups. In conclusion, the 50, 100, and 200 mg/kg doses were determined as therapeutic dose of the novel ruthenium-p-cymene diosmetin complex without any genotoxic and mutagenic potential which can be further implemented in the investigation of various pharmacological and therapeutic interventions.

Safety assessment of oil extracted from lacquer (Toxicodendron vernicifluum (Stokes) F.A. Barkley) seed: acute and subchronic toxicity studies in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Toxicodendron vernicifluum (Stokes) F.A. Barkley (RVS) is an economic tree species and widely distributed in East Asia. Wood parts and raw lacquers of RVS have been used in coatings, herbal medicines or food supplements, and the leaves, flowers, roots, and fruits of RVS are also widely used in medicine traditionally. Lacquer seed oil (LSO) has potential health benefits and has not previously been evaluated for safety. AIM OF THE STUDY: The aim of the present study was to investigate the toxicological potential of LSO by acute and subchronic toxicity tests. MATERIALS AND METHODS: The characterization of fatty acids of the LSO was carried out by gas chromatography. In the acute toxicity study, LSO was administered at single doses of 5000 or 10000 mg/kg by oral gavage. The subchronic toxicity study was conducted by daily oral administration of LSO at doses of 1250, 2500 and 5000 mg/kg/day for 30 consecutive days. The animals were evaluated for clinical observations, body weight, organ weight, feed consumption, biochemical and hematological parameters, and liver, lung, and kidney histology. RESULTS: There were no mortality and toxic changes were observed in acute toxicity study. The results of subchronic toxicity showed no toxicologically significant changes in clinical observations, body weight, organ weight, biochemical or hematological parameters. Histopathologic results indicated slight hepatic steatosis and inflammatory infiltration in the rats of 5000 mg/kg/day LSO treated group. However, the histopathologic observation was not confirmed by hepatic biochemical analysis. CONCLUSIONS: These results suggested that the LD50 of LSO is over 10000 mg/kg and LSO is non-toxic for SD rats in acute toxicity study. The no observed adverse effect level (NOAEL) of LSO in rats is considered to be 5000 mg/kg/day, and liver is the potential target organ of LSO for 30-day subchronic toxicity study.

Development a high-throughput zebrafish embryo acute toxicity testing method based on OECD TG 236.

The Organization for Economic Co-operation and Development (OECD）Test Guideline (TG) 236 for zebrafish embryo acute toxicity testing was adopted for chemical toxicity assessment in 2013. Due to the increasing demand for prediction and evaluation of the acute toxicity using zebrafish embryos, we developed a method based on OECD 236 test guideline with the aim to improve the testing efficiency. We used 4-128 cell stage zebrafish embryos and performed an exposure assay in a 96-well microtiter plate, observing the lethality endpoints of embryos at 48-h postexposure. A total of 32 chemicals (two batches) were used in the comparison study. Our results indicated that the logarithmic LC50 (half lethal concentration) obtained by the modified method exhibited good correlation with that obtained by the OECD 236 testing method, and the R2 of the linear regression analysis was 0.9717 (0.9621 and 0.9936 for the two batches, respectively). Additionally, the intra- and inter-laboratory coefficient of variation (CVs) for the LC50 from the testing chemicals (17 chemicals in second batch) was less than 30%, except for CuSO4. Therefore, the developed method was less time-consuming and demonstrated a higher throughput for toxicity testing compared to the prior method. We argue the developed method could be used as an additional choice for high-throughput zebrafish embryo acute toxicity test.

Secondary metabolite contents and safety assessment study of the aqueous extract from the Algerian Echium trygorrhizum Pomel roots.

ETHNOPHARMACOLOGICAL RELEVANCE: Species of Echium trygorrhizum Pomel, belonging to the Boraginaceae family, grow wild in North Africa. This plant is used in traditional Algerian medicine for the treatment of Diabetes, Jaundice and Tonsillitis. AIM OF THE STUDY: To our knowledge, no work has been done on the phytochemistry and biological activities of this plant. Moreover, no study has yet corroborated that the use of this plant is safe. Therefore, the present study was carried out to gather information on the various medicinal uses of this plant and to evaluate the total phenolic content and assess its safety after acute and sub-acute toxicity tests with rodents (mice and rats). MATERIAL AND METHODS: An ethnopharmacological survey was carried out using a direct questionnaire. It included some questions relating to the plant. The content of polyphénols, flavonoids and tannins present in the aqueous extract was determined by colorimetric methods. In the acute toxicity tests, three groups each containing five males and five females Albino mice were formed. The control group received water, while the test groups received different doses of aqueous extract (2000 and 5000) mg/kg body weight. In the subacute toxicity study, different doses (250, 500 and 1000 mg/kg) of the aqueous extract were administered to male and female rats for 28 consecutive days. Hematological analysis, biochemical parameters estimation and histopathological examination have been determined at the end of the treatment. RESULTS: Data from the ethnopharmacological survey showed that 25, 31% of people used this plant in Algeria as a traditional medicine for the treatment of Jaundice (100%), Diabetes (28, 33%) and Tonsillitis (10%). Results also revealed that aqueous extract contains high amounts of polyphenols, flavonoids and tannins. The extract did not cause any toxicity during the 14-days observation period after a single dose of 5000 mg/kg was administered to mice, with LD50 values greater than 5000 mg/kg. In a further step, the administration of the aqueous extract at all dose levels (250, 500, and 1000 mg/kg) to male and female rats during the 28-days study did not result in any deaths or behavioral changes, and there were no changes in body weight, relative organ weights, or food consumption as a result of this experiment. However, male rats showed a significant difference in relative liver weight in the high dose group (1000 mg/kg) (p < 0.05) and the satellite group (p < 0.01). In female rats, liver weight significantly increased only in the satellite group compared with the control group (p < 0.01). The results showed an increase in red blood cells (RBC), hemoglobin (Hb), and hematocrit (Hct) in the female groups. However, a clear decrease was observed in plasma ALP enzyme activities in females treated with the doses of 250 and 1000 mg/kg (p < 0.05) after the treatment periods. CONCLUSION: The study indicated that the single dose of 5 g/kg AQE can be considered relatively safe as it did not cause death or any signs of toxicity in mice. Repeated oral administration of AQE at doses below 250 g/kg/day for 28 consecutive days can be considered relatively safe.

Prediction of acute oral toxicity using the Hansen solubility parameter.

Acute oral toxicity is primarily determined using animal testing, as stated in the Organization for Economic Cooperation and Development (OECD) Test Guideline (TG) 420, 423, and 425. Currently, regarding animal welfare, few alternatives to animal testing such as in vitro approaches have been evaluated. Therefore, in this preliminary study examining a new method to determine acute oral toxicity, we investigated whether UN Globally Harmonized System all categories can be predicted using the Hansen solubility parameter (HSP). In particular, Hansen spheres were produced based on oral toxicity information of the test substances and their HSP values, and the respective parameters were identified. A comparison of these potential parameters with the HSP value of each test substance showed an accuracy of 84.1% (53/63), 10.0% (3/30) false negatives, and 21.2% (7/33) false positives. By comparing the HSP of the resulting potential parameters with a test substance, it is possible to predict acute oral toxicity with high accuracy.

Safety assessment of MPTA: An oral acute and 90-day sub-chronic toxicity study in Sprague-Dawley rats.

MPTA is a novel extract product derived from Macleaya cordata (Willd.) R. Br., which has good anti-inflammatory and antioxidant activity. The aim of this study was to investigate the acute oral toxicity and 90-day sub-chronic oral toxicity of MPTA. In the acute toxicity study, 50 SD rats of both sexes were randomly divided into 5 groups and dosed in a gradient from 197.53 mg/kg to 1000.00 mg/kg bw. Toxic effects were observed up to 14 days and LD50 was calculated. In a subchronic toxicity test, male and female SD rats were orally dosed repeatedly with 96.40, 19.28, 3.86 mg/kg bw of MPTA for 90 days. In addition, a control group was set up in the subchronic study. The acute toxicity test showed that the oral LD50 of MPTA was 481.99 mg/kg with a 95% confidence interval of 404.24-574.70 mg/kg. MPTA did not appear to induce toxic effects in the longer term in terms of food and water consumption, weight gain, haematological and clinical biochemical parameters and pathological examination. The first data on the potential toxicity of MPTA was provided to highlight the safety of short-term to longer-term oral administration of MPTA, and the experimental results yield and establish a NOEAL of 96.40 mg/kg/d for MPTA.

Safety Evaluation of Oleoresin-Based Turmeric Formulation: Assessment of Genotoxicity and Acute and Subchronic Oral Toxicity.

Turmeric rhizome (Curcuma longa L.) has been used without concern for safety as a culinary spice and traditional medicine under the ancient Ayurvedic medicinal system of India dating back nearly 4000 years. This preclinical safety evaluation was done to determine the safety of an oleoresin-based turmeric extract (CURCUGEN®). Guidelines from the Organization for Economic Co-operation and Development (OECD) directed the assessment of safety for the in vitro and in vivo application of CURCUGEN®. Safety of the herbal medicine was evaluated through the toxicological assessment of acute, oral, and 90-day repeated dosing, genotoxicity, and mutagenicity study. Genotoxicity tests included the in vitro bacterial reverse mutation test, chromosomal aberration test, and in vivo micronucleus test. The single dose of CURCUGEN® administered orally (gavage) to Sprague-Dawley (SD) rats resulted in a LD50 of >5000 mg/kg body weight. The subchronic assessment of CURCUGEN®, as administered to SD rats over 90 days resulted in a no observed adverse effect level (NOAEL) of 2000 mg/kg body weight/day. CURCUGEN® did not elicit any genotoxic or clastogenic effect in genotoxicity tests. The battery of safety studies carried out demonstrated that CURCUGEN® showed no evidence of general toxicity or genotoxicity.