Subacute Oral Toxicity Research Articles

Acute and Sub-Acute Toxicity of Spray-Dried Extract of Tribulus Terrestris

The use of herbal medicines has tremendously grown over the past few years. Owing to their safety, they are more preferred over conventional allopathic medicine. Plant-based therapies have significantly improved and protected human health. Additionally, dry powdered herbal medications are more effective than liquid extracts. The numerous claims are not backed up by any data that can be verified. It is the first time that the activity of Tribulus Terrestris spray-dried extract has been linked to acute and suba-cute oral toxicity. The objective of the current work was to investigate the acute and sub-acute toxicity of spray-dried extract (SDE) of Tribulus terrestris. For acute toxicity, two different doses, i.e., 2000 and 5000 mg/kg, were given to the female Wistar rats. Only one dose was administered and each animal was observed for any signs of mortality along with behavioral changes and physical symptoms for 2 weeks. Multiple doses of the SDE were given to the Wistar (both genders) rats for 28 days in the sub-acute toxicity. Animals of both the genders received 500, 1000, and 1500 mg/kg/day SDE doses for 28 days consecutively. Blood samples were obtained for biochemical and hematological analysis on the 29th day of the experiment, following which subjects were sacrificed for the histopathological examination. The results of acute toxicity revealed no signs of morbidity and mortality in rats. Similar results were recorded in the sub-acute toxicity. Mild variations were observed at 1000 and 1500 mg/kg/day, however results were non-significant. The SDE of Tribulus terrestris did not show any significant harmful effects in the treated animals after 14 and 28 days of treatment. Therefore, SDE of Tribulus terrestris can be safely employed in pharmaceutical formulations for therapeutic use, addressing further safety in chronic and sub-chronic study.

A pilot study of toxicological safety evaluation in acute and subacute (28-day) oral assessments of kenaf seed beverage in rats

Kenaf (Hibiscus cannabinus L.) seed is a non-conventional edible oilseed that can be valorized into various food products. There is a recent discovery of kenaf seed beverage (KSB) potential as a novel plant-based beverage. KSB had less crude protein than soybean (SB)but more carbohydrate, magnesium, and phosphorus contents.Levels of crude fat, phytates, oxalates, total saponins, and lipid peroxidability in KSB were lower than SB. Sugar content between KSB and SB were comparable, while antioxidant properties of KSB were superior. Ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) analysis detected gluconic acid, citric acid, palmitic acid, oleic acid, and 13-hydroxyoctadecadienoic acid in both KSB and SB. Considering its novelty, acute and subacute oral toxicity assessments in male Sprague Dawley rats were conducted. The acute toxicity assessment was performed at a single dose of 9.2 ml/kg body weight of KSB. In the following subacute toxicity assessment, different groups of rats consumed different doses of KSB (3.1, 6.1, and 9.2 ml/kg body weight) daily for 28 days. Rats presented normal behavioral and physiological states in both toxicity studies. Growth, food and water intakes, organ weight, and hematological parameters were unaffected. No mortality was reported. Several alterations in serum biochemical parameters were within the normal range, and unassociated with histopathological changes. The oral lethal dose (LD50) and the no-observed-adverse-effect-level (NOAEL) of KSB in rats was greater than 9.2 ml/kg (=1533 mg/kg) body weight. Interestingly, KSB exhibited comparable effects with soybean beverage (SB) on high-density lipoprotein cholesterol and triglycerides which worth further research Follow-up toxicity assessments in animals and human trials are also recommended to ascertain its long term safety.

Preclinical Safety Assessment of Chemically Cross-Linked Modified Mandua Starch: Acute and Sub-Acute Oral Toxicity Studies in Swiss Albino Mice.

In the present era, 28 days of repeated-dose-toxicity study following the Organization for Economic Cooperation and Development (OECD) guidelines 407 is compulsory for every drug to go through phase 1 clinical trials. The increasing demand for high-resistant starch containing nutraceuticals and the applicability of modified starch in development of targeted drug delivery inspired us to investigate the toxic profile of mandua starch chemically cross-linked by epichlorohydrin and compare it with alkali-isolated starch in healthy adult Swiss albino mice, which can be the first step for exploring the use of epichlorohydrin-cross-linked mandua starch (ECC-MS) as a pharmaceutical excipient. Histopathological examinations of the kidney and liver did not expose noteworthy abnormalities in the treated mice. There were no clinical and mortality symptoms of toxicity observed during the repeated-dose-toxicity study. The oral consumption of ECC-MS did not pose any harm as it was neither lethal nor had any harmful hematological, biochemical, psychological, anatomical, and behavioral effects. The use of ECC-MS and alkali-isolated mandua starch (AMS) was found safe at a dose of 2000 mg/kg body weight in the acute toxicity study and at doses of 2000, 1500, and 1000 mg/kg body weight in the sub-acute toxicity study as no detrimental effects were observed after oral administration in mice for 14 and 28 days, respectively.

Toxicity evaluation induced by single and 28-days repeated exposure of withametelin and daturaolone in Sprague Dawley rats.

Safe preclinical dose determination is predictive of human toxicity and can have a profound impact on the overall progress of the compound in early drug discovery process. In this respect, current study sought to investigate for the first time the acute and subacute oral toxicity of two pharmacologically active natural compounds i.e., withametelin and daturaolone in Sprague Dawley rats following OECD guideline 420 and 407, respectively. As per acute toxicity studies, withametelin and daturaolone were characterized as Globally Harmonized System (GHS) category 4 and 5 compounds, respectively. Sub-acute daily dose of withametelin was 5, 2.5, and 1.25 mg/kg but, for daturaolone, it was 10, 5, and 2.5 mg/kg. High dose (5 and 2.5 mg/kg) withametelin groups showed dose dependent changes in the general, hematological, biochemical and histopathological parameters in both sexes, the most prominent being hyperthyroidism while no toxicity was observed at lower doses (1.25 and 0.75 mg/kg), No Observable Adverse Effect Level (NOAEL) being 1.25 mg/kg. Daturaolone was comparatively safer and showed dose dependent significant changes in hepatic enzyme (Alanine Transaminase), bilirubin, creatinine, and glucose levels while histological changes in testes were also observed. Lower doses (5, 2.5, and 1.25 mg/kg) of daturaolone showed no significant toxic effects and 5 mg/kg was declared as its NOAEL. Depending upon our findings, starting effective oral dose levels of 1.25 mg/kg/day for withametelin and 5 mg/kg/day for daturaolone are proposed for repeated dose (up to 28 days) preclinical pharmacological evaluation models. Long term studies with more behavioral, biochemical, histopathological and hormonal parameters are proposed to strengthen the findings.

Green and proficient process for industrial-scale preparation of Gymnema sylvestre standardized-extract enriched with Gymnemic acids through polymer-matrix-adsorption to reduce hyperglycemia

Gymnema sylvestre is an important medicinal plant, but a method for preparing standardized extract enriched with biomarkers is not available. The practiced crude hydro-alcoholic extracts of G. sylvestre contained a few percentage of gymnemic acids (GAs). The enrichment of GAs in preprared extracts is essential from a pharmacology point of view. Presently, a scale-up process has been developed for preparing G. sylvestre standardized extract (Gym-EEXT). Crude extract (Gym-Crude) was prepared in 10-kg batches to obtain a yield of 268.1 mg/g, but GAs was merely 9 mg/g (GA-IV 6.7 mg/g, and GA-XVII 2.3 mg/g) in 30 % ethanol–water solvent at 50 °C in 5 h. Further, Gym-Crude was processed through polymer-matrix-adsorption techniques for getting Gym-EEXT (150.9 mg/g) with improved percentage of GAs (>10 %) comprised of deacylgymnemic acid (DGA: 8.9 mg/g), gymnemagenin (2.3 mg/g), GA-IV (78.6 mg/g), and GA-XVII (10.2 mg/g). For safety aspects, the pesticide residues, solvent residues, heavy metal, aflatoxins, and mutagenicity in Gym-EEXT were below the detection limits.Hence, Gym-EEXT was enriched with GAs, and fully safe with compliance according to USP-561.An insignificant effect of Gym-EEXT on vitals of experimental mice was observed in acute and sub-acute oral-toxicity experiments.Gym-EEXT significantly improved the blood glucose metabolism, and hence, it potentially reduces hyperglycaemia. Unique chromatographic protocols were developed for the isolation of biomarkers on a 1-kg scale from Gym-Crude with yield of gymnemagenin (5.5 %), and DGA (4.6 %). Finally, a novel, scientifically validated, and the scale-up process has been developed for the preparation of Gym-EEXT along with an exclusive protocol for isolating its biomarkers.

Acute and subacute oral toxicity of artemisinin-hydroxychloroquine sulfate tablets in beagle dogs

Artemisinin-hydroxychloroquine sulfate tablets (AH) are regarded as a relatively inexpensive and novel combination therapy for the treatment of various forms of malaria, particularly aminoquinoline drugs-resistant strains of Plasmodium falciparum. Our aim was to conduct acute and subacute oral toxicity studies in non-rodents to obtain more nonclinical data on the safety of AH. Acute toxicity evaluation was performed in beagle dogs at single doses of 230, 530, 790, 1180, 2660, and 5000 mg/kg. Beagle dogs at doses of 0, 56, 84, and 126 mg/kg were used to assess subacute toxicity for 14 days. The approximate lethal dose range for acute oral administration of AH in dogs is found to be 790–1180 mg/kg, and toxic symptoms prior to death include gait instability, limb weakness, mental fatigue, tachypnea, and convulsion. Repeated doses of AH in dogs caused vomiting, soft feces, decreased activity, anorexia, and splenic red pulp vacuolation. Of note, AH could reduce body weight gain and prolong the QTc interval of individual dogs. Therefore, the no-observed-adverse-effect level (NOAEL) and lowest-observed-adverse-effect level (LOAEL) of oral administration of AH for 14 days in dogs are determined to be 84 mg/kg and 126 mg/kg, respectively.

Acute and subacute oral toxicity evaluation of Antarctic krill protein in Kunming mice

Antarctic krill (Euphausia superba) protein is widely acknowledged as a potential animal protein source due to its large biomass with excellent nutritional and utilisation properties. However, safety assessments of Antarctic krill protein (AKP) are highly warranted before its use as human food. The present work thus assessed the safety of AKP in a Kunming mice model through acute toxicity and a 28-day feeding study, where the Kunming mice were fed with AKP or control diets. In the acute toxicity study, a single oral dose of 10 g/kg bodyweight (BW) AKP caused no death or abnormal effects in male and female mice, and the bodyweight gain remained within the normal range. In the repeated dose 28-day oral toxicity study, AKP was orally administered to Kunming mice at the doses of 2.5, 5.0, and 10.0 g/kg BW/day for 28 days. The absolute and relative liver weight gained was only observed in the mice administered with high-dose of AKP. However, this increase was incidental as no weight gain or histopathological alterations were observed in the main groups. These findings were consistent with the normal background lesions in the clinically normal mice used in the present work, which were considered spontaneous and/or incidental in nature and unrelated to the treatment. These results demonstrated that AKP did not exert significant acute and subacute toxicity upon oral administration to Kunming mice.

Acute and Subacute Oral Toxicity studies on Aquatic plant Trapa natans L. using a Rat Model

Trapa natans L. is annual aquatic plant generally kwon as Water caltrp, Water chest nut belonging to the Trapaceae or Lytraceae family. Trapa natans L. is use for the treatment of wide no of disease with out proper toxicity study. The present study was designed to evaluate the acute and sub-acute toxicities of Trapa natans L following the OECD guidelines. In acute oral toxicity, the plant extract was administered orally at the doses of 300 mg/kg, 1000 mg/kg, 2000 mg/kg, and 4000 mg/kg. After the dosing, all animals were kept in close observation for 14 days. In the case of the subacute toxicity study, the total number of animals was divided into four groups, each having ten animals, 5 males, and 5 females in each group. Group I was represented as the control, administered orally with aqueous tween 80 solution 2% (v/v), and group II to group IV received plant extract of doses 400, 600, and 1000 mg/kg, respectively, once a day for 28 days. From the acute and subacute toxicity studies, no sign of toxicity was observed. In the sub-acute toxicity study, no major changes were noted on the biochemical parameters and hematological analysis. The experimental results of the toxicity studies suggested that the LD50 values of the Trapa natans L extract were more than 4000mg/kg and thus, the drug was found to be absolutely safe and nontoxic.

Acute and sub-acute oral toxicity assessment of 5-hydroxy-1,4-naphthoquinone in mice

5-hydroxy-1,4-naphthoquinone (5NQ) or juglone is a bioactive molecule found in walnuts and has shown therapeutic effects in various disease models. Limited information is available regarding the toxicity of 5NQ, thereby limiting the clinical development of this drug. In the present study, oral acute (50, 300 and 2000 mg/kg) and sub-acute toxicity (5, 15 and 50 mg/kg) was assessed in mice to evaluate the safety of 5NQ. The acute toxicity study identified 118 mg/kg as the point-of-departure dose (POD) for single oral administration of 5NQ using benchmark dose modeling (BMD). Repeated administration of 5NQ at doses of 15 and 50 mg/kg/day caused reduction in food consumption and body weight of mice along with alterations in liver and renal function. Histopathological assessment revealed significant damage to hepatic and renal tissues at all doses in the acute toxicity study, and at higher doses of 15 and 50 mg/kg in the sub-acute toxicity study. We observed dose dependent mortality in sub-acute toxicity study and the no observed adverse effect level (NOAEL) was established as < 5 mg/kg/day. Modeling the survival response in sub-acute toxicity study identified 1.74 mg/kg/day as the POD for repeated administration of 5NQ. Serum levels of aspartate aminotransferase (AST) were most sensitive to 5NQ administration with a lower limit of BMD interval (BMDL) of 1.1 × 10−3 mg/kg/day. The benchmark doses reported in the study can be further used to determine a reference dose of 5NQ for human risk assessment.

Complete assessment of methanolic extract of Euphorbia tirucalli L. in wistar rat model by subacute oral toxicity

Complete assessment of methanolic extract of Euphorbia tirucalli L. in wistar rat model by subacute oral toxicity

Evaluation of the Acute and Sub-Acute Oral Toxicity of Jaranol in Kunming Mice.

Background: Jaranol has shown a wide range of pharmacological activities; however, no study has yet examined in vivo toxicity. The study aimed to investigate the oral acute and sub-acute toxicity of jaranol in mice. Methods: The acute toxicity was determined by a single oral dose of jaranol (2000 mg/kg). Therein animal behaviour and mortality rate were observed for 14 days. The jaranol (50, 100 and 200 mg/kg BW·d−1) was given by gavage for 28 days daily in the sub-acute study. The mouse body weight (BW), organ weight, food, water intake, biochemical, haematological parameters, and histopathology were studied in acute and sub-acute toxicity. Results: During the acute toxicity test, a single oral dose (2000 mg/kg) jaranol did not cause significant alteration in majority of the hematological indices. However, jaranol decreased the level of serum alanine aminotransferase and aspartate aminotransferase. Those results showed that the oral lethal dose 50 (LD50) of jaranol was higher than 2000 mg/kg BW, regardless of sex. In repeated daily oral doses (50, 100 and 200 mg/kg BW·d−1), no mortality was recorded in the various experimental groups. The jaranol reduced body weight gain (200 mg/kg BW·d−1), the relative spleen weight (all doses) and serum alanine aminotransferase activity (200 mg/kg BW·d−1). On the other hand, jaranol significantly elevated red blood cell count (100 and 200 mg/kg BW·d−1) and serum creatinine levels (200 mg/kg BW·d−1). Histological study revealed that spleen bleeding was identified in 200 mg/kg jaranol-treated mice. Conclusion: Jaranol was relatively safe in Kunming Mice when repetitively administered orally in small doses for a prolonged period of time. We recommend more chronic toxicity studies and clinical trials on jaranol to ensure that its use is free of potential toxicity to humans.

Preliminary Toxicity Studies of Methanol Stem Bark Extract of Lannea acida A. Rich (Anacardiaceae) in Wistar Rats

Background: Despite the use of Lannea acida A. Rich for the treatment of many ailments, its safety profile has not been well-elucidated. We aim to determine the acute and subacute oral toxicity of methanol extract of stem bark of L. acida on Wistar rats. Materials and methods: For acute toxicity test, adult female Wistar rats (n = 5) received a single oral dose of 3000 mg/kg of methanol extract of stem back of L. acida by gavage at interval of 48 hours, and were observed for general behavior, adverse effects and mortality for 14 days. For subacute toxicity, 24 adult Wistar rats were divided into four groups (n = 6/group; male = female, 1:1). Group one received distilled water at th 10 ml/kg body weight while groups two, three and four received 250, 500 and 1000 mg/kg body weight of extract orally for 28 days. On the 29 day, rats were euthanized and morphological, biochemical, and hematological parameters determined. Results: Acute oral dose of 3000 mg/kg body weight did not produce any sign of toxicity or mortality in Wistar rats. Subacute treatment showed a decrease in the relative organ weight percent of the kidney at a dose of 1000 mg/kg body weight (P &lt; 0.05); a decrease in the serum potassium across all the doses (P &lt; 0.05), and a rise in serum alanine transaminase at 500 mg/kg (P &lt; 0.05). Histology showed varying degrees of hepatic sinusoidal vascular congestion, peri-portal inflammation; as well as tubulointerstitial inflammation and glomerular necrosis at doses of 250, 500 and 1000 mg/kg. Conclusion: The acute oral median lethal dose (LD ) of methanol extract of the stem bark of L. acida is greater than 3000 mg/kg body weight. 50 Twenty-eight day daily oral doses of more than 250 mg/kg may have nephrotoxic and hepatotoxic effects in Wistar rats. Keywords: Lannea acida, Wistar, toxicity, acute, sub-acute

Evaluation of Rhodojaponin III from Rhododendron molle G. Don on oral antinociceptive activity, mechanism of action, and subacute toxicity in rodents

Ethnopharmacological relevanceIn Chinese traditional medicine, Rhododendron molle G. Don is a recognized herb to ease pain. Rhodojaponin III (RJ-III) has been identified as the main pharmacological activity and toxic component of the herb; however, oral antinociception and mechanism of RJ-III have not yet been investigated.Aim of the study: The significance of this study is to evaluate the effects of RJ-III on nociceptive and neuropathic pain, and to preliminarily explore the underlying mechanisms and subacute toxicity. Materials and methodsThe antinociception of RJ-III was evaluated by hot plate, tail-immersion, acetic acid writhing, formalin test and chronic constriction injury (CCI) model in rodents. An experimental validation was conducted using whole-cell patch clamp technique based on the most likely mechanisms of action after screening and prediction by molecular docking study. In addition, the oral subacute toxicity of RJ-III was assessed. ResultsBehavioral experiments showed that RJ-III (0.20 mg/kg) reduced the latency of the nociceptive response in the hot plate and tail-immersion tests. Acetic acid and formalin-induced pain were significantly inhibited by RJ-III (0.10 and 0.05 mg/kg, respectively). Furthermore, 0.30 mg/kg of RJ-III improved hyperalgesia in the CCI-induced rats. Based on molecular docking results, electrophysiological experiments were used to demonstrate mild inhibition of voltage-gated sodium channel-related subtypes. Additionally, oral subacute toxicity that may cause leukopenia and abnormal liver function requires further attention in subsequent studies. ConclusionRJ-III mildly blocks voltage-gated sodium channel to inhibit nociceptive pain and peripheral neuralgia, but 0.375 mg/kg and above may cause side effect after long-term oral administration.

English

Erythrina sacleuxii, one of five Erythrina species found in Kenya, is a stiffly multi-branched deciduous tree reported to have antiplasmodial, antifungal, anti-bacterial and anti-cancer activities. This study aimed at evaluating acute and sub-acute oral toxicity of E. sacleuxii roots ethanolic extract. Phytochemical analysis was conducted on Fourier transform infrared spectroscopy and Gas Chromatography-Mass Spectrometer. For acute and sub-acute toxicity studies 16 male rats n=4/group were used. The control group received 2% DMSO while treatment groups received 400, 1200 and 2400 mg/kg single dose (acute toxicity assay) and repeated for 14 days in subacute toxicity assay. Histopathological, serum biochemistry and hematological tests were analyzed. The results of FT-IR analysis recorded 12 dominant peaks, confirming availability of N-H, C-N, -C=C-, C-H, S=O, C-O and C-C functional groups. GC-MS analysis revealed presence of 17 bioactive phytochemicals mainly terpenoids and phenols. No toxic effect, organ toxicity or death was recorded in acute toxicity assay. In the subacute toxicity investigation, rats&#39; behavior, gross pathology, and hematological parameters showed no significant changes. Biochemical analysis indicated an increase in alkaline phosphatase from 147 U/L at 400 mg/kg to 164 U/L at 2400 mg/kg however this was not statistically significant and no mortality was recorded. In comparison to the negative control, treatment groups&rsquo; gross and histopathology of livers and kidneys had no significant difference. These results demonstrate that 80% ethanolic extract of E. sacleuxii roots has no severe effects in both acute and subacute study. However, sub-chronic and chronic toxicity studies are necessary for further validation. Key words: Erythrina sacleuxii, acute and sub-acute studies, phytochemical analysis.

Acute and Subacute Toxicity Studies of Cyperus Papyrus Ash on Wistar Albino Rats

Background: Acute and subacute toxicity screenings of medicinal plants are the fastest way to evaluate the toxicological profiles of medicinal plants. It helps to have a quick idea of the harm or safety potency of drugs. This study aims to conduct an acute and subacute oral toxicity study on the Cyperus Papyrus (CP) ash in Wistar albino rats. CP is traditionally used for managing painful spasms, eye diseases, ulcers, fever, diarrhea, and inflammations. Further study is required to reflect the safety profile of this plant ash. Methods: Acute oral toxicity study of the CP ash was evaluated by using a dose of 2000 mg/kg body weight on rats according to the Organization for Economic Co-operation and Development (OECD) guidelines using the Aot425 software. Subacute oral toxicity was carried out on 4 groups of 6 rats, at doses of 175 mg/kg, 550 mg/kg, 1750 mg/kg, and 1750 mg/kg. The recovery group (control group) received water according to OECD 407 guidelines for consecutive 28 days. At the end of the experiments (day 29), all rats were weighed and sacrificed except the rats of the recovery group which were continued without ash administration for extra 14 days and then weighed and sacrificed. Subsequently, the hematological parameters, plasma biochemical parameters, and histopathological examination were carried out. Results: Upon completion of the acute toxicity, no deaths or signs of acute oral toxicity were observed. Hematologic observations after oral subacute toxicity included a decrease in hematocrit percentage, mean corpuscular volume, and lymphocyte percentage, while there was a slight increase in the mean corpuscular hemoglobin concentration, however, it returned to the normal range in the recovery group. Biochemical tests showed a mild increase in the serum creatine and aspartate aminotransferase levels but they also returned to their normal range in the recovery group. No morphological changes in the kidney, liver, lung, spleen, heart or small intestine were observed after the histopathological examinations. Conclusion: Dried ash of CP is considered to be safe and non-toxic at acute exposure and as the lethal dose 50% value exceeds 2000 mg/kg body weight, it has a mild reversible effect on some hematological and biochemical parameters while no morphological changes of vital organ histopathology occur.

Safety Assessment of Lactiplantibacillus plantarum TWK10 Based on Whole-Genome Sequencing, Phenotypic, and Oral Toxicity Analysis.

Lactiplantibacillus plantarum TWK10 (TWK10), isolated from Taiwanese pickled cabbage, has been demonstrated to exert beneficial probiotic effects in both mice and humans. Here, we comprehensively assessed the safety of TWK10 using both in vivo and in vitro approaches, including whole-genome sequence analysis, an assessment of hemolytic activity, and performing an antimicrobial susceptibility test, the Ames bacterial reverse mutation assay, the chromosomal aberration test, a rodent peripheral blood micronucleus test, and the 28-day subacute oral toxicity assay. The results showed that there was no significant increase in the incidence of reverse mutations or chromosomal aberrations following exposure to TWK10. Moreover, no significant changes were detected either in the number of reticulocytes or the incidence of micronuclei in ICR mice, and no subacute toxicity was recorded in SD rats at the oral TWK10 dosage of 2000 mg/kg body weight/day repeated for 28 days. Additionally, TWK10 exhibited no hemolytic activity and was susceptible to all the antibiotics tested, except kanamycin. However, no antimicrobial resistance genes, virulence factors, or genes involved in biogenic amine synthesis were found in the genome of TWK10. Our findings demonstrated that TWK10 has high potential of being safe for human consumption as a probiotic.

GlucoMedix®, an extract of Stevia rebaudiana and Uncaria tomentosa, reduces hyperglycemia, hyperlipidemia, and hypertension in rat models without toxicity: a treatment for metabolic syndrome

BackgroundThe objective of this in vivo study is to evaluate in five rat models the pharmacologic effects and toxicity of a commercial hydro-alcoholic extract, GlucoMedix®, derived from Stevia rebaudiana and the pentacyclic chemotype of Uncaria Tomentosa (Willd.) DC, for use as a treatment for metabolic syndrome. The extract contains phytochemicals of Stevia (e.g., steviol glycosides) and Uncaria (e.g., pentacyclic oxindole alkaloids, but lacks tetracyclic oxindole alkaloids).MethodsThe pharmacologic assessments in three rat models include reductions in chemically induced hyperglycemia, hyperlipidemia (cholesterol and triglycerides), and hypertension, all of which are comorbidities of metabolic syndrome. Acute toxicity and 28-day subacute toxicity were assessed in rat models at doses higher than those used in the efficacy models.ResultsThe acute oral toxicity was evaluated in Holtzman rats and the extract did not produce acute toxic effects or lethality, with the LD50 > 5000 mg/kg (extract wet weight). Furthermore, subacute oral toxicity was evaluated in rats for 28 days at daily doses as high as 2000 mg/kg without toxicity or abnormal clinical chemistry or hematological effects. Daily oral doses of 250 - 1000 mg/kg were used to evaluate the treatment effects in hyperglycemic (alloxan-induced and glibenclamide-controlled), hyperlipidemic (cholesterol-induced and atorvastatin-controlled), and hypertensive (L-NAME-induced and enalapril-controlled) rat models. Alloxan-induced hyperglycemia was reduced in a dose-dependent manner within 28 days or less. Cholesterol-induced hyperlipidemic rats exhibited dose-dependent reductions in cholesterol and triglycerides at 21 days. Furthermore, GlucoMedix® produced a dose-dependent decrease in systolic and diastolic arterial blood pressure in L-NAME-induced hypertensive rats at 28 days.ConclusionsThe five in vivo rat models revealed that the all-natural phytotherapy GlucoMedix® is a safe and effective treatment for hyperglycemia, hyperlipidemia, and hypertension. This extract is expected to affect multiple comorbidities of metabolic syndrome, without any acute or subacute oral toxicity in humans. Although multiple prescription drugs are well known for the treatment of individual comorbidities of metabolic syndrome, no drug monotherapy concurrently treats all three comorbidities.

Oral Toxicity Studies, Histopathology, and Anti-Diabetic Activity of Polyherbal Extract in STZ Induced Diabetes in Rats

Background: Diabetes mellitus is a disease and endocrine disorder. It is a growing health problem in various countries. The prevalence of diabetes is rising worldwide; around 5.4% rise in patients with diabetes is expected by 2025. The World Health Organization (WHO) has reported that 173 million adults are suffering from diabetes mellitus in developing counties. In this research, estimation of glucose levels was performed in Wistar rats post-induction of diabetes by streptozotocin. Metformin has been used as a standard dose for treatment. This study demonstrated the acute oral toxicity and subacute oral toxicity of ethanolic extract of Saraca asoca leaves and Asparagus racemosus roots and evaluated the antidiabetic activity. Objective: This study aimed to evaluate the acute toxicity and sub-acute toxicity of the polyherbal ethanolic extract on vital organs. The visible changes in the organs of each group of Wistar rats were also determined in this study. Moreover, the hypoglycaemic action of the polyherbal extract of Saraca asoca and Asparagus racemosus was also explored. Methods: Wistar rats were divided into four groups for assessing acute oral toxicity. Groups were administered doses of 5,50, 300, and 2000mg/kg body weight of the polyherbal extract. Subacute oral toxicity studies were performed by administering a polyherbal extract of 250, 500, 1000mg/kg body weight. For evaluating the antidiabetic activity, animals were divided into 5 groups. In group 1 (normal control), saline was administered; in group 2, standard drug Metformin was administered; in group 3, no drug was given to rats with diabetes (Streptozotocin-Induced diabetic with 150mg/kg body weight), thus forming the diseased group. In group 4, ethanolic extract with 100mg/kg group was administered, and in group 5, ethanolic extract of 200mg/kg was administered. On the last day, the blood glucose levels and body weights of rats were examined. Histopathology studies were conducted by sacrificing the animals, isolating the organs, such as tissue pancreas, spleen, heart, lungs, liver, and kidney, and placing them on slides using a microscopic examination. Data were selected by searching the articles in the databases, such as NCBI, Web of Science and ScienceDirect, and PubMed from 1989 to 2020 by using keywords, such as “Antidiabetic,” “Saraca indica,” “Asparagus racemosus,” “ethanolic polyherbal extract,” “oral toxicity study,” “histopathology,” and “Streptozotocin.” Results: The polyherbal ethanolic extract of Saraca asoca and Asparagus racemosus at a dose of 100 mg/kg and 200 mg/kg showed better effects against streptozotocin-induced diabetic rats having 150 mg/kg body weight. Regarding acute toxicity and subacute oral toxicity studies, all the extracts showed significant values (P &lt;0.05) and were found to be safe and non-toxic. The body weight was also improved; no inflammation and erosion were seen on any organs of the Wistar rat in a histopathology analysis. Conclusions: The polyherbal ethanolic extract of Saraca asoca and Asparagus racemosus showed hypoglycaemic activity against STZ-induced diabetes in experimental Wistar rats. The results showed the beneficial effects of these ethanolic extracts, improving the body weight and changes in lipid metabolism and protecting the organs of Wistar rats, such as the liver, kidney, spleen, pancreas, lungs, and heart, against impairment of blood glucose. All organs were weighted, tissues from organs were isolated, then stained using eosin dye, and changes were observed by microscopy photos. No signs of inflammation and erosion were reported at the end of the study, indicating the healing action of the extract.

WITHDRAWN: Oral toxicity studies of Ethyl Acetate Polyherbal Extract Gel Formulation Obtained from Berries and Flaxseed (EAPEG-BF) in Wistar Rats

Background: Berries family and flaxseed have been used in ethnomedicinal practice for the treatment of various disorders and different ailments. Despite its beneficial uses, no studies on its polyherbal formulated gel combination toxicity potential have been reported. The present research study was designed to evaluate the acute oral toxicity (14 days) and sub-acute oral toxicity (repeated dose) study (28 days) of ethyl acetate polyherbal extract gel formulation obtained from berries and flaxseed according to OECD TG No. 423 and 407. Objective: Male Wistar Rats were taken for toxicological screening of acute and sub-acute oral toxicity as per OECD guidelines of ethyl acetate polyherbal extract gel formulation obtained from berries (Rubus idaeus, Vaccinium macrocarpon, Vaccinium angustifolium, Fragaria x ananassa) and flaxseed (Linum usitatissimum) at doses of 500 mg/kg, 1000 mg/kg, 2000 mg/kg and 5000 mg/kg b.w. p.o. Methods: The rats were orally treated and screened for various behavioural patterns and the clinical signs of toxicity after administration. In sub-acute oral toxicity screening, the highest dose of 5000 mg/kg was selected for repeated dose toxicity for 28 days. The data of the results showed that no toxic symptoms were observed up to the dose of 2000 mg/kg body weight. When testing is required a dose of 5000mg/kg, only one step. There was no toxic symptoms and mortality were observed up to the dose of 2000 mg/kg body weight during the 14 days of the observation. There is no visible change in the food intake and body weight when compared to control. Results: The toxicological study revealed that LD50 of the extract formulation gel was greater than 5000 mg/kg body weight p.o. Conclusion: In comparison to the control group, there were no significant changes in levels of biochemical parameters and haematological parameters. It was concluded from the data obtained in this research study that though LD50 is greater than 5000 mg/kg body weight; but there was no occurrence of observed adverse effects levels appeared in the liver, kidney, lipid profile and blood parameters.

Acute and Sub-acute Toxicity Studies of Ethanol and Chloroform Extracts of Solanum giganteum Jacq. Leaves

<p style="text-align: justify;"><strong>Objectives:</strong> To investigate the acute and subacute oral toxicity of ethanol and chloroform extracts <em>Solanum giganteum</em> Jacq (SG) leaves in a rodent for evaluating its safety profile. <strong>Methods: </strong><em>Solanum giganteum</em> Jacq leaves extract for the acute oral toxicity (300 to 5000 mg/kg) and subacute oral toxicity SG-Chloro (SGC 50, 200, 500 mg/kg) and SG-Alcohol (SGA 100, 500, 1000 mg/kg) studies were administered orally according to the guidelines 423 and 407 of OECD, respectively.<strong> Results:</strong> In the acute oral toxicity study, doses are administered by 300 mg/kg to 5000 mg/kg. SGA does not show toxicity up to 5000 mg/kg, SGC shows toxicity at a dose of 2000 mg/kg. In the subacute toxicity study, the daily oral treatment with extracts of SGA 100, 500, 1000 mg/kg, SGC 50, 200, 500 mg/kg of extracts for 28 days did not produce any death or hazard. Likewise, SGA 100, 500, 1000 mg/kg, SGC 50, 200 mg/kg of <em>Solanum giganteum</em> Jacq no significant changes were recorded in food intake, body weight gain, the biochemical and haematological parameter of control and treated rats. SGC 500 mg/ kg of significant changes were recorded in food intake and body weight gains were decreased compared to the control group. SGC 500 mg/kg of extract treaded group saw mild liver pathological findings characterized by abnormal hepatic configuration was observed with treated animals. <strong>Conclusion: </strong>The acute and subacute toxicity study<em> Solanum giganteum </em>Jacq alcohol extract is considered relatively safe on acute and subacute oral exposure. <em>Solanum giganteum</em> Jacq chloroform extract shows acute toxicity at dose 2000 mg/kg and in subacute toxicity study, higher dose 500 mg/kg. <p style="text-align: justify;"><strong>Key words:</strong> Solanum giganteum, Acute toxicity, Aubacute toxicity, OECD, SG-Chloro, SG-alcohol.