Different processing methods change the oral toxicity induced by Sophora alopecuroides seeds and the contents of five main toxic alkaloids from the ethanol extracts determined by a validated UHPLC-MS/MS assay

Abstract This study investigated the influence of different processing methods on the oral toxicity of Sophora alopecuroides L., Fabaceae, seeds in mice and on the contents of five known toxic-effective quinolizidine alkaloids from the ethanol extracts quantified by ultra-performance liquid chromatography coupled to tandem quadrupole mass spectrometry. It provides an evidence to elucidate the possible reasons why vinegar-processing and parching methods significantly decrease the acute oral toxicity induced by S. alopecuroides and why wine-processing method increases it instead (demonstrated by measurement of LD50 and histopathological analysis). The analytical performance for the determination of the five analytes was evaluated by linearity, stability, repeatability, precision and accuracy, and recovery test. The lowest limit of quantification was determined to be 5 ng/ml for each substance and the precision and accuracy at lowest limit of quantification were below 20%. Cytisine, the most toxic alkaloid among the five alkaloids, declined 11.26, 3.98, and 2.73 folds after being vinegar-processed and fried in a ceramic or iron pan, respectively and had a very close correlation with the toxicity of S. alopecuroides seeds (r = 0.8589). Other matrine-type alkaloids with lower toxicity including matrine, sophcarpine, and sophoridine decreased after being wine-processed and fried in a ceramic pan, but increased 4.44, 7.20, and 7.23 folds when being processed by vinegar. Oxymatrine declined in all groups. It, therefore, reveals that vinegar-processing method reduces the oral toxicity of S. alopecuroides mainly due to a sharp decrease of cytisine, thus improves its clinical safety.

Preclinical safety assessment of glycosides based standardized fenugreek seeds extract: Acute, subchronic toxicity and mutagenicity studies.

Objective: To evaluate acute oral toxicity (AOT), subchronic toxicity, and mutagenic potential of glycosides based standardized fenugreek (Trigonella foenum graecum L.) seeds extract (SFSE-G). Materials and Methods: The AOT, subchronic (90-day repeated dose) toxicity and mutagenicity (reverse mutation test) of oral administration of SFSE-G were evaluated using Sprague-Dawley (SD) rats as per OECD guideline no. 423, No. 408 and 471 respectively. Results: The SFSE-G did not show mortality or treatment-related adverse signs during acute (limit dose of 2000 mg/kg) and subchronic (90-days repeated dose of 250, 500 and 1000 mg/kgwith 28 days of recovery period) administration. The SFSE-G showed oral median lethal dose (LD50) more than 2000 mg/kg during AOT study. The no-observed adverse effect level (NOAEL) of SFSE-G was 1000 mg/kg in male rats and 500 mg/kg in female rats during subchronic toxicity study. Furthermore, SFSE-G did not show mutagenic potential in vitro. Conclusions: SFSE-G was found safe for acute and subchronic (90 days repeated dose) administration in rats with no mutagenic potential.

Acute oral toxicity and genotoxicity of Rosa laxa Retz. var. mollis Yu et Ku cynarrhodion extracts / 国际药学研究杂志

Objective To study the acute oral toxicity and genotoxicity of Rosa laxa Retz. var. mollis Yu et Ku cynarrhodion extracts. Methods The acute oral toxicity in mice and rats (maximal tolerance dose, MTD) were observed for 14 days after administration. Genetictoxicity was evaluated on Salmonella typhimurium reverse mutation assay (Ames test), in vivo mammalian erythrocyte micronucleus assay, sperm malformation assay in mice and single cell gel electrophoresis (Comet assay). Results In acute oral toxicity study, the behavior, clinical signs and body changes were normal in all experimental animals. LD50 > 15.0 g/kg (rats) and 20.0 g/kg (mice). In the test dose range, results of the four genotoxicity tests were negative. Conclusions In accordance with internationally recognized animal ethics and technical specifications for the experiments using SPF animals in our lab, the acute oral toxicity of R. laxa Retz. var. mollis Yu et Ku cynarrhodion extracts is classified as nontoxic, and genotoxicity is not observed under the test condition.

Isolation of the intracellular and extracellular polysaccharides of Ganoderma neojaponicum (Imazeki) and characterization of their immunomodulatory properties

Background The role of polysaccharides isolated from the Ganoderma species of fungi in innate immunity has recently become a topic of research. Although some work has been conducted concerning Ganoderma lucidum, the characteristics of polysaccharides isolated from Ganoderma neojaponicum (Imazeki) as immunomodulatory agents are largely unknown. The aims for this study were to isolate and characterize the intracellular polysaccharides (IPSs) and extracellular polysaccharides (EPSs) of G. neojaponicum from STR reactor. Results The production of EPS and IPS was optimized on day 4 of the cultivation time in 2 L STR reactor based on the amount of biomass yield, total carbohydrate, β-glucan and a-glucan content. Further analysis, both the EPSs and IPSs showed the enhancement on proliferation and increment of phagocytosis activities of macrophage (RAW264.7) cell lines. Using an oral toxicity test, we also observed that 2000 mg/kg body weight/day dosage of dried G. neojaponicum mycelium does not cause any significant toxic effects on Sprague-Dawley rats in 14 d of administration. Conclusion The findings of this study indicate that the IPSs and EPSs of G. neojaponicum have the potential to be used as immunomodulating agents to stimulate the innate immune system for fighting infectious diseases. The polysaccharides from G. neojaponicum have to be further commercially explored as an alternative for medicinal Ganoderma variety of G. lucidum production.

Acute oral toxicity of Celtis iguanaea (Jacq. ) Sargent leaf extract (Ulmaceae) in rats and mice / Toxicidade oral aguda do extrato de folhas de Celtis iguanaea (Jacq. ) Sargent em ratos e camundongos

RESUMO Celtis iguanaea (Jacq.) Sargent is popularly used to treat urinary infections, kidneys, breast, body aches, rheumatism, asthma, cramps, poor digestion and as a diuretic medicine. This study aims to determine the acute toxicity of the aqueous leaf extract of Celtis iguanaea (Jacq.) Sargent in rodents. After the collection processes, identification, drying and grinding, the lyophilized powder of the leaves produced, by infusion, the aqueous extract and it was dissolved in saline 0.9%. The administration was made by gavage at a dose of 2000 mg kg-1to rats and mice of both genders. The oral toxicity was determined according to the OECD 423 guide. Signs of toxicity were observed for 15 days and classified from 0 to 4 respectively as missing, rare, mild, moderate and severe. The weight of the animals and the physiological parameters such as food intake and excrements production were observed. All animal tissue samples were collected for histological analysis. The extract was included in Type 5 (substance with LD50 higher than 2000 mg kg-1 and less than 5000 mg kg-1), being considered of low toxicity, but the histopathologycal findings suggested nephrotoxicity and cardiotoxicity. The absolute weight of the kidneys and the heart of the male rats and mice increased, but there was no significant raise in the relative weight of the animals’ organs.

RESUMO Celtis iguanaea (Jacq.) Sargent é uma planta usada popularmente para tratar infecções do trato urinário, rim, mama, dores no corpo, reumatismo, asma, cólicas, má digestão e também é usada como diurético. Este trabalho objetivou determinar a toxicidade aguda do extrato aquoso de folhas de Celtis iguanaea (Jacq.) Sargent em roedores. Após os processos de coleta, identificação, secagem e moagem, o pó liofilizado das folhas da planta foi utilizado para produzir o seu extrato aquoso por infusão e então dissolvido em solução salina a 0.9 %. A administração foi feita por gavagem na dose de 2000 mg kg-1 em ratos e camundongos de ambos os sexos. A toxicidade oral foi determinada de acordo com o guia 423 da OECD. Sinais de toxicidade foram observados por 15 dias e tabulados de 0 a 4, respectivamente, como ausentes, raros, leves, moderados e graves. Foi acompanhado o peso dos animais e parâmetros fisiológicos tais como alimentação e excreções. Amostras do tecido de todo o animal foram coletadas para análise histológica. A toxicidade encontrada para o extrato foi incluída na classe 5 (substâncias com DL50 superior a 2000 mg kg-1 e menor que 5000 mg kg-1) sendo considerada baixa, porém, as observações histopatológicas sugerem nefrotoxicidade e cardiotoxicidade. O peso absoluto dos rins e coração de ratos e camundongos machos aumentou, porém, não houve aumento significativo no peso relativo dos órgãos dos animais.

Assessment of acute toxicity of the ethanolic extract of Lychnophora pinaster (Brazilian arnica)

Species of the Lychnophora genus are plants native to Brazil, popularly known as "Brazilian arnica" and used in folk medicine as alcoholic and hydro-alcoholic preparations for the treatment of bruises, inflammation, pain, rheumatism and insect bites. The present study aimed to evaluate the safety of the use of Lychnophora pinaster Mart., Asteraceae. Acute toxicity of the crude ethanolic extract was evaluated by administration of the extract by oral route to male and female Swiss mice. A single extract dose of 125, 250 or 500 mg/kg was administered and the effects on spontaneous locomotor activity, exploratory behavior, muscle strength, body weight, food and water consumption, relative organ weight, histology, as well as hematological and biochemical parameters were evaluated. The three doses administered to the animals did not cause muscle tone alterations, but doses of 250 and 500 mg/kg induced a significant inhibition of the spontaneous locomotor activity and exploratory behavior of the animals in open-field test. There was no alteration to hematological parameters and consumption of water and food, body weight variation and organs relative weight. Changes were observed in AST and ALT during assessment of biochemical parameters. The histopathological evaluation showed that the extract provoked cellular alterations, such as vacuolar degeneration and inflammation in kidneys and liver at all doses. Liver morphometric analyses of male and female mice showed that the extract did not have dose-dependent effects. Although females showed a significant increase in inflammatory cells, the effect was not dose-dependent.

Acute oral toxicity and anti-inflammatory activity of hydroalcoholic extract from Lampaya medicinalis Phil in rats

BACKGROUND: Algesia and inflammation are related with several pathological conditions. It is known that many drugs available for the treatment of these problems cause unwanted side effects. This study was aimed at evaluating acute toxicity and anti-inflammatory activity of Lampaya medicinalis Phil. (Verbenaceae) widely used in the folk medicine of Northern Chile against rheumatism, arthritis and body joints pain. RESULTS: Oral administration of hydroalcoholic extract (HAE) at the highest dose of 3000 mg/ Kg body weight resulted in no mortalities or evidence of significant behavioral changes. Histological examination revealed normal architecture and no significant adverse effects were observed on the liver, kidney, heart, lung or ovaries and testicles. The results suggest that the oral administration of hydroalcoholic extract (HAE) from Lampaya medicinalis did not produce any toxic effect in rats. Hydroalcoholic extract (HAE) significantly inhibited the carrageenan-induced rat paw edema in dose - response relationship, at test doses of 37.5, 75, 150 and 300 mg/Kg body weight. Maximum inhibition (61.98 ± 2.69%) was noted at 300 mg/Kg after 2 h of drug treatment carrageenan induced paw edema, whereas indomethacin produced 47.90 ± 1.16% of inhibition. The inhibitory values of edema at 3 h postcarrageenan were 31.04±0.75%, 40.51 ± 2.36%, 48.97 ± 1.14% and 56.87 ± 0.41% for 37.5, 75, 150, and 300 mg/kg of extract respectively. Indomethacin (10 mg/Kg) gave a percentage inhibition of 49.44 ± 1.44. HAE (300 and 150 mg/kg) induced an anti-inflammatory effect greater than (or comparable) with the effect of indomethacin from 2nd to 4th hours of the experiment. CONCLUSIONS: Our results reveal for first time that compounds contained in the hydroalcoholic extract ofLampaya medicinalis Phil exert anti-inflammatory effect and the oral administration is safe and non toxic up to dose level 3000 mg/kg body weight. The anti-inflammatory activity may be associated with the presence of flavonoids. These findings also justify the traditional use of the plant for treating pain.

Anti-Inflammatory Potential of Ethanolic Extracts from Aerial Parts of Ipomoea Pes-Caprae (L.) R.Br Using Cotton Pellet Induced Granuloma Model.

Ipomoea Pes-caprae (L.) R.Br (IP) is a valuable medicinal plant, distributed in the tropics and subtropics regions and used in folk and tribal medicines. Traditionally IP is used in inflammatory conditions such as arthritis and also used to treat pain, ulcer, cancer and wounds. The acute anti-inflammatory activity of IP has been previously reported. The present study aims to discover the anti-inflammatory effect of ethanolic extracts from aerials parts of IP by sub-acute anti-inflammatory model. Completely dried leaves and stems of I.pes-caprae were extracted using ethanol by hot percolation method. The EELIP & EESIP (Ethanolic extract of Leaves & Stems of IP) thus obtained were subjected to preliminary phytochemical analysis and revealed the presence of alkaloids, carbohydrates, glycosides, flavonoids, tannins, sterols and terpenoids both in leaf and stem extracts. The LD50 of both EELIP & EESIP were found to be >2000 mg/kg by acute oral toxicity study. Both EELIP & EESIP exhibited significant anti-inflammatory activities in dose dependent manner.

In vitro antimicrobial potential of Terminalia chebula fruit extracts against multidrug-resistant uropathogens

Objective: Terminalia chebula Retz. (combretaceae) is called the “King of Medicine” in Tibet and is always listed at the top of the list of “Ayurvedic Materia Medica” because of its extraordinary power of healing. The present study was carried out to evaluate the possible in vitro antibacterial potential of different solvent extracts of T. chebula fruit against multidrug-resistant uropathogens. Methods: A total of 52 multidrug-resistant uropathogenic bacteria were used in this study. Successive extractions of T. chebula fruits were performed with solvents of different polarities. Agar well diffusion and microbroth dilution assay methods were used for antibacterial susceptibility testing. Kill-kinetics study was done to know the rate and extent of bacterial killing. Qualitative phytochemical screening was done to know the major phytoconstituents present in the plant material. Acute oral toxicity study in mice was performed to evaluate the toxic potential of the plant material, if any. Results:The ethanol extract of T. chebula fruits demonstrated a strong antimicrobial activity against all the test isolates and found to be most effective over others. Kill-kinetics study showed dose and time dependent antibacterial activity of ethanol extract. Phytochemical analysis revealed the presence of high concentration of phenolics and low concentration of flavonoids and terpenoids. In acute oral toxicity study, no gross behavioral changes were observed in mice at recommended dosage level and 24 h LD50 of ethanol extract was found to be >4 g/kg, p.o. in mice. Conclusions: The results provide justification for the use of Terminalia chebula fruit in folk medicine to treat various infectious diseases and could be useful for the development of alternative/ complementary medicine for multidrug-resistant uropathogens.

Pharmacognostical, antioxidant and antiulcer screening of Cyclea peltata roots

Cyclea peltata Hook. f. & Thomson, belongs to the family Menispermaceae commonly known as "paatha", is a climbing shrub found throughout South and East India. In Indian traditional medicine the tuberous roots are also used to treat jaundice, stomachache, fever and asthma. This study deals with the microscopic study of leafs and roots of C. peltata, along with the physico-chemical, preliminary phytochemical analyses and antioxidant properties. Acute oral toxicity study was conducted as per OECD-423 guidelines and the extract was found to be devoid of any conspicuous acute toxicity in extract treated animals and no mortality upto 2 g/kg by oral route. Hydroalcoholic extract of C. peltata in a dose dependant manner (125 and 250 mg/kg. p.o.) showed significant gastric protection against the ethanol-induced gastric ulcer model in rats.

Pharmacognostical standardization and toxicity profile of the methanolic leaf extract of Plectranthus amboinicus (Lour) Spreng.

The present investigation was intended to evaluate the toxicity of the methanolic leaf extract of a traditionally used plant Plectranthus amboinicus (Lour) Spreng. Plant material was analysed for various pharmacognostical parameters as per WHO guidelines procedure i.e., foreign matter, microscopical sections, loss on drying, water and alcoholic extractive values, Total ash, acid soluble ash, heavy metals, phytochemical analysis and toxicity studies. Acute & Sub acute toxicity of the methanolic extract was evaluated in albino mice (Female) after ingestions of the extract during one day (Acute model) and during 28days (sub acute model).The studies on sub acute toxicity reveals that no mortalities or evidence of adverse effects have been observed in Albino mice following acute oral administration at the highest dose of 2000mg/kg crude extracts of Plectranthus amboinicus (Lour) Spreng. Similarly, in sub-acute toxicity study methanolic extract 200,400 mg/kg body wt of Plectranthus amboinicus did not cause any changes in hematological and biochemical parameters. Studies on histopathological examination of vital organs showed normal architecture suggesting no morphological disturbances. Plectranthus amboinicus (Lour) Spreng can be considered as safe as it did not cause either any lethality or adverse changes in the general behavior in mice.

Evaluation of the toxicity and antiulcerogenic activity of the ethanol extract of Maytenus obtusifolia Mart. leaves / Avaliação da toxicidade e atividade antiulcerogênica do extrato etanólico das folhas de Maytenus obtusifolia Mart

Maytenus obtusifolia é utilizada na medicina popular para o tratamento de úlceras graves, inflamações gerais e câncer. Apesar da importância etnofarmacológica desta espécie, nenhum estudo foi realizado para avaliar a sua toxicidade e atividade antiulcerogênica. Neste estudo, nós avaliamos a toxicidade e propriedade antiulcera do extrato etanólico das folhas de Maytenus obtusifolia (MO-EtOH). O MO-EtOH (10-1000 µg/mL) mostrou baixa toxicidade para as larvas de A. salina com CL50 maior que 1000 µg/mL. O MO-EtOH (2000 mg/kg, p.o.) nγo alterou o peso corporal e peso dos órgãos dos camundongos, mas foi observado um aumento no consumo de água dos machos e uma diminuição do consumo alimentar das fêmeas. Durante o estudo não foram observadas mortes e nem alterações macroscópicas nos órgãos dos camundongos. MO-EtOH (62,5, 125, 250 e 500 mg/kg) e lansoprazol (30 mg/kg) reduziram significativamente o índice ulcerativo para 65,58 ± 8,74, 43,00 ± 9,53, 15,50 ± 7,56, 54,75 ± 8,88 e 36,13 ± 9,55, respectivamente, em comparação com salina 82,13 ± 12,48. Em conclusão, o MO-EtOH apresentou baixa toxicidade e atividade antiulcerogênica, o que confirma o uso popular de M. obtusifolia.

Maytenus obtusifolia is used in folk medicine for the treatment of serious ulcers, general inflammations and cancer. Despite of the ethnopharmacological importance of this species, no study was conducted to evaluate its toxicity and antiulcerogenic activity. In this study, we evaluated the toxicity and antiulcerogenic property of the ethanol extract of the leaves of Maytenus obtusifolia (MO-EtOH). The MO-EtOH (10-1000 µg/mL) showed low toxicity for larvae of A. salina with LC50 higher than 1000 µg/mL. The MO-EtOH (2000 mg/kg, p.o.) did not change the body and organs weight of the mice, but it was observed an increase in the water consumption of males and a decrease in the food consumption of females. During the study no deaths and no macroscopic changes in the organs were observed in the mice. MO-EtOH (62.5, 125, 250 and 500 mg/kg) and lansoprazole (30 mg/kg) significantly reduced the ulcerative index for 65.58 ± 8.74, 43.00 ± 9.53, 15.50 ± 7.56, 54.75 ± 8.88 and 36.13 ± 9.55, respectively, in comparison with saline 82.13 ± 12.48. In conclusion, the MO-EtOH showed low toxicity and antiulcerogenic activity, confirming the popular use of M. obtusifolia.

Acute Toxicity of Administered Bisphenol A Di Glycidyl Ether in Male Sprague Dawley Rats / 대한산업의학회지

OBJECTIVES: Bisphenol A Di Glycidyl Ether (BADGE) is the major component in commercial liquid epoxy resins, which are manufactured by co-reacting bisphenol A with epichlorohydrin. The authors investigated the acute toxicity of BADGE. METHODS: BADGE was administered by a gavage to 8 week old SPF Sprague Dawley rats in a single dose of 0 (negative control), 0.37 (Diethylstilbesterol, DES), 1000, 2000, 4000, and 8000 mg/kg/day of BADGE. Each treatment group contained 7 rats. The general status and weight of the rats were observed for 14 days. The rats were anesthetized by ether at 14 days, and the changes in morphology, organ weight, sperm count and motility, and hormone level were measured. RESULTS: All the rats treated with BADGE had diarrhea on the 1st day. The rats administered BADGE at 1000, 2000, and 4000 mg/kg/day showed a soiled perineal region and soft stools with diarrhea until the 3rd day. The 8000 mg/kg/day BADGE rats had diarrhea for two days followed by emaciation, soiled fur, a soiled perineal region, staining around the mouth and were moribund for three to eight days. No weight gain was observed after the 1st day in the 2000, 4000, and 8000 mg/kg/day BADGE rats and after the 7th day in all the treatment groups compared with the control groups. Some treatment groups were observed to have a decrease in the weight of the heart (BADGE 1000, 2000 and 4000 mg/kg/day), liver (BADGE 1000, 2000, 4000 and 8000 mg/kg/day) and prostate (BADGE 4000 mg/kg/day) compared with control group. The weight of the liver was significantly lower in all treatment groups compared with the control group. The relative weight of the liver (BADGE 1000 and 4000 mg/kg/day) was significant lower than the control. No pathological changes were observed in the brain, liver, thyroid, heart, spleen, kidney, lung and prostate. The number of spermatid in the seminiferous tubule in the testes was lower in all treatment groups than the control. The sperm motility tended to decrease with increasing concentration but the sperm count was similar in all treatment groups. The plasma Estrogen and testosterone level were similar in the control and treatment groups. CONCLUSIONS: These results suggest that BADGE induces general, hepatic and reproductive toxicity at 1000 mg/kg/day.