Pharmacological evaluation of the anxiolytic and sedative effects of Tilia americana L. var. mexicana in mice

The anxiolytic and sedative effects of Tilia americana L. var. mexicana (Schltdl.) Hardin inflorescence extracts and its acute toxicity were tested. Sodium pentobarbital (SP)-induced hypnosis potentiation (SPP), as well as ambulatory activity and anti-anxiety response in three different experimental models were evaluated with hexane and methanol extracts in mice. In order to determine the proper timing of assessments and to identify the most active extract, a 100mg/kg dosage of hexane, ethyl acetate and methanol crude extracts were tested on SPP after 15, 30 and 60min of the administration. Then a dose-response curve was made for the hexane (10-1000mg/kg) and methanol (10-300mg/kg) extracts in all experimental models. Both extracts produced a significant and dose-dependent lengthening in the time of SP, with the methanol extract being more potent than the hexane extract at 60min after administration. Moreover, a significant and dose-dependent attenuation in the anxiety-response in the plus-maze test and exploratory cylinder activity, but also a diminution in the ambulatory activity and in the head dipping response were observed resembling the response to diazepam. Acute toxicity was observed with less dose of methanol extract (LD(50)=375mg/kg) in comparison to the hexane extract (LD(50)>2900mg/kg). Results of the present study shows that Tilia americana var. mexicana possesses depressant activity on the CNS similar to the better-studied species of European Tilia and reinforces its use as anxiolytic and sedative in traditional medicine.     

Hydroalcohol extract and fractions of Stachys lavandulifolia Vahl: effects on spontaneous motor activity and elevated plus-maze behaviour

The present study aimed to investigate the anxiolytic effects of four fractions of Stachys lavandulifolia Vahl. The aerial parts of the plant were extracted with petroleum ether (PF), ethyl acetate (EF), butanol (BF) and water (AF) and tested for spontaneous motor activity and elevated plus-maze (EPM) behaviour in mice. The hydroalcohol extract (HE) and different fractions of S. lavandulifolia were administered intraperitoneally to male Syrian mice, at various doses, 30 min before the behavioural evaluation. The HE of S. lavandulifolia (at 50 mg/kg) increased the percentage of time spent (39%) and the percentage of arm entries in the open arms (53%). The HE (50 mg/kg), PF (25 and 50 mg/kg), EF (25 and 50 mg/kg) and AF (50 mg/kg) of S. lavandulifolia significantly increased the percentage of time spent and the percentage of arm entries in the open arms. The BF up to a dose of 50 mg/kg had no significant effects on any of the measured parameters in the EPM. The spontaneous locomotor activity was significantly decreased in animals injected with each plant fractions, compared with that of saline. The EF and AF showed the least and the most reduction in the activity, respectively. The anxiolytic effects of EF, PF and AF could be related to their content of flavonoids, phenylpropanoids or terpenoids.     

Anxiolytic effect of Rubus brasilensis in rats and mice

The aim of the present work was to investigate if infuse and ethanolic extracts (aqueous, butanolic and wax fractions) of Rubus brasiliensis Martius (Rosaceae) induce anxiolytic effect. The extracts were administered to male Wistar rats and Swiss mice per oral route, at 50, 100 and 150 mg/kg, 30 min before the behavioral evaluation in the elevated plus maze (EPM). Both infuse and wax ethanolic fraction at the dosage 150 mg/kg, vo, increased the number and the percentage of open arm entries of rats and mice. The aqueous and butanolic fractions, obtained from ethanolic extract, failed to induce anxiolytic effect. The treatment of mice with flumazenil (Ro 15–1788), 1.5, 2.0 and 2.5 mg/kg, i.p., 15-min before the administration of infuse or wax fraction, 150 mg/kg, vo, blocked the infuse or wax fraction-induced anxiolytic effect. The LD₅₀ for the wax fraction was 1000 mg/kg. In conclusion, the infuse and wax ethanolic fraction of R. brasiliensis present anxiolytic effect in rats and mice. In addition, it is suggested that the anxiolytic effect may be attributed at least to one liposoluble principle with low acute toxicity which may be acting as an agonist on GABA_A-benzodiazepine receptor complex.     

The anxiolytic effect of Sho-ju-sen, a Japanese herbal medicine, assessed by an elevated plus-maze test in mice

Sho-ju-sen (SK), a Japanese herbal medicine with a nourishing tonic action, is composed of a water extract of Kumazasa leaves (Sasa kurinensis Makino et Sibata) (SS), and ethanol extracts of Japanese red pine needles (Pinus densiflora Sieb. et Zucc) (PN) and Ginseng roots (Panax ginseng C. A. Meyer) (PX) in the ratio 8:1:1. In this study, an elevated plus-maze test in mice was carried out to assess whether SK had an anxiolytic effect. No significant change was observed in either the plus-maze or activity test following a single administration of SK (10 and 20 mL/kg p.o.). However, mice allowed a free intake of SK (10% solution) for 5 days and longer showed a significant prolongation of the time spent in the open arms (an anxiolytic effect), as long as that caused by the benzodiazepine anxiolytic diazepam (1 mg/kg p.o.). SK (1%, 3% and 30% solutions for 7 days) tended to develop the anxiolytic effect. Of the constituents of SK, SS (8% solution), but not PN (1% solution) or PX (1% solution), resulted in the anxiolytic effect. Except for a slight acceleration in the motor activity by PN (1% solution), no significant change in the motor activity was produced by any treatment with SK, SS or PX. The combined treatment of SK (10% solution) or SS (8% solution) with 1 mg/kg diazepam enhanced the anxiolytic effect. Flumazenil (0.1 mg/kg s.c.), a benzodiazepine receptor antagonist, alone did not change the time spent in the open arms. However, it completely reversed the anxiolytic effect of SK, SS and diazepam. The present results suggest that: (1) long-term treatment with SK develops an anxiolytic effect, (2) SS is the main constituent for the anxiolytic effect of SK, and (3) benzodiazepine receptors are involved in the anxiolytic effect of SK and SS.     

Flavonoids with antinociceptive and anti-inflammatory activities from the leaves of Tilia argentea (silver linden)

Silver linden, Tilia argentea Desf. ex DC (Tiliaceae) leaves are used in the treatment of common cold and bronchitis. In order to evaluate this information, antinociceptive and anti-inflammatory activities of the two main flavonoid glycosides: kaempferol-3,7-O-alpha-dirhamnoside and quercetin-3,7-O-alpha-dirhamnoside isolated from the leaves, were investigated. For the antinociceptive activity, p-benzoquinone-induced writhing test and for the anti-inflammatory activity, carrageenan-induced hind paw edema model in mice were used. Both compounds were shown to possess potent antinociceptive and anti-inflammatory activity at 50 mg/kg dose, per os, without inducing any apparent acute toxicity as well as gastric damage.     

HPLC/MS analysis and anxiolytic-like effect of quercetin and kaempferol flavonoids from Tilia americana var. mexicana

Aim of the study

Around the world, Tilia species have been used in traditional medicine for their properties as tranquilizer. Furthermore, Mexican species of Tilia have been grouped as Tilia americana var. mexicana, but their specific content in flavonoids is poorly described. In this study, inflorescences of Mexican Tilia were collected in three different regions of Mexico to compare their flavonoid content and anxiolytic-like response.

Materials and methods

Flavonoid content was analyzed by using an HPLC–MS technique. For anxiolytic-like response, Tilia inflorescences extracts (from 10 to 300 mg/kg, i.p.) were tested in experimental models (open-field, hole-board and plus-maze tests, as well as sodium pentobarbital-induced hypnosis) in mice.

Results

HPLC–MS analysis revealed specific peaks of flavonoid composition demonstrating some differences in these compounds in flowers and bracts depending on the region of collection. No differences in the neuropharmacological activity among these samples of Tilia were found. Moreover, their effects were associated with quercetin and kaempferol glycosides.

Conclusions

Dissimilarities in the flavonoid composition of Mexican Tilias might imply that these species must be re-classified in more than one species, not as a unique Tilia americana var. mexicana. Since quercetin and kaempferol aglycons demonstrated anxiolytic-like response and that no difference in the pharmacological evaluation was observed between these three Mexican Tilias, we suggest that this pharmacological effect of Tilia inflorescences involves these flavonoids occurrence independently of the kind of glycosides present in the samples reinforcing their use in traditional medicine in several regions of Mexico.     

基于成分敲除技术辨识灯心草抗焦虑的主要有效成分

目的 明确灯心草Juncus effusus抗焦虑作用的主要有效成分。方法 利用硅胶柱色谱技术,对灯心草干燥茎髓部80%乙醇提取物的醋酸乙酯超声萃取粗提物中量较高的菲类成分去氢厄弗酚、厄弗酚、去氢灯心草酚、灯心草酚进行敲除,制备相应的菲类目标成分、目标成分阴性样品,将菲类目标成分、等比例的目标成分阴性样品和全成分分别进行小鼠高架十字迷宫试验。结果 菲类目标成分与全成分能明显提高小鼠进入开放臂的时间和次数,作用强度无差异,而目标成分阴性样品未表现出抗焦虑活性。结论 菲类成分为灯心草抗焦虑作用的主要有效成分,灯心草中与菲类成分共存的其他成分对其抗焦虑作用基本无影响。     

Anxiolytic effects of Salvia reuterana Boiss. on the elevated plus-maze model of anxiety in mice

The anxiolytic and sedative effects of hydroalcoholic extract (HE) of Salvia reuterana Boiss. was evaluated in mice. The HE of Salvia reuterana (100 mg/kg), increased the percentage of time-spent and the percentage of arm entries in the open arms of the elevated plus-maze. Spontaneous locomotor activity count measured in 15 min of the test was significantly decreased in animals pretreated with diazepam and 100 mg/kg of Salvia reuterana extract. Results of the present study provide support for the traditional usage of Salvia reuterana as a sedative and anxiolytic medicinal plant.     

Hydroalcoholic extract and fractions of Davilla rugosa Poiret: effects on spontaneous motor activity and elevated plus-maze behavior

Davilla rugosa Poiret is commonly used in Brazilian folk medicine. The use as stimulant induced us to study the effects on motor activity and anxiety using an open-field and an elevated plus-maze, respectively. The hydroalcoholic extract of the stems (HE) was fractionated with chloroform (CF), chloroform/ethyl acetate (CAF), ethyl acetate (AF), ethyl acetate/ethanol (AEF), ethanol (EF) and ethanol/water (EWF). Rats were treated orally with HE (7.5, 15, 30 or 60 mg/kg) or fractions (15 mg/kg). In the open-field, HE (15 mg/kg), AEF, EF and EWF increased locomotion frequency and decreased immobility time; the contrary was observed with 30 and 60 mg/kg of HE. These doses also increased defecation. No effects were observed with 7.5 mg/kg of HE, CF, CAF or AF, except for an increase in defecation induced by AF. In the elevated plus-maze, total entries and number of entries into the open and closed arms and the time spent in the open arms and its percentage were increased only with 15 mg/kg of HE. The open-field results suggest that the drug increases motor activity (stimulant effect) and that the active components are in the three more polar fractions. An anxiolytic effect was observed only with the HE.     

Sedative and anxiolytic efficacy of Tilia americana var. mexicana inflorescences used traditionally by communities of State of Michoacan, Mexico

AIM OF THE STUDY: Increasing demand of herbal products acquired in stores and markets, as well as medicinal plants collected for personal consume are a known modern tendency. In this study, the ethnomedicinal use of Tilia americana var. mexicana inflorescences as sedative and anxiolytic is reinforced by examinating inflorescences used by communities of the State of Michoacan, Mexico. MATERIALS AND METHODS: Experimental mouse models were used to evaluate the sodium pentobarbital (SP)-induced hypnosis potentiation, ambulatory activity, as well as sedative and anti-anxiety responses via oral administration of the aqueous extracts (10, 30 and/or 100 and 300mg/kg). RESULTS: All samples tested produced a lengthening in the time of SP. Moreover, a significant attenuation in the anxiety-response in the plus-maze test and a diminution in both the head dipping response and ambulatory activity were observed resembling the response to diazepam (0.3mg/kg, i.p.). TLC profiles of the samples showed similar pattern of flavonoids; HPLC-DAD exhibited peaks identified as derived of quercetin and kaempferol that may be responsible for the plant activity. CONCLUSIONS: Our results demonstrate that inflorescences of stored specimens obtained from popular local markets show the same effectiveness with regard to sedative and anxiolytic-like actions than freshly collected samples. Since no toxicity was observed through this route of administration (up to 5000mg/kg); therefore, it suggests that this plant is secure when used as tranquilizer in folk medicine.     

Anxiolytic properties of Piper methysticum extract samples and fractions in the chick social-separation-stress procedure

Piper methysticum extract (Kava kava) possesses anxiolytic properties. However, it is unknown whether these effects are best predicted by total kavalactone content or by one or more of its primary kavalactone constituents. Using the chick social separation-stress procedure as an anxiolytic bioassay, P. methysticum samples containing 12.8-100.0% total kavalactones (Exp. 1) and fractions containing 1-6 kavalactones of varying concentrations (0.1-67.5%; Exps. 2-3) were screened for activity and compared against a 5.0 mg/kg dose of chlordiazepoxide (CDP; Exp. 3). Eight-day-old chicks received IP injections of either vehicle or test compounds 30 min before being placed in the presence of two conspecifics or in isolation for a 3 min observation period. Dependent measures were ventral recumbency latency (sedation), distress vocalizations, and a measure of stress-induced analgesia (in Exps. 1 and 2 only). P. methysticum extract samples attenuated distress vocalizations in a concentration-dependent manner. The P. methysticum fraction that contained the highest concentration of dihydrokavain attenuated distress vocalizations in a manner equivalent to that of CDP. The extract samples and fractions that possessed anxiolytic properties did not possess the sedative properties found in CDP. Collectively, these findings suggest that dihydrokavain may be necessary and sufficient in mediating the anxiolytic properties of P. methysticum extract.     

Anxiolytic and hypnotic effect of Crocus sativus aqueous extract and its constituents,crocin and safranal,in mice

Saffron stigma (Crocus sativus L.) is used for insomnia and anxiety in traditional medicine. In this study, the anxiolytic and hypnotic effects of saffron aqueous extract and its constituents, crocin and safranal, were studied in mice. Agents were administered intraperitoneally in mice before the experiments for the evaluation of hypnotic activity (induced by sodium pentobarbital, 30 mg/kg, i.p.), anxiolytic activity (elevated plus maze test), locomotor activity (open field test) and motor coordination (Rotarod test). The aqueous extract reduced the locomotor activity dose dependently. At low doses, saffron showed a significant increase in the time on the open arms of the maze. When using the Rotarod method, the aqueous extract showed considerable effect on motor coordination of the mice. In the hypnotic test, only a dose of 0.56 g/kg of saffron increased the total sleep. Crocin showed no anxiolytic, hypnotic or myorelaxation effects. Safranal, in higher doses, 0.15 and 0.35 mL/kg, showed anxiolytic effects. Safranal increased the total sleep time dose dependently. This constituent at lower doses (0.05 and 0.15 mL/kg) decreased some locomotion activity parameters. Safranal demonstrated no effects on motor coordination. The results showed that saffron aqueous extract and safranal have anxiolytic and hypnotic effects. Copyright © 2009 John Wiley & Sons, Ltd.     

Anxiolytic effect of seed of Ziziphus jujuba in mouse models of anxiety

The aim of the present study was to investigate the ethanolic extract of Semen Ziziphi jujuba (SZJE) induced anxiolytic effect. The SZJE was orally administered to male ICR mice, at 0.5, 1.0 and 2. 0 g/kg, 30 min before the behavioral evaluation in the black and white test (BWT) and elevated plus maze (EPM). The SZJE at the dosage 0.5-2.0 g/kg increased the first time entry, total changes and times spent in the white chamber of the BWT. The SZJE at the dosage 0.5-1.0 g/kg increased the percentage of time-spent and the percentage of arm entries in the open arms of the EPM and decreased the percentage of time-spent and the percentage of arm entries in the closed arms of the EPM. Furthermore, the SZJE at the dosage of 1. 0 g/kg prolonged the hexobarbital-induced sleeping time in mice and decreased the locomotor activity in rats. These results suggested that SZJE possessed anxiolytic effect at lower dose and sedative effect at higher dose.     

Possible anxiolytic effect of two extracts of Passiflora quadrangularis L. in experimental models

Several species of the genus Passiflora, known in Brazil as 'maracujá', have widespread use in folk medicine as sedatives and anxiolytics. The anxiolytic activities of aqueous and hydroalcohol extracts of Passiflora quadrangularis leaves were evaluated using the elevated plus-maze, open field and holeboard tests. The hydroalcohol extract presented results suggestive of anxiolytic activity in dosages around 100, 250 and 500 mg/kg, as expressed by elevation of the time spent on the open arms in the plus-maze; a decrease of freezing and an increase of deambulation and rearing in the open field test. The hydroalcohol extract showed results similar to diazepam on the holeboard. No positive results were found for the aqueous extract.     

Postnatal development and reproductive performance of F1 progeny exposed in utero to an ayurvedic contraceptive: Pippaliyadi yoga

The purpose of this study was to characterize the putative anxiolytic-like activity of an ethanolic extract prepared from the leaves of Apocynum venetum (AV) using the elevated plus maze (EPM) in mice. Male C75BL/6 mice were either treated orally with the AV extract or the positive controls diazepam and buspirone, respectively, 1 h before behavioral evaluation in the EPM. A single treatment of AV extract markedly increased the percentage time spent on and the number of entries into the open arms of the EPM in doses of 30 and 125 mg/kg p.o., respectively. This effect was comparable to that of the benzodiazepine diazepam (1.5 mg/kg p.o.) and the 5-HT_1A agonist buspirone (10 mg/kg p.o.). The effects of AV in 125 mg/kg were effectively antagonized by the benzodiazepine antagonist flumazenil (3 mg/kg i.p.). However, the effects of AV extract could only partially be blocked by the unspecific 5-HT_1A receptor antagonist WAY-100635 (0.5 mg/kg i.p.). Neither diazepam and buspirone nor the AV extract produced any overt behavioral change or motor dysfunction in the open field test. These results indicate that AV extract is an effective anxiolytic agent, and suggest that the anxiolytic-like activities of this plant are mainly mediated via the GABAergic system.     

Coriandrum sativum: evaluation of its anxiolytic effect in the elevated plus-maze

The clinical applications of benzodiazepines as anxiolytics are limited by their unwanted side effects. Therefore, the development of new pharmacological agents is well justified. Among medicinal plants, Coriandrum sativum L. has been recommended for relief of anxiety and insomnia in Iranian folk medicine. Nevertheless, no pharmacological studies have thus far evaluated its effects on central nervous system. Therefore, the aim of this study was to examine if the aqueous extract of Coriandrum sativum seed has anxiolytic effect in mice. Additionally, its effect on spontaneous activity and neuromuscular coordination were evaluated. The anxiolytic effect of aqueous extract (10, 25, 50, 100 mg/kg, i.p.) was examined in male albino mice using elevated plus-maze as an animal model of anxiety. The effects of the extract on spontaneous activity and neuromuscular coordination were assessed using Animex Activity Meter and rotarod, respectively. In the elevated plus-maze, aqueous extract at 100 mg/kg showed an anxiolytic effect by increasing the time spent on open arms and the percentage of open arm entries, compared to control group. Aqueous extract at 50, 100 and 500 mg/kg significantly reduced spontaneous activity and neuromuscular coordination, compared to control group. These results suggest that the aqueous extract of Coriandrum sativum seed has anxiolytic effect and may have potential sedative and muscle relaxant effects.     

Comparison of antimalarial activity of the alkaloidal fraction of Hydrangea macrophylla var. Otaksa leaves with the hot-water extract in ICR mice infected with Plasmodium yoelii 17 XL

The antimalarial activity of the fractions isolated from the leaves of Hydrangea macrophylla Seringe var. Otaksa Makino was evaluated against Plasmodium yoelii 17 XL in mice. Four different fractions were prepared in the usual manner to obtain an alkaloid fraction. All mice treated with the fraction containing febrifugine and isofebrifugine mixture at 1 mg/kg twice a day for 5 consecutive days survived during the experiment, and the change of mean parasitemia level showed almost the same pattern as that from mice treated with the hot-water extract of the same plant leaves. Activity of this fraction, however, was markedly reduced compared with the hot-water extract. Furthermore, no antimalarial activity was shown in the hotwater extract from H. macrophylla var. Otaksa roots or Dichroa febrifuga Lour. leaves.     

Application of the elevated plus-maze test in mice for evaluation of the content of honokiol in water extracts of magnolia.

In our previous study using an improved elevated plus-maze in mice, the oriental herbal medicine Saiboku-to prolonged the time spent in open arms, showing an anxiolytic effect, and the effect was mainly caused by honokiol derived from magnolia. This study was carried out to compare the anxiolytic potentials of honokiol and water extracts of three magnolia samples; two being Kara-koboku (Magnolia officinalis) (KA: from Zhejiang-sheng, China; honokiol 0.25% and magnolol 1.16%, and KB: from Sichuan-sheng, China; honokiol 1.72% and magnolol 1.71%), and one being Wa-koboku (Magnolia obovata) (WA: from Iwate-ken, Japan; honokiol 0.32% and magnolol 0.81%). Seven daily treatments with 0.1-1 mg/kg honokiol, but not 0.2 and 1 mg/kg magnolol, revealed an anxiolytic effect with the peak potential at 0. 2 mg/kg. The anxiolytic potentials of 40 and 80 mg/kg KA, which contained the highest amount of magnolol, were almost equivalent to those of 0.1 and 0.2 mg/kg honokiol, respectively. KB, at 11.6 mg/kg, and 62.5 mg/kg WA resulted in almost the same anxiolytic potential as that of 0.2 mg/kg honokiol. No significant change in the ambulatory activity was produced by any drug treatment. These results suggest that honokiol is the chemical responsible for the anxiolytic effect of the water extract of magnolia and that the other chemicals including magnolol in magnolia scarcely influence the effect of honokiol. It is also considered that the elevated plus-maze test is applicable for evaluation of the content of honokiol in magnolia.     

Neurosedative and muscle-relaxant activities of ethyl acetate extract of Baphia nitida AFZEL

The sedative, anxiolytic and muscle-relaxant effects of the ethyl acetate leaf extract of Baphia nitida (BN) was investigated in intact mice, using the hole-board head-dip test for exploratory behavioural effect, elevated plus maze (EPM) and Y-maze (YM) models of anxiety; chimney, inclined screen, traction and climbing tests for muscle-relaxant effects. In each of these tests, BN (100-400mg/kg, p.o.), diazepam (1mg/kg, i.p.) or distilled water (10ml/kg, p.o.) was administered, 30 or 60min before performing the tests in mice. For exploratory behavioural test, number of head-dip within 15min was counted. For EPM and YM tests, the cumulative time spent in open and closed arms was recorded within 5min. In the muscle-relaxant tests, mice were subjected to modified models such as chimney, inclined screen, traction and climbing tests. BN produced a significant (P<0.05) dose-related decrease in exploratory behaviour in the head-dip test and prolongation of cumulative time spent in open arms of both EPM and YM. BN did not show any significant effect in the chimney and traction tests, but produced significant, dose-dependent muscle relaxation in the inclined screen and climbing tests. Furthermore, BN (200-1200microg/ml) non-competitively shifted the curves of acetylcholine contractions of the toad Rectus abdominis muscle to the right. Oral doses of BN (0.1-20g/kg) did not produce mortality, but the LD(50) when given intraperitoneally, was 645.65mg/kg. Results suggest that the leaf extract of Baphia nitida has sedative, anxiolytic and skeletal muscle-relaxant effects and support its neurosedative use in traditional African medicine.     

CNS activity of the methanol extract of Mallotus peltatus (Geist) Muell Arg. leaf: an ethnomedicine of Onge

The aim of the present study was to investigate several neuropharmacological effects of the methanol extract and different fractions of Mallotus peltatus (Geist) Muell Arg. var acuminatus (Euphorbiaceae) leaves in Wistar albino rats and Swiss albino mice. General behavior, exploratory behavior, muscle relaxant activity and phenobarbitone sodium-induced sleeping time were studied. The results revealed that the crude extract at 200-300 mg kg(-1) p.o. and its fractions A and B at 50 mg kg(-1) caused a significant reduction in spontaneous activity (general behavioral profile), remarkable decrease in exploratory behavioral pattern (Y-maze and head dip tests), a reduction in muscle relaxant activity (rotarod, 30 degrees inclined screen and traction tests), and also significantly potentiated phenobarbitone sodium-induced sleeping time. The phytochemical study of crude leaf extract revealed the presence of tannin, triterpenoid, flavonoid, sterol, alkaloid and reducing sugar. Further fractionation and purification yielded two major fractions A (ursolic acid) and B (beta-sitosterol) with some fatty acids as the major compounds. The psychopharmacological activity of the crude leaf extracts appeared to be either due to fraction A (50 mg kg(-1)) or a combination of fractions A and B (50 mg kg(-1)) along with some fatty acids present in the n-butanol part of methanol extract of M. peltatus leaf (MEMPL).