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Online First                                                                                                                      Indian J Pharm Drug Studies | 1  

Original Article 

EVALUATION OF ANXIOLYTIC ACTIVITY OF ETHANOLIC EXTRACT 

OF Phyllanthus niruri LEAVES IN MICE 

Spandana Savvaser1, Rakshitha Poojary1, Soubiya Mahnoor Habibullah1, Ashoka Shenoy M2 

From, 1Student, 2Associate Professor, Department of Pharmacology, Srinivas College of Pharmacy, Mangalore, India 

ABSTRACT 

Anxiety has become one of the most prevalent and widely experienced mental illnesses, affecting 7-30% of the world’s population. It 

is characterized as a distressing psychological state of mind, which is represented by internal feeling of uneasiness, stress and agony. 

Various theories, including psychodynamic, psychoanalytic, and genetic perspectives, have been proposed to explain the etiology of 

anxiety disorders. Certain drugs used for treatment of anxiety have side effects and limitation such as sedation, central complexity, 

and habituation and during withdrawal of drug. Researchers are actively exploring natural remedies, especially medicinal plants, to 

address anxiety disorders and minimize potential side effects associated with conventional drugs. Hence in the present study we 

investigate the anxiolytic activity of the plant Phyllanthus niruri leaves in mice using various experimental models such as Elevated 

Plus Maze and Light And Dark Chamber. In the Elevated Plus Maze (EPM) model, parameters measured included the number of 

entries in open and closed arms, along with the time spent in each. In the Light and Dark Chamber, measurements encompassed the 

number of crossings between light and dark sides recorded for 10 minutes, along with the time spent in each side. The administration 

of the different doses of the plant extract produced significant difference in the locomotor activity of the mice. 

Key words: Anxiety, Phyllanthus niruri leaves, EPM, Light and Dark Chamber. 

nxiety is defined as a cognitive emotional response 

characterized by anatomical activation, typically of 

the autonomic nervous system.[1] When the 

manifestation of anxiety significantly disrupts daily life, it is 

referred to as an anxiety disorder.[2] Some symptoms of 

anxiety are panic attacks, fear, sweating, elevated blood 

pressure, stress, tension, tremor and vomiting.[3] According to 

Freud theory, anxiety is an unpleasant state of mind. He also 

mentioned the variability between objective anxiety and 

neurotic anxiety, primarily by considering whether the origin 

of the danger stemmed from external circumstances or internal 

impulses. Objective anxiety involves an intricate internal 

response to the anticipation of injury or harm stemming from 

an external threat. Neurotic anxiety is characterized by 

feelings of apprehension and heightened physiological 

alertness.[4] If anxiety is detected early they can be treated as 

well as the symptoms can be minimized and will provide 

quality to one’s life.[5] 

Anxiety can emerge as a significant concern for 

individuals of all ages, including both adults and children. 

Studies have proved that Women tend to experience anxiety 

more frequently than men.[6] To avoid the occurrence of 

anxiety, drugs like benzodiazepines, serotonin reuptake  

Access this article online 

 

Received –  09th July 2024 

Initial Review –  28th September 2024 

Accepted – 03rd October 2024 

Quick Response Code 

inhibitors (SSRI’s) serotonin norepinephrine reuptake 

inhibitors (SNRI) are the first line drugs that are prescribed for 

treatment of anxiety. Tricyclic antidepressants, Monoamine 

oxidase inhibitor (phenelzine), antihistamines (hydroxyzine), 

anti-seizure drugs (gabapentin) are mostly prescribed as a 

second line treatment. Injecting this drug alters 

neurotransmitter levels like serotonin or GABA.[7] Prevention 

of anxiety disorder at early stage is necessarily important.[8] 

The etiology of anxiety mainly involves psychological factors 

such as childhood trauma or stressful past events; genetic 

factors also come into the considerations; physiological 

factors including alterations in serotonergic and 

catecholaminergic systems of the body play a role in causing 

of anxiety.[9] 

Due to the trends seen in those years, several authors 

called the twentieth century as "The age of anxiety". This 

description suggests that contemporary life has led to 

increased anxiety levels. In recent years, individuals appear to 

be experiencing higher levels of anxiety, with concerns about 

safety, social acceptance, and job security weighing more 

heavily on their minds than in previous times.[10] According to 

the World Health Organization (WHO), approximately 450 

million people worldwide suffer from anxiety. The current 

global prevalence rate of anxiety disorders among individuals 

________________________________________________ 

Correspondence to: Ashoka Shenoy M, Department of 

Pharmacology, Srinivas College of Pharmacy, Mangalore, 

India  

Email: shenoyscp@gmail.com 

A 

mailto:shenoyscp@gmail.com


Savvaser et al.                                                                    Anxiolytic Effects of Phyllanthus niruri Leaf Extract in Mice 

Online First                                                                                                                      Indian J Pharm Drug Studies | 2  

aged 18 years and older is 18.1%.[11] Approximately one-

eighth of the total population is affected by anxiety disorders, 

which is a heterogeneous type of condition.[12] The anxiety 

disorders are mainly classified as panic disorder, generalized 

anxiety disorder, post-traumatic stress disorder, obsessive–

compulsive disorder, social phobia, and specific phobias.[13] 

Among anxiety disorders, panic disorder (10.3%), social 

phobia (2.7%), and generalized anxiety disorder (2.2%) are 

particularly common.[14]  

The symptoms and the disorders following anxiety can be 

due to the impaired regulation of the central nervous system. 

These physical and emotional symptoms of this dysfunction 

are due to increased sympathetic stimulation at varying levels. 

Anxiety is the result of an imbalance or abnormal functioning 

of the neurotransmitters in the body. Most commonly, 

serotonergic and noradrenergic neurotransmitters play a 

prominent role in the modulatory steps involved. Disruption 

of the GABA system is also one of the actions in the 

physiology of the body due to anxiety. It is also seen that the 

corticosteroid level may increase or decrease the activity of 

various pathways in the brain.[15] 

Plants are said to be rich source of biomolecule which has 

vast therapeutic uses to cure various illnesses.[16] The synthetic 

drugs and medications contain multiple side effects; hence the 

herbal drugs that possess variety of therapeutic values are 

used in the treatment of anxiety which shows few adverse 

reactions in the body. For preparation of anxiolytic and 

antidepressant drugs from plant source requires 

multidisciplinary approach including ethanopharmacological 

surveys, careful examination of the folkloric uses of the plant, 

as well as phytochemical and pharmacological studies.[17]  

Phyllanthus niruri belongs to the family 

phyllanthaceae.[18] It is a herb that is seen growing in the 

tropical and subtropical regions of Asia, America, and China. 

Phyllanthus niruri is an annual herb that thrives in the wild, 

following the initial monsoon rains in regions such as 

Jharkhand, Bihar, Chhattisgarh, and other Indian states. There 

have been reports that this plant also grows habitually in the 

coastal regions.[19] It is an erect annual herb which grows up to 

a height of 40-70 cm with ascending herbaceous branch. The 

leaves are numerous in number; green in colour; sub sessile in 

structure; they are arranges closely.[20]  

In the Ayurvedic system of medicine in India, Chinese 

traditional medicine, and Indonesian medicine, Phyllanthus 

niruri is utilized.[21] It is rich in bioactive compounds, 

including lignans (such as phyllanthin, hypophyllanthin, and 

niranthin), flavonoids, glycosides, tannins, alkaloids, 

ellagitannins, triterpenes, phenylpropanoids, steroids, ricinolic 

acid, niruriside, and phyltetralin.Numerous reports have 

highlighted the anti-inflammatory, anti-viral, anti-cancer, and 

anxiolytic potential of various Phyllanthus species.[22] Among 

the various bioactive molecules identified from Phyllanthus 

sources, Niranthin, a lignan, has shown promise in managing 

anxiety disorders, primarily through its influence on GABA 

receptors.[23] Chlorogenic acid obtained from P. niruri, 4-

sinapoyl quinic acid not only is a powerful oxidizing agent but 

also exhibits anti-inflammatory, anticancer and anti-anxiety 

properties.[22] 

MATERIALS 

Experimental animals  

The experiment employed healthy Swiss albino mice, 

weighing between 18 to 25 grams, of either sex, which was 

obtained from the animal facility at Srinivas College of 

Pharmacy in Mangalore. These mice were kept in a controlled 

environment with a temperature of 22±2 degrees Celsius, a 

relative humidity of 60±5%, and a 12-hour light/dark cycle. 

They were accommodated in clean polypropylene cages filled 

with sterile paddy husk as bedding material, and they were 

provided with unrestricted access to a standard pellet diet and 

water. All the animals were provided with humane care in 

accordance with the guidelines established in the "Guide for 

the Care and Use of Laboratory Animals" which was 

developed by the ‘National Academy of Sciences’ and 

published by the ‘National Institute of Health’. Prior to their 

involvement in the study, the animals were allowed a 

minimum of one week for acclimatization. Furthermore, all 

procedures were conducted in compliance with the regulations 

set forth by the Institutional Animal Ethics Committee, as per 

the directives of the CPCSEA (Committee for the Purpose of 

Control and Supervision of Experiments on Animals), under 

the Ministry of Animal Welfare Division in the Government 

of India, located in New Delhi, India. 

Plant material 

The leaves of Phyllanthus niruri belonging to family 

Phyllanthaceae were collected from a local region of 

Kundapura in Udupi district on July of 2023. It was 

authenticated by Dr. Siddharaju MN, Assistant Professor and 

Research Guide, Department of Botany, University College, 

Mangalore. 

Chemicals 

Chemical such as Diazepam of pure analytical grade will be 

procured from E Merck (India) Ltd, Mumbai and all other 

chemicals of analytical grade were procured from local 

suppliers. 

METHODOLOGY 

PREPARATION OF ETHANOLIC EXTRACT OF 

Phyllanthus niruri LEAVES 

The fresh leaves of Phyllanthus niruri were collected locally 

during the season of monsoon. The collected material was 

further shade dried and powdered coarsely for the process of 

extraction. The powdered material was extracted by subjecting to 

cold maceration by using ethanol as solvent for 72 hours. In 

this procedure, 50g of the powder was soaked in 500ml of 



Savvaser et al.                                                                    Anxiolytic Effects of Phyllanthus niruri Leaf Extract in Mice 

Online First                                                                                                                      Indian J Pharm Drug Studies | 3  

ethanol, with continuous shaking using mechanical shaker. 

The resultant extract was filtered through Whatman No.1 filter 

paper and the filtrate was subjected to dryness at 50° on water 

bath.[24] 

PREPARATION OF STOCK SOLUTION OF THE 

EXTRACT FOR DOSING 

The ethanolic extract of Phyllanthus niruri was weighed and 

dissolved in required quantity of distilled water.Each time 

fresh preparation of the extract was prepared before 

administration. The extract was administered post orally at the 

constant volume of 100mg/kg and 200mg/kg for each 

animal.[25] 

EXPERIMENTAL DESIGNS 

The Swiss albino mice (22-27gms) of either sex were selected. 

The mice were divided into following groups (n=6) as 

follows:  

Group I: Vehicle control   

Group II: Standard group (Diazepam- 1mg/kg) (i.p) [26] 

Group III: Ethanolic extract of Phyllanthus niruri leaves 

(100mg/Kg) (p.o.) [25] 

Group IV: Ethanolic extract of Phyllanthus niruri leaves 

(200mg/Kg) (p.o.) [25] 

The treatment of the plant extract was given through oral 

route. All animals were pretreated for 20 days except 

diazepam treated animals. On 21st day, animals was treated, 

30 min before the evaluation. 

EXPERIMENTAL MODELS 

1. Elevated plus maze: 

Principle- This model of anxiety has been extensively used 

for evaluation of novel anxiolytic agents and to investigate 

psychological and neurochemical basis of anxiety. This test 

has been proposed for selective identification of anxiolytic 

and anxiogenic drugs. Anxiolytic compounds, by decreasing 

anxiety, increase the open arm exploration time; anxiogenic 

compounds have the opposite effect.  

Procedure- Prior to starting the experiment, the mice was 

handled daily to reduce stress. Two hours after the oral 

administration of the test drugs and 30 min after the intra-

peritoneal administration of diazepam, the animal was placed 

in the center of the maze, facing one of the open arms. 

Thereafter, the results were recorded during the next 5 min. 

An arm entry being defined when all four paws are in the arm. 

Following parameters measured: 

1. Number of open and closed arm entries. 

2. Percentage time spent in open and closed arm. 

At the end of each trial the apparatus was wiped clean in order to 

eliminate any olfactory clues, which might modify the behavior 

of next animal. The procedure was conducted preferably in a 

sound attenuated room, with observations made from an 

adjacent room via web camera attached to the computer 

system.[27] 

2. Light and dark model: 

Principle- Crawely and Goodwin (1980) Crawley (1981) 

described a simple behavior model in mice to detect 

compounds with anxiolytic effects. In a two chambered 

system, where the animals can freely move between a 

brightly-lit open field and a dark corner. Mice tends to explore 

a novel environment but toretreat from the aversive properties 

of a brightly-lit open field and a dark corner, they show more 

crossings between the two chambers and more locomotor 

activity after treatment with anxiolytic. The number of 

crossings between the light and dark sites is recorded. 

Procedure- Movements through the partition and the time 

spent in the dark and light chamber were counted. Mice were 

placed into the cage. The animals were treated 30 min before 

the experiment with test drugs or vehicle intra-peritoneally 

and then observed for 10 min, groups of 3 animals are used 

for each dose. The following behavioral were measured: 

1) The number of entries in dark and light chamber. 

2) Time spent in minutes in dark and light chambers. 

The procedure was conducted preferably in a sound attenuated 

room, with observations made from an adjacent room via web 

camera attached to the computer system.[27] 

STATISTICAL ANALYSIS 

All data were expressed as Mean ± SEM. The statistical 

significance between groups were compared using One way 

ANOVA, followed by Dunnett’s multiple comparison test. 

For all tests a "p" value of 0.05 or less was considered for 

statistical significance. 

RESULTS 

Phytochemical screening 

Phytochemical analysis is a pivotal aspect of a research, 

delves into the examination of plant-derived compounds. By 

the phytochemical screening of the Ethanolic Extract of 

Phyllanthus niruri leaves (EEPNL) it was identified that the 

extract consists of alkaloids, phenolic acids, tannins, 

flavonoids, saponins, and glycosides which helps understand 

the pharmacological activities of the plant and helps paving 

the way for the development of novel drugs, nutritional 

supplements. 

Anxiolytic effects  

In EPM (Table 1), animals treated with two doses of EEPNL 

(100mg/kg and 200 mg/kg) showed increase in the time spent 

at open arm of the elevated plus maze model which was 

significant (33.167±1.537; P<0.05 and 38.0±0.931; P<0.01) 

when compared with control (22.0±3.512). Similarly, animals 



Savvaser et al.                                                                    Anxiolytic Effects of Phyllanthus niruri Leaf Extract in Mice 

Online First                                                                                                                      Indian J Pharm Drug Studies | 4  

treated with diazepam (1mg/kg), as expected, showed a 

significantincrease in the time spent at open arm of the 

elevated plus maze model (75.0±3.651;P<0.001) 

Animals treated with two doses (100mg/kg and 200 

mg/kg) also showed decrease in the time spent at closed arm 

of the elevated plus maze model which was significant 

(109.0±16.73; P<0.01 and 76.0±11.590; P<0.001) when 

compared with control (173.33±8.33). Similarly, animals 

treated with diazepam (1 mg/kg), as expected, showed a 

significant increase in the time spent at open arm of the 

elevated plus maze model (72.00±3.425; P<0.001). 

Animals treated with two doses showed decrease in the 

number of entries in closed arm of the elevated plus maze 

model which was significant (6.833±0.60; P<0.01 and 

6.167±0.307; P<0.001) when compared with control 

(11.0±1.211). Similarly, animals treated with diazepam 

(1mg/kg), as expected, showed a significant decrease in 

number of entries at open arm of the elevated plus maze 

model (5.5±0.764; P<0.001). 

Animals also showed increase in the number of entries in 

open arm of the elevated plus maze model which was 

significant (7.83±0.477; P<0.01 and 8.83±1.249; P<0.001) 

when compared to control (4.0 ±0.516). Similarly, animals 

treated with diazepam (1mg/kg), as expected, showed a 

significant decrease in number of entries at open arm of 

elevated plus maze model(9.5±0.428; P<0.001). 

 

 

 

 

 

 

 

Fig. 1: Comparative profile of number of entries to open and 

closed arm in EPM after oral administration of 100mg/kg and 

200mg/kg of EEPNL. 

In LDT (Table 2) animals treated with two doses of 

EEPNL (100mg/kg and 200mg kg) showed reduced time 

spent in dark chamber (4.317±0.303 and 2.1±0.230;P<0.01) 

and with concomitant increase in time spent in light chamber 

(1.517±0.166; P<0.01 and 1.86±0.051; P<0.001) when 

compared with controls (4.633±0.704 and 0.650±0.141). 

Similarly, animals treated with diazepam (1mg/kg) as 

expected showed reduced the time spent in dark chamber with 

concomitant increase in time in light chamber (1.2±0.435; 

P<0.001 and 3.833±0.219; P<0.001) respectively. 

Fig. 2: Comparative profile of time spent in open and closed arm 

in EPM after oral administration of 100mg/kg and 200mg/kg of 

EEPNL. 

 

 

 

 

 

 

 

 

 

 

Fig. 3: Comparative profile of number of entries to light and 

dark chamber in LDC after oral administration of 100mg/kg and 

200mg/kg of EEPNL. 

 

Fig. 4: Comparative profile of time spent in light and dark 

chamber in LDC after oral administration of 100mg/kg and 

200mg/kg of EEPNL. 



Savvaser et al.                                                                    Anxiolytic Effects of Phyllanthus niruri Leaf Extract in Mice 

Online First                                                                                                                      Indian J Pharm Drug Studies | 5  

All animals treated two doses of EEPNL (Table 10) showed 

increased number of entries in dark chamber (8.167±.792; 

P<0.05 and 7.5±0.764; P<0.001) and with increase in number 

of entries in time in light chamber (10.0±1.15;P<0.01 and 

12.33±0.843;P<0.001) when compared with controls 

(12.0±0.577and 5.5±0.764) respectively. Similarly, animals 

treated with diazepam (1 mg/kg) as expected showed 

increased number of entries in both dark chamber and light 

chamber (3.5±0.342; P<0.001 and 15.0±1.1414; P<0.001) 

respectively. 

 

Table 1: Effect of EEPNL on Elevated Plus Maze in mice 

 

Table 2: Effect of EEPNL on light dark transition model in mice 

 

DISCUSSION 

Anxiety encompasses various dimensions, such as cognitive, 

emotional, physical, and behavioral components. 

Physiological reactions are typically normal and adaptive, but, 

they can become exaggerated to the extent that they jeopardize 

an individual's psychosocial well-being.[28] Benzodiazepines 

have been extensively used for the last 40 years to treat 

several forms of anxiety, but due to their unwanted side 

effects, alternative treatment strategies with favorable side-

effect profiles, credible benefits and moderate costs are of 

interest. Thus, there is a need to investigate alternatives.[29]In 

the present study, the anxiolytic potential of Phyllanthus 

niruri was determined using various experimental models. 

The behavioral models employed for this study were elevated 

plus maze and light and dark test. 

The EPM is considered to be an etiologically valid animal 

model of anxiety - uses natural stimuli, such as a fear of a 

new, brightly-lit open space and the fear of balancing on a 

relatively narrow raised platform, moreover it is known that 

anxiolytic agent increases the frequency of entries and time 

spent in open arm of the EPM.[29] 

Light-dark exploration test titrates a natural tendency of 

mice to explore a novel environment, against the aversive 

properties of a brightly lit compartment. Some of the 

measurable indices of anxiety are number of crossing between 

chambers, locomotor activity, number of rearing and amount 

of time spent in the dark area of the apparatus.[30] In the 

present study it is noted that administration of EEPNL 

(100mg/kg and 200mg/kg) showed dose dependent increase in 

the time spent in the open arms and the number of entries into 

open arms when compared with the control. The present study 

also showed that EEPNL (100 mg/kg and 200 mg/kg) 

increased the time spent in the light area and the no. of entries 

in to the light suggesting again that EEPNL possesses 

anxiolytic properties. 

Earlier reports on the chemical constituents of plants and 

their pharmacology suggest that plant contains alkaloids, 

flavonoids, tannins, phenolic compounds and saponins which 

Sl.no Drug 

treatment 

Dose No. of entries 

(mean±SEM) 

Time spent in seconds 

(mean±SEM) 

Open arm Closed arm Open arm Closed arm 

1 Control Vehicle 4.0 ±0.516 11.0±1.211 22.0±3.512 173.33±8.33 

2 Standard Diazepam 9.5±0.428*** 5.5±0.764*** 75.0±3.651*** 72.00±3.425*** 

3 Low dose 100mg/kg 7.83±0.477** 6.833±0.601** 33.167±1.537* 109.0±16.73** 

4 High dose 200mg/kg 8.83±1.249*** 6.167±0.307*** 38.0±0.931** 76.0±11.590*** 

Values aremean±SEM for n=6, expressed as the time (in sec) of 6 animals in each group. Data analysis was performed using 

Dunnet’s test. *P<0.05, **P<0.01, ***P<0.001 v/s control 

Sl.no Drug 

treatment 

dose No. of entries 

(mean±SEM) 

Time spent in min 

(mean±SEM) 

Light dark Light dark 

1 Control Vehicle 5.5±0.764 12.0±0.577 0.650±0.141 4.633±0.704 

2 Standard Diazepam 15.0±1.1414*** 3.5±0.342*** 3.833±0.219*** 1.2±0.435*** 

3 Low dose  100mg/kg 10.0±1.15** 8.167±.792* 1.517±0.166** 4.317±0.303 ns 

4 High dose  200mg/kg 12.33±0.843*** 7.5±0.764*** 1.86±0.051*** 2.1±0.230** 

Values aremean±SEM for n=6, expressed as the time (in min) of 6 animals in each group. Data analysis was performed using 

Dunnet’s test. *P<0.05, **P<0.01, ***P<0.001 v/s control 



Savvaser et al.                                                                    Anxiolytic Effects of Phyllanthus niruri Leaf Extract in Mice 

Online First                                                                                                                      Indian J Pharm Drug Studies | 6  

are active against anxiety and many CNS disorders in the 

body. 

Phyllanthus niruri shows anxiolytic property due to the 

presence of phenolic compounds and tannins such as 

chlorogenic acid and niranthin.[22] The phytochemical gallic 

acid shows anxiolytic activity primarily mediated by the 5-

HT1 A but not BZD receptors.[31] 

Flavonoids being structural analogues of benzodiazepines 

bind to GABAA receptor and pharmacologically act as a 

partial agonist.[29] 

The effects of EEPNL on EPM and light- dark test were 

almost equivalent to that of 1mg/kg   diazepam. In the present 

study, the anxiolytic activity of Phyllanthus niruri extract was 

observed at the dose of 200mg/kg in mice. These observations 

clearly indicate the Phyllanthus niruri exerts anxiolytic 

activity. 

It is possible that the mechanism of anxiolytic action of 

EEPNL could be mediated by synergistic action of these 

phytochemical. The results obtained in the study suggest that 

the ethanolic extract of Phyllanthus niruri possesses 

anxiolytic property. Thus, of EEPNL has potential clinic 

application in the management of anxiety disorder. Future 

investigations are warranted for elucidating the extract 

mechanism and bioactive compounds. 

CONCLUSION 

The present study can be concluded with the fact that 

ethanolic extract of Phyllanthus niruri reveals significant 

anxiolytic effect in Swiss albino mice using animal models of 

anxiety namely Elevated plus maze, light and dark chamber. 

The data obtained was satisfactory and conclusive so as to 

achieve our objectives. In the conclusion, present data indicate 

the administration of EEPNL to mice shown anxiolytic 

activity supporting the folk information regarding anxiolytic 

activity of EEPNL. 

The extract and preliminary phytochemical studies of 

EEPNL revealed the presence of chemical and phytochemical 

constituents such as (chlorogenic acid and niranthin), alkaloid, 

tannins, flavonoid, phenolic compound and saponnins. The 

exact mechanism underlying anxiolytic activity is not clear 

but it may be apparently related to active compounds present 

in the extract. Hence further studies would be necessary to 

evaluate the contribution of active chemical constituents for 

the observed anxiolytic activity. 

ACKNOWLEDGEMENTS 

The authors are grateful to management of Srinivas college of 

Pharmacy, Mangalore for providing necessary facilities to 

carry out the experiments and A.Shama Rao Foundation, 

Mangalore for providing financial assistance. 

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How to cite this article: Savvaser S, Poojary R, Habibullah 

SM, Ashoka Shenoy M. EVALUATION OF ANXIOLYTIC 

ACTIVITY OF ETHANOLIC EXTRACT OF Phyllanthus 

niruri LEAVES IN MICE. Indian J Pharm Drug Studies. 
2024; Online First. 

Funding: None;                 Conflicts of Interest: None Stated 

 

 


