Antidepressant activity of Chrysanthemum morifolium Linn in mice


Avani Shewale*, N. S. Naikwade, P. L. Ladda

Appasaheb Birnale College of Pharmacy, Sangli. Dis – Sangli, Maharashtra, India-416416.

*Corresponding Author E-mail:




Depression is the common symptom in today’s scenario. The Ethanolic extract of Chrysanthemum morifolium Linn (EECM) contains flavonoids compound which has shown therapeutic potential in neurological diseases. The study was undertaken to evaluate antidepressant activity of EECM using Despair Swim Test (DST) and Tail Suspension Test (TST). The Swiss Albino mice weighing about 20-25 gm were used. The animals were divided into 4 groups, each group comprising of 6 animals, (n=6). Group I was control received Distilled Water (10ml/kg per oral), Group II Standard Imipramine HCl (10gm/kg per oral) and Group III and IV Test group, receives EECM (250 and 500 mg/kg per oral respectively).  All drugs were administered for 10 days. The results were analysed using one way ANOVA followed by Dunnetts test, p<0.05 was considered as significant. The effect of EECM on immobility periods of mice were assessed in DST and TST. The effect of EECM was compared with that of control. The effect of 500mg/kg showed significant reduction in immobility time of mice in both DST and TST. The present study suggests that possible antidepressant activity of EECM in mice on the 500mg/kg drug administration is more than 250mg/kg.


KEYWORDS: Chrysanthemum morifolium Linn, Imipramine HCl, Despair Swim Test, Tail Suspension Test.




Depression is common, chronic, recurring disorder with some property like low cognitive and emotional reaction that imposes high expanses to patients and remedial system. Depression is common disorder with prevalence of about 15% during lifecycle and today it is considered as main reason of disability around world and is in 4th rank among 10 main reason of world load disease[1]. Depression affects not only patients but also their friends and families. Social withdrawal, lack of motivation, sexual dysfunction, sleep disorder (in 75% of patient), de­pressed mood are main symptoms of depression[2].


Depression is a significant contributor to the global burden of disease and affects people in all communities across the world. There are several types of depression. A person with major depression experiences symptoms of depression that last for more than two weeks. A person with dysthymia experiences episodes of depression that alternate with periods of feeling normal. A person with bipolar disorder, manic-depressive illness, experiences recurrent episodes of depression and extreme elation. A person with Seasonal Affective Disorder (SAD) experiences depression during the winter months, when day length is short. Although the exact cause of depression is unknown, research suggests that depression is linked to an imbalance of the neurotransmitters serotonin, norepinephrine, and dopamine in the brain[3]. Factors that may contribute to depression include heredity, stress, chronic illnesses, certain personality traits (such as low self-esteem), and hormonal changes[4].


Today depression is estimated to affect 350 million people. The World Mental Health Survey was conducted in 17 countries found that on average about 1 in 20 people reported having an episodes of depression in the previous year. The Depressive disorders often start at a young age; they reduce people’s functioning and often are recurring. For these reasons, depression is considered as the leading cause of disability worldwide in terms of total years lost due to disability. The demand for curbing depression and other mental health conditions is on the rise globally. A recent World Health Assembly called on the World Health Organization and its member states to take action in this direction[5].


Many plants have been traditional claim in the treatment of several harmful diseases still they are nothe second leading cause of premature death or disability all over the world by the year 2020. Medicinal herbs are becoming popular day by day. Both developed and developing countries have a great demand for medicinal plants due to increased identification of natural products and it is sometimes the primary source of health care available to the poor. Since old time herbal sources are useful in the treatment of many diseases[6]. As per the literature review the most medicinal plants exerted antidepressant effect through the synaptic regulation of serotonin, noradrenaline and dopamine, regulating activity of hypothalamic pituitary adrenal axis, reinforcing antioxidant defense system. Since, the medicinal plants and their active compounds can relieve depression through different pathways and hence are considered a new source to produce antidepressant.


Some of the herbs used by the folk as traditional medicine in calming mind and elevating mood are Passion flower, Lavender, Ginkgo biloba and phytoconstituents like amino acid lysine, s-adenosyl methionine, B-vitamin, choline, gamma amino butyric acid (GABA), tryptophan and other action magnesium[7].


Chrysanthemum morifolium L. is an aromatic, perennial plant producing clump of stem 25100 cm tall from rhizome[8]. Chrysanthemum indicum L. belongs to family Asteraceae[9]. The plant contains lipids, phenols, terpenoids, and flavonoids as a phytoconstituents[10].


Chrysanthemum morifolium ancient herb used in the traditional medicines showed various biological effects which are documented in various studies. They are reported to have the anti-arthritic, anti-inflammatory activity. The plant extract was also used as analgesic, antipyretic, respiratory disorder, deterioration of bone and muscle. It was also proved to be effective to inhibit the agglutination of blood platelet and promote the myocardial blood circulation and white cell phagocytosis, and therefore it was used to treat many diseases such as furuncle[11]. Furthermore, tea of Chrysanthemum morifolium Linn has been used to treat anxiety by facilitating relaxation and curing insomnia. Recently, some studies have suggested that Chrysanthemum morifolium has anti-apoptotic effects in vitro and in vivo.

Review of the plant


Plant Profile:

Chrysanthemum morifolium L:


Fig 1- Chrysanthemum morifolium L. flower


Chrysanthemum is a cosmopolitan genus, comprising about 300 species of herbs and undershrub, among which a few yield the commercial insecticide known as Pyrethrum. Several species of Chrysanthemum are ornamental and grown in gardens for their large, showy, multicolored flowers. In India, it is cultivated on a large scale only in Kashmir, though successful trials of cultivation have been reported at Kullu, Palampur, Mayurbhanj, Kumaun, Assam, Karnataka, Kerala and Kodaikanal1. Its flowers yield an important insecticide, i.e. the pyrethrins. The aerial parts of the plant used are Stem, flower and leaves specifically the tea prepared from the flowers of Chrysanthemum morifolium is been used in Korea to relieve anxiety and to enhance the mood, antioxidant and DNA damage preventive activity was found in the flower extract.


Chrysanthemum morifolium Linn is considered to be a native of China and Japan and is extensively cultivated in the Indian gardens for its ornamental multi-coloured flowers. The flower has a sharp bitter taste and is said to be stomachic and aperient. In Indo-China, the leaves are used as depur ant and prescribed in *1 migraine. A survey of the literature on Chrysanthemum species shows that these plants are the valuable sources of various polyacetylp α k -lenic compounds, sesquiterpene lactone, sesquiterpene ketone, 7 g Q monoterpenes, cam phene alcohol and flavonoid compounds.


Taxonomy of plant:

Table 1 -Taxonomy of flower Chrysanthemum morifolium L












C. morifolium

Binomial Name

Chrysanthemum morifolium L.


Chemistry            Flower:

Glycoside, chrysanthemin, on hydrolysis glucose and cyanidin, stachydrine, oil and vitamin A.


Edible Uses:

The flower heads are pickled in vinegar. Young leaves - cooked. An aromatic tea is made from the leaves, Seed.


An essential oil obtained from the plant contains chrysanthenone, this is active on the brain center affected by Parkinson's disease.



I Procurement, Authentication, Drying and Extraction of plant:


Plant (Chrysanthemum morifolium L.) was collected in month of August 2018 from Satara region (Dist - satara) Maharashtra (India). The Plant (Chrysanthemum morifolium L.) material was authenticated by Prof. M. D. Wadmare Sir Department of Botany, Smt. Kasturbai Walchand College Sangli.


II. Preparation of plant extract:

·       The powdered flower of plant (Chrysanthemum morifolium L.) was exhaustively extracted with 90 to 95% ethanol in a Soxhlet’s apparatus. 

·       The ethanolic extract was obtained, collected and concentrated at room temperature. Then the extracts were stored until use for further study.

·       Average % yield of the ethanolic extract of Chrysanthemum morifolium L. was found to be 32.1%.


III Experimental animals:

All experiments were carried out using male and female, Swiss Albino mice. Young (6-8 weeks) mice weighing around 18-24gm used in present study. The animal had free access of food and water and they were housed in natural light-dark cycle. The Animals were acclimatized for at least 5 day to the laboratory condition before behavioral experiment. Experiments were carried out between 900h and 1600h. The experimental protocol was approved by the Institutional Animal Ethics Committee (AIEC) IAEC/ABCP/02/2018-19 and care of laboratory animals was taken as per guidelines of CPCSEA, Ministry of Forest and Environment of India.  


Experimental design:

The animals were divided into 04 groups and each group contains 06 mice. Treatments were given for 10 days by oral route. Immobility period was noted on 1st and 10th day.

Groups for Forced Swim Te

Group Treatment Dose, Route of administration 1

Group I Distilled Water 10ml/kg p.o. 2

Group II Imipramine 10mg/kg p.o. 3

Group III - Ethanolic Extract of Chrysanthemum morifolium 250mg/kg p.o. 


4 Group IV - Ethanolic Extract of Chrysanthemum morifolium 500mg/kg p.o.

Evaluation of Antidepressant Activity:

Despair Swim Test:

Despair Swim Test was proposed as model to test antidepressant activity by Porsolt in 1977. Mouse was individually forced to swim in a plastic cylinder having dimensions (height: 40cm; diameter: 18cm) containing fresh water of 15cm height and maintained at 250c (± 30c). Mouse was placed in the cylinder for the first time are initially highly active, vigorously swimming in circles, trying to climb the wall or diving to the bottom. After 2-3min. activity begins to subside and to be interspersed with phases of immobility or floating of increasing length. The immobility period was recorded for 05min. After 05min. the mouse was removed and allowed to dry before being returned to their home cages.


Tail Suspension Test:

The “Tail Suspension Test” has been described by Steru et al. (1985) as a facile means of evaluating potential antidepressants. The mouse was suspended on the edge of a shelf 58cm above a table top by adhesive tape placed approximately 01cm from the tip of the tail. The duration of immobility was recorded for a period of 05 min. Mice will be considered immobile when they hang passively and completely motionless.


Estimation of Serotonin:

Preparation of Serotonin Sample:

On the last day of experiment mice were sacrificed, Whole brain was dissected out and sub cortical region (including the stratum) was separated. Weight tissue was weight and homogenized in 5ml HCL – Butanol solution for about 01min. The sample was then centrifuged for 10 min. at 2000rpm. An aliquot supernatant phase (01ml) was removed and added to centrifuge tube containing 2.5 ml heptane and 0.31ml HCL of 0.1M. After 10min. the content in centrifuge tube was centrifuged under the same condition as above in order to separate the two phases, and overlaying organic phase was discarded. The aqueous phase (0.2ml) was then taken for estimation of Serotonin.


Standard Serotonin preparation:

Serotonin standards were prepared in distilled water and HCl Butanol (1:2). Stock solution of serotonin (500µg/ml) solution was prepared by dissolving 5mg of serotonin in 10ml of Distilled water and HCl Butanol (1:2). From this stock solution 1ml standard sample was diluted to 1000ml of distilled water and HCl Butanol (0.5 µg/ml) was prepared for further process. The working standard of concentration ranging from 0.05-0.5µg/ml were prepared by transferring appropriate volume to 10 ml volumetric flask and volume was made with DW and HCl Butanol. The absorbance is read against blank at 360-470nm in Photoflurimeter.


Estimation of Serotonin:

To 0.2ml aqueous extract 0.25ml of O-Pthaldehyde reagent was added. The fluorophore was developed by heating to 100°C for 10min. After the samples reached equilibrium with the ambient temperature, readings were taken at for serotonin 360-470nm in the spectrofluorimeter.


8 Statistical Analysis and documentation of Results:

The values are expressed as Mean ± SEM for six mice in each group. The Statistical Analysis was performed using one way ANOVA followed by Dunnett’s test. (Graph pad prism version 7.04). p value < 0.05 was taken as statistically significant





Table 2: Phytochemical screening

Sr No





Test for Carbohydrates

a. Benedict’s test






Test for Alkaloids

a. Dragendroff’s test

b. Hager’s test

c. Wagner’s test













Test for Protein –

a. Millon’s test









Test for Flavonoids

a. Shinoda test

b. Sulfuric acid test

c. Lead acetate test










Test for Glycosides –

a.Killer killlani test

b. Legal’s test

c. Baljet’s test

d. Modified brontraggr’s Test












Test for Tannins –

a. Lead acetate test

b. Bromine water test

c. Pot. Dichromate test

d.FeCl3 test

e. Acetic Acid solution      














Table 3 -Effect of Ethanolic Extract of Chrysanthemum morifolium L. flower on DST

Sr. No.

Drug Treatment

Immobility time (sec.)

1st Day

10th Day


Control - Distilled water (10ml/kg, p.o.)


243.0±6.197 (2.079%)


Standard – Imipramine (10mg/kg, p.o.)

129 ±3.819****

119 ±2.028**** (8.326%)


Test I - EECM (250mg/kg, p.o.)


151 ±3.260**** (29.944%)


Test II - EECM (500mg/kg, p.o.)


130 .3±3.809**** (35.687%)

way ANOVA followed by Dunnett’s test (n = 6). ***p<0.001, ****p ˂ 0.0001 as compared to control Values were expressed in a Mean ± SEM. The results were analysed statistically by the one - group. The values in bracket indicates   that the % reduction in Immobility time.


Fig. 2- Effect of Chrysanthemum morifolium flowers extract on immobility period in Despair swim Test.


In this test animal treated with two doses of 250mg/kg and 500mg/kg showed significant **** p<0.0001 decrease in immobility time on 1st day and on the 10th day also  respectively when compared with control group on 1st day and on 10th day similarly animal treated with Imipramine (10mg/kg) showed significant decrease in immobility time.

Estimation of Serotonin in DST model

Table 4: Level of Serotonin in Brain tissue homogenate in DST model

Sr No


Concentration of serotonin in µg/gm. of wt tissue


Control - Distilled water (10ml/kg, p.o)



Standard- Imipramine (10mg/kg, P.o)

96 ±0.4216****


Test I - EECM (250mg/kg, p.o.)



Test II -EECM (500mg/kg, p.o.)



Values are expressed in a Mean ± SEM. Statistical analysis of data was carried out by one way ANOVA followed by Dunnett’s test. ****p<0.0001as compared to control group.


Fig. 3 -Level of Serotonin from brain tissue homogenate in DST model.

The neurotransmitter levels from brain tissue homogenate were also estimated. In this estimation animal treated with 2 doses of Chrysanthemum morifolium 250 mg/kg and 500 mg/kg showed significant increase in level of Serotonin ****p<0.0001 respectively when compared with control group similarly animal treated with Imipramine (10mg/kg) showed significant increase in level of Serotonin.







Evaluation of Tail Suspension Test

Table 5: Effect of Ethanolic Extract of Chrysanthemum morifolium L. flower on Tail Suspension Test:

Sr No

Drug Treatment

Immobility time (sec)

1st day

10th Day


Control - Distilled water (10ml/kg, p.o)


199.7±3.556 (18.71%)


Standard- Imipramine (10mg/kg, P.o)

130.01 ±3.525****

116.00 ±1.688**** (8.25%)


Test I – EECM (250mg/kg, p.o.)

220.01 ±2.705***

172.7±3.159**** (25.311%)


Test II - EECM (500mg/kg, p.o.)


143.3±2.140**** (35.021%)

Values were expressed in a Mean ± SEM. The results were analyzed statistically by the one - way ANOVA followed by Dunnett’s test (n = 6). ***p<0.001,****p˂ 0.0001 as compared to control group. The values in bracket indicates that the % reduction in Immobility time.


Fig. 4 - Effect of Chrysanthemum morifolium flowers extract on immobility period in Tail Suspension Test.


In this test animal treated with two doses of 250mg/kg and 500mg/kg showed significant**** p<0.0001 decrease in immobility time on 1st day and on the 10th day also respectively when compared with control group on 1st day and on 10th day similarly animal treated with Imipramine (10mg/kg) showed significant decrease in immobility time .



Estimation of Serotonin in TST model:

Table 6: Level of Serotonin in Brain tissue homogenate in TST model

Sr No

Drug Treatment

Concentration of serotonin in µg/gm of wt tissue


Control - Distilled water (10ml/kg, p.o)



Standard- Imipramine (10mg/kg, P.o)



Test I – EECM (250mg/kg, p.o.)



Test II - EECM (500mg/kg, p.o.)



Values are expressed in a Mean ± SEM. Statistical analysis of data was carried out by one way ANOVA followed by Dunnett’s test. ****p <0.0001 as compared to control group.


Figure 5: Level of Serotonin from brain tissue homogenate in TST model.

The neurotransmitter levels from brain tissue homogenate were also estimated. In this estimation animal treated with 2 doses of Chrysanthemum morifolium 250mg/kg and 500mg/kg showed significant increase in level of Serotonin ****p<0.0001 respectively when compared with control group similarly animal treated with Imipramine (10mg/kg) showed significant increase in level of Serotonin.



Depression and mental health problems in general and sensile neurological disorder in particular, are widely prevalent in modern fast paced life with the multitude of stressful life. As many as10-15% of individuals with this disorder and upto 25% of those with bipolar disorder, display suicidal behavior during the lifetime. It is currently leading one of the causes of morbidity and mortality. Despite advances in treatment and frequently used antidepressant are associated with increasing incidence of relapse adverse effect and drug interaction.


A variety of chemical and synthetic drugs are available to treat depression, but most of patients fail to tolerate adverse effects due to drugs. Moreover, only 50% of patients experience complete recovery.


Currently, studies are being increasingly conducted to detect new and economical drugs to treat depression with no adverse effect. Depression is heterogeneous mood disorder characterized with regular negative moods, feeling helplessness and is caused by decreased brain level of monoamines such as noradrenaline, dopamine, serotonin.


Since initial hypothesis of depression has been formulated long ago and proposing that symptoms of depression due to functional deficiency of cerebral monoaminergic transmitters such as Noradrenaline, serotonin and dopamine located at synapses. Imipramine hydrochloride acts by inhibiting Noradrenaline and Serotonin reuptake and has been used as a standard drug in majority studies. The beneficial effect of Imipramine hydrochloride in the forced swimming test model seems to be due to increased availability of Noradrenaline and serotonin at the post synaptic site following reuptake inhibition.


Phytochemical components especially alkaloids, saponins, flavonoids, phenols, carbohydrates have been reported to have antidepressant activity. Despair Swim Test (DST) and Tail Suspension Test (TST) are most commonly used models for the screening of the new antidepressant activity. Both the models predispose rodents to state of behavioral despair which is comparable to human depression. Both the paradigms are widely accepted behavioral model for assessing pharmacological and antidepressant activity. Immobile behavior shows lowered mood. The agents that decrease this behavior are presumed to have antidepressant effects. The same pattern of immobility was observed in present study which can be correlated with findings of Porsolt et al. in both model DST and TST.


In DST test when animal forced to swim in restricted area, initially has vigorous activity and then showed immobile posture and its movement were restricted to those movements that keep its head above water (Porsolt et al., 1977). The reduction in the immobility time was observed in the both group 250mg/kg (151 ±3.260) and 500mg/kg (130.3±3.809) but 500mg/kg group showed maximum decrease in immobility time (****p<0.0001).


The percentage decrease in immobility of Test group II 500mg/kg (35.68%) was comparable with Test group I 250mg/kg (29.66%) in DST model.


Based on hypothesis of depression monoamine such as Serotonin, Noradrenaline and Dopamine play important role in development of depression. In this investigation the level of biogenic amine are also estimated and result indicated that the level of Serotonin was increased significantly, in 500 mg/kg ethanolic flowers extract of Chrysanthemum  treated group (test group II) since  extract contain chemical constituent pyrethroids, sesquiterpenoids, flavonoids, coumarins, triterpenoids, steroids, phenolics, purines, lipids, aliphatic compounds and monoterpenoids acts as precursor of the synthesis of the biogenic amine and might be responsible for antidepressant activity. In test II (500mg/kg) group (93.50±0.4282) showed significant increase when compared to whole control group. Standard drug Imipramine (95.33±0.4216) also showed increase in level of Serotonin.


In TST, immobility reflects a state of despair which can be reduced by several agents which are therapeutically effective in human depression. This induces a state of behavioral despair in animals, which is claimed to reproduce a condition similar to human depression. 


In TST result showed that administration of Chrysanthemum morifolium produce diminution of immobility time of mice exposed to TST. In present study extracts 250mg/kg (172.7±3.159) and 500mg/kg (143.3±2.140) produce significant antidepressant effect and there efficacies were found to be comparable to standard drug Imipramine (116.00 ±1.688).


The percentage decrease in immobility was observed in TST model, the Test group II 500mg/kg showed the significant results (35.02%) as compared with Test group I 250mg/kg (25.31%).


Among treated group for Tail Suspension test model Chrysanthemum morifolium 500mg/kg group (35.02%) showed maximum decrease in immobility time (****p<0.0001) as similar to DST model (35.68%). Brains serotonin level was also increased in treated and standard group. In test group increase in serotonin level in brain as compared to control group.


The preliminary phytochemical screening indicated that the presence of Flavonoids, Alkaloids and Glycoside in flower of Chrysanthemum morifolium L, have been shown to possess anti- depressant effect.



The finding of present investigation suggests antidepressant activity of Chrysanthemum morifolium in DST and TST models of depression. Chrysanthemum morifolium L significantly reduced the immobility period in both DST and TST. Further it is found that, the group treated with 500mg/kg dose shows more significant restoration in the immobility time and increased level of brain serotonin. In the conclusion, we can say that the Ethanolic extract of flower of Chrysanthemum morifolium L are having potential to be considered as antidepressant but at the same time more extensive is required to support antidepressant action.


However, precise mechanism by which extract produced antidepressant like effect is not completely understood. Further studies would be necessary to evaluate the contribution of the chemical constituents. These chemical constituents which are active and they have observed antidepressant activity as it still remains to be determined which components are responsible for these effects.



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Received on 22.10.2019            Modified on 08.11.2019

Accepted on 16.11.2019            © A&V Publications All right reserved

Asian J. Res. Pharm. Sci. 2019; 9(4):260-266.

DOI: 10.5958/2231-5659.2019.00040.7