Biology, Medicine, & Natural Product Chemistry ISSN 2089-6514 (paper) 

Volume 14, Number 1, April 2025 | Pages: 239-242 | DOI: 10.14421/biomedich.2025.141.239-242 ISSN 2540-9328 (online) 
 

 

 

 

In Vivo Evaluation of Antidepressant Potential 

of Temulawak (Curcuma xanthorriza Robb.) Ethanol Extract 

in Mus musculus 

 
Raja David Virgilius1,*, Zest Lively Limbong2, Jericho L Ginting3 

1Department of Chemistry, Faculty of Mathematics and Natural Sciences, IPB University, Indonesia. 
2Department of Forestry Management, Faculty of Forestry, IPB University, Indonesia. 

3Department of Plant Protection, Faculty of Agriculture, IPB University, Indonesia. 
 

 

Corresponding author* 
stanislausraja@apps.ipb.ac.id 

 

 
Manuscript received: 18 February, 2025. Revision accepted: 20 May, 2025. Published: 23 June, 2025. 

 

 

Abstract 

 

Temulawak (Curcuma xanthorriza Roxb.) is a medicinal plant from the Zingiberceae family widely used in Indonesia for its various 

bioactives, including antiviral, antibacterial, and hepatoprotective properties. This research aims to evaluate the antidepressant potential of 

Curcuma xanthorriza ethanol extract in Mus musculus through in vivo testing. The extraction used the maceration method with 96% 

ethanol as a solvent, yielding 5.2504% extract. The primary bioactive compounds identified in the extract were terpenoids and 

curcuminoids, which are known for their neuroprotective and antidepressant effects. The antidepressant activity was assessed using the 

tail-suspension test (TST) and forced-swimming test (FST), with immobility time as the main parameter. Amitriptyline, a standard 

antidepressant, was used as a positive control, while sodium carboxymethylcellulose served as a negative control. The results showed that 

increasing doses of Curcuma xanthorriza extract significantly reduced immobility time, indicating an antidepressant-like effect. Statistical 

analysis revealed that the immobility time in mice treated with a 650 mg/Kg BW dose of Curcuma xanthorriza extract was not 

significantly different from the amitriptyline group, suggesting comparable efficacy. These findings highlight the potential of Curcuma 

xanthorriza ethanol extract as a natural antidepressant, warranting further investigation into its mechanisms and clinical applications. 

 

Keywords: Antidepressant; Curcuma xanthorriza Roxb.; Forced-swimming test Phytochemistry; Tails-suspension test. 

 

 

INTRODUCTION 
 

Temulawak (Curcuma xanthorriza Robb.) is one of the 

plants from the Zingiberaceae family. Temulawak has a 

very similar shape to turmeric. Java Island is the place 

where temulawak thrives. Indonesians tend to use 

temulawak as a food flavoring additive and herbal 

medicine. Based on research results, temulawak ethanol 

extract has many bioactivities such as antiviral, 

antibacterial, hepatoprotective, and other bioactivities 

(Syamsudin et al., 2019). The bioactivity possessed by 

temulawak ethanol extract is due to the presence of the 

chemical compounds it contains. Scientific studies 

showed that the most abundant essential phytochemicals 

obtained from the temulawak ethanol extract were 

terpenoids and curcuminoids (Zhang et al., 2014). Other 

contents possessed by temulawak ethanol extract are 

vanillin, 3-hydroxy-6-methylacetophenone, 12,13-epoxy-

xanthorrhizol, 3-hydroxy-6-methylacetophenone, and 
dehydro-6-gingeridone (Zhang et al., 2015) as well as 

several amino acids such as methionine, phenylalanine, 

tryptophan, valine, alanine, lysine, and leucine (Rahmat 

et al., 2021). 

Mental health has become a hotly debated issue, 

especially among young people. One of the mental health 

issues that can happen to anyone is depression. 

Depression is a mood disorder that causes a persistent 

feeling of sadness and loss of interest (Suma & Hasan, 

2023). Depression is one of the leading causes of 

disability, and, if left unmanaged, it can increase the risk 

of suicide. An imbalance of neurotransmitters in the 

brain such as serotonin and dopamine is the cause of 

depression. Based on data from the Global Burden of 

Disease Study, by 2020 there will be at least 322 million 

people in the world suffering from depression (Charlie et 

al., 2024). The treatment of depression cases varies 

depending on the condition of the patient ranging from 

psychological assistance to the use of antidepressant 

drugs. 

Antidepressants refer to psychotropic drugs that are 

used to treat mental illness with prominent emotional 

depression symptoms (Yunxi et al., 2023). The use of 
antidepressant drugs is closely monitored and regulated 

by the FDA. Antidepressants are the last resort for people 

with depression. Antidepressants work to relieve the 

symptoms experienced by people with depression such as 

https://doi.org/10.14421/biomedich.2025.141.239-242


 

 
 

240 Biology, Medicine, & Natural Product Chemistry 14 (1), 2025: 239-242 

 

 

anxiety, increased fatigue, impaired concentration, and 

emotional outbursts. The mechanism of action of 

antidepressant drugs varies from inhibiting monoamine 

reuptake, inhibiting monoamine breakdown, and 

increasing serotonin reuptake and release of serotonin 

and norepinephrine (Chittaranjan & Sanjay, 2010). 
 
 

MATERIAL AND METHODS 
 

Tools and Materials 

The tools used are laboratory glassware, stative, syringe, 

analytical balance, pestle and mortar, yarn, feeding tube, 

beaker glass, spatula, and rotary evaporator.  

The ingredients used in this research were Curcuma 

xanthorriza Roxb. rhizomes, ethanol 96%, amitriptyline, 

water, and sodium carboxymethyl cellulose. 

 

Procedures 

Preparation and acclimatization of the test animals 

Animal acclimatization was carried out for 7 days by 

giving the mice standard feed and drinking water ad 

libitum. Feed is given as much as 4 grams/head/day. The 

mice used for treatment were 48 mice (n=8 head/group). 

Mice were weighed and the drum was separated in each 

treatment. All research procedures were approved by the 

Ethics Committee of the Faculty of Mathematics and 

Natural Sciences, IPB University. 
 

Preparation of Curcuma xanthorriza ethanol extract 

The temulawak rhizome was cleaned, peeled, and sliced 

before being dried in an ovent at 45⁰C for 6-8 hours. 

After drying, the sample was ground into powder and the 

simplisia was filtered using an 80-mesh sieve to obtain 

softer temulawak powder, which was extracted using the 

maceration method. For the extraction process, 100 

grams of temulawak powder was extracted using ethanol 

96% for 24 hours. The macerate was filtered using 

Whatman filter paper. Subsequently, the extracted 

solution was separated from the solvent using a rotary 

evaporator 45⁰C and 80 cmHg pressure to obtain a 

concentrated extract. The extract yield was calculated 

and stored in a vial bottle at -20⁰C to 4⁰C for measuring 

antidepressant bioactivity.  
 

Antidepressant activity test 

Antidepressant activity testing with the tail suspension 

test (TST) and forced swimming test (FST) method was 

performed based on the procedure of Castagne et al., 

2010. Mice that have been acclimated for 14 days were 

randomly divided into 4 groups, as follows the following: 

 
Table 1. Test animal grouping data. 
 

Group name Treatment 

Normal Untreated mice 

Negative control Sodium carboxymethylcellulose 0,5% 

Positive control Amitriptyline 

Group I Dose 325 mg/kg BW 

Group II Dose 650 mg/kg BW 

Mice were placed in the experimental room at least 

60 minutes before starting the experiment. The decoction 

and positive control were administered once a day. 

Antidepressant activity testing was performed 1 hour 

after oral administration of the treatment (Castagne et al., 

2010). Immobility time was observed, namely the 

duration of time not moving (body, legs, and hands in a 

state of rest) for ≥ 1 second per mouse for 6 minutes 

(Ueno et al., 2022) 

 

Data analysis 

The immobility time data were used to calculate the 

percent of antidepressant activity of the ethanol extract of 

Curcuma xanthorriza against the positive control 

amitriptyline. statistical analysis using SPSS 29.0 for 

Windows. Values expressed as mean ± SD were then 

compared with a one-way ANOVA test followed by an 

LSD post hoc test to find out whether the differences 

between treatment groups if the data are significantly 

different (H0 is rejected if the probability <0,05). 

 

 

RESULTS AND DISCUSSION 
 

Curcuma xanthorriza Roxb extract 

Curcuma xanthorriza Roxb. extraction was done by the 

maceration method by soaking Curcuma xanthorriza 

Roxb. powder in 96% ethanol solvent for 24 hours. The 

ethanol solvent was chosen because it is a polar solvent 

and can dissolve polar compounds such as terpenoids and 

curcuminoids (Aisyah & Erminawati, 2022). Terpenoids 

and curcuminoids have been shown to have 

antidepressant bioactivity. The extraction process 

produced a paste of Curcuma xanthorriza extract. A total 

of 5.2504 grams of Curcuma xanthorriza extract paste 

was produced from 100 grams of Curcuma xanthorriza 

powder. The yield of Curcuma xanthorriza maceration 

extraction was 5.2504% as shown in Table 1. The yield 

of a extraction process is influenced by several factor 

such as temperature, solute and solvent ratio, target 

compounds to be extracted, and solvent purity (Abin et 

al., 2024). Terpenoid and curcuminoid isolates have been 

shown to have antidepressant bioactivity. The yield value 

of 5.2504% obtained in this research is not much 

different from the yield value of 5.42% obtained in the 

research of Lidvina et al., 2017. 

 
Table 2. Calculation of yield from Curcuma xanthorriza extracts. 

 

Weight of C. xanthorriza 

powder 

Weight of 

extract 
%Yield 

100 grams 5.2504 grams 5,2504% 

 

Antidepressant activity of Curcuma xanthorriza 

ethanol extract 

Bioactivity testing was carried out by open tails-

suspension test (TST) and forced-swimming test (FST) 

methods on each group of mice with the measurement 



 

 
 

 Virgilius et al. – In Vivo Evaluation of Antidepressant Potential … 241 
 

 

variable being immobility time. The antidepressant 

bioactivity of Curcuma xanthorriza ethanol extract can 

be described by the immobility time. The increase in 

immobility time is directly proportional to the 

antidepressant bioactivity. Stress in mice causes 

immobility in the mice’s bodies, so a high immobility 

time indicates a high stress level. Amitriptyline was used 

as the positive control in this research. Amitriptyline was 

chosen as the positive control because amitriptyline has 

been commonly consumed as an antidepressant drug 

commonly used in the community. The mechanism of 

action of the drug amitriptyline is by inhibiting the 

reuptake of serotonin and norepinephrine at the pre-

synapse cell membrane resulting in an increase in the 

concentration of serotonin and or norepinephrine in the 

central nervous system (Kim, 2017). 

Based on Table 3, it can be seen through tails-

suspension test (TST) and forced-swiming test (FST) 

untreated mice have the highest immobility time. Mice 

given amitriptyline have the lowest immobility time 

because the stress generated can be reduced by the 

presence of amitriptyline as an antidepressant. Sodium 

carboxymethylcellulose was used as a negative control  

in this research. Sodium carboxymethylcellulose is 

commonly used as a suspending agent in drug 

preparations (Henny et al., 2019). Based on Table 3, it 

can be seen that increasing the dose of Curcuma 

xanthorriza decreases the immobility time so that it can 

be seen that there is linearity between increasing the dose 

of Curcuma xanthorriza extract and decreasing the 

immobility time. Based on the results of the least 

significant difference (LSD) test, it can be seen that there 

is no significant difference between amitriptyline and a 

dose of 650 mg/Kg BW of Curcuma xanthorriza extract.  

 

 

 

Table 3. Antidepressant bioactivity testing result. 

 

Group Body weight (g) TST Immobility Times (s) FST Immobility Times (s) 

Untreated mice 2.87 ± 0.35 155.43 ± 12.42 173.48 ± 12.42 

Na-carboxy methyl cellulose 3.19 ± 0.58 135.00 ± 16.90 145.71 ± 11.58 

Amitriptyline 2.31 ± 0.59 79.75 ± 21.02 75.57 ± 16.76 

325 mg/Kg BW dose of C. xanthorriza 2.10 ± 0.29 102.75 ± 13.47 103.86 ± 14.16 

650 mg/Kg BW dose of C.xanthorriza 2.04 ± 0.53 80.01 ± 17.47 77.81 ± 11.98 

 
 

 

CONCLUSIONS 
 

Ethanol can be used to extract Curcuma xanthorriza 

using the maceration method. Curcuminoids and 

terpenoids can be extracted using ethanol. The yield of 

Curcuma xanthorriza ethanol extract was 5.2504%. 

Antidepressant bioactivity testing can be done by tails-

suspension test and also forced-swim test. The increase 

in Curcuma xanthorriza ethanol extract levels will be 

inversely proportional to the immobility time of the mice. 

Mice injected with amitriptyline had an immobility time 

in the tails-suspension test of 80.01±17.47s and in the 

forced-swimming test of 75.57±16.76s. The 650 mg/Kg 

dose has immobility time test result that are not 

significantly different from amitriptyline, namely in the 

tails-suspension test of 80.01±17.47 and 77.81±11.98 s in 

the forced-swimming test. 

 

 

Acknowledgements: The author acknowledges the 

Department of Chemistry, Faculty of Mathematics and 

Natural Science, IPB University for providing material 

support for the research.  

 

Authors’ Contributions: Raja David Virgilius conceived 

the research design, conducted the antidepressant 

bioactivity testing, analyzed the data, and wrote the 

manuscript. Zest Lively Limbong dan Jericho L Ginting 

planted, cared for, and harvested Curcuma xanthorriza 

Robb. and conducted Curcuma xanthorriza Robb. 

extraction. All author read and approved the final version 

of the manuscript. 

 

Competing Interests: The authors declare that there are 

no competing interests. 

 

Funding: The author declares that this research is funded 

by the Ministry of education, culture, research, and 

higher education (Kemendikbud-ristekdikti RI) through 

the Program Kreativitas Mahasiswa (PKM) 2023 funding 

program.  

 

 

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