An In-Vitro Evaluation of Anthelmintic Activity of Malatungaw Leaves (Melasthoma malabthricum Linn.) on Roundworms (Ascaris suum)

Anthony R. Marin*, Clarence Thomas P. Melgar, John Joseph A. Deseo, Jarred F.

Gabriel,   Roqaiya G. Akbara

Department of Pharmacy, St. Dominic College of Asia Emilio Aguinaldo Hi-way, Bacoor City, Cavite

 

*Correspondence: amarin@sdca.edu.ph;

DOI: https://doi.org/10.71431/IJRPAS.2025.4805  

Article Information		Abstract
Review Article Received: 14/07/2025 Accepted: 01/08/2025 Published: 31/08/2025   Keywords Albendazole; Anthelmintic; Ascariasis; Malatungaw; Soil-transmitted helminths.		Inadequate sanitation and hygiene practices in developing countries contribute significantly to public health issues caused by soil-transmitted helminths, and parasitic worms including roundworms, whipworms, and hookworms. This research explores the potential anthelmintic activity of Melastoma malabthricum Linn., commonly known as Malatungaw, a medicinal plant used in folk medicine across tropical and subtropical regions. Despite its traditional use, scientific investigation into Malatungaw's anthelmintic properties remains limited. This study evaluates the anthelmintic activity of Malatungaw leaf extract against roundworms specifically through in-vitro assays, comparing its efficacy with the standard drug Albendazole. The leaves were collected from Romblon, Philippines, dried, macerated, and extracted using methanol. Phytochemical screening was done to verify the presence of alkaloids, flavonoids, saponins, and tannins, as means in supporting its potential as an anthelmintic agent. The extract's efficacy was tested against adult roundworms, measuring paralysis and death times. Results indicate dose-dependent anthelmintic activity, with significant differences observed between Malatungaw extract concentrations and Albendazole. The findings suggest Malatungaw leaves as a potential natural alternative for combating soil-transmitted helminth infections, specifically caused by roundworms, emphasizing the need for further research of its therapeutic potential. This research contributes to the ongoing efforts in developing sustainable solutions for parasitic infections, particularly in resource-limited settings

 

INTRODUCTION

The prevalence of inadequate sanitation and hygiene practices in developing countries has led to a significant public health issue caused by soil transmitted helminths, which are parasitic worms including roundworms, whipworms, and hookworms. These parasitic infections have detrimental effects on physical and cognitive development, leading to malnutrition and anemia, with children being particularly vulnerable. (Djuardi et al., 2021) Anthelmintic drugs are commonly used to treat soil-transmitted helminth infections, but the emergence of drug resistance and the potential adverse effects of these drugs have raised concerns. As a result, there is a growing interest in investigating natural sources as potential alternatives for anthelmintic agents. (Keiser et al., 2019) Melastoma malabthricum or colloquially known as Malatungaw or in other areas known as Malatungaw is a medicinal plant that is used by the local tribes for its various ethnomedicinal properties. It can be found in partially elevated areas in the Philippines. It can be classified as a small tree that can grow upwards to 5-10 cm that has partially red stems. CABI Compendium, (2022). Roundworms are obligate parasites that reside around the jejunum location in the small intestine, it produces a biochemical process to avoid being excreted by the body in order to thrive under the host. Contamination of this nematode can be attributed to cross-contamination from infected soil to the common orifices of the body or the consumption of infected raw meat or foods Guy, (2011). A study that was conducted by Holden-Dye & Walker, (2018) defined the term anthelmintics mainly by its usage towards the treatment and control of infection of parasites within the patient’s body and animals. The study also stated that these types of infections increase morbidity and mortality in patients and place a major economic burden on the food production industry Malatungaw leaves are known to contain various phytochemicals, including alkaloids, flavonoids, tannins, and saponins, which are potential anthelmintic agents. Despite this, the anthelmintic activity of Malatungaw leaves against soil-transmitted helminths remains understudied in existing literature

MATERIALS AND METHODS

Research Design

This exhibits the quantitative research method. It emphasizes the method used in conducting this

research, which aims to aid the researchers in evaluating the anthelmintic activity of Malatungaw leaves

extract (Melastoma malabthricum . Linn.) 

Sample Collection of Melastoma malabarthricum 

The researchers collected samples of Malatungaw leaves (Melastoma malabathricum. Linn) from the

province of Romblon. 

Extraction of Melastoma malabarthricum L. Leaves Extract 

The extraction technique used is the Decantation extraction method by using methanol as the solvent.

Briefly, samples are washed with tap water and then air dried at room temperature for 72 hours. The

procedures were done according to the methodology of Zaini, et al., (2017) with some marginal

alterations. The samples were then ground until it's in fine powder form, and each powdered sample was

then weighed approximately 30 Milligrams, 40 Milligrams, and 50 Milligrams respectively, and soaked in

95% methanol with an approximate ratio of 1: 10 (W/v) for 72 hours. Extracts were then decanted and

filtered by Whatman filter paper no.1. The filtrate was concentrated in a rotary evaporator at 40°C at 50

rpm. The extracts are dried at room temperature and kept in a freezer at 4°-8 C until further use. 

Phytochemical Screening 

Residues obtained from crude extracts are subjected to phytochemical analysis. Evaluation of plant

secondary metabolites using specified chemical tests, such as alkaloids, flavonoids, tannins, glycosides,

sugars, saponins, and steroids are examples of plant compounds.

Detection of Tannins 

For the detection of tannins, Ferric Chloride Test was performed. A 10% ferric chloride solution was used

to verify the presence of tannins within the leaf extract. The leaf extract was then diluted by a ratio of

1mL: 10mL in a beaker and was then filtered, transferred to a test tube, and followed by the addition of

the 10% ferric chloride solution, A positive indication would result in the formation of a dark blue color

within the sample Katarzyna and Godlewska, (2023). 

Other Tests for the Detection of Phytochemical Constituents 

Detection of Alkaloids 

For the detection of the presence of alkaloids, Dragendroff’s test and Mayer’s Test were performed. To

prepare the Dragendorff’s reagent, 0.07 g bismuth (III) nitrate was dissolved in a mixture of 0.80 mL of

acetic acid and 3.50 mL of water (Solution A), and 1.60 g of potassium iodide was dissolved in 4 mL of

water (Solution B). Then, equal parts of solution A and solution B were mixed to serve as the stock

solution. Afterward, 8 mL of the stock solution was mixed with 16 mL of acetic acid and 77 mL of water

to prepare Dragendorff’s reagent. One mL of Dragendorff’s reagent was added to 2 mL of the ethanolic

leaf extract along the side of the test tube. The presence of a red-orange precipitate indicates the presence

of alkaloids. Parbuntari et al., (2018) For the confirmatory test, Mayer’s test was performed. The reagent

was prepared by dissolving mercuric chloride and potassium iodide in water. Then, one mL of the

ethanolic leaf extract was mixed with a few drops of Mayer's 5 reagent. A positive indication of alkaloids

was the presence of yellow or creamy white precipitate in the solution. Kancherla et al., (2019) 

Detection of Saponins 

To detect the presence of saponins, a Froth Test was performed. The leaf extracts were diluted with a

sufficient amount of distilled water in a test tube and shaken vigorously using a vortex mixer for 15

minutes. It was then let to stand for 5 minutes to observe the formation of froth. A positive indication of

the presence of saponins was the foam persisting after 10 minutes Ugbogu, (2016).

Specimen Collection of Helminths 

The researchers collected helminths (Roundworms) from the small intestines of contaminated pigs within

the adult stage from the Imus Slaughterhouse Approximately 20 roundworms were collected in a wide

plastic container, as suggested by the professional who authenticated our specimen. Each container was

filled with pig swine blood originating from the same host where the roundworms were extracted as

means of nutritional medium in order to prolong its survivability without a host as the chosen specimens

feed not only the host tissue but also the host’s blood WHO (2023). It was then stored in a closed

environment at room temperature and was collected 8-24 hours after being extracted from contaminated

pigs by local butchers and employees of the slaughterhouse as per their SOP 6 protocol. The researchers

were not able to obtain it directly due to the agreement that was made with the operations manager of the

Imus slaughterhouse during the requesting periods.

Anthelmintic Screening 

An In-vitro anthelmintic assay was performed on the chosen land helminth samples in accordance with

the method of Ajaiyeoba et al., (2001) with minor alterations of the method. A standard drug

(Albendazole) with a concentration of 20mg/mL and different concentrations of methanolic leaf extracts

of Melasthoma malabthricum (30mg/ml, 40mg/ml and 50mg/mL) will be prepared in distilled water and

transferred into well-labelled Petri dishes. 4 adult worms of nearly identical sizes were introduced to their

respective petri-dish per concentration to be tested. Duration of paralysis and death of individual worms

was observed. Time of paralysis was determined when the specimens displayed no movement except

when vigorously shaken. The time of death was determined when the specimen displayed no movement

even after being shaken vigorously Husori et al., (2016). The three different doses along with

Albendazole were observed for the best duration of time of paralysis of the specimens, in which the time

of death or total paralysis was measured to determine the effectiveness of each dose. 3.7 

Post-Experimental Procedure (Disposal of Helminths) 

Upon completion of the experiment and collection and observation of data, the researcher transferred the

used specimens from the petri dish environment into a designated container wherein they were exposed to

albendazole (5mL) to ensure that the specimens were properly eliminated and prepared for the final

disposal. After exposure, the specimens are then sealed in their designated disposal bags and thrown

alongside other infectious waste. 

RESULT AND DISCUSSION

SOP 1: How are the leaves of Malatungaw extracted? The Leaves were extracted from their branches by

determining the quality of the leaves and segregate them to their respective piles, wherein the leaves with

optimal quality will be set aside for future use, while the rejected ones alongside the other parts of the

Malatungaw are put back into the rice sack to be disposed. After drying the leaves were then blended into

fine powder and were separated in each flask for each concentration. Once divided they were submerged

in 95% ethanol with a 1/10w/v for each concentration in the flasks and were left alone for 72 hours. After

72 hours with the solvent the macerated extracts were then filtered off and were sent to undergo rotary

evaporation. After finishing the rotary evaporation, they were then sealed off in a bioref until further use. 

SOP 2: Is there any significant difference between the 30mg/mL, 40mg/mL, and 50mg/mL of

Malatungaw ( M. malabthricum Linn.) extract in its anthelmintic activity?

 

 

 

 

Mean Score per Treatment for Time and Death and Time of Paralysis

 

SOP 3: Are there differences in the anthelmintic activity of Malatungaw leaves compared to the drug Albendazole (positive control)?

 

The table above shows the test for significant difference for time of death. This table shows comparison

of pvalue between treatments and whether the compared treatment is significantly different from each

other or not, The Table above shows the test for significant difference for time of death. For 30mg/mL

concentration of Malatungaw Leaves compared with 40mg/mL Concentration of Malatungaw leaves, the

computed p - value is 0.000 which is less than .05 alpha level. This would mean that there is a significant

difference in Terms of time of death and the null hypothesis is rejected. Hence, 40 mg/mL concentration

of Malatungaw Leaves is significantly faster in terms of killing the roundworms than 30 mg/mL

concentration of Malatungaw Leaves. For 30 mg/mL concentration of Malatungaw Leaves compared

with 50 mg/mL Concentration of Malatungaw Leaves, the computed p-value us 0.000 which is less than

.05 alpha level. This would mean that there is a significant difference in terms of Time of death and the

null hypothesis is rejected. Hence, 50 mg/mL concentration of Malatungaw Leaves is significantly faster

in terms of killing the roundworms than 30 mg/mL concentration of Malatungaw Leaves. For 30mg/mL

concentration of Malatungaw Leaves compared with Albendazole, the computed p-value is 0.365 which

is greater than .05 alpha level. This would mean that there is no significant difference in terms of time of

death and the null hypothesis is accepted. Hence, there is no significant difference in time of death

between 30 mg/mL concentration of Malatungaw Leaves and albendazole. For 40mg/mL concentration of

Malatungaw Leaves compared with 50 mg/mL concentration of Malatungaw Leaves, the computed

p-value is 0.000 which is less than.05 alpha level. This would mean that there is significant difference in

terms of time of death and the null hypothesis is rejected. Hence, 50 mg/mL concentration of Malatungaw

Leaves is significantly faster in terms of killing the roundworms than 40 mg/mL concentration of

Malatungaw Leaves. For 40mg/mL concentration of Malatungaw Leaves compared with Albendazole the

computed p-value is 0.000 which is less than .05 alpha level. This would mean that there is significant

difference in terms of time of death and the null hypothesis is rejected. Hence, 40 mg/mL concentration

of Malatungaw Leaves is significantly faster in terms of killing the roundworms than Albendazole. For 50

mg/mL concentration of Malatungaw Leaves compared with Albendazole, the computed p-value is 0.000

which is less than .05 alpha level. This would mean that there is a significant difference in terms of time

of death and the null hypothesis Is rejected. Hence,50 mg/mL concentration of Malatugnaw Leaves is

significantly faster in killing the roundworms than Albendazole.

 

Mean Score per Treatment for Time and Death and Time of Paralysis

 

The table above shows the mean score per treatment for both the time Death and the time of Paralysis in

minutes. For Time of Death, treatment with 30 mg/mL concentration of Malatungaw Leaves obtained

17.25 minutes; treatment with 40mg/mL concentration of Malatungaw Leaves obtained 5.5 Minutes;

treatment with 50 mg/mL concentration of Malatungaw Leaves obtained 3.58 minutes; and Albendazole

obtained 17.81 minutes For Time of paralysis, treatment with 30 mg/mL concentration of Malatungaw

Leaves obtained 16.43 minutes; treatment with 40mg/mL Concentration of Malatungaw Leaves obtained

4.56 minutes; treatment with 50 mg/mL concentration of Malatungaw Leaves obtained 2.78 minutes; And

albendazole obtained 16.01 minutes

 

 

 

 

 

This table shows comparison of p-value between treatments and whether the Compared treatment is

significantly different from each other or not. For 30mg/mL concentration of Malatungaw Leaves

compared with 40mg/mL concentration of Malatungaw Leaves, the computed p-value is 0.000 Which is

less than .05 alpha level. This would mean that there is significant difference in terms of time of paralysis

and the null hypothesis is rejected.Hence, 40 mg/mL concentration of Malatungaw Leaves is significantly

faster in terms of paralyzing the roundworms than 30 mg/mL concentration of Malatungaw Leaves. For

30 mg/mL concentration of Malatungaw Leaves compared with 50 mg/mL concentration of Malatungaw

Leaves, the computed p-value is 0.000 which is less than .05 alpha level. This would mean that there is

significant difference in terms of time of paralysis and the null hypothesis is rejected. Hence, 50 mg/mL

concentration of Malatungaw Leaves is significantly faster in terms of paralyzing the roundworms than

30 mg/mL concentration of Malatungaw Leaves. For 30mg/mL concentration of Malatungaw Leaves

compared with Albendazole, the computed p-value is 0.365 which is greater than .05 alpha level. This

would mean that there is no significant difference in terms of time of paralysis and the null hypothesis is

accepted. Hence, there is no significant difference in time of paralysis between 30 mg/mL concentration

of Malatungaw Leaves and Albendazole. 11 For 40mg/mL concentration of Malatungaw Leaves

compared with 50mg/mL concentration of Malatungaw Leaves, the computed p-value is 0.000 which is

less than .05 alpha level. This would mean that there is significant difference in terms of time of paralysis

and the null hypothesis is rejected.Hence, 50 mg/mL concentration of Malatungaw Leaves is significantly

faster in terms of paralyzing the roundworms than 40 mg/mL concentration of Malatungaw Leaves. For

40mg/mL concentration of Malatungaw Leaves compared with Albendazole, the computed p-value is

0.000 which is less than .05 alpha level. This would mean that there is significant difference in terms of

time of paralysis and the null hypothesis is rejected. Hence, 40 mg/mL concentration of Malatungaw

Leaves is significantly faster in terms of paralyzing the roundworms than Albendazole. For 50mg/mL

concentration of Malatungaw Leaves compared with Albendazole, the computed p-value is 0.000 which

is less than .05 alpha level. This would mean that there is significant difference in terms of time of

paralysis and the null hypothesis is rejected. Hence, 50 mg/mL concentration of Malatungaw Leaves is

significantly faster in terms of paralyzing the roundworms than Albendazole.

DISCUSSION

*For 30mg/mL concentration of Malatungaw Leaves compared with 50mg/mL concentration of

Malatungaw Leaves, the computed p-value is 0.000 which is less than .05 alpha level. This would mean

that there is significant difference in terms of time of paralysis and the null hypothesis is rejected. Hence,

* 50 mg/mL concentration of Malatungaw Leaves is significantly faster in terms of paralyzing the

roundworms than 30 mg/mL concentration of Malatungaw Leaves. For 30mg/mL concentration of

Malatungaw Leaves compared with Albendazole, the computed p-value is 0.365 which is greater than .05

alpha level. This would mean that there is no significant difference in terms of time of paralysis and the

null hypothesis is accepted. Hence, there is no significant difference in time of paralysis between 30

mg/mL concentration of Malatungaw Leaves and Albendazole. 

* For 40mg/mL concentration of Malatungaw Leaves compared with 50mg/mL concentration of

Malatungaw Leaves, the computed p-value is 0.000 which is less than .05 alpha level. This would mean

that there is significant difference in terms of time of paralysis and the null hypothesis is rejected. Hence,

50 mg/mL concentration of Malatungaw Leaves is significantly faster interms of paralyzing the

roundworms than 40 mg/mL concentration of Malatungaw Leaves. 

* For 40mg/mL concentration of Malatungaw Leaves compared with Albendazole, the computed p-value

is 0.000 which is less than .05 alpha level. This would mean that there is significant difference in terms of

time of paralysis and the null hypothesis is rejected. Hence, 40 mg/mL concentration of Malatungaw

Leaves is significantly faster in terms of paralyzing the roundworms than Albendazole. 

* For 50mg/mL concentration of Malatungaw Leaves compared with Albendazole, the computed p-value

is 0.000 which is less than .05 alpha level. This would mean that there is significant difference in terms of

time ofparalysis and the null hypothesis is rejected. Hence, 50 mg/mL concentration of Malatungaw

Leaves is significantly faster in terms of paralyzing the roundworms than Albendazole.

CONCLUSION

1. Malatungaw methanolic leaf extract exhibits anthelmintic activity at concentrations 40mg/mL

and 50mg/mL. Because of this, it could be used as a potential future reference for developing alternative

medications to anthelmintics. 

2. The Malatungaw methanolic leaf extract at a concentration of 50mg/mL has shown to be the

most effective concentration in its anthelmintic effect. 

3. The Malatungaw methanolic leaf extract at concentrations 40mg/mL and 50mg/mL is shown to

be more effective than the compared drug Albendazole based on the results.

CONFLICT OF INTEREST

Author(s) shall declare the conflict of interest before submission of manuscript for publication.

The conflict of interest as mentioned in manuscript before references section. Please note that all articles

submitted to the journal will be evaluated by turnitin for plagiarism.

ACKNOWLEDGEMENT 

The completion of this research study would not have been possible without the support and

assistance of numerous individuals and organizations whom we would like to express our gratitude to. We

would like to express our heartfelt gratitude to those who have contributed to the success of our research.

First and foremost, we extend our deepest appreciation to Prof. Anthony R. Marin, RPh., MSPharm,

Program Chair of the Department of Pharmacy, for his invaluable support and guidance. We are also

profoundly grateful to our advisor, Prof. Shalom B. Apura, for his unwavering support, insightful

feedback, and encouragement throughout this research project. To Ma'am Jen and our classmates for their

continuous support and collaboration. Your encouragement and assistance have been instrumental in our

progress. Special thanks to Mr. Romel F. Lazo, Market Supervisor IV, and Mr. Jose Mari D. Jamir, OIC of

Imus Slaughterhouse, for providing us with roundworms (scientifically known as Ascaris suum), which

were essential for our research. Lastly, we are deeply grateful to our parents for their endless support and

understanding. Your faith in us has been a constant source of motivation. Thank you all for your

contributions and support

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