Comparative Study of Anthelmintic Activity of Aqueous and Ethanolic Extract of Leaves of Centella asiatica L.

 

Jhuma Deb1*, Nilip Kanti Deb2, Doyel Mukherjee1, Soumya Saha1

1Netaji Subhas Chandra Bose Institute of Pharmacy, Tatla, Chakdaha, Nadia - 741222 West Bengal, India.

2Vidyasagara Pharmaceutical College of Education, West Bengal. India.

*Corresponding Author E-mail: jhumaniladi@gmail.com

 

ABSTRACT:

Medicinal plants have gained popularity all over the world due to their marked biological activity. Therefore, the thrive for investigating and establishing the ethnomedicinal claims is on the rise. Centella asiatica L. (Apiaceae) is one of those plants that have ethno medicinal claims against various diseases like antimicrobial, bitter stomachic, cardiotonic, anthelmintic etc. The present study is therefore aimed at validating the ethno medicinal claims. Hence in the present study comparative Anthelmintic activity of ethanolic and aqueous extract of leaves of Centella asiatica L. was carried out on Indian earthworm Pheretima posthuma against the reference drug Albendazole. The ethanolic extract caused paralysis in 5.33 min and death in 10.40 min. in 80 mg/ml concentration. The aqueous extract of Centella asiatica L. caused paralysis in 8.25 min and death in 18.31 min. in 80 mg/ml concentration.  The reference drug Albendazole at 80 mg/ml concentration has shown paralysis at 3.34 min and occurrence of death at 5.46 min. The results suggest that the active constituents responsible for the activity may be isolated and evaluated. Further studies may also be carried out using the in vivo model to establish anthelmintic activity.

 

KEYWORDS: Centella asiatica L., Pheretima posthuma, Anthelmintic Activity, Albendazole.

 

 

INTRODUCTION: 

Centella asiatica L. (Apiaceae) commonly known as Karinga in Marathi, Mandukparni in Sanskrit and Thankuni in Bengali. Itisa small, perennial herbaceous creeper which trails along the ground1. Centella asiatica is largely recognised in India as “Brain Food” since it has the power to rejuvenate the brain cells. It is described as a sacred herb in Ayurvedic system of medicine2. The growth of this herb is widespread in India, Sri Lanka, Indonesia, Malaysia, Madagascar, and other tropical and subtropical regions3-4.

 

The herb grows up to a height of 10–35cm with glabrous stem, pink striations and rooting at nodes. Leaves grows from each node of the stem which are petioled, having crenate margin, 2 - 6cm in length and 1.5 - 5cm in width. Flowering occurs from July to August4-5. The herb is very popular in Rasayana medication in Ayurvedic system of medicine and also used as medhyarasayana to cure different physiological conditions such as epilepsy, schizophrenia, and cognitive impairment, leprosy, diarrhoea, cholera, measles, jaundice, leucorrhoea and hematemesis6. Ethno medicinally this herb has various uses like antimicrobial, bitter stomachic, cardiotonic, antidote against poison, cuts, anthelmintic etc6. The inhabitants of Manipur consume the juice obtained from fresh leaves in the morning for stomach ailments. The juice of leaves mixed along with egg yolk and 1 - 2 teaspoon of honey is consumed to improve brain power1,6. People of Kanyakumari district consume whole plant to recover from fever and sun stroke. In Madhya Pradesh, leaf juice is given to children once daily for a period of one month to recover from Rickets7. It has a wide range of chemical substances which includes Betulic acid, Asiaticoside A& B, Madecassoside, Madecassic acid, Centelloside, Thankunic acid, Isothankunic acid, Isobrahmic acid, Hydrocotylin (alkaloid), Oleic acid, Linoleic acid, Linolenic acid. Centella is also rich in Vitamin C, Vitamin B1, Vitamin B2, niacin, carotene and Vitamin A8. These chemical constituents have wide therapeutic applications in areas of antimicrobial, anti-inflammatory, antispasmodic, anticancer, neuroprotective, antioxidant, and wound healing activities4.

 

MATERIALS AND METHODS:

Plant Materials:

The fresh leaves of the plant Centella asiatica L. was collected from Nadia Nursery, Chakdaha, Nadia, West Bengal during the month of March 2024 and authenticated by Dr. Suchandra Samanta Mandal, Assistant Professor, K. Bed College, Krishnanagar, West Bengal, India.

 

Chemicals and Reagents:

The chemicals, solvents and reagents used in the study were of standard analytical grade obtained from S. D Fine Chem Ltd., Mumbai and Loba Cheme, Mumbai.

 

Animals:

Adult Indian earth worm- Pheretima posthuma of about 5 - 7cm long and 0.3 - 0.4cm in width were used for this study.

 

Standard drug:

Albendazole 400mg Tablet (GlaxoSmithKline) was used as standard.

 

Preparation of extract:

The fresh leaves collected were dried thoroughly in shade condition. It was then subjected to size reduction process. 200 gm. of the fresh leaves of the plant Centella asiatica L. was defatted with petroleum ether. The marc obtained was thoroughly dried and macerated for 12 hours each with ethanol and water. The extracts obtained were dried below 40°C to obtain concentrate of the crude extract9-10.

 

Determination of anthelmintic activity:

The Anthelmintic activity was investigated on mature Indian earthworm Pheretima posthuma, which shares morphological and physiological similarities with human intestinal roundworm parasites11-13. The Anthelmintic method was carried as per the method of Pal et al. with minor modifications14-16. Ten groups each containing six earthworms of approximately equal size were released into 10 ml of desired formulation. Each group was treated with ethanol and aqueous extract along with Albendazole (Standard drug) (40, 60 and 80 mg/ml) in normal saline with 5% DMF and in vehicle alone acting as control17-19. Time for paralysis was noted when no movement could be observed with pin prick method. Time for death of individual earth worms was recorded when the worms showed no movement either by vigorous shaking or by dipping in warm water20-23.

 

RESULTS AND DISCUSSION:

Anthelmintic Activity:

The result of Anthelmintic Activity is given in (Table 1) and result plotted in (Figure 1). The result of Anthelmintic activity on earthworm Pheretima Posthuma revealed that the ethanolic and aqueous extracts at different concentration has shown paralysis effect and has caused death of earthworms. Significant Anthelmintic properties were observed in a dose dependent manner. The mean ± SEM values [statistical analysis] were calculated for the extract. The ethanolic extract of Centella asiatica L. caused paralysis in 5.33 min and death in 10.40min. in 80mg/ml concentration. The aqueous extract of Centella asiatica L. caused paralysis in 8.25min and death in 18.31min. in 80mg/ ml concentration.  The reference drug Albendazole at 80 mg/ml concentration has shown paralysis at 3.34min and occurrence of death at 5.46min.

 

Table 1: Anthelmintic activity of ethanolic and aqueous extracts of the fresh leaves of Centella asiatica L.

 

Group Treatment Dose

Reaction time in (minutes)

Time taken for paralysis (P)

Time taken for death (D)

Control

Normal Saline

-

0.00

0.00

 

Standard

 

Albendazole

40 mg/ml

7.27 ± 0.05

13.21 ± 0.22

60 mg/ml

4.16 ± 0.21

7.39 ± 0.07

80 mg/ml

3.34 ± 0.03

5.46 ± 0.11

 

 

Sample

 

Ethanol extract

40 mg/ml

10.58 ± 0.15

19.54 ± 0.23

60 mg/ml

7.38 ± 0.05

16.08 ± 0.05

80 mg/ml

5.33 ± 0.06

10.40 ± 0.03

Aqueous extract

40 mg/ml

14.32 ± 0.03

26.38 ± 0.22

60 mg/ml

10.45 ± 0.05

21.16 ± 0.17

80 mg/ml

8.25 ± 0.27

18.31 ± 0.07

Results are expressed as mean ± SEM of three observations. P – Paralysis; D – Death mg: milligram, ml: millilitre

 

Figure 1: Values of paralysis and death time of Pheretima posthuma [Indian adult earth worms] plotted against varying concentration of different extracts of fresh leaves of the plant Centella asiatica compared with standard drug (Albendazole).

 

CONCLUSION:

The medicinal plants that grow around us are immensely important since majority of them are rich in phytoconstituents which are in turn responsible for therapeutic activities. Therefore, in-depth study of medicinal plants is required. According to Ayurvedic, African and Chinese system of medicine, Centella asiatica is a very potent and ethno medicinally important plant. Literature review reveals that Centella asiatica L. has pharmacological activities like neuroprotective, antioxidant, wound healing etc.  The present study revealed prominent Anthelmintic activity in both ethanolic and aqueous extracts which is comparable to that of standard drug Albendazole. The comparative study between the ethanolic and aqueous extract revealed that the ethanolic extract has shown more promising results in comparison to the aqueous extract. The ethanolic extract has shown paralysis at 5.33min and death in 10.40min whereas the aqueous extract has shown paralysis in 8.25min and death in 18.31 min at 80mg/ml concentration which is comparable with standard drug Albendazole which has shown paralysis at 3.34min and death at 5.46min at the same concentration suggesting a need to isolate and evaluate the active constituents responsible for the exhibited biological activity.

 

CONFLICT OF INTEREST:

The authors’ Dr. Jhuma Deb, Dr. Nilip Kanti Deb, Doyel Mukherjee and Soumya Saha declare no conflict of interest, financially or otherwise.

 

ACKNOWLEDGEMENTS:

The authors are grateful to Dr. Suchandra Samanta Mandal, Assistant Professor, K. Bed College, Krishnanagar, West Bengal, India and Management of Netaji Subhas Chandra Bose Institute of Pharmacy for providing the facilities to work.

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Received on 10.07.2024      Revised on 02.11.2024

Accepted on 12.01.2025      Published on 02.08.2025

Available online from August 08, 2025

Research J. Pharmacy and Technology. 2025;18(8):3773-3776.

DOI: 10.52711/0974-360X.2025.00543

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