Research Article

Pharmacological Evaluation of Withania coagulans Dunal

Ahmad Jamal Hasham1, Gul Jan1 and Abdullah2*

1Department of Botany, Abdul Wali Khan University Mardan, Pakistan; 2National Institute for Genomics and Advanced Biotechnology, National Agricultural Research Centre, Islamabad, Pakistan.

Received | January 27, 2026; Accepted | March 21, 2026; Published | March 28, 2026

*Correspondence | Abdullah, Department of Botany, Abdul Wali Khan University Mardan, Pakistan; Email: [email protected]

Citation | Hasham, A.J., G. Jan and Abdullah. 2026. Pharmacological evaluation of Withania coagulans dunal. Pakistan Journal of Weed Science Research, 32(1): 55-61.

DOI | https://dx.doi.org/10.17582/journal.PJWSR/2026/32.1.55.61

Keywords | Antipyretic, Analgesic, Anti-Inflammatory and Antispasmodic

Copyright: 2026 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

The word “pharmacology” comes from the Greek words “pharma” (drug) and “logy” (study). The scientific discipline of pharmacology investigates the effects of chemicals and drugs on living things (Currie, 2018). Any chemical molecule that affects a biological system, whether natural or man-made, is typically considered to be medicinal product. pharmacology has been divided into two parts pharmacodynamics, which studies how drugs affect living systems, or what the drug does to the body, and pharmacokinetics, which explains how the drug is absorbed, distributed, metabolized, and excreted, or how the body uses the drug. Both must be acknowledged in order to appropriately describe a medicine’s effects on a living thing (Rang et al., 2017). Pharmacological investigations are mostly focused on the treatment and prevention of sickness; they can evaluate the effects of chemical agents on subcellular, systemic, physiological, or behavioral processes. Medicinal plants that have been employed in traditional treatment for of thousands of years (Khan et al., 2014). A significant number of pharmaceutical chemicals come from natural materials, and many modern drugs used in medicines today have their roots in traditional medicinal plant (Patwardhan et al., 2008). The World Health Organization defines a medicinal plant as any plant that is biologically active that has therapeutic value or that is used as a raw material to make chemicals and pharmaceuticals (Karunamoorthi et al., 2013). Since the dawn of humanity, around 80% of the population has relied mostly on plants for medical care. Herbal treatments and health care products derived from widely used traditional herbs and plants are widely used, which is linked to the popularity of herbal medicine and its therapeutic purposes. Over 95% of medications in Ethiopia are made up of plants, and 70% of the people uses traditional medicine (Gebrewbet and Hndeya, 2023). It is believed that the medicinal plants are rich sources of compounds that can be used to make synthetic, non-pharmacopoeial, or pharmacopoeial drugs. In addition, these plants have played a crucial role in the global evolution of human cultures. Plants are an essential source of medication and have a big impact on world health (Sandberg and Corrigan, 2003). Herbal medications have many documented therapeutic benefits and no known harmful side effects, which has led to a greater awareness of them among the general public these days (Gupta and Keshari, 2013).

Withania coagulans Dual (Vijay et al., 2010) is an important medicinal plant that belongs to the Solanaceae family (Table 1). Withania coagulans are the species that can be found in the eastern Mediterranean region, which stretches all the way to South Asia. It can be found in much of Pakistan and India. There are other regional names for this plant, such as “Tukhmekaknaje-hindi” in Persian, “Akri” or “Puni-ke-bij” in Hindi, and “Spiubajja” in Afghan. The plant identified as W. coagulans Dunal is also called the “Indian cheese maker” or “vegetable rennet” because its fruits and leaves are utilized as a coagulant. The fruit’s pulp and husk berries, which contain withanin, are responsible for the fruit’s capacity to coagulate milk (Gupta 2012).

 

Table 1: Taxonomical classification of W. coagulans

Kingdom	Plantae
Division	Magnolipsida
Class	Magnolipsida
Order	Solanales
Family	Solanaceae
Genus	Withania
Species	W. coagulans

 

Source: Khodaei et al. 2012

 

Materials and Methods

Identification and collection of plant

For this study, aerial parts (leaves, stem, fruit and branches) of Withania coagulans was collected in October, 2023 from Palosi Payan Nowshera Khyber Pakhtunkhwa Pakistan the plant was identified taxonomically using the Flora of Pakistan.

Drying and powdering of plant

After the plant was collected, it was cleaned to get rid of any contaminants and let to air dry. Once the plant has dried, use an electric grinder to ground it into a powder.

Extraction of plant material

A conical flask containing 20g of plant powder was kept, and 200 ml of methanol was added to the powdered. To allow the flask to shake, it was wrapped with aluminum foil and put in the shaker for 72 hours. After 72 hours, the solution was filtered using Whatman filter paper in beaker. The beaker was placed in water bath at 40°C, to evaporated the methanol to get crude drug. After 7 days the methanol was evaporated and get the crude drug. The percentage yield of the extract was calculated (Ahmed et al., 2015).

Animal uses

The albino mice of weight about 28-33 g were brought from the veterinary Research Institute Peshawar. And hosted in AWKUM pharmacology laboratory in cage. Provide food and water with good environment.

Dose preparation

The methanolic extract of Withania coagulans was dissolved in distilled water with 10% DMSO to create dosing solutions for in vivo studies, using two doses of 150 mg/kg and 300 mg/kg body weight. Dose volumes were adjusted based on individual animal weights, and treatments were given orally via oral gavage.

Experiment design

Experimental animals were assigned to control and treatment groups at random, ensuring bias minimization, with each group consisting of five replicates (Table 2).

 

Table 2: Experimental design.

Group I	Control
Group II	Standard (10mg/kg)
Group III	Methanolic dose (150mg/kg)
Group IV	Methanolic dose (300mg/kg)

 

Pharmacological activity

Pharmacological activities were carried out for (antipyretic, analgesic, anti-inflammatory, and antispasmodic)

Anti-pyretic activity

Pyrexia caused by brewer’s yeast (Padhan et al., 2010) was used to assess the antipyretic efficacy. Four groups of the five albino mice each were created from the division of the mice, in order to evaluate the antipyretic effectiveness. Every mouse initial temperature was examined before yeast injection. Groups II, III, and IV mice received 20% brewer’s yeast solution. All mice’s temperatures were measured using a digital thermometer after 19 hours of yeast injection and observed high temperature. As a standard, the paracetamol solution was applied. The mice in group II were given a 10mg/kg of paracetamol solution. Methanolic extract was administered in group III and IV at doses of 150and 300 mg/kg. And check the temperature of each mouse 1 to 3 hours.

Analgesic activity

The acetic acid-induced writhing test was used to measure the analgesic activity. There were four mice groups are created. The mice in group I received an injection of one milliliter of normal saline. The mice in group’s II, III and IV ware injections of acetic acid. After injecting an acetic acid solution ten minutes later, the initial writhing was noted for ten minutes. Following the writhing, a 10mg/kg aspirin was given to the group II. Group’s III and IV mice received methanolic extract of Withania coagulant at dose 150 and 300 mg/kg, respectively. And observed the final writhing to next 10 to 20 mints (Ahmed et al., 2015).

Anti-inflammation activity

For the evolution of paw Edam use carrageenan. Mice were divided in to four group. Carrageenan ware injected to group II, III, IV mice in right paw to causes inflammation after 30 mints measured the paw volume. The group II received 10mg/kg aspirin as standard drug group III received 150mg/kg and group IV received 300mg/kg of methanolic extract (Ullah et al., 2018).

Anti-spasmodic activity

The passage of charcoal meal through the small intestine is known as the charcoal meal test. We separated the albino mice into four groups. Group I got water that was salted. The standard group II received atropine sulfate at a dose of 10 mg/kg. Methanolic extract was given to test groups III and IV at 150 mg/kg and 300 mg/kg respectively. After 30 minutes, mice in each group were given 1 milliliter of charcoal meal. The mice were killed and their intestines removed thirty minutes after they were fed the charcoal meal. The percentage of the small intestine’s total length that was covered by the charcoal meal was calculated for each group (Shamkuwar, 2021).

Statistical analysis

The experiments were performed in triplicates to avoid biological errors. The whole data parameters and their means were compared through ANOVA by following Duncan’s multiple range test (DMRT) in the SPSS software (IBM SPASS statistic 21) to determine the significant level (P < 0.05).

Results

Anti-pyretic activity

Using the brewer yeast induced pyrexia test, the anti-pyretic effect of the aerial part of Withania coagulant. The temperature of group I (control) mice were recorded (96.4±0.28) and group II (standard) was recorded in 1hour (101.21±2.38) in 2hour (99.55±2.38) and in 3hr (97.44±2.38) for group III (150mg/kg) temperature was recorded in 1hr is 101.72±1.74 in 2hr is 100.20±1.74 in 3hr is 98.71±1.74 for group IV (300mg/kg) the temperature is recoded in 1hrs is 101.55±2.38 in 2hrs (99.30±2.38) in 3hrs (97.89±2.38). The 300mg/kg show significant result than 150mg/kg (Table 3 and Figure 1).

 

Anti-inflammation activity

This activity was conduct by carrageenan induced paw edema. The volume of paw edema was measured in mm. The group I (control) mice paw volume was 10 mm and group II (standard) after carrageenan injection it showed (13± 0.67) mm and then treat with diclofenac sodium in 1hr paw volume was (12.10± 0.67) mm in 2hr is 11.5± 0.67 and in 3hr is 10.7 ± 0.67 mm. the group III (150mg/kg) after carrageenan paw volume was (12.5 ± 0.57)mm after treatment in 1hr is 12± 0.57 mm 2 and 3hr is 11.5± 0.57 mm and (11.2± 0.57) mm. the group IV show significant result as compare to other group after carrageenan the paw volume is recorded (13.5±1.10) mm then treated with 300mg/kg methanolic extract and measured the paw volume in 1hr (12.5±1.10) mm in 2hr (11.8±1.10) mm and 3hr it shows (10.9±1.10) mm (Table 4 and Figure 2).

 

Anti-spasmodic activity

In this activity, group I (control) total intestine length was measured (35.3±17.88) cm the total length of Group II (standard) was measured at (41± 22.62) cm and group III (150mg/kg ) is 40.5± 21.00 cm group IV (300mg/kg) is 39± 22.91 cm and the covered

 

Table 3: Rectal temperature of Withania coagulant

Group	DOSE	Before yeast injection (mean ± SEM)	After yeast injection (mean ± SEM)	1 hours (mean ± SEM)	2 hours (mean ± SEM)	3 hours (mean ± SEM)
GI	N/S	96.7±0.28	No yeast injection	96.4±0.28	97.1±0.28	96.8±0.28
G II	10mg/kg	96.6±0.28	103.04±2.38	101.21±2.38	99.55±2.38	97.44±2.38
G III	150mg/mg	96.6±0.27	102.69±1.74	101.72±1.74	100.20±1.74	98.71±1.74
G IV	300mg/kg	96.5±0.26	103.27±2.38	101.55±2.38	99.30±2.38	97.89±2.38

 

Table 4: Anti-inflammation of Withania coagulant

Group	Before treatment (mean ± SEM)	1hrs after (mean ± SEM)	2hrs after (mean ± SEM)	3hrs after (mean ± SEM)
control	10 ±00	10±00	10±00	10±00
standard	13 ±0.67	12.1±0.67	11.7±0.67	11.5±0.67
150mg/kg	12.5±0.57	12±0.57	11.5±0.57	11.2±0.57
300mg/kg	13.5±1.10	12.5±1.10	11.8±1.10	10.9±1.10

 

Table 5: Effect of methanolic extract against charcoal meal

Group	DOSE	Total length of intestine (mean ± SEM)	Cover length of charcoal meal (mean ± SEM)
G I	Saline water	35.3±17.88	10±17.88
G II	10mg/kg	41±22.62	9±22.62
G III	150mg/mg	40.5±21.00	10.8±21.00
G IV	300mg/kg	39±22.91	6.6±22.91

 

length of charcoal meal at group I is 10± 17.88 cm in group II is 9± 22.62 cm group III is 10.8± 21.00 and group IV measured the cover area of charcoal meals (6.6± 22.91 cm) (Table 5 and Figure 3).

 

Analgesic activity

In this activity the group I (control) showed initial (10mints) (41±4.24) writing and final writing is 35±4.24. The group II (standard) showed initial (39±12.72) writing and final writing is 21±12.72. In Group III (150mg/kg) observed initial writhing is 43± 17.67 and final is 18± 17.67 for group IV (300mg/kg) initial writhing noted (56±19.09) and final writhing is 29±19.09 for 10 mints (Table 6 and Figure 4).

 

Table 6: Effect of Withania coagulant in analgesic activity

Group	DOSE	Initial writhing (mean ± SEM)	Final writhing (mean ± SEM)
G I	Saline water	41±4.24	35±4.24
G II	10mg/kg	39±12.72	21±12.72
G III	150mg/mg	43±17.68	18±17.68
G IV	300mg/kg	56±19.09	29±19.09

 

 

Discussion

The current study investigated the medicinal properties of Withania coagulans methanolic extract, with a focus to its analgesic, antipyretic, anti-inflammatory, and anti-spasmodic effects. In this research sheds important light on the plant’s possible medicinal uses, as it has been applied for a variety of diseases in traditional healthcare (Khan et al., 2021). The methanolic extract of Withania coagulant showed a dose-dependent decrease in temperature in mice with yeast-induced pyrexia during the evaluation of its antipyretic potential. A clear dosage-dependent antipyretic effect was demonstrated by the 300 mg/kg dose, which showed a more significant drop in temperature than the 150 mg/kg dose. This is consistent with earlier research that showed how the herb is used to treat fevers in traditional medicine. Mice with paw edema caused by carrageenan were employed to test the anti-inflammatory properties. When methanolic extract was administered at doses of 150 mg/kg and 300 mg/kg, paw edema significantly decreased. The results show that the extract’s effectiveness against paw edema is dose-dependent, with the 300 mg/kg dosage having a more noticeable anti-inflammatory impact than the 150 mg/kg dosage. Withanolides which are found in the plant and have been shown to have anti-inflammatory qualities, may be the source of this anti-inflammatory effect (Gupta 2012). The charcoal meal test was used to evaluate the anti-spasmodic activity (Shamkuwar 2021). The charcoal’s intestinal transit was considerably lowered by the extract, suggesting that it may be able to relieve spasms in the abdomen and associated discomforts. The study confirmed the existence of a definite dose-dependent antispasmodic effect by finding that a 300 mg/kg dosage was more efficacious than a 150 mg/kg dosage. This study supports the traditional usage of Withania coagulant in gastrointestinal disorders. The analgesic effect was assessed using the writhing test with acetic acid (Ahmed et al., 2015). In a dose-dependent way, the extract greatly reduced the number of writhes; the 300 mg/kg dose shown the greatest efficacy. Withania coagulant bioactive components may have modulated pain pathways, which would explain its analgesic impact and support the plant’s usage in conventional pain management techniques. The research indicates that Withania coagulans exhibits significant pharmacological properties that strengthen its traditional uses (Vijay et al., 2010). The potential of methanolic extract as a natural medicinal agent is demonstrated by its antipyretic, anti-inflammatory, anti-spasmodic, and analgesic characteristics.

Conclusions and Recommendations

This research showed conclusive evidence of the pharmacological effectiveness of Withania coagulant. In experimental animals, the plant’s methanolic extract indicated excellent antipyretic, anti-inflammatory, anti-spasmodic, and analgesic properties. These results support the traditional medical application of Withania coagulant and highlight the plant potential as an effective natural treatment for a number of diseases. The study’s dose-dependent responses imply that adjusting the dosage may improve therapeutic results. It seems that bioactive substances, including withanolides, are essential for mediating these pharmacological actions. Thus, more investigation should focus on identifying these substances and clarifying their unique modes of action. The study also emphasizes the importance it is to integrate modern scientific research and conventional medical knowledge in order to find and create unique therapeutic molecules. Withania coagulant is an excellent choice for additional research and potential inclusion into modern pharmaceuticals due to its extensive history in traditional medicine and proven pharmacological effects. In conclusion, Withania coagulant has a great deal of potential as a natural medicine. Its diverse pharmacological properties both validate its conventional applications and provide opportunities for the development of new remedies for pain, fever, inflammation, and spasms. To fully appreciate this unique medicinal plant’s therapeutic potential, more study and clinical trials are needed.

Acknowledgments

The authors are grateful to the Department of Botany, Abdul Wali Khan University Mardan, Pakistan, for providing laboratory facilities and technical support to carry out this research. We also acknowledge the National Institute for Genomics and Advanced Biotechnology (NIGAB), National Agricultural Research Centre (NARC), Islamabad, Pakistan, for academic guidance and support during this study.

Novelty Statement

This study provides the first integrated evaluation of the pharmacological potential of Withania coagulans Dunal in vivo pharmacological validation. The work highlights the antipyretic, anti-inflammatory, antispasmodic, and analgesic activities of the plant extract and supports its traditional medicinal use. The integration of pharmacological assays offers new scientific evidence for the therapeutic relevance of W. coagulans.

Authors’ Contributions

Ahmad Jamal Hasham: All experiment conduct and write the original manuscript.

Gul Jan: Paper review.

Abdullah: Data analysis, interpretation of results, manuscript writing, and correspondence with the journal.

All authors have read and approved the final version of the manuscript.

Funding

This research received no external funding.

Data availability statement

The data presented in this study are available from the corresponding author upon reasonable request.

Generative AI and AI-Assisted Technology Statement

The authors declare that no generative AI and AI assisted technology was used in the creation of this manuscript.

Conflicts of interest

The authors declare no conflict of interest.

Reference

Ahmed, S., A. Naved, Khan, and S. Siddiqui. 2015. Analgesic activities of methanol extract of Terminalia chebula fruit. Pharmacol. Pharm., 6(12): p.547. https://doi.org/10.4236/pp.2015.612056

Currie, G.M. 2018. Pharmacology, part 1: introduction to pharmacology and pharmacodynamics. J. Nuclear Med. Technol., 46(2): pp.81-86. https://doi.org/10.2967/jnmt.117.199588

Gebrewbet, G.H. and A.G. Hndeya. 2023. Phytochemical screening and antibacterial activity studies on the crude leaf extract of Solanum sisymbriifolium: traditional Ethiopian medicinal plant. Advan. Gut Microb.Res., 2023: pp.1-7. https://doi.org/10.1155/2023/5525606

Gupta, P.C. 2012. Withania coagulans Dunal-an overview. Int. J. Pharmaceut. Sciences Rev. Res., 12(2): pp.68-71.

Gupta, V. and B.B. Keshari. 2013. Withania coagulans Dunal (paneer doda): A review. Int. J. Ayurved. Herbal Med., 3(5): pp.1330-1336.

Karunamoorthi, K., K. Jegajeevanram, J. Vijayalakshmi, and E. Mengistie. 2013. Traditional medicinal plants: a source of phytotherapeutic modality in resource-constrained health care settings. J. Evidence-Bas. Complemen. Alternat. Med., 18(1): 67-74. https://doi.org/10.1177/2156587212460241

Khan, H. 2014. Medicinal plants in light of history: recognized therapeutic modality. J. Evidence-Bas. Complement. Alternat. Med., 19(3): pp.216-219. https://doi.org/10.1177/2156587214533346

Khan, M.I., M. Maqsood, R.A. Saeed, A. Alam, A. Sahar, M. Kieliszek, A. Miecznikowski, H.S. Muzammil, and R.M. Aadil. 2021. Phytochemistry, food application, and therapeutic potential of the medicinal plant (Withania coagulans): A review. Molecul., 26(22): p.6881. https://doi.org/10.3390/molecules26226881

Khodaei, M., M. Jafari, and M. Noori. 2012. Remedial use of withanolides from Withania coagolans (Stocks) Dunal. Advan. Life Sci., 2(1): pp.6-19. https://doi.org/10.5923/j.als.20120201.02

Padhan, A.R., A.K. Agrahari, and A. Meher. 2010. A study on antipyretic activity of Capparis zeylanica linn. Plant methanolic extract. Int. J. Pharma. Sci. Res., 1(3): pp.169-171.

Patwardhan, B., A. Vaidya, M. Chorghade, S. Joshi. 2008. Reverse pharmacology and systems approaches for drug discovery and development. Curr. Bioact. Compd., 2008:4, 201–212. https://doi.org/10.2174/157340708786847870

Rang, H. 2017. What is Pharmacology? https://doi.org/10.1039/BK9781782621423-00001

Sandberg, F. and D. Corrigan. 2003. Natural remedies: their origins and uses. CRC Press https://doi.org/10.1201/9781420024692.

Shamkuwar, P. 2021. Evaluation of antispasmodic potential of polyherbal formulation. Asi. J. Biol. Life Sci., 9: pp.348-351 https://doi.org/10.5530/ajbls.2020.9.52.

Ullah, S., G. Jan, F. Gul, S. Khan, M. Khattak, H. Bibi, and J. Sher. 2018. Phytochemistry, anti-inflammatory and antipyretic activities of Adiantum capillus-veneris in Swiss albino mice. Int. J. Faun. Biol. Stud., 5(3): pp.19-25.

Vijay, L., K. Murti, M. Panchal, V. Gajera, and N. Joshi. 2010. Phytopharmacological properties of withania coagulans: A review. Pharmacologyonline, 3: 24–30.