Formulation and Evaluation of Herbal Tablets Containing Ficus religiosa Leaf Extract for the Management of Polycystic Ovary Syndrome (PCOS)

Plant Profile

Fig No.02: Ficus Religiosa

Ficus religiosa or sacred fig is a species of fig native to the Indian Subcontinent and Indochina that belongs to Moraceae, the fig or mulberry family. It is also known as the bodhi tree, peepul tree, peepal tree, pipala tree or ashvattha tree (in India and Nepal) [6-7]. The sacred fig is considered to have a religious significance in four major religions that originated on the Indian subcontinent: Hinduism, Buddhism, Sikhism and Jainism. Hindu and Jain ascetics consider the species to be sacred and often meditate under it. Gautama Buddha is believed to have attained enlightenment under a tree of this species. The sacred fig is the state tree of the Indian states of Odisha, Bihar and Haryana [8-10].

Traditional Uses and Ethno Medicinal Significance

In traditional systems of medicine such as Ayurveda, different parts of Ficus religiosa including leaves, bark, fruits, and roots are extensively used for the treatment of various ailments. It possesses several pharmacological properties such as antidiabetic, anti-inflammatory, antioxidant, antimicrobial, and wound healing activities. Apart from its medicinal importance, Ficus religiosa holds great cultural and spiritual value [11-12]. It is revered as the Bodhi tree under which Lord Buddha attained enlightenment, making it a symbol of knowledge and spirituality [13-14]. Due to its wide range of therapeutic uses and bioactive constituents, Ficus religiosa has attracted significant attention in modern research for the development of herbal formulations and pharmaceutical applications [11].

Table No.01: Drug Profile

LITERATURE REVIEW

Murugesu et al. (2021) In 2021, Murugesu et al. reviewed phytochemistry and applications of Ficus religiosa and highlighted its suitability for modern dosage forms, including tablets. The study emphasized its antioxidant and metabolic regulatory properties, which are beneficial for formulation into standardized herbal products.

Maheshwari et al. (2023) In 2023, Maheshwari et al. conducted a systematic review focusing on phytochemistry and pharmacology of Ficus religiosa. The study highlighted the need for converting plant extracts into stable dosage forms like tablets for improved patient compliance and dose accuracy.

Rawat et al. (2024) A recent 2024 review by Rawat et al. provided updated insights into phytochemical constituents and pharmacological activities of Ficus religiosa. The authors stressed the importance of standardization and evaluation of herbal formulations, including tablets, to ensure consistent therapeutic efficacy.

Phytochemistry Letters Review (2026) A 2026 comprehensive review summarized recent advances in phytochemistry and therapeutic applications of Ficus religiosa. It emphasized that plant-based compounds can be developed into pharmaceutical formulations with proper extraction, granulation, and evaluation techniques.

General Herbal Tablet Formulation Studies (Various Authors, 2015–2022) Multiple pharmaceutical studies (2015–2022) on herbal tablet formulation techniques describe standard procedures such as wet granulation, direct compression, and evaluation parameters (hardness, friability, disintegration time). These methods are applicable to Ficus religiosa extracts for developing effective tablet formulations.

MATERIAL AND METHODOLOGY

Materials

Table No. 02: List of Chemicals used and their uses in formulation.

Equipment:  Soxhlet apparatus, Electronic balance, Heating mental, pH Meter, Hot Air Oven, Tablet Compression Machine, Hardness tester, Disintegration apparatus, Friability Tester, FT-IR

EXPERIMENTAL METHODS

Extraction and Drying of herb

Fig No. 03: Ficus Religiosa leaves powder.

Pre-formulation study of Hydro alcoholic Extract

Study of organoleptic characters

Color of extract were observed by naked eyes. Odor of extract were felled by smelling. Taste and texture were observed by touching and tasting. All tests were carried out visually.

Solubility of extract

To check out solubility Buchanania lanzen Spreng leaf extract in different solvents and find out solubility range, here 10 mg of extract was added in 10ml variety of solvents such as Methanol, Ethanol, Acetone, Chloroform and Water and solubility was observed.

Phytochemical screening of extract

A scientific process known as phytochemical screening uses analysis, inspection, extraction, and experimentation to identify different kinds of phytoconstituents that are present in different portions. It helps in the discovery of novel drugs. By using phytochemical screening, the presence of phytoconstituents such as alkaloids, flavonoids, glycosides, phenols, saponins and tannins was verified.

Table No.03: Phytochemical screening

Experimental Design:

Tablet formulations were developed using a factorial design approach. The design matrix was generated and analyzed using Design Expert DX 13.0 (Stat-Ease Inc., MN, USA).

Two formulation variables were considered as independent factors: Polyvinylpyrrolidone (X₁) and Sodium Starch Glycolate (X₂). The prepared batches were evaluated for critical quality attributes, namely hardness and disintegration time, which were selected as dependent variables.

Table No .04: Design layout of 22 factorial design.

Formulation of Tablet

Table No. 05: Composition of 22 factorial batches of Tablets.

Process of Formulation of Tablet

• Ingredients were accurately weighed and sieved through sieve no. 40 to ensure uniform particle size
• The sieved formulation ingredients were uniformly blended using a mortar and pestle
• The binder solution was prepared by dissolving Polyvinylpyrrolidon e (PVP) in a hydroalcoholic medium to obtain a 5% w/v solution, followed by gentle heating for 10–15 minutes to ensure complete dissolution.
• Granulation was carried out by gradual addition of binder solution to the powder blend with continuous mixing until a uniform damp mass was formed.
• Screening of Damp mass was performed using sieve no. 22 to obtain wet granules of uniform size.
• Drying of the wet granules was carried out in a hot air oven at 40°C for 20 minutes to obtain dry granules with suitable moisture content.
• The dried granules were subjected to sizing through sieve no. 44 to obtain granules with uniform particle size.
• Compression of the lubricated granules was carried out using a tablet compression machine to obtain tablets with an average weight of 500 mg.

Evaluation of tablets

General appearance

The general appearance and color of tablets were found by visual determination.

Weight variation test

The weight variation test was carried out to assess the uniformity of tablet weights. A total of 20 tablets were randomly selected and individually weighed using a calibrated analytical balance, and their respective weights were recorded. The average tablet weight was calculated by summing the individual weights and dividing by the total number of tablets. Each tablet weight was then compared with the calculated mean to determine the percentage deviation. The results were evaluated in accordance with pharmacopoeial specifications, which state that not more than two tablets may deviate from the average weight by more than the prescribed percentage limit, and no individual tablet should deviate by more than twice that limit.

Hardness and Thickness

The mechanical strength and dimensional uniformity of the tablets were evaluated by determining hardness and thickness. For each formulation, 20 tablets were randomly selected. Tablet hardness was measured using a Monsanto hardness tester, while thickness was determined using a Vernier caliper.

Friability Test

Friability was assessed to evaluate the resistance of tablets to mechanical stress during handling and transportation. The test was performed using a Roche friabilator. A pre-weighed sample of tablets was placed in the friabilator, which was operated at 25 rpm for 100 revolutions, allowing the tablets to fall from a height of approximately 6 inches during each rotation. After completion of the test, the tablets were dedusted and reweighed. The percentage friability was calculated, and a weight loss of less than 1% was considered acceptable for compressed tablets.

Disintegration Time

The disintegration test was conducted to determine the time required for tablets to break down into smaller particles under specified experimental conditions. The test provides an indication of the rate at which the tablet disintegrates, which is a critical parameter influencing drug release. The study was performed using a standard disintegration test apparatus, and the time required for complete disintegration of the tablets was recorded.

In-Vitro Anti-PCOS Activity:

1. 1. 1. KGN Cells were incubated at a concentration of 1 × 104 cells/mL in culture medium for 24 h at 37°C and 5% CO2.
      2. Cells were seeded at a concentration (100 μL) 104 cells/well) in 100 μL culture medium and 20, 40, 60, 80, 100 µg/mL of samples into micro plates respectively (tissue culture grade, and 96 wells).
      3. Control wells were incubated with DMSO (0.2% in PBS) and cell line. All samples were incubated in triplicate. Controls were maintained to determine the control cell survival and the percentage of live cells after culture.
      4. Cell cultures were incubated for 24 h at 37°C and 5% CO2 in CO2 incubator.
      5. After incubation, the medium was completely removed and added 20 μL of MTT reagent (5 mg/mL PBS).
      6. After addition of MTT, cells incubated for 4 h. at 37oC in CO2 incubator.
      7. Observed the wells for formazan crystal formation under microscope. The yellowish MTT was reduced to dark colored formazan by viable cells only.
      8. After removing the medium completely. Added 200 μL of DMSO (kept for 10 min) and incubate at 370C (wrapped with aluminum foil).
      9. Triplicate samples were analyzed by measuring the absorbance of each sample by a micro plate reader at a wavelength of 550 nm.

RESULT AND DISCUSSION

Pre formulation study:

Plant Authentication:

Authentication of herb was done from Shivraj College of Science, Gadhinglaj Affiliated to Shivaji University, Kolhapur by Dr. Vinayak A. Sardesai.

Organoleptic Characters:

Table No. 06: Physical properties of extract

Solubility:

Table. No. 07: Solubility of extract in different solvent

Phytochemical Screening

Fig.no.04: Phytochemical screening

Table No.08: Phytochemical screening results

Evaluation Test

Physical appearance

Fig. No.5: Physical appearance of Tablet

Table No.09: Physical Appearance of Tablet

Pre Compression evaluation of granule

Table No.10: Pre-Compression Evaluation of Granules

Evaluation of Tablet formulation

Weight variation test

Table No.11: Weight variation test of Tablet

Hardness:

Table No.12: Hardness test of Tablet

Friability:

Table No.13: Friability of Tablet

Disintegration time:

Table No.14: Disintegration Time of Tablet

In-Vitro Anti-PCOS Activity

Table No.15: Disintegration Time of Tablet

Graph No..1: MTT Assay % of Inhibition

Fig. No.06: Control Group of MTT Assay

Fig. No. 07: Extract of MTT Assay

Fig No. 08: Standard of MTT Assay