INTRODUCTION:

The females comprise nearly half of the world’s population, and actively share the burdens of economy and the society. Despite this, many of them silently combat various gynaecological health issues owing to lack of awareness, and even restricted access to healthcare resources¹. While pregnancy and childbirth rightly draw focus due to their immediate risks, many non-pregnancy-related conditions—such as vulvovaginitis (VV)—are frequently overlooked. VV is commonly encountered across females of all age groups with a high prevalence^2,3. Loss of resident vaginal flora and overgrowth of transient pathogenic microbiota in the vulvovaginal region result in this condition.⁴ Although the treatment is directed to treat the causative agents (mostly, bacteria or yeast or protozoa), vaginal lubrication and intimate hygiene form the common ground^5,6.

Various herbal remedies such as tea tree oil, thyme, garlic etc. are traditionally used to treat female genitourinary tract infections⁷. In Ayurveda, the pathophysiology and management of various gynaecological disorders is elaborated under the umbrella of ‘Yonivyapada’⁸. A number of topical use formulations are described therein to be as per disease presentation and underlying causes of vulvovaginal afflictions. However, they are mostly in the form of massage oils, topical pastes, fomentations, douches etc. Although bestowed with great potency, these topical formulations have less patient compliance due to tiresome application procedures⁹.

Considering these factors, efforts were made to present the classical Ayurveda expertise in a contemporaneous manner. Vaginal gels are preferred in VV management due to their easy application and convenient packaging^10,11. Thus, an oil-based gel formulation, ‘MRUDUL’ was designed. It was developed with the herbs selected from classical Ayurveda literature indicating efficacy against VV, supported by their established pharmacological properties. Techniques of modern pharmaceutics were employed to prepare a gel dosage form. The finalized formulation was analysed based on various parameters and its standards were characterized. As bacterial and candidal VV is mostly prevalent; assessment of activity potential against these microbes was also carried out in an in-vitro study.

MATERIALS AND METHODS:

Materials:

Procurement and identification of herbal contents:

The herbal ingredients were selected on basis of their action potential against VV after a thorough literature review and discussion with experts. The list of selected ingredients is presented in (Table 1).

Table 1: Details of selected herbal ingredients

Sr. No.	Ingredient		Parts used
Sr. No.	Sanskrit Name	Latin Name	Parts used
1	Neem	Azadirachta indica A. JUSS.	Bark and Leaves
2	Lajjalu	Mimosa pudica L.	Leaves
3	Haridra	Curcuma longa L.	Rhizome
4	Yashtimadhu	Glycyrrhiza glabra L.	Root
5	Udumbara	Ficus racemosa L.	Bark
6	Triphala (Mixture of Amalaki, Bibhitaki and Haritaki)	Mixture of Emblica officinalis GAERTN., and Terminalia bellerica ROXB. and Terminalia chebula RETZ.	Fruits
7	Tila Taila	Sesamum indicum L.	Seed oil

The ingredients of formulation (except sesame oil) were procured in dry form from the authentic vendors. These were identified and authenticated at the department of Botany, SGPC’s Guru Nanak Khalsa College of Arts, Science, and Commerce, Mumbai. Organic certified black sesame oil was purchased from authentic source.

Procurement of other ingredients for gel preparation:

All the other ingredients used in gel formulation such as chemicals, excipients etc. were procured from authentic vendors in Mumbai.

Methods:

Preparation of medicated oil from the herbal ingredients:

A medicated oil (Siddha Taila) was prepared from the ingredients mentioned in Table 1, as per principles of Ayurvedic Bhaishajya Kalpana. First, a decoction (Kwatha) of certain ingredients and wet bolus (Kalka) of remaining ones were prepared. These Kwatha and Kalka were added in sesame oil and processed further (Taila Paka). The step-by-step preparation process is as follows-

Determination of quantities of raw materials for Kwatha and Kalka:

The quantities were calculated as per aforementioned general method of Taila Paka and discussion with experts. For Kwatha preparation, coarse powders were used, while Kalka was prepared with fine powders of the ingredients. The quantities are mentioned in (Table 2).

Table 2: Quantities of raw materials used in preparation of medicated oil

Ingredient	Quantity
For Kwatha
Neem	500 gm
Yashtimadhu	500 gm
Udumbara	500 gm
Triphala	500 gm
Water	16 liters
For Kalka
Lajjalu	83.3 gm
Haridra	83.3 gm

Preparation of Kwatha and Kalka:

The Kwatha was prepared with the raw materials in quantities mentioned in Table 2. As per the principles of Bhaishajya Kalpana, the process of preparation was continued till the water was reduced to ¼ of the initial quantity. Thus, 4 litres of Kwatha were obtained. The Kalka of fine powders of raw materials was prepared by adding sufficient amount of water to convert them in a wet bolus form, weighing 250gm.

Preparation of medicated oil:

According to Snehapaka procedure, 4 litres of prepared Kwatha and 250gm of Kalka were added to 1litre sesame oil. The mixture was subjected to mild heat till all the water content evaporated. The Ayurvedic confirmatory tests for medicated oils (Taila Siddhi Lakshana) were also carried out to ascertain completion of procedure. The prepared medicated oil was strained and stored in a clean container.

Phytochemical analysis and characterization of prepared medicated oil:

The prepared medicated oil was subjected to phytochemical analysis (Method- ALS/INT-MOA) and HPTLC to reveal the presence of phytoconstituents. The characterization was carried out by obtaining organoleptic (viz., colour, odour) and physico-chemical properties. The physico-chemical parameters, viz., pH (Method-IS 4309), Moisture content (Method-ISI Handbook of Food Analysis (Part XIII) – 1984), Total Ash (Method-IS 1797 – 1985), Acid insoluble ash (Method-IS 13857), Acid value (Method-ISO 660:1996), Saponification value (Method-AOAC 17th edition, 2000, Official method 920.160), Peroxide value (Method- AOAC 17th edition, 2000, Official Method 965.33), Iodine value (Method- FSSAI 02.010:2021), Ester value (Method-ISO 709:2001), Weight per ml (Method- ALS/INT-MOA), Specific gravity at 25⁰C (Method- AOAC 17th ed., 2000, Official method 920.212), Viscosity at 26.3⁰C (Method- IP 2022), and Refractive index at 20⁰C(Method- AOAC 17th ed., 2000, Official method 921.08) were assessed.

Formulation development ‘MRUDUL’ gel:

Different batches were prepared by adding different additives to the medicated oil to get a gel stable at the room temperature with no separation of oil content. The finalized formulation was subjected to further analysis.

Characterization and Evaluation of ‘MRUDUL’ gel:

The finalized gel formulation was subjected to HPTLC [CAMAG machine, Software: vision cats, Mobile phase: Toluene: Ethyl acetate: Methanol: Formic acid (5: 4: 1: 0.5)] to estimate presence of phytochemicals, in comparison with that of medicated oil. The characterization was carried out on the basis of physico-chemical parameters viz., pH, Moisture content, Total Ash, Specific gravity at 25⁰C, Viscosity at 26.3⁰C, Spreadability, Extrudability, Homogeneity, Particle size, and Consistency. Additionally, the microbial load (Method- Bacteriological Analytical Manual Chapter23: Methods for Cosmetics July 2024 Edition) with yeast and mould count was also estimated. Stability testing under accelerated conditions was carried out for a period of three months at 45⁰C temperature, and relative humidity of 75%. Organoleptic parameters, viz., appearance, aroma and form; and physico-chemical parameters viz., pH, specific gravity at 20⁰C, viscosity, spreadability, oil separation, and particle agglomeration were studied, along with total microbial count and yeast and mould count.

Assessment of antimicrobial activity using time-kill procedure:

The activity of ‘MRUDUL’ gel in reducing the microbial population of test organisms, viz., Gardnerella vaginalis (ATCC 14018) and Candida albicans (ATCC 10231), was assessed by in-vitro testing as per ASTM E2315-16 standards. In this study, the product was individually inoculated with test organisms at a concentration of approximately 10⁶ CFU/mL. Following the specified exposure period, an aliquot was withdrawn, neutralized, and subjected to the Standard Pour Plate Technique. This was done to recover the surviving microorganisms. The microbial count was determined using dilution blanks. Appropriate validation of the neutralizing agent was also performed to ensure its efficacy. The neutralizers used with their quantities were- Tween 80 (100 ml), Lecithin (30gm), Sodium Thiosulphate (5.0 gm), L histidine (1.0 gm), and Phosphate Buffer (100 ml). The contact time was 12 hours with temperature maintained at 20°C ± 1°C. The growth media used were brain heart infusion agar supplemented with 10% FBS for G. Vaginalis and Sabouraud’s Dextrose Agar for C. albicans. The assay was conducted in duplicate, and the mean colony-forming units per milliliter (CFU/mL) was calculated from the two readings.

RESULTS:

Authenticity of herbal ingredients:

The authenticity of herbal ingredients was ascertained with the help of macroscopic and microscopic identification as per API (Ayurvedic Pharmacopoeia of India) standards. They were found to be authentic.

Table 3: Physico-chemical characteristics of medicated oil

Sr. No.	Parameters	Observation	Unit
1	pH	6.5	μg/ml
2	Moisture content	0.23	%
3	Total Ash	0.2	%
4	Acid insoluble ash	0.05	%
5	Acid value	5.46	%
6	Saponification value	186	--
7	Peroxide value	2.3	mEq/Kg
8	Iodine value	103	--
9	Ester value	98.2	%
10	Weight per ml	0.95	g/ml
11	Specific gravity at 25⁰C	0.916	--
12	Viscosity at 26.3⁰C	64.7	cps
13	Refractive index at 20⁰C	1.475	--

Phytochemical analysis of medicated oil:

The phytochemical analysis of medicated oil revealed presence of Reducing sugar, Tannins, Alkaloid, Carbohydrates, Flavonoid, Glycoside, Phenol, Saponin, Steroid and Terpenoid. Starch and Proteins were absent.

Characterization of prepared medicated oil:

The results of physico- chemical characteristics are presented in (Table 3).

Development of final formulation for MRUDUL gel:

Six different batches were prepared by taking various thickening agents and excipients. The (Table 4) presents details of trial formulations and their outcomes. Of these, F6 was selected due to its favourable properties.

Table 4: Details of trial gel formulation batches

Trial Batch	Components	Quantity	Outcome
F1	Aloe vera gel	20 gm	Oil and gel separated after 24h of storage at Room temperature
F1	medicated oil	5 ml
F2	Water	100 ml	No formation of gel
	Carbopol 40	0.1 gm
	triethanolamine	0.1 ml
	medicated oil	5 ml
	emulsifier (Glyceryl stearate)	0.01 ml
F3	Water	100 ml	No formation of gel
	Carbopol 40	0.25 gm
	triethanolamine	0.25 ml
	medicated oil	5 ml
	emulsifier	0.05 ml
F4	Water	100 ml	Gel was formed; stable at room temperature; no oil separation
	Carbopol 40	0.5 gm
	triethanolamine	0.5 ml
	sodium benzoate	0.5 gm
	medicated oil	5 ml
	emulsifier (Glyceryl stearate)	0.1 ml
F5	Water	100 ml	Gel was formed; stable at room temperature; no oil separation, smoother
	Carbopol 40	0.5 gm
	triethanolamine	1.0 ml
	sodium benzoate	0.5 gm
	propylene glycol	0.1 ml
	medicated oil	5 ml
	emulsifier (Glyceryl stearate)	0.25 ml
F6	Aloe vera gel	10 gm	Gel was formed; stable at room temperature; no oil separation
	medicated oil	12.5 gm
	emulsifier (Glyceryl stearate)	0.5 ml
	sodium benzoate	0.5 gm

Characterization of gel:

Physico-chemical characterization:

The final product was a yellow color gel with characteristic odor. The results of its physicochemical properties are presented in (Table 5).

Table 5: Results for physico-chemical characterization of MRUDUL gel

Parameters	Result	Unit
pH	4.5	μg/ml
Viscosity at 26.30C	21020	Cps
Spreadability	22.8	g.cm/sec
Extrudability	Easy	--
Homogeneity	Good	--
Particle size	150	mm
Consistency	Smooth	--
Type of smear	Non- greasy	--
Phase separation	No phase separation	--
Greasiness	Non-greasy	--
Microbial load	10	cfu/ml
Yeast and mold count	--	cfu/ml

Stability Study:

The test results indicated that the gel had shelf life of 12 months, with no disturbance of physico-chemical characteristics and without microbial presence viz., total plate count and yeast and mold count. The (table 6) elaborates the stability study results.

Table 6: Results of stability test

Test parameter	3 months	6 months	9 months	12 months
Organoleptic parameters
Appearance	Yellow	Yellow	Yellow	Yellow
Aroma	Pleasant	Pleasant	Pleasant	Pleasant
Form	Smooth	Smooth	Smooth	Smooth
Physico-chemical parameters
pH	4.5	4.5	4.5	4.52
Specific gravity at 20⁰C	0.86	0.87	0.86	0.86
Viscosity (cps)	3523.1	3522	3526.2	3527.7
Spreadability (cm/s)	431.75	431.75	432.08	433.12
Oil separation	None	None	None	None
Particle agglomeration	No clumps	No clumps	No clumps	No clumps

Comparative HPTLC analysis for medicated oil and gel:

The HPTLC chromatogram were observed under visible light, and UV 366 nm before and after derivatization. The (Figure 1) represents the fingerprinting profiles., the track 1 is of medicated oil, while track 2 is for MRUDUL gel. It can be observed that the profiles are identical.

Based on their identical Rf values and peak count, both samples likely contain the same compounds. The differences in peak areas and shape might suggest quantitative differences.

Assessment of antimicrobial activity of gel:

The antimicrobial activity of MRUDUL gel was assessed against two common VV pathogens, viz., Gardnerella vaginalis and Candida albicans. The gel was absolutely effective in reducing both the microbes after 12 hours analysis. The results are presented in (Table 7).

DISCUSSION:

The discerning feature of present study was blending the practicality and usefulness of contemporary pharmaceutics with age-old wisdom backed by research evidences. VV is a commonly encountered gynaecological condition, requiring a discrete treatment due to social taboos related to women’s intimate health. Thus, a novel formulation is a convenient dosage form was developed, with affirmation of its functionality potential¹².

The selection of ingredients was based on their activity potential not just as per Ayurveda classics but contemporary researches. Neem and Haridra, the two champions from Ayurveda, are acclaimed for their antimicrobial properties^13,14. Lajjalu¹⁵ and Udumbara¹⁶ possess astringent properties and are especially elaborated in management of Yonivyapada or female reproductive system disorders. Triphala has a cleansing action and gets rid of obnoxious discharge and pruritus due to it¹⁷. In all, these main ingredients harmoniously manage the precipitating factors and presenting problems of conditions such as VV.

Rather than isolating chemical compounds from the selected herbs, we used the Ayurvedic method of medicated oil preparation. This process enables to retain both lyophilic (E.g., flavonoids, tannins) and lyophobic (E.g., terpenoids, steroids) phytochemicals. Although it could be used as it is, efforts were taken to convert it in a contemporaneous drug delivery form to increase the patient compliance and shelf life. Gels constitute a well-accepted semi-solid vaginal delivery system owing to their easy and mess-free application and great action potential^18,19.They are especially useful to achieve local pharmacological effects in conditions such as VV, where systemic medications might not prove sufficient. In the present work, a vaginal gel was developed with specially curated herbal formula, analysed to characterize its properties and evaluated for anti-microbial potential.

The formula for the ‘MRUDUL’ gel consisted of medicated oil (as active ingredient), aloe vera gel (as gelling agent/base), emulsifier (for blend the oil and aloe gel) and sodium benzoate (as preservative). Using Carbopol instead of aloe would have been a customary choice. However, aloe was preferred because it is a natural ingredient that works well as a gel base and has medicinal benefits.²⁰ The prepared formulation had an agreeable odour. Thus, to avoid chances of allergic reactions, no additional fragrances were added. The HPTLC procedure confirmed the appropriateness of procedure, as the densitogram and peak profiles were almost identical.^21,22 Acceptable physicochemical characteristics of the gel included appearance, homogeneity, and uniformity, as well as a soft, smooth texture free of excessive stickiness. During stability tests, no physical alterations or indications of phase separation were found.

LIMITATIONS OF STUDY:

The focus of present study was development, characterization and evaluation of preliminary antimicrobial activity of MRUDUL gel. The lack of in-vivo/clinical study limits understanding of other important aspects like safety, hormonal impact, vaginal flora balance, etc. To address these shortcomings, the authors intend to undertake such studies in future.

CONCLUSION:

This study demonstrated that the traditional wisdom of Ayurveda pharmacology and pharmaceutical processes can be successfully transformed into a contemporaneous dosage form. This gel formulation offers better patient compliance as it is practical, and simpler to use than an oil.In subsequent research, a clinical study would be conducted to evaluate the effectiveness of this optimised gel formulation MRUDUL to ascertain its efficacious and safe use.

CONFLICT OF INTEREST:

The authors affirm that no potential conflicts of interest exist in relation to this work.

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Received on 25.07.2025 Revised on 17.11.2025

Accepted on 27.01.2026 Published on 01.07.2026

Available online from July 04, 2026

Research J. Pharmacy and Technology. 2026;19(7):3023-3029.

DOI: 10.52711/0974-360X.2026.00430

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Test Organism	Exposure Time	Microbial Count		Antimicrobial Log Reduction	Antimicrobial Percentage Reduction*
Test Organism	Exposure Time	Count	Log	Antimicrobial Log Reduction	Antimicrobial Percentage Reduction*
Gardnerella vaginalis	Initial	1.20 x 10⁵	5.07	-	-
Gardnerella vaginalis	12 Hrs.	<10	<1	>4.07	>99.99
Candida albicans	Initial	9.60 x 10⁴	4.98	-	-
Candida albicans	12 Hrs.	<10	<1	>3.98	>99.98