INTRODUCTION:

Ficus racemosa L. is a tropical fig species widely distributed across South and Southeast Asia, including Indonesia, where it occurs naturally but remains underutilized, primarily cultivated as an ornamental or bonsai plant. Phytochemical investigations have confirmed that Ficus racemosa L fruit contains diverse classes of secondary metabolites such as alkaloids, phenols, flavonoids, tannins, and terpenoids, which contribute to a broad spectrum of pharmacological properties, particularly antioxidant activity^1,2.

Flavonoids, one of the major subclasses of plant phenolics, are well recognized for their potent antioxidant potential. By donating electrons or hydrogen atoms, they effectively neutralize reactive oxygen species (ROS) and protect cellular components from oxidative damage³. Their antioxidant activity is commonly evaluated using in vitro radical scavenging assays such as DPPH, ABTS, and FRAP, with the DPPH assay being widely applied owing to its simplicity, reproducibility, and sensitivity^4,5. Given the increasing demand for safe and sustainable antioxidants, flavonoid-rich extracts are of considerable interest for pharmaceutical and cosmetic applications.

Extraction efficiency plays a critical role in maximizing flavonoid recovery from plant matrices. Conventional techniques such as maceration and soxhlet extraction are widely applied because of their simplicity and low cost; however, they often require prolonged extraction times, consume large volumes of organic solvents, and may yield incomplete recovery of target compounds^6,7. By contrast, Microwave-Assisted Extraction (MAE) has emerged as an advanced, non-conventional technique that employs dielectric heating to enhance solvent penetration and mass transfer, resulting in higher efficiency, shorter extraction times, reduced solvent consumption, and improved selectivity for thermolabile phenolic compounds^8,9. Optimization of operational parameters, however, remains essential to prevent degradation of sensitive metabolites¹⁰.

Beyond their therapeutic significance, flavonoid-rich extracts are increasingly explored as natural ingredients in cosmetic formulations, particularly sunscreens. Sunscreens are designed to prevent ultraviolet (UV)-induced skin damage, including erythema, photoaging, and carcinogenesis¹¹. The Sun Protection Factor (SPF) is an internationally accepted parameter that quantifies the photoprotective efficacy of formulations. Recent studies have demonstrated that phenolic-rich plant extracts, such as those derived from Camellia sinensis (green tea), Vitis vinifera (grape seed), and Curcuma longa (turmeric), not only provide strong antioxidant activity but also enhance SPF values when incorporated into topical formulations¹². Such comparisons emphasize the importance of evaluating less-studied botanicals like Ficus racemosa L within the broader context of natural antioxidants with photoprotective potential.

Previous investigations of Ficus racemosa L have mainly centered on crude extracts obtained via conventional techniques. Methanolic extracts of leaves and fruits exhibited antioxidant and antibacterial activities¹³, ethyl acetate root extracts demonstrated high phenolic content and significant DPPH scavenging activity¹⁴, and butanolic fruit fractions showed notable antioxidant and hypoglycemic effects¹⁵. While these findings underscore the pharmacological value of the plant, no systematic studies have applied MAE to Ficus racemosa L fruit in combination with solvent fractionation, quantitative flavonoid profiling, and evaluation of its potential as a sunscreen ingredient.

The novelty of the present study lies in its integrated and application-oriented approach. An optimized MAE protocol was employed to enhance flavonoid recovery from Ficus racemosa L fruit, followed by liquid–liquid fractionation to enrich bioactive compounds. The fractions were comprehensively analyzed for total flavonoid content and antioxidant capacity using the DPPH assay, then incorporated into cream formulations for sunscreen applications. To establish translational value, physicochemical characteristics, irritation safety, and in vitro SPF assessments were systematically performed. By bridging phytochemical research with formulation science and situating racemosa L within the broader landscape of natural antioxidants, this study highlights its potential as a novel and eco-friendly source for the development of antioxidant-enriched sunscreen products.

MATERIAL AND METHODS:

Plant Material:

This study's sample is Ficus racemosa L. fruit, which was collected using purposive sampling without comparing it to the same plant from other regions. The samples were collected from the Pante Bahagia Village in Aceh (GPS coordinates: Latitude: 4.943054, Longitude: 97.173640).

Chemical and Reagents:

The chemical substances used in the study included distilled water, quercetin, sulfuric acid, hydrochloric acid from Mallinckrodt, USA; ferric chloride, ethyl acetate, chloroform, ethanol 70%, magnesium powder, amyl alcohol, Mayer’s reagent, Bouchardat reagent, Dragendorff’s reagent from Mitra Kimia, Indonesia; n-hexane, AlCl₃ (aluminum chloride) 2%, 120 mM potassium acetate, FeCl₃ (ferric chloride), from Merck, Germany; DPPH (2,2-diphenyl-1picrylhydrazyl) from TCI, Japan; and Vitamin C.

Methods:

Extraction and Fractionation:

The extraction was performed using Microwave-Assisted Extraction (MAE) with ethanol as the solvent. The process was carried out at a microwave power of 400 W for 10 minutes, conditions optimized to improve extraction efficiency while maintaining the stability of thermolabile compounds. The extract was then concentrated and subjected to liquid–liquid extraction. Further separation was performed using thin-layer chromatography (TLC) with GF254 silica gel plates as the stationary phase and chloroform:methanol:ethyl acetate:water (80:12:6:2) as the mobile phase. The ethanol extract and its fractions (n-hexane, ethyl acetate, and water) from Ficus racemosa L. fruit were subsequently tested for antioxidant activity using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay with UV-Vis spectrophotometry^16-19.

Antioxidant Testing with DPPH Solution:

Antioxidant testing was carried out in several steps: preparation of DPPH stock solution I (1000 ppm), stock solution II (100 ppm), and a working solution (40 ppm). A stock solution of Ficus racemosa L. fruit extract (1000 ppm) was prepared, followed by serial dilutions to obtain concentrations of 20, 40, 60, 80, and 100 ppm. Quercetin was used as a standard at concentrations of 1, 2, 3, 4, and 5 ppm. The maximum wavelength was determined, absorbance of the DPPH solution was measured, and the IC50 value was calculated to evaluate antioxidant activity²⁰.

Cream Formulation Procedure:

The cream was formulated into four variants: one blank (cream base) and three containing Ficus racemosa L. fruit ethanol extract at concentrations of 5%, 7.5%, and 10%. The mortar and pestle were first soaked in hot water, and all ingredients were weighed accurately. Ingredients were divided into two groups: the oil phase (stearic acid and cetyl alcohol), which was melted in a water bath at 70°C (Mass I), and the water phase (nipagin, nipasol, tea, and propylene glycol), which was dissolved in hot water (Mass II). After drying, the oil phase was combined with the water phase and homogenized thoroughly using mechanical mixing (replacing the less precise description of grinding with a mortar and pestle) until a uniform cream mass was formed²¹.

Evaluation of Cream Formulation:

The physical quality of the cream was evaluated using several parameters: organoleptic properties, homogeneity, pH, cream type, spreadability, adhesion, irritation testing on volunteers, viscosity, and stability. A cycling stability test was also conducted by subjecting the cream to repeated temperature changes (cold and warm cycles) to simulate real-world storage and usage conditions. This test provides insight into the long-term stability of the formulation under environmental stress²².

Determination of SPF Value:

Each cream formulation and a commercial sunscreen cream (positive control) was weighed (0.1 g), dissolved in 25 mL of 70% ethanol, and mixed until homogeneous, resulting in a solution concentration of 4000 ppm. The spectrophotometer was set to measure absorbance between 290–320 nm at 5 nm intervals, with 96% ethanol as the blank. Absorbance values were recorded, and the SPF values were calculated using Mansyur’s equation²³.

Statistical Analysis:

All experimental data were obtained in triplicate and expressed as mean ± standard deviation (SD). Statistical analysis was carried out using one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test to evaluate significant differences between formulations (F0–F3) in terms of antioxidant activity (IC₅₀ values), Sun Protection Factor (SPF), and physicochemical characteristics (pH, viscosity, spreadability, and adhesion). A p-value less than 0.05 (p < 0.05) was considered statistically significant. Statistical analyses were performed using SPSS version 26.0 (IBM Corp., Armonk, NY, USA).

RESULT AND DISCUSSION:

Yield Results of Ethanol Extract of Ficus racemosa L.:

The results of Ficus racemosa L. simplicia powder were extracted using a Microwave Assisted Extraction (MAE) device, filtered, and then evaporated using a rotary evaporator at a temperature of 50°C and steamed over a water bath until it produced a thick extract to evaporate the solvent so that you get a thick extract of Ficus racemosa L.

Simplicia powder Ficus racemosa L. was taken as 500 g of Ficus racemosa L. simplicia with a drying loss of 10% simplicia, 75.5 grams of Ficus racemosa L. ethanol extract with a Ficus racemosa L.extract yield of 15.1%

Determination of Total Flavonoid Content of Ficus racemosa L.:

Determination of the Maximum Wavelength of Quercetin:

The maximum wavelength of quercetin was determined at 438 nm by analyzing its solution within the 400–800 nm range. Quantitative analysis of Ficus racemosa L. ethanol extract was conducted using UV-Vis spectrophotometry, leveraging the strong absorption properties of flavonoids in the UV-visible spectrum²⁴.

A standard quercetin solution was prepared at concentrations of 4, 6, 8, 10, and 12 ppm, with corresponding absorbance values of 0.273, 0.406, 0.550, 0.688, and 0.801, respectively. The regression equation obtained was y = 0.0669x + 0.0084, with a correlation coefficient (r = 0.999), indicating a strong linear relationship between concentration and absorbance. For total flavonoid quantification, the ethanol extract was mixed with 10% AlCl₃ and 1M sodium acetate, leading to a wavelength shift in the visible spectrum and a yellowish color formation. The sample was incubated for 30 minutes to optimize the reaction and enhance color intensity²⁵. The mean absorbance of the ethanol extract measured 0.375 nm, within the Lambert-Beer law range (0.2–0.8 absorbance). The total flavonoid content was 2.1939 mg QE/g extract, where QE denotes Quercetin Equivalent, corresponding to 0.2193%, significantly lower than the 14.92% reported from maceration extraction²⁶.

Fractionation Results of Ethanol Extract of Ficus racemosa L.:

The fractionation of the ethanol extract of Ficus racemosa L. resulted in different yields depending on the solvent used. The n-hexane fraction yielded 2.98 grams, the ethyl acetate fraction produced 3.94 grams, and the water fraction obtained was 18.75 grams. The water fraction had the highest yield compared to the other fractions, which can be attributed to the predominance of polar compounds in Ficus racemosa L. The variation in yield among the different solvents is due to their differing abilities to extract specific compounds from the ethanol extract. These results highlight the influence of solvent polarity on the fractionation process, with water being the most effective in extracting the majority of compounds present in Ficus racemosa L²⁷.

Thin Layer Chromatography:

The separation of the water fraction by TLC yields an Rf value of 0.22 for the initial spot. The second stain possesses an Rf value of 0.44. The third stain possesses an Rf value of 0.73. The ethyl acetate fraction yields an initial stain Rf value of 0.15. The second stain possesses an Rf value of 0.20. The third stain yielded an Rf value of 0.51. The fourth stain exhibited an Rf value of 0.73, while the quercetin comparison displayed an Rf value of 0.71. The four discernible stains, with the fourth stain exhibiting an Rf value of 0.73, are presumed to include flavonoids, as the quercetin comparison yields an Rf value of 0.71. Simultaneously, the n-hexane fraction exhibited no discoloration. This is believed to be due to the presence of several polar and semi-polar bioactive chemicals in the Ficus racemosa L. samples, in contrast to nonpolar bioactive compounds²⁸.

The Rf value and stain color are presented in the table. Upon irradiation with a UV light at a wavelength of 254 nm and subsequent application of FeCl3 reagent, the coloration of the spots intensifies due to the formation of a complex between flavonoid compounds and Fe3+ ions in FeCl3, resulting in a darker (black) hue. The initial stain yields a dark hue (black). The second stain yields a dark hue. The final stain yields a dark hue. The fourth stain is of a dark hue²⁹.

Antioxidant Activity Analysis of Ficus racemosa L. Using DPPH Method:

The antioxidant activity of the water, ethyl acetate, and n-hexane fractions of Ficus racemosa L. was evaluated using the DPPH (1,1-diphenyl-2-picrylhydrazyl) method via UV-Vis spectrophotometry and compared with quercetin as a reference standard. Prior to testing, the maximum absorption wavelength of the DPPH solution was determined ^30,31.

The measurement results showed that a 40 ppm DPPH solution in 70% ethanol exhibited peak absorption at 518 nm, which falls within the visible light spectrum. This wavelength corresponds to the highest absorbance, ensuring high sensitivity and linearity in measurements, where small concentration changes significantly affect absorbance values.

Antioxidant test data indicated that as concentration increased, absorbance decreased, aligning with the Lambert-Beer law. This confirms the reliability of the DPPH method in assessing the antioxidant potential of Ficus racemosa L. fractions³².

Table 2: Percent value of DPPH immersion in the n-hexane fraction of Ficus racemosa L.

Test solution concentration (ppm)	Measurement			% inhibition			Average % inhibition
Test solution concentration (ppm)	1	2	3	1	2	3	Average % inhibition
Blank	0.918	0.918	0.918	0	0	0	0
100	0.874	0.860	0.859	4.793	6.318	6.427	5.846
200	0.828	0.828	0.827	9.803	9.803	9.912	9.839
300	0.760	0.760	0.759	17.211	17.211	17.320	17.247
400	0.694	0.693	0.693	24.400	24.509	24.509	24.472
500	0.658	0.658	0.654	28.322	28.322	28.758	28.467

Table 3: Mark Percent DPPH Soaking in Ethyl Acetate Fraction of Ficus racemosa L.

Test Solution Concentration (ppm)	Measurement			% soaking			Average % immersion
Test Solution Concentration (ppm)	1	2	3	1	2	3	Average % immersion
Blank	0.921	0.921	0.921	0	0	0	0
4	0,774	0.775	0.771	15.860	15.852	16.032	15.924
8	0.646	0.641	0.634	29.858	30.401	31.161	30.473
12	0.535	0.533	0.532	41.910	42.128	42.236	42.091
16	0.454	0.453	0.450	50.705	50.814	51.140	50.886
20	0.343	0,344	0.342	62.757	62.649	62.866	62.757

Table 4: Mark Percent DPPH Soaking in Ficus racemosa L.Water Fraction

Test Solution Concentration (ppm)	Measurement			% inhibition			Average % inhibition
Test Solution Concentration (ppm)	1	2	3	1	2	3	Average % inhibition
Blank	0.906	0.906	0.906	0	0	0	0
20	0.745	0.744	0.745	17.770	17.880	17.770	17.806
40	0.674	0.674	0.682	25.607	25.607	24.724	25.312
60	0.577	0.576	0.575	36.313	36.423	36.534	36.423
80	0.486	0.486	0.487	46.357	46.357	46.247	46.320
100	0.327	0,322	0.323	63.907	64.459	64.348	64.238

Results of IC50 Value Analysis of N-Hexane, Ethyl Acetate and Water Fractions of Ficus racemosa L. and Quercetin Comparator Using the DPPH Method:

The absorbance findings are analyzed to determine the Inhibition Concentration of 50% (IC50), which signifies the substrate concentration required to diminish 50% of DPPH free radical activity. The IC50 values were obtained via a linear regression equation created by graphing the concentration of the test solution versus the percentage reduction of DPPH, with concentration (ppm) on the x-axis and percentage reduction on the y-axis. Table 1 displays the IC50 values obtained from the quercetin assay and the comparator solutions.

Table 6: Results of the Regression Equation and IC50 Values from the DPPH Method Quercetin Test and Comparative Samples

Test Solution	Regression Equations	IC50 value	Activity category
N-Hexane Fraction	Y = -0.39725 + 0.058833X	856.61 ± 0.90 ppm	Inactive
Ethyl Acetate Fraction	Y = 3.0735 + 2.9535X	15.88 ± 0.65 ppm	Very Strong
Water Fraction	Y = 0.596921 + 1.83712X	26.89 ± 0.90 ppm	Very Strong
Quercetin Comparator	Y = 12.805742 + 1.399478X	26.57 ± 0.50 ppm	Very Strong

Legend:

IC₅₀ represents the concentration required to inhibit 50% of DPPH radicals; lower IC₅₀ values indicate stronger antioxidant activity. Quercetin was used as a positive control for comparison.

Molyneux (2004) stated that antioxidant activity is inversely related to IC₅₀ values, such that a lower IC₅₀ corresponds to higher radical scavenging activity. Table 3 presents the IC₅₀ values obtained in this study. Among the tested samples, the ethyl acetate fraction exhibited the strongest radical scavenging activity with an IC₅₀ of 15.88 ± 0.65 ppm, followed by the crude extract and the aqueous fraction IC₅₀ = 26.89 ± 0.90 ppm. Quercetin, used as the positive control, showed comparable activity with an IC₅₀ of 26.57 ± 0.50 ppm. In contrast, the n-hexane fraction displayed negligible activity with an IC₅₀ of 856.61 ± 0.90 ppm, thus classified as inactive. This weak performance is likely attributable to the non-polar nature of the fraction, which limits the extraction of phenolic and flavonoid compounds that are primarily responsible for antioxidant activity. These findings indicate that the ethyl acetate fraction provides superior radical scavenging potential compared to the n-hexane fraction, crude Ficus racemosa L. extract, and even quercetin, underscoring its role as the most potent antioxidant fraction.

Statistical analysis demonstrated that the IC₅₀ values differed significantly among fractions (p < 0.05), with the ethyl acetate fraction showing the strongest radical scavenging activity compared to the aqueous and n-hexane fractions³⁹.

Evaluation of Sunscreen Cream Preparations:

Organoleptic Test of Sunscreen Cream:

An organoleptic test was conducted to evaluate the sensory characteristics of different sunscreen cream formulations, including color, odor, and texture. The results indicated that the base formula (F0) exhibited a white color, a characteristic odor, and a semi-solid consistency. In contrast, formulations F1, F2, and F3 displayed a light brown hue while maintaining the same characteristic odor and semi-solid texture as F0. These findings suggest that the addition of specific ingredients in formulations F1, F2, and F3 influenced the color of the product, whereas the odor and texture remained consistent across all tested formulations.

Homogeneity Test Results:

A homogeneity test was performed on the four sunscreen cream formulations using a glass slide to observe their uniformity. The test was conducted in three repetitions, and the results confirmed that all formulations were homogeneously blended³⁹. Significant differences in viscosity were observed across formulations (p < 0.05), confirming the effect of extract concentration on rheological behavior.

pH Test Results of Cream Preparations:

The pH test of the sunscreen cream containing Ficus racemosa L. ethanol extract was conducted using a calibrated pH meter. Each formulation underwent three repeated measurements to ensure accuracy. The results showed variation in pH values across different formulations. The base formula (F0) exhibited an average pH of 6.4, while Formula F1 had a slightly lower pH of 6.0. Formula F2 recorded an average pH of 5.3, and Formula F3 had the lowest pH at 4.8. Overall, the pH values of all formulations fell within the acceptable range of 4.5 to 8, indicating their suitability for topical application³⁸. ANOVA results confirmed significant differences in pH across the formulations (p < 0.05), indicating that extract concentration influenced the acidity of the cream while remaining within the acceptable topical range.

Spreadability Test Results:

The spreadability test results for the cream formulations, conducted before the cycling test, demonstrated that all formulations met the required spreadability range of 5–7 cm. Each formulation displayed consistent and adequate spreadability, confirming their ease of application. After undergoing the cycling test, the spreadability remained within the acceptable range, indicating that the formulations maintained their consistency and stability despite temperature variations. The slight changes observed before and after the cycling test suggest that the formulations retained their original characteristics without significant alterations. Overall, these findings confirm that the cream formulations exhibit good spreadability, ensuring their effectiveness and ease of use. The spreadability values of the creams were significantly different (p < 0.05), yet all remained within the acceptable range (5–7 cm).

Adhesion Test Results:

The adhesion test was conducted to determine the duration for which the cream adhered to the skin, a crucial factor for effective absorption of active ingredients. The results indicated that the cream had an average adhesion time of 30.55 seconds with a standard deviation of 2.09 seconds. A longer adhesion time is preferable, as it enhances the absorption of active substances. Given that the minimum standard for cream adhesion is 4 seconds, the tested formulations significantly exceeded this threshold, demonstrating excellent adhesive properties that ensure prolonged contact with the skin for optimal absorption and efficacy.

Viscosity Test Results:

Viscosity testing assessed the consistency and stability of Ficus racemosa L. sunscreen formulations before and after the cycling test. Initially, F3 (10%) showed the lowest viscosity (9,025 cS), while F1 (5%), F2 (7.5%), and F0 recorded 26,414 cS, 11,867 cS, and 21,253 cS, respectively. After cycling, all formulations exhibited reduced viscosities (F3: 6,122 cS; F2: 9,126 cS; F1: 15,171 cS; F0: 11,169 cS) due to temperature fluctuations but remained within acceptable ranges, confirming stability and integrity. Adhesion times also differed significantly (p < 0.05), with all formulations exceeding the minimum 4-second requirement, supporting their suitability for topical application.

Cream Type Test Results:

The cream type was evaluated using the methylene blue dye test, where dye dispersion indicates an oil-in-water (O/W) emulsion, while lack of dispersion denotes a water-in-oil (W/O) type. All formulations (F0–F3) showed positive dye dispersion, confirming an O/W emulsion. This emulsion type is desirable for topical use due to better spreadability, absorption, and non-greasy texture, validating that the formulations met the expected requirements

Stability Test Results:

A two-week stability test confirmed that all sunscreen formulations containing Ficus racemosa L. ethanol extract remained physically stable, with no significant changes in color, odor, or texture. Each formulation (F0–F3) consistently maintained its characteristic appearance and semi-solid consistency, demonstrating excellent stability and supporting their suitability for further application and long-term effectiveness.

Irritation Test Results on Volunteers:

The irritation test was conducted to assess the potential for skin reactions caused by the sunscreen cream formulations. The results indicated that formulations F0 (blank), F1 (5%), F2 (7.5%), and F3 (10%) did not cause any visible signs of irritation, such as redness, itching, or swelling, when applied to the skin behind the ear of the volunteers. These findings suggest that the sunscreen cream formulations are safe for use and do not pose a risk of irritation upon application.

Results of Determining the SPF Value of Cream Preparations:

Table 7. SPF and Irritation Test Results for Volunteers

Formulation	Conc. (%)	Average SPF value	Category Protection	Irritation Result
Blank (F0)	0	1.60	Minimal	No irritation
F1	5	5.75	Moderate	No irritation
F2	7.5	7.24	Moderate	No irritation
F3	10	8.82	Maximum	No irritation

Legend:

SPF evaluation shows a concentration-dependent increase in photoprotective capacity, with F3 achieving maximum protection. The absence of irritation across all formulations confirms their safety for topical application, supporting both the efficacy and tolerability of the developed sunscreen creams.

The SPF evaluation showed that formulation 1 (5%) and formulation 2 (7.5%) demonstrated moderate protection, while formulation 3 (10%) achieved maximum protection. A clear dose-dependent relationship was observed, as higher extract concentrations yielded higher SPF values. SPF values were determined using the UV spectrophotometric method and classified according to COLIPA guidelines (The European Cosmetic, Toiletry and Perfumery Association), which define low protection (SPF 2–4), moderate protection (SPF 4–6), high protection (SPF 6–8), maximum protection (SPF 8–15), and ultra protection (SPF >15). Statistical analysis confirmed a significant increase in SPF with increasing extract concentration (p = 0.009), consistent with the dose-dependent photoprotective effect of flavonoid-rich extracts⁴⁰. Statistical evaluation revealed a significant increase in SPF values with higher extract concentrations (p = 0.009), consistent with the dose-dependent photoprotective effect of flavonoid-rich extracts.

CONCLUSION:

This study demonstrated that ethanol extract of Ficus racemosa L. obtained through microwave-assisted extraction (MAE) contains appreciable levels of flavonoids and exhibits strong antioxidant capacity, particularly in the ethyl acetate fraction. Sunscreen formulations incorporating the extract showed concentration-dependent SPF enhancement, with the 10% formulation achieving maximum protection, while maintaining stability, acceptable physicochemical characteristics, and safety in irritation testing.

The novelty of this work lies in the integration of MAE with solvent fractionation to optimize bioactive recovery, highlighting F. racemosa L. as a potent natural source of antioxidants and photoprotective agents. These findings provide a scientific basis for the development of safe, eco-friendly sunscreen formulations and open avenues for the broader application of plant-derived bioactives in cosmeceuticals.

ACKNOWLEDGMENTS:

The authors express gratitude for the facilities and scientific and technical assistance provided by Institut Kesehatan Helvetia, which supported this research through the provision of space and equipment.

CONFLICT OF INTERESTS:

The authors declare no conflict of interest.

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Received on 21.02.2025 Revised on 17.07.2025

Accepted on 04.10.2025 Published on 13.01.2026

Available online from January 17, 2026

Research J. Pharmacy and Technology. 2026;19(1):177-185.

DOI: 10.52711/0974-360X.2026.00027

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

Test Solution Concentration (ppm)	Measurement			% inhibition			Average % inhibition
Test Solution Concentration (ppm)	1	2	3	1	2	3	Average % inhibition
Blank	0.890	0.890	0.890	0	0	0	0
1	0.758	0.756	0.754	14.831	15.056	15.280	15.055
2	0.623	0.622	0.622	30.000	30.112	30.112	30.074
3	0.554	0.549	0.542	37.752	38.314	39.101	38.389
4	0.449	0.446	0.443	49.550	49.887	50.224	49.887
5	0.299	0.291	0.289	66.404	67.303	67.528	67.078

Sample	Stain Distance	Distance (cm)		Rf	Stain Color in 254 nm UV Lamp	positive compound
Sample	Stain Distance	Stain	Eluent	Rf	Stain Color in 254 nm UV Lamp	positive compound
Water Fraction	1	1.00	4.5	0.22	Black	Flavonoids
	2	2.00	4.5	0.44	Black	Flavonoids
	3	3.30	4.5	0.73	Black	Flavonoids
Ethyl Acetate Fraction	1	0.70	4.5	0.15	Black	-
	2	0.90	4.5	0.20	Black	Flavonoids
	3	2,30	4.5	0.51	Black	Flavonoids
	4	3.30	4.5	0.73	Black	Flavonoids
N-Hexane Fraction	-	-	-	-	-	-
Quercetin	1	3,20	4.5	0.71	Black	Flavonoids