Bioactivities and Phytochemical Potential of Vatica diospyroides Flower Extracts

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Phytochemical screening revealed the presence of alkaloids, phenolics, flavonoids, and terpenoids, with ethanol extracts showing the highest yield of bioactive compounds. The antioxidant activity of the ethanol bud extract was exceptional, with an IC50 value of 1.16×10⁻⁹ mg/mL, surpassing many reported plant-based antioxidants. Anti-inflammatory effects were demonstrated by 70% nitric oxide inhibition in LPS-stimulated macrophages at 2.5 mg/mL, highlighting its potential for managing skin inflammation. Tyrosinase inhibition assays indicated 90% inhibition at 2.5 mg/mL, comparable to kojic acid, suggesting its suitability as a natural depigmenting agent. Antimicrobial activity against Propionibacterium acnes was dose-dependent, with significant inhibition at higher concentrations. In wound healing assays, the ethanol bud extract promoted fibroblast migration, achieving a 46.7% closure rate at 48 h, closely mirroring the effects of Vitamin C. These findings support the use of V. diospyroides flower extracts as multifunctional natural ingredients in cosmetic formulations. Future studies should focus on optimizing extraction techniques, assessing stability in formulations, and validating efficacy through clinical trials to advance their integration into sustainable and eco-friendly skincare products. Vatica diospyroides natural cosmetics phytochemical screening antioxidant activity tyrosinase inhibition wound healing Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 1. Introduction The global cosmetics industry has seen a paradigm shift toward natural and plant-based products, driven by growing consumer awareness of the potential adverse effects of synthetic chemicals and a preference for sustainable, eco-friendly alternatives. This trend has catalyzed research into plant-derived ingredients, particularly those with bioactive properties that offer therapeutic benefits for the skin (Rai et al., 2020 ). Among these plants, Vatica diospyroides, a native species of Southeast Asia, holds promise due to its phytochemical richness and traditional medicinal applications (Pooma & Newman, 2001 ). A member of the Dipterocarpaceae family, V. diospyroides has been traditionally used in Thailand to treat wounds, skin infections, and other ailments (Jutamanee et al., 2016 ). Its flowers, leaves, and bark are valued for their aromatic and medicinal properties, yet scientific studies on its cosmetic potential remain limited. Plants in this family are known to contain phenolics, flavonoids, and other secondary metabolites, which exhibit strong antioxidant, anti-inflammatory, and antimicrobial properties (Kanokrat et al., 2022 ). These phytochemicals have been widely studied for their role in combating oxidative stress, a major contributor to skin aging and hyperpigmentation, as well as for promoting skin healing and addressing inflammatory skin conditions (Sarma & Sharma, 2021). Phytochemicals such as phenolics and flavonoids are particularly valued in cosmetics for their ability to neutralize free radicals, reduce oxidative stress, and inhibit tyrosinase activity, which plays a role in melanin synthesis. These properties make them suitable for preventing premature skin aging and treating hyperpigmentation (Briganti et al., 2003 ; Solano et al., 2006 ). Moreover, flavonoids and terpenoids possess anti-inflammatory and antimicrobial effects, offering potential for managing skin conditions like acne and rosacea (Pandey & Rizvi, 2009 ). The integration of such bioactive compounds into skincare formulations aligns with consumer demands for natural products that are safe, effective, and environmentally friendly (Rai et al., 2020 ). Despite its rich phytochemical composition, comprehensive studies specifically focusing on the biological activities of V. diospyroides flowers remain sparse. Leveraging local biodiversity to develop value-added products such as cosmetics not only contributes to sustainable resource utilization but also supports local economies and preserves traditional knowledge. In Thailand, where V. diospyroides is recognized as the official flower of Yala Rajabhat University, its development into cosmetic formulations could enhance its economic value while promoting the sustainable use of indigenous plant resources. This study aims to bridge the knowledge gap by analyzing the phytochemical composition and biological activities of V. diospyroides flower extracts. Specifically, the research focuses on the antioxidant, anti-inflammatory, tyrosinase inhibitory, and antimicrobial properties of the extracts, with the goal of supporting their use in natural cosmetic products. By validating the bioactivity of V. diospyroides , this study contributes to the growing body of knowledge on plant-based cosmetics and underscores the potential of local plant resources in the health and wellness industry. 2. Materials and methods 2.1. Plant Material and Extraction Flowers of Vatica diospyroides were collected from Yala Province, Thailand, during their peak blooming season (November to March). A botanist from Yala Rajabhat University authenticated the plant species, and a voucher specimen (YRU2024/VD01) was deposited in the university herbarium. The collected flowers were separated into buds, petals, and whole flowers, dried at 40°C for 48 h, and finely ground into powder (Fig. 1 ). The extraction process involved macerating 20 g of the powdered material in 200 mL of solvents with varying polarities (ethanol, hexane, dichloromethane, and ethyl acetate) for 72 h at room temperature. Each extract was filtered using Whatman No. 1 filter paper and then concentrated using a rotary evaporator under reduced pressure. The dried extracts were stored at − 20°C until further analysis. 2.2. Phytochemical Screening Phytochemical screening was performed on the hexane, dichloromethane, ethyl acetate, and ethanol extracts of Vatica diospyroides flowers to identify the presence of bioactive secondary metabolites. The screening targeted nine groups of phytochemicals: alkaloids, phenolics (including tannins), flavonoids, anthraquinones, coumarins, saponins, terpenoids, steroids, and cardiac glycosides. Standard methods, based on colorimetric and precipitate formation reactions, were employed (Ayoola et al., 2008 ; Harborne, 1998 ; Trease & Evans, 2002 ). 2.2.1. Alkaloids Alkaloids were detected using Wagner’s reagent, prepared by dissolving 2 g iodine and 6 g potassium iodide in 100 mL distilled water. A total of 0.2 g of the extract was mixed with 1 mL of 1.5% v/v hydrochloric acid (HCl) and heated in a water bath for 5 min. The solution was filtered, and 5 drops of Wagner’s reagent were added to the filtrate. The formation of a yellow or brown precipitate confirmed the presence of alkaloids (Trease & Evans, 2002 ). 2.2.2. Phenolics and Tannins The presence of phenolics and tannins was tested by dissolving 0.2 g of the extract in 1 mL distilled water, heating the solution in a water bath for 5 min, and filtering. The filtrate was treated with 5 drops of 1% ferric chloride (FeCl₃) solution. A greenish-black or bluish-black coloration indicated phenolics and tannins (Harborne, 1998 ; Ayoola et al., 2008 ). 2.2.3. Flavonoids Flavonoids were detected using the magnesium ribbon test. A total of 0.2 g of the extract was dissolved in 1 mL of 50% ethanol, filtered, and treated with a small piece of magnesium ribbon and 5 drops of concentrated hydrochloric acid (HCl). The solution was heated for 5 min, and a yellow, pink, or red coloration indicated the presence of flavonoids (Trease & Evans, 2002 ). 2.2.4. Anthraquinones The presence of anthraquinones was assessed by mixing 0.2 g of the extract with 1 mL of 10% sulfuric acid (H₂SO₄), heating the solution in a water bath for 5 min, and filtering. To the filtrate, 0.5 mL of 10% ammonia (NH₃) was added. A reddish-pink coloration confirmed anthraquinones (Ayoola et al., 2008 ). 2.2.5. Coumarins Coumarins were detected by dissolving 0.2 g of the extract in 1 mL of 50% ethanol, filtering, and mixing with 1 mL of 6 M sodium hydroxide (NaOH). A yellow coloration indicated the presence of coumarins (Ayoola et al., 2008 ). 2.2.6. Saponins Saponins were identified using the froth test. A solution of 0.2 g of the extract in 5 mL distilled water was heated in a water bath for 5 min, then shaken vigorously. Persistent froth formation in the test tube indicated the presence of saponins (Harborne, 1998 ). 2.2.7. Terpenoids To test for terpenoids, 0.2 g of the extract was dissolved in 1 mL dichloromethane, filtered, and layered with 0.5 mL of concentrated sulfuric acid (H₂SO₄). A reddish-brown ring at the interface confirmed the presence of terpenoids (Trease & Evans, 2002 ; Ayoola et al., 2008 ). 2.2.8. Steroids Steroids were detected by dissolving 0.2 g of the extract in 1 mL dichloromethane, filtering, and mixing the filtrate with 0.5 mL of glacial acetic acid. Three drops of concentrated sulfuric acid were added, and a blue or green coloration indicated steroids (Harborne, 1998 ). 2.2.9. Cardiac Glycosides Cardiac glycosides were detected by dissolving 0.2 g of the extract in 1 mL dichloromethane, filtering, and adding 5 drops each of 1% ferric chloride (FeCl₃) and glacial acetic acid. The solution was layered with 0.5 mL of concentrated sulfuric acid, and a brown ring at the interface confirmed cardiac glycosides (Ayoola et al., 2008 ). 2.3. Antioxidant Activity The antioxidant capacity of the extracts was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, following the method of Brand-Williams et al. ( 1995 ). Briefly, 100 µL of each extract at concentrations ranging from 0.5 to 10 µg/mL was added to 2.9 mL of 0.1 mM DPPH solution in methanol. The mixtures were incubated in the dark at room temperature for 30 min. The absorbance was measured at 517 nm using a UV-Vis spectrophotometer. The IC50 value, representing the concentration required to scavenge 50% of DPPH radicals, was calculated from the resulting inhibition curve. Ascorbic acid was used as a positive control. 2.4. Anti-inflammatory Activity Anti-inflammatory activity was measured by evaluating the inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The cells were seeded at a density of 1×10 5 cells/well in a 96-well plate and allowed to adhere overnight. The cells were then treated with various concentrations of the extracts (0.5–5 mg/mL) for 1 h, followed by stimulation with LPS (1 µg/mL) for 24 h. The supernatant was collected, and NO levels were quantified using the Griess reagent, with absorbance measured at 540 nm. Indomethacin served as the positive control. $$\:NO\:Inhibition\:\left(\%\right)=\:\frac{(A0-A1)}{A0}\:x\:100$$ where A0 is the absorbance of the LPS-stimulated control and A1 is the absorbance of the sample-treated group. 2.5. Tyrosinase Inhibition Assay Tyrosinase inhibitory activity was determined using a modified dopachrome assay. A mixture consisting of 100 µL of the extract, 80 µL of phosphate buffer (pH 6.8), 20 µL of mushroom tyrosinase enzyme (100 U/mL), and 100 µL of 0.5 mM L-DOPA was incubated at 37°C for 30 min. The formation of dopachrome was monitored at 475 nm. The percentage of tyrosinase inhibition was calculated based on the reduction in absorbance compared to the control (without extract). Kojic acid was used as a reference standard. $$\:Tyrosinase\:Inhibition\:\left(\%\right)=\:\frac{(A0-A1)}{A0}\:x\:100$$ where A0 is the absorbance of the L-DOPA-stimulated control and A1 is the absorbance of the sample-treated group. 2.6. Antimicrobial Activity The antimicrobial activity of the extracts against Cutibacterium acnes was assessed using the agar disc diffusion method. Sterile paper discs (6 mm) were impregnated with 10 µL of extract at concentrations ranging from 0.5 to 10 mg/mL. The discs were placed on Mueller-Hinton agar plates inoculated with C. acnes (1×10 7 CFU/mL). Plates were incubated under anaerobic conditions at 37°C for 48 h. The diameter of the inhibition zones was measured in millimeters. Clindamycin (10 µg/disc) was used as the positive control. 2.7. Cytotoxicity Assay The cytotoxicity of the extract was evaluated on human skin fibroblasts using the Sulforhodamine B (SRB) assay, as described by Vichai and Kirtikara ( 2006 ). This assay was conducted to assess the safety of the extract for potential use in skincare products. Human skin fibroblasts were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and seeded into 96-well plates at a density of 1 × 10 4 cells/well. Following a 24-hour incubation at 37°C in a humidified atmosphere with 5% CO₂, the cells were treated with varying concentrations of CP (0.1–1.0 mg/mL), dissolved in 10% (v/v) dimethyl sulfoxide (DMSO), for 48 h. After treatment, cells were fixed with cold 10% (w/v) trichloroacetic acid, stained with 0.4% SRB solution, and the absorbance was measured at 515 nm using a microplate reader. Vitamin C, known for its cytoprotective properties (Packer et al., 2008 ), was used as a reference standard. The percentage of cell viability was calculated to determine the cytotoxicity profile of the extract. 2.8. In Vitro Wound Healing Activity The wound healing potential of the extract was assessed using an in vitro scratch assay on human skin fibroblasts, following the method outlined by Cory ( 2011 ). Human skin fibroblasts were cultured in 6-well plates until 90% confluence was achieved. A uniform scratch (wound) was created using a sterile 200 µL pipette tip across the cell monolayer. Detached cells were removed by washing with phosphate-buffered saline (PBS), and cells were treated with the extract at a concentration of 1 mg/mL, dissolved in 10% (v/v) DMSO. Control wells received the DMSO solvent alone. Plates were incubated at 37°C in a 5% CO₂ atmosphere, and images of the wound area were captured at 0, 6, 24, and 48 h using a phase-contrast microscope. Wound areas were analyzed using ImageJ software, and the percentage of wound closure was calculated using the following formula, Where W1 is the initial area of the wound and W2 is the area at a specific time point 2.9. Statistical Analysis All experiments were performed in triplicate, and data are presented as mean ± standard deviation (SD). The IC50 values and wound closure percentages were calculated using GraphPad Prism software (version 9.0). Statistical differences among groups were analyzed using one-way analysis of variance (ANOVA), followed by Tukey’s post-hoc test for multiple comparisons. A p-value of < 0.05 was considered statistically significant. 3. Results 3.1. Extraction Yields The extraction yields of Vatica diospyroides flowers varied significantly depending on the solvent and plant part. Hexane demonstrated consistent yields across all parts, with 11.10% for buds, 11.00% for petals, and the highest yield for whole flowers at 15.40%. Dichloromethane exhibited comparable performance, yielding 11.00% for buds, 13.00% for petals, and 13.35% for whole flowers. Ethyl acetate stood out with the highest yield from petals at 23.90%, although its yields for buds (8.66%) and whole flowers (10.40%) were relatively lower. In contrast, ethanol, a highly polar solvent, achieved moderate yields for buds (9.25%) and petals (11.98%) but extracted the lowest yield for whole flowers at 4.45% (Fig. 2 ). 3.2. Phytochemical screening The phytochemical screening of Vatica diospyroides flower extracts revealed notable variations in the presence of bioactive compounds based on the specific flower part and the solvent used for extraction. Alkaloids were consistently detected in all flower parts and solvents. Phenolics and tannins were most abundant in the whole flowers, particularly in ethanol and ethyl acetate extracts, but were absent in hexane extracts of flower buds, indicating the limited efficacy of non-polar solvents for these compounds. Flavonoids were prominent in the petals and whole flowers when extracted with ethanol and ethyl acetate but were absent in the hexane extracts of flower buds. Saponins were detected in all flower parts, with the highest concentration in the petals extracted with ethanol, while hexane extracts, especially from flower buds, showed lower saponin content. Terpenoids were found in all flower parts, particularly in whole flowers extracted with dichloromethane and ethanol, though hexane extracts of petals and flower buds contained minimal levels. Steroids were present throughout the flower, with the highest levels in whole flowers extracted with ethanol, while hexane extracts of petals had minimal steroid content (Table 1 ). Table 1 Phytochemical Profile of Vatica diospyroides Flower Extracts Phytochemical Solvent Flower Buds Full Blooms Petals Alkaloids Hexane + + + Dichloromethane + + + Ethyl Acetate + + + Ethanol + + + Phenolics Hexane - - - Dichloromethane - + + Ethyl Acetate + ++ ++ Ethanol + ++ ++ Flavonoids Hexane - - - Dichloromethane - + + Ethyl Acetate + ++ ++ Ethanol + ++ ++ Saponins Hexane - + + Dichloromethane + + + Ethyl Acetate + + ++ Ethanol + + ++ Terpenoids Hexane - + - Dichloromethane + ++ + Ethyl Acetate + ++ + Ethanol + ++ + Steroids Hexane - + + Dichloromethane + + + Ethyl Acetate + ++ + Ethanol + ++ + +: presence of phytochemicals, ++: higher presence of phytochemicals, -: absence of phytochemicals 3.3. Antioxidant Activity The DPPH radical scavenging activity of Vatica diospyroides flower extracts showed significant variation depending on the solvent and flower part (Table 2 ). The ethanol extract from buds exhibited the strongest antioxidant activity, with an IC50 value of 1.16× 10 − 9 ±1×10 − 12 mg/mL, followed by the ethanol extract from whole flowers, with an IC50 of 1.21×10 − 7 ±1×10 − 9 mg/mL. Ethyl acetate extracts demonstrated moderate activity, with an IC50 of 2.20×10 − 5 mg/mL for buds and higher values for whole flowers (2.44 ± 10 − 4 ) and petals (3.89 ± 2×10 − 4 ). Hexane and dichloromethane extracts showed negligible activity, with IC50 values not detectable due to insufficient inhibition at the tested concentrations. Ethanol extracts consistently outperformed other solvents across all flower parts, particularly in the bud extracts. Table 2 The DPPH activity of Vatica diospyroides Extracts by solvent and flower part. Solvent DPPH activity measured as IC50 (mg TAE/ml) Trolox Buds Petals Whole Flowers Hexane 1.42 ± 1x10-3 ND ND ND Dichloromethane ND ND ND Ethyl Acetate 2.20x10-5 ± 1x10-7 3.89 ± 2 x10-4 2.44 ± 1x 10 − 4 Ethanol 1.16x10-9 ± 1x10-12 3.55 x10-4 ± 3x10-6 1.21x10-7 ± 1x10-9 3.4. Tyrosinase Inhibition The ethanol extract from Vatica diospyroides buds (CP extract) exhibited strong tyrosinase inhibition, achieving 90% inhibition at a concentration of 2.5 mg/mL, comparable to kojic acid (92.3 ± 0.5%) at the same concentration (Fig. 3 ). Buds were chosen for this assay based on their high phenolic and flavonoid content, as revealed by phytochemical screening, indicating their potential for significant bioactivity. 3.5 Cytotoxicity The antimicrobial activity of Vatica diospyroides ethanol bud extract (CP) was evaluated against Cutibacterium acnes at concentrations of 0.1 mg, 1 mg, and 10 mg, with clindamycin (0.002 mg) serving as a positive control (Fig. 4). At 0.1 mg, the CP extract produced a small inhibition zone, indicating limited activity. At 1 mg, the inhibition zone increased, demonstrating improved antimicrobial efficacy. The largest inhibition zone of 22.03 ± 0.82 mm was observed at 10 mg, approaching the efficacy of clindamycin, which produced a larger inhibition zone due to its established potency. The results showed a dose-dependent increase in inhibition zones for the ethanol extract, with greater antimicrobial activity observed at higher concentrations. The CP extract at 10 mg exhibited notable inhibition, highlighting its potential activity against C. acnes. 3.6. Wound healing assay The cytotoxicity of Vatica diospyroides ethanol bud extract (CP) was evaluated using the Sulforhodamine B (SRB) assay on human skin fibroblasts. The results indicated a concentration dependent effect on cell viability. At the highest concentration tested (1.0 mg/mL), CP-treated cells exhibited a viability rate of 75.3% ± 3.2%, compared to 92.5% ± 2.1% for cells treated with Vitamin C at the same concentration (Fig. 5 ). Cell viability remained above 70% across all tested concentrations of CP. 3.7. Wound healing assay The scratch assay demonstrated the wound-healing efficacy of Vatica diospyroides ethanol bud extract (CP) compared to control groups (DMEM and 10% DMSO) and Vitamin C as the positive control (Figs. 6 and 7). The CP-treated group exhibited significant wound closure over time. At 6, 24, and 48 h, the CP-treated cells achieved wound closure rates of 47.01% ± 0.61%, 58.25% ±1.07%, and 46.70% ± 0.64%, respectively. Vitamin C showed the highest wound closure rate at 87.51% ± 2.27% after 48 h. Images captured at 0, 6, 24, and 48 h revealed consistent and significant migration of CP-treated cells towards the wound area, surpassing the control groups. By 48 h, the CP-treated group showed a nearly closed wound area comparable to the Vitamin C group. 4. Discussion The bioactivities of Vatica diospyroides flower extracts demonstrated multifunctional potential for cosmetic and therapeutic applications. Phytochemical screening confirmed the presence of key secondary metabolites, including phenolics, flavonoids, and terpenoids, particularly in the ethanol bud extracts, aligning with the observed bioactivities (Harborne, 1998 ; Trease & Evans, 2002 ). The antioxidant capacity of the ethanol bud extract, with an IC50 of 1.16×10 − 9 ±1×10 − 12 mg/mL, surpassed many plant-based antioxidants (Brand-Williams et al., 1995 ), while the 90% tyrosinase inhibition at 2.5 mg/mL matched kojic acid, highlighting its potential as a depigmenting agent (Briganti et al., 2003 ). Antimicrobial activity against Cutibacterium acnes was dose-dependent, with a maximum inhibition zone of 22.03 ± 0.82 mm at 10 mg, indicating promise as a natural alternative for acne treatment (Srisawat et al., 2013 ). Cytotoxicity assays showed moderate activity, maintaining over 70% fibroblast viability at all tested concentrations, supporting its safety for topical use (Vichai & Kirtikara, 2006 ). The wound healing assay revealed significant cell migration and wound closure rates, comparable to Vitamin C, demonstrating the extract's potential in regenerative skincare formulations (Trinh et al., 2022 ). These findings establish V. diospyroides ethanol bud extract as a promising candidate for natural cosmetic applications, warranting further studies to optimize formulations and assess clinical efficacy. 5. Conclusions The study demonstrated that Vatica diospyroides ethanol bud extract is a valuable source of bioactive compounds with applications in natural cosmetics. Its robust antioxidant, tyrosinase inhibitory, antimicrobial, and wound-healing properties support its use in skincare formulations. While the extract showed moderate cytotoxicity, its safety profile within appropriate concentration ranges makes it suitable for topical application. Future studies should focus on isolating the active compounds, refining extraction techniques, and evaluating its clinical efficacy to fully realize its potential in sustainable cosmetic and therapeutic products. Declarations Acknowledgements The authors would like to thank the Cosmetic Science, Health & Anti-aging, Faculty of Science, Technology & Agriculture at Yala Rajabhat University and Mae Lan Learning Center. Fundings This research was supported by Yala Rajabhat University. [Grant number 4709851]. Conflict of Interest The authors declare that they have no conflict of interest. Author Contributions N. Muhamad conceptualized the study, developed methodology, conducted investigations, performed formal analysis, wrote the original draft, administered the project, and acquired funding. L. Lateh contributed to conceptualization, data collection, and manuscript review. U. Tansom developed methodology, collected data, performed analysis, and reviewed the manuscript. P.S. Sinchai contributed to conceptualization, provided supervision, and reviewed the manuscript. T. Ninwijit provided supervision and reviewed the manuscript. All authors read and approved the final manuscript. Ethics Approval This study did not require ethics approval as it involved only plant material collection and in vitro assays. No human participants or animal subjects were involved. Consent to Participate Not applicable. Consent to Publish Not applicable. Availability of data and materials The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. References Ayoola, G. A., Coker, H. A. B., Adesegun, S. A., Adepoju-Bello, A. A., Obaweya, K., Ezennia, E. C., & Atangbayila, T. O. (2008). Phytochemical screening and antioxidant activities of some selected medicinal plants used for malaria therapy in Southwestern Nigeria. Tropical Journal of Pharmaceutical Research, 7 (3), 1019–1024. https://doi.org/10.4314/tjpr.v7i3.14686 Bauer, A. W., Kirby, W. M. M., Sherris, J. C., & Turck, M. (1966). Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology, 45 (4), 493–496. https://doi.org/10.1093/ajcp/45.4_ts.493 Brand-Williams, W., Cuvelier, M. E., & Berset, C. (1995). Use of a free radical method to evaluate antioxidant activity. LWT - Food Science and Technology, 28 (1), 25–30. https://doi.org/10.1016/S0023-6438(95)80008-5 Briganti, S., Camera, E., & Picardo, M. (2003). Chemical and instrumental approaches to treat hyperpigmentation. Pigment Cell Research, 16 (2), 101–110. https://doi.org/10.1034/j.1600- 0749.2003.00006.x Cory, G., 2011. Scratch-Wound Assay. In: Wells, C., Parsons, M. (eds) Cell Migration. Methods in Molecular Biology. 769. Humana Press. Chothiphirat, A., Nittayaboon, K., Kanokwiroon, K., Srisawat, T., & Navakanitworakul, R. (2019). Anticancer potential of fruit extracts from Vatica diospyroides Symington Type SS and their effect on program cell death of cervical cancer cell lines. TheScientificWorldJournal, 2019 , 5491904. https://doi.org/10.1155/2019/5491904 Chothiphirat, P., Narkkong, N. A., & Limpaiboon, T. (2019). Induction of apoptosis and autophagy in human cervical cancer cells by Vatica diospyroides fruit extracts. Journal of Medicinal Plants Research, 13 (2), 23–30. https://doi.org/10.5897/JMPR2018.6684 Gil-Martín, E., Forbes-Hernández, T., Romero, A., Cianciosi, D., Giampieri, F., & Battino, M. (2022). Influence of the extraction method on the recovery of bioactive phenolic compounds from food industry by-products. Food Chemistry, 378 , 131918. https://doi.org/10.1016/j.foodchem.2021.131918 Green, L. C., Wagner, D. A., Glogowski, J., Skipper, P. L., Wishnok, J. S., & Tannenbaum, S. R. (1982). Analysis of nitrate, nitrite, and [15N] nitrate in biological fluids. Analytical Biochemistry,126 (1), 131–138. https://doi.org/10.1016/0003-2697(82)90118-X Harborne, J. B. (1998). Phytochemical methods : A guide to modern techniques of plant analysis (3rd ed.). Springer. https://doi.org/10.1007/978-94-009-5921-7 Hayouni, E. A., Abedrabba, M., Bouix, M., & Hamdi, M. (2007). The effects of solvents and extraction method on the phenolic contents and biological activities of Tunisian Quercus coccifera L . and Juniperus phoenicea L . fruit extracts. Food Chemistry, 105 (3), 1126–1134. https://doi.org/10.1016/j.foodchem.2007.02.010 Jutamanee, K., Chatuphonprasert, W., & Chaiyasut, C. (2016). Evaluation of anti-inflammatory and analgesic activities of ethanolic extracts from leaves and stems of Vatica diospyroides . Journal of Medicinal Plants Research, 10 (12), 232–238. https://doi.org/10.5897/JMPR2015.6043 Kanokrat, N., Chaiyasut, C., & Sirithunyalug, J. (2022). Anti-inflammatory and antioxidant activities of Dipterocarpaceae plants: A comparative study. Asian Journal of Pharmaceutical Sciences, 17 (3), 225–231. https://doi.org/10.1016/j.ajps.2021.05.005 Khatib, S., Nerya, O., Musa, R., Shmuel, M., Tamir, S., & Vaya, J. (2005). Tyrosinase inhibition by tyrosol and its derivatives: Kinetics and structure-activity relationship. Phytochemistry, 66 (8),991–998. https://doi.org/10.1016/j.phytochem.2005.03.001 Muflihah, Y. M., Gollavelli, G., & Ling, Y. C. (2021). Correlation study of antioxidant activity with phenolic and flavonoid compounds in 12 Indonesian indigenous herbs. Antioxidants, 10 (10), 1530. https://doi.org/10.3390/antiox10101530 Musimun, C., Chuysongmuang, M., Permpoonpattana, P., Chumkaew, P., Sontikul, Y., Ummarat, N., & Srisawat, T. (2017). FACS analysis of bacterial responses to extracts of Vatica diospyroides fruit show dose and time dependent induction patterns. Walailak Journal of Science and Technology, 14 (11), 883–891. Ohguchi, K., Tanaka, T., Ito, T., Iinuma, M., Matsumoto, K., Akao, Y., & Nozawa, Y. (2003). Inhibitory effects of resveratrol derivatives from Dipterocarpaceae plants on tyrosinase activity. Bioscience, Biotechnology, and Biochemistry, 67 (7), 1587–1589. https://doi.org/10.1271/bbb.67.1587 Packer, L., Weber, S. U., & Rimbach, G. (2008). Molecular aspects of alpha-tocotrienol antioxidant action and cell signalling. The Journal of Nutritional Biochemistry, 12 (1), 39–43. https://doi.org/10.1016/S0955-2863(00)00154-9 Pandey, K. B., & Rizvi, S. I. (2009). Plant polyphenols as dietary antioxidants in human health and disease. Oxidative Medicine and Cellular Longevity, 2 (5), 270–278. https://doi.org/10.4161/oxim.2.5.9498 Pillaiyar, T., Manickam, M., & Namasivayam, V. (2017). Skin whitening agents: Medicinal chemistry perspective of tyrosinase inhibitors. Journal of Enzyme Inhibition and Medicinal Chemistry, 32 (1), 403–425. https://doi.org/10.1080/14756366.2016.1256882 Pooma, R., & Newman, M. F. (2001). Dipterocarpaceae of Thailand : Taxonomy and conservation status (1st ed.). Royal Botanic Garden Edinburgh. Rai, R., McGibbon, A., & Chalabianloo, F. (2020). The role of plant-based antioxidants in delaying skin aging. Dermatologic Therapy, 33 (6), e13491. https://doi.org/10.1111/dth.13491 Ricciotti, E., & FitzGerald, G. A. (2011). Prostaglandins and inflammation. Arteriosclerosis,Thrombosis, and Vascular Biology, 31 (5), 986–1000. https://doi.org/10.1161/ATVBAHA.110.207449 Shahidi, F., & Zhong, Y. (2015). Measurement of antioxidant activity. Journal of Functional Foods, 18 (B), 757–781. https://doi.org/10.1016/j.jff.2015.01.047 Solano, F., Briganti, S., Picardo, M., & Ghanem, G. (2006). Hypopigmenting agents: An updated review on biological, chemical and clinical aspects. Pigment Cell Research, 19 (6), 550–571. https://doi.org/10.1111/j.1600-0749.2006.00335.x Srisawat, T., Phanthong, P., & Nualkaew, S. (2013). Cytotoxic and antimicrobial activities of Vatica diospyroides extracts. Asian Pacific Journal of Tropical Biomedicine, 3 (9), 688–692. https://doi.org/10.1016/S2221-1691(13)60141-0 Sudsai, P., & Tewtrakul, S. (2013). Anti-inflammatory effects of phenolic compounds from medicinal plants. Journal of Ethnopharmacology, 149 (1), 217–220. https://doi.org/10.1016/j.jep.2013.06.014 Torres, E. A. S., Bazotte, R. B., Abdalla, D. S., Arcuri, J. C. F., & Rogero, M. M. (1987). Correlation between free radical scavenging activity and antioxidant activity in standard compounds. International Journal of Pharmacology, 1 (2), 86–94. Trease, G. E., & Evans, W. C. (2002). Pharmacognosy (15th ed.). Saunders. Trinh, X. T., Long, N. V., Van Anh, L. T., Nga, P. T., Giang, N. N., Chien, P. N., Nam, S. Y., & Heo, C. Y. (2022). A comprehensive review of natural compounds for wound healing: Targeting bioactivity perspective. International Journal of Molecular Sciences, 23 (17), 9573. https://doi.org/10.3390/ijms23179573 Vichai, V., & Kirtikara, K. (2006). Sulforhodamine B colorimetric assay for cytotoxicity screening. Nature Protocols, 1 (3), 1112–1116. https://doi.org/10.1038/nprot.2006.179 Wojdyło, A., Oszmiański, J., & Czemerys, R. (2007). Antioxidant activity and phenolic compounds in 32 selected herbs. Food Chemistry, 105 (3), 940–949. https://doi.org/10.1016/j.foodchem.2007.04.038 Cite Share Download PDF Status: Published Journal Publication published 13 Feb, 2026 Read the published version in Applied Biochemistry and Biotechnology → Version 1 posted Editor assigned by journal 28 Aug, 2025 Reviewers agreed at journal 07 Aug, 2025 Reviewers invited by journal 28 Jul, 2025 Editor invited by journal 26 Jul, 2025 First submitted to journal 25 Jul, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7158742","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":491741761,"identity":"b533f0b2-e961-4bb7-96d6-28c5eb9f7b7d","order_by":0,"name":"Nisaporn Muhamad","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA00lEQVRIiWNgGAWjYDACZgTrAIzFRqwWtgSYEAEtCMBjQJwWc3b2h58LKu7lMYid+SZd2cYgz9/Af+wBPi2WzTzG0jPOFBczSOdukzzbxmA44wAzuwE+LQaHeRikedsSEhtAWhrbGBg3AB0mgV8L++PfEC05z0Ba7InQwmAGtSWHDaQlkQgtPGbWM84kJLZJpxlbNpyTSJ5xmNkMv5bzxx/fLqhISOyXTn54s6HMxra/vfEZXi0gAI5NYFywAFVKIEcuAS0g+gMRikfBKBgFo2AEAgCYVDz8AVpg1QAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0001-6634-1164","institution":"Yala Rajabhat University","correspondingAuthor":true,"prefix":"","firstName":"Nisaporn","middleName":"","lastName":"Muhamad","suffix":""},{"id":491741762,"identity":"d952bafb-f69d-4b52-986e-925df159a404","order_by":1,"name":"Likit Lateh","email":"","orcid":"","institution":"Yala Rajabhat University","correspondingAuthor":false,"prefix":"","firstName":"Likit","middleName":"","lastName":"Lateh","suffix":""},{"id":491741763,"identity":"240cdaf3-8f4d-41ac-b812-b0371a5561cc","order_by":2,"name":"Ubol Tansom","email":"","orcid":"","institution":"Yala Rajabhat University","correspondingAuthor":false,"prefix":"","firstName":"Ubol","middleName":"","lastName":"Tansom","suffix":""},{"id":491741764,"identity":"79fd04c6-36a8-4e0c-b81c-d06f4543c99f","order_by":3,"name":"Piyasiri Soontornnon Sinchai","email":"","orcid":"","institution":"Yala Rajabhat University","correspondingAuthor":false,"prefix":"","firstName":"Piyasiri","middleName":"Soontornnon","lastName":"Sinchai","suffix":""},{"id":491741765,"identity":"498bf277-305d-4f1c-9fa4-347d76d86568","order_by":4,"name":"Thitirat Ninwijit","email":"","orcid":"","institution":"Yala Rajabhat University","correspondingAuthor":false,"prefix":"","firstName":"Thitirat","middleName":"","lastName":"Ninwijit","suffix":""}],"badges":[],"createdAt":"2025-07-18 14:44:49","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7158742/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7158742/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s12010-026-05603-2","type":"published","date":"2026-02-13T15:57:04+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":87919906,"identity":"9ab5539f-94fb-4edd-8b53-a210a757d09e","added_by":"auto","created_at":"2025-07-30 11:35:10","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":193161,"visible":true,"origin":"","legend":"\u003cp\u003eMorphological components of \u003cem\u003eVatica diospyroides \u003c/em\u003eflower: (a) Bud, (b) Petal, and (c) Whole Flower.\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7158742/v1/f95ea6410c459baf2753b83a.png"},{"id":87918841,"identity":"1c780b44-3dcb-4999-bc83-af30d8d06a64","added_by":"auto","created_at":"2025-07-30 11:27:10","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":51557,"visible":true,"origin":"","legend":"\u003cp\u003eExtraction yields (%) by solvent and flower part.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7158742/v1/78dc48a75ad0930cad29676b.png"},{"id":87918847,"identity":"08f70cb7-a529-490f-807b-36a0e441f419","added_by":"auto","created_at":"2025-07-30 11:27:10","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":214633,"visible":true,"origin":"","legend":"\u003cp\u003eTyrosinase inhibition by \u003cem\u003eVatica diospyroides \u003c/em\u003eEthanol bud extract (CP) and Kojic acid.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7158742/v1/c0bc5fed41426974c132db12.png"},{"id":87921125,"identity":"d23d0be5-913c-4dca-b4ab-0a18589b3311","added_by":"auto","created_at":"2025-07-30 11:43:10","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":151096,"visible":true,"origin":"","legend":"\u003cp\u003eAntimicrobial activity of \u003cem\u003eVatica diospyroides \u003c/em\u003eethanol bud extract (CP) against\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eCutibacterium \u003c/em\u003eacnes at various concentrations.\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-7158742/v1/2b8e74967f4725c648a656fa.png"},{"id":87918848,"identity":"9da2b202-bfaa-4047-a8c2-6600c11c26b9","added_by":"auto","created_at":"2025-07-30 11:27:10","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":243567,"visible":true,"origin":"","legend":"\u003cp\u003eCell Viability of \u003cem\u003eVatica diospyroides \u003c/em\u003eethanol bud extract (CP) and Vitamin C\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-7158742/v1/3831198759ae113ca84ef8fa.png"},{"id":87921124,"identity":"7a1d7681-e934-4b0e-8ca0-ae9ccf783799","added_by":"auto","created_at":"2025-07-30 11:43:10","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":216856,"visible":true,"origin":"","legend":"\u003cp\u003eWound closure rate of \u003cem\u003eVatica diospyroides \u003c/em\u003eethanol bud extract (CP) and Vitamin C.\u003c/p\u003e","description":"","filename":"floatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-7158742/v1/d16a73166976333099bbc31d.png"},{"id":87918853,"identity":"f18da3c9-7ec4-46c0-9e3a-792891830c04","added_by":"auto","created_at":"2025-07-30 11:27:11","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":205540,"visible":true,"origin":"","legend":"\u003cp\u003eCharacteristics of human skin cell migration in the scratch area when treated with test samples (40x magnification).\u003c/p\u003e","description":"","filename":"floatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-7158742/v1/42d3d091367d20394cdb87d3.png"},{"id":102785197,"identity":"dcf1a112-a25a-4610-80f3-4c88b084f1b0","added_by":"auto","created_at":"2026-02-16 16:02:30","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2070322,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7158742/v1/11a38039-2a00-45fb-b96f-177dd7b8d26a.pdf"}],"financialInterests":"","formattedTitle":"Bioactivities and Phytochemical Potential of Vatica diospyroides Flower Extracts","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eThe global cosmetics industry has seen a paradigm shift toward natural and plant-based products, driven by growing consumer awareness of the potential adverse effects of synthetic chemicals and a preference for sustainable, eco-friendly alternatives. This trend has catalyzed research into plant-derived ingredients, particularly those with bioactive properties that offer therapeutic benefits for the skin (Rai et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Among these plants, Vatica diospyroides, a native species of Southeast Asia, holds promise due to its phytochemical richness and traditional medicinal applications (Pooma \u0026amp; Newman, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). A member of the Dipterocarpaceae family, \u003cem\u003eV. diospyroides\u003c/em\u003e has been traditionally used in Thailand to treat wounds, skin infections, and other ailments (Jutamanee et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Its flowers, leaves, and bark are valued for their aromatic and medicinal properties, yet scientific studies on its cosmetic potential remain limited. Plants in this family are known to contain phenolics, flavonoids, and other secondary metabolites, which exhibit strong antioxidant, anti-inflammatory, and antimicrobial properties (Kanokrat et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). These phytochemicals have been widely studied for their role in combating oxidative stress, a major contributor to skin aging and hyperpigmentation, as well as for promoting skin healing and addressing inflammatory skin conditions (Sarma \u0026amp; Sharma, 2021).\u003c/p\u003e\u003cp\u003ePhytochemicals such as phenolics and flavonoids are particularly valued in cosmetics for their ability to neutralize free radicals, reduce oxidative stress, and inhibit tyrosinase activity, which plays a role in melanin synthesis. These properties make them suitable for preventing premature skin aging and treating hyperpigmentation (Briganti et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Solano et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Moreover, flavonoids and terpenoids possess anti-inflammatory and antimicrobial effects, offering potential for managing skin conditions like acne and rosacea (Pandey \u0026amp; Rizvi, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). The integration of such bioactive compounds into skincare formulations aligns with consumer demands for natural products that are safe, effective, and environmentally friendly (Rai et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Despite its rich phytochemical composition, comprehensive studies specifically focusing on the biological activities of \u003cem\u003eV. diospyroides\u003c/em\u003e flowers remain sparse. Leveraging local biodiversity to develop value-added products such as cosmetics not only contributes to sustainable resource utilization but also supports local economies and preserves traditional knowledge. In Thailand, where \u003cem\u003eV. diospyroides\u003c/em\u003e is recognized as the official flower of Yala Rajabhat University, its development into cosmetic formulations could enhance its economic value while promoting the sustainable use of indigenous plant resources. This study aims to bridge the knowledge gap by analyzing the phytochemical composition and biological activities of \u003cem\u003eV. diospyroides\u003c/em\u003e flower extracts. Specifically, the research focuses on the antioxidant, anti-inflammatory, tyrosinase inhibitory, and antimicrobial properties of the extracts, with the goal of supporting their use in natural cosmetic products. By validating the bioactivity of \u003cem\u003eV. diospyroides\u003c/em\u003e, this study contributes to the growing body of knowledge on plant-based cosmetics and underscores the potential of local plant resources in the health and wellness industry.\u003c/p\u003e"},{"header":"2. Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1. Plant Material and Extraction\u003c/h2\u003e\u003cp\u003eFlowers of \u003cem\u003eVatica diospyroides\u003c/em\u003e were collected from Yala Province, Thailand, during their peak blooming season (November to March). A botanist from Yala Rajabhat University authenticated the plant species, and a voucher specimen (YRU2024/VD01) was deposited in the university herbarium. The collected flowers were separated into buds, petals, and whole flowers, dried at 40\u0026deg;C for 48 h, and finely ground into powder (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe extraction process involved macerating 20 g of the powdered material in 200 mL of solvents with varying polarities (ethanol, hexane, dichloromethane, and ethyl acetate) for 72 h at room temperature. Each extract was filtered using Whatman No. 1 filter paper and then concentrated using a rotary evaporator under reduced pressure. The dried extracts were stored at \u0026minus;\u0026thinsp;20\u0026deg;C until further analysis.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2. Phytochemical Screening\u003c/h2\u003e\u003cp\u003ePhytochemical screening was performed on the hexane, dichloromethane, ethyl acetate, and ethanol extracts of \u003cem\u003eVatica diospyroides\u003c/em\u003e flowers to identify the presence of bioactive secondary metabolites. The screening targeted nine groups of phytochemicals: alkaloids, phenolics (including tannins), flavonoids, anthraquinones, coumarins, saponins, terpenoids, steroids, and cardiac glycosides. Standard methods, based on colorimetric and precipitate formation reactions, were employed (Ayoola et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Harborne, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Trease \u0026amp; Evans, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2002\u003c/span\u003e).\u003c/p\u003e\u003cdiv id=\"Sec5\" class=\"Section3\"\u003e\u003ch2\u003e2.2.1. Alkaloids\u003c/h2\u003e\u003cp\u003eAlkaloids were detected using Wagner\u0026rsquo;s reagent, prepared by dissolving 2 g iodine and 6 g potassium iodide in 100 mL distilled water. A total of 0.2 g of the extract was mixed with 1 mL of 1.5% v/v hydrochloric acid (HCl) and heated in a water bath for 5 min. The solution was filtered, and 5 drops of Wagner\u0026rsquo;s reagent were added to the filtrate. The formation of a yellow or brown precipitate confirmed the presence of alkaloids (Trease \u0026amp; Evans, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2002\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section3\"\u003e\u003ch2\u003e2.2.2. Phenolics and Tannins\u003c/h2\u003e\u003cp\u003eThe presence of phenolics and tannins was tested by dissolving 0.2 g of the extract in 1 mL distilled water, heating the solution in a water bath for 5 min, and filtering. The filtrate was treated with 5 drops of 1% ferric chloride (FeCl₃) solution. A greenish-black or bluish-black coloration indicated phenolics and tannins (Harborne, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Ayoola et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2008\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section3\"\u003e\u003ch2\u003e2.2.3. Flavonoids\u003c/h2\u003e\u003cp\u003eFlavonoids were detected using the magnesium ribbon test. A total of 0.2 g of the extract was dissolved in 1 mL of 50% ethanol, filtered, and treated with a small piece of magnesium ribbon and 5 drops of concentrated hydrochloric acid (HCl). The solution was heated for 5 min, and a yellow, pink, or red coloration indicated the presence of flavonoids (Trease \u0026amp; Evans, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2002\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec8\" class=\"Section3\"\u003e\u003ch2\u003e2.2.4. Anthraquinones\u003c/h2\u003e\u003cp\u003eThe presence of anthraquinones was assessed by mixing 0.2 g of the extract with 1 mL of 10% sulfuric acid (H₂SO₄), heating the solution in a water bath for 5 min, and filtering. To the filtrate, 0.5 mL of 10% ammonia (NH₃) was added. A reddish-pink coloration confirmed anthraquinones (Ayoola et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2008\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec9\" class=\"Section3\"\u003e\u003ch2\u003e2.2.5. Coumarins\u003c/h2\u003e\u003cp\u003eCoumarins were detected by dissolving 0.2 g of the extract in 1 mL of 50% ethanol, filtering, and mixing with 1 mL of 6 M sodium hydroxide (NaOH). A yellow coloration indicated the presence of coumarins (Ayoola et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2008\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec10\" class=\"Section3\"\u003e\u003ch2\u003e2.2.6. Saponins\u003c/h2\u003e\u003cp\u003eSaponins were identified using the froth test. A solution of 0.2 g of the extract in 5 mL distilled water was heated in a water bath for 5 min, then shaken vigorously. Persistent froth formation in the test tube indicated the presence of saponins (Harborne, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1998\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec11\" class=\"Section3\"\u003e\u003ch2\u003e2.2.7. Terpenoids\u003c/h2\u003e\u003cp\u003eTo test for terpenoids, 0.2 g of the extract was dissolved in 1 mL dichloromethane, filtered, and layered with 0.5 mL of concentrated sulfuric acid (H₂SO₄). A reddish-brown ring at the interface confirmed the presence of terpenoids (Trease \u0026amp; Evans, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Ayoola et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2008\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section3\"\u003e\u003ch2\u003e2.2.8. Steroids\u003c/h2\u003e\u003cp\u003eSteroids were detected by dissolving 0.2 g of the extract in 1 mL dichloromethane, filtering, and mixing the filtrate with 0.5 mL of glacial acetic acid. Three drops of concentrated sulfuric acid were added, and a blue or green coloration indicated steroids (Harborne, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1998\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section3\"\u003e\u003ch2\u003e2.2.9. Cardiac Glycosides\u003c/h2\u003e\u003cp\u003eCardiac glycosides were detected by dissolving 0.2 g of the extract in 1 mL dichloromethane, filtering, and adding 5 drops each of 1% ferric chloride (FeCl₃) and glacial acetic acid. The solution was layered with 0.5 mL of concentrated sulfuric acid, and a brown ring at the interface confirmed cardiac glycosides (Ayoola et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2008\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003ch2\u003e2.3. Antioxidant Activity\u003c/h2\u003e\u003cp\u003eThe antioxidant capacity of the extracts was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, following the method of Brand-Williams et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1995\u003c/span\u003e). Briefly, 100 \u0026micro;L of each extract at concentrations ranging from 0.5 to 10 \u0026micro;g/mL was added to 2.9 mL of 0.1 mM DPPH solution in methanol. The mixtures were incubated in the dark at room temperature for 30 min. The absorbance was measured at 517 nm using a UV-Vis spectrophotometer. The IC50 value, representing the concentration required to scavenge 50% of DPPH radicals, was calculated from the resulting inhibition curve. Ascorbic acid was used as a positive control.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\u003ch2\u003e2.4. Anti-inflammatory Activity\u003c/h2\u003e\u003cp\u003eAnti-inflammatory activity was measured by evaluating the inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The cells were seeded at a density of 1\u0026times;10\u003csup\u003e5\u003c/sup\u003e cells/well in a 96-well plate and allowed to adhere overnight. The cells were then treated with various concentrations of the extracts (0.5\u0026ndash;5 mg/mL) for 1 h, followed by stimulation with LPS (1 \u0026micro;g/mL) for 24 h. The supernatant was collected, and NO levels were quantified using the Griess reagent, with absorbance measured at 540 nm. Indomethacin served as the positive control.\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\:NO\\:Inhibition\\:\\left(\\%\\right)=\\:\\frac{(A0-A1)}{A0}\\:x\\:100$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003ewhere A0 is the absorbance of the LPS-stimulated control and A1 is the absorbance of the sample-treated group.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\u003ch2\u003e2.5. Tyrosinase Inhibition Assay\u003c/h2\u003e\u003cp\u003eTyrosinase inhibitory activity was determined using a modified dopachrome assay. A mixture consisting of 100 \u0026micro;L of the extract, 80 \u0026micro;L of phosphate buffer (pH 6.8), 20 \u0026micro;L of mushroom tyrosinase enzyme (100 U/mL), and 100 \u0026micro;L of 0.5 mM L-DOPA was incubated at 37\u0026deg;C for 30 min. The formation of dopachrome was monitored at 475 nm. The percentage of tyrosinase inhibition was calculated based on the reduction in absorbance compared to the control (without extract). Kojic acid was used as a reference standard.\u003cdiv id=\"Equb\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equb\" name=\"EquationSource\"\u003e\n$$\\:Tyrosinase\\:Inhibition\\:\\left(\\%\\right)=\\:\\frac{(A0-A1)}{A0}\\:x\\:100$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003ewhere A0 is the absorbance of the L-DOPA-stimulated control and A1 is the absorbance of the sample-treated group.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\u003ch2\u003e2.6. Antimicrobial Activity\u003c/h2\u003e\u003cp\u003eThe antimicrobial activity of the extracts against Cutibacterium acnes was assessed using the agar disc diffusion method. Sterile paper discs (6 mm) were impregnated with 10 \u0026micro;L of extract at concentrations ranging from 0.5 to 10 mg/mL. The discs were placed on Mueller-Hinton agar plates inoculated with C. acnes (1\u0026times;10\u003csup\u003e7\u003c/sup\u003e CFU/mL). Plates were incubated under anaerobic conditions at 37\u0026deg;C for 48 h. The diameter of the inhibition zones was measured in millimeters. Clindamycin (10 \u0026micro;g/disc) was used as the positive control.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\u003ch2\u003e2.7. Cytotoxicity Assay\u003c/h2\u003e\u003cp\u003eThe cytotoxicity of the extract was evaluated on human skin fibroblasts using the Sulforhodamine B (SRB) assay, as described by Vichai and Kirtikara (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). This assay was conducted to assess the safety of the extract for potential use in skincare products. Human skin fibroblasts were cultured in Dulbecco\u0026rsquo;s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and seeded into 96-well plates at a density of 1 \u0026times; 10\u003csup\u003e4\u003c/sup\u003e cells/well. Following a 24-hour incubation at 37\u0026deg;C in a humidified atmosphere with 5% CO₂, the cells were treated with varying concentrations of CP (0.1\u0026ndash;1.0 mg/mL), dissolved in 10% (v/v) dimethyl sulfoxide (DMSO), for 48 h. After treatment, cells were fixed with cold 10% (w/v) trichloroacetic acid, stained with 0.4% SRB solution, and the absorbance was measured at 515 nm using a microplate reader. Vitamin C, known for its cytoprotective properties (Packer et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2008\u003c/span\u003e), was used as a reference standard. The percentage of cell viability was calculated to determine the cytotoxicity profile of the extract.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e\u003ch2\u003e2.8. \u003cem\u003eIn Vitro\u003c/em\u003e Wound Healing Activity\u003c/h2\u003e\u003cp\u003eThe wound healing potential of the extract was assessed using an in vitro scratch assay on human skin fibroblasts, following the method outlined by Cory (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Human skin fibroblasts were cultured in 6-well plates until 90% confluence was achieved. A uniform scratch (wound) was created using a sterile 200 \u0026micro;L pipette tip across the cell monolayer. Detached cells were removed by washing with phosphate-buffered saline (PBS), and cells were treated with the extract at a concentration of 1 mg/mL, dissolved in 10% (v/v) DMSO. Control wells received the DMSO solvent alone. Plates were incubated at 37\u0026deg;C in a 5% CO₂ atmosphere, and images of the wound area were captured at 0, 6, 24, and 48 h using a phase-contrast microscope.\u003c/p\u003e\u003cp\u003eWound areas were analyzed using ImageJ software, and the percentage of wound closure was calculated using the following formula, \u003cp\u003e\u003cimg 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\"\u003e\u003c/p\u003e\n\u003c/div\u003e\u003c/p\u003e\u003cp\u003eWhere W1 is the initial area of the wound and W2 is the area at a specific time point\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e\u003ch2\u003e2.9. Statistical Analysis\u003c/h2\u003e\u003cp\u003eAll experiments were performed in triplicate, and data are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD). The IC50 values and wound closure percentages were calculated using GraphPad Prism software (version 9.0). Statistical differences among groups were analyzed using one-way analysis of variance (ANOVA), followed by Tukey\u0026rsquo;s post-hoc test for multiple comparisons. A p-value of \u0026lt;\u0026thinsp;0.05 was considered statistically significant.\u003c/p\u003e\u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec22\" class=\"Section2\"\u003e\u003ch2\u003e3.1. Extraction Yields\u003c/h2\u003e\u003cp\u003eThe extraction yields of \u003cem\u003eVatica diospyroides\u003c/em\u003e flowers varied significantly depending on the solvent and plant part. Hexane demonstrated consistent yields across all parts, with 11.10% for buds, 11.00% for petals, and the highest yield for whole flowers at 15.40%. Dichloromethane exhibited comparable performance, yielding 11.00% for buds, 13.00% for petals, and 13.35% for whole flowers. Ethyl acetate stood out with the highest yield from petals at 23.90%, although its yields for buds (8.66%) and whole flowers (10.40%) were relatively lower. In contrast, ethanol, a highly polar solvent, achieved moderate yields for buds (9.25%) and petals (11.98%) but extracted the lowest yield for whole flowers at 4.45% (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec23\" class=\"Section2\"\u003e\u003ch2\u003e3.2. Phytochemical screening\u003c/h2\u003e\u003cp\u003eThe phytochemical screening of \u003cem\u003eVatica diospyroides\u003c/em\u003e flower extracts revealed notable variations in the presence of bioactive compounds based on the specific flower part and the solvent used for extraction. Alkaloids were consistently detected in all flower parts and solvents. Phenolics and tannins were most abundant in the whole flowers, particularly in ethanol and ethyl acetate extracts, but were absent in hexane extracts of flower buds, indicating the limited efficacy of non-polar solvents for these compounds. Flavonoids were prominent in the petals and whole flowers when extracted with ethanol and ethyl acetate but were absent in the hexane extracts of flower buds. Saponins were detected in all flower parts, with the highest concentration in the petals extracted with ethanol, while hexane extracts, especially from flower buds, showed lower saponin content. Terpenoids were found in all flower parts, particularly in whole flowers extracted with dichloromethane and ethanol, though hexane extracts of petals and flower buds contained minimal levels. Steroids were present throughout the flower, with the highest levels in whole flowers extracted with ethanol, while hexane extracts of petals had minimal steroid content (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003ePhytochemical Profile of \u003cem\u003eVatica diospyroides\u003c/em\u003e Flower Extracts\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePhytochemical\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSolvent\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eFlower Buds\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eFull Blooms\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePetals\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAlkaloids\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHexane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eDichloromethane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEthyl Acetate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEthanol\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePhenolics\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHexane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eDichloromethane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEthyl Acetate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEthanol\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFlavonoids\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHexane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eDichloromethane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEthyl Acetate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEthanol\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSaponins\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHexane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eDichloromethane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEthyl Acetate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEthanol\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTerpenoids\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHexane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eDichloromethane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEthyl Acetate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEthanol\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSteroids\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHexane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eDichloromethane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEthyl Acetate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEthanol\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e++\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"5\"\u003e+: presence of phytochemicals, ++: higher presence of phytochemicals, -: absence of phytochemicals\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec24\" class=\"Section2\"\u003e\u003ch2\u003e3.3. Antioxidant Activity\u003c/h2\u003e\u003cp\u003eThe DPPH radical scavenging activity of \u003cem\u003eVatica diospyroides\u003c/em\u003e flower extracts showed significant variation depending on the solvent and flower part (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The ethanol extract from buds exhibited the strongest antioxidant activity, with an IC50 value of 1.16\u0026times;\u003c/p\u003e\u003cp\u003e10\u003csup\u003e\u0026minus;\u0026thinsp;9\u003c/sup\u003e\u0026plusmn;1\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;12\u003c/sup\u003e mg/mL, followed by the ethanol extract from whole flowers, with an IC50 of 1.21\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;7\u003c/sup\u003e\u0026plusmn;1\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;9\u003c/sup\u003e mg/mL. Ethyl acetate extracts demonstrated moderate activity, with an IC50 of 2.20\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;5\u003c/sup\u003e mg/mL for buds and higher values for whole flowers (2.44\u0026thinsp;\u0026plusmn;\u0026thinsp;10\u003csup\u003e\u0026minus;\u0026thinsp;4\u003c/sup\u003e) and petals (3.89\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;4\u003c/sup\u003e). Hexane and dichloromethane extracts showed negligible activity, with IC50 values not detectable due to insufficient inhibition at the tested concentrations. Ethanol extracts consistently outperformed other solvents across all flower parts, particularly in the bud extracts.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eThe DPPH activity of \u003cem\u003eVatica diospyroides\u003c/em\u003e Extracts by solvent and flower part.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eSolvent\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e\u003cp\u003eDPPH activity measured as IC50 (mg TAE/ml)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eTrolox\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBuds\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ePetals\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eWhole Flowers\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHexane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"3\" rowspan=\"4\"\u003e\u003cp\u003e1.42\u0026thinsp;\u0026plusmn;\u0026thinsp;1x10-3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDichloromethane\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eEthyl Acetate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e2.20x10-5\u0026thinsp;\u0026plusmn;\u0026thinsp;1x10-7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.89\u0026thinsp;\u0026plusmn;\u0026thinsp;2 x10-4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2.44\u0026thinsp;\u0026plusmn;\u0026thinsp;1x 10\u0026thinsp;\u0026minus;\u0026thinsp;4\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eEthanol\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.16x10-9\u0026thinsp;\u0026plusmn;\u0026thinsp;1x10-12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.55 x10-4\u0026thinsp;\u0026plusmn;\u0026thinsp;3x10-6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.21x10-7\u0026thinsp;\u0026plusmn;\u0026thinsp;1x10-9\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec25\" class=\"Section2\"\u003e\u003ch2\u003e3.4. Tyrosinase Inhibition\u003c/h2\u003e\u003cp\u003eThe ethanol extract from \u003cem\u003eVatica diospyroides\u003c/em\u003e buds (CP extract) exhibited strong tyrosinase inhibition, achieving 90% inhibition at a concentration of 2.5 mg/mL, comparable to kojic acid (92.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5%) at the same concentration (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Buds were chosen for this assay based on their high phenolic and flavonoid content, as revealed by phytochemical screening, indicating their potential for significant bioactivity.\u003c/p\u003e\u003cp\u003e\u003cspan\u003e\u003cstrong\u003e3.5 Cytotoxicity\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003eThe antimicrobial activity of \u003cem\u003eVatica diospyroides\u003c/em\u003e ethanol bud extract (CP) was evaluated against Cutibacterium acnes at concentrations of 0.1 mg, 1 mg, and 10 mg, with clindamycin (0.002 mg) serving as a positive control (Fig. 4). At 0.1 mg, the CP extract produced a small inhibition zone, indicating limited activity. At 1 mg, the inhibition zone increased, demonstrating improved antimicrobial efficacy. The largest inhibition zone of 22.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.82 mm was observed at 10 mg, approaching the efficacy of clindamycin, which produced a larger inhibition zone due to its established potency. The results showed a dose-dependent increase in inhibition zones for the ethanol extract, with greater antimicrobial activity observed at higher concentrations. The CP extract at 10 mg exhibited notable inhibition, highlighting its potential activity against C. \u003cem\u003eacnes.\u003c/em\u003e\u003c/p\u003e\n\u003cdiv id=\"Sec26\" class=\"Section2\"\u003e\n \u003ch2\u003e3.6. Wound healing assay\u003c/h2\u003e\n \u003cp\u003eThe cytotoxicity of \u003cem\u003eVatica diospyroides\u003c/em\u003e ethanol bud extract (CP) was evaluated using the Sulforhodamine B (SRB) assay on human skin fibroblasts. The results indicated a concentration dependent effect on cell viability. At the highest concentration tested (1.0 mg/mL), CP-treated cells exhibited a viability rate of 75.3% \u0026plusmn; 3.2%, compared to 92.5% \u0026plusmn; 2.1% for cells treated with Vitamin C at the same concentration (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e). Cell viability remained above 70% across all tested concentrations of CP.\u003c/p\u003e\n \u003ch2\u003e3.7. Wound healing assay\u003c/h2\u003e\n \u003cp\u003eThe scratch assay demonstrated the wound-healing efficacy of \u003cem\u003eVatica diospyroides\u003c/em\u003e ethanol bud extract (CP) compared to control groups (DMEM and 10% DMSO) and Vitamin C as the positive control (Figs. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e and 7). The CP-treated group exhibited significant wound closure over time. At 6, 24, and 48 h, the CP-treated cells achieved wound closure rates of 47.01% \u0026plusmn; 0.61%, 58.25% \u0026plusmn;1.07%, and 46.70% \u0026plusmn; 0.64%, respectively. Vitamin C showed the highest wound closure rate at 87.51% \u0026plusmn; 2.27% after 48 h. Images captured at 0, 6, 24, and 48 h revealed consistent and significant migration of CP-treated cells towards the wound area, surpassing the control groups. By 48 h, the CP-treated group showed a nearly closed wound area comparable to the Vitamin C group.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eThe bioactivities of \u003cem\u003eVatica diospyroides\u003c/em\u003e flower extracts demonstrated multifunctional potential for cosmetic and therapeutic applications. Phytochemical screening confirmed the presence of key secondary metabolites, including phenolics, flavonoids, and terpenoids, particularly in the ethanol bud extracts, aligning with the observed bioactivities (Harborne, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Trease \u0026amp; Evans, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). The antioxidant capacity of the ethanol bud extract, with an IC50 of 1.16\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;9\u003c/sup\u003e\u0026plusmn;1\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;12\u003c/sup\u003e mg/mL, surpassed many plant-based antioxidants (Brand-Williams et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1995\u003c/span\u003e), while the 90% tyrosinase inhibition at 2.5 mg/mL matched kojic acid, highlighting its potential as a depigmenting agent (Briganti et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). Antimicrobial activity against Cutibacterium acnes was dose-dependent, with a maximum inhibition zone of 22.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.82 mm at 10 mg, indicating promise as a natural alternative for acne treatment (Srisawat et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Cytotoxicity assays showed moderate activity, maintaining over 70% fibroblast viability at all tested concentrations, supporting its safety for topical use (Vichai \u0026amp; Kirtikara, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). The wound healing assay revealed significant cell migration and wound closure rates, comparable to Vitamin C, demonstrating the extract's potential in regenerative skincare formulations (Trinh et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). These findings establish \u003cem\u003eV. diospyroides\u003c/em\u003e ethanol bud extract as a promising candidate for natural cosmetic applications, warranting further studies to optimize formulations and assess clinical efficacy.\u003c/p\u003e"},{"header":"5. Conclusions","content":"\u003cp\u003eThe study demonstrated that \u003cem\u003eVatica diospyroides\u003c/em\u003e ethanol bud extract is a valuable source of bioactive compounds with applications in natural cosmetics. Its robust antioxidant, tyrosinase inhibitory, antimicrobial, and wound-healing properties support its use in skincare formulations. While the extract showed moderate cytotoxicity, its safety profile within appropriate concentration ranges makes it suitable for topical application. Future studies should focus on isolating the active compounds, refining extraction techniques, and evaluating its clinical efficacy to fully realize its potential in sustainable cosmetic and therapeutic products.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eAcknowledgements\u003c/p\u003e\n\u003cp\u003eThe authors would like to thank the Cosmetic Science, Health \u0026amp; Anti-aging, Faculty of Science, Technology \u0026amp; Agriculture at Yala Rajabhat University and Mae Lan Learning Center.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;Fundings\u003c/p\u003e\n\u003cp\u003eThis research was supported by Yala Rajabhat University. [Grant number 4709851].\u003c/p\u003e\n\u003cp\u003eConflict of Interest\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no conflict of interest.\u003c/p\u003e\n\u003cp\u003eAuthor Contributions\u003c/p\u003e\n\u003cp\u003eN. Muhamad conceptualized the study, developed methodology, conducted investigations, performed formal analysis, wrote the original draft, administered the project, and acquired funding. L. Lateh contributed to conceptualization, data collection, and manuscript review. U. Tansom developed methodology, collected data, performed analysis, and reviewed the manuscript. P.S. Sinchai contributed to conceptualization, provided supervision, and reviewed the manuscript. T. Ninwijit provided supervision and reviewed the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003eEthics Approval\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThis study did not require ethics approval as it involved only plant material collection and in vitro assays. No human participants or animal subjects were involved.\u003c/p\u003e\n\u003cp\u003eConsent to Participate\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003eConsent to Publish\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003eAvailability of data and materials\u003c/p\u003e\n\u003cp\u003eThe datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAyoola, G. A., Coker, H. A. B., Adesegun, S. A., Adepoju-Bello, A. A., Obaweya, K., Ezennia, E. C., \u0026amp; Atangbayila, T. O. (2008). Phytochemical screening and antioxidant activities of some selected medicinal plants used for malaria therapy in Southwestern Nigeria. \u003cem\u003eTropical Journal of Pharmaceutical Research, 7\u003c/em\u003e(3), 1019–1024. https://doi.org/10.4314/tjpr.v7i3.14686\u003c/li\u003e\n \u003cli\u003eBauer, A. W., Kirby, W. M. M., Sherris, J. C., \u0026amp; Turck, M. (1966). Antibiotic susceptibility testing by a standardized single disk method. \u003cem\u003eAmerican Journal of Clinical Pathology, 45\u003c/em\u003e(4), 493–496. https://doi.org/10.1093/ajcp/45.4_ts.493\u003c/li\u003e\n \u003cli\u003eBrand-Williams, W., Cuvelier, M. E., \u0026amp; Berset, C. (1995). Use of a free radical method to evaluate antioxidant activity. \u003cem\u003eLWT \u003c/em\u003e\u003cem\u003e- \u003c/em\u003e\u003cem\u003eFood Science and Technology, 28\u003c/em\u003e(1), 25–30. https://doi.org/10.1016/S0023-6438(95)80008-5\u003c/li\u003e\n \u003cli\u003eBriganti, S., Camera, E., \u0026amp; Picardo, M. (2003). Chemical and instrumental approaches to treat hyperpigmentation. \u003cem\u003ePigment Cell Research, 16\u003c/em\u003e(2), 101–110. https://doi.org/10.1034/j.1600- 0749.2003.00006.x\u003c/li\u003e\n \u003cli\u003eCory, G., 2011. Scratch-Wound Assay. In: Wells, C., Parsons, M. (eds) Cell Migration. Methods in Molecular Biology. 769. Humana Press.\u003c/li\u003e\n \u003cli\u003eChothiphirat, A., Nittayaboon, K., Kanokwiroon, K., Srisawat, T., \u0026amp; Navakanitworakul, R. (2019). Anticancer potential of fruit extracts from \u003cem\u003eVatica diospyroides \u003c/em\u003eSymington Type SS and their effect on program cell death of cervical cancer cell lines. \u003cem\u003eTheScientificWorldJournal, 2019\u003c/em\u003e, 5491904. https://doi.org/10.1155/2019/5491904\u003c/li\u003e\n \u003cli\u003eChothiphirat, P., Narkkong, N. A., \u0026amp; Limpaiboon, T. (2019). Induction of apoptosis and autophagy in human cervical cancer cells by \u003cem\u003eVatica diospyroides \u003c/em\u003efruit extracts. \u003cem\u003eJournal of Medicinal Plants Research, 13\u003c/em\u003e(2), 23–30. https://doi.org/10.5897/JMPR2018.6684\u003c/li\u003e\n \u003cli\u003eGil-Martín, E., Forbes-Hernández, T., Romero, A., Cianciosi, D., Giampieri, F., \u0026amp; Battino, M. (2022). Influence of the extraction method on the recovery of bioactive phenolic compounds from food industry by-products. \u003cem\u003eFood Chemistry, 378\u003c/em\u003e, 131918. https://doi.org/10.1016/j.foodchem.2021.131918\u003c/li\u003e\n \u003cli\u003eGreen, L. C., Wagner, D. A., Glogowski, J., Skipper, P. L., Wishnok, J. S., \u0026amp; Tannenbaum, S. R. (1982). Analysis of nitrate, nitrite, and [15N] nitrate in biological fluids. \u003cem\u003eAnalytical Biochemistry,126\u003c/em\u003e(1), 131–138. https://doi.org/10.1016/0003-2697(82)90118-X\u003c/li\u003e\n \u003cli\u003eHarborne, J. B. (1998). \u003cem\u003ePhytochemical methods\u003c/em\u003e\u003cem\u003e: \u003c/em\u003e\u003cem\u003eA guide to modern techniques of plant analysis \u003c/em\u003e(3rd ed.). Springer. https://doi.org/10.1007/978-94-009-5921-7\u003c/li\u003e\n \u003cli\u003eHayouni, E. A., Abedrabba, M., Bouix, M., \u0026amp; Hamdi, M. (2007). The effects of solvents and extraction method on the phenolic contents and biological activities of Tunisian \u003cem\u003eQuercus coccifera L\u003c/em\u003e\u003cem\u003e. \u003c/em\u003eand \u003cem\u003eJuniperus phoenicea L\u003c/em\u003e\u003cem\u003e. \u003c/em\u003efruit extracts. \u003cem\u003eFood Chemistry, 105\u003c/em\u003e(3), 1126–1134. https://doi.org/10.1016/j.foodchem.2007.02.010\u003c/li\u003e\n \u003cli\u003eJutamanee, K., Chatuphonprasert, W., \u0026amp; Chaiyasut, C. (2016). Evaluation of anti-inflammatory and analgesic activities of ethanolic extracts from leaves and stems of \u003cem\u003eVatica diospyroides\u003c/em\u003e. \u003cem\u003eJournal of Medicinal Plants Research, 10\u003c/em\u003e(12), 232–238. https://doi.org/10.5897/JMPR2015.6043\u003c/li\u003e\n \u003cli\u003eKanokrat, N., Chaiyasut, C., \u0026amp; Sirithunyalug, J. (2022). Anti-inflammatory and antioxidant activities of Dipterocarpaceae plants: A comparative study. \u003cem\u003eAsian Journal of Pharmaceutical Sciences, 17\u003c/em\u003e(3), 225–231. https://doi.org/10.1016/j.ajps.2021.05.005\u003c/li\u003e\n \u003cli\u003eKhatib, S., Nerya, O., Musa, R., Shmuel, M., Tamir, S., \u0026amp; Vaya, J. (2005). Tyrosinase inhibition by tyrosol and its derivatives: Kinetics and structure-activity relationship. \u003cem\u003ePhytochemistry, 66\u003c/em\u003e(8),991–998. https://doi.org/10.1016/j.phytochem.2005.03.001\u003c/li\u003e\n \u003cli\u003eMuflihah, Y. M., Gollavelli, G., \u0026amp; Ling, Y. C. (2021). Correlation study of antioxidant activity with phenolic and flavonoid compounds in 12 Indonesian indigenous herbs. \u003cem\u003eAntioxidants, 10\u003c/em\u003e(10), 1530. https://doi.org/10.3390/antiox10101530\u003c/li\u003e\n \u003cli\u003eMusimun, C., Chuysongmuang, M., Permpoonpattana, P., Chumkaew, P., Sontikul, Y., Ummarat, N., \u0026amp; Srisawat, T. (2017). FACS analysis of bacterial responses to extracts of \u003cem\u003eVatica diospyroides \u003c/em\u003efruit show dose and time dependent induction patterns. \u003cem\u003eWalailak Journal of Science and Technology, 14\u003c/em\u003e(11), 883–891.\u003c/li\u003e\n \u003cli\u003eOhguchi, K., Tanaka, T., Ito, T., Iinuma, M., Matsumoto, K., Akao, Y., \u0026amp; Nozawa, Y. (2003). Inhibitory effects of resveratrol derivatives from Dipterocarpaceae plants on tyrosinase activity. \u003cem\u003eBioscience, Biotechnology, and Biochemistry, 67\u003c/em\u003e(7), 1587–1589. https://doi.org/10.1271/bbb.67.1587\u003c/li\u003e\n \u003cli\u003ePacker, L., Weber, S. U., \u0026amp; Rimbach, G. (2008). Molecular aspects of alpha-tocotrienol antioxidant action and cell signalling. \u003cem\u003eThe Journal of Nutritional Biochemistry, 12\u003c/em\u003e(1), 39–43. https://doi.org/10.1016/S0955-2863(00)00154-9\u003c/li\u003e\n \u003cli\u003ePandey, K. B., \u0026amp; Rizvi, S. I. (2009). Plant polyphenols as dietary antioxidants in human health and disease. \u003cem\u003eOxidative Medicine and Cellular Longevity, 2\u003c/em\u003e(5), 270–278. https://doi.org/10.4161/oxim.2.5.9498\u003c/li\u003e\n \u003cli\u003ePillaiyar, T., Manickam, M., \u0026amp; Namasivayam, V. (2017). Skin whitening agents: Medicinal chemistry perspective of tyrosinase inhibitors. \u003cem\u003eJournal of Enzyme Inhibition and Medicinal Chemistry, 32\u003c/em\u003e(1), 403–425. https://doi.org/10.1080/14756366.2016.1256882\u003c/li\u003e\n \u003cli\u003ePooma, R., \u0026amp; Newman, M. F. (2001). \u003cem\u003eDipterocarpaceae of Thailand\u003c/em\u003e\u003cem\u003e: \u003c/em\u003e\u003cem\u003eTaxonomy and conservation status \u003c/em\u003e(1st ed.). Royal Botanic Garden Edinburgh.\u003c/li\u003e\n \u003cli\u003eRai, R., McGibbon, A., \u0026amp; Chalabianloo, F. (2020). The role of plant-based antioxidants in delaying skin aging. \u003cem\u003eDermatologic Therapy, 33\u003c/em\u003e(6), e13491. https://doi.org/10.1111/dth.13491\u003c/li\u003e\n \u003cli\u003eRicciotti, E., \u0026amp; FitzGerald, G. A. (2011). Prostaglandins and inflammation. \u003cem\u003eArteriosclerosis,Thrombosis, and Vascular Biology, 31\u003c/em\u003e(5), 986–1000. https://doi.org/10.1161/ATVBAHA.110.207449\u003c/li\u003e\n \u003cli\u003eShahidi, F., \u0026amp; Zhong, Y. (2015). Measurement of antioxidant activity. \u003cem\u003eJournal of Functional Foods, 18\u003c/em\u003e(B), 757–781. https://doi.org/10.1016/j.jff.2015.01.047\u003c/li\u003e\n \u003cli\u003eSolano, F., Briganti, S., Picardo, M., \u0026amp; Ghanem, G. (2006). Hypopigmenting agents: An updated review on biological, chemical and clinical aspects. \u003cem\u003ePigment Cell Research, 19\u003c/em\u003e(6), 550–571. https://doi.org/10.1111/j.1600-0749.2006.00335.x\u003c/li\u003e\n \u003cli\u003eSrisawat, T., Phanthong, P., \u0026amp; Nualkaew, S. (2013). Cytotoxic and antimicrobial activities of \u003cem\u003eVatica diospyroides \u003c/em\u003eextracts. \u003cem\u003eAsian Pacific Journal of Tropical Biomedicine, 3\u003c/em\u003e(9), 688–692. https://doi.org/10.1016/S2221-1691(13)60141-0\u003c/li\u003e\n \u003cli\u003eSudsai, P., \u0026amp; Tewtrakul, S. (2013). Anti-inflammatory effects of phenolic compounds from medicinal plants. \u003cem\u003eJournal of Ethnopharmacology, 149\u003c/em\u003e(1), 217–220. https://doi.org/10.1016/j.jep.2013.06.014\u003c/li\u003e\n \u003cli\u003eTorres, E. A. S., Bazotte, R. B., Abdalla, D. S., Arcuri, J. C. F., \u0026amp; Rogero, M. M. (1987). Correlation between free radical scavenging activity and antioxidant activity in standard compounds. \u003cem\u003eInternational Journal of Pharmacology, 1\u003c/em\u003e(2), 86–94.\u003c/li\u003e\n \u003cli\u003eTrease, G. E., \u0026amp; Evans, W. C. (2002). \u003cem\u003ePharmacognosy \u003c/em\u003e(15th ed.). Saunders.\u003c/li\u003e\n \u003cli\u003eTrinh, X. T., Long, N. V., Van Anh, L. T., Nga, P. T., Giang, N. N., Chien, P. N., Nam, S. Y., \u0026amp; Heo, C. Y. (2022). A comprehensive review of natural compounds for wound healing: Targeting bioactivity perspective. \u003cem\u003eInternational Journal of Molecular Sciences, 23\u003c/em\u003e(17), 9573. https://doi.org/10.3390/ijms23179573\u003c/li\u003e\n \u003cli\u003eVichai, V., \u0026amp; Kirtikara, K. (2006). Sulforhodamine B colorimetric assay for cytotoxicity screening. \u003cem\u003eNature Protocols, 1\u003c/em\u003e(3), 1112–1116. https://doi.org/10.1038/nprot.2006.179\u003c/li\u003e\n \u003cli\u003eWojdyło, A., Oszmiański, J., \u0026amp; Czemerys, R. (2007). Antioxidant activity and phenolic compounds in 32 selected herbs. \u003cem\u003eFood Chemistry, 105\u003c/em\u003e(3), 940–949. https://doi.org/10.1016/j.foodchem.2007.04.038\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"applied-biochemistry-and-biotechnology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"abab","sideBox":"Learn more about [Applied Biochemistry and Biotechnology](https://www.springer.com/journal/12010)","snPcode":"12010","submissionUrl":"https://submission.nature.com/new-submission/12010/3","title":"Applied Biochemistry and Biotechnology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Vatica diospyroides, natural cosmetics, phytochemical screening, antioxidant activity, tyrosinase inhibition, wound healing","lastPublishedDoi":"10.21203/rs.3.rs-7158742/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7158742/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study investigates the phytochemical composition and bioactivities of \u003cem\u003eVatica diospyroides\u003c/em\u003e flower extracts, emphasizing their potential applications in natural product development. Phytochemical screening revealed the presence of alkaloids, phenolics, flavonoids, and terpenoids, with ethanol extracts showing the highest yield of bioactive compounds. The antioxidant activity of the ethanol bud extract was exceptional, with an IC50 value of 1.16\u0026times;10⁻⁹ mg/mL, surpassing many reported plant-based antioxidants. Anti-inflammatory effects were demonstrated by 70% nitric oxide inhibition in LPS-stimulated macrophages at 2.5 mg/mL, highlighting its potential for managing skin inflammation. Tyrosinase inhibition assays indicated 90% inhibition at 2.5 mg/mL, comparable to kojic acid, suggesting its suitability as a natural depigmenting agent. Antimicrobial activity against Propionibacterium acnes was dose-dependent, with significant inhibition at higher concentrations. In wound healing assays, the ethanol bud extract promoted fibroblast migration, achieving a 46.7% closure rate at 48 h, closely mirroring the effects of Vitamin C. These findings support the use of \u003cem\u003eV. diospyroides\u003c/em\u003e flower extracts as multifunctional natural ingredients in cosmetic formulations. Future studies should focus on optimizing extraction techniques, assessing stability in formulations, and validating efficacy through clinical trials to advance their integration into sustainable and eco-friendly skincare products.\u003c/p\u003e","manuscriptTitle":"Bioactivities and Phytochemical Potential of Vatica diospyroides Flower Extracts","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-30 11:27:06","doi":"10.21203/rs.3.rs-7158742/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"editorAssigned","content":"","date":"2025-08-28T08:19:51+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2025-08-07T10:17:13+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-07-28T06:46:11+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"Applied Biochemistry and Biotechnology","date":"2025-07-26T08:06:13+00:00","index":"","fulltext":""},{"type":"submitted","content":"Applied Biochemistry and Biotechnology","date":"2025-07-25T04:40:35+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"applied-biochemistry-and-biotechnology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"abab","sideBox":"Learn more about [Applied Biochemistry and Biotechnology](https://www.springer.com/journal/12010)","snPcode":"12010","submissionUrl":"https://submission.nature.com/new-submission/12010/3","title":"Applied Biochemistry and Biotechnology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"d22f8a80-3d98-46c2-8188-8a9916e28ca4","owner":[],"postedDate":"July 30th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-02-16T16:00:39+00:00","versionOfRecord":{"articleIdentity":"rs-7158742","link":"https://doi.org/10.1007/s12010-026-05603-2","journal":{"identity":"applied-biochemistry-and-biotechnology","isVorOnly":false,"title":"Applied Biochemistry and Biotechnology"},"publishedOn":"2026-02-13 15:57:04","publishedOnDateReadable":"February 13th, 2026"},"versionCreatedAt":"2025-07-30 11:27:06","video":"","vorDoi":"10.1007/s12010-026-05603-2","vorDoiUrl":"https://doi.org/10.1007/s12010-026-05603-2","workflowStages":[]},"version":"v1","identity":"rs-7158742","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7158742","identity":"rs-7158742","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}