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Its bioactive compounds possess antioxidant, antimicrobial, and anti-inflammatory properties, making it a promising candidate for multifunctional skincare formulations. Methods This study investigated the extraction of bioactive compounds from P. amboinicus leaves using microwave-assisted ethanol extraction. Two drying techniques—tray drying and freeze-drying—were compared to evaluate their impact on the extraction efficiency. The optimal extract (PF15), prepared using 15-minute microwave extraction and freeze-drying, was selected for further analysis. Bioactive content was assessed through quantification of caffeic acid, total phenolic content, and antioxidant activity via the DPPH assay. The antimicrobial activity of PF15 was tested against Staphylococcus aureus, Staphylococcus epidermidis, and Cutibacterium acnes. Anti-inflammatory potential was evaluated in LPS-stimulated human THP-1 macrophages by measuring cytokine production. Results The PF15 extract yielded the highest levels of bioactive compounds and demonstrated strong antioxidant activity. It exhibited significant antimicrobial effects against all tested skin pathogens. In the anti-inflammatory assay, PF15 significantly decreased pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) while upregulating the anti-inflammatory cytokine IL-10. The extract was formulated into a topical cream, which underwent accelerated stability testing over six heat-cool cycles. The cream remained stable with no signs of phase separation, discoloration, odor change, or microbial contamination, and maintained a pH of 5.5. Conclusions The PF15 extract of P. amboinicus demonstrates potent antioxidant, antimicrobial, and anti-inflammatory properties. Its successful incorporation into a stable cream formulation supports its potential as a multifunctional active ingredient in skincare products. These findings highlight P. amboinicus as a valuable natural source for the development of cosmetic formulations targeting oxidative stress, microbial infection, and inflammation. \" } { \"@context\": \"http://schema.org\", \"@type\": \"BreadcrumbList\", \"itemListElement\": [ { \"@type\": \"ListItem\", \"position\": \"1\", \"item\": { \"@id\": \"https://f1000research.com/\", \"name\": \"Home\" } }, { \"@type\": \"ListItem\", \"position\": \"2\", \"item\": { \"@id\": \"https://f1000research.com/browse/articles\", \"name\": \"Browse\" } }, { \"@type\": \"ListItem\", \"position\": \"3\", \"item\": { \"@id\": \"https://f1000research.com/articles/14-796/v1\", \"name\": \"Bioactive Effects of Plectranthus amboinicus Extract Using Microwave...\" } } ] } Home Browse Bioactive Effects of Plectranthus amboinicus Extract Using Microwave... ALL Metrics - Views Downloads Get PDF Get XML Cite How to cite this article Chinnahong C, U-Kong W, Rattanapot T et al. Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Techniques and Its Value Addition in Cosmeceutical Products [version 1; peer review: 1 approved, 1 not approved] . F1000Research 2025, 14 :796 ( https://doi.org/10.12688/f1000research.165030.1 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. Close Copy Citation Details Export Export Citation Sciwheel EndNote Ref. Manager Bibtex ProCite Sente EXPORT Select a format first Track Share ▬ ✚ Research Article Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Techniques and Its Value Addition in Cosmeceutical Products [version 1; peer review: 1 approved, 1 not approved] Chananan Chinnahong 1 , Warut U-Kong https://orcid.org/0000-0002-7972-6204 2 , Thiravat Rattanapot 2 , Chetsalit Hongnueng https://orcid.org/0009-0002-4651-2514 2 , Doungporn Amornlerdpison https://orcid.org/0000-0002-3429-1366 2,3 Chananan Chinnahong 1 , Warut U-Kong https://orcid.org/0000-0002-7972-6204 2 , [...] Thiravat Rattanapot 2 , Chetsalit Hongnueng https://orcid.org/0009-0002-4651-2514 2 , Doungporn Amornlerdpison https://orcid.org/0000-0002-3429-1366 2,3 PUBLISHED 18 Aug 2025 Author details Author details 1 Interdisciplinary Agriculture Program, Faculty of Agricultural Production,, Maejo University, Nong Han, Chiang Mai, Thailand 2 Center of Excellence in Agricultural Innovation for Graduate Entrepreneur, Maejo University, Nong Han, Chiang Mai, Thailand 3 Faculty of Fisheries Technology and Aquatic Resources, Maejo University, Nong Han, Chiang Mai, Thailand Chananan Chinnahong Roles: Data Curation, Formal Analysis, Investigation, Resources Warut U-Kong Roles: Investigation, Methodology, Validation, Writing – Original Draft Preparation Thiravat Rattanapot Roles: Methodology, Validation, Writing – Original Draft Preparation Chetsalit Hongnueng Roles: Formal Analysis, Methodology, Software, Writing – Original Draft Preparation Doungporn Amornlerdpison Roles: Conceptualization, Funding Acquisition, Project Administration, Resources, Supervision, Validation, Visualization, Writing – Review & Editing OPEN PEER REVIEW DETAILS REVIEWER STATUS This article is included in the Plant Science gateway. Abstract Background Plectranthus amboinicus is an aromatic herb known for its medicinal properties and is increasingly explored for cosmetic applications. Its bioactive compounds possess antioxidant, antimicrobial, and anti-inflammatory properties, making it a promising candidate for multifunctional skincare formulations. Methods This study investigated the extraction of bioactive compounds from P. amboinicus leaves using microwave-assisted ethanol extraction. Two drying techniques—tray drying and freeze-drying—were compared to evaluate their impact on the extraction efficiency. The optimal extract (PF15), prepared using 15-minute microwave extraction and freeze-drying, was selected for further analysis. Bioactive content was assessed through quantification of caffeic acid, total phenolic content, and antioxidant activity via the DPPH assay. The antimicrobial activity of PF15 was tested against Staphylococcus aureus , Staphylococcus epidermidis , and Cutibacterium acnes. Anti-inflammatory potential was evaluated in LPS-stimulated human THP-1 macrophages by measuring cytokine production. Results The PF15 extract yielded the highest levels of bioactive compounds and demonstrated strong antioxidant activity. It exhibited significant antimicrobial effects against all tested skin pathogens. In the anti-inflammatory assay, PF15 significantly decreased pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) while upregulating the anti-inflammatory cytokine IL-10. The extract was formulated into a topical cream, which underwent accelerated stability testing over six heat-cool cycles. The cream remained stable with no signs of phase separation, discoloration, odor change, or microbial contamination, and maintained a pH of 5.5. Conclusions The PF15 extract of P. amboinicus demonstrates potent antioxidant, antimicrobial, and anti-inflammatory properties. Its successful incorporation into a stable cream formulation supports its potential as a multifunctional active ingredient in skincare products. These findings highlight P. amboinicus as a valuable natural source for the development of cosmetic formulations targeting oxidative stress, microbial infection, and inflammation. READ ALL READ LESS Keywords Plectranthus amboinicus, microwave-assisted extraction, antioxidant activity, antimicrobial activity, anti-inflammatory activity Corresponding Author(s) Doungporn Amornlerdpison ( [email protected] ) Close Corresponding author: Doungporn Amornlerdpison Competing interests: No competing interests were disclosed. Grant information: The author(s) declared that no grants were involved in supporting this work. Copyright: © 2025 Chinnahong C et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. How to cite: Chinnahong C, U-Kong W, Rattanapot T et al. Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Techniques and Its Value Addition in Cosmeceutical Products [version 1; peer review: 1 approved, 1 not approved] . F1000Research 2025, 14 :796 ( https://doi.org/10.12688/f1000research.165030.1 ) First published: 18 Aug 2025, 14 :796 ( https://doi.org/10.12688/f1000research.165030.1 ) Latest published: 24 Jan 2026, 14 :796 ( https://doi.org/10.12688/f1000research.165030.3 )  There is a newer version of this article available. Suppress this message for one day. Introduction The global herbal cosmetic market has experienced consistent growth in recent years, driven by increasing consumer demand for natural and organic products. This trend reflects heightened awareness of the adverse effects associated with synthetic chemicals commonly used in cosmetic formulations, including allergic reactions, skin irritation, and potential long-term health risks. According to Custom Market Insights, 1 the global market value of herbal beauty products—which encompasses skincare, haircare, and fragrances for both men and women—was estimated at USD 78.9 billion in 2023 and is projected to reach USD 150.2 billion by 2032, with a compound annual growth rate (CAGR) of 6.5%. The application of plant extracts in cosmetic products has surged due to their diverse biological activities, such as antioxidant, anti-inflammatory, UV-protective, anti-aging, and anti-acne effects. 2 These benefits, coupled with growing concerns over the safety of synthetic ingredients, have prompted a shift in consumer preference towards natural alternatives. Medicinal plants offer a promising and safer source of bioactive compounds for use in cosmetic formulations. In the context of Thailand, the integration of native medicinal plants into cosmetic development presents an opportunity to enhance the value and global competitiveness of local herbal resources. Various plant parts—including leaves, flowers, fruits, stems, and roots—are known to exhibit a wide range of bioactivities, including antibacterial, antifungal, and yeast-inhibitory effects, along with properties that support skin nourishment and restoration. 3 Plectranthus amboinicus , a perennial herb indigenous to Southeast Asia, has been widely utilized in traditional culinary practices and herbal medicine. Its leaves are rich in numerous bioactive constituents, such as essential oils (e.g., carvacrol, thymol, γ-terpinene) and phenolic compounds (e.g., caffeic acid, quercetin, ursolic acid, and rosmarinic acid), which exhibit potent antioxidant, antimicrobial, and anti-inflammatory properties. Additionally, these compounds have demonstrated other pharmacological activities, including anticancer potential. 4 , 5 A recent study by Ref. 6 further highlighted the antimicrobial efficacy of P. amboinicus leaf extract against pathogens such as Staphylococcus aureus , Escherichia coli , Pseudomonas aeruginosa , Candida albicans , and the acne-associated bacterium Cutibacterium acnes , reinforcing its potential application in cosmetic products. Various techniques have been employed to extract bioactive compounds from plants, including maceration, Soxhlet extraction, supercritical CO 2 extraction, hydrothermal processing, and ultrasonic-assisted extraction. Despite their widespread use, these conventional methods often involve extended processing times, low extraction efficiency, and high operational costs. 7 Recently, microwave-assisted extraction (MAE) using 50% ethanol has emerged as a promising alternative. This technique offers several advantages, including shorter extraction time, higher yield and concentration of bioactive compounds, and reduced solvent consumption. 8 Moreover, MAE maintains the integrity and efficacy of extracts while ensuring safety and environmental sustainability, making it well-suited for industrial and commercial-scale applications. 9 , 10 Accordingly, the present study aimed to optimize the extraction of bioactive compounds from P. amboinicus leaves using microwave-assisted ethanol extraction. This was followed by a comparative evaluation of two drying techniques and the assessment of the extract’s antioxidant, antimicrobial, and anti-inflammatory activities. The final objective was to incorporate the extract into a prototype cosmetic formulation, such as an anti-acne or anti-inflammatory cream. The outcomes of this research are expected to enhance the commercial value of Thai medicinal herbs and promote the sustainable cultivation of P. amboinicus , thereby generating economic benefits for local communities. Materials and methods Extraction process optimization for Plectranthus amboinicus leaves Sample collection and preparation Plectranthus amboinicus leaves were collected from the Maejo area, San Sai District, Chiang Mai Province, Thailand. The leaves were washed with distilled water and dried in a hot air oven at 60 °C for 24 hours. Once dried, they were ground into a fine powder. Three separate 20-gram portions of the powdered leaves were weighed and placed into round-bottom flasks for extraction. Microwave-assisted extraction procedure Bioactive compounds were extracted using microwave-assisted extraction (MAE) at a power of 450 watts, using 200 mL of 50% ethanol as the solvent. The extraction was conducted at three different time intervals: 10, 15, and 20 minutes. 11 The extracts were filtered using Whatman No.1 filter paper, and the solvents were partially evaporated under reduced pressure using a rotary evaporator. Extracts were then stored at −40 °C for further analysis. Drying methods and yield determination The filtered extracts were subjected to two drying techniques: freeze-drying (PF) and tray-drying (PT). The percentage yield (% yield) was calculated using the formula: % Yield = ( Weight of Dried Extract / Weight of Starting Material ) × 100 Chemical characterization Caffeic acid quantification by HPLC Extracts from MAE (PT10, PT15, PT20, PF10, PF15, PF20) were analyzed for caffeic acid content. Each 0.05 g sample was dissolved in ethanol and diluted to 50 mL. Solutions were filtered through a 0.45 μm membrane filter and placed into 2 mL vials. High-performance liquid chromatography (HPLC) was used for quantification. Conditions are listed in Table 1 . Table 1. HPLC conditions (FLEXAR™ LC System, PerkinElmer). Condition Column Brownlee Analytical C18, 150 × 4.60 mm, 5.0 μm Mobile Phase (A) 0.1% Phosphoric acid in water, (B) Methanol (80:20) Flow Rate 0.3 mL/min Injection Temperature 40 °C Detector Photodiode array detector (PDA), 275 nm Injection Volume 5 μL Mode Isocratic elution Total Phenolic Content (TPC) determination TPC was measured using the Folin–Ciocalteu method on PF10, PF15, and PF20 samples. Extracts (200 μL) were mixed with 1,000 μL of Folin–Ciocalteu reagent and 800 μL of 7.5% sodium carbonate. After incubation at room temperature for 60 minutes, absorbance was measured at 765 nm. A gallic acid standard curve (16–250 mg/L) was used to calculate phenolic content, reported as mg gallic acid equivalents per gram (mg GAE/g extract). 12 , 13 Biological activity assessment Antioxidant activity (DPPH assay) The antioxidant activity of PF10, PF15, and PF20 was assessed using the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging method. 14 Various concentrations of extract (0.125–4 mg/mL) and L-ascorbic acid (12–400 μg/mL) were tested. After mixing 50 μL of each sample with 100 μL DPPH (2,000 μM), the reaction was incubated in darkness for 30 minutes. Absorbance was read at 517 nm. Scavenging activity (%) was calculated as: % DPPH = [ ( A _ control − A _ sample ) / A _ control ] × 100 The IC 50 value, indicating the concentration required to scavenge 50% of DPPH radicals, was used to compare antioxidant potential. Antimicrobial activity Bacterial preparation PF15 was tested against Staphylococcus aureus (ATCC 25932), Staphylococcus epidermidis (ATCC 14990), and Cutibacterium acnes (ATCC 6919). Bacteria were cultured in nutrient broth at 37 °C for 12–18 hours and adjusted to a 0.5 McFarland standard, giving an approximate cell density of 1.5 × 10 8 CFU/mL. Extract preparation for antimicrobial testing A 1,000 mg/mL stock solution of PF15 was prepared in sterile distilled water, filtered through a 0.2 μm membrane and used for antimicrobial testing. Disc diffusion assay Sterile discs were loaded with 10 μL of extract, dried, and placed on agar plates seeded with test bacteria. Sterile water and ampicillin (10 μg/disc) served as negative and positive controls. Plates were incubated at 37 °C for 24 hours and inhibition zones were measured. 15 MIC determination (Broth microdilution) PF15 was serially diluted in 96-well plates (1,000–0.48 mg/mL). Wells were inoculated with bacteria, and plates incubated at 37 °C for 24 hours. The MIC (Minimal Inhibitory Concentration ) was the lowest concentration with no visible bacterial growth. 15 MBC determination (Drop plate method) Aliquots from each MIC well were plated on nutrient agar. After incubation, the MBC (Minimal Bactericidal Concentration was the lowest concentration at which no bacterial colonies formed. 15 Anti-inflammatory activity assay THP-1 Cell culture and differentiation THP-1 monocytes (ATCC TIB-202) were cultured in RPMI-1640 medium with 10% FBS and 1% antibiotics. Differentiation into macrophages was induced using 50 ng/mL PMA for 48 hours, followed by 24 hours in fresh medium. 16 Cell viability assay Differentiated cells were treated with PF15 extract (0–200 μg/mL) and 0.5 μg/mL LPS (lipopolysaccharides) for 24 hours. PrestoBlue ® reagent was added, and absorbance was measured at 570 nm to assess cell viability. 17 Cytokine measurement by ELISA Cells were treated with PF15 (12, 25, and 50 μg/mL) along with LPS. Supernatants were collected after 24 hours and analyzed for TNF-α, IL-1β, IL-6, and IL-10 using ELISA kits (Thermo Fisher). Results represent means ± standard deviation (SD) from triplicate experiments. Product development and evaluation Cream Formulation with PF15 An anti-acne cream was formulated using the MIC concentration of PF15. The aqueous phase containing PF15 was heated to 70 °C, combined with the oil phase, and emulsified for 30 minutes. The cream was cooled to 35–40 °C, mixed until homogeneous, and stored for further analysis. Stability and safety evaluation Accelerated stability testing The cream underwent 6 thermal cycles (4 °C for 24 h and 45 °C for 24 h) to assess changes in pH, odor, color, texture, and phase separation. 18 Antimicrobial activity of the final product Using the disc diffusion method, antimicrobial activity was evaluated against S. aureus , S. epidermidis , and C. acnes. Cream without extract served as a negative control. Microbial contamination testing Aerobic Plate Count (APC) Cream samples were diluted and plated on plate count agar. Colonies were counted after 24 hours at 37 °C, reported as CFU/g. Yeast and mold contamination Samples were plated on Dichloran Rose Bengal Chloramphenicol agar after serial dilution. Colonies were counted after 24 hours to determine CFU/g. Escherichia coli detection Diluted samples were cultured in Lauryl Tryptose broth and incubated. Gas formation indicated presumptive E. coli. Confirmation was done using EC Broth and MPN methodology. Staphylococcus aureus detection Samples were diluted and plated on Mannitol Salt agar. Colony morphology and color change were used for identification. CFU/g was reported. 19 Salmonella spp. detection Samples were pre-enriched in Tryptone Soya broth, streaked onto SS Agar, and incubated for 24 hours. Growth consistent with Salmonella spp. was recorded as “present” or “absent” in 25 g of sample. 20 Statistical analysis All results are presented as mean ± SD. Statistical significance was assessed using one-way ANOVA, followed by Duncan’s Multiple Range Test. A p -value < 0.05 was considered significant. Data analysis was conducted using GraphPad Prism version 9.0.0.121. Results and Discussion Optimal extraction and physical characteristics of Plectranthus amboinicus extracts Microwave-assisted extraction (MAE) using 50% ethanol for 10, 15, and 20 minutes, followed by either tray drying (PT10, PT15, PT20) or freeze-drying (PF10, PF15, PF20), yielded extracts with distinct physical characteristics and varying efficiencies. Tray-dried extracts produced yields of 22.4, 23.5, and 22.5 g, respectively, demonstrating consistent extraction efficiency across the time intervals. These values are in alignment with the findings of, 21 which reported diminishing returns with extended extraction beyond optimal durations. The resulting tray-dried powders were dark green in color, indicating good retention of chlorophyll, flavonoids, and phenolic compounds ( Figure 1 ). Conversely, freeze-dried extracts yielded slightly lower quantities—19.8, 21.8, and 21.0 g for PF10, PF15, and PF20, respectively—yet retained a fine, homogeneous powder with vibrant green coloration. Despite lower yields, freeze-drying was effective in preserving thermolabile bioactives and improving powder texture. These visual differences in texture and color between drying methods ( Figure 2 ) highlight the critical influence of post-extraction drying on the final product characteristics, with freeze-drying offering superior preservation at the expense of yield. 22 Figure 1. Physical characteristics of P. amboinicus extracts obtained by tray drying and freeze-drying. Figure 2. Comparative yield of P. amboinicus extracts by drying method and extraction time. Caffeic acid content analysis Tray-Dried Extracts (PT) High-performance liquid chromatography (HPLC) analysis confirmed the presence of caffeic acid in all tray-dried extracts. The retention times observed for PT10, PT15, and PT20 were 13.94, 12.19, and 13.92 minutes, respectively, with corresponding concentrations of 1.04, 1.07, and 1.03 mg/g ( Figure 3b-d ). These results reflect consistent caffeic acid retention across varying extraction times, with PT15 slightly outperforming others. However, the overall concentration remained modest, suggesting that tray drying may limit phenolic preservation. Figure 3. HPLC chromatograms of caffeic acid standard and tray-dried samples. Freeze-dried extracts (PF) Caffeic acid content in freeze-dried extracts was significantly higher: 3.15, 3.33, and 3.29 mg/g for PF10, PF15, and PF20, respectively ( Figure 4b-d ). Retention times were consistent with the standard, confirming accurate identification. PF15 exhibited the highest caffeic acid content, indicating that 15 minutes of MAE under freeze-drying conditions is optimal for phenolic preservation. The increased levels of caffeic acid in PF samples may be attributed to the lower thermal stress during drying, which minimizes degradation of sensitive compounds. 21 Figure 4. HPLC chromatograms of caffeic acid standard and freeze-dried samples. The comparative analysis ( Figure 5 ) clearly illustrates that freeze-drying consistently outperformed tray drying in preserving caffeic acid content across all extraction durations. This emphasizes the role of gentle drying techniques in maintaining the chemical integrity of phytochemicals, supporting the use of freeze-drying for applications targeting high bioactive potency. Figure 5. Quantitative comparison of caffeic acid content in PT vs. PF samples at different extraction times. Total Phenolic Content (TPC) evaluation The TPC was highest in PF15, recorded at 70.46 ± 0.49 mg GAE/g extract, followed by PF10 and PF20. The differences were statistically significant ( p < 0.05), indicating that extraction time had a measurable impact on phenolic recovery ( Figure 6 ). Notably, the 15-minute extraction (PF15) demonstrated optimal balance between yield and stability, aligning with earlier research indicating that mid-range MAE durations improve phenolic extraction without inducing degradation. 23 , 21 These results collectively underscore the effectiveness of microwave-assisted extraction at 15 minutes in conjunction with freeze-drying as the most favorable combination for preserving caffeic acid and phenolic content in P. amboinicus leaf extracts. The application of this optimized method is particularly advantageous in the development of high-performance cosmeceutical products where antioxidant activity is desired. Figure 6. Total phenolic content in PF10, PF15, and PF20 samples. Biological activities of Plectranthus amboinicus leaf extracts (PF15) Antioxidant activity The Plectranthus amboinicus leaf extract obtained under optimized conditions (PF15) demonstrated significant antioxidant activity, as assessed using the DPPH radical scavenging assay. The extract exhibited a markedly low IC 50 value, indicating a strong ability to neutralize free radicals. Among the three extraction durations tested—10, 15, and 20 minutes—the PF15 extract showed the most potent antioxidant effect, with a statistically significant difference in IC 50 values compared to PF10 and PF20 ( p < 0.05) ( Figure 7 ). These findings align with the work of, 24 who reported the antioxidant potential of P. amboinicus. The enhanced activity at 15 minutes suggests that this extraction time enables maximal release and preservation of antioxidant phenolic compounds, including caffeic acid and other flavonoids. This result underscores the importance of extraction time optimization in maximizing the functional properties of herbal extracts. Such antioxidant efficacy supports the potential use of P. amboinicus extract in cosmetic formulations targeting oxidative stress-related skin issues, such as aging and environmental damage. Figure 7. IC 50 values for DPPH radical scavenging activity of P. amboinicus leaf extracts (PF10, PF15, PF20). Antimicrobial activity of PF15 Inhibition against skin pathogens The antimicrobial potential of PF15 was evaluated using the paper disc diffusion method. At a concentration of 1,000 mg/mL, PF15 exhibited clear zones of inhibition against S. aureus , S. epidermidis , and C. acnes. The diameter of the inhibition zones was significantly larger than those of the negative control (distilled water), indicating strong antibacterial efficacy ( Table 2 ). These results support the growing evidence that P. amboinicus possesses natural antimicrobial compounds effective against common skin pathogens. Its ability to inhibit C. acnes , a key contributor to acne development, reinforces its potential application in dermatological formulations such as anti-acne creams and antimicrobial cosmetics. 23 Table 2. Antibacterial activity of P. amboinicus leaf extract (PF15) against selected skin pathogens. Sample inhibition zone diameter (mm) S. aureus S. eipidermidis C. acnes PF5 13.33 ± 0.58 13.33 ± 0.58 15.33 ± 0.58 Ampicillin 27.67 ± 0.58 30.00 ± 1.00 27.67 ± 0.58 Distilled water - - - Minimum Inhibitory and Bactericidal Concentrations (MIC & MBC) The MIC and MBC values of the PF15 extract were determined using a broth microdilution assay. Results indicated that the MIC values against S. aureus , S. epidermidis , and C. acnes were 7.81, 3.91, and 3.91 mg/mL, respectively, while the corresponding MBC values were 31.25, 15.63, and 15.63 mg/mL ( Table 3 ). These data demonstrate that PF15 not only inhibits bacterial growth but also exhibits bactericidal effects at relatively low concentrations, particularly against C. acnes , a clinically relevant acne pathogen. The observed antibacterial potency is consistent with prior reports on the antimicrobial activities of P. amboinicus. 21 , 25 , 26 These findings provide strong support for the use of this plant extract in the formulation of herbal-based skin care products designed to prevent or treat bacterial skin infections. Table 3. MIC and MBC of P. amboinicus leaf extract (PF15) against selected bacteria. Sample MIC (mg/ml) MBC (mg/ml) S. aureus S. eipidermidis C. acne S. aureus S. eipidermidis C. acne PF15 7.81 3.91 3.91 31.25 15.63 15.63 Ampicillin 0.01 0.01 0.01 0.01 0.01 0.01 Distilled water - - - - - - Anti-inflammatory activity of PF15 Effect on THP-1 cell viability The cytotoxicity of PF15 was assessed in human THP-1 monocytes using the PrestoBlue ® viability assay. As illustrated in Figure 8 , PF15 at concentrations ranging from 6 to 50 μg/mL did not significantly reduce cell viability compared to the untreated control group. Cell viability remained above 80% at all tested concentrations, indicating that PF15 was non-cytotoxic under the experimental conditions. A statistically significant increase in viability was observed at certain concentrations ( p < 0.01 and p < 0.001), suggesting a potential dose-dependent proliferative or protective effect on THP-1 cells. Figure 8. Effect of P. amboinicus leaf extract (PF15) on THP-1 cell viability measured using the PrestoBlue ® assay. Data represents the mean ± SD from four independent experiments. * p < 0.01 and ** p < 0.001 compared to control (C) group. Modulation of cytokine secretion in LPS-stimulated THP-1 macrophages To evaluate the anti-inflammatory potential of PF15, the levels of pro- and anti-inflammatory cytokines were quantified in lipopolysaccharide (LPS)-stimulated THP-1 macrophages following treatment with PF15 at concentrations of 12, 25, and 50 μg/mL. LPS stimulation significantly increased the secretion of TNF-α, IL-1β, and IL-6 compared to the untreated control ( ## p < 0.001). However, treatment with PF15 significantly reduced the levels of these pro-inflammatory cytokines in a dose-dependent manner (* p < 0.05 and ** p < 0.001), as shown in Figure 9 . In contrast, the level of IL-10, a key anti-inflammatory cytokine, was significantly upregulated upon PF15 treatment, suggesting an immunomodulatory shift favoring resolution of inflammation. These results demonstrate that PF15 effectively suppresses pro-inflammatory signaling while enhancing anti-inflammatory responses in macrophages. Figure 9. Effects of PF15 on cytokine secretion in LPS-stimulated THP-1 macrophages. ELISA was used to quantify TNF-α (a), IL-1β (b), IL-6 (c), and IL-10 (d). Data are expressed as mean ± SD (n = 3). ## p < 0.001 vs. control (C); * p < 0.05, ** p < 0.001 vs. LPS group. The IL-6/IL-10 ratio serves as a reliable biomarker for evaluating the balance between pro- and anti-inflammatory responses. As depicted in Figure 10 , LPS stimulation significantly elevated the IL-6/IL-10 ratio, indicating a dominant pro-inflammatory state. Treatment with PF15 significantly reduced this ratio in a dose-dependent manner, suggesting a shift toward an anti-inflammatory phenotype and restoration of immune homeostasis. Figure 10. Effect of PF15 on IL-6/IL-10 ratio in LPS-stimulated THP-1 macrophages. Values represent mean ± SD from three independent experiments. ## p < 0.001 vs. control; * p < 0.01 and ** p < 0.001 vs. LPS group. The results of this study provide robust evidence that P. amboinicus leaf extract (PF15) exhibits significant anti-inflammatory activity through multiple mechanisms. The extract not only suppressed key pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) but also enhanced the production of IL-10, a cytokine critical for immune regulation and inflammation resolution. These findings are consistent with previous studies on plant-derived bioactive compounds exhibiting immunomodulatory effects in macrophage models. 27 , 28 The observed inhibition of TNF-α and IL-6 aligns with established mechanisms of anti-inflammatory plant compounds that interfere with Toll-like receptor (TLR)-mediated signaling pathways. 29 Suppression of IL-6 is particularly noteworthy, as its overexpression is implicated in the pathogenesis of chronic inflammatory diseases. 30 The upregulation of IL-10 further supports the role of PF15 in promoting an anti-inflammatory environment, a mechanism also observed in other herbal interventions. 31 , 32 Importantly, the decrease in the IL-6/IL-10 ratio suggests a rebalancing of immune responses, indicative of improved inflammatory resolution. 33 Similar regulatory effects have been reported in studies involving flavonoids and polyphenols, 16 , 17 highlighting the potential of PF15 as a natural anti-inflammatory agent. Formulation and development of a topical product containing PF15 Prototype cream formulation and evaluation A topical cream was formulated using PF15 and compared against a control formulation lacking the extract ( Figure 11 ). The extract-based cream displayed a light green color, smooth consistency, and favorable spreadability. It emitted a mild, herbal-minty scent, characteristic of the extract, and exhibited no signs of phase separation, sedimentation, or instability ( Table 4 ). The measured pH of 5.5 aligns with the optimal range for topical applications and skin compatibility. 34 These physical and organoleptic properties indicate the successful incorporation of P. amboinicus extract into a stable cosmetic matrix with potential applications in acne treatment and skin care product development. 35 Table 4. Physical properties of topical cream formulations. Product Texture Color Phase separation Fragrance pH Based cream Well-dispersed White None Cream-specific 5.5 PF15 cream Well-dispersed Light green None Minty/herbal 5.5 Figure 11. Cream formulation containing PF15 compared to based cream. Accelerated stability assessment of PF15 cream The PF15 cream underwent six thermal cycles (alternating storage at 4 ± 1 °C and 45 ± 1 °C) to simulate long-term environmental stress. The cream maintained a consistent texture, color, and fragrance across all cycles, with no evidence of phase separation or degradation. The pH remained stable at 5.5 throughout the testing period, confirming the formulation’s stability under extreme storage conditions. 36 – 38 Antibacterial efficacy of formulated PF15 cream Following accelerated stability testing, the PF15 cream retained its antibacterial activity against S. aureus , S. epidermidis , and C. acnes. The inhibition zones measured 11± 1.00 mm, 12± 0.82 mm, and 12± 0.82 mm (Mean ± SD, n = 3) respectively ( Figure 12 ), confirming that the antimicrobial properties of the extract were not compromised during storage. These results support its efficacy as a topical agent targeting acne-associated pathogens. 39 , 40 Figure 12. Antibacterial activity of PF15 cream post-stability testing, assessed using the paper disc diffusion method. Microbial contamination assessment Microbiological testing of the PF15 cream was conducted in compliance with the Thai Cosmetics standard and 41 The cream was assessed for aerobic plate count (APC), yeast and mold contamination, and the presence of Escherichia coli , Staphylococcus aureus , and Salmonella spp. All results were within the acceptable limits, confirming the microbiological safety of the final product ( Table 5 ). Table 5. Microbial contamination test results of PF15 cream. Microbial group Test result Unit Standard limit Aerobic plate count Not detected CFU/g <10 3 CFU/g Yeasts and molds Not detected CFU/g <10 3 CFU/g E. coli <3 MPN/g <10 MPN/g S. aureus Not detected CFU/g Not detected Salmonella spp. Not detected in 25 g Not detected These findings confirm that the PF15 cream meets safety standards for cosmetic use, is free from pathogenic microorganisms, and is suitable for topical applications. Safety evaluations based on FDA guidelines—including cytotoxicity and microbial contamination assessments—indicate that the formulation is safe for consumer use. This aligns with previous safety studies of P. amboinicus in cosmetic applications. 42 The successful formulation of a stable, effective, and safe cream containing P. amboinicus leaf extract (PF15) demonstrates the practical potential of this plant as a bioactive ingredient in dermatological and cosmeceutical products. The extract retained its biological properties post-formulation and post-stability testing, making it a promising candidate for anti-acne, antimicrobial, and anti-inflammatory skincare applications. Conclusion This study provides compelling evidence that P. amboinicus leaf extract, obtained through microwave-assisted extraction using 50% ethanol and stabilized via freeze-drying, is a rich source of bioactive compounds—particularly caffeic acid and total phenolics. The extract exhibited multifaceted biological activities, including potent antioxidant capacity, broad-spectrum antimicrobial effects against S. aureus , S. epidermidis , and C. acnes , and significant anti-inflammatory properties through the downregulation of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6). These findings support the potential use of P. amboinicus extract as a natural anti-inflammatory and antimicrobial agent suitable for incorporation into cosmetic and dermatological formulations. Importantly, the extract demonstrated excellent physicochemical stability and microbiological safety when formulated into a topical cream, retaining both efficacy and formulation integrity after accelerated stability testing. Further investigations—including in vivo efficacy studies and mechanistic analyses at the molecular level—are recommended to confirm the therapeutic relevance and elucidate the pathways involved. Overall, this research highlights the strong potential of P. amboinicus as a multifunctional, safe, and stable active ingredient in the development of innovative skincare products, particularly those targeting inflammation, acne, and oxidative stress-related skin conditions. Institutional review board Ethical approval and consent were not required. AI use disclosure In accordance with the Taylor & Francis AI Policy, generative AI tools (ChatGPT 4.0, OpenAI) were used solely for language editing and phrasing suggestions. All usage was conducted under full human supervision. Data availability The data that support the findings of this study are openly available from Zenodo at: https://zenodo.org/record/15796465 (doi: 10.5281/zenodo.15796465 ). This dataset was made available under the Creative Commons Attribution 4.0 International (CC BY 4.0) license. 43 Acknowledgments The authors express their sincere gratitude to the Center of Excellence in Agricultural Innovation for Graduate Entrepreneurs, Maejo University, for providing laboratory facilities, equipment, and technical support. Their assistance greatly contributed to the successful completion of this study. References 1. Custom Market Insights: Herbal beauty and personal care products market size, share, and trends analysis report by product (skin care, hair care, fragrance), by gender, by region, and segment forecasts, 2023–2032.2024 [cited 2025 Apr 12]. 2. Tang Y, Wang X: Applications of plant-based materials in cosmetics IntechOpen; 2025. 3. 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Publisher Full Text Comments on this article Comments (0) Version 3 VERSION 3 PUBLISHED 18 Aug 2025 ADD YOUR COMMENT Comment Author details Author details 1 Interdisciplinary Agriculture Program, Faculty of Agricultural Production,, Maejo University, Nong Han, Chiang Mai, Thailand 2 Center of Excellence in Agricultural Innovation for Graduate Entrepreneur, Maejo University, Nong Han, Chiang Mai, Thailand 3 Faculty of Fisheries Technology and Aquatic Resources, Maejo University, Nong Han, Chiang Mai, Thailand Chananan Chinnahong Roles: Data Curation, Formal Analysis, Investigation, Resources Warut U-Kong Roles: Investigation, Methodology, Validation, Writing – Original Draft Preparation Thiravat Rattanapot Roles: Methodology, Validation, Writing – Original Draft Preparation Chetsalit Hongnueng Roles: Formal Analysis, Methodology, Software, Writing – Original Draft Preparation Doungporn Amornlerdpison Roles: Conceptualization, Funding Acquisition, Project Administration, Resources, Supervision, Validation, Visualization, Writing – Review & Editing Competing interests No competing interests were disclosed. Grant information The author(s) declared that no grants were involved in supporting this work. Article Versions (3) version 3 Revised Published: 24 Jan 2026, 14:796 https://doi.org/10.12688/f1000research.165030.3 version 2 Revised Published: 13 Oct 2025, 14:796 https://doi.org/10.12688/f1000research.165030.2 version 1 Published: 18 Aug 2025, 14:796 https://doi.org/10.12688/f1000research.165030.1 Copyright © 2025 Chinnahong C et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Download Export To Sciwheel Bibtex EndNote ProCite Ref. Manager (RIS) Sente metrics Views Downloads F1000Research - - PubMed Central info_outline Data from PMC are received and updated monthly. - - Citations open_in_new 0 open_in_new 0 open_in_new SEE MORE DETAILS CITE how to cite this article Chinnahong C, U-Kong W, Rattanapot T et al. Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Techniques and Its Value Addition in Cosmeceutical Products [version 1; peer review: 1 approved, 1 not approved] . F1000Research 2025, 14 :796 ( https://doi.org/10.12688/f1000research.165030.1 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS track receive updates on this article Track an article to receive email alerts on any updates to this article. TRACK THIS ARTICLE Share Open Peer Review Current Reviewer Status: ? Key to Reviewer Statuses VIEW HIDE Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Version 1 VERSION 1 PUBLISHED 18 Aug 2025 Views 0 Cite How to cite this report: Khat-Udomkiri N. Reviewer Report For: Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Techniques and Its Value Addition in Cosmeceutical Products [version 1; peer review: 1 approved, 1 not approved] . F1000Research 2025, 14 :796 ( https://doi.org/10.5256/f1000research.181624.r413222 ) The direct URL for this report is: https://f1000research.com/articles/14-796/v1#referee-response-413222 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 27 Sep 2025 Nuntawat Khat-Udomkiri , Mae Fah Luang University, Mueang Chiang Rai, Chiang Rai, Thailand Not Approved VIEWS 0 https://doi.org/10.5256/f1000research.181624.r413222 Thank you for the opportunity to review this manuscript. The study of microwave-assisted extraction (MAE) from Plectranthus amboinicus for cosmeceutical use is topical and fits the journal scope, but the manuscript in its current form lacks novelty, methodological detail, and ... Continue reading READ ALL Thank you for the opportunity to review this manuscript. The study of microwave-assisted extraction (MAE) from Plectranthus amboinicus for cosmeceutical use is topical and fits the journal scope, but the manuscript in its current form lacks novelty, methodological detail, and sufficient experimental rigor. Below I list the main weaknesses and give concrete, actionable recommendations to improve clarity, reproducibility and scientific impact. 1. You must cite and discuss prior MAE work on P. amboinicus (e.g., the paper you identified: DOI: 10.3303/CET1972067). Clearly state how your study differs (new endpoints, different solvent system, biological assays, formulation work, scale-up considerations, etc.). Right now the manuscript reads like a repeat of existing work. 2. Do not claim you “optimized” extraction conditions unless a formal design of experiments (e.g., RSM, factorial) or an explicit optimization protocol was applied. If you did not run an optimization design, rephrase to “screened”, “evaluated” or “investigated”. 3. Temperature is critical for MAE outcomes and must be reported for every condition. Report temperature profiles for all MAE runs. 4. Explain the basis for selecting caffeic acid as the principal marker. Was it the most abundant compound in the extract, the most bioactive, or selected for analytical convenience? 5. Natural extracts are mixtures — identify and quantify multiple major phenolics, not only one. Provide HPLC chromatograms and quantification for at least the top 3–5 phenolic compounds. 6. Provide a full cream formulation (all ingredients, % w/w), the exact amount of extract incorporated, and the rationale for the selected concentration(s). 7. You state extract was included during emulsification: explain and justify this choice. Phenolic compounds is heat-sensitive; if used in a heated phase, show data proving it remains active after processing. 8. You cannot assert “no degradation” based on visual inspection alone. Provide objective stability data: HPLC profiles (initial vs after processing and after stability cycles), TPC, and at least one chemical marker quantified before/after heating and after storage cycles. Replace subjective color assessments with ΔE measurements and report ΔE for color change. 9. DPPH must include a positive control (e.g., Trolox or ascorbic acid) and report results as IC₅₀ or mg Trolox equivalent/g for comparability. 10. Antimicrobial claims cannot use “broad-spectrum” without testing a representative panel. 11. Key MAE factors were omitted (type of solvent, solvent concentration, power, temperature, time, solid:liquid ratio). If you want to claim optimization, perform a DOE (factorial or RSM). Otherwise clearly present the work as a screening study. 12. Rephrase or remove any absolute claims (e.g., “broad-spectrum”, “no degradation”) unless supported by data. 13. Reword the heat-cycling sentence: heating–cooling is a preliminary shelf-stress test, not a full simulation of long-term stability. Use careful language. 14. Update and expand references: several cited works are dated — include recent MAE and cosmeceutical formulation literature. 15. Drying methods, whether heat-based (e.g., tray drying) or non-thermal (e.g., freeze-drying), are well-documented for their influence on phenolic compound stability and retention. Please clarify the rationale for selecting this factor as the focus of investigation rather than prioritizing the optimization of parameters directly affecting the extraction process. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? No Are sufficient details of methods and analysis provided to allow replication by others? No If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Partly Are the conclusions drawn adequately supported by the results? No Competing Interests: No competing interests were disclosed. Reviewer Expertise: Non-conventional extraction. Bioactive compounds, Cosmetics, Cell-culture assays, Formulations I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Khat-Udomkiri N. Reviewer Report For: Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Techniques and Its Value Addition in Cosmeceutical Products [version 1; peer review: 1 approved, 1 not approved] . F1000Research 2025, 14 :796 ( https://doi.org/10.5256/f1000research.181624.r413222 ) The direct URL for this report is: https://f1000research.com/articles/14-796/v1#referee-response-413222 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 13 Oct 2025 Doungporn Amornlerdpison , Center of Excellence in Agricultural Innovation for Graduate Entrepreneur, Maejo University, Nong Han, Thailand 13 Oct 2025 Author Response Reviewer 2 (Not Approved) 1. You must cite and discuss prior MAE work on P. amboinicus (e.g., the paper you identified: DOI: 10.3303/CET1972067). Clearly state how your study differs ... Continue reading Reviewer 2 (Not Approved) 1. You must cite and discuss prior MAE work on P. amboinicus (e.g., the paper you identified: DOI: 10.3303/CET1972067). Clearly state how your study differs (new endpoints, different solvent system, biological assays, formulation work, scale-up considerations, etc.). Right now the manuscript reads like a repeat of existing work. Response : Our study differs by evaluating drying methods (tray vs freeze-drying), biological activities (antimicrobial and anti-inflammatory properties) and formulation into a cream with stability testing. 2. Do not claim you “optimized” extraction conditions unless a formal design of experiments (e.g., RSM, factorial) or an explicit optimization protocol was applied. If you did not run an optimization design, rephrase to “screened”, “evaluated” or “investigated”. Response : We agree and have rephrased “optimized” to “evaluated”, Page 2 unless clearly supported by data. 3. Temperature is critical for MAE outcomes and must be reported for every condition. Report temperature profiles for all MAE runs. Response: We have added the extraction temperature (40 °C under 450 W microwave power) in the Methods section, Page 3 4. Explain the basis for selecting caffeic acid as the principal marker. Was it the most abundant compound in the extract, the most bioactive, or selected for analytical convenience? Response : We selected caffeic acid as a marker because it was the most abundant phenolic identified in preliminary HPLC profiling. This justification has been added, Page 3. 5. Natural extracts are mixtures — identify and quantify multiple major phenolics, not only one. Provide HPLC chromatograms and quantification for at least the top 3–5 phenolic compounds. Response: In this study, a representative phenolic compound, caffeic acid was selected as a biomarker for quantification due to its well-documented bioactivity. Furthermore, the use of total phenolic content (TPC) provides a reliable estimation of the overall phenolic load in the extracts. This approach enables holistic evaluation of the extract's functionality while minimizing the complexity, time, and cost associated with full chromatographic profiling. Future work may include expanded HPLC profiling to better understand the full phenolic spectrum and assess potential synergistic effects among individual compounds. 6. Provide a full cream formulation (all ingredients, % w/w), the exact amount of extract incorporated, and the rationale for the selected concentration(s). Response: We have revised the section to provide the complete formulation with all ingredients and concentrations (% w/w) The formulation was based on the minimum inhibitory concentration (MIC) of PF15, determined to be 7.8 mg/mL against Staphylococcus aureus . For a 100 g cream formulation (equivalent to approximately 100 mL), the required amount of extract was calculated as 780 mg, corresponding to 0.78% (w/w) of the total formulation. The 0.8% concentration was selected to ensure antimicrobial efficacy while maintaining formulation stability, Page 5-6. 7. You state extract was included during emulsification: explain and justify this choice. Phenolic compounds is heat-sensitive; if used in a heated phase, show data proving it remains active after processing. Response: The PF15 extract was incorporated during the emulsification step to ensure uniform distribution of the bioactive compounds within the cream matrix. Although phenolic compounds are generally considered heat-sensitive, our results demonstrate that the antimicrobial activity of the extract was largely retained after processing. The PF15 extract alone showed inhibition zones of 13–15 mm (Table 3, Page 12) against selected skin pathogens, while the formulated PF cream exhibited inhibition zones of 11–12 mm (Page 16). The slight reduction indicates minor thermal impact; however, the preserved inhibitory activity confirms that the phenolic constituents remained bioactive after emulsification. 8. You cannot assert “no degradation” based on visual inspection alone. Provide objective stability data: HPLC profiles (initial vs after processing and after stability cycles), TPC, and at least one chemical marker quantified before/after heating and after storage cycles. Replace subjective color assessments with ΔE measurements and report ΔE for color change. Response: Deleted no degradation. We acknowledge the reviewer’s comment regarding the need for objective stability data. In our study, instead of quantifying individual phenolic compounds within the cream formulation, stability was evaluated based on the functional antimicrobial activity of the product. This approach was selected because the cream contains multiple ingredients. Therefore, antimicrobial efficacy against target skin pathogens was used as a practical stability indicator (Page 16), confirming that the bioactive effect of PF15 was retained after processing and throughout the stability cycles. 9. DPPH must include a positive control (e.g., Trolox or ascorbic acid) and report results as IC₅₀ or mg Trolox equivalent/g for comparability. Response : We used gallic acid as the positive control, consistent with its use in the phenolic content determination. Figure 7 has been updated to include gallic acid as the control. 10. Antimicrobial claims cannot use “broad-spectrum” without testing a representative panel. Response : Deleted broad-spectrum from conclusion. 11. Key MAE factors were omitted (type of solvent, solvent concentration, power, temperature, time, solid:liquid ratio). If you want to claim optimization, perform a DOE (factorial or RSM). Otherwise clearly present the work as a screening study. Response : We have now reported solvent type and concentration (50% ethanol), power (450 W), temperature (40 °C), extraction time (10–20 min), and solid–liquid ratio (1:10 v/v) in the Methods section, Page 3. 12. Rephrase or remove any absolute claims (e.g., “broad-spectrum”, “no degradation”) unless supported by data. Response : Statements such as “no degradation” and “broad-spectrum” have been rephrased to reflect the data accurately. 13. Reword the heat-cycling sentence: heating–cooling is a preliminary shelf-stress test, not a full simulation of long-term stability. Use careful language. Response : We have reworded this as a “preliminary stress test to evaluate its stability under accelerated conditions”, Page 16 14. Update and expand references: several cited works are dated — include recent MAE and cosmeceutical formulation literature. Response : We have updated the reference with recent MAE and other green extraction, Page 17-18 References updated: [43] 15. Drying methods, whether heat-based (e.g., tray drying) or non-thermal (e.g., freeze-drying), are well-documented for their influence on phenolic compound stability and retention. Please clarify the rationale for selecting this factor as the focus of investigation rather than prioritizing the optimization of parameters directly affecting the extraction process. Response : Our unique focus on drying and extraction methods relevance for industrial-scale processing and use and effective cost for commercial. We believe these revisions strengthen the manuscript significantly by clarifying novelty, improving methodological transparency, and ensuring accurate claims. We hope that the revised version now satisfactorily addresses all reviewer concerns and will be considered suitable for approval Reviewer 2 (Not Approved) 1. You must cite and discuss prior MAE work on P. amboinicus (e.g., the paper you identified: DOI: 10.3303/CET1972067). Clearly state how your study differs (new endpoints, different solvent system, biological assays, formulation work, scale-up considerations, etc.). Right now the manuscript reads like a repeat of existing work. Response : Our study differs by evaluating drying methods (tray vs freeze-drying), biological activities (antimicrobial and anti-inflammatory properties) and formulation into a cream with stability testing. 2. Do not claim you “optimized” extraction conditions unless a formal design of experiments (e.g., RSM, factorial) or an explicit optimization protocol was applied. If you did not run an optimization design, rephrase to “screened”, “evaluated” or “investigated”. Response : We agree and have rephrased “optimized” to “evaluated”, Page 2 unless clearly supported by data. 3. Temperature is critical for MAE outcomes and must be reported for every condition. Report temperature profiles for all MAE runs. Response: We have added the extraction temperature (40 °C under 450 W microwave power) in the Methods section, Page 3 4. Explain the basis for selecting caffeic acid as the principal marker. Was it the most abundant compound in the extract, the most bioactive, or selected for analytical convenience? Response : We selected caffeic acid as a marker because it was the most abundant phenolic identified in preliminary HPLC profiling. This justification has been added, Page 3. 5. Natural extracts are mixtures — identify and quantify multiple major phenolics, not only one. Provide HPLC chromatograms and quantification for at least the top 3–5 phenolic compounds. Response: In this study, a representative phenolic compound, caffeic acid was selected as a biomarker for quantification due to its well-documented bioactivity. Furthermore, the use of total phenolic content (TPC) provides a reliable estimation of the overall phenolic load in the extracts. This approach enables holistic evaluation of the extract's functionality while minimizing the complexity, time, and cost associated with full chromatographic profiling. Future work may include expanded HPLC profiling to better understand the full phenolic spectrum and assess potential synergistic effects among individual compounds. 6. Provide a full cream formulation (all ingredients, % w/w), the exact amount of extract incorporated, and the rationale for the selected concentration(s). Response: We have revised the section to provide the complete formulation with all ingredients and concentrations (% w/w) The formulation was based on the minimum inhibitory concentration (MIC) of PF15, determined to be 7.8 mg/mL against Staphylococcus aureus . For a 100 g cream formulation (equivalent to approximately 100 mL), the required amount of extract was calculated as 780 mg, corresponding to 0.78% (w/w) of the total formulation. The 0.8% concentration was selected to ensure antimicrobial efficacy while maintaining formulation stability, Page 5-6. 7. You state extract was included during emulsification: explain and justify this choice. Phenolic compounds is heat-sensitive; if used in a heated phase, show data proving it remains active after processing. Response: The PF15 extract was incorporated during the emulsification step to ensure uniform distribution of the bioactive compounds within the cream matrix. Although phenolic compounds are generally considered heat-sensitive, our results demonstrate that the antimicrobial activity of the extract was largely retained after processing. The PF15 extract alone showed inhibition zones of 13–15 mm (Table 3, Page 12) against selected skin pathogens, while the formulated PF cream exhibited inhibition zones of 11–12 mm (Page 16). The slight reduction indicates minor thermal impact; however, the preserved inhibitory activity confirms that the phenolic constituents remained bioactive after emulsification. 8. You cannot assert “no degradation” based on visual inspection alone. Provide objective stability data: HPLC profiles (initial vs after processing and after stability cycles), TPC, and at least one chemical marker quantified before/after heating and after storage cycles. Replace subjective color assessments with ΔE measurements and report ΔE for color change. Response: Deleted no degradation. We acknowledge the reviewer’s comment regarding the need for objective stability data. In our study, instead of quantifying individual phenolic compounds within the cream formulation, stability was evaluated based on the functional antimicrobial activity of the product. This approach was selected because the cream contains multiple ingredients. Therefore, antimicrobial efficacy against target skin pathogens was used as a practical stability indicator (Page 16), confirming that the bioactive effect of PF15 was retained after processing and throughout the stability cycles. 9. DPPH must include a positive control (e.g., Trolox or ascorbic acid) and report results as IC₅₀ or mg Trolox equivalent/g for comparability. Response : We used gallic acid as the positive control, consistent with its use in the phenolic content determination. Figure 7 has been updated to include gallic acid as the control. 10. Antimicrobial claims cannot use “broad-spectrum” without testing a representative panel. Response : Deleted broad-spectrum from conclusion. 11. Key MAE factors were omitted (type of solvent, solvent concentration, power, temperature, time, solid:liquid ratio). If you want to claim optimization, perform a DOE (factorial or RSM). Otherwise clearly present the work as a screening study. Response : We have now reported solvent type and concentration (50% ethanol), power (450 W), temperature (40 °C), extraction time (10–20 min), and solid–liquid ratio (1:10 v/v) in the Methods section, Page 3. 12. Rephrase or remove any absolute claims (e.g., “broad-spectrum”, “no degradation”) unless supported by data. Response : Statements such as “no degradation” and “broad-spectrum” have been rephrased to reflect the data accurately. 13. Reword the heat-cycling sentence: heating–cooling is a preliminary shelf-stress test, not a full simulation of long-term stability. Use careful language. Response : We have reworded this as a “preliminary stress test to evaluate its stability under accelerated conditions”, Page 16 14. Update and expand references: several cited works are dated — include recent MAE and cosmeceutical formulation literature. Response : We have updated the reference with recent MAE and other green extraction, Page 17-18 References updated: [43] 15. Drying methods, whether heat-based (e.g., tray drying) or non-thermal (e.g., freeze-drying), are well-documented for their influence on phenolic compound stability and retention. Please clarify the rationale for selecting this factor as the focus of investigation rather than prioritizing the optimization of parameters directly affecting the extraction process. Response : Our unique focus on drying and extraction methods relevance for industrial-scale processing and use and effective cost for commercial. We believe these revisions strengthen the manuscript significantly by clarifying novelty, improving methodological transparency, and ensuring accurate claims. We hope that the revised version now satisfactorily addresses all reviewer concerns and will be considered suitable for approval Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 13 Oct 2025 Doungporn Amornlerdpison , Center of Excellence in Agricultural Innovation for Graduate Entrepreneur, Maejo University, Nong Han, Thailand 13 Oct 2025 Author Response Reviewer 2 (Not Approved) 1. You must cite and discuss prior MAE work on P. amboinicus (e.g., the paper you identified: DOI: 10.3303/CET1972067). Clearly state how your study differs ... Continue reading Reviewer 2 (Not Approved) 1. You must cite and discuss prior MAE work on P. amboinicus (e.g., the paper you identified: DOI: 10.3303/CET1972067). Clearly state how your study differs (new endpoints, different solvent system, biological assays, formulation work, scale-up considerations, etc.). Right now the manuscript reads like a repeat of existing work. Response : Our study differs by evaluating drying methods (tray vs freeze-drying), biological activities (antimicrobial and anti-inflammatory properties) and formulation into a cream with stability testing. 2. Do not claim you “optimized” extraction conditions unless a formal design of experiments (e.g., RSM, factorial) or an explicit optimization protocol was applied. If you did not run an optimization design, rephrase to “screened”, “evaluated” or “investigated”. Response : We agree and have rephrased “optimized” to “evaluated”, Page 2 unless clearly supported by data. 3. Temperature is critical for MAE outcomes and must be reported for every condition. Report temperature profiles for all MAE runs. Response: We have added the extraction temperature (40 °C under 450 W microwave power) in the Methods section, Page 3 4. Explain the basis for selecting caffeic acid as the principal marker. Was it the most abundant compound in the extract, the most bioactive, or selected for analytical convenience? Response : We selected caffeic acid as a marker because it was the most abundant phenolic identified in preliminary HPLC profiling. This justification has been added, Page 3. 5. Natural extracts are mixtures — identify and quantify multiple major phenolics, not only one. Provide HPLC chromatograms and quantification for at least the top 3–5 phenolic compounds. Response: In this study, a representative phenolic compound, caffeic acid was selected as a biomarker for quantification due to its well-documented bioactivity. Furthermore, the use of total phenolic content (TPC) provides a reliable estimation of the overall phenolic load in the extracts. This approach enables holistic evaluation of the extract's functionality while minimizing the complexity, time, and cost associated with full chromatographic profiling. Future work may include expanded HPLC profiling to better understand the full phenolic spectrum and assess potential synergistic effects among individual compounds. 6. Provide a full cream formulation (all ingredients, % w/w), the exact amount of extract incorporated, and the rationale for the selected concentration(s). Response: We have revised the section to provide the complete formulation with all ingredients and concentrations (% w/w) The formulation was based on the minimum inhibitory concentration (MIC) of PF15, determined to be 7.8 mg/mL against Staphylococcus aureus . For a 100 g cream formulation (equivalent to approximately 100 mL), the required amount of extract was calculated as 780 mg, corresponding to 0.78% (w/w) of the total formulation. The 0.8% concentration was selected to ensure antimicrobial efficacy while maintaining formulation stability, Page 5-6. 7. You state extract was included during emulsification: explain and justify this choice. Phenolic compounds is heat-sensitive; if used in a heated phase, show data proving it remains active after processing. Response: The PF15 extract was incorporated during the emulsification step to ensure uniform distribution of the bioactive compounds within the cream matrix. Although phenolic compounds are generally considered heat-sensitive, our results demonstrate that the antimicrobial activity of the extract was largely retained after processing. The PF15 extract alone showed inhibition zones of 13–15 mm (Table 3, Page 12) against selected skin pathogens, while the formulated PF cream exhibited inhibition zones of 11–12 mm (Page 16). The slight reduction indicates minor thermal impact; however, the preserved inhibitory activity confirms that the phenolic constituents remained bioactive after emulsification. 8. You cannot assert “no degradation” based on visual inspection alone. Provide objective stability data: HPLC profiles (initial vs after processing and after stability cycles), TPC, and at least one chemical marker quantified before/after heating and after storage cycles. Replace subjective color assessments with ΔE measurements and report ΔE for color change. Response: Deleted no degradation. We acknowledge the reviewer’s comment regarding the need for objective stability data. In our study, instead of quantifying individual phenolic compounds within the cream formulation, stability was evaluated based on the functional antimicrobial activity of the product. This approach was selected because the cream contains multiple ingredients. Therefore, antimicrobial efficacy against target skin pathogens was used as a practical stability indicator (Page 16), confirming that the bioactive effect of PF15 was retained after processing and throughout the stability cycles. 9. DPPH must include a positive control (e.g., Trolox or ascorbic acid) and report results as IC₅₀ or mg Trolox equivalent/g for comparability. Response : We used gallic acid as the positive control, consistent with its use in the phenolic content determination. Figure 7 has been updated to include gallic acid as the control. 10. Antimicrobial claims cannot use “broad-spectrum” without testing a representative panel. Response : Deleted broad-spectrum from conclusion. 11. Key MAE factors were omitted (type of solvent, solvent concentration, power, temperature, time, solid:liquid ratio). If you want to claim optimization, perform a DOE (factorial or RSM). Otherwise clearly present the work as a screening study. Response : We have now reported solvent type and concentration (50% ethanol), power (450 W), temperature (40 °C), extraction time (10–20 min), and solid–liquid ratio (1:10 v/v) in the Methods section, Page 3. 12. Rephrase or remove any absolute claims (e.g., “broad-spectrum”, “no degradation”) unless supported by data. Response : Statements such as “no degradation” and “broad-spectrum” have been rephrased to reflect the data accurately. 13. Reword the heat-cycling sentence: heating–cooling is a preliminary shelf-stress test, not a full simulation of long-term stability. Use careful language. Response : We have reworded this as a “preliminary stress test to evaluate its stability under accelerated conditions”, Page 16 14. Update and expand references: several cited works are dated — include recent MAE and cosmeceutical formulation literature. Response : We have updated the reference with recent MAE and other green extraction, Page 17-18 References updated: [43] 15. Drying methods, whether heat-based (e.g., tray drying) or non-thermal (e.g., freeze-drying), are well-documented for their influence on phenolic compound stability and retention. Please clarify the rationale for selecting this factor as the focus of investigation rather than prioritizing the optimization of parameters directly affecting the extraction process. Response : Our unique focus on drying and extraction methods relevance for industrial-scale processing and use and effective cost for commercial. We believe these revisions strengthen the manuscript significantly by clarifying novelty, improving methodological transparency, and ensuring accurate claims. We hope that the revised version now satisfactorily addresses all reviewer concerns and will be considered suitable for approval Reviewer 2 (Not Approved) 1. You must cite and discuss prior MAE work on P. amboinicus (e.g., the paper you identified: DOI: 10.3303/CET1972067). Clearly state how your study differs (new endpoints, different solvent system, biological assays, formulation work, scale-up considerations, etc.). Right now the manuscript reads like a repeat of existing work. Response : Our study differs by evaluating drying methods (tray vs freeze-drying), biological activities (antimicrobial and anti-inflammatory properties) and formulation into a cream with stability testing. 2. Do not claim you “optimized” extraction conditions unless a formal design of experiments (e.g., RSM, factorial) or an explicit optimization protocol was applied. If you did not run an optimization design, rephrase to “screened”, “evaluated” or “investigated”. Response : We agree and have rephrased “optimized” to “evaluated”, Page 2 unless clearly supported by data. 3. Temperature is critical for MAE outcomes and must be reported for every condition. Report temperature profiles for all MAE runs. Response: We have added the extraction temperature (40 °C under 450 W microwave power) in the Methods section, Page 3 4. Explain the basis for selecting caffeic acid as the principal marker. Was it the most abundant compound in the extract, the most bioactive, or selected for analytical convenience? Response : We selected caffeic acid as a marker because it was the most abundant phenolic identified in preliminary HPLC profiling. This justification has been added, Page 3. 5. Natural extracts are mixtures — identify and quantify multiple major phenolics, not only one. Provide HPLC chromatograms and quantification for at least the top 3–5 phenolic compounds. Response: In this study, a representative phenolic compound, caffeic acid was selected as a biomarker for quantification due to its well-documented bioactivity. Furthermore, the use of total phenolic content (TPC) provides a reliable estimation of the overall phenolic load in the extracts. This approach enables holistic evaluation of the extract's functionality while minimizing the complexity, time, and cost associated with full chromatographic profiling. Future work may include expanded HPLC profiling to better understand the full phenolic spectrum and assess potential synergistic effects among individual compounds. 6. Provide a full cream formulation (all ingredients, % w/w), the exact amount of extract incorporated, and the rationale for the selected concentration(s). Response: We have revised the section to provide the complete formulation with all ingredients and concentrations (% w/w) The formulation was based on the minimum inhibitory concentration (MIC) of PF15, determined to be 7.8 mg/mL against Staphylococcus aureus . For a 100 g cream formulation (equivalent to approximately 100 mL), the required amount of extract was calculated as 780 mg, corresponding to 0.78% (w/w) of the total formulation. The 0.8% concentration was selected to ensure antimicrobial efficacy while maintaining formulation stability, Page 5-6. 7. You state extract was included during emulsification: explain and justify this choice. Phenolic compounds is heat-sensitive; if used in a heated phase, show data proving it remains active after processing. Response: The PF15 extract was incorporated during the emulsification step to ensure uniform distribution of the bioactive compounds within the cream matrix. Although phenolic compounds are generally considered heat-sensitive, our results demonstrate that the antimicrobial activity of the extract was largely retained after processing. The PF15 extract alone showed inhibition zones of 13–15 mm (Table 3, Page 12) against selected skin pathogens, while the formulated PF cream exhibited inhibition zones of 11–12 mm (Page 16). The slight reduction indicates minor thermal impact; however, the preserved inhibitory activity confirms that the phenolic constituents remained bioactive after emulsification. 8. You cannot assert “no degradation” based on visual inspection alone. Provide objective stability data: HPLC profiles (initial vs after processing and after stability cycles), TPC, and at least one chemical marker quantified before/after heating and after storage cycles. Replace subjective color assessments with ΔE measurements and report ΔE for color change. Response: Deleted no degradation. We acknowledge the reviewer’s comment regarding the need for objective stability data. In our study, instead of quantifying individual phenolic compounds within the cream formulation, stability was evaluated based on the functional antimicrobial activity of the product. This approach was selected because the cream contains multiple ingredients. Therefore, antimicrobial efficacy against target skin pathogens was used as a practical stability indicator (Page 16), confirming that the bioactive effect of PF15 was retained after processing and throughout the stability cycles. 9. DPPH must include a positive control (e.g., Trolox or ascorbic acid) and report results as IC₅₀ or mg Trolox equivalent/g for comparability. Response : We used gallic acid as the positive control, consistent with its use in the phenolic content determination. Figure 7 has been updated to include gallic acid as the control. 10. Antimicrobial claims cannot use “broad-spectrum” without testing a representative panel. Response : Deleted broad-spectrum from conclusion. 11. Key MAE factors were omitted (type of solvent, solvent concentration, power, temperature, time, solid:liquid ratio). If you want to claim optimization, perform a DOE (factorial or RSM). Otherwise clearly present the work as a screening study. Response : We have now reported solvent type and concentration (50% ethanol), power (450 W), temperature (40 °C), extraction time (10–20 min), and solid–liquid ratio (1:10 v/v) in the Methods section, Page 3. 12. Rephrase or remove any absolute claims (e.g., “broad-spectrum”, “no degradation”) unless supported by data. Response : Statements such as “no degradation” and “broad-spectrum” have been rephrased to reflect the data accurately. 13. Reword the heat-cycling sentence: heating–cooling is a preliminary shelf-stress test, not a full simulation of long-term stability. Use careful language. Response : We have reworded this as a “preliminary stress test to evaluate its stability under accelerated conditions”, Page 16 14. Update and expand references: several cited works are dated — include recent MAE and cosmeceutical formulation literature. Response : We have updated the reference with recent MAE and other green extraction, Page 17-18 References updated: [43] 15. Drying methods, whether heat-based (e.g., tray drying) or non-thermal (e.g., freeze-drying), are well-documented for their influence on phenolic compound stability and retention. Please clarify the rationale for selecting this factor as the focus of investigation rather than prioritizing the optimization of parameters directly affecting the extraction process. Response : Our unique focus on drying and extraction methods relevance for industrial-scale processing and use and effective cost for commercial. We believe these revisions strengthen the manuscript significantly by clarifying novelty, improving methodological transparency, and ensuring accurate claims. We hope that the revised version now satisfactorily addresses all reviewer concerns and will be considered suitable for approval Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Close Report a concern COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Srivilai J. Reviewer Report For: Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Techniques and Its Value Addition in Cosmeceutical Products [version 1; peer review: 1 approved, 1 not approved] . F1000Research 2025, 14 :796 ( https://doi.org/10.5256/f1000research.181624.r407511 ) The direct URL for this report is: https://f1000research.com/articles/14-796/v1#referee-response-407511 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 15 Sep 2025 Jukkarin Srivilai , University of Phayao, Phayao, Thailand Approved VIEWS 0 https://doi.org/10.5256/f1000research.181624.r407511 The manuscript titled “Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Technique and Its Value Addition in Cosmeceutical Products” presents interesting and technically relevant work. However, several issues need to be addressed before indexing. The following comments are provided for the authors’ ... Continue reading READ ALL The manuscript titled “Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Technique and Its Value Addition in Cosmeceutical Products” presents interesting and technically relevant work. However, several issues need to be addressed before indexing. The following comments are provided for the authors’ consideration: Title: The word “Techniques” should be singular (“Technique”). Abstract: In the Methods section, the last sentence should be revised to: “Anti-inflammatory potential of PF15 ….” Dosage form: The authors should clearly state the dosage form of the cream throughout the manuscript, or at least specify somewhere that it is an emulsion type. Introduction: In the third paragraph, please verify reference [6] and revise the sentence for better grammatical clarity. Materials and Methods: The description of the 50% ethanol extraction should specify the solvent used (e.g., DI water or another solvent). Chemical analysis (HPLC): The methodology should clarify whether standard compounds were used to construct the calibration curve for determining the relative amount of caffeic acid, or if another approach was applied. Please provide details. Table 1: The column “Mode” should be labeled “Mode of elution.” Abbreviations: Ensure that abbreviations follow the full name consistently (e.g., Minimal Inhibitory Concentration (MIC)). Please check throughout the manuscript. Cream formulation with PF15: Provide more details on the formulation ratio, type of emulsion, and preparation technique. Results and Discussion: Verify the unit of yield, as it may be incorrect. In Figure 3, the retention time of the caffeic acid standard does not match the retention time mentioned in the Results and Discussion section. Please correct this, as it is a critical point for analysis. Comparative discussion: Expand the discussion to include other extraction techniques or green solvents. You may cite the following articles: [Reference 1] [Reference 2] Scientific names: On page 10, in the antioxidant activity section, the correct scientific name is P. amboinicus . Similarly, check all bacterial names and ensure consistency throughout the manuscript. Table 2: Use a big letter instead of “inhibition” to be Inhibition. The caption should specify the sample/disc amount (e.g., 5 mg/disc). Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes References 1. Saesue K, Matwangsang S, Rungsang T, Khorana N, et al.: Deep eutectic-based microemulsion: A multifunctional system for enhancing extraction efficiency, stability and antityrosinase, antioxidant activities of oxyresveratrol from Artocarpus lakoocha Roxb. Journal of Molecular Liquids . 2025; 436 . Publisher Full Text 2. Rungsang T, Aimjongjun S, Aoonboontum P, Matwangsang S, et al.: A novel deep eutectic and eutectic-based microemulsion systems for enhanced extraction, stabilization and antioxidant activity of artocarpin, isocyclomorusin, and cycloartocarpin from agricultural byproduct of Artocarpus heterophyllus Lam. Microchemical Journal . 2025; 217 . Publisher Full Text Competing Interests: No competing interests were disclosed. Reviewer Expertise: Plant extraction technologies I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Srivilai J. Reviewer Report For: Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Techniques and Its Value Addition in Cosmeceutical Products [version 1; peer review: 1 approved, 1 not approved] . F1000Research 2025, 14 :796 ( https://doi.org/10.5256/f1000research.181624.r407511 ) The direct URL for this report is: https://f1000research.com/articles/14-796/v1#referee-response-407511 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 13 Oct 2025 Doungporn Amornlerdpison , Center of Excellence in Agricultural Innovation for Graduate Entrepreneur, Maejo University, Nong Han, Thailand 13 Oct 2025 Author Response We would like to sincerely thank the reviewers for the valuable and constructive comments. We have carefully revised the manuscript according to all suggestions, and below we provide a point-by-point ... Continue reading We would like to sincerely thank the reviewers for the valuable and constructive comments. We have carefully revised the manuscript according to all suggestions, and below we provide a point-by-point response. The changes have been made as highlighted in “red font” in revised manuscript. We hope our revision has improved the paper to a satisfactory level. Reviewer 1 (Approved) 1. Title: The word “Techniques” should be singular (“Technique”). Corrected to “Technique.” 2. Abstract: In the Methods section, the last sentence should be revised to: “Anti-inflammatory potential of PF15 ….” Revised: “Anti-inflammatory potential of PF15 was evaluated in LPS-stimulated THP-1 macrophages.” 3.Dosage form: The authors should clearly state the dosage form of the cream throughout the manuscript, or at least specify somewhere that it is an emulsion type. Stated in Part: Product Development and Evaluation, Page 5 “The extract was incorporated into the cream formulation at a concentration of 0.8%, ensuring antimicrobial efficacy. The aqueous phase containing PF15 was heated to 70 °C, combined with the oil phase, and emulsified for 30 minutes. The cream was then cooled to 35–40 °C, mixed until homogeneous, and stored for further analysis. A topical cream was developed by incorporating the PF15 extract into a water-in-oil emulsion system. The detailed composition of the cream is presented in Table 2.” 4.Introduction: In the third paragraph, please verify reference [6] and revise the sentence for better grammatical clarity. Verified and corrected sentence, Page 2. “A recent study demonstrated the antimicrobial efficacy of P. amboinicus leaf extract against pathogens such as Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, and the acne-associated bacterium Cutibacterium acnes, thereby reinforcing its potential application in cosmetic formulations. 6 5.Materials and Methods: The description of the 50% ethanol extraction should specify the solvent used (e.g., DI water or another solvent). Clarified as ethanol + deionized water, Page 3. “The 50% ethanol solution was prepared using a 1:1 mixture of ethanol and deionized water.” - Chemical analysis (HPLC): The methodology should clarify whether standard compounds were used to construct the calibration curve for determining the relative amount of caffeic acid, or if another approach was applied. Please provide details. Clarified as “The standard of caffeic acid was used to construct the calibration curve.” - Table 1: The column “Mode” should be labeled “Mode of elution.” Corrected. Mode of elution - Abbreviations: Ensure that abbreviations follow the full name consistently (e.g., Minimal Inhibitory Concentration (MIC)). Please check throughout the manuscript. Checked and corrected; Minimal Inhibitory Concentration (MIC) - Cream formulation with PF15: Provide more details on the formulation ratio, type of emulsion, and preparation technique. Full formulation ratios, emulsion type, and preparation technique added, Page 5-6. 6.Results and Discussion: - Verify the unit of yield, as it may be incorrect. Corrected to % w/w dry weight - Figure 3, the retention time of the caffeic acid standard does not match the retention time mentioned in the Results and Discussion section. Please correct this, as it is a critical point for analysis. Corrected to 7.8 min retention time for caffeic acid. 7.Comparative discussion: Expand the discussion to include other extraction techniques or green solvents. You may cite the following articles: [Reference 1] [Reference 2] Expanded with ultrasound-assisted, pressurized liquid, and other green solvent extraction, Page 17-18 References updated: [43] We would like to sincerely thank the reviewers for the valuable and constructive comments. We have carefully revised the manuscript according to all suggestions, and below we provide a point-by-point response. The changes have been made as highlighted in “red font” in revised manuscript. We hope our revision has improved the paper to a satisfactory level. Reviewer 1 (Approved) 1. Title: The word “Techniques” should be singular (“Technique”). Corrected to “Technique.” 2. Abstract: In the Methods section, the last sentence should be revised to: “Anti-inflammatory potential of PF15 ….” Revised: “Anti-inflammatory potential of PF15 was evaluated in LPS-stimulated THP-1 macrophages.” 3.Dosage form: The authors should clearly state the dosage form of the cream throughout the manuscript, or at least specify somewhere that it is an emulsion type. Stated in Part: Product Development and Evaluation, Page 5 “The extract was incorporated into the cream formulation at a concentration of 0.8%, ensuring antimicrobial efficacy. The aqueous phase containing PF15 was heated to 70 °C, combined with the oil phase, and emulsified for 30 minutes. The cream was then cooled to 35–40 °C, mixed until homogeneous, and stored for further analysis. A topical cream was developed by incorporating the PF15 extract into a water-in-oil emulsion system. The detailed composition of the cream is presented in Table 2.” 4.Introduction: In the third paragraph, please verify reference [6] and revise the sentence for better grammatical clarity. Verified and corrected sentence, Page 2. “A recent study demonstrated the antimicrobial efficacy of P. amboinicus leaf extract against pathogens such as Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, and the acne-associated bacterium Cutibacterium acnes, thereby reinforcing its potential application in cosmetic formulations. 6 5.Materials and Methods: The description of the 50% ethanol extraction should specify the solvent used (e.g., DI water or another solvent). Clarified as ethanol + deionized water, Page 3. “The 50% ethanol solution was prepared using a 1:1 mixture of ethanol and deionized water.” - Chemical analysis (HPLC): The methodology should clarify whether standard compounds were used to construct the calibration curve for determining the relative amount of caffeic acid, or if another approach was applied. Please provide details. Clarified as “The standard of caffeic acid was used to construct the calibration curve.” - Table 1: The column “Mode” should be labeled “Mode of elution.” Corrected. Mode of elution - Abbreviations: Ensure that abbreviations follow the full name consistently (e.g., Minimal Inhibitory Concentration (MIC)). Please check throughout the manuscript. Checked and corrected; Minimal Inhibitory Concentration (MIC) - Cream formulation with PF15: Provide more details on the formulation ratio, type of emulsion, and preparation technique. Full formulation ratios, emulsion type, and preparation technique added, Page 5-6. 6.Results and Discussion: - Verify the unit of yield, as it may be incorrect. Corrected to % w/w dry weight - Figure 3, the retention time of the caffeic acid standard does not match the retention time mentioned in the Results and Discussion section. Please correct this, as it is a critical point for analysis. Corrected to 7.8 min retention time for caffeic acid. 7.Comparative discussion: Expand the discussion to include other extraction techniques or green solvents. You may cite the following articles: [Reference 1] [Reference 2] Expanded with ultrasound-assisted, pressurized liquid, and other green solvent extraction, Page 17-18 References updated: [43] Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 13 Oct 2025 Doungporn Amornlerdpison , Center of Excellence in Agricultural Innovation for Graduate Entrepreneur, Maejo University, Nong Han, Thailand 13 Oct 2025 Author Response We would like to sincerely thank the reviewers for the valuable and constructive comments. We have carefully revised the manuscript according to all suggestions, and below we provide a point-by-point ... Continue reading We would like to sincerely thank the reviewers for the valuable and constructive comments. We have carefully revised the manuscript according to all suggestions, and below we provide a point-by-point response. The changes have been made as highlighted in “red font” in revised manuscript. We hope our revision has improved the paper to a satisfactory level. Reviewer 1 (Approved) 1. Title: The word “Techniques” should be singular (“Technique”). Corrected to “Technique.” 2. Abstract: In the Methods section, the last sentence should be revised to: “Anti-inflammatory potential of PF15 ….” Revised: “Anti-inflammatory potential of PF15 was evaluated in LPS-stimulated THP-1 macrophages.” 3.Dosage form: The authors should clearly state the dosage form of the cream throughout the manuscript, or at least specify somewhere that it is an emulsion type. Stated in Part: Product Development and Evaluation, Page 5 “The extract was incorporated into the cream formulation at a concentration of 0.8%, ensuring antimicrobial efficacy. The aqueous phase containing PF15 was heated to 70 °C, combined with the oil phase, and emulsified for 30 minutes. The cream was then cooled to 35–40 °C, mixed until homogeneous, and stored for further analysis. A topical cream was developed by incorporating the PF15 extract into a water-in-oil emulsion system. The detailed composition of the cream is presented in Table 2.” 4.Introduction: In the third paragraph, please verify reference [6] and revise the sentence for better grammatical clarity. Verified and corrected sentence, Page 2. “A recent study demonstrated the antimicrobial efficacy of P. amboinicus leaf extract against pathogens such as Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, and the acne-associated bacterium Cutibacterium acnes, thereby reinforcing its potential application in cosmetic formulations. 6 5.Materials and Methods: The description of the 50% ethanol extraction should specify the solvent used (e.g., DI water or another solvent). Clarified as ethanol + deionized water, Page 3. “The 50% ethanol solution was prepared using a 1:1 mixture of ethanol and deionized water.” - Chemical analysis (HPLC): The methodology should clarify whether standard compounds were used to construct the calibration curve for determining the relative amount of caffeic acid, or if another approach was applied. Please provide details. Clarified as “The standard of caffeic acid was used to construct the calibration curve.” - Table 1: The column “Mode” should be labeled “Mode of elution.” Corrected. Mode of elution - Abbreviations: Ensure that abbreviations follow the full name consistently (e.g., Minimal Inhibitory Concentration (MIC)). Please check throughout the manuscript. Checked and corrected; Minimal Inhibitory Concentration (MIC) - Cream formulation with PF15: Provide more details on the formulation ratio, type of emulsion, and preparation technique. Full formulation ratios, emulsion type, and preparation technique added, Page 5-6. 6.Results and Discussion: - Verify the unit of yield, as it may be incorrect. Corrected to % w/w dry weight - Figure 3, the retention time of the caffeic acid standard does not match the retention time mentioned in the Results and Discussion section. Please correct this, as it is a critical point for analysis. Corrected to 7.8 min retention time for caffeic acid. 7.Comparative discussion: Expand the discussion to include other extraction techniques or green solvents. You may cite the following articles: [Reference 1] [Reference 2] Expanded with ultrasound-assisted, pressurized liquid, and other green solvent extraction, Page 17-18 References updated: [43] We would like to sincerely thank the reviewers for the valuable and constructive comments. We have carefully revised the manuscript according to all suggestions, and below we provide a point-by-point response. The changes have been made as highlighted in “red font” in revised manuscript. We hope our revision has improved the paper to a satisfactory level. Reviewer 1 (Approved) 1. Title: The word “Techniques” should be singular (“Technique”). Corrected to “Technique.” 2. Abstract: In the Methods section, the last sentence should be revised to: “Anti-inflammatory potential of PF15 ….” Revised: “Anti-inflammatory potential of PF15 was evaluated in LPS-stimulated THP-1 macrophages.” 3.Dosage form: The authors should clearly state the dosage form of the cream throughout the manuscript, or at least specify somewhere that it is an emulsion type. Stated in Part: Product Development and Evaluation, Page 5 “The extract was incorporated into the cream formulation at a concentration of 0.8%, ensuring antimicrobial efficacy. The aqueous phase containing PF15 was heated to 70 °C, combined with the oil phase, and emulsified for 30 minutes. The cream was then cooled to 35–40 °C, mixed until homogeneous, and stored for further analysis. A topical cream was developed by incorporating the PF15 extract into a water-in-oil emulsion system. The detailed composition of the cream is presented in Table 2.” 4.Introduction: In the third paragraph, please verify reference [6] and revise the sentence for better grammatical clarity. Verified and corrected sentence, Page 2. “A recent study demonstrated the antimicrobial efficacy of P. amboinicus leaf extract against pathogens such as Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, and the acne-associated bacterium Cutibacterium acnes, thereby reinforcing its potential application in cosmetic formulations. 6 5.Materials and Methods: The description of the 50% ethanol extraction should specify the solvent used (e.g., DI water or another solvent). Clarified as ethanol + deionized water, Page 3. “The 50% ethanol solution was prepared using a 1:1 mixture of ethanol and deionized water.” - Chemical analysis (HPLC): The methodology should clarify whether standard compounds were used to construct the calibration curve for determining the relative amount of caffeic acid, or if another approach was applied. Please provide details. Clarified as “The standard of caffeic acid was used to construct the calibration curve.” - Table 1: The column “Mode” should be labeled “Mode of elution.” Corrected. Mode of elution - Abbreviations: Ensure that abbreviations follow the full name consistently (e.g., Minimal Inhibitory Concentration (MIC)). Please check throughout the manuscript. Checked and corrected; Minimal Inhibitory Concentration (MIC) - Cream formulation with PF15: Provide more details on the formulation ratio, type of emulsion, and preparation technique. Full formulation ratios, emulsion type, and preparation technique added, Page 5-6. 6.Results and Discussion: - Verify the unit of yield, as it may be incorrect. Corrected to % w/w dry weight - Figure 3, the retention time of the caffeic acid standard does not match the retention time mentioned in the Results and Discussion section. Please correct this, as it is a critical point for analysis. Corrected to 7.8 min retention time for caffeic acid. 7.Comparative discussion: Expand the discussion to include other extraction techniques or green solvents. You may cite the following articles: [Reference 1] [Reference 2] Expanded with ultrasound-assisted, pressurized liquid, and other green solvent extraction, Page 17-18 References updated: [43] Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Close Report a concern COMMENT ON THIS REPORT Comments on this article Comments (0) Version 3 VERSION 3 PUBLISHED 18 Aug 2025 ADD YOUR COMMENT Comment keyboard_arrow_left keyboard_arrow_right Open Peer Review Reviewer Status info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Reviewer Reports Invited Reviewers 1 2 3 4 5 Version 3 (revision) 24 Jan 26 read Version 2 (revision) 13 Oct 25 read read Version 1 18 Aug 25 read read Jukkarin Srivilai , University of Phayao, Phayao, Thailand Nuntawat Khat-Udomkiri , Mae Fah Luang University, Mueang Chiang Rai, Thailand Nguyen Thanh Triet , University of Medicine and Pharmacy at Ho Chi Minh City, Ho Chi Minh City, Vietnam Supenya Chittapun , Thammasat University, Khlong Nueng, Thailand Md Suzauddula Suzauddula , Kansas State University, Manhattan, USA Comments on this article All Comments (0) Add a comment Sign up for content alerts Sign Up You are now signed up to receive this alert Browse by related subjects keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2026 Suzauddula M. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 05 Feb 2026 | for Version 3 Md Suzauddula Suzauddula , Kansas State University, Manhattan, KS, USA 0 Views copyright © 2026 Suzauddula M. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Reviewer Comments Title The title is generally acceptable and reflects the scope of the study. The manuscript contains typographical symbols likely introduced by AI-assisted writing. Please replace the em dash “—” with a hyphen “-” throughout the manuscript. For example: “Two drying techniques—tray drying and freeze-drying—were compared…” should be revised to: “Two drying techniques-tray drying and freeze-drying-were compared…” Introduction Please add appropriate citations to support the statement regarding the adverse effects associated with synthetic chemicals used in cosmetic formulations , including allergic reactions, skin irritation, and potential long-term health risks. Please replace all instances of “—” with “-” in the Introduction section. The following statement requires supporting references: “Plectranthus amboinicus, a perennial herb indigenous to Southeast Asia, has been widely utilized in traditional culinary practices and herbal medicine…” Please add citations supporting its ethnobotanical use, phytochemical composition, and reported antioxidant, antimicrobial, and anti-inflammatory properties . Please revise the writing style to avoid repetitive phrasing. For example, instead of: “A recent study by Ref. 6 demonstrated the antimicrobial efficacy…” use: “A recent study demonstrated the antimicrobial efficacy…” Please avoid unnecessary capitalization within sentences. For example, “Soxhlet” should not be capitalized unless it begins a sentence. The Introduction would benefit from a brief discussion of the enzymes measured in this study , including their biological relevance and importance in cosmeceutical applications . The authors are encouraged to cite recent literature to contextualize their selection. Additionally, please justify the choice of specific microbiota used in this research and explain their relevance to the study objectives. Methods For method validation, please cite the original or reference study from which the compound extraction protocol was adopted or modified. If the chemical characterization method was adapted from previous work, please cite the relevant source and clarify any modifications made. Cytokine measurement by ELISA: Please elaborate on the experimental procedure, including kit details and detection principles, and provide a proper citation (e.g., manufacturer’s protocol or published reference). Cream formulation with PF15: Please validate the formulation method with an appropriate citation from prior literature or formulation guidelines. Please briefly explain the disc diffusion method , including its principle and purpose, with a supporting reference. Table 2 requires justification for the specific percentages of ingredients used . Please explain the rationale for the chosen formulation composition. For microbial enumeration on Dichloran Rose Bengal Chloramphenicol agar, please provide additional methodological details, including: Dilution factors and number of dilution steps Diluent used Incubation temperature and ambient growth conditions Initial microbial load Escherichia coli detection: Please include incubation time, temperature, initial microbial concentration, and a brief explanation of the detection method with citation. Staphylococcus aureus detection: Please include incubation time, temperature, initial microbial concentration, and a brief explanation of the method with citation. Salmonella spp. detection: The absence of dilution requires justification. Please extend the protocol description and include a supporting reference. Please clearly define and classify all formulations/abbreviations used (e.g., PT10, PT15, PT20, PF10, PF15, PF20 ) when first introduced. Results In Figure 2 , no clear significant difference is observable between tray-dried (TD) and freeze-dried (FD) samples at 15 minutes. Please recheck the data and statistical analysis. In Figure 4 , no statistical indicators are shown; however, the authors state that caffeic acid content in freeze-dried extracts was significantly higher. Please clarify: Where the statistical analysis is presented Number of replicates used How significance was determined Figure 5: Please rearrange the bars so that PT10 and PF10 , PT15 and PF15 , and PT20 and PF20 are displayed side by side for easier comparison. The Results section would benefit from the inclusion of recent citations and comparative discussion with similar studies to contextualize the findings. At present, interpretation is limited, and a major revision is recommended to strengthen scientific relevance. Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? No Are sufficient details of methods and analysis provided to allow replication by others? No If applicable, is the statistical analysis and its interpretation appropriate? I cannot comment. A qualified statistician is required. Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Partly Competing Interests No competing interests were disclosed. Reviewer Expertise Bioactive compounds, microbial study, cancer study, and GMO food. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (0) Suzauddula MS. Peer Review Report For: Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Techniques and Its Value Addition in Cosmeceutical Products [version 1; peer review: 1 approved, 1 not approved] . F1000Research 2025, 14 :796 ( https://doi.org/10.5256/f1000research.195380.r443293) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/14-796/v3#referee-response-443293 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2026 Chittapun S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 22 Jan 2026 | for Version 2 Supenya Chittapun , Thammasat University, Khlong Nueng, Thailand 0 Views copyright © 2026 Chittapun S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions The manuscript entitled “Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Technique and Its Value Addition in Cosmeceutical Products” presents an interesting study combining microwave-assisted extraction, biological activity evaluation, and preliminary product formulation. The topic is relevant to the fields of natural products, green extraction technologies, and cosmeceutical development. However, several methodological, reporting, and interpretative issues need to be addressed to improve scientific rigor, reproducibility, and clarity. Many of these points relate to standard requirements for botanical studies, extraction methodology, statistical analysis, and biological interpretation. The following comments are provided section by section, aligned with the manuscript structure, for the authors’ careful consideration. Title and Abstract: -The scientific name of the plant should follow botanical nomenclature rules. At first mention, the full scientific name including the authority should be provided (e.g., Plectranthus amboinicus (Lour.) Spreng.). -Claims made in the abstract regarding extraction efficiency, bioactivity, and formulation performance should be supported by clear methodological descriptions and appropriate statistical analyses in the main text. Introduction: -At the first occurrence, the scientific name of the plant should be written in full with the author citation. Subsequent mentions should use the abbreviated genus name (e.g., P. amboinicus ). If more than one species within the same genus is mentioned, care should be taken to avoid ambiguity. -The statement “Recently, microwave-assisted extraction using 50% ethanol has emerged as a promising alternative” requires a clear and specific citation. Please identify the primary literature supporting this claim. -The introduction would benefit from a clearer justification for focusing on drying methods (tray drying vs. freeze-drying) in addition to extraction time, and how these factors are expected to influence chemical composition and biological activity. Materials and Methods Plant Material and Sample Preparation The source of the plant material should be described in greater detail. Please specify: -Location of collection (area, district, province, country) -Date or season of collection -Plant age or growth stage (if available) -Plant part used -Whether a voucher specimen was prepared and deposited, including voucher number and herbarium This information is essential because phytochemical composition can vary with plant age, season, and geographical location. Extraction and Drying Procedures -The number of experimental replicates should be clearly stated for all extraction experiments, including each extraction time and drying method. -Microwave-assisted extraction may generate heat depending on extraction time and power. Please clarify whether the actual temperature during extraction was monitored and recorded, and if so, how temperature variation was controlled. -Please explain how the absence of residual ethanol in the dried extracts was confirmed prior to biological testing, as residual solvent may influence antimicrobial and cell-based assay results. -Abbreviations such as PT10, PT15, PT20, PF10, PF15, and PF20 should be clearly defined at their first occurrence in the text. Instrumentation and Chemicals -All instruments used should be reported with full details, including brand name, model, manufacturer, and country of origin, to ensure reproducibility. Statistical Analysis - Given that the study involves two independent factors: extraction time and drying method, the use of two-way ANOVA would be more appropriate than one-way ANOVA. This would allow evaluation of the main effect of extraction time, the main effect of drying method and potential interaction effects between these two factors. Microbiological and Cell-Based Assays -Since microbial strains were used in this study, please clarify whether approval or exemption from an institutional biosafety committee was obtained, or explicitly state the biosafety level under which the experiments were conducted. -The rationale for selecting specific extract concentrations for anti-inflammatory assays should be explained, particularly given that cytotoxicity was not observed at much higher concentrations. Results -All figures should include clearly labeled x- and y-axes with appropriate units. -Statistical methods used for data analysis should be stated in each figure legend, including type of statistical test, significance levels, explanation of superscript letters or symbols used to indicate statistical differences. -For Table 3, statistical analysis should be included to support comparisons among treatments. -For Table 4, results should be reported as mean ± standard deviation, based on clearly stated replicate numbers. -For Figure 8, the claim of a dose-dependent effect should be supported by appropriate statistical or trend analysis, rather than descriptive observation alone. -For Figure 9, the authors should expand the discussion to explain the possible biological mechanisms underlying the observed anti-inflammatory effects, rather than only describing changes in cytokine levels. Discussion -As crude extracts were used for biological testing, the authors should discuss which classes of compounds (e.g., caffeic acid, other phenolics, or synergistic mixtures) are most likely responsible for the observed activities. -The discussion should avoid overinterpretation of results and clearly distinguish between experimentally demonstrated effects and proposed mechanisms. Formulation Section -The rationale for selecting a water-in-oil cream base for an anti-acne application should be explained, as oil-rich formulations may influence skin compatibility and acne-related outcomes. Data Presentation and Formatting -Numerical data throughout the manuscript should be reported with consistent decimal places, reflecting the precision of the analytical methods and associated standard deviations. -Please carefully revise the manuscript for language consistency, grammar, and formatting, particularly the use of hyphens and compound terms (e.g., “anti-inflammatory”). In summary, the manuscript addresses a relevant and timely topic; however, substantial revisions are required to improve methodological transparency, statistical rigor, and interpretative depth. Addressing the points outlined above will significantly enhance the scientific quality, reproducibility, and impact of the study. I encourage the authors to carefully revise the manuscript accordingly. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Partly Are the conclusions drawn adequately supported by the results? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise Algal biotechnoloogy I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (0) Chittapun S. Peer Review Report For: Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Techniques and Its Value Addition in Cosmeceutical Products [version 1; peer review: 1 approved, 1 not approved] . F1000Research 2025, 14 :796 ( https://doi.org/10.5256/f1000research.189257.r445681) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/14-796/v2#referee-response-445681 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2026 Triet N. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 14 Jan 2026 | for Version 2 Nguyen Thanh Triet , University of Medicine and Pharmacy at Ho Chi Minh City, Ho Chi Minh City, Vietnam 0 Views copyright © 2026 Triet N. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (1) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions 1. The authors should mention who identified the plant sample? When was the sample collected? What is the name of plant voucher specimen? 2. The authors should mention in detail how they established the optimization of extraction method? 3. The authors need to explain why they used caffeic acid as a marker to evaluate the quality of the extract and optimize the extraction method? 4. Bioactivity assays need to be described in detail, especially which indicators were used to evaluate the results and the calculation formulas. 5. Did the authors see other peaks after 15 minute-run in HPLC chromatogram? Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise pharmacognosy, traditional pharmacy, ethnopharmacology, botanical classification, botanical anatomy, phytochemistry I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (1) Author Response 24 Jan 2026 Doungporn Amornlerdpison, Center of Excellence in Agricultural Innovation for Graduate Entrepreneur, Maejo University, Nong Han, Thailand Response to Reviewer’s Comments We sincerely thank the reviewer for the constructive and insightful comments. Our responses and corresponding revisions are provided below. The changes have been made as highlighted in “red font” in revised manuscript (Version 3). 1. The authors should mention who identified the plant sample? When was the sample collected? What is the name of plant voucher specimen? The plant material ( Plectranthus amboinicus) was taxonomically identified and authenticated by Assoc. Prof. Dr. Yuwalee Unpaprom, a botanist at the Center of Excellence in Agricultural Innovation for Graduate Entrepreneur (Agri Inno), Maejo University, Chiang Mai, Thailand. Fresh leaves were collected in April 2025 from the Maejo area, San Sai District, Chiang Mai Province, Thailand. A voucher specimen has been prepared and deposited at Agri Inno under voucher number Agri Inno- 2574. This information has now been added to the Materials and Methods section of the revised manuscript. 2. The authors should mention in detail how they established the optimization of extraction method? We acknowledge that a formal optimization design, such as Response Surface Methodology or factorial design, was not conducted in this study. Accordingly, the manuscript has been revised to avoid the use of the term “optimized.” Instead, the extraction conditions were systematically evaluated and screened based on microwave-assisted extraction times (5, 10, and 15 minutes) in combination with two drying methods (tray drying and freeze-drying). The selection of the PF15 condition was based on comparative outcomes including extraction yield, total phenolic content, caffeic acid content, and antioxidant activity. The revised manuscript now clearly reflects this screening-based approach. 3. The authors need to explain why they used caffeic acid as a marker to evaluate the quality of the extract and optimize the extraction method? Caffeic acid was selected as a marker compound because it is a well-documented major phenolic constituent of P. amboinicus and is closely associated with its biological activities. Preliminary HPLC screening showed that caffeic acid was the most prominent and consistently detectable peak. Screening with standards of other phenolics, including chlorogenic acid and quercetin, did not show detectable peaks under the applied conditions. Therefore, caffeic acid was used as the representative marker for extract evaluation. 4. Bioactivity assays need to be described in detail, especially which indicators were used to evaluate the results and the calculation formulas. Bioactivity assays are now described in greater detail in the revised manuscript, including the indicators used and calculation approaches. The concentration of PF15 incorporated into the cream formulation was selected based on antimicrobial efficacy and formulation feasibility. The minimum inhibitory concentration (MIC) of PF15 against Staphylococcus aureus was determined to be 7.8 mg/mL . To ensure effective antimicrobial performance in a topical dosage form, PF15 was incorporated at 0.8% w/w , corresponding to approximately 8 mg/g of formulation , consistent with the MIC obtained from in vitro testing. This concentration was selected to ensure sufficient antimicrobial activity while maintaining formulation stability, acceptable sensory properties, and suitability for topical application. Antimicrobial testing of the finished PF15 cream demonstrated inhibition zones of 11–13 mm against selected skin pathogens, confirming that the bioactivity of PF15 was retained after emulsification and thermal processing. Although a slight reduction in inhibition zone diameter compared with the crude extract (13–15 mm) was observed, the extract remained functionally active within the cream matrix. 5. Did the authors see other peaks after 15 minute-run in HPLC chromatogram? As shown in Figures 3 and 4, no additional significant peaks were observed within the 15-minute HPLC run. The chromatographic analysis was designed as a targeted method focusing on the retention time of caffeic acid, which was clearly detected and consistently resolved under the applied analytical conditions. Therefore, the analytical window was limited to 15 minutes to enable reliable and reproducible comparison of extraction conditions using caffeic acid as the representative marker compound. View more View less Competing Interests The authors declare that there are no competing interests regarding the publication of this article. reply Respond Report a concern Triet NT. Peer Review Report For: Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Techniques and Its Value Addition in Cosmeceutical Products [version 1; peer review: 1 approved, 1 not approved] . F1000Research 2025, 14 :796 ( https://doi.org/10.5256/f1000research.189257.r443296) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/14-796/v2#referee-response-443296 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2025 Khat-Udomkiri N. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 27 Sep 2025 | for Version 1 Nuntawat Khat-Udomkiri , Mae Fah Luang University, Mueang Chiang Rai, Chiang Rai, Thailand 0 Views copyright © 2025 Khat-Udomkiri N. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (1) Not Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Thank you for the opportunity to review this manuscript. The study of microwave-assisted extraction (MAE) from Plectranthus amboinicus for cosmeceutical use is topical and fits the journal scope, but the manuscript in its current form lacks novelty, methodological detail, and sufficient experimental rigor. Below I list the main weaknesses and give concrete, actionable recommendations to improve clarity, reproducibility and scientific impact. 1. You must cite and discuss prior MAE work on P. amboinicus (e.g., the paper you identified: DOI: 10.3303/CET1972067). Clearly state how your study differs (new endpoints, different solvent system, biological assays, formulation work, scale-up considerations, etc.). Right now the manuscript reads like a repeat of existing work. 2. Do not claim you “optimized” extraction conditions unless a formal design of experiments (e.g., RSM, factorial) or an explicit optimization protocol was applied. If you did not run an optimization design, rephrase to “screened”, “evaluated” or “investigated”. 3. Temperature is critical for MAE outcomes and must be reported for every condition. Report temperature profiles for all MAE runs. 4. Explain the basis for selecting caffeic acid as the principal marker. Was it the most abundant compound in the extract, the most bioactive, or selected for analytical convenience? 5. Natural extracts are mixtures — identify and quantify multiple major phenolics, not only one. Provide HPLC chromatograms and quantification for at least the top 3–5 phenolic compounds. 6. Provide a full cream formulation (all ingredients, % w/w), the exact amount of extract incorporated, and the rationale for the selected concentration(s). 7. You state extract was included during emulsification: explain and justify this choice. Phenolic compounds is heat-sensitive; if used in a heated phase, show data proving it remains active after processing. 8. You cannot assert “no degradation” based on visual inspection alone. Provide objective stability data: HPLC profiles (initial vs after processing and after stability cycles), TPC, and at least one chemical marker quantified before/after heating and after storage cycles. Replace subjective color assessments with ΔE measurements and report ΔE for color change. 9. DPPH must include a positive control (e.g., Trolox or ascorbic acid) and report results as IC₅₀ or mg Trolox equivalent/g for comparability. 10. Antimicrobial claims cannot use “broad-spectrum” without testing a representative panel. 11. Key MAE factors were omitted (type of solvent, solvent concentration, power, temperature, time, solid:liquid ratio). If you want to claim optimization, perform a DOE (factorial or RSM). Otherwise clearly present the work as a screening study. 12. Rephrase or remove any absolute claims (e.g., “broad-spectrum”, “no degradation”) unless supported by data. 13. Reword the heat-cycling sentence: heating–cooling is a preliminary shelf-stress test, not a full simulation of long-term stability. Use careful language. 14. Update and expand references: several cited works are dated — include recent MAE and cosmeceutical formulation literature. 15. Drying methods, whether heat-based (e.g., tray drying) or non-thermal (e.g., freeze-drying), are well-documented for their influence on phenolic compound stability and retention. Please clarify the rationale for selecting this factor as the focus of investigation rather than prioritizing the optimization of parameters directly affecting the extraction process. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? No Are sufficient details of methods and analysis provided to allow replication by others? No If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Partly Are the conclusions drawn adequately supported by the results? No Competing Interests No competing interests were disclosed. Reviewer Expertise Non-conventional extraction. Bioactive compounds, Cosmetics, Cell-culture assays, Formulations I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above. reply Respond to this report Responses (1) Author Response 13 Oct 2025 Doungporn Amornlerdpison, Center of Excellence in Agricultural Innovation for Graduate Entrepreneur, Maejo University, Nong Han, Thailand Reviewer 2 (Not Approved) 1. You must cite and discuss prior MAE work on P. amboinicus (e.g., the paper you identified: DOI: 10.3303/CET1972067). Clearly state how your study differs (new endpoints, different solvent system, biological assays, formulation work, scale-up considerations, etc.). Right now the manuscript reads like a repeat of existing work. Response : Our study differs by evaluating drying methods (tray vs freeze-drying), biological activities (antimicrobial and anti-inflammatory properties) and formulation into a cream with stability testing. 2. Do not claim you “optimized” extraction conditions unless a formal design of experiments (e.g., RSM, factorial) or an explicit optimization protocol was applied. If you did not run an optimization design, rephrase to “screened”, “evaluated” or “investigated”. Response : We agree and have rephrased “optimized” to “evaluated”, Page 2 unless clearly supported by data. 3. Temperature is critical for MAE outcomes and must be reported for every condition. Report temperature profiles for all MAE runs. Response: We have added the extraction temperature (40 °C under 450 W microwave power) in the Methods section, Page 3 4. Explain the basis for selecting caffeic acid as the principal marker. Was it the most abundant compound in the extract, the most bioactive, or selected for analytical convenience? Response : We selected caffeic acid as a marker because it was the most abundant phenolic identified in preliminary HPLC profiling. This justification has been added, Page 3. 5. Natural extracts are mixtures — identify and quantify multiple major phenolics, not only one. Provide HPLC chromatograms and quantification for at least the top 3–5 phenolic compounds. Response: In this study, a representative phenolic compound, caffeic acid was selected as a biomarker for quantification due to its well-documented bioactivity. Furthermore, the use of total phenolic content (TPC) provides a reliable estimation of the overall phenolic load in the extracts. This approach enables holistic evaluation of the extract's functionality while minimizing the complexity, time, and cost associated with full chromatographic profiling. Future work may include expanded HPLC profiling to better understand the full phenolic spectrum and assess potential synergistic effects among individual compounds. 6. Provide a full cream formulation (all ingredients, % w/w), the exact amount of extract incorporated, and the rationale for the selected concentration(s). Response: We have revised the section to provide the complete formulation with all ingredients and concentrations (% w/w) The formulation was based on the minimum inhibitory concentration (MIC) of PF15, determined to be 7.8 mg/mL against Staphylococcus aureus . For a 100 g cream formulation (equivalent to approximately 100 mL), the required amount of extract was calculated as 780 mg, corresponding to 0.78% (w/w) of the total formulation. The 0.8% concentration was selected to ensure antimicrobial efficacy while maintaining formulation stability, Page 5-6. 7. You state extract was included during emulsification: explain and justify this choice. Phenolic compounds is heat-sensitive; if used in a heated phase, show data proving it remains active after processing. Response: The PF15 extract was incorporated during the emulsification step to ensure uniform distribution of the bioactive compounds within the cream matrix. Although phenolic compounds are generally considered heat-sensitive, our results demonstrate that the antimicrobial activity of the extract was largely retained after processing. The PF15 extract alone showed inhibition zones of 13–15 mm (Table 3, Page 12) against selected skin pathogens, while the formulated PF cream exhibited inhibition zones of 11–12 mm (Page 16). The slight reduction indicates minor thermal impact; however, the preserved inhibitory activity confirms that the phenolic constituents remained bioactive after emulsification. 8. You cannot assert “no degradation” based on visual inspection alone. Provide objective stability data: HPLC profiles (initial vs after processing and after stability cycles), TPC, and at least one chemical marker quantified before/after heating and after storage cycles. Replace subjective color assessments with ΔE measurements and report ΔE for color change. Response: Deleted no degradation. We acknowledge the reviewer’s comment regarding the need for objective stability data. In our study, instead of quantifying individual phenolic compounds within the cream formulation, stability was evaluated based on the functional antimicrobial activity of the product. This approach was selected because the cream contains multiple ingredients. Therefore, antimicrobial efficacy against target skin pathogens was used as a practical stability indicator (Page 16), confirming that the bioactive effect of PF15 was retained after processing and throughout the stability cycles. 9. DPPH must include a positive control (e.g., Trolox or ascorbic acid) and report results as IC₅₀ or mg Trolox equivalent/g for comparability. Response : We used gallic acid as the positive control, consistent with its use in the phenolic content determination. Figure 7 has been updated to include gallic acid as the control. 10. Antimicrobial claims cannot use “broad-spectrum” without testing a representative panel. Response : Deleted broad-spectrum from conclusion. 11. Key MAE factors were omitted (type of solvent, solvent concentration, power, temperature, time, solid:liquid ratio). If you want to claim optimization, perform a DOE (factorial or RSM). Otherwise clearly present the work as a screening study. Response : We have now reported solvent type and concentration (50% ethanol), power (450 W), temperature (40 °C), extraction time (10–20 min), and solid–liquid ratio (1:10 v/v) in the Methods section, Page 3. 12. Rephrase or remove any absolute claims (e.g., “broad-spectrum”, “no degradation”) unless supported by data. Response : Statements such as “no degradation” and “broad-spectrum” have been rephrased to reflect the data accurately. 13. Reword the heat-cycling sentence: heating–cooling is a preliminary shelf-stress test, not a full simulation of long-term stability. Use careful language. Response : We have reworded this as a “preliminary stress test to evaluate its stability under accelerated conditions”, Page 16 14. Update and expand references: several cited works are dated — include recent MAE and cosmeceutical formulation literature. Response : We have updated the reference with recent MAE and other green extraction, Page 17-18 References updated: [43] 15. Drying methods, whether heat-based (e.g., tray drying) or non-thermal (e.g., freeze-drying), are well-documented for their influence on phenolic compound stability and retention. Please clarify the rationale for selecting this factor as the focus of investigation rather than prioritizing the optimization of parameters directly affecting the extraction process. Response : Our unique focus on drying and extraction methods relevance for industrial-scale processing and use and effective cost for commercial. We believe these revisions strengthen the manuscript significantly by clarifying novelty, improving methodological transparency, and ensuring accurate claims. We hope that the revised version now satisfactorily addresses all reviewer concerns and will be considered suitable for approval View more View less Competing Interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. reply Respond Report a concern Khat-Udomkiri N. Peer Review Report For: Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Techniques and Its Value Addition in Cosmeceutical Products [version 1; peer review: 1 approved, 1 not approved] . F1000Research 2025, 14 :796 ( https://doi.org/10.5256/f1000research.181624.r413222) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/14-796/v1#referee-response-413222 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2025 Srivilai J. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 15 Sep 2025 | for Version 1 Jukkarin Srivilai , University of Phayao, Phayao, Thailand 0 Views copyright © 2025 Srivilai J. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (1) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions The manuscript titled “Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Technique and Its Value Addition in Cosmeceutical Products” presents interesting and technically relevant work. However, several issues need to be addressed before indexing. The following comments are provided for the authors’ consideration: Title: The word “Techniques” should be singular (“Technique”). Abstract: In the Methods section, the last sentence should be revised to: “Anti-inflammatory potential of PF15 ….” Dosage form: The authors should clearly state the dosage form of the cream throughout the manuscript, or at least specify somewhere that it is an emulsion type. Introduction: In the third paragraph, please verify reference [6] and revise the sentence for better grammatical clarity. Materials and Methods: The description of the 50% ethanol extraction should specify the solvent used (e.g., DI water or another solvent). Chemical analysis (HPLC): The methodology should clarify whether standard compounds were used to construct the calibration curve for determining the relative amount of caffeic acid, or if another approach was applied. Please provide details. Table 1: The column “Mode” should be labeled “Mode of elution.” Abbreviations: Ensure that abbreviations follow the full name consistently (e.g., Minimal Inhibitory Concentration (MIC)). Please check throughout the manuscript. Cream formulation with PF15: Provide more details on the formulation ratio, type of emulsion, and preparation technique. Results and Discussion: Verify the unit of yield, as it may be incorrect. In Figure 3, the retention time of the caffeic acid standard does not match the retention time mentioned in the Results and Discussion section. Please correct this, as it is a critical point for analysis. Comparative discussion: Expand the discussion to include other extraction techniques or green solvents. You may cite the following articles: [Reference 1] [Reference 2] Scientific names: On page 10, in the antioxidant activity section, the correct scientific name is P. amboinicus . Similarly, check all bacterial names and ensure consistency throughout the manuscript. Table 2: Use a big letter instead of “inhibition” to be Inhibition. The caption should specify the sample/disc amount (e.g., 5 mg/disc). Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes References 1. Saesue K, Matwangsang S, Rungsang T, Khorana N, et al.: Deep eutectic-based microemulsion: A multifunctional system for enhancing extraction efficiency, stability and antityrosinase, antioxidant activities of oxyresveratrol from Artocarpus lakoocha Roxb. Journal of Molecular Liquids . 2025; 436 . Publisher Full Text 2. Rungsang T, Aimjongjun S, Aoonboontum P, Matwangsang S, et al.: A novel deep eutectic and eutectic-based microemulsion systems for enhanced extraction, stabilization and antioxidant activity of artocarpin, isocyclomorusin, and cycloartocarpin from agricultural byproduct of Artocarpus heterophyllus Lam. Microchemical Journal . 2025; 217 . Publisher Full Text Competing Interests No competing interests were disclosed. Reviewer Expertise Plant extraction technologies I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (1) Author Response 13 Oct 2025 Doungporn Amornlerdpison, Center of Excellence in Agricultural Innovation for Graduate Entrepreneur, Maejo University, Nong Han, Thailand We would like to sincerely thank the reviewers for the valuable and constructive comments. We have carefully revised the manuscript according to all suggestions, and below we provide a point-by-point response. The changes have been made as highlighted in “red font” in revised manuscript. We hope our revision has improved the paper to a satisfactory level. Reviewer 1 (Approved) 1. Title: The word “Techniques” should be singular (“Technique”). Corrected to “Technique.” 2. Abstract: In the Methods section, the last sentence should be revised to: “Anti-inflammatory potential of PF15 ….” Revised: “Anti-inflammatory potential of PF15 was evaluated in LPS-stimulated THP-1 macrophages.” 3.Dosage form: The authors should clearly state the dosage form of the cream throughout the manuscript, or at least specify somewhere that it is an emulsion type. Stated in Part: Product Development and Evaluation, Page 5 “The extract was incorporated into the cream formulation at a concentration of 0.8%, ensuring antimicrobial efficacy. The aqueous phase containing PF15 was heated to 70 °C, combined with the oil phase, and emulsified for 30 minutes. The cream was then cooled to 35–40 °C, mixed until homogeneous, and stored for further analysis. A topical cream was developed by incorporating the PF15 extract into a water-in-oil emulsion system. The detailed composition of the cream is presented in Table 2.” 4.Introduction: In the third paragraph, please verify reference [6] and revise the sentence for better grammatical clarity. Verified and corrected sentence, Page 2. “A recent study demonstrated the antimicrobial efficacy of P. amboinicus leaf extract against pathogens such as Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, and the acne-associated bacterium Cutibacterium acnes, thereby reinforcing its potential application in cosmetic formulations. 6 5.Materials and Methods: The description of the 50% ethanol extraction should specify the solvent used (e.g., DI water or another solvent). Clarified as ethanol + deionized water, Page 3. “The 50% ethanol solution was prepared using a 1:1 mixture of ethanol and deionized water.” - Chemical analysis (HPLC): The methodology should clarify whether standard compounds were used to construct the calibration curve for determining the relative amount of caffeic acid, or if another approach was applied. Please provide details. Clarified as “The standard of caffeic acid was used to construct the calibration curve.” - Table 1: The column “Mode” should be labeled “Mode of elution.” Corrected. Mode of elution - Abbreviations: Ensure that abbreviations follow the full name consistently (e.g., Minimal Inhibitory Concentration (MIC)). Please check throughout the manuscript. Checked and corrected; Minimal Inhibitory Concentration (MIC) - Cream formulation with PF15: Provide more details on the formulation ratio, type of emulsion, and preparation technique. Full formulation ratios, emulsion type, and preparation technique added, Page 5-6. 6.Results and Discussion: - Verify the unit of yield, as it may be incorrect. Corrected to % w/w dry weight - Figure 3, the retention time of the caffeic acid standard does not match the retention time mentioned in the Results and Discussion section. Please correct this, as it is a critical point for analysis. Corrected to 7.8 min retention time for caffeic acid. 7.Comparative discussion: Expand the discussion to include other extraction techniques or green solvents. You may cite the following articles: [Reference 1] [Reference 2] Expanded with ultrasound-assisted, pressurized liquid, and other green solvent extraction, Page 17-18 References updated: [43] View more View less Competing Interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. reply Respond Report a concern Srivilai J. Peer Review Report For: Bioactive Effects of Plectranthus amboinicus Extract Using Microwave Techniques and Its Value Addition in Cosmeceutical Products [version 1; peer review: 1 approved, 1 not approved] . F1000Research 2025, 14 :796 ( https://doi.org/10.5256/f1000research.181624.r407511) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/14-796/v1#referee-response-407511 Alongside their report, reviewers assign a status to the article: Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. 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