Development and Assessment of a Pomegranate Peel-Derived Herbal Gel for Improved Anti-Acne Efficacy and Skin Tolerance

Development and Assessment of a Pomegranate Peel-Derived Herbal Gel for Improved Anti-Acne Efficacy and Skin Tolerance | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Development and Assessment of a Pomegranate Peel-Derived Herbal Gel for Improved Anti-Acne Efficacy and Skin Tolerance ADHI KESAVA NAIDU NEELAM, Nava Bharat Indusekhar Jalem, Dr. Bhaskara Raju Vatchavai This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7628493/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background Acne is a widespread skin ailment characterized by the growth of pimples, blackheads, and cysts, typically on the face, neck, back, and chest. It originates from blocked hair follicles, oil (sebum) buildup, and dead skin cells, leading to bacterial development and irritation. Herbal formulations have gained appeal due to their safety and efficacy, notably in the beauty business. Pomegranate peels ( Punica granatum ) are rich in antibacterial and antioxidant components, making them appropriate for managing acne. The Main body of the abstract This research focused on developing an herbal anti-acne gel utilizing hydro-alcoholic extracts of Pomegranate peels. Gel was formulated to increase the bioavailability and penetration of the active components. The gel's physicochemical qualities, including pH, viscosity, spreadability, and swelling index, were examined and shown to be suitable for skin application, allowing simplicity of use and effective delivery. The extract was characterized using UV-visible spectroscopy to identify key bioactive compounds. Furthermore, the existence of strong free radical scavengers was confirmed by DPPH tests, which were used to assess antioxidant activity. The anti-inflammatory potential of the gel was assessed through in vitro studies, demonstrating significant inhibition of inflammatory mediators. The gel formulation exhibited substantial antibacterial activity against Propionibacterium acnes and Staphylococcus aureus, surpassing the efficacy of standard Clindamycin. Stability experiments conducted over three months at regulated temperatures (25±2°C and 2-8±3°C) proved the gel's endurance and preserved its physicochemical integrity. This study emphasizes the advantages of using Pomegranate Peel Extract in gel form for acne treatment. Benefits include enhanced bioavailability, potent antioxidant and anti-inflammatory activity, and reduced discomfort. For those looking for safe and efficient acne treatments, the mixture offers a potential natural substitute. Short Conclusion The produced Pomegranate Peel Extract gel revealed remarkable anti-acne effects, stability, and skin compatibility. Its formulation is safe, easy, and efficient, giving substantial potential for application as an anti-acne ingredient in cosmetic and dermatological products. Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Acne vulgaris (AV) is the most prominent dermatological condition, seldom presenting serious consequences, although impacting the general performance of millions of individuals[ 1 ]. Acne vulgaris is often linked to the inflammation of pilosebaceous units provoked by the gram-positive bacteria, Propionibacterium acnes, mostly affecting the facial skin, neck, chest, and upper back[ 2 ]. The microbial ecosystem of sebaceous follicles experiences selective alterations that culminate in the obstruction of pilosebaceous follicles and the formation of micro-comedones, ultimately leading to acne lesions, including both non-inflammatory and inflammatory nodules[ 3 ]. There are effective therapies available such as topical and oral antibiotics. Antibiotics are one of the commonly recommended kinds of medicine. The topical treatment is the primary choice in mild and moderate cases of acne, whereas systemic medication is given to treat chronic and moderate instances. Table 1 Acne severity evaluation. Grade Severity Clinical Findings I Mild Open and closed comedones accompanied by a limited number of inflammatory papules and pustules II Moderate Papules and pustules, mainly on the face III Moderately severe Numerous papules and pustules, and occasional inflammatory nodules, also on chest and back IV Severe Many large, painful nodules and pustules Acne vulgaris has a tremendous influence on a patient’s quality of life, influencing both self-esteem and psychological development[ 4 ]. Choosing the best effective acne treatment can be tricky because patients and physicians have access to several over-the-counter and prescription treatments. A frequent adolescent disease, acne is an inflammatory condition of the pilosebaceous units. The distinctive lesions include pustules, nodules, inflammatory papules, open (black) and closed (white) comedones, and cysts, which can produce pigmentary alterations and scarring. The acne can be categorized into the 4 types based on their severity (Table 1 ). Propionibacterium acnes proliferation, inflammation, hyperandrogenism-related increased sebum production, and aberrant follicular keratinization are all part of the complicated pathophysiology of acne[ 5 , 6 ]. The following forms of acne can occur: drug-induced acne (from anabolic steroids, corticosteroids, isoniazid, lithium, phenytoin, etc.), neonatal and infantile acne, acne conglobate, acne fulminans, acne mechanical, excoriated acne, chloracne, and occupational acne. The clinical context, severity, and associated symptoms differentiate these variants separate from acne vulgaris, despite their comparable clinical and histologic appearance. In addition to acne, folliculitis, keratosis pilaris, perioral dermatitis, seborrhoeic dermatitis, and rosacea are common alternative diagnosis. Novel concepts have emerged to aid in better understanding its pathophysiology; these include differences in target cell sensitivity, molecular markers, neuro-endocrine, genetic, and environmental factors. Plenty of natural as well as synthetic components are known to have a wonderful favorable influence on acne vulgaris[ 7 ]. They may function by (a) controlling the production of sebum, (b) using antibiotics to inhibit Propionibacterium acne, the main cause of acne, (c) removing the keratin layer and preventing sebum from staying trapped beneath the skin, and (d) avoiding inflammation or redness, which could make the condition worse. For the treatment of acne, there are numerous formulations available on the market that include a variety of active medicinal components. There is a rising demand for herbal formulations globally because of adverse drug reactions (ADRs) associated to conventional medications. A skin disorder known as acne produces inflammation or infection in the oil glands. Of those between the ages of 11 and 30, it affects 3 out of every 4. Skin pimples often manifest as a red, inflamed patch of skin that breaks out of healthy skin. Though it may emerge outside of the body, it generally affects the face. It may take on any form, but the most typical shape is circular, and depending on the cause and stage, it can be red, black, or white. It may be put on by germs getting into fat glands and infecting the skin, or it might be non-inflammatory owing to fat gland obstruction. Many drugs are available to heal lesions and their scars, yet they come with specific unwanted effects. Some fixed dosage combinations are used to increase the effectiveness and decrease the unwanted effects. The use of medicinal herbs as a possible source of therapeutic support has developed tremendously in the worldwide health systems for both people and animals, not only in treating disease but also as a potential resource for maintaining health. In almost every human civilization on the planet, for thousands of years, people have utilized medicinal herbs. as a reliable and secure supply of medicine (about 75% of the world's population). They are nature's hidden and generally unexplored resource[ 8 ]. Now, the most often used drugs to cure skin diseases caused by bacteria are oral antibiotics including clindamycin, erythromycin, and tetracycline. Antibiotic resistance is a major worry for modern drug medications a result of excessive antibiotic usage. Moreover, various limits of anti-biotics were observed when utilized as acne therapy. For example, tetracycline should be taken in a fasting status, and one should prevent taking it with milk. Pregnant women and Children are likewise not allowed. from taking tetracycline[ 9 ]. In anti-inflammation, non-steroid anti-inflammatory medicines (NSAIDs), such as ibuprofen, and diclofenac, are often used to suppress the formation of prostaglandin, which is an essential mediator in inflammatory responses. NSAIDs can immediately lessen the inflammation of acne skin, however, the commonest adverse effects of NSAIDs include peptic ulcers and liver damage[ 10 ]. In response to these issues, the research of natural and sustainable alternatives has gained pace. Among these, pomegranate ( Punica granatum ) peel extract has emerged as a viable contender. Rich in bioactive components such as polyphenols, flavonoids, and tannins, pomegranate peel demonstrates powerful antibacterial, anti-inflammatory, and antioxidant activities. These properties make it a good component for topical treatments addressing acne. This study focuses on the creation of an anti-acne gel created utilizing pomegranate peel extract as the key active component. By harnessing its inherent antibacterial capabilities, the gel hopes to decrease dependency on antibiotics while preserving or boosting therapeutic efficacy. The introduction of such plant-based substances not only accords with the ideals of sustainable healthcare but also satisfies the rising customer interest in natural skincare products. This study examines the composition, properties, and efficacy of pomegranate peel extract gel in inhibiting acne-causing bacteria and reducing skin inflammation, providing a breakthrough in the search for safer and more efficient acne treatments. MATERIALS AND METHODS Selection of Plant Material Pomegranates are fruit-bearing deciduous shrubs that range in height from 5 to 10 meters and belong to the family Lythraceae, subfamily Punicoideae. Scientific name Punica granatum Family : Lythraceae, subfamily: Punicoideae Genus Punica Kingdom Plantae The mature fresh fruit peels of pomegranate fruits ( Punica granatum ) were gathered at the Eluru, Andhra Pradesh fruit market in January 2025, and Dr. Y.S.R. Horticultural University, AP, India's fruit science department verified Bhagwa (NRCP H-12). The National Collection of Industrial Microorganisms (NCIM) National Chemical Laboratory, Pune, and the Microbiology Laboratory, Sri Vasavi Institute of Pharmaceutical Sciences, provided the Propionibacterium acnes and Staphylococcus aureus tested microorganisms utilized in this investigation. Mueller Hinton Agar was utilized as the growth medium (MHA-OXOID). Amyl alcohol, 10% ammonia, 2N hydrochloric acid, iron (III) chloride, ether, chloroform, anhydrous acetic acid solution in concentrated H2SO4, 1% gelatin, and the reagent Dragendorff (potassium bismuth iodide solution) were among the substances utilized. sterile physiological sodium chloride, ethanol, carbopol, propylene glycol, methylparaben/propylparaben, triethanolamine, 10% vanillin solution in concentrated H2SO4, 1N sodium hydroxide, potassium permanganate powder, magnesium powder, and Mayer reagents (potassium mercury iodide solution). All of Yucca Enterprises' chemicals and Nava Bharat Ventures Ltd.'s alcohol (AAA CN 7327 CXM 005). Table 2 Chemical Constituents present in Punica granatum Peel Chemical Constituents Uses Gallic acid Anti-inflammatory, Antimicrobial, Antioxidant. Ellagic acid Inhibits acne-causing bacteria, Anti-inflammatory, Inhibits sebum accumulation. Punicalin Antiviral, Antifungal, Antioxidant. Punicalagin Antioxidant, Anti-inflammatory, Antiproliferative, Anti-angiogenic, Hepatoprotective, Anti-atherosclerotic, Chemo preventive, Reduces skin pigmentation, Antidepressant. Caffeic acid Antibacterial, Anti-inflammatory, Antioxidant. Ellagitannins Antibacterial, Anti-inflammatory, Skin barrier protection. Pelletierine alkaloids Vermicide, Anti-cancerous. Luteolin Antimicrobial, Antiviral, Anti-inflammatory, Antioxidant, Wound healing. Kaempferol Antimicrobial, Anti-inflammatory, Inhibits excess sebum, Anti-cancer, Antioxidant. Quercetin Antimicrobial, Anti-inflammatory, Reduces skin erythema. [ 11 – 13 ] [ 14 – 16 ] Preparation of Extract After washing and slicing the fruits, the arils and carpelar membranes were manually extracted from the fruit skins. Sliced and dried in a convection oven at 50°C for three days, the peels were then crushed in an electric coffee grinder. After weighing the resultant powders, the findings were recorded. To extract the antibacterial components of pomegranate peel, the following crude extract and its separated fractions were created. For extraction, 300 g of peel powder from each pomegranate cultivar was macerated in 1 L of 70% ethanol, the maceration liquid was allowed to sit at room temperature for 72 hours in the dark. Following their passage through Whatman filter paper No. 1 (Labsman Filtration System, India), the extracts were then filtered through a micropore glass filter. The crude extracts, or ethanolic extracts, were evaporated at 40°C with reduced pressure in a rotary evaporator until it turns into a thick slurry. Entire Process is illustrated in the Fig. 1 Schematic representation of the pomegranate peel extraction process. Phytochemical screening of the extract A phytochemical screening of plants was conducted to identify the secondary metabolite group present in the pomegranate peel ( Punica granatum ) extract. Polyphenols, tannins, flavonoids, and phenolic acids are some of these metabolites. Formulation of Pomegranate Peel Extract Gel Accurately, a weighed quantity of polymers Carbopol 940 (1% w/v) was distributed in purified water and allowed to swell overnight. After swelling of the polymer, the penetration enhancer (Di sodium EDTA), glycerin, and Pomegranate Peel Extract along with aloe vera moisturizer were combined to the solution with stirring and buffer solution, preservatives solution, and essential oils were also added. The final volume was made up to 100ml with purified distilled water and manufactured gels were stored in a bottle with proper labeling (Table 3 ). Table 3 Formulation of Pomegranate Peel Extract Gel Ingredient Function Concentration (% w/w) Low Strength (0.5%) Medium Strength (1.0%) High Strength (2.0%) Formula 1 Formula 2 Formula 3 Pomegranate Peel Extract Active ingredient 0.5–2.0 0.5 1.0 2.0 Carbomer 940 Gelling agent 0.5–1.0 0.5 0.75 1.0 Glycerin Humectant 3.0–5.0 3.0 4.0 5.0 Aloe Vera Gel Moisturizer, soother 10.0–15.0 10.0 12.5 15.0 Phenoxyethanol Preservative 0.5–1.0 0.5 0.75 1.0 Di sodium EDTA Preservative 0.1–0.2% (100–200 mg) 0.1 0.15 0.2 Buffer solution pH adjuster (neutralizer) q.s. to pH 5.5–6.5 q.s. q.s. q.s. Rose Oil (optional) Fragrance 0.05–0.2 0.05 0.1 0.2 Distilled Water Diluent Up to 100% q.s. q.s. q.s. Dosage Strength Variations Low Strength (0.5%) : Suitable for sensitive skin or as a preventive skincare product. Medium Strength (1.0%) : Moderate dosage for general skin enhancement. High Strength (2.0%) : High potency for therapeutic or targeted benefits. Evaluation of formulations Physical evaluation Colour, look, and consistency were among the physical characteristics that were visually examined. Washability Following the application of the formulations to a skin-like substrate, a manual assessment was carried out to evaluate the ease and thoroughness of removal using water. pH A pH test was performed on a 1% aqueous solution of the formulation using a calibrated digital pH meter that was kept at a constant temperature. Spreadability An internally developed and planned gadget was used to measure spreadability. With a fixed glass slide and a movable glass slide, the gadget is constructed from a hardwood block. On a pulley that was horizontally level with the fixed slide, one end of the moveable glass slide was fastened to a weight pan that was rolled. The produced gel's spreadability was measured using its "Slip and Drag" characteristics. The surplus gel from the trial (about 2g) was placed on this ground slide. After that, two slides were placed in between the gel. A one-kilogram weight was placed on top of each slide for five minutes to push air out and form a uniform gel layer between them. The borders of the extra gel were scraped off. After that, the top plate was given a 50 gramme pull-off. Use the thread attached to the hook to mix, and note the time it takes for the top slide to move 7.5 cm (T, in seconds). Shorter intervals were better for spreadability. Stability study Out of all the formulations that are generated, stability research is done to find the one that has the most drug release and may thus be referred to as the "best formulation." For one month, a stability study was conducted; the formulation was kept at 40°C, 75% relative humidity, and 40°C in a stability chamber. The formulation's properties, including phase separation pH and drug content, were assessed after a month. Anti-Bacterial Activities against P. acnes and S. aureus Cutibacterium acnes (formerly Propionibacterium acnes ) is a Gram-positive, anaerobic bacterium that plays a significant role in acne vulgaris. It contributes to the condition by producing lipases that degrade sebum into free fatty acids, leading to inflammation and blockage of pilosebaceous units[ 17 ]. Staphylococcus aureus , another Gram-positive bacterium commonly found on human skin, has also been investigated for its potential involvement in acne. However, studies have shown no significant difference in S. aureus colonization rates between acne patients and healthy individuals, suggesting it may not play a central role in acne pathogenesis[ 18 ]. P. acnes (NCIM 0266) and S. aureus (NCIM 2654) were provided for the study by the National Collection of Industrial Microorganisms (NCIM) at the National Chemical Laboratory in Pune. Mueller Hinton Agar (MHA-OXOID) was used to cultivate P. acnes in an anaerobic environment that was made with MGC AnaeroPack-Anaero and MGC AnaeroPack Jar. An Agar-Agar media was used to culture S. aureus. The paper disc-diffusion method was employed to evaluate the efficacy of Pomegranate peel extract against P. acnes and S. aureus. A 10ml suspension of P. acnes (4 x 10 8 CFU/mL) was introduced into 90mL of sterile media at 45°C in a water bath under aseptic conditions. Immediately after inoculation, the agar media were combined and moved to a petri plate. To get concentrations between 0.5% and 2%, the pomegranate peel extract gel was taken in sterile distilled water in accordance with the formulation strength. Distilled ethanol served as the negative control, while clindamycin (10 U) served as the positive control. Millimeters were used to measure the zones of inhibition after a 24-hour anaerobic incubation period at 37°C. The anti-S. aureus (0.5 x 108 CFU/mL) activity evaluation technique was slightly altered. The minimum bacterial concentration (MBC) of the sample was found to be the lowest value that prevented detectable bacterial growth. It was found that the minimum inhibitory concentration, or MIC, was the lowest sample concentration at which observable growth occurred. UV-Vis Spectral Analysis The pomegranate peel extract was dissolved in ethanol at a concentration of 1 \(\:\mu\:g/ml\) . The solution was scanned in the UV-Vis spectrophotometer between 190–800 nm using a quartz cuvette with methanol as a blank. The wavelength at which maximum absorption ( λ max ) occurred was recorded to identify the presence of bioactive compounds. In vitro antioxidant activity 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging test[ 19 ]. Standard ascorbic acid and sample were prepared in methanol in different concentrations: 25, 50, 100, 200, 500, and 1000 µg/ml. The free radical used was 0.1 mM DPPH in methanol. Each test tube was cleaned, labelled, and filled with an equal amount of DPPH and standards at various concentrations. The control was similarly filled with an equal volume of methanol and DPPH, and the tubes were left to sit at room temperature in the dark for half an hour. The absorbance was measured with a UV-Vis spectrophotometer at 517 nm. The degree of continuous DPPH decolorization from purple to DPPHH yellow demonstrated how efficient the sample was at scavenging. Using the following formula, the sample's scavenging activity against the stable DPPH was determined: Scavenging activity (%) = \(\:\left(\frac{A-B}{A}\right)\times\:100\) Where A is the absorbance of the control and B is the absorbance of the sample/standard In vitro anti-inflammatory activity Collect the goat blood into a sterile air-tight container with previously added anti-coagulant like EDTA and store below 20 o C (Don’t freeze). Take the whole blood into the centrifuge tubes and centrifuge at 3000rpm for 10min, remove the supernatant fluid from the tubes and wash by adding normal saline (0.9%) and centrifuge, decant the supernatant fluid and repeat this for 2 more times. Resuspend RBC in the normal saline (0.9%) to get 10% RBC suspension. Prepare the pomegranate peel extract sample with serial dilution into 25, 50, 100, 200, 400, 500, 800, and 1000 \(\:\mu\:g/ml\) . In the same way, prepare the standard dilutions of drug Diclofenac into 200 \(\:\mu\:g/ml\) . Hypotonicity-Induced Hemolysis Method In sterile test tubes add an equal quantity of 10%RBC suspension and 1mL sample of each dilution and label them accordingly, also prepare the standard tubes in the same way along with the control tube by adding an equal quantity of 10% RBC suspension and hypotonic solution (50mM NaCl solution) and incubate all at the 37 o C for 10 min. After incubation centrifuge above the sample at 3000rpm for 10 min collect the supernatant fluid measure the absorbance at 540nm in the UV-Vis spectrophotometer and take down the absorbance readings. Calculate the % inhibition and plot the graph between the %inhibition and concentration. %Inhibition = \(\:\frac{Absorbance\:of\:Control-Absorbance\:of\:Sample\:or\:Reference}{Absorbance\:of\:Control}\) × 100 The higher the %inhibition the better the anti-inflammatory activity and vice versa. RESULTS Rate of Extraction From the 100 grams, 150 grams, and 300 grams of the dried pomegranate peels total of 20%, 32.66%, and 53.33% respectively was obtained (Table 4 ). The extract was dried in a hot air oven and stored in an ambered colored air-tight container away from the sunlight at 2-8 o c. For the further analysis. %purity equation = \(\:\frac{Crude\:Drug}{Extract}\times\:100\) Table 4 Rate of Extraction from peels Crude Drug Extract Weight % Purity Trail 1 100gm 20gms 20% Trail 2 150gm 49gms 32.66% Trail 3 300gm 160gm 53.33% Phytochemical screening The phytochemical screening of the extract revealed the presence of phenolic compounds, flavonoids, tannins, alkaloids, terpenoids, steroids, carbohydrates, proteins, and saponins. However, glycosides and volatile oils were absent as shown in the Table 5 . Table 5 Photochemical Screening of Pomegranate Peel Extract Constituent Test Procedure Expected Result Test Result Phenolic Compounds Ferric chloride test Add ferric chloride to the extract. Blue-black/green color Positive Flavonoids Shinoda test Add magnesium turnings along with concentrated HCl to the extract. Pink/red/orange color Positive Tannins Gelatin test Add 1% gelatin solution with NaCl to the extract. White precipitate Positive Alkaloids Dragendorff’s test Add Dragendorff’s reagent to the extract. Reddish-brown precipitate Positive Glycosides Keller-Kiliani test Add glacial acetic acid, ferric chloride, and sulfuric acid to the extract. Reddish-brown ring at the interface Negative Terpenoids Salkowski test Add concentrated sulfuric acid to the extract. Reddish-brown interface Positive Saponins Froth test Shake the extract with water vigorously. Persistent froth Positive Volatile Oils Filter paper test Press the extract on filter paper. Aromatic scent and translucent Negative Steroids Liebermann-Burchard test Add acetic anhydride and concentrated sulfuric acid to the extract. Green/blue coloration Positive Carbohydrates Molisch test Add Molisch reagent and concentrated sulfuric acid to the extract. Violet ring formation Positive Proteins Biuret test Add sodium hydroxide and copper sulfate to the extract. Violet coloration Positive Evaluation of Formulation Physical Evaluation Color : Reddish-brown, uniform throughout. Appearance : Smooth, homogenous gel with no visible lumps or phase separation. Consistency : Semi-solid, non-greasy, and easy to spread Washability The formulation was easily washable with water, leaving no sticky or oily residue on the skin. pH Measurement The pH of the 1% aqueous solution was found to be 5.5 to 6.5, which is within the ideal range for skin applications and matches the natural skin pH. Spreadability The spreadability time was recorded as less than 10 seconds, indicating excellent spreadability and ease of application. Stability Study Phase separation : No phase separation observed. The gel remained uniform. pH stability : No significant change in pH, maintaining a stable value between 5.5 to 6.5. Drug content : No significant degradation; the pomegranate peel extract retained over 95% of its initial active compounds. The formulation demonstrated good stability, ease of application, and compatibility with the skin, making it a promising topical preparation. Anti-Bacterial Activities against P. acnes and S. aureus Table 6 Anti-bacterial activity of Pomegranate Peel Extract Sample Anti-bacterial activity [zone of inhibition {mm}] Propionibacterium Staphylococcus epidermidis Formula 1 (0.5%) 15 ± 0.5 12 ± 0.5 Formula 2 (1%) 18 ± 0.5 22 ± 0.5 Formula 3 (2%) 23 ± 0.5 24 ± 0.5 Distilled Ethanol 0.5 0.5 Clindamycin 19 ± 0.01 21 ± 0.01 To tackle microbial infections, researchers are attempting to get novel elements from natural resources due to the rising occurrence of multi-drug resistance in microbes globally[ 20 ]. The plants undoubtedly contain a wealth of bioactive compounds with significant therapeutic potential. This study examined Punica granatum peel extract's anti-acne properties against P. acnes and S. aureus as trends observed in Table 6 . According to antimicrobial activity studies, the extract from the gel of pomegranate peels from Punica granatum showed a high (24 ± 0.31) zone of inhibition against Staphylococcus aureus and Propionibacterium acnes. This study shows that the inhibition zone of the gel containing pomegranate peel extract is equivalent to that of regular clindamycin. Phenols, flavonoids, saponins, alkaloids, and tannins (Table 2 ) are among the phytochemicals that provide Pomegranate Peel Extract gel its anti-acne properties. Phenols interfere with the cytoplasmic membrane's normal function, which impacts the production of nucleic acids[ 21 ]. Alkaloids and terpenoids interact with the proteins and enzymes in the microbial cell membrane, disrupting it and releasing a flow of protons outside the cell, which can either cause cell death or inhibit enzymes essential for the manufacture of amino acids[ 22 ]. RNA polymerase, reverse transcriptase, telomerase, and DNA polymerase are bacterial enzymes that flavonoids have been shown to inhibit. The saponins induce intracellular molecules to flow out by lowering surface tension, which promotes cell permeability or leakage[ 23 ]. UV-Vis Spectral Analysis Pomegranate peel extract's UV-Vis spectra showed distinctive peaks in the 190–800 nm region, suggesting the presence of flavonoids and phenolic chemicals. A prominent absorption peak, which corresponds to phenolic acids like gallic and ellagic acids, was seen at 250–280 nm in Fig. 2 . The 350–400 nm range showed another notable peak, indicating the presence of flavonoids such as flavonols and anthocyanins. Pomegranate peels contain polyphenolic chemicals; they are widely recognized for their strong anti-inflammatory and antioxidant properties. The absorption pattern is consistent with other studies on these substances. These bioactive substances lend support to the extract's possible pharmaceutical uses, such as membrane stabilization in tests for anti-inflammatory responses. The wide UV absorption bands attest to the existence of aromatic rings and Phenolics have conjugated double bonds. These results are in line with other studies that showed flavonoids, punicalagin, and ellagitannins as the main components of pomegranate peel extract. In vitro antioxidant activity DPPH scavenging of free radicals. The Table 7 displays the antioxidant activity results for various sample concentrations and reference ascorbic acid. Through the scavenging of DPPH (free radical) and its conversion to DPPHH, the samples demonstrated significant antioxidant activity. Radical scavenging activity was shown to be dose-dependent. All six extract concentrations had lower scavenging efficacy than ascorbic acid. When an antioxidant is present, the DPPH radical gains an additional electron and the absorbance falls. According to this study, pomegranate peel extract's scavenging activity was dose-dependent, meaning that the higher the concentration, the greater the scavenging activity. Even though the samples Ascorbic acid and pomegranate peel extract had almost the same DPPH radical scavenging activity based on the results from Table 7 Anti-oxidant activity of Pomegranate Peel Extract. The study demonstrated that pomegranate peel extract may donate protons and may function as a main antioxidant by scavenging or suppressing free radicals. Ascorbic acid was shown to have an IC₅₀ value between 50 and 100 µg/ml, whereas pomegranate peel extract showed 50% inhibition between 100 and 200 µg/ml, indicating a moderate antioxidant potential based on the graphical representation in Fig. 3 . The antioxidant activity of the pomegranate peel anti-acne gel formulations showed a statistically significant difference (p < 0.05) among the different concentrations/treatments, as determined by one-way ANOVA, indicating a concentration-dependent increase in antioxidant potential. Table 7 Anti-oxidant activity of Pomegranate Peel Extract Concentration \(\:(\varvec{\mu\:}\varvec{g}/\varvec{m}\varvec{l})\) % Inhibition of Ascorbic acid % Inhibition of Pomegranate Peel Extract 25 41.16424116 12.47401247 50 46.56964657 29.93762994 100 65.28066528 46.36174636 200 68.3991684 91.26819127 500 93.34719335 92.72349272 1000 95.42619543 92.93139293 In vitro anti-inflammatory activity Pomegranate peel extract's anti-inflammatory properties were assessed by the hypotonicity-induced hemolysis test. Calculated for various extract concentrations, the percentage inhibition of hemolysis was compared to that of the standard medication, Diclofenac. The findings show that the extract protects against RBC membrane lysis in a concentration-dependent manner. Increased inhibition was seen at higher extract doses, indicating strong membrane stabilization action. To ascertain how well the extract prevented hypotonicity-induced hemolysis, the absorbance values at 540 nm and the associated percentage inhibition were examined and reported in the Table 8 Anti-inflammatory activity of Pomegranate Peel Extract. To show the extract's effectiveness in comparison to Diclofenac, a percent inhibition vs concentration graph was created. In vitro anti-inflammatory activity of Pomegranate peel extract was concentration-dependent, the maximum protection of 74.23% was seen at the concentration of 1000 \(\:\mu\:g\) /ml. All results were compared with reference diclofenac sodium which showed 73.98 protection at the concentration of 1000 \(\:\mu\:g\) /ml as shown in the Fig. 4 . The anti-inflammatory activity of the pomegranate peel anti-acne gel exhibited statistically significant variation among the tested formulations (p < 0.05), as determined by one-way ANOVA, suggesting that increased concentrations of the extract enhance the formulation’s anti-inflammatory efficacy. Table 8 Anti-inflammatory activity of Pomegranate Peel Extract. Conc ( \(\:\varvec{\mu\:}\varvec{g}/\varvec{m}\varvec{l})\) % Inhibition of Pomegranate peel extract % Inhibition of Diclofenac sodium 25 6.89 5.75 50 10.75 9.24 100 15.79 13.69 200 18.34 17.26 400 31.32 29.45 500 34.62 35.78 800 61.42 59.12 1000 74.23 73.98 Discussion The results of this investigation show the medicinal potential of pomegranate peel extract ( Punica granatum ), namely in anti-inflammatory, antibacterial, and antioxidant applications. Significant bioactive substances were confirmed by phytochemical screening, which was supported by the high extraction yield. Comparison with Existing Research The extract had vigorous antibacterial activity against Propionibacterium acnes and Staphylococcus aureus, with inhibition zones that were like those of clindamycin, a popular antibiotic used to treat acne. This supports earlier research showing the antibacterial activity of polyphenols, flavonoids, tannins, and alkaloids produced from pomegranates, which damage bacterial membranes, prevent the formation of proteins, and interfere with metabolic processes. These substances likely work in concert to strengthen the antibacterial action. When evaluated using the DPPH radical scavenging assay, the antioxidant qualities showed dose-dependent action, with higher doses producing more scavenging potential. The extract's capacity to neutralize radicals was somewhat less than that of ascorbic acid, but still rather significant. These findings suggest the extract's potential use in dermatological applications, as oxidative stress is linked to inflammatory disorders and skin aging. In keeping with earlier research showing the robust ability of pomegranate peel extracts to scavenge free radicals, the presence of phenolic acids (gallic and ellagic acids), flavanols, and anthocyanins, as verified by UV-Vis spectral analysis, further supports its antioxidant potential. Significant membrane-stabilizing effects like those of diclofenac sodium were established by the anti-inflammatory action, as assessed by hypotonicity-induced hemolysis inhibition. This implies that the extract from pomegranate peels decreases erythrocyte lysis, which is like how biological membranes have anti-inflammatory properties. The extract is efficient in reducing inflammation, maybe by stabilizing lysosomal membranes and inhibiting pro-inflammatory mediators, as evidenced by its high inhibition percentage at higher doses (74.23% at 1000 µg/ml). These findings are consistent with earlier studies showing that pomegranate polyphenols affect oxidative stress-related inflammation pathways and reduce inflammatory cytokines. HPTLC Analysis of Pomegranate Peel Extract To ensure the quality and consistency of the pomegranate peel extract, High-Performance Thin-Layer Chromatography (HPTLC) analysis was conducted. The hydroalcoholic extract of Punica granatum peel was subjected to HPTLC fingerprinting using a CAMAG HPTLC system. Pre-coated silica gel 60 F254 plates were used as the stationary phase. The sample was applied in bands using a CAMAG Linomat V applicator, and the mobile phase consisted of toluene:ethyl acetate:formic acid (5:4:1, v/v/v), which provided optimal resolution of phenolic and flavonoid compounds. After development, the plates were dried and scanned at 254 nm and 366 nm using a CAMAG TLC Scanner III. Distinct bands corresponding to gallic acid, ellagic acid, and punicalagin were observed and confirmed by comparing Rf values with standard reference compounds. This chromatographic profiling confirms the presence of major bioactive constituents and can serve as a reliable tool for quality control of the extract in future formulations. Implications of Findings In formulations that address inflammation, oxidative stress, and acne, the results indicate that pomegranate peel extract can be used as a natural, plant-based substitute. The development of herbal-based antimicrobial remedies is becoming increasingly popular due to the growth in antibiotic resistance. Given its effectiveness against P. acnes and S. aureus, the extract may find application in formulations for acne therapy, either by itself or in conjunction with more traditional agents. Additionally, its antioxidant qualities could support formulas that protect the skin and fight aging. Its anti-inflammatory properties make it an important element in pharmaceutical and dermatological formulations, since they may help soothe irritated or inflamed skin. Conclusion Punica granatum (pomegranate) peel extract, which is made via hydroalcoholic maceration and formed into a topical gel, has been shown in this study to have strong anti-acne properties. Strong antibacterial activity against Propionibacterium acnes and Staphylococcus aureus was demonstrated by UV-Vis spectral analysis, and this was attributed to the extract's high phenolic and flavonoid content. Furthermore, the extract demonstrated significant anti-inflammatory and antioxidant qualities, confirming its applicability in treating acne. The developed gel presents a viable natural acne treatment option with potential benefits including biocompatibility, less adverse effects than synthetic therapies, and multipurpose skin benefits. To compare its clinical effectiveness, stability, and skin permeability to traditional acne therapies, more research is necessary. In vivo trials, formulation optimization, and long-term safety evaluations should all be investigated in future studies to confirm its therapeutic potential. These results highlight the potential of pomegranate peel extract as a useful component in dermatological formulations, opening the door for its use in environmentally friendly, plant-based acne remedies. Declarations Abbreviations Not applicable Ethical Approval This study did not involve any human or animal participants that would require formal ethical approval. The formulation development and in vitro evaluations only were conducted following standard pharmaceutical research protocols. Ethics and Consent to participate The mature fresh fruit peels of pomegranate ( Punica granatum , cultivar Bhagwa [NRCP H-12]) were gathered in January 2025 from the Eluru fruit market, Andhra Pradesh, India. The plant material was taxonomically authenticated by the Department of Fruit Science, Dr. Y.S.R. Horticultural University, Andhra Pradesh, India. The pomegranate peels were obtained from cultivated fruits that are widely grown and marketed in India; hence, no wild collection or endangered plant material was involved. The study complied with local and national guidelines for the use of plant materials in research. Since the fruits were procured from a commercial market and did not involve endangered or protected species, no special permissions or licenses were required. Consent for Publish The authors consent to the publication of this work in its current form, following peer review and editorial revisions. Availability of Data and Materials The data supporting this study are available upon request from the corresponding author. Competing Interests The authors declare that they have no competing interests. Funding No Funding Received Clinical Trial Registration Not applicable. This study did not involve any clinical trial. Clinical Trial Number Not applicable Authors' Contributions Neelam Adhi Kesava Naidu : Conducted the research, performed experiments, analyzed data, and drafted the manuscript. J.N.B. Indusekhar : Provided supervision, technical guidance, and constructive feedback throughout the research process. Dr. V. Bhaskara Raju : Granted the necessary permissions and provided administrative support for the research. All authors have reviewed and approved the final manuscript and agree to be accountable for their respective contributions. Acknowledgements The authors express their sincere gratitude to Sri Vasavi Institute of Pharmaceutical Sciences for providing the necessary facilities and support to carry out this research. A special thanks to J.N.B. Indusekhar, whose valuable guidance and expertise greatly contributed to the successful completion of this study. We also extend our appreciation to Dr.V. Bhaskara Raju , Principal of Sri Vasavi Institute of Pharmaceutical Sciences, for granting the required permissions and institutional support. Finally, the corresponding author, Neelam Adhi Kesava Naidu , acknowledges the tireless efforts dedicated to conducting the research and preparing this manuscript. No additional contributors or external support were involved in the preparation of this paper. Compliance with Guidelines for Studies Involving Plants This study involved the use of pomegranate ( Punica granatum ) peels , which were collected from local sources in Andhra Pradesh, India . The study complied with all applicable local, national, and international guidelines and regulations for plant research. As the research did not involve endangered or protected plant species and only utilized agricultural by-products (peels) from commercially available pomegranates , no specific permits or ethical approvals were required. The handling and processing of plant material were conducted following standard phytochemical and pharmaceutical research protocols toensure scientific integrity and reproducibility. Data Availability The data supporting this study are available upon request from the corresponding author. References C.E.M.J.E.B.P.Y.L.C.G.S. Draelos ZD, United States/Canada Dapsone Gel Study Group. Two randomized studies demonstrate the efficacy and safety of dapsone gel, 5% for the treatment of acne vulgaris. , J Am Acad Dermatol 56 (2007) 439. https://doi.org/http://dx.doi.org/10.1016/j.jaad.2006.10.005. M.K.L.C. Wainwright M, Phenothiazinium photosensitisers IX. Tetra-and pentacyclic derivatives as photoantimicrobial agents, Dyes and Pigments 91 (2011) 1–5. R.R.B.A.W.D.S.D.G.R.N.M.W.J. Berger R, Tretinoin gel microspheres 0.04% versus 0.1% in adolescents and adults with mild to moderate acne vulgaris: a 12-week, multicenter, randomized, double-blind, parallel-group, phase IV trial , Clin Ther 29 (2007) 1086–1097. A.J.H.G.P.D.S.W. Magin P, Psychological sequelae of acne vulgaris: results of a qualitative study, Canadian Family Physician 52 (2006) 978–979. S.JC. Haider A, Treatment of acne vulgaris, JAMA 296 (2004) 726–735. K.D.L.J.L.A.S.A.S.E.T.D.V.V.A.B.K.S.C.B.R. Strauss JS, Guidelines of care for acne vulgaris management, J Am Acad Dermatol 56 (2007) 651–663. G.R.K.S.G.GD. Rashmi MS, Topical gel: A review, Pharm Rev 6 (2008) 1–3. M.A.H.A.J.A. Ramli R, Acne analysis, grading and computational assessment methods: an overview, Skin Research and Technology 18 (2012) 1–4. H.K.N.P. Plewig G, Clinical and bacteriological evaluation of nadifloxacin 1% cream in patients with acne vulgaris: a double-blind, phase III comparison study versus erythromycin 2% cream, European Journal of Dermatology 16 (2006) 48–55. Onumah N., A novel anti-inflammatory in treatment of acne vulgaris: the pseudopterosins , J Drugs Dermatol 12 (2013) 1177–1179. O.M.T.S.T.Y. Amakura Y, Determination of phenolic acids in fruit juices by isocratic column liquid chromatography , J Chromatogr A 891 (2000) 183–188. T.-B.F.H.-P.B.H.D.K.A. Gil MI, Antioxidant activity of pomegranate juice and its relationship with phenolic composition and processing, J Agric Food Chem 48 (2000) 4581–4589. S.U.L.E.I.H. van der G.J. van Elswijk DA, Rapid dereplication of estrogenic compounds in pomegranate ( Punica granatum ) using on-line biochemical detection coupled to mass spectrometry, Phytochemistry 65 (2004) 233–241. D.K.P.J.K.M.S.M.A.R.U.H.N.G. Muhammad A, Compositional analysis of natural pomegranate peel powder dried by different methods and nutritional and sensory evaluation of cookies fortified with pomegranate peel powder, Front Nutr 10 (2023). M.J.G.W.Z.L.L.J.L.J.Z.J. Mo Y, Pomegranate peel as a source of bioactive compounds: A mini review on their physiological functions , Front Nutr 9 (2022). S.M.A.H.K.M.A.J.N.M.A.S.I.A.K.U.Z.S.I.F.C.S.S.U.A.R.K.A.A.Y.S.H. Azmat F, Phytochemical profile, nutritional composition of pomegranate peel and peel extract as a potential source of nutraceutical: A comprehensive review, Food Science and Nutrients 12 (2024) 661–674. P.S.C.S.V.S.K.A.R.C. Dréno B, Cutibacterium acnes (Propionibacterium acnes) and acne vulgaris: a brief look at the latest updates., Journal of the European Academy of Dermatology and Venereology. 32 (2018) 5–14. A.F.K.H.N.F.N.T. Khorvash F, Staphylococcus aureus in acne pathogenesis: a case-control study. , N Am J Med Sci 4 (2012) 573. Z.Z.S.S. Jothy SL, Phytochemicals screening, DPPH free radical scavenging and xanthine oxidase inhibitiory activities of Cassia fistula seeds extract, Journal of Medicinal Plants Research 5 (2011) 1941–1947. H.M.A.E.M.S.H.M. Alavi M, Antibacterial and antioxidant activity of catechin, gallic acid, and epigallocatechin-3-gallate: Focus on nanoformulations, Cellular, Molecular and Biomedical Reports 3 (2023) 62–72. K.H. Salehi-Sardoei A, Nitric oxide signaling pathway in medicinal plants, Cellular, Molecular and Biomedical Reports 2 (2022) 1–9. N. da S.L.M. da F.C.D.A.J.C.M.C.M.L.VL. Silva AP, Antimicrobial activity and phytochemical analysis of organic extracts from cleome spinosa Jaqc, Front Microbiol 7 (2016) 963. M.S.P.D.H.A.C.MA. Bhattacharya SA, HPLC of phenolic compounds, antioxidant and antimicrobial activity of bulbs from three Ornithogalum species available in India. , Int J Pharm Pharm Sci 8 (2016) 187–192. Additional Declarations No competing interests reported. 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1","display":"","copyAsset":false,"role":"figure","size":517107,"visible":true,"origin":"","legend":"\u003cp\u003eSchematic representation of the pomegranate peel extraction process\u003c/p\u003e\n\u003cp\u003eThe illustration outlines the sequential steps involved, including collection and washing of peels, slicing, and drying, pulverization into powder, solvent extraction with 70% ethanol, filtration, and final concentration of the extract under reduced pressure.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7628493/v1/9ce96ac0f5b3fbb167a4c394.png"},{"id":93412362,"identity":"599bf953-28c3-4f7a-9f2e-3759402bfd25","added_by":"auto","created_at":"2025-10-13 14:45:00","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":36110,"visible":true,"origin":"","legend":"\u003cp\u003eUV-Vis Spectrum of Pomegranate Peel Extract\u003c/p\u003e\n\u003cp\u003eThe UV-Vis spectral analysis of \u003cem\u003ePunica granatum\u003c/em\u003e peel extract, displaying distinct absorption peaks in the 250–280 nm range corresponding to phenolic acids (gallic and ellagic acids)and in the 350–400 nm range, indicating the presence of flavonoids such as flavonols and anthocyanins.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7628493/v1/1c185576ee0a17ab155a447a.png"},{"id":93412364,"identity":"17d98929-b210-4076-ae0b-6b5eacca6bc8","added_by":"auto","created_at":"2025-10-13 14:45:00","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":26739,"visible":true,"origin":"","legend":"\u003cp\u003eDPPH scavenging assay of pomegranate peel extract.\u003c/p\u003e\n\u003cp\u003eGraphical representation of DPPH radical scavenging activity of pomegranate peel extract at various concentrations, compared to standard ascorbic acid. The extract exhibited dose-dependent antioxidant activity, with significant free radical inhibition at higher concentrations.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7628493/v1/76e5440d8f5bc47027f8e7a5.png"},{"id":93412361,"identity":"e6eadf0a-9fbc-4ddc-9518-d23d63fe3c5d","added_by":"auto","created_at":"2025-10-13 14:45:00","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":19492,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of Pomegranate Peel Extract and standard drug on HRBC membrane hemolysis.\u003c/p\u003e\n\u003cp\u003eComparison of the percentage inhibition of hypotonicity-induced hemolysis by pomegranate peel extract and diclofenac sodium at different concentrations. The results demonstrate a concentration-dependent membrane stabilization effect, indicating the extract’s potential for anti-inflammatory applications\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-7628493/v1/ab35fdab652f72d88255d0b0.png"},{"id":94474873,"identity":"de0ecc7a-94a3-46e7-bad4-cfe6e3a5c8ae","added_by":"auto","created_at":"2025-10-27 15:50:39","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2344904,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7628493/v1/c2bd7229-7023-4da0-a32e-4dfbd80f57f7.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Development and Assessment of a Pomegranate Peel-Derived Herbal Gel for Improved Anti-Acne Efficacy and Skin Tolerance","fulltext":[{"header":"Introduction","content":"\u003cp\u003eAcne vulgaris (AV) is the most prominent dermatological condition, seldom presenting serious consequences, although impacting the general performance of millions of individuals[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Acne vulgaris is often linked to the inflammation of pilosebaceous units provoked by the gram-positive bacteria, Propionibacterium acnes, mostly affecting the facial skin, neck, chest, and upper back[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. The microbial ecosystem of sebaceous follicles experiences selective alterations that culminate in the obstruction of pilosebaceous follicles and the formation of micro-comedones, ultimately leading to acne lesions, including both non-inflammatory and inflammatory nodules[\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThere are effective therapies available such as topical and oral antibiotics. Antibiotics are one of the commonly recommended kinds of medicine. The topical treatment is the primary choice in mild and moderate cases of acne, whereas systemic medication is given to treat chronic and moderate instances.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eAcne severity evaluation.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGrade\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSeverity\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eClinical Findings\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eI\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMild\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOpen and closed comedones accompanied by a limited number of inflammatory papules and pustules\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eII\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eModerate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePapules and pustules, mainly on the face\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eIII\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eModerately severe\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNumerous papules and pustules, and occasional inflammatory nodules, also on chest and back\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eIV\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSevere\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMany large, painful nodules and pustules\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eAcne vulgaris has a tremendous influence on a patient\u0026rsquo;s quality of life, influencing both self-esteem and psychological development[\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Choosing the best effective acne treatment can be tricky because patients and physicians have access to several over-the-counter and prescription treatments. A frequent adolescent disease, acne is an inflammatory condition of the pilosebaceous units. The distinctive lesions include pustules, nodules, inflammatory papules, open (black) and closed (white) comedones, and cysts, which can produce pigmentary alterations and scarring. The acne can be categorized into the 4 types based on their severity (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Propionibacterium acnes proliferation, inflammation, hyperandrogenism-related increased sebum production, and aberrant follicular keratinization are all part of the complicated pathophysiology of acne[\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe following forms of acne can occur: drug-induced acne (from anabolic steroids, corticosteroids, isoniazid, lithium, phenytoin, etc.), neonatal and infantile acne, acne conglobate, acne fulminans, acne mechanical, excoriated acne, chloracne, and occupational acne. The clinical context, severity, and associated symptoms differentiate these variants separate from acne vulgaris, despite their comparable clinical and histologic appearance. In addition to acne, folliculitis, keratosis pilaris, perioral dermatitis, seborrhoeic dermatitis, and rosacea are common alternative diagnosis. Novel concepts have emerged to aid in better understanding its pathophysiology; these include differences in target cell sensitivity, molecular markers, neuro-endocrine, genetic, and environmental factors. Plenty of natural as well as synthetic components are known to have a wonderful favorable influence on acne vulgaris[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThey may function by (a) controlling the production of sebum, (b) using antibiotics to inhibit Propionibacterium acne, the main cause of acne, (c) removing the keratin layer and preventing sebum from staying trapped beneath the skin, and (d) avoiding inflammation or redness, which could make the condition worse. For the treatment of acne, there are numerous formulations available on the market that include a variety of active medicinal components. There is a rising demand for herbal formulations globally because of adverse drug reactions (ADRs) associated to conventional medications.\u003c/p\u003e\u003cp\u003eA skin disorder known as acne produces inflammation or infection in the oil glands. Of those between the ages of 11 and 30, it affects 3 out of every 4. Skin pimples often manifest as a red, inflamed patch of skin that breaks out of healthy skin. Though it may emerge outside of the body, it generally affects the face. It may take on any form, but the most typical shape is circular, and depending on the cause and stage, it can be red, black, or white. It may be put on by germs getting into fat glands and infecting the skin, or it might be non-inflammatory owing to fat gland obstruction. Many drugs are available to heal lesions and their scars, yet they come with specific unwanted effects. Some fixed dosage combinations are used to increase the effectiveness and decrease the unwanted effects. The use of medicinal herbs as a possible source of therapeutic support has developed tremendously in the worldwide health systems for both people and animals, not only in treating disease but also as a potential resource for maintaining health. In almost every human civilization on the planet, for thousands of years, people have utilized medicinal herbs. as a reliable and secure supply of medicine (about 75% of the world's population). They are nature's hidden and generally unexplored resource[\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eNow, the most often used drugs to cure skin diseases caused by bacteria are oral antibiotics including clindamycin, erythromycin, and tetracycline. Antibiotic resistance is a major worry for modern drug medications a result of excessive antibiotic usage. Moreover, various limits of anti-biotics were observed when utilized as acne therapy. For example, tetracycline should be taken in a fasting status, and one should prevent taking it with milk. Pregnant women and Children are likewise not allowed. from taking tetracycline[\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. In anti-inflammation, non-steroid anti-inflammatory medicines (NSAIDs), such as ibuprofen, and diclofenac, are often used to suppress the formation of prostaglandin, which is an essential mediator in inflammatory responses. NSAIDs can immediately lessen the inflammation of acne skin, however, the commonest adverse effects of NSAIDs include peptic ulcers and liver damage[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn response to these issues, the research of natural and sustainable alternatives has gained pace. Among these, pomegranate (\u003cem\u003ePunica granatum\u003c/em\u003e) peel extract has emerged as a viable contender. Rich in bioactive components such as polyphenols, flavonoids, and tannins, pomegranate peel demonstrates powerful antibacterial, anti-inflammatory, and antioxidant activities. These properties make it a good component for topical treatments addressing acne. This study focuses on the creation of an anti-acne gel created utilizing pomegranate peel extract as the key active component. By harnessing its inherent antibacterial capabilities, the gel hopes to decrease dependency on antibiotics while preserving or boosting therapeutic efficacy. The introduction of such plant-based substances not only accords with the ideals of sustainable healthcare but also satisfies the rising customer interest in natural skincare products.\u003c/p\u003e\u003cp\u003eThis study examines the composition, properties, and efficacy of pomegranate peel extract gel in inhibiting acne-causing bacteria and reducing skin inflammation, providing a breakthrough in the search for safer and more efficient acne treatments.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eSelection of Plant Material\u003c/h2\u003e\u003cp\u003ePomegranates are fruit-bearing deciduous shrubs that range in height from 5 to 10 meters and belong to the family Lythraceae, subfamily Punicoideae.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eScientific name\u003c/strong\u003e\u003cp\u003e\u003cem\u003ePunica granatum\u003c/em\u003e\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eFamily\u003c/b\u003e: Lythraceae, subfamily: Punicoideae\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eGenus\u003c/strong\u003e\u003cp\u003ePunica\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eKingdom\u003c/strong\u003e\u003cp\u003ePlantae\u003c/p\u003e\u003c/p\u003e\u003cp\u003eThe mature fresh fruit peels of pomegranate fruits (\u003cem\u003ePunica granatum\u003c/em\u003e) were gathered at the Eluru, Andhra Pradesh fruit market in January 2025, and Dr. Y.S.R. Horticultural University, AP, India's fruit science department verified Bhagwa (NRCP H-12). The National Collection of Industrial Microorganisms (NCIM) National Chemical Laboratory, Pune, and the Microbiology Laboratory, Sri Vasavi Institute of Pharmaceutical Sciences, provided the Propionibacterium acnes and Staphylococcus aureus tested microorganisms utilized in this investigation. Mueller Hinton Agar was utilized as the growth medium (MHA-OXOID). Amyl alcohol, 10% ammonia, 2N hydrochloric acid, iron (III) chloride, ether, chloroform, anhydrous acetic acid solution in concentrated H2SO4, 1% gelatin, and the reagent Dragendorff (potassium bismuth iodide solution) were among the substances utilized. sterile physiological sodium chloride, ethanol, carbopol, propylene glycol, methylparaben/propylparaben, triethanolamine, 10% vanillin solution in concentrated H2SO4, 1N sodium hydroxide, potassium permanganate powder, magnesium powder, and Mayer reagents (potassium mercury iodide solution). All of Yucca Enterprises' chemicals and Nava Bharat Ventures Ltd.'s alcohol (AAA CN 7327 CXM 005).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eChemical Constituents present in Punica granatum Peel\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"2\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eChemical Constituents\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eUses\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGallic acid\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAnti-inflammatory, Antimicrobial, Antioxidant.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eEllagic acid\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eInhibits acne-causing bacteria, Anti-inflammatory, Inhibits sebum accumulation.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePunicalin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAntiviral, Antifungal, Antioxidant.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePunicalagin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAntioxidant, Anti-inflammatory, Antiproliferative, Anti-angiogenic, Hepatoprotective, Anti-atherosclerotic, Chemo preventive, Reduces skin pigmentation, Antidepressant.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCaffeic acid\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAntibacterial, Anti-inflammatory, Antioxidant.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eEllagitannins\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAntibacterial, Anti-inflammatory, Skin barrier protection.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePelletierine alkaloids\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eVermicide, Anti-cancerous.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLuteolin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAntimicrobial, Antiviral, Anti-inflammatory, Antioxidant, Wound healing.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKaempferol\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAntimicrobial, Anti-inflammatory, Inhibits excess sebum, Anti-cancer, Antioxidant.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eQuercetin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAntimicrobial, Anti-inflammatory, Reduces skin erythema.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e[\u003cspan additionalcitationids=\"CR12\" citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e[\u003cspan additionalcitationids=\"CR15\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003ePreparation of Extract\u003c/h3\u003e\n\u003cp\u003eAfter washing and slicing the fruits, the arils and carpelar membranes were manually extracted from the fruit skins. Sliced and dried in a convection oven at 50\u0026deg;C for three days, the peels were then crushed in an electric coffee grinder. After weighing the resultant powders, the findings were recorded. To extract the antibacterial components of pomegranate peel, the following crude extract and its separated fractions were created. For extraction, 300 g of peel powder from each pomegranate cultivar was macerated in 1 L of 70% ethanol, the maceration liquid was allowed to sit at room temperature for 72 hours in the dark. Following their passage through Whatman filter paper No. 1 (Labsman Filtration System, India), the extracts were then filtered through a micropore glass filter. The crude extracts, or ethanolic extracts, were evaporated at 40\u0026deg;C with reduced pressure in a rotary evaporator until it turns into a thick slurry. Entire Process is illustrated in the Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e1\u003c/span\u003e Schematic representation of the pomegranate peel extraction process.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\n\u003ch3\u003ePhytochemical screening of the extract\u003c/h3\u003e\n\u003cp\u003eA phytochemical screening of plants was conducted to identify the secondary metabolite group present in the pomegranate peel (\u003cem\u003ePunica granatum\u003c/em\u003e) extract. Polyphenols, tannins, flavonoids, and phenolic acids are some of these metabolites.\u003c/p\u003e\n\u003ch3\u003eFormulation of Pomegranate Peel Extract Gel\u003c/h3\u003e\n\u003cp\u003eAccurately, a weighed quantity of polymers Carbopol 940 (1% w/v) was distributed in purified water and allowed to swell overnight. After swelling of the polymer, the penetration enhancer (Di sodium EDTA), glycerin, and Pomegranate Peel Extract along with aloe vera moisturizer were combined to the solution with stirring and buffer solution, preservatives solution, and essential oils were also added. The final volume was made up to 100ml with purified distilled water and manufactured gels were stored in a bottle with proper labeling (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eFormulation of Pomegranate Peel Extract Gel\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"6\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eIngredient\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eFunction\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eConcentration (% w/w)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eLow Strength (0.5%)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eMedium Strength (1.0%)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eHigh Strength (2.0%)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eFormula 1\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eFormula 2\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eFormula 3\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003ePomegranate Peel Extract\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eActive ingredient\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.5\u0026ndash;2.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e2.0\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCarbomer 940\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGelling agent\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.5\u0026ndash;1.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.75\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.0\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eGlycerin\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHumectant\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e3.0\u0026ndash;5.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e5.0\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eAloe Vera Gel\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMoisturizer, soother\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e10.0\u0026ndash;15.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e10.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e15.0\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003ePhenoxyethanol\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePreservative\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.5\u0026ndash;1.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.75\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.0\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eDi sodium EDTA\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePreservative\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.1\u0026ndash;0.2% (100\u0026ndash;200 mg)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eBuffer solution\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003epH adjuster (neutralizer)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eq.s. to pH 5.5\u0026ndash;6.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eq.s.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eq.s.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eq.s.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eRose Oil (optional)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eFragrance\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.05\u0026ndash;0.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eDistilled Water\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eDiluent\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eUp to 100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eq.s.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eq.s.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eq.s.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\n\u003ch3\u003eDosage Strength Variations\u003c/h3\u003e\n\u003cp\u003e\u003cul\u003e\u003cli\u003e\u003cp\u003e\u003cb\u003eLow Strength (0.5%)\u003c/b\u003e: Suitable for sensitive skin or as a preventive skincare product.\u003c/p\u003e\u003c/li\u003e\u003cli\u003e\u003cp\u003e\u003cb\u003eMedium Strength (1.0%)\u003c/b\u003e: Moderate dosage for general skin enhancement.\u003c/p\u003e\u003c/li\u003e\u003cli\u003e\u003cp\u003e\u003cb\u003eHigh Strength (2.0%)\u003c/b\u003e: High potency for therapeutic or targeted benefits.\u003c/p\u003e\u003c/li\u003e\u003c/ul\u003e\u003c/p\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eEvaluation of formulations\u003c/h2\u003e\u003cdiv id=\"Sec9\" class=\"Section3\"\u003e\u003ch2\u003ePhysical evaluation\u003c/h2\u003e\u003cp\u003eColour, look, and consistency were among the physical characteristics that were visually examined.\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\n\u003ch3\u003eWashability\u003c/h3\u003e\n\u003cp\u003eFollowing the application of the formulations to a skin-like substrate, a manual assessment was carried out to evaluate the ease and thoroughness of removal using water.\u003c/p\u003e\u003cp\u003e\u003cb\u003epH\u003c/b\u003e\u003c/p\u003e\u003cp\u003eA pH test was performed on a 1% aqueous solution of the formulation using a calibrated digital pH meter that was kept at a constant temperature.\u003c/p\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003eSpreadability\u003c/h2\u003e\u003cp\u003eAn internally developed and planned gadget was used to measure spreadability. With a fixed glass slide and a movable glass slide, the gadget is constructed from a hardwood block. On a pulley that was horizontally level with the fixed slide, one end of the moveable glass slide was fastened to a weight pan that was rolled. The produced gel's spreadability was measured using its \"Slip and Drag\" characteristics. The surplus gel from the trial (about 2g) was placed on this ground slide. After that, two slides were placed in between the gel. A one-kilogram weight was placed on top of each slide for five minutes to push air out and form a uniform gel layer between them. The borders of the extra gel were scraped off. After that, the top plate was given a 50 gramme pull-off. Use the thread attached to the hook to mix, and note the time it takes for the top slide to move 7.5 cm (T, in seconds). Shorter intervals were better for spreadability.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003eStability study\u003c/h2\u003e\u003cp\u003eOut of all the formulations that are generated, stability research is done to find the one that has the most drug release and may thus be referred to as the \"best formulation.\" For one month, a stability study was conducted; the formulation was kept at 40\u0026deg;C, 75% relative humidity, and 40\u0026deg;C in a stability chamber. The formulation's properties, including phase separation pH and drug content, were assessed after a month.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003eAnti-Bacterial Activities against P. acnes and S. aureus\u003c/h2\u003e\u003cp\u003e\u003cem\u003eCutibacterium acnes\u003c/em\u003e (formerly \u003cem\u003ePropionibacterium acnes\u003c/em\u003e) is a Gram-positive, anaerobic bacterium that plays a significant role in acne vulgaris. It contributes to the condition by producing lipases that degrade sebum into free fatty acids, leading to inflammation and blockage of pilosebaceous units[\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. \u003cem\u003eStaphylococcus aureus\u003c/em\u003e, another Gram-positive bacterium commonly found on human skin, has also been investigated for its potential involvement in acne. However, studies have shown no significant difference in \u003cem\u003eS. aureus\u003c/em\u003e colonization rates between acne patients and healthy individuals, suggesting it may not play a central role in acne pathogenesis[\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. P. acnes (NCIM 0266) and S. aureus (NCIM 2654) were provided for the study by the National Collection of Industrial Microorganisms (NCIM) at the National Chemical Laboratory in Pune. Mueller Hinton Agar (MHA-OXOID) was used to cultivate P. acnes in an anaerobic environment that was made with MGC AnaeroPack-Anaero and MGC AnaeroPack Jar. An Agar-Agar media was used to culture S. aureus. The paper disc-diffusion method was employed to evaluate the efficacy of Pomegranate peel extract against P. acnes and S. aureus. A 10ml suspension of P. acnes (4 x 10\u003csup\u003e8\u003c/sup\u003e CFU/mL) was introduced into 90mL of sterile media at 45\u0026deg;C in a water bath under aseptic conditions. Immediately after inoculation, the agar media were combined and moved to a petri plate. To get concentrations between 0.5% and 2%, the pomegranate peel extract gel was taken in sterile distilled water in accordance with the formulation strength. Distilled ethanol served as the negative control, while clindamycin (10 U) served as the positive control. Millimeters were used to measure the zones of inhibition after a 24-hour anaerobic incubation period at 37\u0026deg;C. The anti-S. aureus (0.5 x 108 CFU/mL) activity evaluation technique was slightly altered. The minimum bacterial concentration (MBC) of the sample was found to be the lowest value that prevented detectable bacterial growth. It was found that the minimum inhibitory concentration, or MIC, was the lowest sample concentration at which observable growth occurred.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003ch2\u003eUV-Vis Spectral Analysis\u003c/h2\u003e\u003cp\u003eThe pomegranate peel extract was dissolved in ethanol at a concentration of 1\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\mu\\:g/ml\\)\u003c/span\u003e\u003c/span\u003e. The solution was scanned in the UV-Vis spectrophotometer between 190\u0026ndash;800 nm using a quartz cuvette with methanol as a blank. The wavelength at which maximum absorption \u003cb\u003e(\u003c/b\u003eλ max\u003cb\u003e)\u003c/b\u003e occurred was recorded to identify the presence of bioactive compounds.\u003c/p\u003e\u003cp\u003e\u003cb\u003eIn vitro\u003c/b\u003e \u003cb\u003eantioxidant activity\u003c/b\u003e\u003c/p\u003e\u003cp\u003e2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging test[\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Standard ascorbic acid and sample were prepared in methanol in different concentrations: 25, 50, 100, 200, 500, and 1000 \u0026micro;g/ml. The free radical used was 0.1 mM DPPH in methanol. Each test tube was cleaned, labelled, and filled with an equal amount of DPPH and standards at various concentrations. The control was similarly filled with an equal volume of methanol and DPPH, and the tubes were left to sit at room temperature in the dark for half an hour. The absorbance was measured with a UV-Vis spectrophotometer at 517 nm. The degree of continuous DPPH decolorization from purple to DPPHH yellow demonstrated how efficient the sample was at scavenging. Using the following formula, the sample's scavenging activity against the stable DPPH was determined:\u003c/p\u003e\u003cp\u003eScavenging activity (%) = \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\left(\\frac{A-B}{A}\\right)\\times\\:100\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\u003cp\u003eWhere A is the absorbance of the control and\u003c/p\u003e\u003cp\u003eB is the absorbance of the sample/standard\u003c/p\u003e\u003cp\u003e\u003cb\u003eIn vitro\u003c/b\u003e \u003cb\u003eanti-inflammatory activity\u003c/b\u003e\u003c/p\u003e\u003cp\u003eCollect the goat blood into a sterile air-tight container with previously added anti-coagulant like EDTA and store below 20\u003csup\u003eo\u003c/sup\u003eC (Don\u0026rsquo;t freeze). Take the whole blood into the centrifuge tubes and centrifuge at 3000rpm for 10min, remove the supernatant fluid from the tubes and wash by adding normal saline (0.9%) and centrifuge, decant the supernatant fluid and repeat this for 2 more times. Resuspend RBC in the normal saline (0.9%) to get 10% RBC suspension. Prepare the pomegranate peel extract sample with serial dilution into 25, 50, 100, 200, 400, 500, 800, and 1000\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\mu\\:g/ml\\)\u003c/span\u003e\u003c/span\u003e. In the same way, prepare the standard dilutions of drug Diclofenac into 200\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\mu\\:g/ml\\)\u003c/span\u003e\u003c/span\u003e.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\u003ch2\u003eHypotonicity-Induced Hemolysis Method\u003c/h2\u003e\u003cp\u003eIn sterile test tubes add an equal quantity of 10%RBC suspension and 1mL sample of each dilution and label them accordingly, also prepare the standard tubes in the same way along with the control tube by adding an equal quantity of 10% RBC suspension and hypotonic solution (50mM NaCl solution) and incubate all at the 37\u003csup\u003eo\u003c/sup\u003eC for 10 min.\u003c/p\u003e\u003cp\u003eAfter incubation centrifuge above the sample at 3000rpm for 10 min collect the supernatant fluid measure the absorbance at 540nm in the UV-Vis spectrophotometer and take down the absorbance readings. Calculate the % inhibition and plot the graph between the %inhibition and concentration.\u003c/p\u003e\u003cp\u003e%Inhibition = \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{Absorbance\\:of\\:Control-Absorbance\\:of\\:Sample\\:or\\:Reference}{Absorbance\\:of\\:Control}\\)\u003c/span\u003e\u003c/span\u003e \u0026times; 100\u003c/p\u003e\u003cp\u003eThe higher the %inhibition the better the anti-inflammatory activity and vice versa.\u003c/p\u003e\u003c/div\u003e"},{"header":"RESULTS","content":"\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\u003ch2\u003eRate of Extraction\u003c/h2\u003e\u003cp\u003eFrom the 100 grams, 150 grams, and 300 grams of the dried pomegranate peels total of 20%, 32.66%, and 53.33% respectively was obtained (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The extract was dried in a hot air oven and stored in an ambered colored air-tight container away from the sunlight at 2-8\u003csup\u003eo\u003c/sup\u003ec. For the further analysis.\u003c/p\u003e\u003cp\u003e%purity equation = \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{Crude\\:Drug}{Extract}\\times\\:100\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eRate of Extraction from peels\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCrude Drug\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eExtract Weight\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e% Purity\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTrail 1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e100gm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e20gms\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e20%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTrail 2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e150gm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e49gms\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e32.66%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTrail 3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e300gm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e160gm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e53.33%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\u003ch2\u003ePhytochemical screening\u003c/h2\u003e\u003cp\u003eThe phytochemical screening of the extract revealed the presence of phenolic compounds, flavonoids, tannins, alkaloids, terpenoids, steroids, carbohydrates, proteins, and saponins. However, glycosides and volatile oils were absent as shown in the Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003ePhotochemical Screening of Pomegranate Peel Extract\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eConstituent\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eTest\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eProcedure\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eExpected Result\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eTest Result\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePhenolic Compounds\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eFerric chloride test\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAdd ferric chloride to the extract.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eBlue-black/green color\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePositive\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFlavonoids\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eShinoda test\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAdd magnesium turnings along with\u003c/p\u003e\u003cp\u003econcentrated HCl to the extract.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ePink/red/orange color\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePositive\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTannins\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGelatin test\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAdd 1% gelatin solution with NaCl to the extract.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eWhite precipitate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePositive\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAlkaloids\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eDragendorff\u0026rsquo;s test\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAdd Dragendorff\u0026rsquo;s reagent to the extract.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eReddish-brown precipitate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePositive\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGlycosides\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eKeller-Kiliani test\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAdd glacial acetic acid, ferric chloride, and sulfuric acid to the extract.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eReddish-brown ring at the interface\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eNegative\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTerpenoids\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSalkowski test\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAdd concentrated sulfuric acid to the extract.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eReddish-brown interface\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePositive\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSaponins\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eFroth test\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eShake the extract with water vigorously.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ePersistent froth\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePositive\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eVolatile Oils\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eFilter paper test\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePress the extract on filter paper.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eAromatic scent and translucent\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eNegative\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSteroids\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLiebermann-Burchard test\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAdd acetic anhydride and concentrated sulfuric acid to the extract.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eGreen/blue coloration\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePositive\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCarbohydrates\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMolisch test\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAdd Molisch reagent and concentrated sulfuric acid to the extract.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eViolet ring formation\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePositive\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eProteins\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBiuret test\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAdd sodium hydroxide and copper sulfate to the extract.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eViolet coloration\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePositive\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e\u003ch2\u003eEvaluation of Formulation\u003c/h2\u003e\u003cdiv id=\"Sec20\" class=\"Section3\"\u003e\u003ch2\u003ePhysical Evaluation\u003c/h2\u003e\u003cp\u003e\u003cul\u003e\u003cli\u003e\u003cp\u003e\u003cb\u003eColor\u003c/b\u003e: Reddish-brown, uniform throughout.\u003c/p\u003e\u003c/li\u003e\u003cli\u003e\u003cp\u003e\u003cb\u003eAppearance\u003c/b\u003e: Smooth, homogenous gel with no visible lumps or phase separation.\u003c/p\u003e\u003c/li\u003e\u003cli\u003e\u003cp\u003e\u003cb\u003eConsistency\u003c/b\u003e: Semi-solid, non-greasy, and easy to spread\u003c/p\u003e\u003c/li\u003e\u003c/ul\u003e\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec21\" class=\"Section2\"\u003e\u003ch2\u003eWashability\u003c/h2\u003e\u003cp\u003e\u003cul\u003e\u003cli\u003e\u003cp\u003eThe formulation was easily washable with water, leaving no sticky or oily residue on the skin.\u003c/p\u003e\u003c/li\u003e\u003c/ul\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec22\" class=\"Section2\"\u003e\u003ch2\u003epH Measurement\u003c/h2\u003e\u003cp\u003e\u003cul\u003e\u003cli\u003e\u003cp\u003eThe pH of the 1% aqueous solution was found to be 5.5 to 6.5, which is within the ideal range for skin applications and matches the natural skin pH.\u003c/p\u003e\u003c/li\u003e\u003c/ul\u003e\u003c/p\u003e\u003cdiv id=\"Sec23\" class=\"Section3\"\u003e\u003ch2\u003eSpreadability\u003c/h2\u003e\u003cp\u003e\u003cul\u003e\u003cli\u003e\u003cp\u003eThe spreadability time was recorded as less than 10 seconds, indicating excellent spreadability and ease of application.\u003c/p\u003e\u003c/li\u003e\u003c/ul\u003e\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec24\" class=\"Section2\"\u003e\u003ch2\u003eStability Study\u003c/h2\u003e\u003cp\u003e\u003cul\u003e\u003cli\u003e\u003cp\u003e\u003cb\u003ePhase separation\u003c/b\u003e: No phase separation observed. The gel remained uniform.\u003c/p\u003e\u003c/li\u003e\u003cli\u003e\u003cp\u003e\u003cb\u003epH stability\u003c/b\u003e: No significant change in pH, maintaining a stable value between 5.5 to 6.5.\u003c/p\u003e\u003c/li\u003e\u003cli\u003e\u003cp\u003e\u003cb\u003eDrug content\u003c/b\u003e: No significant degradation; the pomegranate peel extract retained over 95% of its initial active compounds.\u003c/p\u003e\u003c/li\u003e\u003c/ul\u003e\u003c/p\u003e\u003cp\u003eThe formulation demonstrated good stability, ease of application, and compatibility with the skin, making it a promising topical preparation.\u003c/p\u003e\u003cp\u003e\u003cb\u003eAnti-Bacterial Activities against\u003c/b\u003e \u003cb\u003eP. acnes\u003c/b\u003e \u003cb\u003eand\u003c/b\u003e \u003cb\u003eS. aureus\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eAnti-bacterial activity of Pomegranate Peel Extract\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eSample\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e\u003cp\u003eAnti-bacterial activity\u003c/p\u003e\u003cp\u003e[zone of inhibition {mm}]\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePropionibacterium\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eStaphylococcus epidermidis\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eFormula 1 (0.5%)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eFormula 2 (1%)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eFormula 3 (2%)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eDistilled Ethanol\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eClindamycin\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eTo tackle microbial infections, researchers are attempting to get novel elements from natural resources due to the rising occurrence of multi-drug resistance in microbes globally[\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. The plants undoubtedly contain a wealth of bioactive compounds with significant therapeutic potential. This study examined \u003cem\u003ePunica granatum\u003c/em\u003e peel extract's anti-acne properties against P. acnes and S. aureus as trends observed in Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e. According to antimicrobial activity studies, the extract from the gel of pomegranate peels from \u003cem\u003ePunica granatum\u003c/em\u003e showed a high (24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31) zone of inhibition against Staphylococcus aureus and Propionibacterium acnes. This study shows that the inhibition zone of the gel containing pomegranate peel extract is equivalent to that of regular clindamycin. Phenols, flavonoids, saponins, alkaloids, and tannins (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) are among the phytochemicals that provide Pomegranate Peel Extract gel its anti-acne properties. Phenols interfere with the cytoplasmic membrane's normal function, which impacts the production of nucleic acids[\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Alkaloids and terpenoids interact with the proteins and enzymes in the microbial cell membrane, disrupting it and releasing a flow of protons outside the cell, which can either cause cell death or inhibit enzymes essential for the manufacture of amino acids[\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. RNA polymerase, reverse transcriptase, telomerase, and DNA polymerase are bacterial enzymes that flavonoids have been shown to inhibit. The saponins induce intracellular molecules to flow out by lowering surface tension, which promotes cell permeability or leakage[\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e\u003cdiv id=\"Sec25\" class=\"Section3\"\u003e\u003ch2\u003eUV-Vis Spectral Analysis\u003c/h2\u003e\u003cp\u003ePomegranate peel extract's UV-Vis spectra showed distinctive peaks in the 190\u0026ndash;800 nm region, suggesting the presence of flavonoids and phenolic chemicals. A prominent absorption peak, which corresponds to phenolic acids like gallic and ellagic acids, was seen at 250\u0026ndash;280 nm in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The 350\u0026ndash;400 nm range showed another notable peak, indicating the presence of flavonoids such as flavonols and anthocyanins. Pomegranate peels contain polyphenolic chemicals; they are widely recognized for their strong anti-inflammatory and antioxidant properties. The absorption pattern is consistent with other studies on these substances. These bioactive substances lend support to the extract's possible pharmaceutical uses, such as membrane stabilization in tests for anti-inflammatory responses. The wide UV absorption bands attest to the existence of aromatic rings and Phenolics have conjugated double bonds. These results are in line with other studies that showed flavonoids, punicalagin, and ellagitannins as the main components of pomegranate peel extract.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eIn vitro\u003c/b\u003e \u003cb\u003eantioxidant activity\u003c/b\u003e\u003c/p\u003e\u003cp\u003eDPPH scavenging of free radicals. The Table\u0026nbsp;\u003cspan refid=\"Tab7\" class=\"InternalRef\"\u003e7\u003c/span\u003e displays the antioxidant activity results for various sample concentrations and reference ascorbic acid. Through the scavenging of DPPH (free radical) and its conversion to DPPHH, the samples demonstrated significant antioxidant activity. Radical scavenging activity was shown to be dose-dependent. All six extract concentrations had lower scavenging efficacy than ascorbic acid. When an antioxidant is present, the DPPH radical gains an additional electron and the absorbance falls.\u003c/p\u003e\u003cp\u003eAccording to this study, pomegranate peel extract's scavenging activity was dose-dependent, meaning that the higher the concentration, the greater the scavenging activity. Even though the samples Ascorbic acid and pomegranate peel extract had almost the same DPPH radical scavenging activity based on the results from Table\u0026nbsp;\u003cspan refid=\"Tab7\" class=\"InternalRef\"\u003e7\u003c/span\u003e Anti-oxidant activity of Pomegranate Peel Extract. The study demonstrated that pomegranate peel extract may donate protons and may function as a main antioxidant by scavenging or suppressing free radicals.\u003c/p\u003e\u003cp\u003eAscorbic acid was shown to have an IC₅₀ value between 50 and 100 \u0026micro;g/ml, whereas pomegranate peel extract showed 50% inhibition between 100 and 200 \u0026micro;g/ml, indicating a moderate antioxidant potential based on the graphical representation in Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e3\u003c/span\u003e.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe antioxidant activity of the pomegranate peel anti-acne gel formulations showed a statistically significant difference (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) among the different concentrations/treatments, as determined by one-way ANOVA, indicating a concentration-dependent increase in antioxidant potential.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab7\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eAnti-oxidant activity of Pomegranate Peel Extract\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eConcentration\u003c/p\u003e\u003cp\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:(\\varvec{\\mu\\:}\\varvec{g}/\\varvec{m}\\varvec{l})\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003e% Inhibition of Ascorbic acid\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e% Inhibition of Pomegranate\u003c/p\u003e\u003cp\u003ePeel Extract\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e41.16424116\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e12.47401247\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e46.56964657\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e29.93762994\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e100\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e65.28066528\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e46.36174636\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e200\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e68.3991684\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e91.26819127\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e500\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e93.34719335\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e92.72349272\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1000\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e95.42619543\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e92.93139293\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eIn vitro\u003c/b\u003e \u003cb\u003eanti-inflammatory activity\u003c/b\u003e\u003c/p\u003e\u003cp\u003ePomegranate peel extract's anti-inflammatory properties were assessed by the hypotonicity-induced hemolysis test. Calculated for various extract concentrations, the percentage inhibition of hemolysis was compared to that of the standard medication, Diclofenac. The findings show that the extract protects against RBC membrane lysis in a concentration-dependent manner. Increased inhibition was seen at higher extract doses, indicating strong membrane stabilization action. To ascertain how well the extract prevented hypotonicity-induced hemolysis, the absorbance values at 540 nm and the associated percentage inhibition were examined and reported in the Table\u0026nbsp;\u003cspan refid=\"Tab8\" class=\"InternalRef\"\u003e8\u003c/span\u003e Anti-inflammatory activity of Pomegranate Peel Extract. To show the extract's effectiveness in comparison to Diclofenac, a percent inhibition vs concentration graph was created.\u003c/p\u003e\u003cp\u003eIn vitro anti-inflammatory activity of Pomegranate peel extract was concentration-dependent, the maximum protection of 74.23% was seen at the concentration of 1000\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\mu\\:g\\)\u003c/span\u003e\u003c/span\u003e/ml. All results were compared with reference diclofenac sodium which showed 73.98 protection at the concentration of 1000\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\mu\\:g\\)\u003c/span\u003e\u003c/span\u003e/ml as shown in the Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e4\u003c/span\u003e.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe anti-inflammatory activity of the pomegranate peel anti-acne gel exhibited statistically significant variation among the tested formulations (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05), as determined by one-way ANOVA, suggesting that increased concentrations of the extract enhance the formulation\u0026rsquo;s anti-inflammatory efficacy.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab8\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 8\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eAnti-inflammatory activity of Pomegranate Peel Extract.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eConc (\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\varvec{\\mu\\:}\\varvec{g}/\\varvec{m}\\varvec{l})\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003e% Inhibition of Pomegranate peel extract\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e% Inhibition of Diclofenac sodium\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e6.89\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e5.75\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e10.75\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e9.24\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e100\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e15.79\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e13.69\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e200\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e18.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e17.26\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e400\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e31.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e29.45\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e500\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e34.62\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e35.78\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e800\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e61.42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e59.12\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1000\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e74.23\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e73.98\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe results of this investigation show the medicinal potential of pomegranate peel extract (\u003cem\u003ePunica granatum\u003c/em\u003e), namely in anti-inflammatory, antibacterial, and antioxidant applications. Significant bioactive substances were confirmed by phytochemical screening, which was supported by the high extraction yield.\u003c/p\u003e\u003cdiv id=\"Sec27\" class=\"Section2\"\u003e\u003ch2\u003eComparison with Existing Research\u003c/h2\u003e\u003cp\u003eThe extract had vigorous antibacterial activity against Propionibacterium acnes and Staphylococcus aureus, with inhibition zones that were like those of clindamycin, a popular antibiotic used to treat acne. This supports earlier research showing the antibacterial activity of polyphenols, flavonoids, tannins, and alkaloids produced from pomegranates, which damage bacterial membranes, prevent the formation of proteins, and interfere with metabolic processes. These substances likely work in concert to strengthen the antibacterial action.\u003c/p\u003e\u003cp\u003eWhen evaluated using the DPPH radical scavenging assay, the antioxidant qualities showed dose-dependent action, with higher doses producing more scavenging potential. The extract's capacity to neutralize radicals was somewhat less than that of ascorbic acid, but still rather significant. These findings suggest the extract's potential use in dermatological applications, as oxidative stress is linked to inflammatory disorders and skin aging. In keeping with earlier research showing the robust ability of pomegranate peel extracts to scavenge free radicals, the presence of phenolic acids (gallic and ellagic acids), flavanols, and anthocyanins, as verified by UV-Vis spectral analysis, further supports its antioxidant potential.\u003c/p\u003e\u003cp\u003eSignificant membrane-stabilizing effects like those of diclofenac sodium were established by the anti-inflammatory action, as assessed by hypotonicity-induced hemolysis inhibition. This implies that the extract from pomegranate peels decreases erythrocyte lysis, which is like how biological membranes have anti-inflammatory properties. The extract is efficient in reducing inflammation, maybe by stabilizing lysosomal membranes and inhibiting pro-inflammatory mediators, as evidenced by its high inhibition percentage at higher doses (74.23% at 1000 \u0026micro;g/ml). These findings are consistent with earlier studies showing that pomegranate polyphenols affect oxidative stress-related inflammation pathways and reduce inflammatory cytokines.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec28\" class=\"Section2\"\u003e\u003ch2\u003eHPTLC Analysis of Pomegranate Peel Extract\u003c/h2\u003e\u003cp\u003eTo ensure the quality and consistency of the pomegranate peel extract, High-Performance Thin-Layer Chromatography (HPTLC) analysis was conducted. The hydroalcoholic extract of \u003cem\u003ePunica granatum\u003c/em\u003e peel was subjected to HPTLC fingerprinting using a CAMAG HPTLC system. Pre-coated silica gel 60 F254 plates were used as the stationary phase. The sample was applied in bands using a CAMAG Linomat V applicator, and the mobile phase consisted of toluene:ethyl acetate:formic acid (5:4:1, v/v/v), which provided optimal resolution of phenolic and flavonoid compounds. After development, the plates were dried and scanned at 254 nm and 366 nm using a CAMAG TLC Scanner III. Distinct bands corresponding to gallic acid, ellagic acid, and punicalagin were observed and confirmed by comparing Rf values with standard reference compounds. This chromatographic profiling confirms the presence of major bioactive constituents and can serve as a reliable tool for quality control of the extract in future formulations.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec29\" class=\"Section2\"\u003e\u003ch2\u003eImplications of Findings\u003c/h2\u003e\u003cp\u003eIn formulations that address inflammation, oxidative stress, and acne, the results indicate that pomegranate peel extract can be used as a natural, plant-based substitute. The development of herbal-based antimicrobial remedies is becoming increasingly popular due to the growth in antibiotic resistance. Given its effectiveness against P. acnes and S. aureus, the extract may find application in formulations for acne therapy, either by itself or in conjunction with more traditional agents. Additionally, its antioxidant qualities could support formulas that protect the skin and fight aging. Its anti-inflammatory properties make it an important element in pharmaceutical and dermatological formulations, since they may help soothe irritated or inflamed skin.\u003c/p\u003e\u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003e\u003cem\u003ePunica granatum\u003c/em\u003e (pomegranate) peel extract, which is made via hydroalcoholic maceration and formed into a topical gel, has been shown in this study to have strong anti-acne properties. Strong antibacterial activity against Propionibacterium acnes and Staphylococcus aureus was demonstrated by UV-Vis spectral analysis, and this was attributed to the extract's high phenolic and flavonoid content. Furthermore, the extract demonstrated significant anti-inflammatory and antioxidant qualities, confirming its applicability in treating acne.\u003c/p\u003e\u003cp\u003eThe developed gel presents a viable natural acne treatment option with potential benefits including biocompatibility, less adverse effects than synthetic therapies, and multipurpose skin benefits. To compare its clinical effectiveness, stability, and skin permeability to traditional acne therapies, more research is necessary. In vivo trials, formulation optimization, and long-term safety evaluations should all be investigated in future studies to confirm its therapeutic potential.\u003c/p\u003e\u003cp\u003eThese results highlight the potential of pomegranate peel extract as a useful component in dermatological formulations, opening the door for its use in environmentally friendly, plant-based acne remedies.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAbbreviations\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical Approval\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study did not involve any human or animal participants that would require formal ethical approval. The formulation development and in vitro evaluations only were conducted following standard pharmaceutical research protocols.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics and Consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe mature fresh fruit peels of pomegranate (\u003cem\u003ePunica granatum\u003c/em\u003e, cultivar Bhagwa [NRCP H-12]) were gathered in January 2025 from the Eluru fruit market, Andhra Pradesh, India. The plant material was taxonomically authenticated by the Department of Fruit Science, Dr. Y.S.R. Horticultural University, Andhra Pradesh, India. The pomegranate peels were obtained from cultivated fruits that are widely grown and marketed in India; hence, no wild collection or endangered plant material was involved. The study complied with local and national guidelines for the use of plant materials in research. Since the fruits were procured from a commercial market and did not involve endangered or protected species, no special permissions or licenses were required.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for Publish\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors consent to the publication of this work in its current form, following peer review and editorial revisions.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of Data and Materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data supporting this study are available upon request from the corresponding author.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNo Funding Received\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical Trial Registration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable. This study did not involve any clinical trial.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical Trial Number\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cul type=\"disc\"\u003e\n \u003cli\u003e\u003cstrong\u003eNeelam Adhi Kesava Naidu\u003c/strong\u003e: Conducted the research, performed experiments, analyzed data, and drafted the manuscript.\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eJ.N.B. Indusekhar\u003c/strong\u003e: Provided supervision, technical guidance, and constructive feedback throughout the research process.\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eDr. V. Bhaskara Raju\u003c/strong\u003e: Granted the necessary permissions and provided administrative support for the research.\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003eAll authors have \u003cstrong\u003ereviewed and approved\u003c/strong\u003e the final manuscript and agree to be accountable for their respective contributions.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors express their sincere gratitude to Sri Vasavi Institute of Pharmaceutical Sciences for providing the necessary facilities and support to carry out this research. A special thanks to \u003cstrong\u003eJ.N.B. Indusekhar,\u003c/strong\u003e whose valuable guidance and expertise greatly contributed to the successful completion of this study. We also extend our appreciation to \u003cstrong\u003eDr.V. Bhaskara Raju\u003c/strong\u003e, Principal of Sri Vasavi Institute of Pharmaceutical Sciences, for granting the required permissions and institutional support. Finally, the corresponding author, \u003cstrong\u003eNeelam\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eAdhi Kesava Naidu\u003c/strong\u003e, acknowledges the tireless efforts dedicated to conducting the research and preparing this manuscript. No additional contributors or external support were involved in the preparation of this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompliance with Guidelines for Studies Involving Plants\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study involved the use of \u003cstrong\u003epomegranate (\u003cem\u003ePunica granatum\u003c/em\u003e) peels\u003c/strong\u003e\u003cstrong\u003e,\u003c/strong\u003e which were collected from local sources in \u003cstrong\u003eAndhra Pradesh, India\u003c/strong\u003e\u003cstrong\u003e.\u0026nbsp;\u003c/strong\u003eThe study complied with all applicable \u003cstrong\u003elocal, national, and international guidelines and regulations\u003c/strong\u003e for plant research. As the research did not involve endangered or protected plant species and only utilized \u003cstrong\u003eagricultural by-products (peels) from commercially available pomegranates\u003c/strong\u003e\u003cstrong\u003e,\u0026nbsp;\u003c/strong\u003eno specific permits or ethical approvals were required. The handling and processing of plant material were conducted following \u003cstrong\u003estandard phytochemical and pharmaceutical research protocols\u003c/strong\u003e toensure scientific integrity and reproducibility.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data supporting this study are available upon request from the corresponding author.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eC.E.M.J.E.B.P.Y.L.C.G.S. Draelos ZD, United States/Canada Dapsone Gel Study Group. Two randomized studies demonstrate the efficacy and safety of dapsone gel, 5% for the treatment of acne vulgaris. , J Am Acad Dermatol 56 (2007) 439. https://doi.org/http://dx.doi.org/10.1016/j.jaad.2006.10.005.\u003c/li\u003e\n \u003cli\u003eM.K.L.C. Wainwright M, Phenothiazinium photosensitisers IX. Tetra-and pentacyclic derivatives as photoantimicrobial agents, Dyes and Pigments 91 (2011) 1\u0026ndash;5.\u003c/li\u003e\n \u003cli\u003eR.R.B.A.W.D.S.D.G.R.N.M.W.J. Berger R, Tretinoin gel microspheres 0.04% versus 0.1% in adolescents and adults with mild to moderate acne vulgaris: a 12-week, multicenter, randomized, double-blind, parallel-group, phase IV trial , Clin Ther 29 (2007) 1086\u0026ndash;1097.\u003c/li\u003e\n \u003cli\u003eA.J.H.G.P.D.S.W. Magin P, Psychological sequelae of acne vulgaris: results of a qualitative study, Canadian Family Physician 52 (2006) 978\u0026ndash;979.\u003c/li\u003e\n \u003cli\u003eS.JC. Haider A, Treatment of acne vulgaris, JAMA 296 (2004) 726\u0026ndash;735.\u003c/li\u003e\n \u003cli\u003eK.D.L.J.L.A.S.A.S.E.T.D.V.V.A.B.K.S.C.B.R. Strauss JS, Guidelines of care for acne vulgaris management, J Am Acad Dermatol 56 (2007) 651\u0026ndash;663.\u003c/li\u003e\n \u003cli\u003eG.R.K.S.G.GD. Rashmi MS, Topical gel: A review, Pharm Rev 6 (2008) 1\u0026ndash;3.\u003c/li\u003e\n \u003cli\u003eM.A.H.A.J.A. Ramli R, Acne analysis, grading and computational assessment methods: an overview, Skin Research and Technology 18 (2012) 1\u0026ndash;4.\u003c/li\u003e\n \u003cli\u003eH.K.N.P. Plewig G, Clinical and bacteriological evaluation of nadifloxacin 1% cream in patients with acne vulgaris: a double-blind, phase III comparison study versus erythromycin 2% cream, European Journal of Dermatology 16 (2006) 48\u0026ndash;55.\u003c/li\u003e\n \u003cli\u003eOnumah N., A novel anti-inflammatory in treatment of acne vulgaris: the pseudopterosins , J Drugs Dermatol 12 (2013) 1177\u0026ndash;1179.\u003c/li\u003e\n \u003cli\u003eO.M.T.S.T.Y. Amakura Y, Determination of phenolic acids in fruit juices by isocratic column liquid chromatography , J Chromatogr A 891 (2000) 183\u0026ndash;188.\u003c/li\u003e\n \u003cli\u003eT.-B.F.H.-P.B.H.D.K.A. Gil MI, Antioxidant activity of pomegranate juice and its relationship with phenolic composition and processing, J Agric Food Chem 48 (2000) 4581\u0026ndash;4589.\u003c/li\u003e\n \u003cli\u003eS.U.L.E.I.H. van der G.J. van Elswijk DA, Rapid dereplication of estrogenic compounds in pomegranate (\u003cem\u003ePunica granatum\u003c/em\u003e) using on-line biochemical detection coupled to mass spectrometry, Phytochemistry 65 (2004) 233\u0026ndash;241.\u003c/li\u003e\n \u003cli\u003eD.K.P.J.K.M.S.M.A.R.U.H.N.G. Muhammad A, Compositional analysis of natural pomegranate peel powder dried by different methods and nutritional and sensory evaluation of cookies fortified with pomegranate peel powder, Front Nutr 10 (2023).\u003c/li\u003e\n \u003cli\u003eM.J.G.W.Z.L.L.J.L.J.Z.J. Mo Y, Pomegranate peel as a source of bioactive compounds: A mini review on their physiological functions , Front Nutr 9 (2022).\u003c/li\u003e\n \u003cli\u003eS.M.A.H.K.M.A.J.N.M.A.S.I.A.K.U.Z.S.I.F.C.S.S.U.A.R.K.A.A.Y.S.H. Azmat F, Phytochemical profile, nutritional composition of pomegranate peel and peel extract as a potential source of nutraceutical: A comprehensive review, Food Science and Nutrients 12 (2024) 661\u0026ndash;674.\u003c/li\u003e\n \u003cli\u003eP.S.C.S.V.S.K.A.R.C. Dr\u0026eacute;no B, Cutibacterium acnes (Propionibacterium acnes) and acne vulgaris: a brief look at the latest updates., Journal of the European Academy of Dermatology and Venereology. 32 (2018) 5\u0026ndash;14.\u003c/li\u003e\n \u003cli\u003eA.F.K.H.N.F.N.T. Khorvash F, Staphylococcus aureus in acne pathogenesis: a case-control study. , N Am J Med Sci 4 (2012) 573.\u003c/li\u003e\n \u003cli\u003eZ.Z.S.S. Jothy SL, Phytochemicals screening, DPPH free radical scavenging and xanthine oxidase inhibitiory activities of Cassia fistula seeds extract, Journal of Medicinal Plants Research 5 (2011) 1941\u0026ndash;1947.\u003c/li\u003e\n \u003cli\u003eH.M.A.E.M.S.H.M. Alavi M, Antibacterial and antioxidant activity of catechin, gallic acid, and epigallocatechin-3-gallate: Focus on nanoformulations, Cellular, Molecular and Biomedical Reports 3 (2023) 62\u0026ndash;72.\u003c/li\u003e\n \u003cli\u003eK.H. Salehi-Sardoei A, Nitric oxide signaling pathway in medicinal plants, Cellular, Molecular and Biomedical Reports 2 (2022) 1\u0026ndash;9.\u003c/li\u003e\n \u003cli\u003eN. da S.L.M. da F.C.D.A.J.C.M.C.M.L.VL. Silva AP, Antimicrobial activity and phytochemical analysis of organic extracts from cleome spinosa Jaqc, Front Microbiol 7 (2016) 963.\u003c/li\u003e\n \u003cli\u003eM.S.P.D.H.A.C.MA. Bhattacharya SA, HPLC of phenolic compounds, antioxidant and antimicrobial activity of bulbs from three Ornithogalum species available in India. , Int J Pharm Pharm Sci 8 (2016) 187\u0026ndash;192.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-7628493/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7628493/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAcne is a widespread skin ailment characterized by the growth of pimples, blackheads, and cysts, typically on the face, neck, back, and chest. It originates from blocked hair follicles, oil (sebum) buildup, and dead skin cells, leading to bacterial development and irritation. Herbal formulations have gained appeal due to their safety and efficacy, notably in the beauty business. Pomegranate peels (\u003cem\u003ePunica granatum\u003c/em\u003e) are rich in antibacterial and antioxidant components, making them appropriate for managing acne.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eThe Main body of the abstract\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research focused on developing an herbal anti-acne gel utilizing hydro-alcoholic extracts of Pomegranate peels. Gel was formulated to increase the bioavailability and penetration of the active components. The gel's physicochemical qualities, including pH, viscosity, spreadability, and swelling index, were examined and shown to be suitable for skin application, allowing simplicity of use and effective delivery. The extract was characterized using UV-visible spectroscopy to identify key bioactive compounds. Furthermore, the existence of strong free radical scavengers was confirmed by DPPH tests, which were used to assess antioxidant activity. The anti-inflammatory potential of the gel was assessed through in vitro studies, demonstrating significant inhibition of inflammatory mediators.\u003c/p\u003e\n\u003cp\u003eThe gel formulation exhibited substantial antibacterial activity against Propionibacterium acnes and Staphylococcus aureus, surpassing the efficacy of standard Clindamycin. Stability experiments conducted over three months at regulated temperatures (25±2°C and 2-8±3°C) proved the gel's endurance and preserved its physicochemical integrity. This study emphasizes the advantages of using Pomegranate Peel Extract in gel form for acne treatment. Benefits include enhanced bioavailability, potent antioxidant and anti-inflammatory activity, and reduced discomfort. For those looking for safe and efficient acne treatments, the mixture offers a potential natural substitute.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eShort Conclusion\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe produced Pomegranate Peel Extract gel revealed remarkable anti-acne effects, stability, and skin compatibility. Its formulation is safe, easy, and efficient, giving substantial potential for application as an anti-acne ingredient in cosmetic and dermatological products.\u003c/p\u003e","manuscriptTitle":"Development and Assessment of a Pomegranate Peel-Derived Herbal Gel for Improved Anti-Acne Efficacy and Skin Tolerance","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-13 14:44:56","doi":"10.21203/rs.3.rs-7628493/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"cf23ce94-7ff6-42ba-a9f9-7b4eab3b4ed9","owner":[],"postedDate":"October 13th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-10-27T14:40:10+00:00","versionOfRecord":[],"versionCreatedAt":"2025-10-13 14:44:56","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7628493","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7628493","identity":"rs-7628493","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}