Phytochemical and Biological Evaluation of a Newly Designed Vitis Vinifera Seed Oil Self-Nanoemulsifying System

Phytochemical and Biological Evaluation of a Newly Designed Vitis Vinifera Seed Oil Self-Nanoemulsifying System | 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 Phytochemical and Biological Evaluation of a Newly Designed Vitis Vinifera Seed Oil Self-Nanoemulsifying System Ahmad M Eid, Linda Issa, Haneen safadi, Rola sabbah, Sondos mabrokeh, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4467611/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 The objective of this research is to extract and formulate a nanoemulsion consisting of Vitis vinifera ( V. vinifera ) oil. Following this, the antioxidant, anti-diabetic, and anti-lipase properties of both the oil and the nanoemulsion system will be examined. Method V. vinfera seed oil extraction was followed by the preparation of its self-nanoemulsifying system. GC analysis was conducted to analyze phytochemical compounds, and physical characterization was performed for the V. vinfera oil self-nanoemulsifying system. Subsequently, antioxidant, anti-diabetic, and anti-lipase activities were investigated for the oil and its nanoemulsion. Result The biologically active components of V. vinifera oil were detected using a GC/MS device. The main components were: linoleic acid methyl ester, oleic acid methyl ester, palmitic acid methyl ester, and stearic acid methyl ester, and elaidic acid methyl ester. The optimum nanoemulsion formulation, which comprised Tween 80, Span 80, and V. vinifera seed oil and had a PDI of 0.227 ± 0.07 and a droplet size of 192.71 ± 1.8 nm. Porcine lipase inhibition of V. vinifera oil and its nanoemulsion was detected; the oil showed potent lipase inhibitory activity when compared to orlistat. The self-nanoemulsion has less inhibitory activity than the oil, with an IC 50 equivalent to 13.8 ± 1.5 µg/ml. Also, V. vinifera oil inhibited a-amylase with a weak IC 50 value of 257.03 ± 2.4 µg/ml. The free radical scavenging activity of the extracted V. vinifera oil was estimated using the DPPH radical method and trolox. The self-nanoemulsion has less inhibitory activity than the oil, with an IC 50 value of 13.80 ± 1.5 µg/ml. Conclusion This study demonstrated the importance of black grape ( V. vinfera ) oil for treating and preventing obesity, diabetes, and oxidative stress, as well as how nanoemulsions improve these activities. Therefore, this natural oil is a promising product for the pharmaceutical industry. Vitis Vinifera self-nanoemulsifying system Phytochemical Antioxidant Anti-diabetic anti-lipase Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 1. Introduction A wide range of therapeutic plants exist in the world. Prehistoric human beings stumbled upon the healing properties of medicinal plants through trial and error in their quest to alleviate disease symptoms. The general consensus is that natural remedies are safer, more benign, and healthier than synthetic ones. Therefore, many diseases have been treated with medicinal plants [ 1 , 2 ]. In the past, natural chemicals and similar structures have been important in pharmacology, especially for treating infectious disorders and cancer. There are around 100 new products now undergoing clinical development particularly for these disorders [ 3 ]. In order to facilitate the utilization of natural products in drug discovery efforts, many screening methodologies are being devised. Data mining and virtual screening approaches are being used to databases that include information on natural products. Utilizing natural materials in a more effective and efficient manner is anticipated to improve the process of discovering new drugs [ 4 ]. The swift progress of technology has led to the emergence of innovative methods in the quest for pharmaceuticals obtained from natural sources. The utilization of artificial intelligence and bioinformatics has facilitated the examination and production of natural products [ 5 ]. Vitis vinifera ( V. vinifera ) is a perennial plant belonging to the vitaceae family. The primary bioactive components found in V. vinifera seed oil are phenolic compounds, which encompass flavonoids, tannins, phenolic acids, carotenoids, and stilbenes. Grape seed oil contains phenolic components such as catechins, epicatechins, and gallic acid, as well as fatty acids including linoleic acid, vitamin E, and phytosterols. The seeds have an elliptical or pyriform morphology with a tapered apex. Their length can extend up to 6 mm [ 6 , 7 ]. V. vinifera seed possesses antioxidant, antifungal, antidiabetic, anti-inflammatory, and anti-obesity properties, as well as exhibiting anticholinergic scarring effects. V. vinifera seed oil includes essential components such as ascorbic acid, beta-carotene, phytosterols, and tocopherols. These chemicals, which are present in the stems and leaves of the plant, provide protection against oxidative stress. Goufo, Singh, and Cortez (2020) found that the presence of proanthocyanidins in the seeds is indicative of the oil's antioxidant potential. Scientific study has demonstrated that V. vinifera seed possess distinct biological and therapeutic characteristics, making them advantageous for one's health. However, a limited number of research that employ experimental models of obesity, oxidative stress, or inflammation document the therapeutic capacity of these models in treating metabolic diseases [ 8 ]. Research has shown that V. vinifera seed oil effectively inhibits the rapid development of human cancer cells. Grape seed oil contains organic fatty acids, including malic, tartaric, and oxalic acids, which have the ability to reduce LDL-c levels [ 9 ]. V. vinifera seed oil is rich in tocotrienols, particularly α- and γ-tocotrienol (T3), which effectively decrease the production of mRNA protein. This reduction is crucial for inhibiting the process of adipogenesis, namely the activity of PPARγ and aP2. Furthermore, it substantially reduces the production of pro-inflammatory genes, including IL-6 and IL-8 [ 10 ]. The presence of polyphenols, including flavanols, catechin, and epicatechin, in this function further improves its effectiveness. These polyphenols may be associated with preventing arterial thrombosis. Additionally, compounds such as flavones, isoflavones, and anthocyanins are involved in preventing pathologies related to weight gain and possess anti-inflammatory properties. [ 7 , 11 ]. V. vinifera has been used as a key component in the cosmetic sector due to its significant antioxidant, antibacterial, and skin care attributes [ 12 ]. he CosIng (cosmetic ingredients) database states that cosmetics utilize grape-derived compounds in nine different ways [ 13 ]. The extract of V. vinifera can serve as an emollient, humectant, emulsifier, color addition, or fragrance. For instance, grape seeds are suggested for use as a color, moisturizer, or hair and skin care product. Grape-derived cosmetics possess gentle and non-aggravating properties when applied to the skin. Scientific study indicates that this plant possesses antioxidant, whitening, anti-inflammatory, anti-aging, and other properties mostly because of its abundant phenolic and stilbene constituents. It is worth noting that V. vinifera oil or extracts has distinctive moisturizing and anti-aging properties, as evidenced by the increasing use of these fundamental constituents in cosmetic goods [ 14 – 16 ]. According to Wijekoon et al., ingesting grape varietals with seeds is important as a functional diet since they contain considerable amounts of anthocyanins, flavones, flavonols, and stilbenes (specifically resveratrol) [ 17 ]. Argon et al. state that grape seed oil includes bioactive chemicals such as carotenoids, tocopherols (tocols), phytosterols, phenolic components, and other fat-soluble compounds. Tocopherols are powerful, naturally-occurring antioxidants that dissolve in fats [ 18 ]. An experiment conducted on mice revealed that those mice who were fed a hyperlipidic diet enriched with 15.5 g of grape seed oil per 100 g of food for a duration of eight weeks had reduced levels of glucose and total cholesterol in their blood plasma, as well as enhanced glucose tolerance. Furthermore, the subjects exhibited reduced weight gain and decreased weight of white adipose tissue. Additionally, they demonstrated significant antioxidant properties and lower levels of TBARS in their plasma, which are indicators of lipid peroxidation. Moreover, there was a decrease in the production of IL-6 and IL-10. Based on the evidence, it can be inferred that V. vinifera seed oil is classified as a functional oil that has the capacity to decrease inflammation and obesity issues caused by oxidative stress [ 19 ]. Nanotechnology refers to the manipulation and creation of materials at extremely small sizes, namely at the atomic and molecular levels [ 20 ]. Scientists are developing novel formulations such as nanoemulsions [ 21 ] to address the limitations of existing formulations, which include inadequate targeting of particular sites, poor spreading ability, and insufficient bioavailability. The nanoemulsion (NE) system is characterized by its stability, viscous formulation, tiny particle size, high encapsulation efficiency, good thermodynamic stability, and strong penetration ability [ 22 ]. Nanoemulsions consist of droplets with a size range of 10 to 100 nm, allowing them to quickly enter and distribute active ingredients profoundly [ 23 , 24 ]. Self-emulsifying drug delivery methods are crucial for resolving the issue of low bioavailability associated with drugs that have poor solubility [ 25 ]. Repeatedly, the use of nutraceuticals has been linked to safeguarding against chronic ailments such as diabetes, heart disease, cancer, and neurological disorders. However, the therapeutic potential of these nutraceuticals is limited due to their low permeability and poor solubility in water. Therefore, the utilization of self-emulsifying formulation has demonstrated promising potential in improving the absorption rate and effectiveness of nutraceuticals, addressing the aforementioned challenges [ 26 ]. This work aims to extract, analyze the phytochemical characteristics, and assess the biological activities of V. vinifera seed oil and its self-nanoemulsifying formulation. 2. Materials and Methods 2.1. Materials Dimethyl sulfoxide (DMSO) was obtained from Riedel De Haen, Germany, and p-nitrophenyl butyrate (PNPB) from Sigma-Aldrich, Germany. Sigma-Aldrich (USA) provided acarbose, porcine pancreatic lipase type 2, starch, orlistat, (p-nitrophenyl α-D-galactopyranoside) PNPG, and 2,2-diphenyl-1-picrylhydrazyl (DPPH). Both Span 80 and Tween 80 were obtained from the Al-Shams company (Palestine). Trolox [(S)-(-)-6-hydroxy-2, 5, 7, 8-tetramethylchroman-2-carboxyylic acid]. The α-amylase enzyme from Sigma-Aldrich (India) and n-hexane (Frutarom Ltd., Israel). 2.2. Extraction of the V. vinifera seed essential oil The V. vinifera fruits were collected in 2023 from Hebron in Palestine. The fruits were then identified at the pharmacognosy laboratory at An-Najah National University using the voucher specimen code Pharm-PCT-2665. The fruit seeds were dried in the shade at a controlled temperature [25 ± 2°C] and humidity [55 ± 4 RH]. Following the drying process, we ground the seeds into a fine powder. About 50 g of the powdered V. vinifera seeds were soaked in 250 mL of 70% ethanol, a polar solvent, and 250 mL of hexane, a non-polar organic solvent, for 72 hours in a shaker at room temperature. Afterwards, we divide the mixture into two layers using a separotary funnel: the upper layer, the organic phase containing the oil, and the lower layer, the aqueous phase. Then the solvent was removed under air pressure to concentrate the organic layer [ 27 – 29 ]. 2.3. Bioactive components study of V. vinifera seed oil by GC-MS Using the gas chromatography-mass spectrometry (GC-MS) method, the fatty acid methyl ester (FAME) of V. vinifera seed oil was analyzed. To prepare the oil, we mixed 100 mg of V. vinifera seed oil with 10 ml of a 0.5 M methanolic sodium hydroxide solution, then heated the mixture for complete hydrolysis on a heating mantel for 10 minutes. After that, I added 10 ml of 14% boron trifluoride (BF3) solution in methanol and refluxed them for 10 minutes. After the esterification, add 50 ml of heptan and continue the boiling for 10 minutes, then the mixture was cooled. A Perkin Elmer Clarus 500 gas chromatograph and a Perkin Elmer Clarus 560 mass spectrometer were used to examine the FAME of V. vinifera seed oil in the heptan layer. A fused silica capillary column (30 m x 0.25 mm, film thickness 0.25 µm) was used. The column temperature was planned to increase by 4°C/min, so the temperature of the well increased from 3°C to 240°C (hold for 15 minutes). Helium gas (99.999%) is used as a carrier with a flow rate of 1 ml/min. With a split ratio of 50:1 and at a temperature of 25°C, 1 µl of oil was injected in split mode. Solvent delay: 0–8 min. The entirety of the data was obtained through the collection of full-scan mass spectra spanning the range of 50–500 atomic mass units (amu). Ionizing energy measured 70 eV. The voltage of the electron multiplier (EM) was acquired via autotune. The mass spectrum of the chemical components of V. vinifera oil was compared to the reference spectrum from the MS Database of the National Institute of Standards and Technology, and their retention and kovats results were compared to other research results [ 27 – 29 ]. 2.4. Preperation of V. vinifera seed oil nanoemulsion Using the self-nanoemulsifying technique, we convert the extracted V. vinifera seed oil into a nanoemulsion following extraction. To construct the ternary phase diagram, three different substances were used at varying concentrations: V. vinifera seed oil, Span 80 as a co-surfactant, and Tween 80 as a surfactant! After determining the weight of each formulation using a weighting balance, the mixture was agitated gently for three minutes with a vortex mixer in order to achieve homogeneity. Then, each formulation was emulsified under gentle agitation with distilled water. The formulation that yields the highest possible particle size and polydispersity index (PDI) has been determined [ 30 , 31 ]. 2.5. Droplet size and PDI analysis of V. vinifera seed oil nanoemulsion To determine the droplet size and PDI, a master size analyzer (Brookhaven Instruments, NanoBrook Omni, New York) was used. The V. vinifera seed oil nanoemulsion was self-emulsified in distilled water before using the measuring procedure. The ideal nanoemulsion formulation was chosen based on the smallest droplet size and PDI and with the highest amount of V. vinifera seed oil [ 32 ]. 2.6. Antioxidant activity of V. vinifera seed oil Initially, methanol was used to produce stock solutions (1 mg/ml) of V. vinifera seed oil and its self-nanoemulsifying formulation. At the following concentrations (2, 5, 10, 20, 50, 80, and 100 µg/ml), each of the stock solutions was diluted in methanol. A DPPH solution that had been recently prepared was dissolved in methanol at a concentration of 0.002% w/v. Following that, a 1:1:1 ratio was maintained between each working solution and methanol and DPPH, with the lowest concentration added last. The solutions were incubated in a dark area at room temperature for thirty minutes. As a baseline, methanol was utilized on the UV-visible spectrophotometer; all absorbance values were calculated at a wavelength of 517 mm. Trolox was employed in the capacity of a positive control. Determine the percentage of antioxidant activity exhibited by each tested compound and the Trolox standard by employing the subsequent equation: [ 33 ]. % DPPH inhibition = (A b – A s ) / A b × 100% A b : blank absorbance, and A s : sample absorbance. 2.7. Anti-diabetic activity of V. vinifera seed oil using α-amylase inhibitory method The inhibitory effect of the α-amylase enzyme by V. vinifera seed oil and its self-nanoemulsifying formulation was evaluated using a modified version of a standard technique that had been previously established [ 34 ]. In order to achieve a concentration of 1000 µg/mL, the oil and its self-nanoemulsion in 10% DMSOO were dissolved individually in a buffer solution consisting of NaCl and Na2HPO4. Several dilutions containing 10, 50, 70, 100, and 500 µg/mL were subsequently performed. After combining 0.2 mL of the oil with an equivalent volume of panreatic amylase enzyme solution, the mixture was incubated at 30°C for a duration of 10 minutes. The mixture was then incubated for three minutes after 0.2 mL of starch solution was added. 0.2 mL of DNSA is introduced to halt the incubation process. The mixture was then diluted with 5 mL of distilled water and heated for 10 minutes at 90°C in a water bath. 540nm absorption measurements were taken of the mixtures subsequent to their reduction to room temperature. To produce the blank, a buffer solution was substituted for the oil. For the positive control, which is a carbohydrate, the exact same procedure steps were followed. Using the equation below, The inhibitory activity of α-amylase was assessed [ 35 ]. % α- amylase inhibition = (A b – A s ) / A b × 100% A b : blank absorbance, and A s : tested sample absorbance. 2.8. Anti-obesity acivity of V. vinifera seed oil using lipase inhibitory method Stock solutions of orlistat, a positive control, and V. vinifera seed oil and its self-nanoemulsion were prepared at a concentration of 1000 µg/mL. Subsequently, the solutions mentioned above are diluted in a test tube containing 50, 100, 200, 300, and 400 µg/mL. A bulk solution of pancreatic lipase enzyme was formulated in a 10% DMSO solution. Following that, 0.1 mL of pancreatic lipase enzyme and 0.7 mL of buffer containing Tris-HCl were added to the solution. Each test tube, along with a blank tube containing Tris-HCl and pancreatic lipase enzyme, was incubated at 37°C for a duration of 15 minutes. Additional incubation for 30 minutes in the blank and each test tube subsequent to adding 0.1 mL of PNPB. In order to evaluate the hydrolysis of PNPB to p-nitophenol, a 410 nm UV-visible spectrometer was employed [ 36 ]. % lipase inhibition = (A b – A s ) / A b × 100% A b : blank absorbance, and A s : sample absorbance. 3. Results 3.1. Analysis of V. vinifera seed oil chemical components using GC-MS The biologically active components of V. vinifera seed oil were detected using a GC-MS device, as shown in Fig. 1 . The main components showed in Table 1 were linoleic acid methyl ester (62.97%), oleic acid methyl ester (23.07%), palmitic acid methyl ester (8.36%), stearic acid methyl ester (4.96%), and elaidic acid methyl ester (0.64%). Table 1 Natural active compounds characterized by GC-MS of V. vinifera seed oil Fatty acid of grape seed oil % age Palmitic acid 8.36% Linoleic acid 62.97% Oleic Acid 23.07% Elaidic Acid 0.64% Stearic Acid 4.96% 100% 3.2. Droplet size and PDI analysis of Vitis Vinifera nanoemulsion formulations In order to determine the most effective formulation, ternary phase diagrams were constructed by employing different concentrations of Span 80, Tween 80, and V. vinifera seed oil (Fig. 2 ). Subsequently, a nanoemulsion was generated characterized by a droplet size below 200 nm and a PDI below 0.2. In order to conduct a comparative analysis of the chosen formulations exhibiting droplet sizes below 200 nm, we employed the optimal nanoemulsion formulation consisting of 35% V. vinifera seed oil, 50% Tween 80, and 15% Span 80 (Table 2 ). This formulation exhibited a PDI of 0.227 ± 0.07 and a droplet size of 192.71 ± 1.8 nm. Table 2 The selected optimum Vitis Vinifera seed oil self-nanoemulsifying formulation. Tween 80(%) Span 80 (%) Oil(%) Droplet size(%) Polydispersity Index(PDI) 50 15 35 192.71 ± 1.8 0.227 ± 0.07 3.3. Antioxidant activity of V. vinifera seed oil and its self-nanoemulsifying formulation The DPPH radical method was employed to determine the free radical scavenging activity of the extracted V. vinifera seed oil and it self-nanoemulsifying formulation, with trolox serving as positive control. The prospective antioxidant activity of the essential oil of V. vinifera was demonstrated in Fig. 3 , where it exhibited an IC 50 value of 33.88 ± 1.7 µg/ml, in contrast to the positive control (Trolox) which showed an IC 50 value of 4.28 ± 0.6 µg/ml. The oil possesses a lower inhibitory activity than the self-nanoemulsion, as evidenced by its IC 50 value of 13.80 ± 1.5 µg/ml. 3.4. Anti-diabetic activity of V. vinifera seed oil and its self-nanoemulsifying formulation Figure 4 shows the percentage of α-amylase inhibition for V. vinifera seed oil and its self-nanoemulsifying formulation at various concentrations compared to acarbose as a positive control. The IC 50 values for V. vinifera seed oil and its self-nanoemulsifying formulation were 257.03 ± 2.4 µg/ml and 52.48 ± 1.2 µg/ml, respectively. These values were not as strong as the IC 50 value for acarbose, which was 39.81 ± 0.8 µg/ml. The oil activity improved when it was converted to a self-nanoemulsifying system and showed an IC 50 close to that of the positive control. 3.5. Anti-obesity activity of V. vinifera seed oil and its selfnanoemulsifying formulation V. vinifera seed oil and its self-nanoemulsifying formulation exhibited porcine lipase inhibition; this was subsequently evaluated in comparison to orlistat, which serves as the positive control. The lipase inhibitory activity of the oil is significantly high when compared to that of orlistat, as illustrated in Fig. 4 . IC 50 values for the oil and orlistat were 33.88 ± 1.7 µg/ml and 12.3 ± 0.8 µg/ml, respectively. With an IC 50 value of 13.8 ± 1.5 µg/ml, the self-nanoemulsifying formulation of V. vinifera seed oil exhibits superior inhibitory activity in comparison to the oil, specifically orlistat. 4. Discussion The chemical constituents of V. vinifera seed oil were identified using a Gas Chromatography-Mass Spectrometry (GC-MS) instrument, as seen in Fig. 1 . The primary constituents were of linoleic acid methyl ester (62.97%), oleic acid methyl ester (23.07%), palmitic acid methyl ester (8.36%), stearic acid methyl ester (4.96%), and elaidic acid methyl ester (0.64%). The study conducted by Al-Fekaiki and Al-Hilfi. The fatty acid composition of V. vinifera seed oil was determined in 2016 using GC-MS. The analysis revealed the presence of various fatty acids, including oleic acid (16.98%), palmitic acid (12.09%), stearic acid (8.85%), palmitoleic acid (0.28%), myristic acid (0.21%), azelaaldehydic acid (0.12%), margaric acid (0.11%), lauric acid (0.05%), and pentadecanoic acid (0.03%) [ 37 ]. A recent study conducted by Sabir, Unver, and Kara investigated the composition of V. vinifera seed oil and found varying percentages of different fatty acids. The major component was linoleic acid, accounting for 53.6–69.6% of the oil. This was followed by oleic acid (16.2–31.2%), palmitic acid (6.9–12.9%), and stearic acid (1.44–4.69%). Additionally, we discovered many chemical constituents such as alpha-tocopherol, beta-tocopherol, gamma-tocopherol, and sigma-tocopherol. The oil samples from grape seeds include much larger quantities of linoleic acid and substantial levels of tocopherols, indicating that grape seed is a valuable source for culinary, medicinal, and cosmetic applications [ 38 ]. A further investigation on V. vinifera seed oil revealed a substantial quantity of phenolic substances, encompassing flavonoids, carotenoids, phenolic acid, tannins, and stilbenes. Gallic acid possesses antioxidant properties and contains polyphenols such as catechins, epicatechins, trans-resveratrol, and procyanidin B1 [ 7 ]. By creating formulations with varying concentrations of V. vinifera seed oil, surfactant, and co-surfactant, we constructed a ternary phase diagram to identify the optimal formulation for the self-nanoemulsifying drug delivery system (SNEDDS). The goal was to achieve a droplet size smaller than 200 nm and a PDI (polydispersity index) lower than 0.2. Tween 80 and Span 80 are non-ionic surfactants that make up 50% and 15% of our composition, respectively. Multiple studies have demonstrated a direct correlation between an increase in surfactant content and a decrease in droplet size. Greater quantities of surfactant are needed to stabilize smaller droplets since they have a larger surface area [ 39 ]. Moreover, a higher quantity of surfactant is necessary to stabilize diminutive particles that possess a larger surface area. Recent study suggests that an elevation in the surfactant concentration in our formulation might result in several outcomes, such as a reduction in surface tension and an expansion in artificial area, possibly due to an enlargement in particle size [ 40 ]. Singh et al. (2017) shown that nanoemulsion systems are highly effective in enhancing medication bioavailability, delivery, solubility, stability, and effectiveness [ 41 ]. A self-nanoemulsifying system was created to boost the bioactivity of V. vinifera seed oil, resulting in greater solubility and hence higher efficacy [ 42 ]. Wang et al. (2023) conducted a research to examine the advantageous characteristics of lingonberry leaves and their active constituents. The study revealed that these features were further increased when included into nanoemulsions. The study's findings were acquired concurrently using internet means. The purpose of developing nanoemulsions was to enhance the stability and antioxidant effectiveness of extracts from lingonberry leaves. The nanoemulsions exhibited enhanced antioxidant activity, with a DPPH radical inhibition rate exceeding 13% at the same dose. This enhancement was statistically significant when compared to their non-nanoemulsified counterparts. The study discovered that nanoscale emulsions greatly enhance the solubility of polyphenols, leading to the optimization of their antioxidant capabilities [ 43 ]. Multiple studies have shown that grape seeds possess strong antioxidant properties, possibly attributed to the presence of different phenolic compounds such as epicatechin, catechin, procyanidins, proanthocyanidins, gallic acid, and ellagic acid [ 44 ]. Several investigations have demonstrated that V. vinifera seed oil exhibits other actions beyond its antioxidant properties. These include anti-inflammatory, anti-cancer, anti-microbial, anti-ulcer, and cardioprotective benefits [ 45 ]. The research conducted by Pérez et al. (2015) confirmed that grape seed oil extract had remarkable antioxidant properties, with a scavenging activity of 95.8% against DPPH radicals [ 46 ]. Our results were consistent with this finding, as the V. vinifera seed oil exhibited noteworthy antioxidant properties. The IC 50 value for the V. vinifera seed oil was 33.88 ± 1.7 µg/ml, while the self-nanoemulsifying system had an IC 50 value of 13.80 ± 1.5 µg/ml, indicating even stronger antioxidant activity. In comparison to trolox, which had an IC 50 value of 4.28 ± 0.6 µg/ml, the self-nanoemulsifying system demonstrated superior antioxidant activity compared to the pure oil. Obesity and diabetes are becoming prevalent health problems worldwide. They have been recognized as major risk factors for many infections, complications following infections, and death caused by severe infections [ 47 ]. The study conducted by Ahmed et al. investigated the antidiabetic effects of grape stem extract. The results showed that both the chloroform and ethanolic stem extracts were able to lower blood glucose levels in a dose-dependent way. The maximum activity was shown at a dosage of 200 mg/kg bodyweight [ 48 ]. The IC 50 value for V. vinifera seed oil was determined to be 257.03 ± 2.4 µg/ml, while the IC 50 value for the oil self-nanoemulsifying system was found to be 33.88 ± 1.6 µg/ml. These values, although relatively moderate, were compared to the reference compound Acarbose, a potent α-amylase inhibitory agent, which exhibited an IC 50 value of 39.81 ± 0.8 µg/ml. The aqueous extracts of V. vinifera exhibited the most potent anti-lipase activities, as indicated by their IC 50 values of 14.13µg/ml. V. vinifera exhibited inhibitory effects on pig pancreatic lipase compared to Orlistat, with an IC 50 value of 12.38 µg/ml. This work was performed on V. vinifera and its self-nanoemulsifying system. The aqueous extracts of V. vinifera exhibited strong anti-lipase actions, with IC 50 values of 14.13 µg/ml. Concurrently, V. vinifera exhibited inhibitory effects on pig pancreatic lipase compared to orlistat, with an IC 50 value of 12.38 µg/ml [ 49 ]. A nanoemulsion was created to enhance the bioactivity of V. vinifera seed oil. The improvement occurred because the nanoemulsion had a higher solubility compared to the oil and other drugs. This is because the nanoemulsion has a larger surface area and very small particle size, which both contribute to its increased effectiveness [ 50 ]. 5. Conclusion The present study investigated the anti-diabetic, antioxidant, and anti-obesity effects of V. vinifera nanoemulsion in comparison to pure oil and positive control drugs. The undiluted oil exhibited anti-obesity effects against lipase, antioxidant effects against DPPH, and anti-diabetic effects against α-amylase; these effects were enhanced when the oil was in the form of a nanoemulsion. In comparison to the unadulterated V. vinifera seed oil, the nanoemulsion exhibited noteworthy antioxidant, anti-diabetic, and anti-obesity properties, a reduction in droplet size, and an enhancement in stability and solubility. The results indicate that V. vinifera seed oil is a valuable reservoir of bioactive and antioxidant substances, particularly unsaturated fatty acids such as linoleic acid, palmitic acid, oleic acid, and stearic acid. This study proposes that incorporating V. vinifera seed oil into nanoemulsions can be a highly effective method for formulating oral medications. Hence, owing to its capacity to hinder the activity of α-amylase, lipase, and free radicals in comparison to V. vinifera seed oil and Trolox, it has the potential to be employed as a supplementary treatment for obesity and diabetes, alongside other drugs, in order to impede the advancement of these disorders. Furthermore, as demonstrated by this work, a self-nanoemulsion system may serve as a reservoir for drugs, improve the way drugs are absorbed and distributed in the body, and facilitate the release of oil in oil-based therapeutic formulations. Declarations Permissions and identification of plant material Permissions were obtained to collect the plant material. The plant material was collected and identified within the Pharmacy Department of An-Najah National University. Ethical approval Authors confirm that the used of plant in the present study, including the collection of plant material, comply with relevant institutional, national, and international guidelines and legislation. Data availability The manuscript file contains all of the data that was created or analyzed for this project. I confirm that the study has all the information and consents required. Competing interests The authors declare that they have no competing interests. Funding None. Authors' contributions AE: conceptualization, formal analysis, supervision, writing—original draft. LI, HS, RS, SM, and TH carried out the formal analysis, Acknowledgments The authors would like to acknowledge the Faculty of Medicine and Health Sciences at An-Najah National University. References Mannangatti P, Naidu KN. Indian Herbs for the Treatment of Neurodegenerative Disease. Adv Neurobiol. 2016;12:323–36. Gozubuyuk GS, Aktas E, Yigit N. An ancient plant Lawsonia inermis (henna): determination of in vitro antifungal activity against dermatophytes species. J Mycol Med. 2014;24(4):313–8. Harvey AL. Natural products in drug discovery. Drug Discov Today. 2008;13(19–20):894–901. 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Comparative Protective Effect of Nigella sativa Oil and Vitis vinifera Seed Oil in an Experimental Model of Isoproterenol-Induced Acute Myocardial Ischemia in Rats. Molecules. 2021;26(11). Zhao L, Yagiz Y, Xu C, Lu J, Chung S, Marshall MR. Muscadine grape seed oil as a novel source of tocotrienols to reduce adipogenesis and adipocyte inflammation. Food Funct. 2015;6(7):2293–302. Gomes JMG, Costa JA, Alfenas RCG. Metabolic endotoxemia and diabetes mellitus: A systematic review. Metabolism. 2017;68:133–44. Fiume MM, Bergfeld WF, Belsito DV, Hill RA, Klaassen CD, Liebler DC, et al. Safety assessment of Vitis vinifera (grape)-derived ingredients as used in cosmetics. Int J Toxicol. 2014 Sep-Oct;33(3 Suppl):s48–83. Di Pietro Fernandes C, Santana LF, Dos Santos JR, Fernandes DS, Hiane PA, Pott A et al. Nutraceutical Potential of Grape (Vitis vinifera L.) Seed Oil in Oxidative Stress, Inflammation, Obesity and Metabolic Alterations. Molecules. 2023;28(23). Sharafan M, Malinowska MA, Ekiert H, Kwaśniak B, Sikora E, Szopa A. Vitis vinifera (Vine Grape) as a Valuable Cosmetic Raw Material. Pharmaceutics. 2023;15(5). Lin YS, Chen HJ, Huang JP, Lee PC, Tsai CR, Hsu TF, et al. Kinetics of Tyrosinase Inhibitory Activity Using Vitis vinifera Leaf Extracts. Biomed Res Int. 2017;2017:5232680. Di Lorenzo C, Sangiovanni E, Fumagalli M, Colombo E, Frigerio G, Colombo F et al. Evaluation of the Anti-Inflammatory Activity of Raisins (Vitis vinifera L.) in Human Gastric Epithelial Cells: A Comparative Study. Int J Mol Sci. 2016;17(7). Wijekoon C, Netticadan T, Siow YL, Sabra A, Yu L, Raj P et al. Potential Associations among Bioactive Molecules, Antioxidant Activity and Resveratrol Production in Vitis vinifera Fruits of North America. Molecules. 2022;27(2). Ustun Argon Z, Celenk VU, Gumus ZP. In: Ramadan MF, editor. Chapter 5 - Cold pressed grape (Vitis vinifera) seed oil. Cold Pressed Oils: Academic; 2020. pp. 39–52. Martínez-Galán JP, Ontibón-Echeverri CM, Campos Costa M, Batista-Duharte A, Guerso Batista V, Mesa V, et al. Enzymatic synthesis of capric acid-rich structured lipids and their effects on mice with high-fat diet-induced obesity. Food Res Int. 2021;148:110602. Farokhzad OC, Langer R. Impact of nanotechnology on drug delivery. ACS Nano. 2009;3(1):16–20. Rathee J, Malhotra S, Pandey M, Jain N, Kaul S, Gupta G et al. Recent Update on Nanoemulsion Impregnated Hydrogel: a Gleam into the Revolutionary Strategy for Diffusion-Controlled Delivery of Therapeutics. AAPS PharmSciTech. 2023 2023/07/12;24(6):151. Ma Q, Zhang J, Lu B, Lin H, Sarkar R, Wu T et al. Nanoemulgel for Improved Topical Delivery of Desonide: Formulation Design and Characterization. AAPS PharmSciTech. 2021 2021/05/24;22(5):163. Gupta SK, editor. Formulation and Evaluation of Nanoemulsion Based Nanoemulgel of Aceclofenac2020. Ojha B, Jain VK, Gupta S, Talegaonkar S, Jain K. Nanoemulgel: a promising novel formulation for treatment of skin ailments. Polym Bull. 2022;79(7):4441–65. 2022/07/01. Kohli K, Chopra S, Dhar D, Arora S, Khar RK. Self-emulsifying drug delivery systems: an approach to enhance oral bioavailability. Drug Discov Today. 2010;15(21–22):958–65. Dhritlahre RK, Ruchika, Padwad Y, Saneja A. Self-emulsifying formulations to augment therapeutic efficacy of nutraceuticals: From concepts to clinic. Trends Food Sci Technol. 2021 2021/09/01/;115:347 – 65. Jaradat N, Al-Lahham S, Abualhasan MN, Bakri A, Zaide H, Hammad J, et al. Chemical Constituents, Antioxidant, Cyclooxygenase Inhibitor, and Cytotoxic Activities of Teucrium pruinosum Boiss. Essential Oil. Biomed Res Int. 2018;2018:4034689. Vinaixa M, Schymanski EL, Neumann S, Navarro M, Salek RM, Yanes O. Mass spectral databases for LC/MS- and GC/MS-based metabolomics: State of the field and future prospects. TRAC Trends Anal Chem. 2016;2016(04/01/):78:23–35. Wei X, Koo I, Kim S, Zhang X. Compound identification in GC-MS by simultaneously evaluating the mass spectrum and retention index. Analyst. 2014;139(10):2507–14. Eid AM, Hawash M. Biological evaluation of Safrole oil and Safrole oil Nanoemulgel as antioxidant, antidiabetic, antibacterial, antifungal and anticancer. BMC Complement Med Ther. 2021;21(1):159. Ostertag F, Weiss J, McClements DJ. Low-energy formation of edible nanoemulsions: factors influencing droplet size produced by emulsion phase inversion. J Colloid Interface Sci. 2012;388(1):95–102. Qushawy M, Mortagi Y, Alshaman R, Mokhtar HI, Hisham FA, Alattar A et al. Formulation and Characterization of O/W Nanoemulsions of Hemp Seed Oil for Protection from Steatohepatitis: Analysis of Hepatic Free Fatty Acids and Oxidation Markers. Pharmaceuticals (Basel). 2022;15(7). Jaradat N, Abualhasan M. Comparison of Phytoconstituents, Total Phenol Contents and Free Radical Scavenging Capacities between Four Arum Species from Jerusalem and Bethlehem. Pharm Sci 2016 2016/6/30;22(2):120–5. Dalli M, Daoudi NE, Abrigach F, Azizi SE, Bnouham M, Kim B, et al. In vitro α-amylase and hemoglobin glycation inhibitory potential of Nigella sativa essential oil, and molecular docking studies of its principal components. Front Pharmacol. 2022;13:1036129. Hawash M, Jaradat N, Elaraj J, Hamdan A, Lebdeh SA, Halawa T. Evaluation of the hypoglycemic effect of seven wild folkloric edible plants from Palestine. J Complement Integr Med. 2019;17(1). Jaradat N, Khasati A, Hawi M, Hawash M, Shekfeh S, Qneibi M, et al. Antidiabetic, antioxidant, and anti-obesity effects of phenylthio-ethyl benzoate derivatives, and molecular docking study regarding α-amylase enzyme. Sci Rep. 2022;12(1):3108. Al-fekaiki D, Ali S. Fatty Acids Composition by (GC-MS) and Most Important Physical Chemicals Parameters of Seed Oil Pomegranate and Grape Seeds2016. Sabir A, Unver A, Kara Z. The fatty acid and tocopherol constituents of the seed oil extracted from 21 grape varieties (Vitis spp). J Sci Food Agric. 2012;92(9):1982–7. Politova NI, Tcholakova S, Tsibranska S, Denkov ND, Muelheims K. Coalescence stability of water-in-oil drops: Effects of drop size and surfactant concentration. Colloids Surf A. 2017 2017/10/20/;531:32 – 9. Bouchemal K, Briançon S, Perrier E, Fessi H. Nano-emulsion formulation using spontaneous emulsification: solvent, oil and surfactant optimisation. Int J Pharm. 2004;280(1–2):241–51. Singh Y, Meher JG, Raval K, Khan FA, Chaurasia M, Jain NK, et al. Nanoemulsion: Concepts, development and applications in drug delivery. J Control Release. 2017;252:28–49. Zhang Y, Shang Z, Gao C, Du M, Xu S, Song H, et al. Nanoemulsion for Solubilization, Stabilization, and In Vitro Release of Pterostilbene for Oral Delivery. AAPS PharmSciTech. 2014;15(4):1000–8. 2014/08/01. Wang S, Cheng Y, Wang J, Ding M, Fan Z, Antioxidant Activity. Formulation, Optimization and Characterization of an Oil-in-Water Nanoemulsion Loaded with Lingonberry (Vaccinium vitis-idaea L.) Leaves Polyphenol Extract. Foods. 2023;12(23). Yilmaz Y, Toledo RT. Major flavonoids in grape seeds and skins: antioxidant capacity of catechin, epicatechin, and gallic acid. J Agric Food Chem. 2004;52(2):255–60. Martin ME, Grao-Cruces E, Millan-Linares MC, Montserrat-de la Paz S. Grape (Vitis vinifera L.) Seed Oil: A Functional Food from the Winemaking Industry. Foods. 2020;9(10). Pérez C, Ruiz del Castillo ML, Gil C, Blanch GP, Flores G. Supercritical fluid extraction of grape seeds: extract chemical composition, antioxidant activity and inhibition of nitrite production in LPS-stimulated Raw 264.7 cells. Food Funct. 2015;6(8):2607–13. Zhou Y, Chi J, Lv W, Wang Y. Obesity and diabetes as high-risk factors for severe coronavirus disease 2019 (Covid-19). Diabetes Metab Res Rev. 2021;37(2):e3377. Ahmed M, Chavan A, Lakshmikantha RY, Satwadi PR, Thimmappanahalli KB. Evaluation of antidiabetic activity of Vitis vinifera stem bark. J Pharm Res. 2012;5:5239–52. Jaradat N, Zaid AN, Hussein F, Zaqzouq M, Aljammal H, Ayesh O. Anti-Lipase Potential of the Organic and Aqueous Extracts of Ten Traditional Edible and Medicinal Plants in Palestine; a Comparison Study with Orlistat. Medicines. 2017;4(4):89. Wang S, Cheng Y, Wang J, Ding M, Fan Z, Antioxidant Activity. Formulation, Optimization and Characterization of an Oil-in-Water Nanoemulsion Loaded with Lingonberry (Vaccinium vitis-idaea L.) Leaves Polyphenol Extract. Foods. 2023;12(23):4256. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4467611","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":316338879,"identity":"a305d12d-1821-4d0a-add9-027116ae79f4","order_by":0,"name":"Ahmad M Eid","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA3ElEQVRIiWNgGAWjYDACCSDmYZBg7AfSBowNQPIAsVpmNpCohYFxA0glUVr4Z3cnfnhTYSG7+Xbzg4KfOxjk+G4kELDkztnNknPOSBhvu3PMwLD3DIOxJCEtDDdyN0jztkkkbruRYGDA28aQuIGQFvkbuZt/8/6TSNw8I/2D4d82hnqCWgxu5G6T5m2QSNwgkWNgDLQlwYCQFkOgFss5xySMZ9zIKTCWbZMwnHnmAX4tckCH3XhTUyfbPyN9m+HbNht5vuMEbEEGbAaQaCIBMBNw0SgYBaNgFIxUAACIU0yZ8Wy/pwAAAABJRU5ErkJggg==","orcid":"","institution":"An-Najah National University","correspondingAuthor":true,"prefix":"","firstName":"Ahmad","middleName":"M","lastName":"Eid","suffix":""},{"id":316338880,"identity":"15eeb29b-f00c-485b-9f9f-2bea553f5742","order_by":1,"name":"Linda Issa","email":"","orcid":"","institution":"An-Najah National University","correspondingAuthor":false,"prefix":"","firstName":"Linda","middleName":"","lastName":"Issa","suffix":""},{"id":316338881,"identity":"0544c29d-df17-4244-9c09-8c055fc0053e","order_by":2,"name":"Haneen 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University","correspondingAuthor":false,"prefix":"","firstName":"Tasneem","middleName":"","lastName":"Hassoun","suffix":""}],"badges":[],"createdAt":"2024-05-23 14:44:29","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4467611/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4467611/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":59380515,"identity":"8e586ce9-224a-4bc2-83ef-a42a515669ad","added_by":"auto","created_at":"2024-07-01 05:26:32","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":360964,"visible":true,"origin":"","legend":"\u003cp\u003eGas chromatography-mass spectrometry chromatogram of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4467611/v1/6328ff59999a58f5c2d07970.png"},{"id":59380517,"identity":"e5cf0e15-cc90-496d-8502-2e6178523e06","added_by":"auto","created_at":"2024-07-01 05:26:32","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":138241,"visible":true,"origin":"","legend":"\u003cp\u003ePseudo ternary phase diagrams of\u003cem\u003e Vitis Vinifera\u003c/em\u003e\u003cstrong\u003e \u003c/strong\u003eseed oil self-nanoemulsifying formulations.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4467611/v1/748d54ebeb471df87ab2fde1.png"},{"id":59380516,"identity":"2befc39e-6b32-4001-aee8-689addbb279b","added_by":"auto","created_at":"2024-07-01 05:26:32","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":42301,"visible":true,"origin":"","legend":"\u003cp\u003eDPPH inhibition effect of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and its self-nanoemulsifying formulation.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4467611/v1/3bb8da8a512528d9858ba098.png"},{"id":59380514,"identity":"fda75d2f-9acb-40f9-bd1f-5c6b22e84f85","added_by":"auto","created_at":"2024-07-01 05:26:32","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":37171,"visible":true,"origin":"","legend":"\u003cp\u003eα-amylase\u003cstrong\u003e \u003c/strong\u003einhibition effect of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and its self-nanoemulsifying formulation\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4467611/v1/6286099d87aee86adb3e5f31.png"},{"id":59380519,"identity":"ccf016d6-c495-46c4-8cae-4e1c69860a74","added_by":"auto","created_at":"2024-07-01 05:26:32","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":39522,"visible":true,"origin":"","legend":"\u003cp\u003elipase inhibition effect of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and its self-nanoemulsifying formulation.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-4467611/v1/aaf2bfd87259f7109ee142a5.png"},{"id":63640580,"identity":"491e7f7c-7003-419b-b9ed-c81cbdf6fb75","added_by":"auto","created_at":"2024-08-30 12:52:38","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1218619,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4467611/v1/53e2e5f3-c679-4584-a010-606205770399.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Phytochemical and Biological Evaluation of a Newly Designed Vitis Vinifera Seed Oil Self-Nanoemulsifying System","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eA wide range of therapeutic plants exist in the world. Prehistoric human beings stumbled upon the healing properties of medicinal plants through trial and error in their quest to alleviate disease symptoms. The general consensus is that natural remedies are safer, more benign, and healthier than synthetic ones. Therefore, many diseases have been treated with medicinal plants [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn the past, natural chemicals and similar structures have been important in pharmacology, especially for treating infectious disorders and cancer. There are around 100 new products now undergoing clinical development particularly for these disorders [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. In order to facilitate the utilization of natural products in drug discovery efforts, many screening methodologies are being devised. Data mining and virtual screening approaches are being used to databases that include information on natural products. Utilizing natural materials in a more effective and efficient manner is anticipated to improve the process of discovering new drugs [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. The swift progress of technology has led to the emergence of innovative methods in the quest for pharmaceuticals obtained from natural sources. The utilization of artificial intelligence and bioinformatics has facilitated the examination and production of natural products [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eVitis vinifera\u003c/em\u003e (\u003cem\u003eV. vinifera\u003c/em\u003e) is a perennial plant belonging to the \u003cem\u003evitaceae\u003c/em\u003e family. The primary bioactive components found in \u003cem\u003eV. vinifera\u003c/em\u003e seed oil are phenolic compounds, which encompass flavonoids, tannins, phenolic acids, carotenoids, and stilbenes. Grape seed oil contains phenolic components such as catechins, epicatechins, and gallic acid, as well as fatty acids including linoleic acid, vitamin E, and phytosterols. The seeds have an elliptical or pyriform morphology with a tapered apex. Their length can extend up to 6 mm [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eV. vinifera\u003c/em\u003e seed possesses antioxidant, antifungal, antidiabetic, anti-inflammatory, and anti-obesity properties, as well as exhibiting anticholinergic scarring effects. \u003cem\u003eV. vinifera\u003c/em\u003e seed oil includes essential components such as ascorbic acid, beta-carotene, phytosterols, and tocopherols. These chemicals, which are present in the stems and leaves of the plant, provide protection against oxidative stress. Goufo, Singh, and Cortez (2020) found that the presence of proanthocyanidins in the seeds is indicative of the oil's antioxidant potential. Scientific study has demonstrated that \u003cem\u003eV. vinifera\u003c/em\u003e seed possess distinct biological and therapeutic characteristics, making them advantageous for one's health. However, a limited number of research that employ experimental models of obesity, oxidative stress, or inflammation document the therapeutic capacity of these models in treating metabolic diseases [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eResearch has shown that \u003cem\u003eV. vinifera\u003c/em\u003e seed oil effectively inhibits the rapid development of human cancer cells. Grape seed oil contains organic fatty acids, including malic, tartaric, and oxalic acids, which have the ability to reduce LDL-c levels [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. \u003cem\u003eV. vinifera\u003c/em\u003e seed oil is rich in tocotrienols, particularly α- and γ-tocotrienol (T3), which effectively decrease the production of mRNA protein. This reduction is crucial for inhibiting the process of adipogenesis, namely the activity of PPARγ and aP2. Furthermore, it substantially reduces the production of pro-inflammatory genes, including IL-6 and IL-8 [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. The presence of polyphenols, including flavanols, catechin, and epicatechin, in this function further improves its effectiveness. These polyphenols may be associated with preventing arterial thrombosis. Additionally, compounds such as flavones, isoflavones, and anthocyanins are involved in preventing pathologies related to weight gain and possess anti-inflammatory properties. [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eV. vinifera\u003c/em\u003e has been used as a key component in the cosmetic sector due to its significant antioxidant, antibacterial, and skin care attributes [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. he CosIng (cosmetic ingredients) database states that cosmetics utilize grape-derived compounds in nine different ways [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. The extract of \u003cem\u003eV. vinifera\u003c/em\u003e can serve as an emollient, humectant, emulsifier, color addition, or fragrance. For instance, grape seeds are suggested for use as a color, moisturizer, or hair and skin care product. Grape-derived cosmetics possess gentle and non-aggravating properties when applied to the skin. Scientific study indicates that this plant possesses antioxidant, whitening, anti-inflammatory, anti-aging, and other properties mostly because of its abundant phenolic and stilbene constituents. It is worth noting that \u003cem\u003eV. vinifera\u003c/em\u003e oil or extracts has distinctive moisturizing and anti-aging properties, as evidenced by the increasing use of these fundamental constituents in cosmetic goods [\u003cspan additionalcitationids=\"CR15\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAccording to Wijekoon et al., ingesting grape varietals with seeds is important as a functional diet since they contain considerable amounts of anthocyanins, flavones, flavonols, and stilbenes (specifically resveratrol) [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Argon et al. state that grape seed oil includes bioactive chemicals such as carotenoids, tocopherols (tocols), phytosterols, phenolic components, and other fat-soluble compounds. Tocopherols are powerful, naturally-occurring antioxidants that dissolve in fats [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. An experiment conducted on mice revealed that those mice who were fed a hyperlipidic diet enriched with 15.5 g of grape seed oil per 100 g of food for a duration of eight weeks had reduced levels of glucose and total cholesterol in their blood plasma, as well as enhanced glucose tolerance. Furthermore, the subjects exhibited reduced weight gain and decreased weight of white adipose tissue. Additionally, they demonstrated significant antioxidant properties and lower levels of TBARS in their plasma, which are indicators of lipid peroxidation. Moreover, there was a decrease in the production of IL-6 and IL-10. Based on the evidence, it can be inferred that V. vinifera seed oil is classified as a functional oil that has the capacity to decrease inflammation and obesity issues caused by oxidative stress [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eNanotechnology refers to the manipulation and creation of materials at extremely small sizes, namely at the atomic and molecular levels [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Scientists are developing novel formulations such as nanoemulsions [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e] to address the limitations of existing formulations, which include inadequate targeting of particular sites, poor spreading ability, and insufficient bioavailability. The nanoemulsion (NE) system is characterized by its stability, viscous formulation, tiny particle size, high encapsulation efficiency, good thermodynamic stability, and strong penetration ability [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Nanoemulsions consist of droplets with a size range of 10 to 100 nm, allowing them to quickly enter and distribute active ingredients profoundly [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Self-emulsifying drug delivery methods are crucial for resolving the issue of low bioavailability associated with drugs that have poor solubility [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Repeatedly, the use of nutraceuticals has been linked to safeguarding against chronic ailments such as diabetes, heart disease, cancer, and neurological disorders. However, the therapeutic potential of these nutraceuticals is limited due to their low permeability and poor solubility in water. Therefore, the utilization of self-emulsifying formulation has demonstrated promising potential in improving the absorption rate and effectiveness of nutraceuticals, addressing the aforementioned challenges [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. This work aims to extract, analyze the phytochemical characteristics, and assess the biological activities of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and its self-nanoemulsifying formulation.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Materials\u003c/h2\u003e \u003cp\u003eDimethyl sulfoxide (DMSO) was obtained from Riedel De Haen, Germany, and p-nitrophenyl butyrate (PNPB) from Sigma-Aldrich, Germany. Sigma-Aldrich (USA) provided acarbose, porcine pancreatic lipase type 2, starch, orlistat, (p-nitrophenyl α-D-galactopyranoside) PNPG, and 2,2-diphenyl-1-picrylhydrazyl (DPPH). Both Span 80 and Tween 80 were obtained from the Al-Shams company (Palestine). Trolox [(S)-(-)-6-hydroxy-2, 5, 7, 8-tetramethylchroman-2-carboxyylic acid]. The α-amylase enzyme from Sigma-Aldrich (India) and n-hexane (Frutarom Ltd., Israel).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Extraction of the \u003cem\u003eV. vinifera\u003c/em\u003e seed essential oil\u003c/h2\u003e \u003cp\u003eThe \u003cem\u003eV. vinifera\u003c/em\u003e fruits were collected in 2023 from Hebron in Palestine. The fruits were then identified at the pharmacognosy laboratory at An-Najah National University using the voucher specimen code Pharm-PCT-2665. The fruit seeds were dried in the shade at a controlled temperature [25\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C] and humidity [55\u0026thinsp;\u0026plusmn;\u0026thinsp;4 RH]. Following the drying process, we ground the seeds into a fine powder. About 50 g of the powdered \u003cem\u003eV. vinifera\u003c/em\u003e seeds were soaked in 250 mL of 70% ethanol, a polar solvent, and 250 mL of hexane, a non-polar organic solvent, for 72 hours in a shaker at room temperature. Afterwards, we divide the mixture into two layers using a separotary funnel: the upper layer, the organic phase containing the oil, and the lower layer, the aqueous phase. Then the solvent was removed under air pressure to concentrate the organic layer [\u003cspan additionalcitationids=\"CR28\" citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Bioactive components study of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil by GC-MS\u003c/h2\u003e \u003cp\u003eUsing the gas chromatography-mass spectrometry (GC-MS) method, the fatty acid methyl ester (FAME) of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil was analyzed. To prepare the oil, we mixed 100 mg of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil with 10 ml of a 0.5 M methanolic sodium hydroxide solution, then heated the mixture for complete hydrolysis on a heating mantel for 10 minutes. After that, I added 10 ml of 14% boron trifluoride (BF3) solution in methanol and refluxed them for 10 minutes. After the esterification, add 50 ml of heptan and continue the boiling for 10 minutes, then the mixture was cooled. A Perkin Elmer Clarus 500 gas chromatograph and a Perkin Elmer Clarus 560 mass spectrometer were used to examine the FAME of V. vinifera seed oil in the heptan layer. A fused silica capillary column (30 m x 0.25 mm, film thickness 0.25 \u0026micro;m) was used. The column temperature was planned to increase by 4\u0026deg;C/min, so the temperature of the well increased from 3\u0026deg;C to 240\u0026deg;C (hold for 15 minutes). Helium gas (99.999%) is used as a carrier with a flow rate of 1 ml/min. With a split ratio of 50:1 and at a temperature of 25\u0026deg;C, 1 \u0026micro;l of oil was injected in split mode. Solvent delay: 0\u0026ndash;8 min. The entirety of the data was obtained through the collection of full-scan mass spectra spanning the range of 50\u0026ndash;500 atomic mass units (amu). Ionizing energy measured 70 eV. The voltage of the electron multiplier (EM) was acquired via autotune. The mass spectrum of the chemical components of \u003cem\u003eV. vinifera\u003c/em\u003e oil was compared to the reference spectrum from the MS Database of the National Institute of Standards and Technology, and their retention and kovats results were compared to other research results [\u003cspan additionalcitationids=\"CR28\" citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4. Preperation of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil nanoemulsion\u003c/h2\u003e \u003cp\u003eUsing the self-nanoemulsifying technique, we convert the extracted \u003cem\u003eV. vinifera\u003c/em\u003e seed oil into a nanoemulsion following extraction. To construct the ternary phase diagram, three different substances were used at varying concentrations: \u003cem\u003eV. vinifera\u003c/em\u003e seed oil, Span 80 as a co-surfactant, and Tween 80 as a surfactant! After determining the weight of each formulation using a weighting balance, the mixture was agitated gently for three minutes with a vortex mixer in order to achieve homogeneity. Then, each formulation was emulsified under gentle agitation with distilled water. The formulation that yields the highest possible particle size and polydispersity index (PDI) has been determined [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5. Droplet size and PDI analysis of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil nanoemulsion\u003c/h2\u003e \u003cp\u003eTo determine the droplet size and PDI, a master size analyzer (Brookhaven Instruments, NanoBrook Omni, New York) was used. The \u003cem\u003eV. vinifera\u003c/em\u003e seed oil nanoemulsion was self-emulsified in distilled water before using the measuring procedure. The ideal nanoemulsion formulation was chosen based on the smallest droplet size and PDI and with the highest amount of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.6. Antioxidant activity of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil\u003c/h2\u003e \u003cp\u003eInitially, methanol was used to produce stock solutions (1 mg/ml) of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and its self-nanoemulsifying formulation. At the following concentrations (2, 5, 10, 20, 50, 80, and 100 \u0026micro;g/ml), each of the stock solutions was diluted in methanol. A DPPH solution that had been recently prepared was dissolved in methanol at a concentration of 0.002% w/v. Following that, a 1:1:1 ratio was maintained between each working solution and methanol and DPPH, with the lowest concentration added last. The solutions were incubated in a dark area at room temperature for thirty minutes. As a baseline, methanol was utilized on the UV-visible spectrophotometer; all absorbance values were calculated at a wavelength of 517 mm. Trolox was employed in the capacity of a positive control. Determine the percentage of antioxidant activity exhibited by each tested compound and the Trolox standard by employing the subsequent equation:\u003c/p\u003e \u003cp\u003e[\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e% DPPH inhibition = (A\u003csub\u003eb\u003c/sub\u003e \u0026ndash; A\u003csub\u003es\u003c/sub\u003e ) / A\u003csub\u003eb\u003c/sub\u003e \u0026times; 100%\u003c/p\u003e \u003cp\u003eA\u003csub\u003eb\u003c/sub\u003e: blank absorbance, and A\u003csub\u003es\u003c/sub\u003e: sample absorbance.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.7. Anti-diabetic activity of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil using α-amylase inhibitory method\u003c/h2\u003e \u003cp\u003eThe inhibitory effect of the α-amylase enzyme by \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and its self-nanoemulsifying formulation was evaluated using a modified version of a standard technique that had been previously established [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. In order to achieve a concentration of 1000 \u0026micro;g/mL, the oil and its self-nanoemulsion in 10% DMSOO were dissolved individually in a buffer solution consisting of NaCl and Na2HPO4. Several dilutions containing 10, 50, 70, 100, and 500 \u0026micro;g/mL were subsequently performed. After combining 0.2 mL of the oil with an equivalent volume of panreatic amylase enzyme solution, the mixture was incubated at 30\u0026deg;C for a duration of 10 minutes. The mixture was then incubated for three minutes after 0.2 mL of starch solution was added. 0.2 mL of DNSA is introduced to halt the incubation process. The mixture was then diluted with 5 mL of distilled water and heated for 10 minutes at 90\u0026deg;C in a water bath. 540nm absorption measurements were taken of the mixtures subsequent to their reduction to room temperature. To produce the blank, a buffer solution was substituted for the oil. For the positive control, which is a carbohydrate, the exact same procedure steps were followed. Using the equation below, The inhibitory activity of α-amylase was assessed [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e% α- amylase inhibition = (A\u003csub\u003eb\u003c/sub\u003e \u0026ndash; A\u003csub\u003es\u003c/sub\u003e ) / A\u003csub\u003eb\u003c/sub\u003e \u0026times; 100%\u003c/p\u003e \u003cp\u003eA\u003csub\u003eb\u003c/sub\u003e: blank absorbance, and A\u003csub\u003es\u003c/sub\u003e: tested sample absorbance.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e2.8. Anti-obesity acivity of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil using lipase inhibitory method\u003c/h2\u003e \u003cp\u003eStock solutions of orlistat, a positive control, and \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and its self-nanoemulsion were prepared at a concentration of 1000 \u0026micro;g/mL. Subsequently, the solutions mentioned above are diluted in a test tube containing 50, 100, 200, 300, and 400 \u0026micro;g/mL. A bulk solution of pancreatic lipase enzyme was formulated in a 10% DMSO solution. Following that, 0.1 mL of pancreatic lipase enzyme and 0.7 mL of buffer containing Tris-HCl were added to the solution. Each test tube, along with a blank tube containing Tris-HCl and pancreatic lipase enzyme, was incubated at 37\u0026deg;C for a duration of 15 minutes. Additional incubation for 30 minutes in the blank and each test tube subsequent to adding 0.1 mL of PNPB. In order to evaluate the hydrolysis of PNPB to p-nitophenol, a 410 nm UV-visible spectrometer was employed [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e% lipase inhibition = (A\u003csub\u003eb\u003c/sub\u003e \u0026ndash; A\u003csub\u003es\u003c/sub\u003e ) / A\u003csub\u003eb\u003c/sub\u003e \u0026times; 100%\u003c/p\u003e \u003cp\u003eA\u003csub\u003eb\u003c/sub\u003e: blank absorbance, and A\u003csub\u003es\u003c/sub\u003e: sample absorbance.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Analysis of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil chemical components using GC-MS\u003c/h2\u003e \u003cp\u003eThe biologically active components of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil were detected using a GC-MS device, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The main components showed in Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e were linoleic acid methyl ester (62.97%), oleic acid methyl ester (23.07%), palmitic acid methyl ester (8.36%), stearic acid methyl ester (4.96%), and elaidic acid methyl ester (0.64%).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eNatural active compounds characterized by GC-MS of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil\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\u003eFatty acid of grape seed oil\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e% age\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePalmitic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.36%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLinoleic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.97%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOleic Acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e23.07%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eElaidic Acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.64%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStearic Acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.96%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e100%\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=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Droplet size and PDI analysis of \u003cem\u003eVitis Vinifera\u003c/em\u003e nanoemulsion formulations\u003c/h2\u003e \u003cp\u003eIn order to determine the most effective formulation, ternary phase diagrams were constructed by employing different concentrations of Span 80, Tween 80, and \u003cem\u003eV. vinifera\u003c/em\u003e seed oil (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Subsequently, a nanoemulsion was generated characterized by a droplet size below 200 nm and a PDI below 0.2. In order to conduct a comparative analysis of the chosen formulations exhibiting droplet sizes below 200 nm, we employed the optimal nanoemulsion formulation consisting of 35% \u003cem\u003eV. vinifera\u003c/em\u003e seed oil, 50% Tween 80, and 15% Span 80 (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). This formulation exhibited a PDI of 0.227\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07 and a droplet size of 192.71\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8 nm.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe selected optimum \u003cem\u003eVitis Vinifera\u003c/em\u003e seed oil self-nanoemulsifying formulation.\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\u003eTween 80(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSpan 80 (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOil(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDroplet size(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePolydispersity Index(PDI)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e192.71\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.227\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\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=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e3.3. Antioxidant activity of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and its self-nanoemulsifying formulation\u003c/h2\u003e \u003cp\u003eThe DPPH radical method was employed to determine the free radical scavenging activity of the extracted \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and it self-nanoemulsifying formulation, with trolox serving as positive control. The prospective antioxidant activity of the essential oil of \u003cem\u003eV. vinifera\u003c/em\u003e was demonstrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, where it exhibited an IC\u003csub\u003e50\u003c/sub\u003e value of 33.88\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7 \u0026micro;g/ml, in contrast to the positive control (Trolox) which showed an IC\u003csub\u003e50\u003c/sub\u003e value of 4.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6 \u0026micro;g/ml. The oil possesses a lower inhibitory activity than the self-nanoemulsion, as evidenced by its IC\u003csub\u003e50\u003c/sub\u003e value of 13.80\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5 \u0026micro;g/ml.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e\u003cb\u003e3.4. Anti-diabetic activity\u003c/b\u003e \u003cb\u003eof V. vinifera\u003c/b\u003e \u003cb\u003eseed oil and its self-nanoemulsifying formulation\u003c/b\u003e\u003c/h2\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e shows the percentage of α-amylase inhibition for \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and its self-nanoemulsifying formulation at various concentrations compared to acarbose as a positive control. The IC\u003csub\u003e50\u003c/sub\u003e values for \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and its self-nanoemulsifying formulation were 257.03\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4 \u0026micro;g/ml and 52.48\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2 \u0026micro;g/ml, respectively. These values were not as strong as the IC\u003csub\u003e50\u003c/sub\u003e value for acarbose, which was 39.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8 \u0026micro;g/ml. The oil activity improved when it was converted to a self-nanoemulsifying system and showed an IC\u003csub\u003e50\u003c/sub\u003e close to that of the positive control.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003e3.5. Anti-obesity activity \u003cem\u003eof V. vinifera\u003c/em\u003e seed oil and its selfnanoemulsifying formulation\u003c/h2\u003e \u003cp\u003e \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and its self-nanoemulsifying formulation exhibited porcine lipase inhibition; this was subsequently evaluated in comparison to orlistat, which serves as the positive control. The lipase inhibitory activity of the oil is significantly high when compared to that of orlistat, as illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. IC\u003csub\u003e50\u003c/sub\u003e values for the oil and orlistat were 33.88\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7 \u0026micro;g/ml and 12.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8 \u0026micro;g/ml, respectively. With an IC\u003csub\u003e50\u003c/sub\u003e value of 13.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5 \u0026micro;g/ml, the self-nanoemulsifying formulation of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil exhibits superior inhibitory activity in comparison to the oil, specifically orlistat.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eThe chemical constituents of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil were identified using a Gas Chromatography-Mass Spectrometry (GC-MS) instrument, as seen in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The primary constituents were of linoleic acid methyl ester (62.97%), oleic acid methyl ester (23.07%), palmitic acid methyl ester (8.36%), stearic acid methyl ester (4.96%), and elaidic acid methyl ester (0.64%). The study conducted by Al-Fekaiki and Al-Hilfi. The fatty acid composition of V. vinifera seed oil was determined in 2016 using GC-MS. The analysis revealed the presence of various fatty acids, including oleic acid (16.98%), palmitic acid (12.09%), stearic acid (8.85%), palmitoleic acid (0.28%), myristic acid (0.21%), azelaaldehydic acid (0.12%), margaric acid (0.11%), lauric acid (0.05%), and pentadecanoic acid (0.03%) [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eA recent study conducted by Sabir, Unver, and Kara investigated the composition of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and found varying percentages of different fatty acids. The major component was linoleic acid, accounting for 53.6\u0026ndash;69.6% of the oil. This was followed by oleic acid (16.2\u0026ndash;31.2%), palmitic acid (6.9\u0026ndash;12.9%), and stearic acid (1.44\u0026ndash;4.69%). Additionally, we discovered many chemical constituents such as alpha-tocopherol, beta-tocopherol, gamma-tocopherol, and sigma-tocopherol. The oil samples from grape seeds include much larger quantities of linoleic acid and substantial levels of tocopherols, indicating that grape seed is a valuable source for culinary, medicinal, and cosmetic applications [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. A further investigation on \u003cem\u003eV. vinifera\u003c/em\u003e seed oil revealed a substantial quantity of phenolic substances, encompassing flavonoids, carotenoids, phenolic acid, tannins, and stilbenes. Gallic acid possesses antioxidant properties and contains polyphenols such as catechins, epicatechins, trans-resveratrol, and procyanidin B1 [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eBy creating formulations with varying concentrations of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil, surfactant, and co-surfactant, we constructed a ternary phase diagram to identify the optimal formulation for the self-nanoemulsifying drug delivery system (SNEDDS). The goal was to achieve a droplet size smaller than 200 nm and a PDI (polydispersity index) lower than 0.2. Tween 80 and Span 80 are non-ionic surfactants that make up 50% and 15% of our composition, respectively. Multiple studies have demonstrated a direct correlation between an increase in surfactant content and a decrease in droplet size. Greater quantities of surfactant are needed to stabilize smaller droplets since they have a larger surface area [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. Moreover, a higher quantity of surfactant is necessary to stabilize diminutive particles that possess a larger surface area. Recent study suggests that an elevation in the surfactant concentration in our formulation might result in several outcomes, such as a reduction in surface tension and an expansion in artificial area, possibly due to an enlargement in particle size [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. Singh et al. (2017) shown that nanoemulsion systems are highly effective in enhancing medication bioavailability, delivery, solubility, stability, and effectiveness [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eA self-nanoemulsifying system was created to boost the bioactivity of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil, resulting in greater solubility and hence higher efficacy [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. Wang et al. (2023) conducted a research to examine the advantageous characteristics of lingonberry leaves and their active constituents. The study revealed that these features were further increased when included into nanoemulsions. The study's findings were acquired concurrently using internet means. The purpose of developing nanoemulsions was to enhance the stability and antioxidant effectiveness of extracts from lingonberry leaves. The nanoemulsions exhibited enhanced antioxidant activity, with a DPPH radical inhibition rate exceeding 13% at the same dose. This enhancement was statistically significant when compared to their non-nanoemulsified counterparts. The study discovered that nanoscale emulsions greatly enhance the solubility of polyphenols, leading to the optimization of their antioxidant capabilities [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eMultiple studies have shown that grape seeds possess strong antioxidant properties, possibly attributed to the presence of different phenolic compounds such as epicatechin, catechin, procyanidins, proanthocyanidins, gallic acid, and ellagic acid [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. Several investigations have demonstrated that \u003cem\u003eV. vinifera\u003c/em\u003e seed oil exhibits other actions beyond its antioxidant properties. These include anti-inflammatory, anti-cancer, anti-microbial, anti-ulcer, and cardioprotective benefits [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. The research conducted by P\u0026eacute;rez et al. (2015) confirmed that grape seed oil extract had remarkable antioxidant properties, with a scavenging activity of 95.8% against DPPH radicals [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. Our results were consistent with this finding, as the \u003cem\u003eV. vinifera\u003c/em\u003e seed oil exhibited noteworthy antioxidant properties. The IC\u003csub\u003e50\u003c/sub\u003e value for the \u003cem\u003eV. vinifera\u003c/em\u003e seed oil was 33.88\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7 \u0026micro;g/ml, while the self-nanoemulsifying system had an IC\u003csub\u003e50\u003c/sub\u003e value of 13.80\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5 \u0026micro;g/ml, indicating even stronger antioxidant activity. In comparison to trolox, which had an IC\u003csub\u003e50\u003c/sub\u003e value of 4.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6 \u0026micro;g/ml, the self-nanoemulsifying system demonstrated superior antioxidant activity compared to the pure oil.\u003c/p\u003e \u003cp\u003eObesity and diabetes are becoming prevalent health problems worldwide. They have been recognized as major risk factors for many infections, complications following infections, and death caused by severe infections [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. The study conducted by Ahmed et al. investigated the antidiabetic effects of grape stem extract. The results showed that both the chloroform and ethanolic stem extracts were able to lower blood glucose levels in a dose-dependent way. The maximum activity was shown at a dosage of 200 mg/kg bodyweight [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. The IC\u003csub\u003e50\u003c/sub\u003e value for \u003cem\u003eV. vinifera\u003c/em\u003e seed oil was determined to be 257.03\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4 \u0026micro;g/ml, while the IC\u003csub\u003e50\u003c/sub\u003e value for the oil self-nanoemulsifying system was found to be 33.88\u0026thinsp;\u0026plusmn;\u0026thinsp;1.6 \u0026micro;g/ml. These values, although relatively moderate, were compared to the reference compound Acarbose, a potent α-amylase inhibitory agent, which exhibited an IC\u003csub\u003e50\u003c/sub\u003e value of 39.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8 \u0026micro;g/ml. The aqueous extracts of \u003cem\u003eV. vinifera\u003c/em\u003e exhibited the most potent anti-lipase activities, as indicated by their IC\u003csub\u003e50\u003c/sub\u003e values of 14.13\u0026micro;g/ml. \u003cem\u003eV. vinifera\u003c/em\u003e exhibited inhibitory effects on pig pancreatic lipase compared to Orlistat, with an IC\u003csub\u003e50\u003c/sub\u003e value of 12.38 \u0026micro;g/ml. This work was performed on \u003cem\u003eV. vinifera\u003c/em\u003e and its self-nanoemulsifying system. The aqueous extracts of \u003cem\u003eV. vinifera\u003c/em\u003e exhibited strong anti-lipase actions, with IC\u003csub\u003e50\u003c/sub\u003e values of 14.13 \u0026micro;g/ml. Concurrently, \u003cem\u003eV. vinifera\u003c/em\u003e exhibited inhibitory effects on pig pancreatic lipase compared to orlistat, with an IC\u003csub\u003e50\u003c/sub\u003e value of 12.38 \u0026micro;g/ml [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. A nanoemulsion was created to enhance the bioactivity of \u003cem\u003eV. vinifera\u003c/em\u003e seed oil. The improvement occurred because the nanoemulsion had a higher solubility compared to the oil and other drugs. This is because the nanoemulsion has a larger surface area and very small particle size, which both contribute to its increased effectiveness [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e].\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eThe present study investigated the anti-diabetic, antioxidant, and anti-obesity effects of \u003cem\u003eV. vinifera\u003c/em\u003e nanoemulsion in comparison to pure oil and positive control drugs. The undiluted oil exhibited anti-obesity effects against lipase, antioxidant effects against DPPH, and anti-diabetic effects against α-amylase; these effects were enhanced when the oil was in the form of a nanoemulsion. In comparison to the unadulterated \u003cem\u003eV. vinifera\u003c/em\u003e seed oil, the nanoemulsion exhibited noteworthy antioxidant, anti-diabetic, and anti-obesity properties, a reduction in droplet size, and an enhancement in stability and solubility. The results indicate that \u003cem\u003eV. vinifera\u003c/em\u003e seed oil is a valuable reservoir of bioactive and antioxidant substances, particularly unsaturated fatty acids such as linoleic acid, palmitic acid, oleic acid, and stearic acid. This study proposes that incorporating \u003cem\u003eV. vinifera\u003c/em\u003e seed oil into nanoemulsions can be a highly effective method for formulating oral medications. Hence, owing to its capacity to hinder the activity of α-amylase, lipase, and free radicals in comparison to \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and Trolox, it has the potential to be employed as a supplementary treatment for obesity and diabetes, alongside other drugs, in order to impede the advancement of these disorders. Furthermore, as demonstrated by this work, a self-nanoemulsion system may serve as a reservoir for drugs, improve the way drugs are absorbed and distributed in the body, and facilitate the release of oil in oil-based therapeutic formulations.\u003c/p\u003e "},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003ePermissions and identification of plant material\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePermissions were obtained to collect the plant material. The plant material was collected and\u0026nbsp;identified within the Pharmacy Department of An-Najah National University.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAuthors confirm that the used of plant in the present study, including the collection of plant material, comply with relevant institutional, national, and international guidelines and legislation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe manuscript file contains all of the data that was created or analyzed for this project. I confirm that the study has all the information and consents required.\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\u003eNone.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAE: conceptualization, formal analysis, supervision, writing\u0026mdash;original draft. LI, HS, RS, SM, and TH carried out the formal analysis,\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to acknowledge the Faculty of Medicine and Health Sciences at An-Najah National University.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eMannangatti P, Naidu KN. 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Antidiabetic, antioxidant, and anti-obesity effects of phenylthio-ethyl benzoate derivatives, and molecular docking study regarding α-amylase enzyme. Sci Rep. 2022;12(1):3108.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAl-fekaiki D, Ali S. Fatty Acids Composition by (GC-MS) and Most Important Physical Chemicals Parameters of Seed Oil Pomegranate and Grape Seeds2016.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSabir A, Unver A, Kara Z. The fatty acid and tocopherol constituents of the seed oil extracted from 21 grape varieties (Vitis spp). J Sci Food Agric. 2012;92(9):1982\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePolitova NI, Tcholakova S, Tsibranska S, Denkov ND, Muelheims K. Coalescence stability of water-in-oil drops: Effects of drop size and surfactant concentration. 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Foods. 2023;12(23):4256.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"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":"Vitis Vinifera, self-nanoemulsifying system, Phytochemical, Antioxidant, Anti-diabetic, anti-lipase","lastPublishedDoi":"10.21203/rs.3.rs-4467611/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4467611/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eThe objective of this research is to extract and formulate a nanoemulsion consisting of \u003cem\u003eVitis vinifera\u003c/em\u003e (\u003cem\u003eV. vinifera\u003c/em\u003e) oil. Following this, the antioxidant, anti-diabetic, and anti-lipase properties of both the oil and the nanoemulsion system will be examined.\u003c/p\u003e\u003ch2\u003eMethod\u003c/h2\u003e \u003cp\u003e \u003cem\u003eV. vinfera\u003c/em\u003e seed oil extraction was followed by the preparation of its self-nanoemulsifying system. GC analysis was conducted to analyze phytochemical compounds, and physical characterization was performed for the \u003cem\u003eV. vinfera\u003c/em\u003e oil self-nanoemulsifying system. Subsequently, antioxidant, anti-diabetic, and anti-lipase activities were investigated for the oil and its nanoemulsion.\u003c/p\u003e\u003ch2\u003eResult\u003c/h2\u003e \u003cp\u003eThe biologically active components of \u003cem\u003eV. vinifera\u003c/em\u003e oil were detected using a GC/MS device. The main components were: linoleic acid methyl ester, oleic acid methyl ester, palmitic acid methyl ester, and stearic acid methyl ester, and elaidic acid methyl ester. The optimum nanoemulsion formulation, which comprised Tween 80, Span 80, and \u003cem\u003eV. vinifera\u003c/em\u003e seed oil and had a PDI of 0.227\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07 and a droplet size of 192.71\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8 nm. Porcine lipase inhibition of \u003cem\u003eV. vinifera\u003c/em\u003e oil and its nanoemulsion was detected; the oil showed potent lipase inhibitory activity when compared to orlistat. The self-nanoemulsion has less inhibitory activity than the oil, with an IC\u003csub\u003e50\u003c/sub\u003e equivalent to 13.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5 \u0026micro;g/ml. Also, \u003cem\u003eV. vinifera\u003c/em\u003e oil inhibited a-amylase with a weak IC\u003csub\u003e50\u003c/sub\u003e value of 257.03\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4 \u0026micro;g/ml. The free radical scavenging activity of the extracted \u003cem\u003eV. vinifera\u003c/em\u003e oil was estimated using the DPPH radical method and trolox. The self-nanoemulsion has less inhibitory activity than the oil, with an IC\u003csub\u003e50\u003c/sub\u003e value of 13.80\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5 \u0026micro;g/ml.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eThis study demonstrated the importance of black grape (\u003cem\u003eV. vinfera\u003c/em\u003e) oil for treating and preventing obesity, diabetes, and oxidative stress, as well as how nanoemulsions improve these activities. Therefore, this natural oil is a promising product for the pharmaceutical industry.\u003c/p\u003e","manuscriptTitle":"Phytochemical and Biological Evaluation of a Newly Designed Vitis Vinifera Seed Oil Self-Nanoemulsifying System","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-01 05:26:27","doi":"10.21203/rs.3.rs-4467611/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":"e1e56ed5-c579-4e85-a1fb-7a4dc55bc838","owner":[],"postedDate":"July 1st, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-08-30T12:44:31+00:00","versionOfRecord":[],"versionCreatedAt":"2024-07-01 05:26:27","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4467611","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4467611","identity":"rs-4467611","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}