In Vitro and In-Vivo Analysis of the Combined Extract of Tabernaemontana divaricata and Mangifera indica Leaves for Enhanced Anti-Inflammatory and Antioxidant Properties

In Vitro and In-Vivo Analysis of the Combined Extract of Tabernaemontana divaricata and Mangifera indica Leaves for Enhanced Anti-Inflammatory and Antioxidant Properties | 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 In Vitro and In-Vivo Analysis of the Combined Extract of Tabernaemontana divaricata and Mangifera indica Leaves for Enhanced Anti-Inflammatory and Antioxidant Properties Ganesh Mangesh Walanj, Anand Kumar, Dilip KumarTiwari, Abhishek Sharma, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5344453/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 Ethnopharmacological Relevance: Tabernaemontana divaricata ( T. divaricata ) and Mangifera indica ( M. indica ) are plants traditionally used in Ayurvedic medicine for their anti-inflammatory and antioxidant properties. T. divaricata is known for its analgesic and wound-healing effects, while M. indica is celebrated for its role in managing inflammation and oxidative stress. Aim of the Study: The study aimed to evaluate the synergistic effects of combined hydroalcoholic extracts from T. divaricata and M. indica leaves, focusing on their anti-inflammatory and antioxidant properties. Materials and Methods The leaves were collected, authenticated, and subjected to hydroalcoholic extraction. The extracts were then combined in various ratios and analyzed for phytochemical content. The antioxidant activity was evaluated using the DPPH assay, while anti-inflammatory effects were assessed through the carrageenan-induced rat paw edema model. Results The combined extracts exhibited significantly enhanced antioxidant and anti-inflammatory activities compared to the individual extracts. The most potent combination, a 2:1 ratio of M. indica to T. divaricata , showed the highest DPPH inhibition (94.37%) and maximum edema reduction in the animal model. Conclusion The combination of T. divaricata and M. indica extracts offers a synergistic therapeutic potential, enhancing both antioxidant and anti-inflammatory effects, which supports their traditional use and suggests potential for developing new natural therapeutics. Clinical Pharmacology Anti-inflammatory Anti-oxidant Tabernaemontana divaricate Mangifera indica Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 1. Introduction Inflammation and oxidative stress are two pivotal processes implicated in the pathogenesis of various chronic diseases, including cardiovascular disorders, cancer, diabetes, and neurodegenerative conditions (Leyane, Jere, and Houreld 2022 ). The increasing prevalence of these conditions has heightened the search for natural remedies that can offer effective therapeutic interventions with minimal side effects. In this context, plants and their bioactive compounds have garnered significant attention due to their historical use in traditional medicine and their potential in modern therapeutics (Pandey et al. 2011 ). Tabernaemontana divaricata , commonly known as the crepe jasmine (Ghosh, Poddar, and Chatterjee 2021). Tabernaemontana divaricata belongs to the family Apocynaceae. This family is commonly known as the dogbane family, and it includes a wide variety of flowering plants, many of which have medicinal properties. Tabernaemontana divaricata is widely used in Ayurvedic medicine, primarily for its anti-inflammatory, analgesic, and wound-healing properties. The plant is rich in alkaloids, flavonoids, and terpenoids, which contribute to its therapeutic effects. Mangifera indica , known as the mango tree (Ghosh, Poddar, and Chatterjee 2021) is the plants traditionally recognized for their medicinal properties. Mangifera indica belongs to the family Anacardiaceae, which is known for including several economically important fruit-bearing trees and plants. Mangifera indica , on the other hand, is renowned not only for its nutritional value but also for its extensive medicinal applications (Ediriweera, Tennekoon, and Samarakoon 2017), particularly in treating inflammation, infections, and oxidative stress-related conditions. The leaves of Mangifera indica are known to contain polyphenols, flavonoids, and other potent antioxidants (Mirza et al. 2021 ). Given the individual therapeutic properties of these plants, combining the extracts of Tabernaemontana divaricata and Mangifera indica leaves presents a promising approach to enhancing their anti-inflammatory and antioxidant effects. The rationale behind this combination is to leverage the synergistic effects of the bioactive compounds present in both plants, which may result in a more potent therapeutic outcome than the individual extracts. This study aims to explore the anti-inflammatory and antioxidant properties of the combined extract of Tabernaemontana divaricata and Mangifera indica leaves. By evaluating these effects through various in vitro and in vivo models, the research seeks to establish a scientific basis for the traditional use of these plants and potentially contribute to the development of new natural therapeutics for managing inflammatory and oxidative stress-related conditions. 2. Material and methods 2.1 Collection and identification of plant material Tabernaemontana divaricata and Mangifera indica leaves were gathered from the surrounding area of Bhopal, Madhya Pradesh, and verified at RB Science, Bhopal. leaves were gathered from a nearby farm in Bhopal, Madhya Pradesh, and verified at RB Science, Bhopal. 2.2 Chemicals and reagents Analytical grade compounds were all that were used. Reagents are chemicals used to prepare buffers, analytical solutions, and other things for experiments. The leaves of Tabernaemontana divaricata and Mangifera indica were freshly collected from the local area, providing the primary plant materials for the study. Several chemicals were sourced from well-known suppliers: ethanol was procured from Loba Chemie (P) Ltd, Mumbai, while gallic acid and vanillin were obtained from CDH, New Delhi. Folin-Ciocalteu reagent was sourced from Avra, Hyderabad, and sodium carbonate and glacial acetic acid were acquired from Oxford Fine Chemicals, Mumbai. Methanol was provided by S.D. Fine Chemicals, Mumbai, and petroleum ether and chloroform were purchased from Rankem, Mumbai. Additionally, sulfuric acid was also supplied by Rankem, Mumbai. For experiments requiring water, freshly distilled water was prepared in the laboratory to ensure purity. 2.3 Extraction Process of leaves of Taebermontana divaricata and Mangifera indica Powdered leaves (91 g, Taebermontana divaricata ; 102 g Mangifera indica ) were separately sealed in the Soxhlet apparatus's extractor and defatted with petroleum ether utilizing heated continuous extraction process until colorless siphoning solution is obtained (5–6 h). The extraction After the solvent was eliminated, the marc was dried, and then extracted with a blend of ethanol-water (70:30) as the solvent. The extracts were filtered hot to remove impurities and the solvent was evaporated using rotary vacuum evaporator. To eliminate the oleo-resinous extract, it was gathered and put in a water bath remaining solvent and finally placed in desiccator maximum drying. The dried/semidried extracts were stored in desiccator for further experimental procedures. 2.4 Qualitative Phytochemical Screening Both thehydro-alcoholic extracts were assessed using phytochemical qualitative responses to determine whether typical plant secondary metabolites are present or absent. Many classes, including triterpenes/steroids, alkaloids, glycosides, flavonoids, saponins, tannins, and phenolics, were screened for. As analytical reactions to these tests, the color intensity or the precipitate formation were employed (Banu and Cathrine 2015). 2.4.1 Test for Alkaloids Mayer’s test : To a few ml of plant sample extract, two drops of Mayer’s reagent was added along the sides of test tube. Wagner's test : Test tube walls were lined with a few drops of Wagner's reagent mixed with a few milliliters of plant extract. Hager's test : In the test tube, a small amount of plant extract and a few drops of Hager's reagent are applied along the sides. Dragendroff’s Test : Each extract was mixed with one milliliter (ml) and a few drops of Dragendroff's solution. 2.4.2 Test for Glycosides Froth test : A test tube containing 1ml of the extract in water was filled and given a good shaking. Borntrager's test : The extract and 1.0 milliliter of diluted sulfuric acid were combined in a test tube and brought to a boil for five minutes. Following the chilling and shaking of the filtrate with an equivalent volume of dichloromethane, the bottom layer (dichloromethane) was separated and shaken with half its volume of diluted ammonia. Kedde’s test : After extracting the material using hloroform, evaporation dries it out. One drop of 90% alcohol and two drops of 2% 3, 5-dinitro benzoic acid (3, 5-dinitro benzene carboxylic acid, Kedde's reagent) in 90% alcohol should be added to the previously indicated residue. The solution becomes alkaline when 20% sodium hydroxide solution is added. Keller killiani test (Test for deoxy sugars) : Following a chloroform extraction, the extract is evaporated until it is completely dry. 0.4 milliliters of glacial acetic acid with a tiny amount of ferric chloride solution were added to the residual. After moving the mixture into a test tube, 0.5 ml of concentrated sulfuric acid was applied along the test tube's wall. 2.4.3 Test for Tannins and phenolic compounds Gelatin test : To the extract, a 1% gelatin solution containing 10% sodium chloride was added. Ferric chloride test : Iron chloride solution that had been newly made was added to the extract. Vanillin hydrochloride test : A few drops of vanillin hydrochloride reagent were added to the extract test solution for treatment. Alkaline reagent test : Sodium hydroxide solution was used to treat the extract test solution. 2.4.4 Test for flavonoids Shinoda test : To Conc. hydrochloric acid was added dropwise to the extract test solution along with a few pieces of magnesium ribbon. Zinc hydrochloride reduction test : A mixture of concentrated hydrochloric acid and zinc dust was added to the test solution. Alkaline reagent test : The test solution received a few drops of sodium hydroxide solution added to it. A small amount of strong hydrochloric acid was added. Later, if color showed. 2.4.5 Proteins and amino acids Millons test : The extract's Two milliliters of Millon's reagent (mercuric nitrate in nitric acid with traces of nitrous acid) were mixed with the test solution, and left to react. Ninhydrin test : A 0.2% ninhydrin solution was added to the extract solution and brought to a boil. 2.4.5 Sterols and terpenoids Libermann-Burchard test : After adding a few drops of acetic anhydride to the extract, it was heated and chilled. Concentrated sulfuric acid was poured into the test tube from the sides. Salkowski test : By dissolving the extract in chloroform, a little amount of concentrated sulfuric acid was included. After giving the mixture a good shake, it was allowed to stand for some time. 2.5 Chromatographic analysis of the extracts The extracts were dissolved in ethanol by sonication and filtered through a 0.45µ nylon syringe filter prior to injecting in the sample loop of HPLC instrument. Acetonitrile-buffer (pH 2.5) (15:85) was the solvent system used to extract Mangifera indica; the C18 column (chromosil, 5µ) was used to measure the flow rate, which was set at 1.0 mL per minute. The extract's mangiferin content was found using a 253 wavelength (KUMAR, SRIVASTAVA, and KUMAR 1 2008). After injecting a 20µL sample into the sample loop, the chromatogram was acquired. Using the aforementioned technique, a chromatogram of standard mangiferin (5µg/mL) was also produced. Acetonitrile-water (pH 2.5) (95:5) was the solvent system used to extract Taebermontana divaricata. A C8 column (chromosil, 5µ) was used to measure the flow rate, which was set at 1.5 mL per minute. The presence of ß-sitosterol in the extract was determined using a wavelength of 202 (Shah et al. 2010 ). An example 20µL was injected into the sample loop and the chromatogram was obtained. A chromatogram of standard ß-sitosterol (5µg/mL) was also chromatographed using the above method. 2.6 FT-IR analysis of the extracts The dried extracts were scanned in the range of 400 to 4000 cm-1 using a FT-IR spectrophotometer and the stretching and bending vibrations were observed. 2.7 Total Phenolic Content An adaptation of the procedure described served as the foundation for the extraction of phenolic chemicals. One dried extract (0.1 g) was dissolved in 5 mL of ethanol to determine the total phenolic content. The fixes served as the stock solutions for further studies and were kept in amber-colored bottles at 4°C (Mishra and Jain 2021). In order to ascertain the overall phenolic content One milliliter (200 µL) of the extract sample was combined with 1.4 milliliters Folin-Ciocalteu reagent (100 µL). After two minutes, add 300 µL of a 20% Na2CO3 aqueous solution, and let the mixture remain for two hours. A UV-Vis spectrophotometer was used to measure the absorbance at 765 nm. To construct the calibration curve, standard solutions containing 10–100 ppm of gallic acid were treated in a similar manner. With the same chemicals and 200 µL of methanol, the control solution was prepared and incubated similarly to the other samples. The results were expressed in milligrams per 100 mg of the dry material using the gallic acid equivalent (GAE). 2.8 Preparation the combined extracts for the antioxidant and anti-inflammatory effect The hydro-alcoholic extracts from Mangifera indica and Taebermontana divaricata were combined in three different ratios (1:1, 1:2, and 2:1), respectively, and the antioxidant and anti-inflammatory effect was assessed using the techniques described in the following sections. The statistical significance of the combined extracts' antioxidant and anti-inflammatory properties was examined by comparing them to those of the individual extracts. 2.9 Evaluation of Anti-oxidant Activity The steady free radical DPPH was used to gauge the test solution's free radical scavenging activity in terms of its capacity to donate hydrogen or scavenge radicals. The previously described approach was used to determine the DPPH radical scavenging activity (Amreen and Chaurey 2021). A 1 mM DPPH solution and an extract solution (100 µg/mL) were produced separately in ethanol. A 1.5 ml DPPH solution was mixed with 1.5 ml of the test solution. The equivalent blank solution, made with 3 mL of ethanol, was used to measure the absorbance at 517 nm. 3 mL of DPPH was the control sample that was used. Three duplicates of the assay were run. The following formula was used to determine the percentage inhibition of the free radical DPPH based on the control reading. $$\:\:\varvec{D}\varvec{P}\varvec{P}\varvec{H}\:\varvec{s}\varvec{c}\varvec{a}\varvec{v}\varvec{e}\varvec{n}\varvec{g}\varvec{e}\varvec{d}\:\left(\varvec{\%}\right)=\frac{Acom-Atest}{Acom}\times\:1000$$ Where, A con - is the absorbance of the control reaction, A test - is the absorbance in the presence of the test solution. 2.10 Animal Adult male Wistar rats weighing 250–300g were maintained in the Institute of Biological Science of the Federal University of Rio Grande at 22 ± 2ºC, with a relative humidity of 50–60% under a 12–12 h light-dark cycle with food and water ad libitum . The experiments were performed after approval of the protocol by the Institutional Ethics Committee (approval number P021/2013). 2.11 Experimental protocol Animals were distributed in six groups and received the following treatments: Group I - Control - treated with vehicle (normal saline) Group II– Taebermontana divaricata extract (200 mg/kg) Group III – Mangifera indica extract (200 mg/kg) Group IV – Combined extract 1:1 (100 mg/kg) Group V – Combined extract 1:2 (100 mg/kg) Group VI – Combined extract 2:1 (100 mg/kg) 2.12 Evaluation of anti-inflammatory action The anti-inflammatory activity of the extracts was assessed using the rat paw edema technique caused by carrageenan (Kemisetti and Manda 2018). To induce paw oedema, 0.1 mL (1% solution) of carrageenan was subcutaneously injected into the plantar surface of the rat's right hind paw. Thirty minutes prior to the carrageenan injection, each animal group received a dosage of extracts at a rate of 100 mg/kg. The following groups (n = 6) of animals were formed. Using a vernier caliper, the Paw diameters were measured immediately prior to the administration of carrageenan, and then at 1, 2, 4, and 6 hours later. The outcomes were contrasted with those of the control group. The following formula was used to determine each group's % suppression of paw inflammation: $$\:\varvec{\%}\:\varvec{i}\varvec{n}\varvec{h}\varvec{i}\varvec{b}\varvec{t}\varvec{i}\varvec{o}\varvec{n}\:=\:\frac{C-T}{C}\times\:100$$ Where, C = Paw volume (mL) in vehicle treated group (control), T = Paw volume (mL) in drug treated group 3. Results and Discussion 3.1 Extraction yield The defatting of the leaves was done by petroleum ether and the extraction of desired phytoconstituents by ethanol-water (70:30) as the solvent blend (Tables 1 and 2 ). Table 1 Extract yield and features of Mangifera indica Extraction solvent Marc weight Extract Weight Yield of extract Color of extract Petroleum Ether 102 g 1.4 g 1.37% Light Green Ethanol-water (70:30) 98.4 g 8.6 g 8.74% Dark Brown Table 2 Extract yield and features of Taebermonatana divaricata Extraction solvent Marc weight Extract Weight Yield of extract Color of extract Petroleum Ether 91 g 1.1 g 1.21% Light Brown Ethanol-water (70:30) 88.7 g 9.1 g 10.26% Dark Green It was found that the hydro-alcoholic solvent blend was able to extract more contents from T. divaricata as compared to M. indica while the fatty material in both the plants was almost similar. 3.2 Preliminary phytochemical Screening The extracts included alkaloids, flavonoids, triterpenoids, phenols, tannins, and saponins, as shown by the qualitative phytochemical screening. Table 3 Qualitative phytochemical screening observations. Chemical Tests Observation M. indica T. divaricata Test for Alkaloids Mayer’s reagent Cream colour precipitate + + Hager’s reagent Yellow colour precipitate - + Wagner’s reagent Reddish brown precipitate + + Dragendorff’s reagent Reddish brown precipitate + + Test for Glycosides Froth test Frothing is seen + + Kedde's Test No color - - Bontrager's Test Rose pink or red color in the ammonical layer not found - - Keller-Kiliani No color in acetic acid layer - - Test for Phenolics and Tannins Ferric chloride Blue green color - + Gelatin Solution White precipitate + + Alkaline reagent test Yellow to red precipitate + + Vanillin HCl test Purplish red color + + Test for Flavonoids Shinoda test Red color + + Alkaline reagent test Yellow color that turns red on acidification + + Zinc HCl reduction test Red color + + Test for Proteins Millon's Test White precipitate, turns red on heating - - Ninhydrin Test Voilet color - - Test for Sterols/triterpenoids Liberman-Burchard Test Brown ring at junction Upper layer turns green - + Salkowski Test Yellow color in lower layer - - 3.3 Chromatographic and spectral analysis of extracts The hydro-alcoholic extracts of both the plants were analyzed by HPLC for the presence of specific phytoconstituent (qualitatively). The M. indica extract was found to contain Mangiferin with retention time of 2.819 (Figs. 2 and 3 ), while the T. divaricata extract exhibited the presence of ß-sitosterol at retention time 5.82 minutes (Figs. 4 and 5 ). 3.4 FTIR results A broad peak around 3400 − 3200 cm⁻¹, which may correspond to O-H stretching vibrations, indicating the presence of hydroxyl groups (alcohols, phenols), 1700 cm⁻¹ suggests the presence of carbonyl groups, which could indicate the presence of ketones, aldehydes, or carboxylic acids, 2850–3000 cm⁻¹ suggest C-H stretching, which is typical in alkanes and other hydrocarbon chains, 1600 − 1500 cm⁻¹, which may indicate C = C stretching in aromatic rings, 1050–1250 cm⁻¹ is typical of C-O stretching vibrations, which can be found in alcohols, ethers, esters, and carboxylic acids (Fig. 6 ). The FTIR spectrum of Tabernaemontana divaricata extract presents distinct peaks indicative of various functional groups. A broad peak around 3400 − 3200 cm⁻¹ corresponds to O-H stretching, suggesting the presence of hydroxyl groups commonly found in alcohols and phenols. The peak near 1700 cm⁻¹ indicates C = O stretching, pointing to the presence of carbonyl groups, which may be associated with ketones, aldehydes, or carboxylic acids. Peaks around 2850–3000 cm⁻¹ represent C-H stretching vibrations typical of aliphatic hydrocarbons. The peak near 1600 cm⁻¹ is likely due to C = C stretching, often associated with aromatic rings. Additionally, the region around 1050–1250 cm⁻¹ suggests C-O stretching vibrations, indicating the presence of alcohols, ethers, or ester functional groups. This analysis highlights the diverse chemical nature of the T. divaricata extract, confirming the presence of multiple functional groups, including hydroxyl, carbonyl, and aromatic structures (Fig. 7 ). 3.5 Total Phenolic Content As one of the most stable and reasonably priced natural phenols, gallic acid was utilized as the benchmark for determining the total phenolic content. Using the blue color absorption data collected in the concentration range of 10 to 100 ppm, the standard curve for gallic acid was created (Table 4 and Fig. 8 ). Regression coefficient and Beer's law range were ascertained from this. Gallic acid's linearity equation was determined to be Abs = 0.006x − 0.0042. GAE mg/100 mg was the determined total Phenolic content of the samples Table 4 Calibration curve data for TPC. Concentration ppm Absorbance at 765 nm 0 0 10 0.051 20 0.118 30 0.173 40 0.233 50 0.291 60 0.348 70 0.422 80 0.475 90 0.550 100 0.598 The amount of phenolics present (total) in the M. indica and T. divaricata extracts were found to be 62.5 GAE mg/100 mg and GAE 55 mg/100 mg respectively. 3.6 Pharmacological evaluation The anti-oxidant and anti-inflammatory properties, extracts of M. indica and T. divaricata were combined in three different ratios. These combined extracts were labelled as C1, C2 and C3 containing 1:1, 1:2 and 2:1 ratio of M. indica and T. divaricata respectively. 3.6.1 DPPH radical scavenging assay Using the DPPH radical scavenging assay method, the combined extracts' antioxidant activity was ascertained. When appropriate reducing chemicals are used to react with DPPH radicals, the radicals lose color stoichiometric to the number of electrons consumed, which may be detected using a 517 nm using spectrophotometer. If DPPH has antioxidant properties, the deep purple color of the molecule becomes less intense. The antioxidant activity was found to be synergistically improved on combining the extracts (Table 5 and Fig. 9 ). Table 5 Inhibition of DPPH radical. Test solution C1 C2 C3 M. indica T. divaricata % DPPH inhibition 65.8 ± 0.361 82.9 ± 1.513 94.37 ± 0.702 44.8 ± 1.127 35.87 ± 1.464 It was found from the result that combining the extracts resulted in synergizing the antioxidant activity when compared to the individual extracts. The highest amount of DPPH inhibition was witnessed in C3 (94.37 ± 0.702%) where M. indica and T. divaricata were mixed in the ratio 2:1 (Fig. 9 ). 3.6.2 Anti-inflammatory activity The anti-inflammatory characteristics of the combined extracts were examined using the rat carrageenan-induced paw edema approach. Acute inflammation caused by carrageenan is one of the finest test methods for anti-inflammatory medication evaluation. Having the capacity to extracts and the mixed extracts to inhibit edema on pretreatment was calculated from the paw diameter obtained at different times post treatment (Table 6 and Fig. 10 ). Table 6 Rat paw diameter and edema inhibition. Group Paw thickness (mm)/Edema inhibition (%) 1h 2h 3h 4h Control 0.488 ± 0.032 0.674 ± 0.030 0.794 ± 0.017 0.832 ± 0.025 Ibuprofen 0.272 ± 0.036 (44.26) 0.352 ± 0.016 (47.77) 0.362 ± 0.041 (54.41) 0.216 ± 0.023 (74.04) M. indica 0.420 ± 0.056 (13.93) 0.446 ± 0.036 (33.83) 0.450 ± 0.037 (43.32) 0.414 ± 0.026 (50.24) T. divaricata 0.452 ± 0.024 (7.38) 0.508 ± 0.016 (24.63) 0.524 ± 0.009 (34.01) 0.442 ± 0.019 (46.88) C1 0.392 ± 0.020 (19.67) 0.434 ± 0.023 (35.61) 0.436 ± 0.019 (45.09) 0.402 ± 0.048 (51.68) C2 0.372 ± 0.027 (23.77) 0.422 ± 0.024 (37.39) 0.428 ± 0.016 (46.10) 0.376 ± 0.030 (54.81) C3 0.352 ± 0.029 (27.87) 0.406 ± 0.032 (39.76) 0.418 ± 0.013 (47.36) 0.326 ± 0.015 (60.82) 4. Conculsion The study concludes that the combination of Mangifera indica and Tabernaemontana divaricata leaf extracts significantly enhances their antioxidant and anti-inflammatory properties compared to the individual extracts. The observed synergistic effects, particularly in the 2:1 ratio of M. indica to T. divaricata , suggest that these plant extracts could be effectively utilized in developing natural therapeutics for managing conditions associated with oxidative stress and inflammation. The presence of bioactive compounds such as flavonoids and phenolics likely contributes to these enhanced activities, reinforcing the potential of these plant combinations in therapeutic applications. Research results led to the conclusion that combining the extracts greatly enhanced their anti-inflammatory and antioxidant properties. The greatest increase in activity was specifically observed when two parts M. indica extract and one part T. divaricata extract were used together. Declarations Author Contributions Ganesh Mangesh Walanj, Pranjali Thakare and Nasir Ahmad Sarwary: Conceptualization, methodology, Data collection, analysis, writing – original draft. Anand Kumar, Dilip KumarTiwari, Abhishek Sharma and Devesh Kumar: Data collection, project administration, writing – review & editing. Competing Interests The authors declare that there are no competing interests related to this publication. Ethics approval This study was conducted in accordance with the ethical standards of the Institutional Review Board of the College of Food Science and Technology, Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Ocean University, Zhanjiang, China, and the Laxmi Narayan College of Pharmacy, Department of Pharmacology, Rajiv Gandhi Proudyogiki Vishwavidyalaya, Bhopal, India. Informed consent was obtained from all participants involved in the study. All procedures performed were in line with the Declaration of Helsinki. Data Availability The datasets generated during and/or analyzed during the current study are available from the corresponding author upon reasonable request, especially for those in need of the data for further research. Consent to participate Informed consent was obtained from all individual participants included in the study. Informed consent was obtained from all participants prior to their inclusion in the study. Participants were fully informed about the nature, purpose, and potential risks of the research, and their participation was voluntary. Consent to publish All authors have given their consent for the publication of this manuscript. Additionally, informed consent was obtained from participants for the publication of any identifying information and data. References Amreen R, and Chaurey M (2021) Evaluation of Estrogenic Potential of Ethanolic and Aqueous Extract of Pitunia Hybrida. J Pharmacol Biomed 5(4):312–318. https://journals.indexcopernicus.com/api/file/viewByFileId/1388005 Banu SK, and Cathrine L (2015) General Techniques Involved in Phytochemical Analysis. Int J Adv Res Chem Sci 2(4):25–32. https://www.arcjournals.org/international-journal-of-advanced-research-in-chemical-science/volume-2-issue-4/5 Ediriweera MK, Tennekoon KH, and Samarakoon SR (2017) A Review on Ethnopharmacological Applications, Pharmacological Activities, and Bioactive Compounds of Mangifera Indica (Mango). Evidence-Based Complement Altern Med 2017(1):6949835. https://doi.org/10.1155/2017/6949835 Ghosh P, Poddar S, and Chatterjee S (2021) Morphological Features, Phytochemical and Ethnopharmacological Attributes of Tabernaemontana Divaricata Linn.: A Comprehensive Review. J Pharmacognosy Phytochemistry 10(6):31–36. https://doi.org/10.22271/phyto.2021.v10.i6a.14253 Kemisetti D, and Manda S (2018) Synthesis and Comparison of Peg-Ibuprofen and Peg-Ketoprofen Prodrugs by in Vitro and in Vivo Evaluation. J Drug Delivery Ther 8(4):145–154. https://doi.org/10.22270/jddt.v8i4.1743 KUMAR PRADEEP, SRIVASTAVA V, and KUMAR L (2008) Development of New Reversed-Phase HPLC Method for the Determination of Mangiferin in Mangifera Indica Linn. Biosci Biotechnol Res Asia 5(1):383–388. https://www.biotech-asia.org/vol5no1/development-of-new-reversed-phase-hplc-method-for-the-determination-of-mangiferin-in-mangifera-indica-linn/ Leyane TS, Jere SW, Houreld NN (2022) Oxidative Stress in Ageing and Chronic Degenerative Pathologies: Molecular Mechanisms Involved in Counteracting Oxidative Stress and Chronic Inflammation. Int J Mol Sci 23(13):7273. https://doi.org/10.3390/ijms23137273 Mirza B et al (2021) Mango (Mangifera Indica L.): A Magnificent Plant with Cancer Preventive and Anticancer Therapeutic Potential. Crit Rev Food Sci Nutr 61(13):2125–2151. https://doi.org/10.1080/10408398.2020.1771678 Mishra S, and Jain S (2021) Hepatoprotective and Antioxidant Effect of Delonix Regia Leaf Extract against D Galactosamine Induced Oxidative Stress in Rats. J Pharmacol Biomed 5(3):326. https://journals.indexcopernicus.com/api/file/viewByFileId/1388010 Pandey M, Debnath M, Gupta S, Chikara SK (2011) Phytomedicine: An Ancient Approach Turning into Future Potential Source of Therapeutics. J Pharmacognosy phytotherapy 3(3):27–37. https://www.researchgate.net/publication/215511510_Phytomedicine_An_ancient_approach_turning_into_future_potential_source_of_therapeutics Shah UM et al (2010) Development and Validation of a Simple Isocratic HPLC Method for Simultaneous Estimation of Phytosterols in Cissus Quadrangularis. Indian J Pharm Sci 72(6):753. https://doi.org/10.4103/0250-474x.84587 Additional Declarations The authors declare no competing interests. Supplementary Files paper1Tabernaemontanna.docx 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-5344453","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":371057174,"identity":"cb1e04cd-3f3d-46f2-870c-2aaecf2bccdf","order_by":0,"name":"Ganesh Mangesh Walanj","email":"","orcid":"","institution":"1.\tLaxmi Narayan College of Pharmacy Bhopal, Department of Pharmacology, Rajiv Gandhi Proudyogiki Vishwavidyalaya Bhopal, India","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ganesh","middleName":"Mangesh","lastName":"Walanj","suffix":""},{"id":371057928,"identity":"8201f431-7e2c-4b70-b2dc-9b03122704dd","order_by":1,"name":"Anand Kumar","email":"","orcid":"","institution":"2.\tCollege of Food Science and Technology, Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Ocean University, Zhanjiang china","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Anand","middleName":"","lastName":"Kumar","suffix":""},{"id":371057929,"identity":"88cbe761-2446-4e7e-9978-49274df82652","order_by":2,"name":"Dilip KumarTiwari","email":"","orcid":"","institution":"3.\tLaxmi Narayan College of Pharmacy Bhopal, Department of Pharmacology, Rajiv Gandhi Proudyogiki Vishwavidyalaya Bhopal, India","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Dilip","middleName":"","lastName":"KumarTiwari","suffix":""},{"id":371057930,"identity":"75088f35-c6ea-4b2d-840d-eeb2a54d0841","order_by":3,"name":"Abhishek Sharma","email":"","orcid":"","institution":"4.\tLaxmi Narayan College of Pharmacy Bhopal, Department of Pharmacology, Rajiv Gandhi Proudyogiki Vishwavidyalaya Bhopal, India","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Abhishek","middleName":"","lastName":"Sharma","suffix":""},{"id":371057931,"identity":"96aa534d-0f31-496e-af3c-84a62d803372","order_by":4,"name":"Pranjali Thakare","email":"","orcid":"","institution":"5.\tLaxmi Narayan College of Pharmacy Bhopal, Department of Pharmacology, Rajiv Gandhi Proudyogiki Vishwavidyalaya Bhopal, India","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Pranjali","middleName":"","lastName":"Thakare","suffix":""},{"id":371057932,"identity":"371f0b1a-7c1e-42b0-a515-cd82cfa4c9bd","order_by":5,"name":"Devesh Kumar","email":"","orcid":"","institution":"6.\tInstitute of Pharmaceutical Research, GLA University, Mathura- 281406, Uttar Pradesh, India","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Devesh","middleName":"","lastName":"Kumar","suffix":""},{"id":371057933,"identity":"6e8760bc-b8df-487c-ae6c-b4526f1fedb7","order_by":6,"name":"Nasir Ahmad Sarwary","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABEUlEQVRIie2PsWrDMBRFbVw06QNs+hMvFEQDxv6QLgoGeUnpWmig8pKOXVPoLxQ6dZYx1ItTrYIsMQV3yRDvplTxULzIzViIzvD0hO7hIsexWP4hIBxH/N7a71BPNxOGdM/lUHFXiB0OPqqEw1cPo6JfxpWyrIvbLrqBct2cYyyjl4dCtyzCK5NCqjnk62Uyfa1SEjz7m+StmmnlnV1zkyLmTp5xD4hgyN/BJiFCKy4vjMqF/NrmvLsHIhvkY/qREFmPKxNFIeeoAKIYCrAQEVF/tEzUDvJsWUKsGm/6xBNKlG6hI38BmX62vLuD4JG5quVRTGRab/eL0KgMOfP1mPVJekT8gLfXIz4ybLFYLCfED6onbf4AylbDAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0009-0009-2569-7274","institution":"7.\tKabul University, Department of Food technology and Hygiene, Faculty of Veterinary Sciences, Kabul, Afghanistan","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Nasir","middleName":"Ahmad","lastName":"Sarwary","suffix":""}],"badges":[],"createdAt":"2024-10-28 06:24:32","currentVersionCode":1,"declarations":{"humanSubjects":true,"vertebrateSubjects":true,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":true,"humanSubjectConsent":true,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":true,"coiExplicitlySet":false},"doi":"10.21203/rs.3.rs-5344453/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5344453/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":67737334,"identity":"183ff468-701b-4dab-8d01-a269e14c48b3","added_by":"auto","created_at":"2024-10-29 08:14:45","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":147483,"visible":true,"origin":"","legend":"\u003cp\u003eExtraction yield of \u003cem\u003eMangifera indica\u003c/em\u003e and \u003cem\u003eTaebermontana divaricata\u003c/em\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5344453/v1/2a9e9bb86f683ff9f673d45f.png"},{"id":67737327,"identity":"f9278efa-e0ce-4133-b728-345b4607d828","added_by":"auto","created_at":"2024-10-29 08:14:45","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":75339,"visible":true,"origin":"","legend":"\u003cp\u003eHPLC of \u003cem\u003eM. indica\u003c/em\u003e extract.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5344453/v1/b3449e95d9d31cf47d12fd8a.png"},{"id":67737330,"identity":"a9cf4767-24a8-4db5-a529-66820e6c1ad7","added_by":"auto","created_at":"2024-10-29 08:14:45","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":52460,"visible":true,"origin":"","legend":"\u003cp\u003eHPLC chromatogram of mangiferin.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5344453/v1/8fdf38a6777deb9d7174c9fc.png"},{"id":67737731,"identity":"3132d54a-5ae6-4191-9605-1d0631cfe37c","added_by":"auto","created_at":"2024-10-29 08:22:45","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":77053,"visible":true,"origin":"","legend":"\u003cp\u003eHPLC of \u003cem\u003eT. divaricata\u003c/em\u003e extract.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5344453/v1/2dad8a83f3c87ee533ac1007.png"},{"id":67737333,"identity":"954d35a0-a98f-4f25-a244-53d3d93261e0","added_by":"auto","created_at":"2024-10-29 08:14:45","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":33026,"visible":true,"origin":"","legend":"\u003cp\u003eHPLC chromatogram of ß-sitosterol.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-5344453/v1/509a82d64827a6f88ff13b9e.png"},{"id":67737331,"identity":"a05dd8e7-4db5-469c-8f0c-e293f835b3da","added_by":"auto","created_at":"2024-10-29 08:14:45","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":52185,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR spectra of \u003cem\u003eM. indica\u003c/em\u003e extract.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-5344453/v1/06b9fd18c3ec73d573c84d19.png"},{"id":67737733,"identity":"8a2734f5-3ebc-4e38-91c9-7351191f0247","added_by":"auto","created_at":"2024-10-29 08:22:45","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":51729,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR spectra of \u003cem\u003eT. divaricata\u003c/em\u003e extract.\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-5344453/v1/007f91e96611546fe69f5861.png"},{"id":67737335,"identity":"c783ebd9-82ec-4f02-9f99-4544dee6c280","added_by":"auto","created_at":"2024-10-29 08:14:45","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":64877,"visible":true,"origin":"","legend":"\u003cp\u003eCalibration curve of gallic acid.\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-5344453/v1/710400b6b89e48bb5ab4aaa3.png"},{"id":67737328,"identity":"d7395593-84c3-443f-ae6e-cdd728d8003b","added_by":"auto","created_at":"2024-10-29 08:14:45","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":24242,"visible":true,"origin":"","legend":"\u003cp\u003ePercentage (%) DPPH inhibition by mixed and individual extracts.\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-5344453/v1/860d0e2984d89b31b2e61d5a.png"},{"id":67737336,"identity":"c55d2800-1ded-4414-bc58-1a0ba1fd2d6f","added_by":"auto","created_at":"2024-10-29 08:14:45","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":165154,"visible":true,"origin":"","legend":"\u003cp\u003ePercentage (%) Edema inhibition.\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-5344453/v1/4755f15fcafbc12995149296.png"},{"id":67740018,"identity":"edc146dc-584c-4db6-879a-d21366b69db1","added_by":"auto","created_at":"2024-10-29 08:38:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1659560,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5344453/v1/e1a7c6cd-70a3-4c62-aa15-e79703458d7c.pdf"},{"id":67739004,"identity":"59f54496-a614-4d26-b227-50178da82148","added_by":"auto","created_at":"2024-10-29 08:30:45","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":556356,"visible":true,"origin":"","legend":"","description":"","filename":"paper1Tabernaemontanna.docx","url":"https://assets-eu.researchsquare.com/files/rs-5344453/v1/80de95a5e9613b162e6a8792.docx"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003e\u003cstrong\u003eIn Vitro and In-Vivo Analysis of the Combined Extract of Tabernaemontana divaricata and Mangifera indica Leaves for Enhanced Anti-Inflammatory and Antioxidant Properties\u003c/strong\u003e\u003c/p\u003e","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eInflammation and oxidative stress are two pivotal processes implicated in the pathogenesis of various chronic diseases, including cardiovascular disorders, cancer, diabetes, and neurodegenerative conditions (Leyane, Jere, and Houreld \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The increasing prevalence of these conditions has heightened the search for natural remedies that can offer effective therapeutic interventions with minimal side effects. In this context, plants and their bioactive compounds have garnered significant attention due to their historical use in traditional medicine and their potential in modern therapeutics (Pandey et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eTabernaemontana divaricata\u003c/em\u003e, commonly known as the crepe jasmine (Ghosh, Poddar, and Chatterjee 2021). \u003cem\u003eTabernaemontana divaricata\u003c/em\u003e belongs to the family Apocynaceae. This family is commonly known as the dogbane family, and it includes a wide variety of flowering plants, many of which have medicinal properties. \u003cem\u003eTabernaemontana divaricata\u003c/em\u003e is widely used in Ayurvedic medicine, primarily for its anti-inflammatory, analgesic, and wound-healing properties. The plant is rich in alkaloids, flavonoids, and terpenoids, which contribute to its therapeutic effects.\u003c/p\u003e \u003cp\u003e \u003cem\u003eMangifera indica\u003c/em\u003e, known as the mango tree (Ghosh, Poddar, and Chatterjee 2021) is the plants traditionally recognized for their medicinal properties. \u003cem\u003eMangifera indica\u003c/em\u003e belongs to the family Anacardiaceae, which is known for including several economically important fruit-bearing trees and plants. \u003cem\u003eMangifera indica\u003c/em\u003e, on the other hand, is renowned not only for its nutritional value but also for its extensive medicinal applications (Ediriweera, Tennekoon, and Samarakoon 2017), particularly in treating inflammation, infections, and oxidative stress-related conditions. The leaves of \u003cem\u003eMangifera indica\u003c/em\u003e are known to contain polyphenols, flavonoids, and other potent antioxidants (Mirza et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eGiven the individual therapeutic properties of these plants, combining the extracts of \u003cem\u003eTabernaemontana divaricata\u003c/em\u003e and \u003cem\u003eMangifera indica\u003c/em\u003e leaves presents a promising approach to enhancing their anti-inflammatory and antioxidant effects. The rationale behind this combination is to leverage the synergistic effects of the bioactive compounds present in both plants, which may result in a more potent therapeutic outcome than the individual extracts.\u003c/p\u003e \u003cp\u003eThis study aims to explore the anti-inflammatory and antioxidant properties of the combined extract of \u003cem\u003eTabernaemontana divaricata\u003c/em\u003e and \u003cem\u003eMangifera indica\u003c/em\u003e leaves. By evaluating these effects through various in vitro and in vivo models, the research seeks to establish a scientific basis for the traditional use of these plants and potentially contribute to the development of new natural therapeutics for managing inflammatory and oxidative stress-related conditions.\u003c/p\u003e"},{"header":"2. Material and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Collection and identification of plant material\u003c/h2\u003e \u003cp\u003e \u003cem\u003eTabernaemontana divaricata\u003c/em\u003e and \u003cem\u003eMangifera indica\u003c/em\u003e leaves were gathered from the surrounding area of Bhopal, Madhya Pradesh, and verified at RB Science, Bhopal. leaves were gathered from a nearby farm in Bhopal, Madhya Pradesh, and verified at RB Science, Bhopal.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Chemicals and reagents\u003c/h2\u003e \u003cp\u003eAnalytical grade compounds were all that were used. Reagents are chemicals used to prepare buffers, analytical solutions, and other things for experiments. The leaves of \u003cem\u003eTabernaemontana divaricata\u003c/em\u003e and \u003cem\u003eMangifera indica\u003c/em\u003e were freshly collected from the local area, providing the primary plant materials for the study. Several chemicals were sourced from well-known suppliers: ethanol was procured from Loba Chemie (P) Ltd, Mumbai, while gallic acid and vanillin were obtained from CDH, New Delhi. Folin-Ciocalteu reagent was sourced from Avra, Hyderabad, and sodium carbonate and glacial acetic acid were acquired from Oxford Fine Chemicals, Mumbai. Methanol was provided by S.D. Fine Chemicals, Mumbai, and petroleum ether and chloroform were purchased from Rankem, Mumbai. Additionally, sulfuric acid was also supplied by Rankem, Mumbai. For experiments requiring water, freshly distilled water was prepared in the laboratory to ensure purity.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Extraction Process of leaves of \u003cem\u003eTaebermontana divaricata\u003c/em\u003e and \u003cem\u003eMangifera indica\u003c/em\u003e\u003c/h2\u003e \u003cp\u003ePowdered leaves (91 g, \u003cem\u003eTaebermontana divaricata\u003c/em\u003e; 102 g \u003cem\u003eMangifera indica\u003c/em\u003e) were separately sealed in the Soxhlet apparatus's extractor and defatted with petroleum ether utilizing heated continuous extraction process until colorless siphoning solution is obtained (5\u0026ndash;6 h). The extraction After the solvent was eliminated, the marc was dried, and then extracted with a blend of ethanol-water (70:30) as the solvent. The extracts were filtered hot to remove impurities and the solvent was evaporated using rotary vacuum evaporator. To eliminate the oleo-resinous extract, it was gathered and put in a water bath remaining solvent and finally placed in desiccator maximum drying. The dried/semidried extracts were stored in desiccator for further experimental procedures.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Qualitative Phytochemical Screening\u003c/h2\u003e \u003cp\u003eBoth thehydro-alcoholic extracts were assessed using phytochemical qualitative responses to determine whether typical plant secondary metabolites are present or absent. Many classes, including triterpenes/steroids, alkaloids, glycosides, flavonoids, saponins, tannins, and phenolics, were screened for. As analytical reactions to these tests, the color intensity or the precipitate formation were employed (Banu and Cathrine 2015).\u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section3\"\u003e \u003ch2\u003e\u003cb\u003e2.4.1 Test for Alkaloids\u003c/b\u003e\u003c/h2\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eMayer\u0026rsquo;s test\u003c/b\u003e: To a few ml of plant sample extract, two drops of Mayer\u0026rsquo;s reagent was added along the sides of test tube.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eWagner's test\u003c/b\u003e: Test tube walls were lined with a few drops of Wagner's reagent mixed with a few milliliters of plant extract.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eHager's test\u003c/b\u003e: In the test tube, a small amount of plant extract and a few drops of Hager's reagent are applied along the sides.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eDragendroff\u0026rsquo;s Test\u003c/b\u003e: Each extract was mixed with one milliliter (ml) and a few drops of Dragendroff's solution.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e2.4.2 Test for Glycosides\u003c/h2\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eFroth test\u003c/b\u003e: A test tube containing 1ml of the extract in water was filled and given a good shaking.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eBorntrager's test\u003c/b\u003e: The extract and 1.0 milliliter of diluted sulfuric acid were combined in a test tube and brought to a boil for five minutes. Following the chilling and shaking of the filtrate with an equivalent volume of dichloromethane, the bottom layer (dichloromethane) was separated and shaken with half its volume of diluted ammonia.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eKedde\u0026rsquo;s test\u003c/b\u003e: After extracting the material using hloroform, evaporation dries it out. One drop of 90% alcohol and two drops of 2% 3, 5-dinitro benzoic acid (3, 5-dinitro benzene carboxylic acid, Kedde's reagent) in 90% alcohol should be added to the previously indicated residue. The solution becomes alkaline when 20% sodium hydroxide solution is added.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eKeller killiani test (Test for deoxy sugars)\u003c/b\u003e: Following a chloroform extraction, the extract is evaporated until it is completely dry. 0.4 milliliters of glacial acetic acid with a tiny amount of ferric chloride solution were added to the residual. After moving the mixture into a test tube, 0.5 ml of concentrated sulfuric acid was applied along the test tube's wall.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003e\u003cb\u003e2.4.3 Test for Tannins and phenolic compounds\u003c/b\u003e\u003c/h2\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eGelatin test\u003c/b\u003e: To the extract, a 1% gelatin solution containing 10% sodium chloride was added.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eFerric chloride test\u003c/b\u003e: Iron chloride solution that had been newly made was added to the extract.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eVanillin hydrochloride test\u003c/b\u003e: A few drops of vanillin hydrochloride reagent were added to the extract test solution for treatment.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eAlkaline reagent test\u003c/b\u003e: Sodium hydroxide solution was used to treat the extract test solution.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003e2.4.4 Test for flavonoids\u003c/h2\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eShinoda test\u003c/b\u003e: To Conc. hydrochloric acid was added dropwise to the extract test solution along with a few pieces of magnesium ribbon.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eZinc hydrochloride reduction test\u003c/b\u003e: A mixture of concentrated hydrochloric acid and zinc dust was added to the test solution.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eAlkaline reagent test\u003c/b\u003e: The test solution received a few drops of sodium hydroxide solution added to it. A small amount of strong hydrochloric acid was added. Later, if color showed.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section3\"\u003e \u003ch2\u003e2.4.5 Proteins and amino acids\u003c/h2\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eMillons test\u003c/b\u003e: The extract's Two milliliters of Millon's reagent (mercuric nitrate in nitric acid with traces of nitrous acid) were mixed with the test solution, and left to react.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eNinhydrin test\u003c/b\u003e: A 0.2% ninhydrin solution was added to the extract solution and brought to a boil.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e \u003ch2\u003e2.4.5 Sterols and terpenoids\u003c/h2\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eLibermann-Burchard test\u003c/b\u003e: After adding a few drops of acetic anhydride to the extract, it was heated and chilled. Concentrated sulfuric acid was poured into the test tube from the sides.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eSalkowski test\u003c/b\u003e: By dissolving the extract in chloroform, a little amount of concentrated sulfuric acid was included. After giving the mixture a good shake, it was allowed to stand for some time.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Chromatographic analysis of the extracts\u003c/h2\u003e \u003cp\u003eThe extracts were dissolved in ethanol by sonication and filtered through a 0.45\u0026micro; nylon syringe filter prior to injecting in the sample loop of HPLC instrument.\u003c/p\u003e \u003cp\u003eAcetonitrile-buffer (pH 2.5) (15:85) was the solvent system used to extract Mangifera indica; the C18 column (chromosil, 5\u0026micro;) was used to measure the flow rate, which was set at 1.0 mL per minute. The extract's mangiferin content was found using a 253 wavelength (KUMAR, SRIVASTAVA, and KUMAR\u003csup\u003e1\u003c/sup\u003e 2008). After injecting a 20\u0026micro;L sample into the sample loop, the chromatogram was acquired. Using the aforementioned technique, a chromatogram of standard mangiferin (5\u0026micro;g/mL) was also produced. Acetonitrile-water (pH 2.5) (95:5) was the solvent system used to extract Taebermontana divaricata. A C8 column (chromosil, 5\u0026micro;) was used to measure the flow rate, which was set at 1.5 mL per minute. The presence of \u0026szlig;-sitosterol in the extract was determined using a wavelength of 202 (Shah et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). An example 20\u0026micro;L was injected into the sample loop and the chromatogram was obtained. A chromatogram of standard \u0026szlig;-sitosterol (5\u0026micro;g/mL) was also chromatographed using the above method.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e2.6 FT-IR analysis of the extracts\u003c/h2\u003e \u003cp\u003eThe dried extracts were scanned in the range of 400 to 4000 cm-1 using a FT-IR spectrophotometer and the stretching and bending vibrations were observed.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e2.7 Total Phenolic Content\u003c/h2\u003e \u003cp\u003eAn adaptation of the procedure described served as the foundation for the extraction of phenolic chemicals. One dried extract (0.1 g) was dissolved in 5 mL of ethanol to determine the total phenolic content. The fixes served as the stock solutions for further studies and were kept in amber-colored bottles at 4\u0026deg;C (Mishra and Jain 2021).\u003c/p\u003e \u003cp\u003eIn order to ascertain the overall phenolic content One milliliter (200 \u0026micro;L) of the extract sample was combined with 1.4 milliliters Folin-Ciocalteu reagent (100 \u0026micro;L). After two minutes, add 300 \u0026micro;L of a 20% Na2CO3 aqueous solution, and let the mixture remain for two hours. A UV-Vis spectrophotometer was used to measure the absorbance at 765 nm. To construct the calibration curve, standard solutions containing 10\u0026ndash;100 ppm of gallic acid were treated in a similar manner. With the same chemicals and 200 \u0026micro;L of methanol, the control solution was prepared and incubated similarly to the other samples. The results were expressed in milligrams per 100 mg of the dry material using the gallic acid equivalent (GAE).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003e2.8 Preparation the combined extracts for the antioxidant and anti-inflammatory effect\u003c/h2\u003e \u003cp\u003eThe hydro-alcoholic extracts from \u003cem\u003eMangifera indica\u003c/em\u003e and \u003cem\u003eTaebermontana divaricata\u003c/em\u003e were combined in three different ratios (1:1, 1:2, and 2:1), respectively, and the antioxidant and anti-inflammatory effect was assessed using the techniques described in the following sections. The statistical significance of the combined extracts' antioxidant and anti-inflammatory properties was examined by comparing them to those of the individual extracts.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003e2.9 Evaluation of Anti-oxidant Activity\u003c/h2\u003e \u003cp\u003eThe steady free radical DPPH was used to gauge the test solution's free radical scavenging activity in terms of its capacity to donate hydrogen or scavenge radicals. The previously described approach was used to determine the DPPH radical scavenging activity (Amreen and Chaurey 2021). A 1 mM DPPH solution and an extract solution (100 \u0026micro;g/mL) were produced separately in ethanol. A 1.5 ml DPPH solution was mixed with 1.5 ml of the test solution. The equivalent blank solution, made with 3 mL of ethanol, was used to measure the absorbance at 517 nm. 3 mL of DPPH was the control sample that was used. Three duplicates of the assay were run. The following formula was used to determine the percentage inhibition of the free radical DPPH based on the control reading.\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\:\\:\\varvec{D}\\varvec{P}\\varvec{P}\\varvec{H}\\:\\varvec{s}\\varvec{c}\\varvec{a}\\varvec{v}\\varvec{e}\\varvec{n}\\varvec{g}\\varvec{e}\\varvec{d}\\:\\left(\\varvec{\\%}\\right)=\\frac{Acom-Atest}{Acom}\\times\\:1000$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eWhere, A \u003csub\u003econ\u003c/sub\u003e - is the absorbance of the control reaction, A \u003csub\u003etest\u003c/sub\u003e - is the absorbance in the presence of the test solution.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003e2.10 Animal\u003c/h2\u003e \u003cp\u003eAdult male Wistar rats weighing 250\u0026ndash;300g were maintained in the Institute of Biological Science of the Federal University of Rio Grande at 22\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026ordm;C, with a relative humidity of 50\u0026ndash;60% under a 12\u0026ndash;12 h light-dark cycle with food and water \u003cem\u003ead libitum\u003c/em\u003e. The experiments were performed after approval of the protocol by the Institutional Ethics Committee (approval number P021/2013).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003e2.11 Experimental protocol\u003c/h2\u003e \u003cp\u003eAnimals were distributed in six groups and received the following treatments:\u003c/p\u003e \u003cp\u003eGroup I - Control - treated with vehicle (normal saline)\u003c/p\u003e \u003cp\u003eGroup II\u0026ndash; \u003cem\u003eTaebermontana divaricata\u003c/em\u003e extract (200 mg/kg)\u003c/p\u003e \u003cp\u003eGroup III \u0026ndash; \u003cem\u003eMangifera indica\u003c/em\u003e extract (200 mg/kg)\u003c/p\u003e \u003cp\u003eGroup IV \u0026ndash; Combined extract 1:1 (100 mg/kg)\u003c/p\u003e \u003cp\u003eGroup V \u0026ndash; Combined extract 1:2 (100 mg/kg)\u003c/p\u003e \u003cp\u003eGroup VI \u0026ndash; Combined extract 2:1 (100 mg/kg)\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003e2.12 Evaluation of anti-inflammatory action\u003c/h2\u003e \u003cp\u003eThe anti-inflammatory activity of the extracts was assessed using the rat paw edema technique caused by carrageenan (Kemisetti and Manda 2018).\u003c/p\u003e \u003cp\u003eTo induce paw oedema, 0.1 mL (1% solution) of carrageenan was subcutaneously injected into the plantar surface of the rat's right hind paw. Thirty minutes prior to the carrageenan injection, each animal group received a dosage of extracts at a rate of 100 mg/kg. The following groups (n\u0026thinsp;=\u0026thinsp;6) of animals were formed.\u003c/p\u003e \u003cp\u003eUsing a vernier caliper, the Paw diameters were measured immediately prior to the administration of carrageenan, and then at 1, 2, 4, and 6 hours later. The outcomes were contrasted with those of the control group. The following formula was used to determine each group's % suppression of paw inflammation:\u003cdiv id=\"Equb\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equb\" name=\"EquationSource\"\u003e\n$$\\:\\varvec{\\%}\\:\\varvec{i}\\varvec{n}\\varvec{h}\\varvec{i}\\varvec{b}\\varvec{t}\\varvec{i}\\varvec{o}\\varvec{n}\\:=\\:\\frac{C-T}{C}\\times\\:100$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eWhere, C\u0026thinsp;=\u0026thinsp;Paw volume (mL) in vehicle treated group (control), T\u0026thinsp;=\u0026thinsp;Paw volume (mL) in drug treated group\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results and Discussion","content":"\u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003e3.1 Extraction yield\u003c/h2\u003e \u003cp\u003eThe defatting of the leaves was done by petroleum ether and the extraction of desired phytoconstituents by ethanol-water (70:30) as the solvent blend (Tables\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eExtract yield and features of \u003cem\u003eMangifera indica\u003c/em\u003e\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=\"char\" char=\".\" 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\u003eExtraction solvent\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMarc weight\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eExtract Weight\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eYield of extract\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eColor of extract\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePetroleum Ether\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e102 g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.4 g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.37%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eLight Green\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEthanol-water (70:30)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e98.4 g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.6 g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8.74%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eDark Brown\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \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\u003eExtract yield and features of \u003cem\u003eTaebermonatana divaricata\u003c/em\u003e\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=\"char\" char=\".\" 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\u003eExtraction solvent\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMarc weight\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eExtract Weight\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eYield of extract\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eColor of extract\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePetroleum Ether\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e91 g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.1 g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.21%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eLight Brown\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEthanol-water (70:30)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e88.7 g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e9.1 g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10.26%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eDark Green\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIt was found that the hydro-alcoholic solvent blend was able to extract more contents from \u003cem\u003eT. divaricata\u003c/em\u003e as compared to \u003cem\u003eM. indica\u003c/em\u003e while the fatty material in both the plants was almost similar.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec23\" class=\"Section2\"\u003e \u003ch2\u003e3.2 Preliminary phytochemical Screening\u003c/h2\u003e \u003cp\u003eThe extracts included alkaloids, flavonoids, triterpenoids, phenols, tannins, and saponins, as shown by the qualitative phytochemical screening.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eQualitative phytochemical screening observations.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChemical Tests\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eObservation\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eM. indica\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eT. divaricata\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e \u003cp\u003eTest for Alkaloids\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMayer\u0026rsquo;s reagent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCream colour precipitate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHager\u0026rsquo;s reagent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eYellow colour precipitate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWagner\u0026rsquo;s reagent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eReddish brown precipitate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDragendorff\u0026rsquo;s reagent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eReddish brown precipitate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTest for Glycosides\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFroth test\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFrothing is seen\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eKedde's Test\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo color\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003e-\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e-\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBontrager's Test\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRose pink or red color in the ammonical layer not found\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eKeller-Kiliani\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo color in acetic acid layer\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003e-\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e-\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTest for Phenolics and Tannins\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFerric chloride\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBlue green color\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGelatin Solution\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eWhite precipitate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAlkaline reagent test\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eYellow to red precipitate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVanillin HCl test\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePurplish red color\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTest for Flavonoids\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eShinoda test\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRed color\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAlkaline reagent test\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eYellow color that turns red on acidification\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eZinc HCl reduction test\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRed color\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTest for Proteins\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMillon's Test\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eWhite precipitate, turns red on heating\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNinhydrin Test\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eVoilet color\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTest for Sterols/triterpenoids\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLiberman-Burchard Test\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBrown ring at junction Upper layer turns green\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSalkowski Test\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eYellow color in lower layer\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003e3.3 Chromatographic and spectral analysis of extracts\u003c/h2\u003e \u003cp\u003eThe hydro-alcoholic extracts of both the plants were analyzed by HPLC for the presence of specific phytoconstituent (qualitatively). The \u003cem\u003eM. indica\u003c/em\u003e extract was found to contain Mangiferin with retention time of 2.819 (Figs.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e), while the \u003cem\u003eT. divaricata\u003c/em\u003e extract exhibited the presence of \u0026szlig;-sitosterol at retention time 5.82 minutes (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e and \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec25\" class=\"Section2\"\u003e \u003ch2\u003e3.4 FTIR results\u003c/h2\u003e \u003cp\u003eA broad peak around 3400\u0026thinsp;\u0026minus;\u0026thinsp;3200 cm⁻\u0026sup1;, which may correspond to O-H stretching vibrations, indicating the presence of hydroxyl groups (alcohols, phenols), 1700 cm⁻\u0026sup1; suggests the presence of carbonyl groups, which could indicate the presence of ketones, aldehydes, or carboxylic acids, 2850\u0026ndash;3000 cm⁻\u0026sup1; suggest C-H stretching, which is typical in alkanes and other hydrocarbon chains, 1600\u0026thinsp;\u0026minus;\u0026thinsp;1500 cm⁻\u0026sup1;, which may indicate C\u0026thinsp;=\u0026thinsp;C stretching in aromatic rings, 1050\u0026ndash;1250 cm⁻\u0026sup1; is typical of C-O stretching vibrations, which can be found in alcohols, ethers, esters, and carboxylic acids (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe FTIR spectrum of \u003cem\u003eTabernaemontana divaricata\u003c/em\u003e extract presents distinct peaks indicative of various functional groups. A broad peak around 3400\u0026thinsp;\u0026minus;\u0026thinsp;3200 cm⁻\u0026sup1; corresponds to O-H stretching, suggesting the presence of hydroxyl groups commonly found in alcohols and phenols. The peak near 1700 cm⁻\u0026sup1; indicates C\u0026thinsp;=\u0026thinsp;O stretching, pointing to the presence of carbonyl groups, which may be associated with ketones, aldehydes, or carboxylic acids. Peaks around 2850\u0026ndash;3000 cm⁻\u0026sup1; represent C-H stretching vibrations typical of aliphatic hydrocarbons. The peak near 1600 cm⁻\u0026sup1; is likely due to C\u0026thinsp;=\u0026thinsp;C stretching, often associated with aromatic rings. Additionally, the region around 1050\u0026ndash;1250 cm⁻\u0026sup1; suggests C-O stretching vibrations, indicating the presence of alcohols, ethers, or ester functional groups. This analysis highlights the diverse chemical nature of the \u003cem\u003eT. divaricata\u003c/em\u003e extract, confirming the presence of multiple functional groups, including hydroxyl, carbonyl, and aromatic structures (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec26\" class=\"Section2\"\u003e \u003ch2\u003e3.5 Total Phenolic Content\u003c/h2\u003e \u003cp\u003eAs one of the most stable and reasonably priced natural phenols, gallic acid was utilized as the benchmark for determining the total phenolic content. Using the blue color absorption data collected in the concentration range of 10 to 100 ppm, the standard curve for gallic acid was created (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e). Regression coefficient and Beer's law range were ascertained from this. Gallic acid's linearity equation was determined to be Abs\u0026thinsp;=\u0026thinsp;0.006x \u0026minus;\u0026thinsp;0.0042. GAE mg/100 mg was the determined total Phenolic content of the samples\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eCalibration curve data for TPC.\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\u003eConcentration ppm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAbsorbance at 765 nm\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.051\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.118\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.173\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.233\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.291\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.348\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.422\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.475\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.550\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.598\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe amount of phenolics present (total) in the \u003cem\u003eM. indica\u003c/em\u003e and \u003cem\u003eT. divaricata\u003c/em\u003e extracts were found to be 62.5 GAE mg/100 mg and GAE 55 mg/100 mg respectively.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec27\" class=\"Section2\"\u003e \u003ch2\u003e3.6 Pharmacological evaluation\u003c/h2\u003e \u003cp\u003eThe anti-oxidant and anti-inflammatory properties, extracts of \u003cem\u003eM. indica\u003c/em\u003e and \u003cem\u003eT. divaricata\u003c/em\u003e were combined in three different ratios. These combined extracts were labelled as C1, C2 and C3 containing 1:1, 1:2 and 2:1 ratio of \u003cem\u003eM. indica\u003c/em\u003e and \u003cem\u003eT. divaricata\u003c/em\u003e respectively.\u003c/p\u003e \u003cdiv id=\"Sec28\" class=\"Section3\"\u003e \u003ch2\u003e3.6.1 DPPH radical scavenging assay\u003c/h2\u003e \u003cp\u003eUsing the DPPH radical scavenging assay method, the combined extracts' antioxidant activity was ascertained. When appropriate reducing chemicals are used to react with DPPH radicals, the radicals lose color stoichiometric to the number of electrons consumed, which may be detected using a 517 nm using spectrophotometer. If DPPH has antioxidant properties, the deep purple color of the molecule becomes less intense. The antioxidant activity was found to be synergistically improved on combining the extracts (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e9\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eInhibition of DPPH radical.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTest solution\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC3\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eM. indica\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eT. divaricata\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e% DPPH inhibition\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e65.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.361\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e82.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.513\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e94.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.702\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e44.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.127\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e35.87\u0026thinsp;\u0026plusmn;\u0026thinsp;1.464\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eIt was found from the result that combining the extracts resulted in synergizing the antioxidant activity when compared to the individual extracts. The highest amount of DPPH inhibition was witnessed in C3 (94.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.702%) where \u003cem\u003eM. indica\u003c/em\u003e and \u003cem\u003eT. divaricata\u003c/em\u003e were mixed in the ratio 2:1 (Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e9\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec29\" class=\"Section3\"\u003e \u003ch2\u003e3.6.2 Anti-inflammatory activity\u003c/h2\u003e \u003cp\u003eThe anti-inflammatory characteristics of the combined extracts were examined using the rat carrageenan-induced paw edema approach. Acute inflammation caused by carrageenan is one of the finest test methods for anti-inflammatory medication evaluation. Having the capacity to extracts and the mixed extracts to inhibit edema on pretreatment was calculated from the paw diameter obtained at different times post treatment (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e10\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eRat paw diameter and edema inhibition.\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=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGroup\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003ePaw thickness (mm)/Edema inhibition (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1h\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2h\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3h\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4h\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.488\u0026thinsp;\u0026plusmn;\u0026thinsp;0.032\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.674\u0026thinsp;\u0026plusmn;\u0026thinsp;0.030\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e0.794\u0026thinsp;\u0026plusmn;\u0026thinsp;0.017\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e0.832\u0026thinsp;\u0026plusmn;\u0026thinsp;0.025\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIbuprofen\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.272\u0026thinsp;\u0026plusmn;\u0026thinsp;0.036 (44.26)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.352\u0026thinsp;\u0026plusmn;\u0026thinsp;0.016 (47.77)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e0.362\u0026thinsp;\u0026plusmn;\u0026thinsp;0.041 (54.41)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e0.216\u0026thinsp;\u0026plusmn;\u0026thinsp;0.023 (74.04)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eM. indica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.420\u0026thinsp;\u0026plusmn;\u0026thinsp;0.056 (13.93)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.446\u0026thinsp;\u0026plusmn;\u0026thinsp;0.036 (33.83)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e0.450\u0026thinsp;\u0026plusmn;\u0026thinsp;0.037 (43.32)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e0.414\u0026thinsp;\u0026plusmn;\u0026thinsp;0.026 (50.24)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eT. divaricata\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.452\u0026thinsp;\u0026plusmn;\u0026thinsp;0.024 (7.38)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.508\u0026thinsp;\u0026plusmn;\u0026thinsp;0.016 (24.63)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e0.524\u0026thinsp;\u0026plusmn;\u0026thinsp;0.009 (34.01)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e0.442\u0026thinsp;\u0026plusmn;\u0026thinsp;0.019 (46.88)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.392\u0026thinsp;\u0026plusmn;\u0026thinsp;0.020 (19.67)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.434\u0026thinsp;\u0026plusmn;\u0026thinsp;0.023 (35.61)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e0.436\u0026thinsp;\u0026plusmn;\u0026thinsp;0.019 (45.09)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e0.402\u0026thinsp;\u0026plusmn;\u0026thinsp;0.048 (51.68)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.372\u0026thinsp;\u0026plusmn;\u0026thinsp;0.027 (23.77)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.422\u0026thinsp;\u0026plusmn;\u0026thinsp;0.024 (37.39)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e0.428\u0026thinsp;\u0026plusmn;\u0026thinsp;0.016 (46.10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e0.376\u0026thinsp;\u0026plusmn;\u0026thinsp;0.030 (54.81)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.352\u0026thinsp;\u0026plusmn;\u0026thinsp;0.029 (27.87)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.406\u0026thinsp;\u0026plusmn;\u0026thinsp;0.032 (39.76)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e0.418\u0026thinsp;\u0026plusmn;\u0026thinsp;0.013 (47.36)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e0.326\u0026thinsp;\u0026plusmn;\u0026thinsp;0.015 (60.82)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"4. Conculsion","content":"\u003cp\u003eThe study concludes that the combination of \u003cem\u003eMangifera indica\u003c/em\u003e and \u003cem\u003eTabernaemontana divaricata\u003c/em\u003e leaf extracts significantly enhances their antioxidant and anti-inflammatory properties compared to the individual extracts. The observed synergistic effects, particularly in the 2:1 ratio of \u003cem\u003eM. indica\u003c/em\u003e to \u003cem\u003eT. divaricata\u003c/em\u003e, suggest that these plant extracts could be effectively utilized in developing natural therapeutics for managing conditions associated with oxidative stress and inflammation. The presence of bioactive compounds such as flavonoids and phenolics likely contributes to these enhanced activities, reinforcing the potential of these plant combinations in therapeutic applications. Research results led to the conclusion that combining the extracts greatly enhanced their anti-inflammatory and antioxidant properties. The greatest increase in activity was specifically observed when two parts \u003cem\u003eM. indica\u003c/em\u003e extract and one part \u003cem\u003eT. divaricata\u003c/em\u003e extract were used together.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u0026nbsp;\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cul type=\"disc\"\u003e\n \u003cli\u003eGanesh Mangesh Walanj, Pranjali Thakare and Nasir Ahmad Sarwary: Conceptualization, methodology, Data collection, analysis, writing \u0026ndash; original draft.\u003c/li\u003e\n \u003cli\u003eAnand Kumar, Dilip KumarTiwari, Abhishek Sharma and Devesh Kumar:\u0026nbsp;Data collection, project administration, writing \u0026ndash; review \u0026amp; editing.\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that there are no competing interests related to this publication.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was conducted in accordance with the ethical standards of the Institutional Review Board of the College of Food Science and Technology, Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Ocean University, Zhanjiang, China, and the Laxmi Narayan College of Pharmacy, Department of Pharmacology, Rajiv Gandhi Proudyogiki Vishwavidyalaya, Bhopal, India. Informed consent was obtained from all participants involved in the study. All procedures performed were in line with the Declaration of Helsinki.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated during and/or analyzed during the current study are available from the corresponding author upon reasonable request, especially for those in need of the data for further research.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eInformed consent was obtained from all individual participants included in the study.\u0026nbsp;Informed consent was obtained from all participants prior to their inclusion in the study. Participants were fully informed about the nature, purpose, and potential risks of the research, and their participation was voluntary.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to publish\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors have given their consent for the publication of this manuscript. Additionally, informed consent was obtained from participants for the publication of any identifying information and data.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAmreen R, and Chaurey M (2021) Evaluation of Estrogenic Potential of Ethanolic and Aqueous Extract of Pitunia Hybrida. J Pharmacol Biomed 5(4):312\u0026ndash;318. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://journals.indexcopernicus.com/api/file/viewByFileId/1388005\u003c/span\u003e\u003cspan address=\"https://journals.indexcopernicus.com/api/file/viewByFileId/1388005\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBanu SK, and Cathrine L (2015) General Techniques Involved in Phytochemical Analysis. Int J Adv Res Chem Sci 2(4):25\u0026ndash;32. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.arcjournals.org/international-journal-of-advanced-research-in-chemical-science/volume-2-issue-4/5\u003c/span\u003e\u003cspan address=\"https://www.arcjournals.org/international-journal-of-advanced-research-in-chemical-science/volume-2-issue-4/5\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEdiriweera MK, Tennekoon KH, and Samarakoon SR (2017) A Review on Ethnopharmacological Applications, Pharmacological Activities, and Bioactive Compounds of Mangifera Indica (Mango). Evidence-Based Complement Altern Med 2017(1):6949835. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1155/2017/6949835\u003c/span\u003e\u003cspan address=\"10.1155/2017/6949835\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhosh P, Poddar S, and Chatterjee S (2021) Morphological Features, Phytochemical and Ethnopharmacological Attributes of Tabernaemontana Divaricata Linn.: A Comprehensive Review. J Pharmacognosy Phytochemistry 10(6):31\u0026ndash;36. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.22271/phyto.2021.v10.i6a.14253\u003c/span\u003e\u003cspan address=\"10.22271/phyto.2021.v10.i6a.14253\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKemisetti D, and Manda S (2018) Synthesis and Comparison of Peg-Ibuprofen and Peg-Ketoprofen Prodrugs by in Vitro and in Vivo Evaluation. J Drug Delivery Ther 8(4):145\u0026ndash;154. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.22270/jddt.v8i4.1743\u003c/span\u003e\u003cspan address=\"10.22270/jddt.v8i4.1743\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKUMAR PRADEEP, SRIVASTAVA V, and KUMAR L (2008) Development of New Reversed-Phase HPLC Method for the Determination of Mangiferin in Mangifera Indica Linn. Biosci Biotechnol Res Asia 5(1):383\u0026ndash;388. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.biotech-asia.org/vol5no1/development-of-new-reversed-phase-hplc-method-for-the-determination-of-mangiferin-in-mangifera-indica-linn/\u003c/span\u003e\u003cspan address=\"https://www.biotech-asia.org/vol5no1/development-of-new-reversed-phase-hplc-method-for-the-determination-of-mangiferin-in-mangifera-indica-linn/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLeyane TS, Jere SW, Houreld NN (2022) Oxidative Stress in Ageing and Chronic Degenerative Pathologies: Molecular Mechanisms Involved in Counteracting Oxidative Stress and Chronic Inflammation. Int J Mol Sci 23(13):7273. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/ijms23137273\u003c/span\u003e\u003cspan address=\"10.3390/ijms23137273\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMirza B et al (2021) Mango (Mangifera Indica L.): A Magnificent Plant with Cancer Preventive and Anticancer Therapeutic Potential. Crit Rev Food Sci Nutr 61(13):2125\u0026ndash;2151. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/10408398.2020.1771678\u003c/span\u003e\u003cspan address=\"10.1080/10408398.2020.1771678\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMishra S, and Jain S (2021) Hepatoprotective and Antioxidant Effect of Delonix Regia Leaf Extract against D Galactosamine Induced Oxidative Stress in Rats. J Pharmacol Biomed 5(3):326. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://journals.indexcopernicus.com/api/file/viewByFileId/1388010\u003c/span\u003e\u003cspan address=\"https://journals.indexcopernicus.com/api/file/viewByFileId/1388010\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePandey M, Debnath M, Gupta S, Chikara SK (2011) Phytomedicine: An Ancient Approach Turning into Future Potential Source of Therapeutics. J Pharmacognosy phytotherapy 3(3):27\u0026ndash;37. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.researchgate.net/publication/215511510_Phytomedicine_An_ancient_approach_turning_into_future_potential_source_of_therapeutics\u003c/span\u003e\u003cspan address=\"https://www.researchgate.net/publication/215511510_Phytomedicine_An_ancient_approach_turning_into_future_potential_source_of_therapeutics\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShah UM et al (2010) Development and Validation of a Simple Isocratic HPLC Method for Simultaneous Estimation of Phytosterols in Cissus Quadrangularis. Indian J Pharm Sci 72(6):753. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.4103/0250-474x.84587\u003c/span\u003e\u003cspan address=\"10.4103/0250-474x.84587\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\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":"laxmi narayan college of pharmacy bhopal","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":"Anti-inflammatory, Anti-oxidant, Tabernaemontana divaricate, Mangifera indica","lastPublishedDoi":"10.21203/rs.3.rs-5344453/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5344453/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eEthnopharmacological Relevance: Tabernaemontana divaricata\u003c/h2\u003e \u003cp\u003e(\u003cem\u003eT. divaricata\u003c/em\u003e) and \u003cem\u003eMangifera indica\u003c/em\u003e (\u003cem\u003eM. indica\u003c/em\u003e) are plants traditionally used in Ayurvedic medicine for their anti-inflammatory and antioxidant properties. \u003cem\u003eT. divaricata\u003c/em\u003e is known for its analgesic and wound-healing effects, while \u003cem\u003eM. indica\u003c/em\u003e is celebrated for its role in managing inflammation and oxidative stress.\u003c/p\u003e\u003ch2\u003eAim of the Study:\u003c/h2\u003e \u003cp\u003eThe study aimed to evaluate the synergistic effects of combined hydroalcoholic extracts from \u003cem\u003eT. divaricata\u003c/em\u003e and \u003cem\u003eM. indica\u003c/em\u003e leaves, focusing on their anti-inflammatory and antioxidant properties.\u003c/p\u003e\u003ch2\u003eMaterials and Methods\u003c/h2\u003e \u003cp\u003eThe leaves were collected, authenticated, and subjected to hydroalcoholic extraction. The extracts were then combined in various ratios and analyzed for phytochemical content. The antioxidant activity was evaluated using the DPPH assay, while anti-inflammatory effects were assessed through the carrageenan-induced rat paw edema model.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eThe combined extracts exhibited significantly enhanced antioxidant and anti-inflammatory activities compared to the individual extracts. The most potent combination, a 2:1 ratio of \u003cem\u003eM. indica\u003c/em\u003e to \u003cem\u003eT. divaricata\u003c/em\u003e, showed the highest DPPH inhibition (94.37%) and maximum edema reduction in the animal model.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eThe combination of \u003cem\u003eT. divaricata\u003c/em\u003e and \u003cem\u003eM. indica\u003c/em\u003e extracts offers a synergistic therapeutic potential, enhancing both antioxidant and anti-inflammatory effects, which supports their traditional use and suggests potential for developing new natural therapeutics.\u003c/p\u003e","manuscriptTitle":"In Vitro and In-Vivo Analysis of the Combined Extract of Tabernaemontana divaricata and Mangifera indica Leaves for Enhanced Anti-Inflammatory and Antioxidant Properties","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-10-29 08:14:40","doi":"10.21203/rs.3.rs-5344453/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":"da60f254-9085-49ba-b228-e10db3173b06","owner":[],"postedDate":"October 29th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":39483484,"name":"Clinical Pharmacology"}],"tags":[],"updatedAt":"2024-10-29T08:14:40+00:00","versionOfRecord":[],"versionCreatedAt":"2024-10-29 08:14:40","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5344453","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5344453","identity":"rs-5344453","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}