Phytochemical Analysis and Evaluation of the Anti-Inflammatory and Antioxidant Activity of Diverse Extracts of Millettia peguensis Leaves

Phytochemical Analysis and Evaluation of the Anti-Inflammatory and Antioxidant Activity of Diverse Extracts of Millettia peguensis Leaves | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Phytochemical Analysis and Evaluation of the Anti-Inflammatory and Antioxidant Activity of Diverse Extracts of Millettia peguensis Leaves Anitha Nandagopal, Nemala Appala Raju This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4748245/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 Millettia peguensis is a plant native to Southeast Asia belonging to the Fabaceae family. Leaf extracts were prepared using chloroform, methanol, and water. The carrageenan-induced paw edema model in rats was employed to assess anti-inflammatory activity. Paw thickness was measured over time to quantify edema. All extracts exhibited a reduction in paw edema compared to the control group. The water extract demonstrated a significant reduction at the third hour post-injection. Both methanol and chloroform extracts displayed significant inhibition by the fourth hour, but their efficacy was lower than the positive control group treated with indomethacin. By the fifth hour, both water and methanol extracts produced results comparable to indomethacin. The study assessed DPPH antioxidant activity of extracts from using chloroform, methanol, and distilled water. Methanol and distilled water extracts showed higher antioxidant activity compared to chloroform extract, suggesting the significance of polar compounds. The findings highlight the impact of solvent choice on extracting bioactive antioxidants. Phytochemical analysis suggested the presence of bioactive compounds such as flavonoids, tannins, saponins, and phenolic compounds. Methanol and water extracts showed higher antioxidant activity due to the presence of phenolic compounds, which may be responsible for the observed anti-inflammatory activity. Millettia peguensis anti-inflammatory activity carrageenan-induced paw edema indomethacin DPPH Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Millettia peguensis , or Moulmein Rosewood, is a small, deciduous tree from the Fabaceae family, valued for its ornamental beauty and mauve, pea-like flowers [ 1 ]. Species in the Millettia genus, like M. conrauai, are known for their insecticidal, molluscicidal, and piscicidal properties [ 2 ]. In Africa, especially Cameroon, Millettia species are used to treat intestinal parasites in children, colic, and boils. In some African regions, particularly Cameroon, various Millettia species are employed by communities to treat intestinal parasites in children and as remedies for colic and boils. Several phytoconstituents were isolated from Millettia peguensis , notably flavonoids such as pongamol, lanceolatin-B, kanjone, milletenone, ovaliflavanone-A, ovalitenone, and pongaglabol [ 3 ]. Pongamol demonstrated larvicidal activity against Aedes aegypti [ 4 ], while pongaglabol showed antioxidant properties [ 5 ]. Additionally, other phytochemicals from M. peguensis include 9-methylheptadecane, eicosane, heneicosane, dodecyl hexyl ester, and 2-bromododecane. Lanceolatin-B exhibited antibacterial and anti-inflammatory activities, and pongaglabol has been reported for antiulcer activity [ 6 ]. The genus Millettia is known for its insecticidal, pesticidal, antimalarial, and antioxidant potentials [ 7 ]. Figure 1 depicts the leaf and flower of Millettia peguensis. Experimental Materials Collection and Identification of Plant Materials Millettia peguensis leaves acquired from Balmukund Devkaran Sadan, Hyderabad, was meticulously examined and subsequently approved by a qualified botanist. This rigorous verification ensures the authenticity and scientific accuracy of the specimen, contributing significantly to the credibility of ongoing research and studies. The collected leaves were dried in the shade, ground into coarse powder, and stored in an airtight container until needed for experiments. Ethics Approval and Consent to Participate The authors confirm that all methods were carried out in accordance with relevant guidelines and regulations of NMPB. The collection of the plants used in the study complies with local or national guidelines with no need for further affirmation. Millettia peguensis was collected in Balmukund Devkaran Sadan, Hyderabad. The plant material was identified by Dr. V. B. Reddy and a voucher specimen was deposited at Dept. of Botany, OU Hyderabad with voucher ID OUAS-227. Experimental Animals Healthy Swiss albino rats of both sexes, weighing 150-210 g, were purchased from an animal supplier. The rats were acclimatized for one week in standard cages under controlled conditions: a constant temperature of 22 ± 1 °C, relative humidity of 65 ± 5%, and a 12-hour light-dark cycle. They had free access to a standard rat diet and water. Ethical Approval and Participant Consent The experimental protocol was approved by the institutional animal ethical committee of MRM College of Pharmacy Hyderabad, under approval number IAEC/MRM/2023/05. This ensures the study complies with ethical standards, guaranteeing proper care and use of animals in research, and upholding the integrity and validity of the experimental procedures. Chemicals and Reagents The organic solvents used for plant material extraction included methanol and chloroform (Rankem Chemicals, India). Other chemicals and reagents used were: 1% alcoholic α-naphthol solution, indomethacin, and carrageenan (Sigma-Aldrich, India) Sodium carbonate (Rankem, India) Acetic acid, acetic anhydride, concentrated sulfuric acid (Rankem, India) Ammonia, Benedict’s reagent, Biuret reagent, diluted hydrochloric acid, ferric chloride, Folin-Ciocalteu phenol reagent, Hager’s reagent, Mayer’s reagent, pyridine, sodium hydroxide solution, sodium nitroprusside solution, and Sudan Red III reagent (Sigma Aldrich Chemicals, India) DPPH (2,2-diphenyl-1-picrylhydrazyl) powder Ethanol (analytical grade) Methanolic extract of the sample specimen Ascorbic acid (standard antioxidant) Apparatus and Equipment Laboratory Glassware Various laboratory glassware items were utilized, including beakers, conical flasks, round bottom flasks, a grinder for plant material, measuring cylinders, porcelain dishes, heating mantles, filter papers, test tubes, stirring glass rods, micro hematocrit tubes, a digital weighing balance, and a water bath. Specific Instruments Specific instruments included a Soxhlet extractor with a condenser, a hot air oven, a desiccator, digital calipers, a microscope, a rotavapour, insulin syringe needles and a metal gavage needle. Preparation of Plant Extracts The plant leaves were dried in the shade and ground into coarse powder. Approximately 230 g of this powder was placed in a Soxhlet extraction apparatus [8]. Extraction was performed using chloroform, methanol, and water, each at twice the weight of the dried plant powder. Solvents wetted the powder and filled the round bottom flask, which was heated to evaporate the solvent. The vapor passed through the plant material, condensed, and dripped back onto the powder. Extraction continued until the solvent in the siphon lost its color. The extract was then filtered to remove impurities. Following Soxhlet extraction with chloroform and methanol, the remaining sample underwent distilled water extraction using maceration. The sample was soaked in distilled water for three days with periodic agitation and then strained. Chloroform extracted non-polar compounds, methanol extracted polar compounds, and water extracted highly polar compounds. Concentration of Extracts Each extract underwent concentration using a rotary evaporator with specific conditions: the chloroform extract was concentrated at 62°C under 475 mbar pressure, the methanol extract at 65°C under 335 mbar pressure, and the distilled water extract at 100°C under 72 mbar pressure. After concentration, the extracts were stored in a desiccator containing calcium chloride to eliminate moisture. Finally, the concentrated extracts were transferred into airtight containers and stored at 4°C. Preliminary Phytochemical Analysis All extracted samples were subjected to preliminary phytochemical analysis to qualitatively identify various compounds present. The screening aimed to detect alkaloids, flavonoids, terpenoids, phenolic compounds, tannins, glycosides, saponins, fixed oils and fats, carbohydrates, and proteins within the plant extracts [9]. Experimental Design for Rat Paw Edema Study [10] Thirty healthy Swiss albino rats weighing between 150-210 g were chosen for the experiment. The rats were randomly divided into five groups, each consisting of six rats. These groups included a positive control, a negative control, and three experimental groups treated with previously extracted plant extracts. The chloroform, methanol, and distilled water extracts were administered orally at a dose of 100 mg/kg to their respective groups. Additionally, standard drug indomethacin and normal saline were administered orally as controls. Figure 2 depicts the oral administration of extracts in rats. Preparations of the extracts were tailored to each rat's weight prior to administration. The chloroform extract was suspended in an emulsion of olive oil, distilled water, and acacia gum (in a ratio of 4:2:1) due to its insolubility in water. The methanol and distilled water extracts were dissolved in distilled water before administration. Table 1 provides an overview of the experimental groups and their respective treatments. Table-1: Description of experimental design groups Groups (n=6) Experimental design groups Treatment 1 Positive control Carrageenan + 10 mg/kg Indomethacin 2 Chloroform Carrageenan + 100 mg/kg Chloroform extract 3 Methanol Carrageenan + 100 mg/kg Methanol extract 4 Distilled water Carrageenan + 100 mg/kg Distilled water extract 5 Negative control Carrageenan + Normal Saline Each group consisted of 6 rats (n=6), and these treatments were administered to evaluate the anti-inflammatory effects using a carrageenan-induced paw edema model. Table1 provides an overview of the experimental groups in the study, detailing their respective treatments: the positive control received carrageenan injection followed by 10 mg/kg of indomethacin; the chloroform, methanol, and distilled water extract groups received carrageenan injection followed by 100 mg/kg of their respective extracts; and the negative control received carrageenan injection followed by normal saline. Six rats were randomly placed into labeled cages, with each cage representing a different experimental group. Paw thickness was measured for each rat every 60 minutes from 0 to 300 minutes during the experiment. After the experiment, the percentage of edema inhibition was calculated for each set of data. The anti-inflammatory potential of various extracts of Millettia peguensis was assessed using the carrageenan-induced hind paw edema model in albino rats. Indomethacin was administered as a standard treatment in the positive control group at a dosage of 10 mg/kg. Each extract was administered at a dose of 100 mg/kg body weight to separate groups of rats, one hour before inducing inflammation by injecting carrageenan into the left hind paw plantar tissue. Carrageenan was prepared by dissolving 1% solution in normal saline to create a 0.1 mL injection. Paw thickness was measured before carrageenan injection and hourly thereafter for five hours using a digital caliper, measuring at the dorsa-plantar axis at the metatarsal level. The anti-inflammatory activity was quantified as the percentage of edema inhibition in the positive control and extract-treated groups compared to the negative control group. The percentage of edema inhibition was calculated using the formula: Where Tt is the mean paw thickness of the treated group rats and Tc is the mean paw thickness of the control group rats at the same time interval. Administration of drugs and experimental compounds in rats Proper handling and restraint techniques were employed to minimize stress during procedures. The V-grip method, following [11] gently gripped the rat's head with knuckles on the jaw bone and secured the thorax under both forelimbs. This method, recommended for the dominant hand, supports the rat's lower body or rests it on the handler's chest for comfort, as suggested by Moore [12]. Oral gavage involved administering substances directly into the stomach using a specialized blunt-ended gavage needle with a ball tip, ensuring precise dosing and rapid absorption compared to dietary methods. Maximum volumes were based on individual rat weights, and needle lengths were adjusted to avoid injury or aspiration. The syringe was prepared with the substance, and the needle surface wiped clean to prevent residue affecting taste. Rats were gently restrained using the V-grip method, the needle inserted into the back of the throat along the mouth's left side and roof, and the rat's head tilted back to straighten the esophagus. The needle was advanced until the pre-marked line reached the mouth, with occasional slight twists to pass the epiglottis. The substance was slowly injected into the stomach, and the needle withdrawn carefully before the rat was returned to its cage. Intraplantar Injection The carrageenan solution was injected intraplantarly into rats, a technique blending aspects of subcutaneous and intradermal injections widely used in inflammation, pain, and immunization studies [13]. Inflammation was induced exclusively in the left hind paw with an injection volume not exceeding 0.10 mL per rat. Before injection, the site was sterilized with a 70% isopropyl alcohol swab. A needle size of 25G or smaller delivered 0.1 mL of 1% carrageenan solution in normal saline just beneath the epidermis, at a shallow angle. Post-injection, the needle was gently withdrawn with light pressure on the site to prevent leakage. Firm pressure was briefly applied to confirm successful injection, evidenced by the formation of a distinct bleb at the site. Figure 3 shows the measurement of paw thickness with a digital Vernier caliper. Statistical Analysis Data were presented as mean ± standard deviation (SD) and analyzed using Statistical Package for the Social Sciences (SPSS) version 23.0. One-way analysis of variance (ANOVA) followed by Tukey's honestly significant difference (HSD) post hoc test was employed for statistical comparisons, with significance set at p < 0.05. Antioxidant Activity of Methanolic Extract of Millettia peguensis [14] Preparation of DPPH Solution: Dissolved 4 mg of DPPH in 100 mL of ethanol to prepare a 0.1 mM DPPH solution. Protected the solution from light by covering the container with aluminum foil or using an amber bottle. Preparation of Sample Extract Solutions: Prepared different concentrations of the methanolic extract of the sample specimen (e.g., 10, 25, 50, 75, 100 µg/mL) by diluting the stock extract with methanol. Preparation of Standard Solution: Prepared a standard solution of ascorbic acid (e.g., 10, 25, 50, 75, 100 µg/mL) in ethanol or methanol. Reaction Mixture Preparation: In test tubes or a 96-well microplate, 1 mL of DPPH solution was added to 1 mL of each concentration of the sample extract. For the standard, 1 mL of DPPH solution was added to 1 mL of each concentration of the ascorbic acid solution. Prepared a control by adding 1 mL of DPPH solution to 1 mL of ethanol (without the sample extract or standard). Incubation: Vortexed the mixtures to ensure thorough mixing. Incubated the mixtures at room temperature in the dark for 30 minutes. Measurement of Absorbance: Measured the absorbance of each reaction mixture at 517 nm using a UV-Vis spectrophotometer. The decrease in absorbance indicates the scavenging of DPPH free radicals by the antioxidants present in the sample extract or standard. Calculation of DPPH Radical Scavenging Activity: Plotting the Results: Plotted the scavenging activity (%) against the concentrations of the sample extract and standard. Determined the IC50 value, which was the concentration of the sample required to scavenge 50% of the DPPH radicals. RESULTS AND DISCUSSION Extraction of Millettia peguensis Leaves The extraction of Millettia peguensis leaves involved the sequential use of three solvents: chloroform, methanol, and distilled water, chosen based on their varying polarities to extract secondary metabolites of interest. Initially, 230 g of dried leaf powder was subjected to extraction using a Soxhlet extractor with chloroform and methanol, followed by maceration with distilled water. The choice of solvent followed a gradient from low to high polarity. The percentage yield of each solvent extraction was calculated using the formula: The extracted masses, percentage yields, colors, and consistencies of the Millettia peguensis leaf extracts are summarized in Table 2. Table 2: Extraction metrics of Millettia peguensis Leaves Extracts Extract Weight (g) Yield (%) Color Consistency Chloroform 19.12 8.31 Grey-black Solid Methanol 43.75 19.02 Grey-black Semisolid Water 12.62 5.49 Dark brown Semisolid This table shows plant extracts using chloroform, methanol, and water, with yields of 8.31%, 19.02%, and 5.49%, respectively. Chloroform and methanol extracts were grey-black, with chloroform being solid and methanol semisolid. The water extract was dark brown and semisolid, reflecting differences in solvent polarity and extracted compounds. Phytochemical analysis of the extracts: The Millettia peguensis leaf extracts were tested for phytochemicals. Results were shown below. Table 3: Results of phytochemical analysis of Millettia peguensis leaf extracts Constituent Types of test Chloroform Methanol Distilled water Flavonoids Alkaline reagent test + ++ +++ Terpenoids Salkowski reaction + + - Liebermann-Burchard reaction + + - Tannins 5% Ferric chloride - + ++ Acetic acid - - - Alkaloids Mayer’s test - + - Hager’s test - + - Glycosides Legal’s test - - - Modified Borntrager’s test ++ + ++ Phenolic compounds Folin-Ciocalteau Test - ++ +++ 5% Ferric chloride - + ++ Saponins Foam test - + + Carbohydrates Molisch’s test ++ + + Benedict’s test + + + Protein Biuret test - - - Xanthoprotein test - - - Fats and oils Sudan red III reagent test + - - - : Not present + : Present in low concentration ++ : Present in high concentration Table 3 compares phytochemical constituents in an extract using chloroform, methanol, and distilled water. Flavonoids are more pronounced in methanol and water, indicated by strong alkaline reagent reactions. Terpenoids are detected in chloroform and methanol but absent in water. Tannins show higher presence in methanol and water via ferric chloride tests. Alkaloids are solely detected in methanol. Glycosides are minimal, observed only in chloroform with moderate response in Borntrager’s test. Phenolic compounds are prominent in methanol and water, notably with Folin-Ciocalteau and ferric chloride tests. Saponins are positive in methanol and water. Carbohydrates are detected in all solvents. Protein, fats, and oils are minimally detected, predominantly in chloroform. Preliminary paw thickness of normal albino rats [15] Initial paw thickness of normal albino rats was measured before inducing edema in 30 randomly grouped rats, divided into five groups with six rats each. Paw thickness was assessed using a calibrated digital Vernier caliper along the dorsa-plantar axis at the metatarsal level, as depicted in Table 4 shows the initial mean paw thickness for each group. Analysis indicated no significant difference in initial paw thickness between the control and experimental groups, confirming uniformity among all groups before inducing edema. Table 4 presents initial paw thickness measurements (mm) for experimental groups of albino rats: positive control (indomethacin), chloroform extract, methanol extract, distilled water extract, and negative control (normal saline). Each value is mean ± standard deviation (S.D.) from six rats/group. Table 4: Initial Paw Thickness in Various Experimental Groups Albino Wistar Group Initial paw thickness (mm) Positive control, Indomethacin 2.27 ± 0.19 Chloroform 2.50 ± 0.48 Methanol 2.48 ± 0.32 Distilled water 2.26 ± 0.12 Negative control, normal saline 2.46 ± 0.19 Statistical analysis (p > 0.05) indicated no significant differences in initial paw thickness, ensuring uniformity at the study's outset. These measurements serve as a baseline for comparing subsequent paw thickness post-edema induction, facilitating evaluation of treatment effects on inflammation. Evaluation of anti-inflammatory activity of various extracts from Millettia peguensis leaves on rat paw edema induced by carrageenan The study assessed anti-inflammatory effects of Millettia peguensis leaf extracts on carrageenan-induced paw edema in rats. Extracts in chloroform, methanol, and water, alongside indomethacin (standard drug), were orally administered at 100 mg/kg and 10 mg/kg doses, one-hour pre-injection. Paw thickness was measured hourly post-injection over five hours to evaluate inflammation impact. Results, including paw thickness and percentage inhibition of inflammation, are summarized in Table 5. ANOVA with Tukey’s HSD test indicated significant differences (p < 0.05) in paw thickness among treatment groups. Table 5 Data shows that extracts of Millettia peguensis and indomethacin reduce rat paw thickness and edema at different times. Group Time interval 1 st hour 2 nd hour 3 rd hour 4 th hour 5 th hour 1 3.42 ± 0.46 3.68 ± 0.47 e 3.46 ± 0.51 b, e 3.12 ± 0.45 b, c, e 2.94 ± 0.50 b, e (14.26) (15.20) (22.99) (30.33) (33.61) 2 4.12 ± 0.41 4.37 ± 0.28 4.35 ± 0.30 a 4.07 ± 0.27 a, e 3.78 ± 0.30 a, e (1.39) (3.17) (8.74) (16.08) (21.56) 3 4.04 ± 0.41 4.19 ± 0.44 4.24 ± 0.45 3.99 ± 0.40 a, e 3.58 ± 0.35 e (2.95) (6.32) (10.61) (17.41) (24.55) 4 4.01 ± 0.64 4.09 ± 0.73 3.74 ± 0.81 e 3.60 ± 0.83 e 3.31 ± 0.74 e (3.17) (8.09) (18.62) (23.32) (28.37) 5 4.19 ± 0.26 4.55 ± 0.27 a 4.87 ± 0.16 a, d 5.08 ± 0.21 a, b, c, d 5.16 ± 0.28 a, b, c, d (0.00) (0.00) (0.00) (0.00) (0.00) Note: Data represent the mean ± SD of 6 animals in each group. Values within parenthesis indicate percentage of inhibition (%). a: Significant compared to group 1 (indomethacin; p<0.05). b: Significant compared to group 2 (chloroform extract; p<0.05). c: Significant compared to group 3 (methanol extract; p<0.05). d: Significant compared to group 4 (distilled water extract; p<0.05). e: Significant compared to group 5 (negative control; p<0.05). In rats, intraplantar carrageenan injection caused paw volume to increase over time, peaking at 5.66 ± 0.31 mm by the fifth hour post-injection. Initially, chloroform and methanol extracts did not significantly affect paw swelling until after the fourth hour. Indomethacin and distilled water extract demonstrated significant anti-inflammatory effects starting from the second and third hour post-injection, respectively. Indomethacin treatment exhibited the highest paw edema inhibition (33.61%) compared to other treatments, significantly reducing swelling starting from the second hour (peak: 3.68 ± 0.47 mm) compared to the negative control group. By the third hour post-injection, chloroform extract, distilled water extract, and indomethacin treatment groups showed decreasing paw swelling. Distilled water extract significantly reduced swelling compared to the control group (p<0.05), while chloroform extract (4.35 ± 0.30 mm) did not show significant effects at the same time point. The paw thickness in the methanol group decreased from 4.24 ± 0.45mm at the third hour post-carrageenan injection to 3.99 ± 0.40 mm at the fourth hour. Paw thickness continued to decrease in other groups, with indomethacin treatment showing significantly greater reduction compared to chloroform and methanol extracts. Distilled water extract exhibited the highest inhibition percentage (28.37%), followed by methanol extract (24.55%), both significantly different from the negative control group. Chloroform extract also showed significant edema suppression (21.56%) compared to the negative control, albeit less effective than indomethacin. Phytochemical analysis revealed that methanol extract contained more active constituents than distilled water extract, specifically alkaloids and terpenoids. However, distilled water extract showed superior edema inhibition, possibly due to higher concentrations of constituents detected, including flavonoids, tannins, and phenolic compounds. Chloroform extract exhibited the lowest variety of active constituents and demonstrated relatively weaker edema inhibition compared to distilled water and methanol extracts. Therefore, at 100 mg/kg, distilled water extract of Millettia peguensis reduces carrageenan-induced edema similarly to the potent anti-inflammatory drug indomethacin in a time-dependent manner. The swelling level was measured by recording the increase in paw thickness at different times, shown in Table 6, with data presented as the swelling level ± standard deviation. Comparative Analysis of Swelling Degrees Over Five Hours Table 6 presents a comparative analysis of the degree of swelling (mm) in various groups over a five-hour period. Each group, including positive and negative controls, and chloroform, methanol, and distilled water-treated samples, were evaluated for swelling at hourly intervals. The values are expressed as means ± standard deviation, allowing for precise assessment of each treatment's anti-inflammatory potential. Table 6: Level of swelling at different time intervals Group Degree of swelling (mm) 1 st hour 2 nd hour 3 rd hour 4 th hour 5 th hour Positive control 1.33 ±0.34 1.58 ±0.35 1.36 ±0.47 1.02 ±0.40 0.84 ±0.44 Chloroform 1.78 ±0.30 2.04 ±0.28 2.02 ±0.38 1.74 ±0.27 1.45 ±0.20 Methanol 1.73 ±0.53 1.88 ±0.57 1.93 ±0.50 1.68 ±0.48 1.27 ±0.44 Distilled water 1.92 ±0.68 2.00 ±0.77 1.65 ±0.86 1.51 ±0.88 1.22 ±0.79 Negative control 1.90 ±0.36 2.26 ±0.37 2.58 ±0.30 2.79 ±0.32 2.87 ±0.38 Treatment-specific Observations and Temporal Trends The positive control group shows a gradual reduction in swelling from 1.33 ± 0.34 mm at the first hour to 0.84 ± 0.44 mm at the fifth hour, indicating effective anti-inflammatory activity. Conversely, the negative control group exhibits a continuous increase in swelling, from 1.90 ± 0.36 mm at the first hour to 2.87 ± 0.38 mm by the fifth hour, highlighting the baseline inflammatory response without treatment. In the chloroform-treated group, swelling initially increases from 1.78 ± 0.30 mm, peaking at 2.04 ± 0.28 mm in the second hour, and then declines to 1.45 ± 0.20 mm by the fifth hour, suggesting a moderate anti-inflammatory effect. The methanol-treated group shows similar trends, with initial swelling of 1.73 ± 0.53 mm, peaking at 1.93 ± 0.50 mm at the third hour, and reducing to 1.27 ± 0.44 mm at the fifth hour. Distilled Water Treatment and Comparative Efficacy The distilled water-treated group shows initial swelling of 1.92 ± 0.68 mm, peaking at 2.00 ± 0.77 mm at the second hour, and declining to 1.22 ± 0.79 mm by the fifth hour. This pattern indicates that distilled water has moderate anti-inflammatory properties, comparable to methanol and slightly better than chloroform. Antioxidant activity of methanolic extract of Millettia peguensis by DPPH method The table-7 presents the in vitro antioxidant activity of different concentrations of an extract using the DPPH method. The % inhibition of DPPH free radicals was measured at concentrations ranging from 5 to 60 μl/ml. As the concentration increases, the % inhibition also increases, indicating higher antioxidant activity. At the lowest concentration (5 μl/ml), the % inhibition values are 16.18%, 20.23%, and 29.84% for the different extracts tested. As the concentration increases to 60 μl/ml, the % inhibition reaches 81.07%, 89.22%, and 99.74%, respectively. This trend demonstrates a dose-dependent increase in antioxidant activity for all extracts tested, with the highest concentration showing the greatest inhibition of DPPH radicals which were shown in Figure 4. Table 7: Data of Antioxidant activity of Various extracts of Millettia peguensis Concentration [μl/ml]) % Inhibition by Chloroform extract % Inhibition by Methanolic extract % Inhibition by Distilled water extract 5 16.18 20.23 29.84 10 22.89 33.46 42.10 15 32.53 36.84 50.16 20 42.51 43.58 59.35 25 42.00 45.14 64.35 30 52.15 57.47 74.84 35 56.11 67.96 76.29 40 60.93 73.67 83.39 45 66.95 74.97 87.90 50 72.98 83.01 91.94 55 72.46 87.42 97.42 60 81.07 89.22 99.74 Conclusion In conclusion, Millettia peguensis extracts effectively reduced inflammation in carrageenan-induced rat paw swelling. The extracts, especially distilled water, show significant anti-swelling effects comparable to indomethacin. Phytochemical analysis revealed flavonoids, saponins, phenolic compounds, tannins, and terpenoids, suggesting potential benefits for managing inflammation-related conditions like fever and pain. All extracts demonstrated increasing % inhibition of DPPH radicals with higher concentrations. Methanolic and distilled water extracts consistently show higher inhibition rates compared to chloroform extract, particularly notable at higher concentrations (50-60 μl/ml), suggesting their stronger antioxidant potential. Future research should explore the mechanisms underlying Millettia peguensis's anti-inflammatory actions. Continued studies and clinical trials are needed to validate their effectiveness in inflammation reduction, highlighting natural phytoconstituents as promising molecules for drug development. Declarations Author Contribution Both authors, Dr. Nemala Appala Raju and Dr. Anitha Nandagopal, contributed equally to this work. Dr. Nemala Appala Raju and Dr. Anitha Nandagopal were involved in the conceptualization, methodology, formal analysis, and investigation of the research. They jointly handled resources and data curation. The original draft was written by both authors, and both contributed to the review and editing process. Visualization of the results was performed by Dr. Nemala Appala Raju and Dr. Anitha Nandagopal. Supervision and project administration were shared responsibilities between the authors. Both authors have read and agreed to the published version of the manuscript. Data Availability The datasets used and/or analysed during the current study available from the corresponding author on reasonable request References Manikandan G, Vimala Rani A. GC-MS analysis of phytochemical constituents in the petroleum ether leaf extracts of Millettia peguensis . Int Res J Pharm. 2017; 8 (9):144–50. Packiyalakshmi D, Athilakshmi P, Gayathri S, Karthiga P, Thiri Bhuvaneswari R, Manikandan G. Antimicrobial potential of different solvents leaf extract of Millettia peguensis against selected pathogens. Pharma Inno J. 2017; 8(1): 109–24. Ganapaty S, Pushpalata V, Gella JB, Naidu KC, Peter WG. Waterman flavonoids from Millettia peguensis Ali (Fabaceae) Biochem Syst Ecol. 1998; 26(1): 125–26. 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In: Pharmacological assays of plant-based natural products. Prog Drug Res. 2016;71. Springer, Cham. Winter CA, Risley EA, Nuss GW. Carrageenin-induced edema in hind paw of the rat as an assay for antiinflammatory drugs. Proceed Soc Exp Bio Med. 1962;111(3):544–47. Kemp RW. Chapter 3: Handling and restraint, In The Laboratory Rats (Handbook of Experimental Animals), 2000;31–41, Cheshire, United Kingdom, Academic Press. Moore, S. 2015, Standard Operating Procedure - Rodent Handling. https://carleton.ca/animalethics/wp-content/uploads/EXP-07-Rodent-Handling.pdf (accessed 2 December 2017). Kamala T. Hock immunization: A humane alternative to mouse footpad injections. J Immu Meth. 2007, 328(1–2): 204–14. Dontha S. A Review on antioxidant methods. Asian J Pharm Cli Res. 2016; 9(8):14–32. Modi F, Varia R, Patel J. Anti-inflammatory activity of ellagic acid in carrageenan-induced paw edema in rats. Haryana Vet. 2023; 62(SI-2): 18–21. Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4748245","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":340806513,"identity":"95238b25-70d4-42bb-a7f2-f07f129a6707","order_by":0,"name":"Anitha Nandagopal","email":"","orcid":"","institution":"Sultan-ul-Uloom College of Pharmacy","correspondingAuthor":false,"prefix":"","firstName":"Anitha","middleName":"","lastName":"Nandagopal","suffix":""},{"id":340806514,"identity":"37294af9-0fdc-49c4-8b35-16eab90a65d8","order_by":1,"name":"Nemala Appala Raju","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA50lEQVRIiWNgGAWjYHCCBBAhw8DeABNgbMChElULDwPPAeK1MEC0SCQQ6Srz9gPPHnzMsePhn/n8mcTPHQzy/A3MbQ/waZE5k5BuOHNbMo/E7Rwzyd4zDIYzDjC2G+DTIsGQkCbNu42Zh+F2Dps0YxsD4wYGxjYJvFr4H6RJ/91WzyN/8/gzkBZ7wlokgLYwbjvMY3CDwQykJZEILQ/SJHu3HecxPJNjbNl7RiJ5xmGCDstJk/i5rVpO7vjxhzd+7rCx7W9vf4ZXCzBGEhBsxgagYmb86oGA/QCyFoLKR8EoGAWjYAQCAK3WQs/fJDK+AAAAAElFTkSuQmCC","orcid":"","institution":"Sultan-ul-Uloom College of Pharmacy","correspondingAuthor":true,"prefix":"","firstName":"Nemala","middleName":"Appala","lastName":"Raju","suffix":""}],"badges":[],"createdAt":"2024-07-16 08:36:22","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4748245/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4748245/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":62952733,"identity":"491159d5-c25c-4ef8-a98c-8117fe6df219","added_by":"auto","created_at":"2024-08-21 11:36:49","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":39615,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eMillettia peguensis\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e leaf and Flower\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4748245/v1/705ab1d85cdef1df28749c72.jpg"},{"id":62950613,"identity":"9080895d-7ffb-4829-a14f-67a9a65853c3","added_by":"auto","created_at":"2024-08-21 11:12:49","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":26562,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eOral administration of Extracts in Rats\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4748245/v1/630c0257f9533f23ade61bc8.jpg"},{"id":62951136,"identity":"9ca52452-f0db-434a-94df-ba73b9410b7b","added_by":"auto","created_at":"2024-08-21 11:20:49","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":22645,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePaw thickness measurement with a Digital Vernier caliper\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4748245/v1/e44b35c829f178819b2f8295.jpg"},{"id":62951994,"identity":"54c8d0ab-e070-4985-88f8-639d6b5ed0d6","added_by":"auto","created_at":"2024-08-21 11:28:49","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":56409,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAntioxidant activity of Various extracts of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eMillettia peguensis\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4748245/v1/41fc4d8dce2f344c7d6f5a7f.jpg"},{"id":71944805,"identity":"79bda89d-da66-4b61-b681-a4464421c4de","added_by":"auto","created_at":"2024-12-20 03:01:43","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1147739,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4748245/v1/d326b835-ed87-4ecd-b47a-696d31989a97.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Phytochemical Analysis and Evaluation of the Anti-Inflammatory and Antioxidant Activity of Diverse Extracts of Millettia peguensis Leaves","fulltext":[{"header":"Introduction","content":"\u003cp\u003e \u003cem\u003eMillettia peguensis\u003c/em\u003e, or Moulmein Rosewood, is a small, deciduous tree from the Fabaceae family, valued for its ornamental beauty and mauve, pea-like flowers [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Species in the Millettia genus, like M. conrauai, are known for their insecticidal, molluscicidal, and piscicidal properties [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. In Africa, especially Cameroon, Millettia species are used to treat intestinal parasites in children, colic, and boils. In some African regions, particularly Cameroon, various Millettia species are employed by communities to treat intestinal parasites in children and as remedies for colic and boils. Several phytoconstituents were isolated from \u003cem\u003eMillettia peguensis\u003c/em\u003e, notably flavonoids such as pongamol, lanceolatin-B, kanjone, milletenone, ovaliflavanone-A, ovalitenone, and pongaglabol [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Pongamol demonstrated larvicidal activity against Aedes aegypti [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e], while pongaglabol showed antioxidant properties [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Additionally, other phytochemicals from \u003cem\u003eM. peguensis\u003c/em\u003e include 9-methylheptadecane, eicosane, heneicosane, dodecyl hexyl ester, and 2-bromododecane. Lanceolatin-B exhibited antibacterial and anti-inflammatory activities, and pongaglabol has been reported for antiulcer activity [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. The genus \u003cem\u003eMillettia\u003c/em\u003e is known for its insecticidal, pesticidal, antimalarial, and antioxidant potentials [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Figure\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e depicts the leaf and flower of \u003cem\u003eMillettia peguensis.\u003c/em\u003e\u003c/p\u003e \u003cp\u003e \u003c/p\u003e "},{"header":"Experimental","content":"\u003cp\u003eMaterials\u003c/p\u003e\u003cp\u003e \u003cem\u003eCollection and Identification of Plant Materials\u003c/em\u003e \u003c/p\u003e\u003cp\u003e\u003cem\u003eMillettia peguensis\u003c/em\u003e leaves acquired from Balmukund Devkaran Sadan, Hyderabad, was meticulously examined and subsequently approved by a qualified botanist. This rigorous verification ensures the authenticity and scientific accuracy of the specimen, contributing significantly to the credibility of ongoing research and studies. The collected leaves were dried in the shade, ground into coarse powder, and stored in an airtight container until needed for experiments.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eEthics Approval and Consent to Participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors confirm that all methods were carried out in accordance with relevant guidelines and regulations of NMPB. The collection of the plants used in the study complies with local or national guidelines with no need for further affirmation. \u003cstrong\u003e\u003cem\u003eMillettia peguensis\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003ewas collected in\u0026nbsp;Balmukund Devkaran Sadan, Hyderabad. The plant material was identified by Dr. V. B. Reddy and a voucher\u0026nbsp;specimen was deposited at Dept. of Botany, OU Hyderabad with voucher ID OUAS-227.\u003c/p\u003e\n\u003ch4\u003eExperimental Animals\u003c/h4\u003e\n\u003cp\u003eHealthy Swiss albino rats of both sexes, weighing 150-210 g, were purchased from an animal supplier. The rats were acclimatized for one week in standard cages under controlled conditions: a constant temperature of 22 ± 1 °C, relative humidity of 65 ± 5%, and a 12-hour light-dark cycle. They had free access to a standard rat diet and water.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eEthical Approval and Participant Consent\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe experimental protocol was approved by the institutional animal ethical committee of MRM College of Pharmacy Hyderabad, under approval number IAEC/MRM/2023/05. This ensures the study complies with ethical standards, guaranteeing proper care and use of animals in research, and upholding the integrity and validity of the experimental procedures.\u003c/p\u003e\n\u003ch4\u003eChemicals and Reagents\u003c/h4\u003e\n\u003cp\u003eThe organic solvents used for plant material extraction included methanol and chloroform (Rankem Chemicals, India). Other chemicals and reagents used were:\u003c/p\u003e\n\u003cul type=\"disc\"\u003e\n \u003cli\u003e1% alcoholic α-naphthol solution, indomethacin, and carrageenan (Sigma-Aldrich, India)\u003c/li\u003e\n \u003cli\u003eSodium carbonate (Rankem, India)\u003c/li\u003e\n \u003cli\u003eAcetic acid, acetic anhydride, concentrated sulfuric acid (Rankem, India)\u003c/li\u003e\n \u003cli\u003eAmmonia, Benedict’s reagent, Biuret reagent, diluted hydrochloric acid, ferric chloride, Folin-Ciocalteu phenol reagent, Hager’s reagent, Mayer’s reagent, pyridine, sodium hydroxide solution, sodium nitroprusside solution, and Sudan Red III reagent (Sigma Aldrich Chemicals, India)\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eDPPH (2,2-diphenyl-1-picrylhydrazyl) powder\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eEthanol (analytical grade)\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eMethanolic extract of the sample specimen\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eAscorbic acid (standard antioxidant)\u003c/li\u003e\n\u003c/ul\u003e\n\u003ch3\u003eApparatus and Equipment\u003c/h3\u003e\n\u003ch4\u003eLaboratory Glassware\u003c/h4\u003e\n\u003cp\u003eVarious laboratory glassware items were utilized, including beakers, conical flasks, round bottom flasks, a grinder for plant material, measuring cylinders, porcelain dishes, heating mantles, filter papers, test tubes, stirring glass rods, micro hematocrit tubes, a digital weighing balance, and a water bath.\u003c/p\u003e\n\u003ch4\u003e\u003cstrong\u003eSpecific Instruments\u003c/strong\u003e\u003c/h4\u003e\n\u003cp\u003eSpecific instruments included a Soxhlet extractor with a condenser, a hot air oven, a desiccator, digital calipers, a microscope, a rotavapour, insulin syringe needles and a metal gavage needle.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003ePreparation of Plant Extracts\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe plant leaves were dried in the shade and ground into coarse powder. Approximately 230 g of this powder was placed in a Soxhlet extraction apparatus [8]. Extraction was performed using chloroform, methanol, and water, each at twice the weight of the dried plant powder. Solvents wetted the powder and filled the round bottom flask, which was heated to evaporate the solvent. The vapor passed through the plant material, condensed, and dripped back onto the powder. Extraction continued until the solvent in the siphon lost its color. The extract was then filtered to remove impurities. Following Soxhlet extraction with chloroform and methanol, the remaining sample underwent distilled water extraction using maceration. The sample was soaked in distilled water for three days with periodic agitation and then strained. Chloroform extracted non-polar compounds, methanol extracted polar compounds, and water extracted highly polar compounds.\u0026nbsp;\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003eConcentration of Extracts\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eEach extract underwent concentration using a rotary evaporator with specific conditions: the chloroform extract was concentrated at 62°C under 475 mbar pressure, the methanol extract at 65°C under 335 mbar pressure, and the distilled water extract at 100°C under 72 mbar pressure. After concentration, the extracts were stored in a desiccator containing calcium chloride to eliminate moisture. Finally, the concentrated extracts were transferred into airtight containers and stored at 4°C.\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003ePreliminary Phytochemical Analysis\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eAll extracted samples were subjected to preliminary phytochemical analysis to qualitatively identify various compounds present. The screening aimed to detect alkaloids, flavonoids, terpenoids, phenolic compounds, tannins, glycosides, saponins, fixed oils and fats, carbohydrates, and proteins within the plant extracts [9].\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003eExperimental Design for Rat Paw Edema Study [10]\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eThirty healthy Swiss albino rats weighing between 150-210 g were chosen for the experiment. The rats were randomly divided into five groups, each consisting of six rats. These groups included a positive control, a negative control, and three experimental groups treated with previously extracted plant extracts. The chloroform, methanol, and distilled water extracts were administered orally at a dose of 100 mg/kg to their respective groups. Additionally, standard drug indomethacin and normal saline were administered orally as controls. Figure 2 depicts the oral administration of extracts in rats.\u003c/p\u003e\n\u003cp\u003ePreparations of the extracts were tailored to each rat's weight prior to administration. The chloroform extract was suspended in an emulsion of olive oil, distilled water, and acacia gum (in a ratio of 4:2:1) due to its insolubility in water. The methanol and distilled water extracts were dissolved in distilled water before administration. Table 1 provides an overview of the experimental groups and their respective treatments.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable-1: Description of experimental design groups\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"20.555555555555557%\" valign=\"top\"\u003e\n \u003cp\u003eGroups (n=6)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"40.55555555555556%\" valign=\"top\"\u003e\n \u003cp\u003eExperimental design groups\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"38.888888888888886%\" valign=\"top\"\u003e\n \u003cp\u003eTreatment\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20.555555555555557%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"40.55555555555556%\" valign=\"top\"\u003e\n \u003cp\u003ePositive control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"38.888888888888886%\" valign=\"top\"\u003e\n \u003cp\u003eCarrageenan + 10 mg/kg Indomethacin\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20.555555555555557%\" valign=\"top\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"40.55555555555556%\" valign=\"top\"\u003e\n \u003cp\u003eChloroform\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"38.888888888888886%\" valign=\"top\"\u003e\n \u003cp\u003eCarrageenan + 100 mg/kg Chloroform extract\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20.555555555555557%\" valign=\"top\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"40.55555555555556%\" valign=\"top\"\u003e\n \u003cp\u003eMethanol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"38.888888888888886%\" valign=\"top\"\u003e\n \u003cp\u003eCarrageenan + 100 mg/kg Methanol extract\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20.555555555555557%\" valign=\"top\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"40.55555555555556%\" valign=\"top\"\u003e\n \u003cp\u003eDistilled water\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"38.888888888888886%\" valign=\"top\"\u003e\n \u003cp\u003eCarrageenan + 100 mg/kg Distilled water extract\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20.555555555555557%\" valign=\"top\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"40.55555555555556%\" valign=\"top\"\u003e\n \u003cp\u003eNegative control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"38.888888888888886%\" valign=\"top\"\u003e\n \u003cp\u003eCarrageenan + Normal Saline\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eEach group consisted of 6 rats (n=6), and these treatments were administered to evaluate the anti-inflammatory effects using a carrageenan-induced paw edema model.\u003c/p\u003e\n\u003cp\u003eTable1 provides an overview of the experimental groups in the study, detailing their respective treatments: the positive control received carrageenan injection followed by 10 mg/kg of indomethacin; the chloroform, methanol, and distilled water extract groups received carrageenan injection followed by 100 mg/kg of their respective extracts; and the negative control received carrageenan injection followed by normal saline.\u003c/p\u003e\n\u003cp\u003eSix rats were randomly placed into labeled cages, with each cage representing a different experimental group. Paw thickness was measured for each rat every 60 minutes from 0 to 300 minutes during the experiment. After the experiment, the percentage of edema inhibition was calculated for each set of data.\u003c/p\u003e\n\u003cp\u003eThe anti-inflammatory potential of various extracts of \u003cem\u003eMillettia peguensis\u003c/em\u003e was assessed using the carrageenan-induced hind paw edema model in albino rats. Indomethacin was administered as a standard treatment in the positive control group at a dosage of 10 mg/kg. Each extract was administered at a dose of 100 mg/kg body weight to separate groups of rats, one hour before inducing inflammation by injecting carrageenan into the left hind paw plantar tissue. Carrageenan was prepared by dissolving 1% solution in normal saline to create a 0.1 mL injection. Paw thickness was measured before carrageenan injection and hourly thereafter for five hours using a digital caliper, measuring at the dorsa-plantar axis at the metatarsal level.\u003c/p\u003e\n\u003cp\u003eThe anti-inflammatory activity was quantified as the percentage of edema inhibition in the positive control and extract-treated groups compared to the negative control group. The percentage of edema inhibition was calculated using the formula:\u003c/p\u003e\n\u003cp\u003e\u003cimg src=\"data:image/png;base64,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\" width=\"590\" height=\"50\"\u003e\u003c/p\u003e\n\u003cp\u003eWhere Tt is the mean paw thickness of the treated group rats and Tc is the mean paw thickness of the control group rats at the same time interval.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAdministration of drugs and experimental compounds in rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eProper handling and restraint techniques were employed to minimize stress during procedures. The V-grip method, following [11] gently gripped the rat's head with knuckles on the jaw bone and secured the thorax under both forelimbs. This method, recommended for the dominant hand, supports the rat's lower body or rests it on the handler's chest for comfort, as suggested by Moore [12].\u003c/p\u003e\n\u003cp\u003eOral gavage involved administering substances directly into the stomach using a specialized blunt-ended gavage needle with a ball tip, ensuring precise dosing and rapid absorption compared to dietary methods. Maximum volumes were based on individual rat weights, and needle lengths were adjusted to avoid injury or aspiration. The syringe was prepared with the substance, and the needle surface wiped clean to prevent residue affecting taste.\u003c/p\u003e\n\u003cp\u003eRats were gently restrained using the V-grip method, the needle inserted into the back of the throat along the mouth's left side and roof, and the rat's head tilted back to straighten the esophagus. The needle was advanced until the pre-marked line reached the mouth, with occasional slight twists to pass the epiglottis. The substance was slowly injected into the stomach, and the needle withdrawn carefully before the rat was returned to its cage.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIntraplantar Injection\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe carrageenan solution was injected intraplantarly into rats, a technique blending aspects of subcutaneous and intradermal injections widely used in inflammation, pain, and immunization studies [13]. Inflammation was induced exclusively in the left hind paw with an injection volume not exceeding 0.10 mL per rat. Before injection, the site was sterilized with a 70% isopropyl alcohol swab. A needle size of 25G or smaller delivered 0.1 mL of 1% carrageenan solution in normal saline just beneath the epidermis, at a shallow angle. Post-injection, the needle was gently withdrawn with light pressure on the site to prevent leakage. Firm pressure was briefly applied to confirm successful injection, evidenced by the formation of a distinct bleb at the site. Figure 3 shows the measurement of paw thickness with a digital Vernier caliper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± standard deviation (SD) and analyzed using Statistical Package for the Social Sciences (SPSS) version 23.0. One-way analysis of variance (ANOVA) followed by Tukey's honestly significant difference (HSD) post hoc test was employed for statistical comparisons, with significance set at p \u0026lt; 0.05.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntioxidant Activity of Methanolic Extract of \u003cem\u003eMillettia peguensis\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cem\u003e[14]\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of DPPH Solution:\u0026nbsp;\u003c/strong\u003eDissolved 4 mg of DPPH in 100 mL of ethanol to prepare a 0.1 mM DPPH solution. Protected the solution from light by covering the container with aluminum foil or using an amber bottle.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of Sample Extract Solutions:\u0026nbsp;\u003c/strong\u003ePrepared different concentrations of the methanolic extract of the sample specimen (e.g., 10, 25, 50, 75, 100 µg/mL) by diluting the stock extract with methanol.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of Standard Solution:\u0026nbsp;\u003c/strong\u003ePrepared a standard solution of ascorbic acid (e.g., 10, 25, 50, 75, 100 µg/mL) in ethanol or methanol.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eReaction Mixture Preparation:\u003c/strong\u003e\u003c/p\u003e\n\u003cul type=\"disc\"\u003e\n \u003cli\u003eIn test tubes or a 96-well microplate, 1 mL of DPPH solution was added to 1 mL of each concentration of the sample extract.\u003c/li\u003e\n \u003cli\u003eFor the standard, 1 mL of DPPH solution was added to 1 mL of each concentration of the ascorbic acid solution.\u003c/li\u003e\n \u003cli\u003ePrepared a control by adding 1 mL of DPPH solution to 1 mL of ethanol (without the sample extract or standard).\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003e\u003cstrong\u003eIncubation:\u0026nbsp;\u003c/strong\u003eVortexed the mixtures to ensure thorough mixing. Incubated the mixtures at room temperature in the dark for 30 minutes.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMeasurement of Absorbance:\u0026nbsp;\u003c/strong\u003eMeasured the absorbance of each reaction mixture at 517 nm using a UV-Vis spectrophotometer. The decrease in absorbance indicates the scavenging of DPPH free radicals by the antioxidants present in the sample extract or standard.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCalculation of DPPH Radical Scavenging Activity:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cimg src=\"data:image/png;base64,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\"\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePlotting the Results:\u003c/strong\u003e\u003c/p\u003e\n\u003cul type=\"disc\"\u003e\n \u003cli\u003ePlotted the scavenging activity (%) against the concentrations of the sample extract and standard.\u003c/li\u003e\n \u003cli\u003eDetermined the IC50 value, which was the concentration of the sample required to scavenge 50% of the DPPH radicals.\u003c/li\u003e\n\u003c/ul\u003e"},{"header":"RESULTS AND DISCUSSION","content":"\u003cp\u003e\u003cstrong\u003eExtraction of \u003cem\u003eMillettia peguensis\u003c/em\u003e Leaves\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe extraction of \u003cem\u003eMillettia peguensis\u003c/em\u003e leaves involved the sequential use of three solvents: chloroform, methanol, and distilled water, chosen based on their varying polarities to extract secondary metabolites of interest. Initially, 230 g of dried leaf powder was subjected to extraction using a Soxhlet extractor with chloroform and methanol, followed by maceration with distilled water. The choice of solvent followed a gradient from low to high polarity. The percentage yield of each solvent extraction was calculated using the formula:\u003c/p\u003e\n\u003cp\u003e\u003cimg src=\"data:image/png;base64,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\" width=\"304\" height=\"55\"\u003e\u003c/p\u003e\n\u003cp\u003eThe extracted masses, percentage yields, colors, and consistencies of the \u003cem\u003eMillettia peguensis\u003c/em\u003e leaf extracts are summarized in Table 2.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2: Extraction metrics of \u003cem\u003eMillettia peguensis\u003c/em\u003e Leaves Extracts\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eExtract\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eWeight (g)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.526627218934912%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eYield (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eColor\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eConsistency\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003eChloroform\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003e19.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.526627218934912%\" valign=\"top\"\u003e\n \u003cp\u003e8.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003eGrey-black\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003eSolid\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003eMethanol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003e43.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.526627218934912%\" valign=\"top\"\u003e\n \u003cp\u003e19.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003eGrey-black\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003eSemisolid\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003eWater\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003e12.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.526627218934912%\" valign=\"top\"\u003e\n \u003cp\u003e5.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003eDark brown\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.118343195266274%\" valign=\"top\"\u003e\n \u003cp\u003eSemisolid\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eThis table shows plant extracts using chloroform, methanol, and water, with yields of 8.31%, 19.02%, and 5.49%, respectively. Chloroform and methanol extracts were grey-black, with chloroform being solid and methanol semisolid. The water extract was dark brown and semisolid, reflecting differences in solvent polarity and extracted compounds. \u0026nbsp;\u003c/p\u003e\n\u003ch2\u003ePhytochemical analysis of the extracts:\u0026nbsp;\u003c/h2\u003e\n\u003cp\u003eThe \u003cem\u003eMillettia peguensis\u003c/em\u003e leaf extracts were tested for phytochemicals. Results were shown below. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 3: Results of phytochemical analysis of \u003cem\u003eMillettia peguensis\u003c/em\u003e leaf extracts\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"636\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.9811320754717%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eConstituent\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"34.905660377358494%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTypes of test\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.09433962264151%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eChloroform\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.20754716981132%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMethanol\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.81132075471698%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDistilled water\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.9811320754717%\" valign=\"top\"\u003e\n \u003cp\u003eFlavonoids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"34.905660377358494%\" valign=\"top\"\u003e\n \u003cp\u003eAlkaline reagent test\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.09433962264151%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.20754716981132%\" valign=\"top\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.81132075471698%\" valign=\"top\"\u003e\n \u003cp\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.9811320754717%\" rowspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003eTerpenoids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"34.905660377358494%\" valign=\"top\"\u003e\n \u003cp\u003eSalkowski reaction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.09433962264151%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.20754716981132%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.81132075471698%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"42.04545454545455%\" valign=\"top\"\u003e\n \u003cp\u003eLiebermann-Burchard reaction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.181818181818183%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.909090909090908%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"23.863636363636363%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.9811320754717%\" rowspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003eTannins\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"34.905660377358494%\" valign=\"top\"\u003e\n \u003cp\u003e5% Ferric chloride\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.09433962264151%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.20754716981132%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.81132075471698%\" valign=\"top\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"42.04545454545455%\" valign=\"top\"\u003e\n \u003cp\u003eAcetic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.181818181818183%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.909090909090908%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"23.863636363636363%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.9811320754717%\" rowspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003eAlkaloids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"34.905660377358494%\" valign=\"top\"\u003e\n \u003cp\u003eMayer\u0026rsquo;s test\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.09433962264151%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.20754716981132%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.81132075471698%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"42.04545454545455%\" valign=\"top\"\u003e\n \u003cp\u003eHager\u0026rsquo;s test\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.181818181818183%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.909090909090908%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"23.863636363636363%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.9811320754717%\" rowspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003eGlycosides\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"34.905660377358494%\" valign=\"top\"\u003e\n \u003cp\u003eLegal\u0026rsquo;s test\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.09433962264151%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.20754716981132%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.81132075471698%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"42.04545454545455%\" valign=\"top\"\u003e\n \u003cp\u003eModified Borntrager\u0026rsquo;s test\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.181818181818183%\" valign=\"top\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.909090909090908%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"23.863636363636363%\" valign=\"top\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.9811320754717%\" rowspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003ePhenolic compounds\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"34.905660377358494%\" valign=\"top\"\u003e\n \u003cp\u003eFolin-Ciocalteau Test\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.09433962264151%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.20754716981132%\" valign=\"top\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.81132075471698%\" valign=\"top\"\u003e\n \u003cp\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"42.04545454545455%\" valign=\"top\"\u003e\n \u003cp\u003e5% Ferric chloride\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.181818181818183%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.909090909090908%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"23.863636363636363%\" valign=\"top\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.9811320754717%\" valign=\"top\"\u003e\n \u003cp\u003eSaponins\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"34.905660377358494%\" valign=\"top\"\u003e\n \u003cp\u003eFoam test\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.09433962264151%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.20754716981132%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.81132075471698%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.9811320754717%\" rowspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003eCarbohydrates\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"34.905660377358494%\" valign=\"top\"\u003e\n \u003cp\u003eMolisch\u0026rsquo;s test\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.09433962264151%\" valign=\"top\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.20754716981132%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.81132075471698%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"42.04545454545455%\" valign=\"top\"\u003e\n \u003cp\u003eBenedict\u0026rsquo;s test\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.181818181818183%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.909090909090908%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"23.863636363636363%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.9811320754717%\" rowspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003eProtein\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"34.905660377358494%\" valign=\"top\"\u003e\n \u003cp\u003eBiuret test\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.09433962264151%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.20754716981132%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.81132075471698%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"42.04545454545455%\" valign=\"top\"\u003e\n \u003cp\u003eXanthoprotein test\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.181818181818183%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.909090909090908%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"23.863636363636363%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.9811320754717%\" valign=\"top\"\u003e\n \u003cp\u003eFats and oils\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"34.905660377358494%\" valign=\"top\"\u003e\n \u003cp\u003eSudan red III reagent test\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.09433962264151%\" valign=\"top\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.20754716981132%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.81132075471698%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cul type=\"disc\"\u003e\n \u003cli\u003e- : Not present \u0026nbsp; \u0026nbsp; \u0026nbsp;+ : Present in low concentration \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;++ : Present in high concentration\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003eTable 3 compares phytochemical constituents in an extract using chloroform, methanol, and distilled water. Flavonoids are more pronounced in methanol and water, indicated by strong alkaline reagent reactions. Terpenoids are detected in chloroform and methanol but absent in water. Tannins show higher presence in methanol and water via ferric chloride tests. Alkaloids are solely detected in methanol. Glycosides are minimal, observed only in chloroform with moderate response in Borntrager\u0026rsquo;s test. Phenolic compounds are prominent in methanol and water, notably with Folin-Ciocalteau and ferric chloride tests. Saponins are positive in methanol and water. Carbohydrates are detected in all solvents. Protein, fats, and oils are minimally detected, predominantly in chloroform.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreliminary paw thickness of normal albino rats [15]\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eInitial paw thickness of normal albino rats was measured before inducing edema in 30 randomly grouped rats, divided into five groups with six rats each. Paw thickness was assessed using a calibrated digital Vernier caliper along the dorsa-plantar axis at the metatarsal level, as depicted in Table 4 shows the initial mean paw thickness for each group. Analysis indicated no significant difference in initial paw thickness between the control and experimental groups, confirming uniformity among all groups before inducing edema.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTable 4 presents initial paw thickness measurements (mm) for experimental groups of albino rats: positive control (indomethacin), chloroform extract, methanol extract, distilled water extract, and negative control (normal saline). Each value is mean \u0026plusmn; standard deviation (S.D.) from six rats/group.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4: Initial Paw Thickness in Various Experimental Groups\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"50%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eAlbino Wistar Group\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eInitial paw thickness (mm)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"50%\" valign=\"top\"\u003e\n \u003cp\u003ePositive control, Indomethacin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50%\" valign=\"top\"\u003e\n \u003cp\u003e2.27 \u0026plusmn; 0.19\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"50%\" valign=\"top\"\u003e\n \u003cp\u003eChloroform\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50%\" valign=\"top\"\u003e\n \u003cp\u003e2.50 \u0026plusmn; 0.48\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"50%\" valign=\"top\"\u003e\n \u003cp\u003eMethanol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50%\" valign=\"top\"\u003e\n \u003cp\u003e2.48 \u0026plusmn; 0.32\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"50%\" valign=\"top\"\u003e\n \u003cp\u003eDistilled water\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50%\" valign=\"top\"\u003e\n \u003cp\u003e2.26 \u0026plusmn; 0.12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"50%\" valign=\"top\"\u003e\n \u003cp\u003eNegative control, normal saline\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50%\" valign=\"top\"\u003e\n \u003cp\u003e2.46 \u0026plusmn; 0.19\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eStatistical analysis (p \u0026gt; 0.05) indicated no significant differences in initial paw thickness, ensuring uniformity at the study\u0026apos;s outset. These measurements serve as a baseline for comparing subsequent paw thickness post-edema induction, facilitating evaluation of treatment effects on inflammation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEvaluation of anti-inflammatory activity of various extracts from Millettia peguensis leaves on rat paw edema induced by carrageenan\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study assessed anti-inflammatory effects of \u003cem\u003eMillettia peguensis\u003c/em\u003e leaf extracts on carrageenan-induced paw edema in rats. Extracts in chloroform, methanol, and water, alongside indomethacin (standard drug), were orally administered at 100 mg/kg and 10 mg/kg doses, one-hour pre-injection. Paw thickness was measured hourly post-injection over five hours to evaluate inflammation impact. Results, including paw thickness and percentage inhibition of inflammation, are summarized in Table 5. ANOVA with Tukey\u0026rsquo;s HSD test indicated significant differences (p \u0026lt; 0.05) in paw thickness among treatment groups. \u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 5\u003c/strong\u003e Data shows that extracts of \u003cem\u003eMillettia peguensis\u003c/em\u003e and indomethacin reduce rat paw thickness and edema at different times.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"10.576923076923077%\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroup\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"89.42307692307692%\" colspan=\"5\"\u003e\n \u003cp\u003e\u003cstrong\u003eTime interval\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003csup\u003est\u003c/sup\u003e hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003csup\u003end\u003c/sup\u003e hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003csup\u003erd\u003c/sup\u003e hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4\u003csup\u003eth\u003c/sup\u003e hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5\u003csup\u003eth\u003c/sup\u003e hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"10.543130990415335%\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e3.42\u0026nbsp;\u0026plusmn; 0.46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e3.68 \u0026plusmn; 0.47 \u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e3.46 \u0026plusmn; 0.51 \u003csup\u003eb, e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e3.12 \u0026plusmn; 0.45 \u003csup\u003eb, c, e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e2.94 \u0026plusmn; 0.50 \u003csup\u003eb, e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(14.26)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(15.20)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(22.99)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(30.33)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(33.61)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"10.543130990415335%\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e4.12\u0026nbsp;\u0026plusmn; 0.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e4.37\u0026nbsp;\u0026plusmn; 0.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e4.35 \u0026plusmn; 0.30 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e4.07 \u0026plusmn; 0.27 \u003csup\u003ea, e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e3.78\u0026nbsp;\u0026plusmn; 0.30\u003csup\u003e\u0026nbsp; a, e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(1.39)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(3.17)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(8.74)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(16.08)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(21.56)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"10.543130990415335%\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e4.04\u0026nbsp;\u0026plusmn; 0.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e4.19\u0026nbsp;\u0026plusmn; 0.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e4.24\u0026nbsp;\u0026plusmn; 0.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e3.99\u0026nbsp;\u0026plusmn; 0.40\u003csup\u003e\u0026nbsp;a, e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e3.58 \u0026plusmn; 0.35 \u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(2.95)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(6.32)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(10.61)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(17.41)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(24.55)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"10.543130990415335%\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e4\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e4.01\u0026nbsp;\u0026plusmn; 0.64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e4.09\u0026nbsp;\u0026plusmn; 0.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e3.74\u0026nbsp;\u0026plusmn; 0.81\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e3.60 \u0026nbsp;\u0026plusmn; 0.83\u003csup\u003e\u0026nbsp;e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e3.31 \u0026plusmn; 0.74 \u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(3.17)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(8.09)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(18.62)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(23.32)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(28.37)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"10.543130990415335%\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e4.19\u0026nbsp;\u0026plusmn; 0.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e4.55 \u0026nbsp;\u0026plusmn; 0.27\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e4.87 \u0026plusmn; 0.16 \u003csup\u003ea, d\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e5.08\u0026nbsp;\u0026plusmn; 0.21\u003csup\u003ea, b, c, d\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.891373801916934%\"\u003e\n \u003cp\u003e5.16 \u0026plusmn; 0.28 \u003csup\u003ea, b, c, d\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(0.00)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(0.00)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(0.00)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(0.00)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e(0.00)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eNote: Data represent the mean \u0026plusmn; SD of 6 animals in each group. Values within parenthesis indicate percentage of inhibition (%). \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n\u003cp\u003ea: Significant compared to group 1 (indomethacin; p\u0026lt;0.05). b: Significant compared to group 2 (chloroform extract; p\u0026lt;0.05). c: Significant compared to group 3 (methanol extract; p\u0026lt;0.05). d: Significant compared to group 4 (distilled water extract; p\u0026lt;0.05). e: Significant compared to group 5 (negative control; p\u0026lt;0.05).\u003c/p\u003e\n\u003cp\u003eIn rats, intraplantar carrageenan injection caused paw volume to increase over time, peaking at 5.66 \u0026plusmn; 0.31 mm by the fifth hour post-injection. Initially, chloroform and methanol extracts did not significantly affect paw swelling until after the fourth hour. Indomethacin and distilled water extract demonstrated significant anti-inflammatory effects starting from the second and third hour post-injection, respectively.\u003c/p\u003e\n\u003cp\u003eIndomethacin treatment exhibited the highest paw edema inhibition (33.61%) compared to other treatments, significantly reducing swelling starting from the second hour (peak:\u0026nbsp;3.68\u0026nbsp;\u0026plusmn; 0.47\u0026nbsp;mm) compared to the negative control group.\u003c/p\u003e\n\u003cp\u003eBy the third hour post-injection, chloroform extract, distilled water extract, and indomethacin treatment groups showed decreasing paw swelling. Distilled water extract significantly reduced swelling compared to the control group (p\u0026lt;0.05), while chloroform extract (4.35\u0026nbsp;\u0026plusmn; 0.30\u0026nbsp;mm) did not show significant effects at the same time point.\u003c/p\u003e\n\u003cp\u003eThe paw thickness in the methanol group decreased from\u0026nbsp;4.24\u0026nbsp;\u0026plusmn; 0.45mm at the third hour post-carrageenan injection to\u0026nbsp;3.99\u0026nbsp;\u0026plusmn; 0.40\u003csup\u003e\u0026nbsp;\u003c/sup\u003emm at the fourth hour. Paw thickness continued to decrease in other groups, with indomethacin treatment showing significantly greater reduction compared to chloroform and methanol extracts.\u003c/p\u003e\n\u003cp\u003eDistilled water extract exhibited the highest inhibition percentage (28.37%), followed by methanol extract (24.55%), both significantly different from the negative control group. Chloroform extract also showed significant edema suppression (21.56%) compared to the negative control, albeit less effective than indomethacin.\u003c/p\u003e\n\u003cp\u003ePhytochemical analysis revealed that methanol extract contained more active constituents than distilled water extract, specifically alkaloids and terpenoids. However, distilled water extract showed superior edema inhibition, possibly due to higher concentrations of constituents detected, including flavonoids, tannins, and phenolic compounds. Chloroform extract exhibited the lowest variety of active constituents and demonstrated relatively weaker edema inhibition compared to distilled water and methanol extracts. Therefore, at 100 mg/kg, distilled water extract of \u003cem\u003eMillettia peguensis\u003c/em\u003e reduces carrageenan-induced edema similarly to the potent anti-inflammatory drug indomethacin in a time-dependent manner.\u003c/p\u003e\n\u003cp\u003eThe swelling level was measured by recording the increase in paw thickness at different times, shown in Table 6, with data presented as the swelling level \u0026plusmn; standard deviation.\u0026nbsp;\u003c/p\u003e\n\u003ch4\u003e\u003cstrong\u003eComparative Analysis of Swelling Degrees Over Five Hours\u003c/strong\u003e\u003c/h4\u003e\n\u003cp\u003eTable 6 presents a comparative analysis of the degree of swelling (mm) in various groups over a five-hour period. Each group, including positive and negative controls, and chloroform, methanol, and distilled water-treated samples, were evaluated for swelling at hourly intervals. The values are expressed as means \u0026plusmn; standard deviation, allowing for precise assessment of each treatment\u0026apos;s anti-inflammatory potential.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 6: Level of swelling at different time intervals\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.991150442477878%\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroup\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"83.00884955752213%\" colspan=\"5\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDegree of swelling (mm)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003csup\u003est\u003c/sup\u003e hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003csup\u003end\u003c/sup\u003e hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003csup\u003erd\u003c/sup\u003e hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4\u003csup\u003eth\u003c/sup\u003e hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5\u003csup\u003eth\u003c/sup\u003e hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.96113074204947%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePositive control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.33 \u0026plusmn;0.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.58 \u0026plusmn;0.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.36 \u0026plusmn;0.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.02 \u0026plusmn;0.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e0.84 \u0026plusmn;0.44\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.96113074204947%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eChloroform\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.78\u0026nbsp;\u0026plusmn;0.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e2.04\u0026nbsp;\u0026plusmn;0.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e2.02\u0026nbsp;\u0026plusmn;0.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.74\u0026nbsp;\u0026plusmn;0.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.45\u0026nbsp;\u0026plusmn;0.20\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.96113074204947%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMethanol\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.73\u0026nbsp;\u0026plusmn;0.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.88\u0026nbsp;\u0026plusmn;0.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.93\u0026nbsp;\u0026plusmn;0.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.68\u0026nbsp;\u0026plusmn;0.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.27\u0026nbsp;\u0026plusmn;0.44\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.96113074204947%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDistilled water\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.92\u0026nbsp;\u0026plusmn;0.68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e2.00\u0026nbsp;\u0026plusmn;0.77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.65\u0026nbsp;\u0026plusmn;0.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.51\u0026nbsp;\u0026plusmn;0.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.22\u0026nbsp;\u0026plusmn;0.79\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.96113074204947%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eNegative control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e1.90\u0026nbsp;\u0026plusmn;0.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e2.26\u0026nbsp;\u0026plusmn;0.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e2.58\u0026nbsp;\u0026plusmn;0.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e2.79\u0026nbsp;\u0026plusmn;0.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.607773851590107%\" valign=\"top\"\u003e\n \u003cp\u003e2.87\u0026nbsp;\u0026plusmn;0.38\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003ch4\u003e\u003cstrong\u003eTreatment-specific Observations and Temporal Trends\u003c/strong\u003e\u003c/h4\u003e\n\u003cp\u003eThe positive control group shows a gradual reduction in swelling from 1.33 \u0026plusmn; 0.34 mm at the first hour to 0.84 \u0026plusmn; 0.44 mm at the fifth hour, indicating effective anti-inflammatory activity. Conversely, the negative control group exhibits a continuous increase in swelling, from 1.90 \u0026plusmn; 0.36 mm at the first hour to 2.87 \u0026plusmn; 0.38 mm by the fifth hour, highlighting the baseline inflammatory response without treatment.\u003c/p\u003e\n\u003cp\u003eIn the chloroform-treated group, swelling initially increases from 1.78 \u0026plusmn; 0.30 mm, peaking at 2.04 \u0026plusmn; 0.28 mm in the second hour, and then declines to 1.45 \u0026plusmn; 0.20 mm by the fifth hour, suggesting a moderate anti-inflammatory effect. The methanol-treated group shows similar trends, with initial swelling of 1.73 \u0026plusmn; 0.53 mm, peaking at 1.93 \u0026plusmn; 0.50 mm at the third hour, and reducing to 1.27 \u0026plusmn; 0.44 mm at the fifth hour.\u003c/p\u003e\n\u003ch4\u003e\u003cstrong\u003eDistilled Water Treatment and Comparative Efficacy\u003c/strong\u003e\u003c/h4\u003e\n\u003cp\u003eThe distilled water-treated group shows initial swelling of 1.92 \u0026plusmn; 0.68 mm, peaking at 2.00 \u0026plusmn; 0.77 mm at the second hour, and declining to 1.22 \u0026plusmn; 0.79 mm by the fifth hour. This pattern indicates that distilled water has moderate anti-inflammatory properties, comparable to methanol and slightly better than chloroform.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntioxidant activity of methanolic extract of \u003cem\u003eMillettia peguensis\u003c/em\u003e by DPPH method\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe table-7 presents the in vitro antioxidant activity of different concentrations of an extract using the DPPH method. The % inhibition of DPPH free radicals was measured at concentrations ranging from 5 to 60 \u0026mu;l/ml. As the concentration increases, the % inhibition also increases, indicating higher antioxidant activity. At the lowest concentration (5 \u0026mu;l/ml), the % inhibition values are 16.18%, 20.23%, and 29.84% for the different extracts tested. As the concentration increases to 60 \u0026mu;l/ml, the % inhibition reaches 81.07%, 89.22%, and 99.74%, respectively. This trend demonstrates a dose-dependent increase in antioxidant activity for all extracts tested, with the highest concentration showing the greatest inhibition of DPPH radicals which were shown in Figure 4.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 7: Data of\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eAntioxidant activity of Various extracts of\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eMillettia peguensis\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"19.387755102040817%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentration\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e[\u0026mu;l/ml])\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e% Inhibition by Chloroform extract\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.489795918367346%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e% Inhibition by Methanolic extract\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"30.612244897959183%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e% Inhibition by Distilled water extract\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"19.387755102040817%\" valign=\"top\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\" valign=\"top\"\u003e\n \u003cp\u003e16.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.489795918367346%\" valign=\"top\"\u003e\n \u003cp\u003e20.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"30.612244897959183%\" valign=\"top\"\u003e\n \u003cp\u003e29.84\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"19.387755102040817%\" valign=\"top\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\" valign=\"top\"\u003e\n \u003cp\u003e22.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.489795918367346%\" valign=\"top\"\u003e\n \u003cp\u003e33.46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"30.612244897959183%\" valign=\"top\"\u003e\n \u003cp\u003e42.10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"19.387755102040817%\" valign=\"top\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\" valign=\"top\"\u003e\n \u003cp\u003e32.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.489795918367346%\" valign=\"top\"\u003e\n \u003cp\u003e36.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"30.612244897959183%\" valign=\"top\"\u003e\n \u003cp\u003e50.16\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"19.387755102040817%\" valign=\"top\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\" valign=\"top\"\u003e\n \u003cp\u003e42.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.489795918367346%\" valign=\"top\"\u003e\n \u003cp\u003e43.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"30.612244897959183%\" valign=\"top\"\u003e\n \u003cp\u003e59.35\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"19.387755102040817%\" valign=\"top\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\" valign=\"top\"\u003e\n \u003cp\u003e42.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.489795918367346%\" valign=\"top\"\u003e\n \u003cp\u003e45.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"30.612244897959183%\" valign=\"top\"\u003e\n \u003cp\u003e64.35\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"19.387755102040817%\" valign=\"top\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\" valign=\"top\"\u003e\n \u003cp\u003e52.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.489795918367346%\" valign=\"top\"\u003e\n \u003cp\u003e57.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"30.612244897959183%\" valign=\"top\"\u003e\n \u003cp\u003e74.84\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"19.387755102040817%\" valign=\"top\"\u003e\n \u003cp\u003e35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\" valign=\"top\"\u003e\n \u003cp\u003e56.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.489795918367346%\" valign=\"top\"\u003e\n \u003cp\u003e67.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"30.612244897959183%\" valign=\"top\"\u003e\n \u003cp\u003e76.29\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"19.387755102040817%\" valign=\"top\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\" valign=\"top\"\u003e\n \u003cp\u003e60.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.489795918367346%\" valign=\"top\"\u003e\n \u003cp\u003e73.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"30.612244897959183%\" valign=\"top\"\u003e\n \u003cp\u003e83.39\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"19.387755102040817%\" valign=\"top\"\u003e\n \u003cp\u003e45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\" valign=\"top\"\u003e\n \u003cp\u003e66.95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.489795918367346%\" valign=\"top\"\u003e\n \u003cp\u003e74.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"30.612244897959183%\" valign=\"top\"\u003e\n \u003cp\u003e87.90\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"19.387755102040817%\" valign=\"top\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\" valign=\"top\"\u003e\n \u003cp\u003e72.98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.489795918367346%\" valign=\"top\"\u003e\n \u003cp\u003e83.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"30.612244897959183%\" valign=\"top\"\u003e\n \u003cp\u003e91.94\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"19.387755102040817%\" valign=\"top\"\u003e\n \u003cp\u003e55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\" valign=\"top\"\u003e\n \u003cp\u003e72.46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.489795918367346%\" valign=\"top\"\u003e\n \u003cp\u003e87.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"30.612244897959183%\" valign=\"top\"\u003e\n \u003cp\u003e97.42\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"19.387755102040817%\" valign=\"top\"\u003e\n \u003cp\u003e60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\" valign=\"top\"\u003e\n \u003cp\u003e81.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.489795918367346%\" valign=\"top\"\u003e\n \u003cp\u003e89.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"30.612244897959183%\" valign=\"top\"\u003e\n \u003cp\u003e99.74\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn conclusion, \u003cem\u003eMillettia peguensis\u003c/em\u003e extracts effectively reduced inflammation in carrageenan-induced rat paw swelling. The extracts, especially distilled water, show significant anti-swelling effects comparable to indomethacin. Phytochemical analysis revealed flavonoids, saponins, phenolic compounds, tannins, and terpenoids, suggesting potential benefits for managing inflammation-related conditions like fever and pain. All extracts demonstrated increasing % inhibition of DPPH radicals with higher concentrations. Methanolic and distilled water extracts consistently show higher inhibition rates compared to chloroform extract, particularly notable at higher concentrations (50-60 \u0026mu;l/ml), suggesting their stronger antioxidant potential. Future research should explore the mechanisms underlying \u003cem\u003eMillettia peguensis\u0026apos;s\u003c/em\u003e anti-inflammatory actions. \u0026nbsp;Continued studies and clinical trials are needed to validate their effectiveness in inflammation reduction, highlighting natural phytoconstituents as promising molecules for drug development. \u0026nbsp;\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eBoth authors, Dr. Nemala Appala Raju and Dr. Anitha Nandagopal, contributed equally to this work. Dr. Nemala Appala Raju and Dr. Anitha Nandagopal were involved in the conceptualization, methodology, formal analysis, and investigation of the research. They jointly handled resources and data curation. The original draft was written by both authors, and both contributed to the review and editing process. Visualization of the results was performed by Dr. Nemala Appala Raju and Dr. Anitha Nandagopal. Supervision and project administration were shared responsibilities between the authors. Both authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe datasets used and/or analysed during the current study available from the corresponding author on reasonable request\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eManikandan G, Vimala Rani A. GC-MS analysis of phytochemical constituents in the petroleum ether leaf extracts of \u003cem\u003eMillettia peguensis\u003c/em\u003e. Int Res J Pharm. 2017; 8 (9):144\u0026ndash;50.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePackiyalakshmi D, Athilakshmi P, Gayathri S, Karthiga P, Thiri Bhuvaneswari R, Manikandan G. Antimicrobial potential of different solvents leaf extract of \u003cem\u003eMillettia peguensis\u003c/em\u003e against selected pathogens. Pharma Inno J. 2017; 8(1): 109\u0026ndash;24.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGanapaty S, Pushpalata V, Gella JB, Naidu KC, Peter WG. Waterman flavonoids from \u003cem\u003eMillettia peguensis\u003c/em\u003e Ali (Fabaceae) Biochem Syst Ecol. 1998; 26(1): 125\u0026ndash;26.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNarkhede CP, Koli SH, Suryawanshi RK, Patil CD, Borase HP, Patil SV. (2016). Potentiation of \u003cem\u003eBacillus thuringiensis\u003c/em\u003e by using some natural products: Novel preparations against dengue vector Aedis aegypti larvae. Indian J Nat Prod Reso. 2016; 7(3): 229\u0026thinsp;\u0026ndash;\u0026thinsp;33.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRao AS, Yadav SS, Singh P, Nandal A, Singh N, Ganaie SA, Yadav N, Kumar R, Bhandoria MS, Bansal P. A comprehensive review on ethnomedicine, phytochemistry, pharmacology, and toxicity of Tephrosia purpurea (L.) Pers. 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Hock immunization: A humane alternative to mouse footpad injections. J Immu Meth. 2007, 328(1\u0026ndash;2): 204\u0026ndash;14.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDontha S. A Review on antioxidant methods. Asian J Pharm Cli Res. 2016; 9(8):14\u0026ndash;32.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eModi F, Varia R, Patel J. Anti-inflammatory activity of ellagic acid in carrageenan-induced paw edema in rats. Haryana Vet. 2023; 62(SI-2): 18\u0026ndash;21.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Millettia peguensis, anti-inflammatory activity, carrageenan-induced paw edema, indomethacin, DPPH","lastPublishedDoi":"10.21203/rs.3.rs-4748245/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4748245/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e \u003cb\u003eMillettia peguensis\u003c/b\u003e is a plant native to Southeast Asia belonging to the Fabaceae family. Leaf extracts were prepared using chloroform, methanol, and water. The carrageenan-induced paw edema model in rats was employed to assess anti-inflammatory activity. Paw thickness was measured over time to quantify edema. All extracts exhibited a reduction in paw edema compared to the control group. The water extract demonstrated a significant reduction at the third hour post-injection. Both methanol and chloroform extracts displayed significant inhibition by the fourth hour, but their efficacy was lower than the positive control group treated with indomethacin. By the fifth hour, both water and methanol extracts produced results comparable to indomethacin. The study assessed DPPH antioxidant activity of extracts from using chloroform, methanol, and distilled water. Methanol and distilled water extracts showed higher antioxidant activity compared to chloroform extract, suggesting the significance of polar compounds. The findings highlight the impact of solvent choice on extracting bioactive antioxidants. Phytochemical analysis suggested the presence of bioactive compounds such as flavonoids, tannins, saponins, and phenolic compounds. Methanol and water extracts showed higher antioxidant activity due to the presence of phenolic compounds, which may be responsible for the observed anti-inflammatory activity.\u003c/p\u003e","manuscriptTitle":"Phytochemical Analysis and Evaluation of the Anti-Inflammatory and Antioxidant Activity of Diverse Extracts of Millettia peguensis Leaves","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-21 11:12:44","doi":"10.21203/rs.3.rs-4748245/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":"a69b72a0-1f96-4dd0-bbec-f3df884c3f3b","owner":[],"postedDate":"August 21st, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-12-20T02:53:26+00:00","versionOfRecord":[],"versionCreatedAt":"2024-08-21 11:12:44","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4748245","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4748245","identity":"rs-4748245","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}