Chemical, Insecticidal, Antioxidant and Phytochemical Assessment of Thorn Apple (Datura metel L.) leaf extract from Pakistan's Subtropical Climate

Chemical, Insecticidal, Antioxidant and Phytochemical Assessment of Thorn Apple (Datura metel L.) leaf extract from Pakistan's Subtropical Climate | 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 Chemical, Insecticidal, Antioxidant and Phytochemical Assessment of Thorn Apple (Datura metel L.) leaf extract from Pakistan's Subtropical Climate Abdul Mateen, Ansar Javeed, Adnan Arshad, Amer Rasul, Shams Ur Rehman, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6110776/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 Datura metel L. , a plant belonging to the solanaceae family, is renowned for its medicinal, agrochemical and poisonous properties. This research assessed the chemical makeup, antioxidant properties, and phytochemical content of D. metel leaf extracts by employing different solvents: ethanol (EtOH), ethyl acetate (EA), dichloromethane (DCM), n-hexane (n-Hx), and distilled water (DW). Chemical profile was assessed by Gas Chromatography-Mass Spectrometry (GC-MS) assay whereas; antioxidant activity was determined by using 1, 1-diphenyl-2-picrylhydrazyl (DPPH) assay. While, Insecticidal activity against Brevicoryne brassicae was evaluated via aphid dip and leaf dip method. On the other hand, quantitative and qualitative phytochemical analysis was carried out by Folin–Ciocalteu reagent method and aluminum chloride colorimetric method, respectively. GC-MS identified four major compounds as: 11H-Pyrido [3’,2’:4,5] imidazo [2,1-b] [1,3] benzothiazin-11-one (29.76%), 1,3-dimethyl Benzene (18.35%), Didodecyl phthalate (11.73%), and 1,3,5-Trimethylbenzene (10.09%), along with six minor compounds. Qualitative phytochemical analysis of plant extract discovered alkaloids, flavonoids, phenols, glycosides, and other substances. Whereas, quantitative tests revealed a high concentration of phenols (75.03 GAEg − 1 ) in distilled water extracts and increased levels of flavonoids in ethyl acetate (99.33 QEg − 1 ). All samples showed significant antioxidant properties, with DPPH inhibition levels ranging from 63.91–81.59%. Similarly, D. metel displayed significant mortality of 81.66% and 73.33% at 48 h exposure at 200 mgmL − 1 via aphid dip and leaf dip assay, respectively. These findings underline the potential for utilizing D. metel leaves as a valuable source of bioactive compounds with significant medicinal advantages. Datura metel GC-MS Phytochemicals DPPH Chemical analysis Insecticide Figures Figure 1 Figure 2 Figure 3 1. Introduction Plants play a vital role in the industrial agrochemical and pharmaceutical sectors, as they contain a wide range of phytochemicals that are effective in treating various diseases. They are the primary sources used in alternative medicine for combating a broad spectrum of ailments. Many traditional medicines come from plants, and around 80% of people worldwide use herbal remedies for maintaining good health. Various plant components such as leaves, flowers, roots, stems, bark, rhizomes, bulbs, and seeds are frequently utilized in making herbal goods (Matotoka & Masoko, 2018 ; Pliego et al., 2022 ). However, the use of medicinal plants varies across regions, influenced by factors such as environmental conditions, cultural practices, historical context, individual beliefs, and philosophical perspectives (Ahmad et al., 2010 ). Medicinal plants are indispensable resources for both traditional and modern medicine, with their usage made by regional, cultural, and environmental factors. Datura metel commonly known as Thorn Apple, is famous for its medicinal properties and its potential use in the agrochemical industry. It is widely cultivated across Asia, Africa, England, India, and various tropical and subtropical regions (Gaire & Subedi, 2013 ). However, in Asia, it typically thrives in wild habitats (Bhattacharjee et al., 2004 ). The medicinal properties of Datura metel are attributed to its bioactive compounds, which have proven effective in treating various conditions, including asthma and bronchitis. Although the plant has narcotic effects, it also offers specific health benefits that make it a valuable a medicinal resource. These benefits include antidiabetic, antimicrobial, anti-inflammatory, anti-asthmatic, antioxidant, insecticidal, cytotoxic, pain-relieving, neurological and, wound healing effects (Alam et al., 2021 ; Shah et al., 2013 ). The plant’s various parts, especially the leaves, contain bioactive compounds, prompting investigations into its chemical composition, antioxidant and phytoconstituents (Rahaman et al., 2023 ). Datura metel 's diverse medicinal properties and bioactive compounds make it a valuable resource for both the pharmaceutical and agrochemical industries. In the agrochemical industry, Datura is recognized for its larvicidal ability to eliminate larvae of the red flour beetle ( Tribolium castaneum ), its effectiveness as a mosquito repellent, and its use in treating animal bites, including snake bites, by providing pain relief (Devi et al., 2011 ; Meselhy, 2012 ). It contains substantial amounts of carbohydrates, moisture, fats, ash, proteins, and crude fiber. Additionally, it is rich in important phytochemicals, including phenolic compounds, alkaloids, flavonoids, cardiac glycosides and, tannins (Bagewadi et al., 2019 ; Fatima et al., 2015 ). Moreover, the significant role of secondary metabolites from the plant kingdom in various biological processes is well established (Divekar et al., 2022 ). Therefore, identifying these valuable phytoconstituents is crucial for understanding their bioactive properties (Xiao & Bai, 2019 ). Thus, rich phytochemical profile and bioactive properties of this plant make it a valuable resource for both agricultural and medicinal applications. Pharmacologically consumption of any part of the Datura plant can cause severe anticholinergic effects, potentially leading to toxicity. While the entire plant contains toxic properties, the seeds are especially harmful. Notably, neither drying nor boiling the plant eliminates its toxic characteristics (Setshogo, 2015 ). Despite its toxicity, D. metel also possesses therapeutic applications, making it a subject of extensive research (Naik et al., 2018 ; Xu et al., 2018 ). This study highlights D. metel as a promising source of bioactive compounds with significant biological properties. By evaluating the chemical composition and antioxidant activity across various extraction methods, valuable insights into the plant's therapeutic potential were gained. The identification of key phytochemicals and their concentrations enhances our understanding of D. metel 's medicinal applications. Given its traditional use in herbal medicine, these findings may guide future research into its role in developing natural remedies and therapeutic agents, underscoring its importance in pharmacological research for disease prevention and treatment. 2. Material and methods 2.1. Collection and authentication of samples During June-July 2024, D. metel leaves were collected from Punjab, Pakistan at the coordinates 32.66843°N and 74.17298°E. Specimens were collected from the identical mature, fully blossoming plant, assuring that only healthy leaves were chosen. This area has a humid subtropical climate in accordance with the Koppen climate classification, which includes four separate seasons. Temperatures in this region fluctuate between 42°F and 103°F annually. The plant species was scientifically verified by Dr. Dilbar Hussain at Ayub Agriculture Research Institute (AARI) Entomological Research Institute Faisalabad Pakistan. 2.2. Extraction procedure The samples were left at room temperature to avoid photo-oxidation until fully dried and were subsequently pulverized into a fine powder with an electric blender. Various solvents such as ethanol (EtOH), ethyl acetate (EA), dichloromethane (DCM), n-hexane (n-Hx), and distilled water (DW) were used to extract the dried powder for 72 hours using a cold extraction method at room temperature. In the extraction procedure, 10g of every dried sample was separately extracted with 100 mL of each solvent, keeping a ratio of 1:10 between sample and solvent (Li Ping et al., 2014 ) stated. The extraction procedure was repeated on three times. The contents extracted were sieved through Whatman filter paper No. 1 and condensed with a rotary evaporator (model R-210 BUCHI Labortechnik AG, CH-9230 Flawil 1/Switzerland) to decrease the volume. After allowing the solvents to completely evaporate in a fume hood for 24 hours, the quantity produced was determined using the Equation (Eq. 1). The samples collected were kept in glass-stoppered vials at 4°C for future phytochemical analysis and evaluation of antioxidant potential (Mueed et al., 2023 ). \(\:\varvec{Y}\varvec{i}\varvec{e}\varvec{l}\varvec{d}\:\left(\varvec{\%}\right)=\frac{\:\mathbf{W}\mathbf{e}\mathbf{i}\mathbf{g}\mathbf{h}\mathbf{t}\:\mathbf{o}\mathbf{f}\:\mathbf{t}\mathbf{h}\mathbf{e}\:\mathbf{e}\mathbf{x}\mathbf{t}\mathbf{r}\mathbf{a}\mathbf{c}\mathbf{t}\:\left(\mathbf{g}\right)}{\:\mathbf{W}\mathbf{e}\mathbf{i}\mathbf{g}\mathbf{h}\mathbf{t}\:\mathbf{o}\mathbf{f}\:\mathbf{t}\mathbf{h}\mathbf{e}\:\mathbf{d}\mathbf{r}\mathbf{i}\mathbf{e}\mathbf{d}\:\mathbf{s}\mathbf{a}\mathbf{m}\mathbf{p}\mathbf{l}\mathbf{e}\:\left(\mathbf{g}\right)\:}\times\:100\varvec{\%}\) (Eq. 1) 2.3 Analysis of chemical compounds GC-MS was performed on the solvent extract of D. metel leaves using an Agilent 6890-5973N system to analyze the chemical compounds. The gas chromatograph was set up using an HP1 capillary column (Model Number: TG-5MS; 30 m × 250 µm × 0.25 µm) and run with a flow rate of 1.0 mL/min. The oven started at 70°C for 0 min, increasing at 3°C/min until reaching 220°C in 10 min. The temperature at the inlet is 250°C, divided at a ratio of 10:1. Included in the additional thermal parameters were a thermal auxiliary temperature of 285°C and a mass spectrometry (MS) scan range from 35 to 520 automatic mass unit (AMU). The MS source was maintained at a temperature of 230°C, with the quadruple temperature held constant at 150°C. Helium served as the carrier gas, flowing at a rate of 1.0mL/min, while HPLC-grade methanol was utilized as the reference sample. The components in the sample were confirmed and authenticated by cross-referencing them with information from the National Institute of Standards and Technology (NIST) Mass Spectral Database (Sparkman, 2005 ). 2.4 Insecticidal activity Insecticidal activity of the D. metel extract was evaluated using two different bioassays like standard leaf dip and individual dip and methods. 2.4.1 Leaf dip assay Using acetone as solvent, already prepared extracts were serially concentrated in Tween-20 (1%) solution at 12.5, 25, 50, 100, and 500 µgmL − 1 . Adult wingless aphids were collected from maize plants that had not been treated with insecticides. In short, fresh maize leaf discs were measuring 5 cm in diameter were cut off, dipped in the corresponding concentration for 10 seconds, and then allowed to dry at room temperature for 5 minutes. A fine-haired aphid was used to carefully release 10 adults, healthy aphids onto the leaf disc in the Petri dish and then incubated for 48 h at 65% relative humidity (R.H), 25˚C temperature, and with a 16:8 (light: dark) photoperiod. 2.4.2 Aphid dip assay In a similar manner, for the aphid dip method10 adult, healthy aphids were carefully dipped in the corresponding concentrations for 5 s before being released on extract free cabbage leaves. The same adjustments were made to the incubation period, R.H., temperature, and photoperiod as with the leaf dip method. The positive control, Imidacloprid 25% WP, was used at a rate of 0.0025 mLmL − 1 in conjunction with control (CK) using a 1% Tween-20 solution. A binocular microscope was used to collect mortality data at 12, 24, and 48 h, and needle probing was used to observe the aphids' reaction. Five duplicates of each treatment were used. 2.5 Assessment of DPPH radical scavenging assay The method proposed by Yu et al., ( 2003 ) was followed by applying DPPH radical scavenging assay in a 1% Tween 20 solution to measure antioxidant activity in the solvent extract of D. metel leaves. Roughly 0.5 mL of the solvent extract was mixed with 3.5 mL of a recently prepared methanol solution containing DPPH (0.004 g/100 mL) in pure methanol of grade HPLC. Following this, 0.25 mL of the extract solution, which had been prepared individually in methanol for each solvent extract, was combined with the freshly made 3.5 mL DPPH solution. The mixture was vigorously shaken to ensure thorough mixing, followed by incubation in darkness at 28°C for 30 min (Roghini & Vijayalakshmi, 2018 ). Following the incubation period, the combination was moved into a 96-well ELISA microplate for absorbance measurement at 517 nm with a photometric plate reader (Spectra-Max 190, Molecular Devices, Shanghai China). Methanol was used as the control group, whereas a combination of DPPH and methanol was used as the experimental group. Eq. 2 was used to calculate the percentage (inhibition) of the DPPH solution by measuring the decrease in absorbance. Higher free radical scavenging activity is reflected by lower absorbance values, as there is an inverse correlation between absorption and radical scavenging activity (Zhao et al., 2008 ). The calculation of the DPPH inhibition percentage was performed with Eq. 2. Where: A blank = (control absorbance); A Sample = (samples absorbance). 2.6 Phytochemical analysis 2.6.1 Qualitative phytochemical assessment Standard procedures were followed to identify different compounds present in the solvent extract of D. metel leaves through qualitative analysis of phytochemicals. These comprised alkaloids (Nortjie et al., 2022 ), anthraquinones (Roghini & Vijayalakshmi, 2018 ), betacyanins (Ali et al., 2018 ), coumarins (Jayapriya & Shoba, 2011 ), glycosides, flavones, and flavonoids (Santhi & Sengottuvel, 2016 ; Siddiqui et al., 2009 ), catecholic and gallic tannins (Njoku & Obi, 2009 ), phenols (Evans, 2009 ), phytosterol (Edeoga et al., 2005 ), saponins (Roghini & Vijayalakshmi, 2018 ), as well as steroids and terpenoids (Awala & Oyetayo, 2015 ). 2.6.2 Quantitative phytochemical assessment 2.6.2.1 Assessment of total flavonoid content The total flavonoids content in the solvent extract of D. metel leaves was assessed using the aluminum chloride colorimetric assay. Briefly, 2 ml of each solvent extract was mixed well with 6ml of methanol and 10ml of purified water. Subsequently, 0.4ml of 10% aluminum chloride (AlCl₃) and 0.2 ml of 1M potassium acetate (CH₃COOK) were added. The solution was left to sit for 30 min in the darkness at room temperature in order to measure the absorbance at 420nm compared to a reagent blank (made without the extract). Distilled water was used to prepare the blank. Next, the completed mixture was moved to a 96-well ELISA microplate, and the absorbance was analyzed at 420 nm with a microplate reader. Quercetin (1 mg/ml) was used to create a calibration curve. Serial dilutions of quercetin (1, 0.50, 0.25, 0.10, 0.05, 0.02, 0.01, and 0 mg/ml) were prepared to develop the standard curve. From this curve, the total flavonoid content was calculated and expressed as quercetin equivalents (QE) in mg/g of the extract (Aiyegoro & Okoh, 2010 ; Dehpour et al., 2009 ). All treatments were replicated ten times. 2.6.2.2 Assessment of total phenolic content The Folin-Ciocalteu reagent technique was used to evaluate the overall phenolic levels in the solvent extract of D. metel leaves. In summary, approximately 2mL of every solvent extract was combined with 4mL of 2% Sodium carbonate (Na2CO3) and poured into 5mL of 10% Folin-Ciocalteu’s reagent (C 10 H 5 NaO 5 S). The solution was mixed and left to incubate at room temperature for 15 min in the dark. Afterwards, the mixture solution was then added into a 96-well ELISA microplate for measuring absorbance at 765nm using plate readers (SpectraMax-190, made in China, developed in the USA). A standard 1 mgml − 1 gallic acid solution was utilized to create a calibration curve, using concentrations of 1, 0.50, 0.25, 0.10, 0.05, 0.02, 0.01, and 0 mgml − 1 . The phenolic content in the extract was determined from the calibration curve and expressed as Gallic acid equivalents (GAE) in mgg − 1 (Aiyegoro & Okoh, 2010 ). Each of the treatments was replicated ten times. 2.8 Statistical Analysis ANOVA was employed to examine the data, and the significance of the mean difference between treatments was evaluated with the Duncan's Multiple Range Test (DMRT) at a significance level of P < 0.05. The examination was carried out using IBM-SPSS software, particularly version 25.0. 3. Results 3.1. Extraction of Yield The powdered samples were extracted using a cold extraction method with ethanol (EtOH), ethyl acetate (EA), dichloromethane (DCM), n-hexane (n-Hx), and distilled water (DW) as shown in Figure-3.1. The results indicated that the highest extract yield was obtained with distilled water at 14.57%, followed by ethanol at 9.20%. In contrast, n-hexane produced the lowest yield at 1.62% as shown in Table-3.1. Table-3.1: Yield of Leaves’ extracts of Datura metel Extract Yield (%) EtOH 9.20 ± 0.16 b EA 4.44 ± 0.0 c DCM 2.47 ± 0.09 d n-Hx 1.62 ± 0.6 e D.W 14.57 ± 0.43 a Statics S.S = 583.75 M.S = 145.94 Df = 4 f = 3186*** Data in the columns is described as mean values ± standard deviations with various superscripts are significantly different according to DMRT” P > 0.05). S.S (Sum of square); M.S (Mean square); Df (Degree of freedom); f (Significance); *** (significance level). 3.2. Biochemical Analysis 3.2.1 GC-MS Analysis of chemical compounds Gas Chromatography-Mass Spectrometry (GC-MS) analysis was conducted using the Agilent 6890-5973N system. The analysis identified ten chemical compounds, with 11H-Pyrido [3',2':4,5] imidazo [2,1-b] [1,3] benzothiazin-11-one exhibiting the highest peak area at 29.76%. This was followed by 1,3-dimethylbenzene at 18.35%, 1,3,5-trimethylbenzene at 11.73%, and didodecyl phthalate at 10.90% as major compounds. Additionally, six compounds were classified as minor, with peak areas ranging from 3.87–6.91% as illustrated in Table-3.2. Table-3.2: Chemical composition of leaves extract of Datura metel Pk. # RT Area% Name of Compounds M. F M. Wt. 1 3.23 18.35 Benzene, 1,3-dimethyl- C 8 H 10 106.17 2 3.93 10.09 1,3,5-Trimethylbenzene C 6 H 3 (CH 3 ) 3 120.19 3 4.01 3.87 Acetaldehyde C 2 H 4 O 44.05 4 4.08 4.01 Cyclotetrasiloxane, octamethyl- C 8 H 24 O 4 Si 4 296.61 5 4.43 4.46 Neophytadiene C 20 H 38 278.5 6 5.52 6.91 Undecane C 11 H 24 156.31 7 6.88 4.08 L-Alanine-4-nitroanilide C 9 H 11 N 3 O 3 209.2 8 14.29 29.76 11H-Pyrido[3',2':4,5]imidazo [2,1-b][1,3]benzothiazin-11-one C 13 H 7 N 3 OS 253.28 9 15.84 6.72 4-Fluorohistamine C 5 H 8 FN 3 129.14 10 17.59 11.73 Didodecyl phthalate C 32 H 54 O 4 502.8 M.F (Molecular Formula; M.W (Molecular Weight); R.T (Retention Time) 3.3 Assessment of insecticidal activity The mortality of Brevicoryne brassicae treated with various concentrations of Datura metel extract through the leaf dip method significantly increased with both concentration and exposure time. At 12 h post-treatment, the mortality rates ranged from 6.66% at 12.5 mg/mL to 33.33% at 200 mg/mL, with the highest mortality of 78.33% observed for the positive control (imidacloprid). The control group (CK) showed negligible mortality (1.66%). By 24 h, mortality rates exhibited a substantial increase. The lowest concentration (12.5 mg/mL) resulted in 15.00% mortality, whereas the highest concentration (200 mg/mL) achieved 73.33%, approaching the efficacy of the positive control at 85.00%. At 48 h, the trend persisted, with the mortality rates further increasing across all concentrations. The 200 mg/mL concentration exhibited 73.33% mortality, while the positive control recorded the highest mortality at 95.00%. The untreated control group (CK) remained statistically insignificant, with 1.66% mortality. Statistical analysis revealed a highly significant (p < 0.05) difference in mortality rates across all treatments, as indicated by the F-values (145.778, 151.417, and 201.610 for 12, 24, and 48 hours, respectively) as shown in Figure-3.2. The mean mortality of Brevicoryne brassicae treated with Datura metel extract at different concentrations (12.5, 25, 50, 100, and 200 mg/mL) and evaluated at 12, 24, and 48 h post-treatment using the aphid dip method is presented in Fig. 2. A significant dose- and time-dependent increase in mortality was observed across all concentrations (p ≤ 0.05). At 12 h post-treatment, mortality ranged from 18.33% at the lowest concentration (12.5 mg/mL) to 43.33% at the highest concentration (200 mg/mL). Mortality in the positive control (Imida) was 83.33%, significantly higher than all other treatments. The untreated control (CK) showed no mortality. At 24 h post-treatment, mortality increased across all concentrations, with 26.66% mortality at 12.5 mg/mL and 58.33% at 200 mg/mL. The positive control resulted in 91.66% mortality, whereas the untreated control showed negligible mortality (1.66%). At 48 h post-treatment, the trend persisted with the highest mortality observed at 200 mg/mL (81.66%) among the Thorn apple treatments, while the positive control (Imida) showed 98.33% mortality. The untreated control showed minimal mortality (5.00%). Statistical analysis (DMRT) indicated significant differences among treatments at all-time points (F = 213.708, 281.238, and 281.458; p = 0.000). These results highlight the effectiveness of Thorn apple extract in controlling B. brassicae , particularly at higher concentrations, though the mortality remained lower than the positive control (Imida). The untreated control consistently exhibited the lowest mortality as shown in Figure-3.3. 3.4 Phytochemical analysis 3.4.1 Quantitative evaluation of Phytochemicals The contents of total phenols, flavonoids, and DPPH scavenging activity demonstrated by the leaf extract of D. metel are presented in Table-3.5. The findings revealed that the highest total phenol content was found in the D.W extract, at 75.03 GAE/g, while the lowest content was in the DCM extract at 2.60 GAE/g. In contrast, the ethyl acetate extract exhibited the highest total flavonoid content at 99.32 mg quercetin equivalent (QE) per gram, followed by the DCM extract at 69.86 mg QE/g. Additionally, the inhibition percentages indicated that all extracts effectively scavenged free radicals, demonstrating significant antioxidant activity with values of 81.59%, 75.31%, 66.61%, 64.06%, and 63.9% for D.W, n-Hx, EtOH, EA, and DCM, respectively. Table-3.5: Total Phenols, Flavonoid contents and DPPH inhibition % of Datura metel extract Extracts Total phenolic contents mg GAE/g Total flavonoid contents mg QE/g DPPH Inhibition (%) EtOH 17.66 ± 0.09 b 18.56 ± 0.11 c 66.61 ± 0.12 c EA 16.476 ± 0.64 c 99.32 ± 0.86 a 64.06 ± 0.59 d DCM 2.60 ± 0.02 e 69.86 ± 0.01 b 63.91 ± 0,.07 d n-Hx 8.19 ± 0.18 d 12.86 ± 0.07 d 75.31 ± 0.13 b D.W 75.03 ± 0.15 a 5.45 ± 0.06 e 81.59 ± 0.96 a Statics S.S = 17489 S.S = 310178 S.S = 1230 M.S = 4372 M.S = 77544 M.S = 307 Df = 4 Df = 4 M.S = 4 f = 46192*** f = 504815*** f = 1174*** Data in the columns is described as mean values ± standard deviations with various superscripts are significantly different according to DMRT” P > 0.05). S.S (Sum of square); M.S (Mean square); Df (Degree of freedom); f (Significance); *** (significance level). 3.4.2 Qualitative evaluation of phytochemicals The qualitative analysis of the phytochemicals revealed the presence of various phytoconstituents in the extract, including alkaloids, glycosides, flavones, flavonoids, terpenoids, steroids, phenols, phytosterol, saponins, terpenoids, quinones and gallic tannins. However, anthraquinones and betacyanin were found to be present in negligible amounts. The results for the phytochemicals screening are presented in Table-3.6. Table-3.6: Qualitative phytochemicals screening of Datura metel leave’s extract Phytoconstituents EtOH EA DCM n-Hx D.W Alkaloids +++ ++ ++ - - Antraquinones - - - + - Betacyanin + - - - - Catecholic tannins +++ - - - + Coumarins + - + - + Flavones ++ +++ +++ ++ + Flavonoids ++ +++ +++ ++ + Gallic tannins +++ +++ - - - Glycosides - + ++ + + Phenols ++ ++ + + +++ Phytosterol ++ ++ ++ + + Saponins + ++ - - ++ Steroids + + + + ++ Terpenoids - ++ ++ + +++ Quinones + + - - + Highly present (+++); moderately present (++); slightly present (+); absence (-) 4. Discussions Plants are enriched with naturally occurring compounds known as phytochemicals, which serve as powerful protectors for the plants themselves. Similarly, these compounds, often referred to as secondary metabolites, can also benefit humans by offering potential for disease prevention and treatment. Regular consumption of a diet rich in these biochemical compounds can enhance our health and strengthen our defences against heart disease and other chronic illnesses (Vasu et al., 2009 ). Therefore, this study was designed to assess the chemical profiling, antioxidant activities, and phytochemical analysis of leaf extracts from Datura metel . GC-MS analysis revealed the presence of bioactive phytoconstituents, with peak area percentages, retention times, molecular formulas, and molecular weights of the identified compounds being assessed. The major bioactive compounds identified included 11H-Pyrido [3',2':4,5] imidazo [2,1-b] [1,3] benzothiazin-11-one, which exhibited the highest peak area at 29.78%, followed by Benzene, 1,3-dimethyl- at 18.35%, 1,3,5-Trimethylbenzene at 11.73%, and Didodecyl phthalate at 10.9%. These compounds exhibit various biological activities; for instance, neophytadiene is known for its antimicrobial, antioxidant, antipyretic, and, anti-inflammatory properties, and has been utilized in the treatment of skin diseases, rheumatism, and headaches (Riaz et al., 2023 ). Hexadecanoic acid, recognized as an anti-inflammatory fatty acid, also exhibits antifungal and antibacterial activity (Hrichi et al., 2022 ). Furthermore, Hexadecanoic acid, 1-pentadecene, and 1-tetradecanol demonstrate antimicrobial activity against Aspergillus terreus , Aspergillus niger , and Aspergillus flavus (Behiry et al., 2022 ). 1,3-dimethyl benzene, also known as m-xylene, can be used as an inert component in non-food pesticide items as per the Environmental Protection Agency (EPA) Chemical and Products Database (CPDat). Mesitylene, also known as 1,3,5-Trimethylbenzene, is commonly used in labs as a unique solvent and frequently used as a reference standard in NMR spectroscopy. Undecane serves as a sex pheromone for certain insects and ants, including Formica lugubris and Caloglyphus rodriguezi (Walter et al., 1993 ), and is also a component of the attraction pheromone of Oecophylla longinoda . Additionally, acetaldehyde, a member of the aldehyde group, encompasses a range of highly reactive and hazardous chemicals with diverse industrial applications, including cosmetics, pharmaceuticals, agrochemicals, plastic manufacturing, biofuels, and fragrance production (Aljaafari et al., 2022 ). The GC-MS analysis identified several bioactive phytoconstituents with significant biological activities, highlighting their potential applications in various fields, including medicine and industry. Regarding extraction efficiency, distilled water exhibited the highest extraction efficiency (14.57%), followed by ethanol (9.20%) and ethyl acetate (4.44%), which is consistent with the findings of (Prasathkumar et al., 2022 ). Phytochemical analysis of D. metel demonstrates the presence of various compounds, including flavonoids, saponins, alkaloids, and phenolic substances such as tannins, aligning with the results reported by (Arowora et al., 2016 ). Because plants co-evolved with the insects that eat them, plants have evolved defense mechanisms. These mechanisms involve the synthesis of substances that disrupt the typical physiological and behavioral features of insects. Consequently, these substances affect feeding, mating, mortality, and egg laying, among other facets of insect life (Adesina, 2022 ). Natural plant-based essential oils and chemical compounds have a variety of qualities, such as cytotoxicity, antioxidant activity, antifungal and insecticidal, and antibacterial functionality (Sharma et al., 2023 ). The LD 50 value of 2.962 mlKg -1 from D. stramonium seed extracts was shown by (Jawalkar et al., 2016 ), indicating the potential of seed extract against the rice weevil, S. oryzae. Furthermore, it was observed that D. metel extract was highly toxic to Fall army worms (FAW) at varying concentrations, such as 60% and 100% in distilled water (Maurawa et al., 2021 ). It is evident that Datura emits a particular scent that has insect-repelling properties.(Acheuk & Doumandji-Mitiche, 2013 ) reported similar results, stating that alkaloids disrupt neuroendocrine regulation by deactivating acetylcholinesterase in treated larvae. However, using natural products on crops could be risky if one knows a little bit about their toxicity (Bateman et al., 2018 ). It is clear that Datura emits a particular scent that has insect-repelling properties. According to (Gaire & Subedi, 2013 ), the presence of phytochemicals like alkaloids in D. strumonium extract results in a distinctive smell that serves as a deterrent for a variety of insect pests. (Acheuk & Doumandji-Mitiche, 2013 ) reported similar results, stating that alkaloids disrupt neuroendocrine regulation by deactivating acetylcholinesterase in treated larvae. However, using natural products on crops or as personal protective measures could be risky if one knows a little bit about their toxicity (Bateman et al., 2018 ). According to some earlier research, when D. stramonium root extract was used at 100% concentration, it showed a distinct larvicidal efficacy ranging from 50–100% within 24 hours of exposure (Singh et al., 2023 ). (Sharma et al., 2023 ), reported the potential of extracts of the roots, leaves, flowers, and seeds of Datura alba and Datura stramonium have significant insecticidal properties. Numerous studies indicate that plants possess antioxidant attributes primarily due to their phenolic content (Nwozo et al., 2023 ). Antioxidants counteract free radicals, prevent oxidative damage, sequester metal ions, and act as oxygen scavengers. Among the solvent extracts, the distilled water extract contained the highest levels of phytochemicals, with total phenolic content measuring 75.03 GAEg⁻¹, followed by ethanol at 17.66 GAEg⁻¹ and ethyl acetate at 16.47 GAE g⁻¹. However, the maximum flavonoid content was observed in the ethyl acetate extract (99.32 mgQEg -1 ), followed by dichloromethane (69.86 mgQEg -1 ). Various research studies have shown that alkaloids, phenolic compounds, and flavonoids derived from plants contribute to antioxidant effects (Alam et al., 2021 ), while, phenolic compounds are known for their capacity to inhibit oxidative changes by counteracting free radicals, absorbing oxygen, or decomposing peroxides (Nijveldt et al., 2001). Distilled water proved to be the most effective solvent for extracting phytochemicals from D. metel , yielding the highest extraction efficiency and total phenolic content, while also demonstrating significant antioxidant properties. Consequently, the evaluation of phytochemicals is important in phytomedicine, aiding in drug research and advancement (Chihomvu et al., 2024 ). It is important to mention that flavonoids, a type of secondary metabolites, have antibacterial, antioxidant, and anti-aging characteristics and could be useful in cancer treatment by preventing carcinogenesis at different phases. According to Mueed et al., ( 2023 ), Tannins, which are polyphenols soluble in water, show antimicrobial characteristics. Secondary compounds from plants, such as tannins, phenols, flavonoids, saponins, and essential oils have been shown to possess antioxidant and antimicrobial capabilities, providing defence against harmful infections (Tungmunnithum et al., 2018 ). The results showed that each extract exhibited significant DPPH inhibition, with distilled water extract leading with 81.59% inhibition, followed by n-hexane (75.30%), ethanol (66.61%), ethyl acetate (64.06%), and dichloromethane (63.91%). Evaluating phytochemicals, such as flavonoids and tannins, is important for their use in drug development, disease prevention, and treatment due to their strong antioxidant and antimicrobial properties. Herbal plants containing phenolic compounds and natural antioxidants are employed to counteract oxidative stress by scavenging free radicals (Akbari et al., 2022 ). Natural antioxidants are crucial for maintaining good health and lowering the risks associated with chronic diseases by combatting oxidative damage caused by reactive oxygen species (ROS) or environmental oxygen (Sharifi-Rad et al., 2020 ). Fruits, vegetables, leaves, and whole grains are common providers of antioxidants like carotenes, phenolic acids, phytoestrogens, vitamin C, and vitamin E, which may lower the risk of disease development (Rahaman et al., 2023 ). Our results support the research conducted by Prasathkumar et al., ( 2022 ), demonstrating the notable antioxidant properties of methanolic extracts from D. metel leaves, displaying varying degrees of radical scavenging activity between 48.16% and 93.55% at different concentrations, and an IC 50 value of 146.53 µgmL -1 . Herbal plants, specifically D. metel , exhibit distinguished antioxidant properties by containing abundant natural antioxidants and phenolic compounds, which can effectively combat oxidative stress and possibly lower the chances of developing diseases. Consequently, D. metel exhibits promising antioxidant, antimicrobial, and insecticidal activities, highlighting its potential as a source of bioactive phytochemicals. However, further research is required to isolate and identify specific active compounds, and in vivo studies to deepen our understanding of their mechanisms of action as antioxidants and their various biological activities. Conclusion The present study findings suggest that the leaves of Datura metel contain a substantial range of chemical compounds, including valuable phytoconstituents such as phenols and flavonoids. Furthermore, GC-MS analysis revealed the presence of significant amounts of these compounds, which are associated with free radical scavenging activity. Similarly, the extracts displayed significant insecticidal activity against B. brassicae . However, further studies are necessary to isolate and identify these biochemical compounds for potential applications in the pharmaceutical and agrochemical sectors, particularly as alternative medicines and pesticides, respectively. Declarations Contribution of Authors: Conceptualization, M.J.; Formal analysis, M.A.; A.R. and S.A. Investigation, M.J.; Methodology, M.A.; K.F.A.; A.J.; Project administration, M.J.; Software, M.A.; A.M., and S.F.; Supervision, M.J.; A.E.S.; Visualization, M.A.; A.M., and M.J.; Writing – original draft, M.A.; Writing – review & editing, S.R. A.A.; and S.A. Funding: National Key Research & Development Program of China (2016YFD0200500) granted research funding for this project. The authors extend their appreciation to Researchers Supporting Project number (RSPD2025R561), King Saud University, Riyadh, Saudi Arabia. Conflicts of Interest: There is no conflict of interest by any author. Acknowledgments: Professor Ji Mingshan provided support & supervision and the experimental guidelines of lab mates of Biopesticides Laboratory, Plant Protection College is greatly acknowledged. The authors extend their appreciation to Researchers Supporting Project number (RSPD2025R561), King Saud University, Riyadh, Saudi Arabia. References Acheuk, F., & Doumandji-Mitiche, B. (2013). 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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-6110776","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":423516221,"identity":"9cfd720a-1f75-4669-aa5f-8df3480df28a","order_by":0,"name":"Abdul Mateen","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABDklEQVRIie2PMWsCMRTHX3iQLqmdD9t+g8IVQRGln8Vw0K5ONwuFdDlxtdN9BUfHSNZ4WQu3KH4BS5cWEfqudYwWnYrkt7yXP/nlvQAEAv8RDYDA4Xp37AKIqnLYNfsVURXikW7icYr5W6mVmn30UyPuXtxs8T51Ms+chnVqoCW0V4mKHtbHhRFNm+D9qy3lZP4MjBJoDwdeJbYA9UtFik44NaRcISAlEDv/YqTg5kdxq4vNVhUyH5GyPazw3ylvCUemtBzQYtSQMvcvFlmmOuPiiZRVIxqqpDGxJp5llLQz//drFk3ZTzu3TSeX6y/1cJNncrn4pKQlev7NgHnGV8+LeI9wgBOUQCAQOE++AfT9YHOHpXMDAAAAAElFTkSuQmCC","orcid":"","institution":"University of Sialkot","correspondingAuthor":true,"prefix":"","firstName":"Abdul","middleName":"","lastName":"Mateen","suffix":""},{"id":423516222,"identity":"d52a526d-f823-43fa-8e53-68dbcb6bd963","order_by":1,"name":"Ansar Javeed","email":"","orcid":"","institution":"Zhejiang Sci-Tech University","correspondingAuthor":false,"prefix":"","firstName":"Ansar","middleName":"","lastName":"Javeed","suffix":""},{"id":423516223,"identity":"e82dda7f-4aa2-4795-843a-bb5adbcf5034","order_by":2,"name":"Adnan Arshad","email":"","orcid":"","institution":"Lanzhou University","correspondingAuthor":false,"prefix":"","firstName":"Adnan","middleName":"","lastName":"Arshad","suffix":""},{"id":423516224,"identity":"697fcd01-d737-41f2-90c4-6b13c4b416b0","order_by":3,"name":"Amer Rasul","email":"","orcid":"","institution":"Directorate General of Pest Warning","correspondingAuthor":false,"prefix":"","firstName":"Amer","middleName":"","lastName":"Rasul","suffix":""},{"id":423516225,"identity":"4074b9c6-ca3f-48a9-b746-0678f313b979","order_by":4,"name":"Shams Ur Rehman","email":"","orcid":"","institution":"Peking University Institute of Advanced Agricultural Sciences","correspondingAuthor":false,"prefix":"","firstName":"Shams","middleName":"Ur","lastName":"Rehman","suffix":""},{"id":423516226,"identity":"bfc5a3df-9569-4bbc-96d5-ab32760b95fe","order_by":5,"name":"Sabin Fatima","email":"","orcid":"","institution":"University of Agriculture Faisalabad","correspondingAuthor":false,"prefix":"","firstName":"Sabin","middleName":"","lastName":"Fatima","suffix":""},{"id":423516227,"identity":"e0333804-9ac8-4517-af63-f246032a6990","order_by":6,"name":"Shiza Ali","email":"","orcid":"","institution":"University of Agriculture Faisalabad","correspondingAuthor":false,"prefix":"","firstName":"Shiza","middleName":"","lastName":"Ali","suffix":""},{"id":423516228,"identity":"ffa8658b-3a52-43da-8859-c6a3e8d204a1","order_by":7,"name":"Mingshan Ji","email":"","orcid":"","institution":"Shenyang Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Mingshan","middleName":"","lastName":"Ji","suffix":""},{"id":423516229,"identity":"4996ec94-adda-4ad1-93aa-69450d862bdf","order_by":8,"name":"Khalid F. 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(c) Coarsely ground plant material and make a fine powder for bioactive compound extraction. (d, e) Extracts of \u003cem\u003eDatura metel\u003c/em\u003e, utilized for bioactivity testing against \u003cem\u003eBrevicoryne brassicae\u003c/em\u003e.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6110776/v1/f2f04a4ea559ff8dd1964054.png"},{"id":77708442,"identity":"ca7e0586-fe8c-43ca-bb70-85fd386122de","added_by":"auto","created_at":"2025-03-04 12:30:36","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":15733,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure-3.2\u003c/strong\u003e: Mean mortality of \u003cem\u003eBrevicoryne brassicae\u003c/em\u003e caused by \u003cem\u003eDatura metel\u003c/em\u003eextract using the leaf dip method. Data represent the average percentage mortality observed across different extract concentrations. Error bars indicate standard error (SE) of the mean. Different letters indicate significant differences (p \u0026lt; 0.05) among treatments.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6110776/v1/07a25f3d864652e730bc00eb.png"},{"id":77708441,"identity":"79ad2153-6294-4fa4-a939-1f609b3b1b24","added_by":"auto","created_at":"2025-03-04 12:30:35","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":14710,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure-3.3\u003c/strong\u003e: Mean mortality of \u003cem\u003eBrevicoryne brassicae\u003c/em\u003e caused by \u003cem\u003eDatura metel\u003c/em\u003eextract using the aphid dip method. Data represent the average percentage mortality observed across different extract concentrations. Error bars indicate standard error (SE) of the mean. Different letters indicate significant differences (p \u0026lt; 0.05) among treatments.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6110776/v1/3e781e6a136ac6601bdd532e.png"},{"id":77709265,"identity":"51ca9a21-8493-451c-98f3-6c0413fe6fcb","added_by":"auto","created_at":"2025-03-04 12:38:33","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2124569,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6110776/v1/df7bf38a-434d-4f91-b061-70e4d059838d.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Chemical, Insecticidal, Antioxidant and Phytochemical Assessment of Thorn Apple (Datura metel L.) leaf extract from Pakistan's Subtropical Climate","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003ePlants play a vital role in the industrial agrochemical and pharmaceutical sectors, as they contain a wide range of phytochemicals that are effective in treating various diseases. They are the primary sources used in alternative medicine for combating a broad spectrum of ailments. Many traditional medicines come from plants, and around 80% of people worldwide use herbal remedies for maintaining good health. Various plant components such as leaves, flowers, roots, stems, bark, rhizomes, bulbs, and seeds are frequently utilized in making herbal goods (Matotoka \u0026amp; Masoko, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Pliego et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, the use of medicinal plants varies across regions, influenced by factors such as environmental conditions, cultural practices, historical context, individual beliefs, and philosophical perspectives (Ahmad et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Medicinal plants are indispensable resources for both traditional and modern medicine, with their usage made by regional, cultural, and environmental factors.\u003c/p\u003e \u003cp\u003e \u003cem\u003eDatura metel\u003c/em\u003e commonly known as Thorn Apple, is famous for its medicinal properties and its potential use in the agrochemical industry. It is widely cultivated across Asia, Africa, England, India, and various tropical and subtropical regions (Gaire \u0026amp; Subedi, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). However, in Asia, it typically thrives in wild habitats (Bhattacharjee et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). The medicinal properties of \u003cem\u003eDatura metel\u003c/em\u003e are attributed to its bioactive compounds, which have proven effective in treating various conditions, including asthma and bronchitis. Although the plant has narcotic effects, it also offers specific health benefits that make it a valuable a medicinal resource. These benefits include antidiabetic, antimicrobial, anti-inflammatory, anti-asthmatic, antioxidant, insecticidal, cytotoxic, pain-relieving, neurological and, wound healing effects (Alam et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Shah et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). The plant\u0026rsquo;s various parts, especially the leaves, contain bioactive compounds, prompting investigations into its chemical composition, antioxidant and phytoconstituents (Rahaman et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). \u003cem\u003eDatura metel\u003c/em\u003e's diverse medicinal properties and bioactive compounds make it a valuable resource for both the pharmaceutical and agrochemical industries.\u003c/p\u003e \u003cp\u003eIn the agrochemical industry, Datura is recognized for its larvicidal ability to eliminate larvae of the red flour beetle (\u003cem\u003eTribolium castaneum\u003c/em\u003e), its effectiveness as a mosquito repellent, and its use in treating animal bites, including snake bites, by providing pain relief (Devi et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Meselhy, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). It contains substantial amounts of carbohydrates, moisture, fats, ash, proteins, and crude fiber. Additionally, it is rich in important phytochemicals, including phenolic compounds, alkaloids, flavonoids, cardiac glycosides and, tannins (Bagewadi et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Fatima et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Moreover, the significant role of secondary metabolites from the plant kingdom in various biological processes is well established (Divekar et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Therefore, identifying these valuable phytoconstituents is crucial for understanding their bioactive properties (Xiao \u0026amp; Bai, \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Thus, rich phytochemical profile and bioactive properties of this plant make it a valuable resource for both agricultural and medicinal applications.\u003c/p\u003e \u003cp\u003ePharmacologically consumption of any part of the Datura plant can cause severe anticholinergic effects, potentially leading to toxicity. While the entire plant contains toxic properties, the seeds are especially harmful. Notably, neither drying nor boiling the plant eliminates its toxic characteristics (Setshogo, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Despite its toxicity, \u003cem\u003eD. metel\u003c/em\u003e also possesses therapeutic applications, making it a subject of extensive research (Naik et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Xu et al., \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThis study highlights \u003cem\u003eD. metel\u003c/em\u003e as a promising source of bioactive compounds with significant biological properties. By evaluating the chemical composition and antioxidant activity across various extraction methods, valuable insights into the plant's therapeutic potential were gained. The identification of key phytochemicals and their concentrations enhances our understanding of \u003cem\u003eD. metel\u003c/em\u003e's medicinal applications. Given its traditional use in herbal medicine, these findings may guide future research into its role in developing natural remedies and therapeutic agents, underscoring its importance in pharmacological research for disease prevention and treatment.\u003c/p\u003e"},{"header":"2. Material and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Collection and authentication of samples\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eDuring June-July 2024, \u003cem\u003eD. metel\u003c/em\u003e leaves were collected from Punjab, Pakistan at the coordinates 32.66843\u0026deg;N and 74.17298\u0026deg;E. Specimens were collected from the identical mature, fully blossoming plant, assuring that only healthy leaves were chosen. This area has a humid subtropical climate in accordance with the Koppen climate classification, which includes four separate seasons. Temperatures in this region fluctuate between 42\u0026deg;F and 103\u0026deg;F annually. The plant species was scientifically verified by Dr. Dilbar Hussain at Ayub Agriculture Research Institute (AARI) Entomological Research Institute Faisalabad Pakistan.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Extraction procedure\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eThe samples were left at room temperature to avoid photo-oxidation until fully dried and were subsequently pulverized into a fine powder with an electric blender. Various solvents such as ethanol (EtOH), ethyl acetate (EA), dichloromethane (DCM), n-hexane (n-Hx), and distilled water (DW) were used to extract the dried powder for 72 hours using a cold extraction method at room temperature. In the extraction procedure, 10g of every dried sample was separately extracted with 100 mL of each solvent, keeping a ratio of 1:10 between sample and solvent (Li Ping et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) stated. The extraction procedure was repeated on three times. The contents extracted were sieved through Whatman filter paper No. 1 and condensed with a rotary evaporator (model R-210 BUCHI Labortechnik AG, CH-9230 Flawil 1/Switzerland) to decrease the volume. After allowing the solvents to completely evaporate in a fume hood for 24 hours, the quantity produced was determined using the Equation (Eq.\u0026nbsp;1). The samples collected were kept in glass-stoppered vials at 4\u0026deg;C for future phytochemical analysis and evaluation of antioxidant potential (Mueed et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cspan class=\"InlineEquation\"\u003e \u003cspan class=\"mathinline\"\u003e\\(\\:\\varvec{Y}\\varvec{i}\\varvec{e}\\varvec{l}\\varvec{d}\\:\\left(\\varvec{\\%}\\right)=\\frac{\\:\\mathbf{W}\\mathbf{e}\\mathbf{i}\\mathbf{g}\\mathbf{h}\\mathbf{t}\\:\\mathbf{o}\\mathbf{f}\\:\\mathbf{t}\\mathbf{h}\\mathbf{e}\\:\\mathbf{e}\\mathbf{x}\\mathbf{t}\\mathbf{r}\\mathbf{a}\\mathbf{c}\\mathbf{t}\\:\\left(\\mathbf{g}\\right)}{\\:\\mathbf{W}\\mathbf{e}\\mathbf{i}\\mathbf{g}\\mathbf{h}\\mathbf{t}\\:\\mathbf{o}\\mathbf{f}\\:\\mathbf{t}\\mathbf{h}\\mathbf{e}\\:\\mathbf{d}\\mathbf{r}\\mathbf{i}\\mathbf{e}\\mathbf{d}\\:\\mathbf{s}\\mathbf{a}\\mathbf{m}\\mathbf{p}\\mathbf{l}\\mathbf{e}\\:\\left(\\mathbf{g}\\right)\\:}\\times\\:100\\varvec{\\%}\\)\u003c/span\u003e \u003c/span\u003e (Eq.\u0026nbsp;1)\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Analysis of chemical compounds\u003c/h2\u003e \u003cp\u003eGC-MS was performed on the solvent extract of \u003cem\u003eD. metel\u003c/em\u003e leaves using an Agilent 6890-5973N system to analyze the chemical compounds. The gas chromatograph was set up using an HP1 capillary column (Model Number: TG-5MS; 30 m \u0026times; 250 \u0026micro;m \u0026times; 0.25 \u0026micro;m) and run with a flow rate of 1.0 mL/min. The oven started at 70\u0026deg;C for 0 min, increasing at 3\u0026deg;C/min until reaching 220\u0026deg;C in 10 min. The temperature at the inlet is 250\u0026deg;C, divided at a ratio of 10:1. Included in the additional thermal parameters were a thermal auxiliary temperature of 285\u0026deg;C and a mass spectrometry (MS) scan range from 35 to 520 automatic mass unit (AMU). The MS source was maintained at a temperature of 230\u0026deg;C, with the quadruple temperature held constant at 150\u0026deg;C. Helium served as the carrier gas, flowing at a rate of 1.0mL/min, while HPLC-grade methanol was utilized as the reference sample. The components in the sample were confirmed and authenticated by cross-referencing them with information from the National Institute of Standards and Technology (NIST) Mass Spectral Database (Sparkman, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2005\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Insecticidal activity\u003c/h2\u003e \u003cp\u003eInsecticidal activity of the \u003cem\u003eD. metel\u003c/em\u003e extract was evaluated using two different bioassays like standard leaf dip and individual dip and methods.\u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section3\"\u003e \u003ch2\u003e2.4.1 Leaf dip assay\u003c/h2\u003e \u003cp\u003eUsing acetone as solvent, already prepared extracts were serially concentrated in Tween-20 (1%) solution at 12.5, 25, 50, 100, and 500 \u0026micro;gmL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. Adult wingless aphids were collected from maize plants that had not been treated with insecticides. In short, fresh maize leaf discs were measuring 5 cm in diameter were cut off, dipped in the corresponding concentration for 10 seconds, and then allowed to dry at room temperature for 5 minutes. A fine-haired aphid was used to carefully release 10 adults, healthy aphids onto the leaf disc in the Petri dish and then incubated for 48 h at 65% relative humidity (R.H), 25˚C temperature, and with a 16:8 (light: dark) photoperiod.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e2.4.2 Aphid dip assay\u003c/h2\u003e \u003cp\u003eIn a similar manner, for the aphid dip method10 adult, healthy aphids were carefully dipped in the corresponding concentrations for 5 s before being released on extract free cabbage leaves. The same adjustments were made to the incubation period, R.H., temperature, and photoperiod as with the leaf dip method. The positive control, Imidacloprid 25% WP, was used at a rate of 0.0025 mLmL\u0026thinsp;\u0026minus;\u0026thinsp;1 in conjunction with control (CK) using a 1% Tween-20 solution. A binocular microscope was used to collect mortality data at 12, 24, and 48 h, and needle probing was used to observe the aphids' reaction. Five duplicates of each treatment were used.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Assessment of DPPH radical scavenging assay\u003c/h2\u003e \u003cp\u003eThe method proposed by Yu et al., (\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2003\u003c/span\u003e) was followed by applying DPPH radical scavenging assay in a 1% Tween 20 solution to measure antioxidant activity in the solvent extract of \u003cem\u003eD. metel\u003c/em\u003e leaves. Roughly 0.5 mL of the solvent extract was mixed with 3.5 mL of a recently prepared methanol solution containing DPPH (0.004 g/100 mL) in pure methanol of grade HPLC. Following this, 0.25 mL of the extract solution, which had been prepared individually in methanol for each solvent extract, was combined with the freshly made 3.5 mL DPPH solution. The mixture was vigorously shaken to ensure thorough mixing, followed by incubation in darkness at 28\u0026deg;C for 30 min (Roghini \u0026amp; Vijayalakshmi, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Following the incubation period, the combination was moved into a 96-well ELISA microplate for absorbance measurement at 517 nm with a photometric plate reader (Spectra-Max 190, Molecular Devices, Shanghai China). Methanol was used as the control group, whereas a combination of DPPH and methanol was used as the experimental group. Eq.\u0026nbsp;2 was used to calculate the percentage (inhibition) of the DPPH solution by measuring the decrease in absorbance. Higher free radical scavenging activity is reflected by lower absorbance values, as there is an inverse correlation between absorption and radical scavenging activity (Zhao et al., \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). The calculation of the DPPH inhibition percentage was performed with Eq.\u0026nbsp;2.\u003c/p\u003e\u003cp\u003e\u003cimg src=\"data:image/png;base64,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\" height=\"38\" width=\"356\"\u003e\u003c/p\u003e\u003cp\u003eWhere: \u003cem\u003eA\u003c/em\u003e\u003csub\u003e\u003cem\u003eblank\u003c/em\u003e\u003c/sub\u003e = (control absorbance); \u003cem\u003eA\u003c/em\u003e\u003csub\u003e\u003cem\u003eSample\u003c/em\u003e\u003c/sub\u003e = (samples absorbance).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e2.6 Phytochemical analysis\u003c/h2\u003e \u003cdiv id=\"Sec11\" class=\"Section3\"\u003e \u003ch2\u003e2.6.1 Qualitative phytochemical assessment\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eStandard procedures were followed to identify different compounds present in the solvent extract of \u003cem\u003eD. metel\u003c/em\u003e leaves through qualitative analysis of phytochemicals. These comprised alkaloids (Nortjie et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), anthraquinones (Roghini \u0026amp; Vijayalakshmi, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), betacyanins (Ali et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), coumarins (Jayapriya \u0026amp; Shoba, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2011\u003c/span\u003e), glycosides, flavones, and flavonoids (Santhi \u0026amp; Sengottuvel, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Siddiqui et al., \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), catecholic and gallic tannins (Njoku \u0026amp; Obi, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), phenols (Evans, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), phytosterol (Edeoga et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2005\u003c/span\u003e), saponins (Roghini \u0026amp; Vijayalakshmi, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), as well as steroids and terpenoids (Awala \u0026amp; Oyetayo, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e \u003ch2\u003e2.6.2 Quantitative phytochemical assessment\u003c/h2\u003e \u003cdiv id=\"Sec13\" class=\"Section4\"\u003e \u003ch2\u003e2.6.2.1 Assessment of total flavonoid content\u003c/h2\u003e \u003cp\u003eThe total flavonoids content in the solvent extract of \u003cem\u003eD. metel\u003c/em\u003e leaves was assessed using the aluminum chloride colorimetric assay. Briefly, 2 ml of each solvent extract was mixed well with 6ml of methanol and 10ml of purified water. Subsequently, 0.4ml of 10% aluminum chloride (AlCl₃) and 0.2 ml of 1M potassium acetate (CH₃COOK) were added. The solution was left to sit for 30 min in the darkness at room temperature in order to measure the absorbance at 420nm compared to a reagent blank (made without the extract). Distilled water was used to prepare the blank. Next, the completed mixture was moved to a 96-well ELISA microplate, and the absorbance was analyzed at 420 nm with a microplate reader. Quercetin (1 mg/ml) was used to create a calibration curve. Serial dilutions of quercetin (1, 0.50, 0.25, 0.10, 0.05, 0.02, 0.01, and 0 mg/ml) were prepared to develop the standard curve. From this curve, the total flavonoid content was calculated and expressed as quercetin equivalents (QE) in mg/g of the extract (Aiyegoro \u0026amp; Okoh, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Dehpour et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). All treatments were replicated ten times.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section4\"\u003e \u003ch2\u003e2.6.2.2 Assessment of total phenolic content\u003c/h2\u003e \u003cp\u003eThe Folin-Ciocalteu reagent technique was used to evaluate the overall phenolic levels in the solvent extract of \u003cem\u003eD. metel\u003c/em\u003e leaves. In summary, approximately 2mL of every solvent extract was combined with 4mL of 2% Sodium carbonate (Na2CO3) and poured into 5mL of 10% Folin-Ciocalteu\u0026rsquo;s reagent (C\u003csup\u003e10\u003c/sup\u003eH\u003csup\u003e5\u003c/sup\u003eNaO\u003csup\u003e5\u003c/sup\u003eS). The solution was mixed and left to incubate at room temperature for 15 min in the dark. Afterwards, the mixture solution was then added into a 96-well ELISA microplate for measuring absorbance at 765nm using plate readers (SpectraMax-190, made in China, developed in the USA). A standard 1 mgml\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e gallic acid solution was utilized to create a calibration curve, using concentrations of 1, 0.50, 0.25, 0.10, 0.05, 0.02, 0.01, and 0 mgml\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The phenolic content in the extract was determined from the calibration curve and expressed as Gallic acid equivalents (GAE) in mgg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (Aiyegoro \u0026amp; Okoh, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Each of the treatments was replicated ten times.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e2.8 Statistical Analysis\u003c/h2\u003e \u003cp\u003eANOVA was employed to examine the data, and the significance of the mean difference between treatments was evaluated with the Duncan's Multiple Range Test (DMRT) at a significance level of P\u0026thinsp;\u0026lt;\u0026thinsp;0.05. The examination was carried out using IBM-SPSS software, particularly version 25.0.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Extraction of Yield\u003c/h2\u003e \u003cp\u003eThe powdered samples were extracted using a cold extraction method with ethanol (EtOH), ethyl acetate (EA), dichloromethane (DCM), n-hexane (n-Hx), and distilled water (DW) as shown in Figure-3.1. The results indicated that the highest extract yield was obtained with distilled water at 14.57%, followed by ethanol at 9.20%. In contrast, n-hexane produced the lowest yield at 1.62% as shown in Table-3.1.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eTable-3.1: Yield of Leaves\u0026rsquo; extracts of\u003c/b\u003e \u003cb\u003eDatura metel\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabb\" border=\"1\"\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eExtract\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eYield (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEtOH\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDCM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003en-Hx\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD.W\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStatics\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eS.S\u0026thinsp;=\u0026thinsp;583.75\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eM.S\u0026thinsp;=\u0026thinsp;145.94\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDf\u0026thinsp;=\u0026thinsp;4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ef\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3186***\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eData in the columns is described as mean values\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviations with various superscripts are significantly different according to DMRT\u0026rdquo; \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). S.S (Sum of square); M.S (Mean square); Df (Degree of freedom); \u003cem\u003ef\u003c/em\u003e (Significance); *** (significance level).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Biochemical Analysis\u003c/h2\u003e \u003cdiv id=\"Sec19\" class=\"Section3\"\u003e \u003ch2\u003e3.2.1 GC-MS Analysis of chemical compounds\u003c/h2\u003e \u003cp\u003eGas Chromatography-Mass Spectrometry (GC-MS) analysis was conducted using the Agilent 6890-5973N system. The analysis identified ten chemical compounds, with 11H-Pyrido [3',2':4,5] imidazo [2,1-b] [1,3] benzothiazin-11-one exhibiting the highest peak area at 29.76%. This was followed by 1,3-dimethylbenzene at 18.35%, 1,3,5-trimethylbenzene at 11.73%, and didodecyl phthalate at 10.90% as major compounds. Additionally, six compounds were classified as minor, with peak areas ranging from 3.87\u0026ndash;6.91% as illustrated in Table-3.2.\u003c/p\u003e \u003cp\u003e \u003cb\u003eTable-3.2: Chemical composition of leaves extract of\u003c/b\u003e \u003cb\u003eDatura metel\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabc\" border=\"1\"\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePk. #\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eArea%\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eName of Compounds\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eM. F\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eM. Wt.\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e18.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eBenzene, 1,3-dimethyl-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC\u003csub\u003e8\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e106.17\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e10.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1,3,5-Trimethylbenzene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e3\u003c/sub\u003e(CH\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e120.19\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAcetaldehyde\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC\u003csub\u003e2\u003c/sub\u003eH\u003csub\u003e4\u003c/sub\u003eO\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e44.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCyclotetrasiloxane, octamethyl-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC\u003csub\u003e8\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003eSi\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e296.61\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNeophytadiene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC\u003csub\u003e20\u003c/sub\u003eH\u003csub\u003e38\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e278.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eUndecane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC\u003csub\u003e11\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e156.31\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eL-Alanine-4-nitroanilide\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e11\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e209.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e29.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11H-Pyrido[3',2':4,5]imidazo [2,1-b][1,3]benzothiazin-11-one\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC\u003csub\u003e13\u003c/sub\u003eH\u003csub\u003e7\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eOS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e253.28\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4-Fluorohistamine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC\u003csub\u003e5\u003c/sub\u003eH\u003csub\u003e8\u003c/sub\u003eFN\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e129.14\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e17.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e11.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDidodecyl phthalate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC\u003csub\u003e32\u003c/sub\u003eH\u003csub\u003e54\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e502.8\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eM.F (Molecular Formula; M.W (Molecular Weight); R.T (Retention Time)\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003e3.3 Assessment of insecticidal activity\u003c/h2\u003e \u003cp\u003eThe mortality of \u003cem\u003eBrevicoryne brassicae\u003c/em\u003e treated with various concentrations of \u003cem\u003eDatura metel\u003c/em\u003e extract through the leaf dip method significantly increased with both concentration and exposure time. At 12 h post-treatment, the mortality rates ranged from 6.66% at 12.5 mg/mL to 33.33% at 200 mg/mL, with the highest mortality of 78.33% observed for the positive control (imidacloprid). The control group (CK) showed negligible mortality (1.66%). By 24 h, mortality rates exhibited a substantial increase. The lowest concentration (12.5 mg/mL) resulted in 15.00% mortality, whereas the highest concentration (200 mg/mL) achieved 73.33%, approaching the efficacy of the positive control at 85.00%. At 48 h, the trend persisted, with the mortality rates further increasing across all concentrations. The 200 mg/mL concentration exhibited 73.33% mortality, while the positive control recorded the highest mortality at 95.00%. The untreated control group (CK) remained statistically insignificant, with 1.66% mortality. Statistical analysis revealed a highly significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) difference in mortality rates across all treatments, as indicated by the F-values (145.778, 151.417, and 201.610 for 12, 24, and 48 hours, respectively) as shown in Figure-3.2.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eThe mean mortality of \u003cem\u003eBrevicoryne brassicae\u003c/em\u003e treated with \u003cem\u003eDatura metel\u003c/em\u003e extract at different concentrations (12.5, 25, 50, 100, and 200 mg/mL) and evaluated at 12, 24, and 48 h post-treatment using the aphid dip method is presented in Fig.\u0026nbsp;2. A significant dose- and time-dependent increase in mortality was observed across all concentrations (p\u0026thinsp;\u0026le;\u0026thinsp;0.05). At 12 h post-treatment, mortality ranged from 18.33% at the lowest concentration (12.5 mg/mL) to 43.33% at the highest concentration (200 mg/mL). Mortality in the positive control (Imida) was 83.33%, significantly higher than all other treatments. The untreated control (CK) showed no mortality. At 24 h post-treatment, mortality increased across all concentrations, with 26.66% mortality at 12.5 mg/mL and 58.33% at 200 mg/mL. The positive control resulted in 91.66% mortality, whereas the untreated control showed negligible mortality (1.66%). At 48 h post-treatment, the trend persisted with the highest mortality observed at 200 mg/mL (81.66%) among the Thorn apple treatments, while the positive control (Imida) showed 98.33% mortality. The untreated control showed minimal mortality (5.00%). Statistical analysis (DMRT) indicated significant differences among treatments at all-time points (F\u0026thinsp;=\u0026thinsp;213.708, 281.238, and 281.458; p\u0026thinsp;=\u0026thinsp;0.000). These results highlight the effectiveness of Thorn apple extract in controlling \u003cem\u003eB. brassicae\u003c/em\u003e, particularly at higher concentrations, though the mortality remained lower than the positive control (Imida). The untreated control consistently exhibited the lowest mortality as shown in Figure-3.3.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003e3.4 Phytochemical analysis\u003c/h2\u003e \u003cdiv id=\"Sec22\" class=\"Section3\"\u003e \u003ch2\u003e3.4.1 Quantitative evaluation of Phytochemicals\u003c/h2\u003e \u003cp\u003eThe contents of total phenols, flavonoids, and DPPH scavenging activity demonstrated by the leaf extract of \u003cem\u003eD. metel\u003c/em\u003e are presented in Table-3.5. The findings revealed that the highest total phenol content was found in the D.W extract, at 75.03 GAE/g, while the lowest content was in the DCM extract at 2.60 GAE/g. In contrast, the ethyl acetate extract exhibited the highest total flavonoid content at 99.32 mg quercetin equivalent (QE) per gram, followed by the DCM extract at 69.86 mg QE/g. Additionally, the inhibition percentages indicated that all extracts effectively scavenged free radicals, demonstrating significant antioxidant activity with values of 81.59%, 75.31%, 66.61%, 64.06%, and 63.9% for D.W, n-Hx, EtOH, EA, and DCM, respectively.\u003c/p\u003e \u003cp\u003e \u003cb\u003eTable-3.5: Total Phenols, Flavonoid contents and DPPH inhibition % of\u003c/b\u003e \u003cb\u003eDatura metel\u003c/b\u003e \u003cb\u003eextract\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabd\" border=\"1\"\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eExtracts\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTotal phenolic contents mg GAE/g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTotal flavonoid contents mg QE/g\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDPPH\u003c/p\u003e \u003cp\u003eInhibition (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEtOH\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e17.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e66.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e16.476\u0026thinsp;\u0026plusmn;\u0026thinsp;0.64\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e99.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.86\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e64.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.59\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDCM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e69.86\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e63.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0,.07\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003en-Hx\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12.86\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e75.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD.W\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e75.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e81.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.96\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStatics\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eS.S\u0026thinsp;=\u0026thinsp;17489\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS.S\u0026thinsp;=\u0026thinsp;310178\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eS.S\u0026thinsp;=\u0026thinsp;1230\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eM.S\u0026thinsp;=\u0026thinsp;4372\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eM.S\u0026thinsp;=\u0026thinsp;77544\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eM.S\u0026thinsp;=\u0026thinsp;307\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDf\u0026thinsp;=\u0026thinsp;4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eDf\u0026thinsp;=\u0026thinsp;4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eM.S\u0026thinsp;=\u0026thinsp;4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ef\u003c/em\u003e\u0026thinsp;=\u0026thinsp;46192***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003ef\u003c/em\u003e\u0026thinsp;=\u0026thinsp;504815***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003ef\u003c/em\u003e\u0026thinsp;=\u0026thinsp;1174***\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eData in the columns is described as mean values\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviations with various superscripts are significantly different according to DMRT\u0026rdquo; \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). S.S (Sum of square); M.S (Mean square); Df (Degree of freedom); \u003cem\u003ef\u003c/em\u003e (Significance); *** (significance level).\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e \u003ch2\u003e3.4.2 Qualitative evaluation of phytochemicals\u003c/h2\u003e \u003cp\u003eThe qualitative analysis of the phytochemicals revealed the presence of various phytoconstituents in the extract, including alkaloids, glycosides, flavones, flavonoids, terpenoids, steroids, phenols, phytosterol, saponins, terpenoids, quinones and gallic tannins. However, anthraquinones and betacyanin were found to be present in negligible amounts. The results for the phytochemicals screening are presented in Table-3.6.\u003c/p\u003e \u003cp\u003e \u003cb\u003eTable-3.6: Qualitative phytochemicals screening of\u003c/b\u003e \u003cb\u003eDatura metel\u003c/b\u003e \u003cb\u003eleave\u0026rsquo;s extract\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabe\" border=\"1\"\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhytoconstituents\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEtOH\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eEA\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDCM\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003en-Hx\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eD.W\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAlkaloids\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAntraquinones\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBetacyanin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCatecholic tannins\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCoumarins\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFlavones\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFlavonoids\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGallic tannins\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGlycosides\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhenols\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhytosterol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSaponins\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSteroids\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTerpenoids\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eQuinones\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eHighly present (+++); moderately present (++); slightly present (+); absence (-)\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"4. Discussions","content":"\u003cp\u003ePlants are enriched with naturally occurring compounds known as phytochemicals, which serve as powerful protectors for the plants themselves. Similarly, these compounds, often referred to as secondary metabolites, can also benefit humans by offering potential for disease prevention and treatment. Regular consumption of a diet rich in these biochemical compounds can enhance our health and strengthen our defences against heart disease and other chronic illnesses (Vasu et al., \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Therefore, this study was designed to assess the chemical profiling, antioxidant activities, and phytochemical analysis of leaf extracts from \u003cem\u003eDatura metel\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eGC-MS analysis revealed the presence of bioactive phytoconstituents, with peak area percentages, retention times, molecular formulas, and molecular weights of the identified compounds being assessed. The major bioactive compounds identified included 11H-Pyrido [3',2':4,5] imidazo [2,1-b] [1,3] benzothiazin-11-one, which exhibited the highest peak area at 29.78%, followed by Benzene, 1,3-dimethyl- at 18.35%, 1,3,5-Trimethylbenzene at 11.73%, and Didodecyl phthalate at 10.9%. These compounds exhibit various biological activities; for instance, neophytadiene is known for its antimicrobial, antioxidant, antipyretic, and, anti-inflammatory properties, and has been utilized in the treatment of skin diseases, rheumatism, and headaches (Riaz et al., \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Hexadecanoic acid, recognized as an anti-inflammatory fatty acid, also exhibits antifungal and antibacterial activity (Hrichi et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Furthermore, Hexadecanoic acid, 1-pentadecene, and 1-tetradecanol demonstrate antimicrobial activity against \u003cem\u003eAspergillus terreus\u003c/em\u003e, \u003cem\u003eAspergillus niger\u003c/em\u003e, and \u003cem\u003eAspergillus flavus\u003c/em\u003e (Behiry et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). 1,3-dimethyl benzene, also known as m-xylene, can be used as an inert component in non-food pesticide items as per the Environmental Protection Agency (EPA) Chemical and Products Database (CPDat). Mesitylene, also known as 1,3,5-Trimethylbenzene, is commonly used in labs as a unique solvent and frequently used as a reference standard in NMR spectroscopy. Undecane serves as a sex pheromone for certain insects and ants, including Formica lugubris and \u003cem\u003eCaloglyphus rodriguezi\u003c/em\u003e (Walter et al., \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e1993\u003c/span\u003e), and is also a component of the attraction pheromone of \u003cem\u003eOecophylla longinoda\u003c/em\u003e. Additionally, acetaldehyde, a member of the aldehyde group, encompasses a range of highly reactive and hazardous chemicals with diverse industrial applications, including cosmetics, pharmaceuticals, agrochemicals, plastic manufacturing, biofuels, and fragrance production (Aljaafari et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The GC-MS analysis identified several bioactive phytoconstituents with significant biological activities, highlighting their potential applications in various fields, including medicine and industry.\u003c/p\u003e \u003cp\u003eRegarding extraction efficiency, distilled water exhibited the highest extraction efficiency (14.57%), followed by ethanol (9.20%) and ethyl acetate (4.44%), which is consistent with the findings of (Prasathkumar et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Phytochemical analysis of \u003cem\u003eD. metel\u003c/em\u003e demonstrates the presence of various compounds, including flavonoids, saponins, alkaloids, and phenolic substances such as tannins, aligning with the results reported by (Arowora et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eBecause plants co-evolved with the insects that eat them, plants have evolved defense mechanisms. These mechanisms involve the synthesis of substances that disrupt the typical physiological and behavioral features of insects. Consequently, these substances affect feeding, mating, mortality, and egg laying, among other facets of insect life (Adesina, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Natural plant-based essential oils and chemical compounds have a variety of qualities, such as cytotoxicity, antioxidant activity, antifungal and insecticidal, and antibacterial functionality (Sharma et al., \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe LD\u003csub\u003e50\u003c/sub\u003e value of 2.962 mlKg\u003csup\u003e-1\u003c/sup\u003e from \u003cem\u003eD. stramonium\u003c/em\u003e seed extracts was shown by (Jawalkar et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), indicating the potential of seed extract against the rice weevil, S. oryzae. Furthermore, it was observed that \u003cem\u003eD. metel\u003c/em\u003e extract was highly toxic to Fall army worms (FAW) at varying concentrations, such as 60% and 100% in distilled water (Maurawa et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIt is evident that Datura emits a particular scent that has insect-repelling properties.(Acheuk \u0026amp; Doumandji-Mitiche, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) reported similar results, stating that alkaloids disrupt neuroendocrine regulation by deactivating acetylcholinesterase in treated larvae. However, using natural products on crops could be risky if one knows a little bit about their toxicity (Bateman et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIt is clear that Datura emits a particular scent that has insect-repelling properties. According to (Gaire \u0026amp; Subedi, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2013\u003c/span\u003e), the presence of phytochemicals like alkaloids in \u003cem\u003eD. strumonium\u003c/em\u003e extract results in a distinctive smell that serves as a deterrent for a variety of insect pests. (Acheuk \u0026amp; Doumandji-Mitiche, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) reported similar results, stating that alkaloids disrupt neuroendocrine regulation by deactivating acetylcholinesterase in treated larvae. However, using natural products on crops or as personal protective measures could be risky if one knows a little bit about their toxicity (Bateman et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAccording to some earlier research, when \u003cem\u003eD. stramonium\u003c/em\u003e root extract was used at 100% concentration, it showed a distinct larvicidal efficacy ranging from 50\u0026ndash;100% within 24 hours of exposure (Singh et al., \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). (Sharma et al., \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), reported the potential of extracts of the roots, leaves, flowers, and seeds of \u003cem\u003eDatura alba\u003c/em\u003e and \u003cem\u003eDatura stramonium\u003c/em\u003e have significant insecticidal properties.\u003c/p\u003e \u003cp\u003eNumerous studies indicate that plants possess antioxidant attributes primarily due to their phenolic content (Nwozo et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Antioxidants counteract free radicals, prevent oxidative damage, sequester metal ions, and act as oxygen scavengers. Among the solvent extracts, the distilled water extract contained the highest levels of phytochemicals, with total phenolic content measuring 75.03 GAEg⁻\u0026sup1;, followed by ethanol at 17.66 GAEg⁻\u0026sup1; and ethyl acetate at 16.47 GAE g⁻\u0026sup1;. However, the maximum flavonoid content was observed in the ethyl acetate extract (99.32 mgQEg\u003csup\u003e-1\u003c/sup\u003e), followed by dichloromethane (69.86 mgQEg\u003csup\u003e-1\u003c/sup\u003e). Various research studies have shown that alkaloids, phenolic compounds, and flavonoids derived from plants contribute to antioxidant effects (Alam et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), while, phenolic compounds are known for their capacity to inhibit oxidative changes by counteracting free radicals, absorbing oxygen, or decomposing peroxides (Nijveldt et al., 2001). Distilled water proved to be the most effective solvent for extracting phytochemicals from \u003cem\u003eD. metel\u003c/em\u003e, yielding the highest extraction efficiency and total phenolic content, while also demonstrating significant antioxidant properties.\u003c/p\u003e \u003cp\u003eConsequently, the evaluation of phytochemicals is important in phytomedicine, aiding in drug research and advancement (Chihomvu et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). It is important to mention that flavonoids, a type of secondary metabolites, have antibacterial, antioxidant, and anti-aging characteristics and could be useful in cancer treatment by preventing carcinogenesis at different phases. According to Mueed et al., (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), Tannins, which are polyphenols soluble in water, show antimicrobial characteristics. Secondary compounds from plants, such as tannins, phenols, flavonoids, saponins, and essential oils have been shown to possess antioxidant and antimicrobial capabilities, providing defence against harmful infections (Tungmunnithum et al., \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The results showed that each extract exhibited significant DPPH inhibition, with distilled water extract leading with 81.59% inhibition, followed by n-hexane (75.30%), ethanol (66.61%), ethyl acetate (64.06%), and dichloromethane (63.91%). Evaluating phytochemicals, such as flavonoids and tannins, is important for their use in drug development, disease prevention, and treatment due to their strong antioxidant and antimicrobial properties.\u003c/p\u003e \u003cp\u003eHerbal plants containing phenolic compounds and natural antioxidants are employed to counteract oxidative stress by scavenging free radicals (Akbari et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Natural antioxidants are crucial for maintaining good health and lowering the risks associated with chronic diseases by combatting oxidative damage caused by reactive oxygen species (ROS) or environmental oxygen (Sharifi-Rad et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Fruits, vegetables, leaves, and whole grains are common providers of antioxidants like carotenes, phenolic acids, phytoestrogens, vitamin C, and vitamin E, which may lower the risk of disease development (Rahaman et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Our results support the research conducted by Prasathkumar et al., (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), demonstrating the notable antioxidant properties of methanolic extracts from \u003cem\u003eD. metel\u003c/em\u003e leaves, displaying varying degrees of radical scavenging activity between 48.16% and 93.55% at different concentrations, and an IC\u003csub\u003e50\u003c/sub\u003e value of 146.53 \u0026micro;gmL\u003csup\u003e-1\u003c/sup\u003e. Herbal plants, specifically \u003cem\u003eD. metel\u003c/em\u003e, exhibit distinguished antioxidant properties by containing abundant natural antioxidants and phenolic compounds, which can effectively combat oxidative stress and possibly lower the chances of developing diseases. Consequently, \u003cem\u003eD. metel\u003c/em\u003e exhibits promising antioxidant, antimicrobial, and insecticidal activities, highlighting its potential as a source of bioactive phytochemicals. However, further research is required to isolate and identify specific active compounds, and in vivo studies to deepen our understanding of their mechanisms of action as antioxidants and their various biological activities.\u003c/p\u003e "},{"header":"Conclusion","content":"\u003cp\u003eThe present study findings suggest that the leaves of \u003cem\u003eDatura metel\u003c/em\u003e contain a substantial range of chemical compounds, including valuable phytoconstituents such as phenols and flavonoids. Furthermore, GC-MS analysis revealed the presence of significant amounts of these compounds, which are associated with free radical scavenging activity. Similarly, the extracts displayed significant insecticidal activity against \u003cem\u003eB. brassicae\u003c/em\u003e. However, further studies are necessary to isolate and identify these biochemical compounds for potential applications in the pharmaceutical and agrochemical sectors, particularly as alternative medicines and pesticides, respectively.\u003c/p\u003e "},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eContribution of Authors:\u0026nbsp;\u003c/strong\u003eConceptualization, M.J.; Formal analysis, M.A.; A.R. and S.A. Investigation, M.J.; Methodology, M.A.; K.F.A.; A.J.; Project administration, M.J.; Software, M.A.; A.M., and S.F.; Supervision, M.J.; A.E.S.; Visualization, M.A.; A.M., and M.J.; Writing \u0026ndash; original draft, M.A.; Writing \u0026ndash; review \u0026amp; editing, S.R. A.A.; and S.A.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u0026nbsp;\u003c/strong\u003eNational Key Research \u0026amp; Development Program of China (2016YFD0200500) granted research funding for this project. The authors extend their appreciation to Researchers Supporting Project number (RSPD2025R561), King Saud University, Riyadh, Saudi Arabia.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of Interest:\u0026nbsp;\u003c/strong\u003eThere is no conflict of interest by any author.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments:\u0026nbsp;\u003c/strong\u003eProfessor Ji Mingshan provided support \u0026amp; supervision and the experimental guidelines of lab mates of Biopesticides Laboratory, Plant Protection College is greatly acknowledged. The authors extend their appreciation to Researchers Supporting Project number (RSPD2025R561), King Saud University, Riyadh, Saudi Arabia.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAcheuk, F., \u0026amp; Doumandji-Mitiche, B. (2013). Insecticidal activity of alkaloids extract of Pergularia tomentosa (Asclepiadaceae) against fifth instar larvae of Locusta migratoria cinerascens (Fabricius 1781)(Orthoptera: Acrididae). \u003cem\u003eInternational Journal of Science and Advanced Technology\u003c/em\u003e, \u003cem\u003e3\u003c/em\u003e(6), 8\u0026ndash;13.\u003c/li\u003e\n\u003cli\u003eAdesina, J. M. (2022). 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Evaluation of antioxidant activities and total phenolic contents of typical malting barley varieties. \u003cem\u003eFood Chemistry\u003c/em\u003e, \u003cem\u003e107\u003c/em\u003e(1), 296\u0026ndash;304.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Datura metel, GC-MS, Phytochemicals, DPPH, Chemical analysis, Insecticide","lastPublishedDoi":"10.21203/rs.3.rs-6110776/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6110776/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e \u003cem\u003eDatura metel L.\u003c/em\u003e, a plant belonging to the solanaceae family, is renowned for its medicinal, agrochemical and poisonous properties. This research assessed the chemical makeup, antioxidant properties, and phytochemical content of \u003cem\u003eD. metel\u003c/em\u003e leaf extracts by employing different solvents: ethanol (EtOH), ethyl acetate (EA), dichloromethane (DCM), n-hexane (n-Hx), and distilled water (DW). Chemical profile was assessed by Gas Chromatography-Mass Spectrometry (GC-MS) assay whereas; antioxidant activity was determined by using 1, 1-diphenyl-2-picrylhydrazyl (DPPH) assay. While, Insecticidal activity against \u003cem\u003eBrevicoryne brassicae\u003c/em\u003e was evaluated via aphid dip and leaf dip method. On the other hand, quantitative and qualitative phytochemical analysis was carried out by Folin\u0026ndash;Ciocalteu reagent method and aluminum chloride colorimetric method, respectively. GC-MS identified four major compounds as: 11H-Pyrido [3\u0026rsquo;,2\u0026rsquo;:4,5] imidazo [2,1-b] [1,3] benzothiazin-11-one (29.76%), 1,3-dimethyl Benzene (18.35%), Didodecyl phthalate (11.73%), and 1,3,5-Trimethylbenzene (10.09%), along with six minor compounds. Qualitative phytochemical analysis of plant extract discovered alkaloids, flavonoids, phenols, glycosides, and other substances. Whereas, quantitative tests revealed a high concentration of phenols (75.03 GAEg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) in distilled water extracts and increased levels of flavonoids in ethyl acetate (99.33 QEg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e). All samples showed significant antioxidant properties, with DPPH inhibition levels ranging from 63.91\u0026ndash;81.59%. Similarly, \u003cem\u003eD. metel\u003c/em\u003e displayed significant mortality of 81.66% and 73.33% at 48 h exposure at 200 mgmL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e via aphid dip and leaf dip assay, respectively. These findings underline the potential for utilizing \u003cem\u003eD. metel\u003c/em\u003e leaves as a valuable source of bioactive compounds with significant medicinal advantages.\u003c/p\u003e","manuscriptTitle":"Chemical, Insecticidal, Antioxidant and Phytochemical Assessment of Thorn Apple (Datura metel L.) leaf extract from Pakistan's Subtropical Climate","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-04 12:30:27","doi":"10.21203/rs.3.rs-6110776/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":"58ef1f91-09c4-4878-9d23-293bcee2fa04","owner":[],"postedDate":"March 4th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-03-04T12:30:27+00:00","versionOfRecord":[],"versionCreatedAt":"2025-03-04 12:30:27","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6110776","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6110776","identity":"rs-6110776","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}