{"paper_id":"033da2d8-469f-4766-8219-2c7be7b154c9","body_text":"Polyphenolic Bioactive Compounds from Larrea tridentata (DC.) Coville: Extraction, Characterization, Antioxidant, and Antifungal Activities | 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 Polyphenolic Bioactive Compounds from Larrea tridentata (DC.) Coville: Extraction, Characterization, Antioxidant, and Antifungal Activities Muyideen Olaitan Bamidele, Olga B. Álvarez Pérez, José Sandoval-Cortes, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4370220/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 The significance of medicinal plants in inhibiting microbial growth in food and agricultural production as well as their economic viability cannot be overstated. These plants contain secondary metabolites, which are abundant in antimicrobial compounds, such as flavonoids, tannins, saponins, and alkaloids, and their extracts have demonstrated antimicrobial properties against a variety of plant pathogens. The primary objective of this study was to explore the possibility of using bioactive compounds in plant defenses and their biological applications. To achieve this, antifungal polyphenolic bioactive compounds were extracted from the stems and leaves of L. tridentata using conventional methods. The total polyphenol and antioxidant potential of the extracts were assessed and characterized using high-performance liquid chromatography (HPLC). This study compared the polyphenolic constituents of extracts from emerging maceration and Soxhlet extraction techniques in the leaves and stems of Larrea tridentata . The extracts were evaluated for total polyphenolic content (hydrolyzable (HT) and condensed tannins (CT)) and antioxidant activity (ABTS, FRAP, and DPPH). Reverse-Phase High-Performance Liquid Chromatography Electrospray Ionization coupled with mass spectrometry (RP-HPLC-ESI-MS) was used for qualitative identification of antimicrobial phytochemicals. Furthermore, the extracts were analyzed in vitro for antifungal activity against Fusarium oxysporum and Alternaria alternata . The results revealed that 60:40 ethanol:water macerated leaf extract gave the highest hydrolysable tannins (6.41 ± 0.08 mg GAE/g), while its equivalent showed the highest condensed tannins (2.81 mg CE/g). Soxhlet ethyl acetate leaf (SOX ELL) extract showed 1.14 times more condensed tannin content than that of the stems. The antioxidant potential of the extract increased with increasing polarity of the extraction solvent. SOX ELL had higher antifungal effects against F. oxysporum and A. alternata , whereas the 60:40 ethanol: water ratio resulted in 52% inhibition against A. alternata and 43% inhibition against F. oxysporum . Polyphenols with antifungal properties were found in the extracts, including caffeic acid 4-O-glucoside, rhamnetin, protocatechuic acid 4-O-glucoside, kaempferol, (+)-gallocatechin, luteolin, guteolin 7-O-(2-apiosyl-glucoside), gallic acid 4-O-glucoside, cumaric acid 4-O-glucoside, quercetin, NDGA, piceatannol 3-O-glucoside, pterostilbene, tetramethylscutellarein, and cirsimaritin. L. tridentata leaf extracts exhibit potential effectiveness in the development of biological control agents, which can not only enhance crop protection, but also contribute to overall agricultural sustainability. Hydrolysable tannins Condensed tannins Antifungal activity Antioxidants Figures Figure 1 Figure 2 Figure 3 Highlights The antioxidant potential of the extract increased with the polarity of the solvent used in the extraction. The polyphenolic phytochemicals, which are inherent in plants, have been implicated in antifungal activities. The plant composition of hydrolysable and condensed tannins varies with plant species, locations, extraction solvents, extraction methods, and climatic conditions. Plant extracts exhibit potent fungistatic activity that inhibits the growth of polyphagic pathogenic fungi. 1. Introduction Plant fungal infections are a prevalent and economically significant problem in agriculture, horticulture, and natural ecosystems (Traversari et al., 2021 ). These infections can lead to a range of diseases with negative consequences for agricultural output, plant health, and the environment (Fisher et al., 2020 ; Nazarov et al., 2020 ). Bioactive metabolites used in combating fungal diseases are regarded as safe, effective, and environmentally friendly (Devi et al., 2020 ). Bioactive compounds found in plants, microorganisms, and animals have attracted considerable attention because of their wide antimicrobial properties. These compounds include flavonoids, phenolic acids, tannins, carotenoids, sterols, alkaloids and terpenoids (Banwo et al., 2021 ; Echegaray et al., 2023 ; Haruna & Yahaya, 2021 ). They are essential in biological processes, such as antioxidant, antibacterial, anticancer, and antidiabetic activities, as well as in pathogen defense (Almatroodi et al., 2020 ; Bourais et al., 2023 ). Plants produce these bioactive compounds as defense mechanisms in response to threats and stress (Aguirre-Becerra et al., 2021 ; Niazian & Sabbatini, 2021 ). Secondary metabolites such as phenolic compounds are derived from primary metabolites. These compounds are synthesized using precursors, such as amino acids, fatty acids, organic acids, nucleotides, and sugars. (Marchiosi et al., 2020 ; Ranner et al., 2023 ). Secondary metabolites undergo biogenetic processes such as shikimate, polyketide, and mevalonate pathways to form diverse phenolic structures (Fuloria et al., 2022 ; Magray et al., 2023 ). These compounds are characterized by the presence of one or more hydroxyl (–OH) groups attached to a six-carbon aromatic ring (Dikpınar & Süzgeç-Selçuk, 2020 ; Gulcin, 2020 ). Phenolic compounds are present either as unconjugated glycosides or bound to sugars, such as glucose, galactose, rhamnose, xylose, or arabinose in glycosidic forms (González-Sarrías et al., 2020 ). Phenolic acids, flavonoids, tannins, stilbenes, lignans, and coumarins are some examples of phenolic compounds (Mutha et al., 2021 ). They are principally antioxidants because of their electron-donating phenolic groups, and are present in fruits, vegetables, herbs, and medicinal plants (Chiorcea‐Paquim et al., 2020). Polyphenolic compounds are known to provide numerous health benefits and can be extracted using various methods including maceration, Soxhlet extraction, maceration, ultrasound-assisted extraction, microwave-assisted extraction, and fermentation (Jha & Sit, 2022 ; Manousi et al., 2019 ). Each technique has specific advantages and limitations that impact the characteristics and effectiveness of the extracted compounds. Larrea tridentata is a perennial shrub that is native to the semi-desert regions of the United States and Mexico. It is also referred to as the creosote bush, and its extensive medicinal usage dates back through the histories of both nations (Lasché et al., 2023 ; Schwertner-Charão et al., 2022 ). This plant is part of the Zygophyllaceae family, which comprises approximately 30 genera and over 250 species that are predominantly discovered in warm and arid regions (Hadjadj et al., 2022 ; Morales-Ubaldo, Rivero-Perez, et al., 2022 ). L. tridentata typically grows at heights of 0.5 to 3.5 m and produces a light perfume(Bié et al., 2023 ). It has a spreading stem with lanceolate green-yellowish leaves, yellow blooms, and ovoid fruits with tiny white hairs enclosing black seeds(Skouta et al., 2018 ). This plant has gained popularity because to its apparent usefulness in resolving a wide range of health problems(Kannenberg et al., 2022 ). L. tridentata has long been used to treat a variety of health issues, including infertility, rheumatism, arthritis, diabetes, gallstones, kidney stones, common colds, diarrhea, skin problems, obesity, pain, and inflammation. (Morales-Ubaldo, Gonzalez-Cortazar, et al., 2022 ; Torres-León et al., 2023 ). L. tridentata leaves have been found to have strong antifungal activity against various fungal strains, including Rhizoctonia solani . The lanolin extract showed complete inhibition of mycelia at 2000 mg/L, while cocoa butter and water extracts required 3000 and 8000 mg/L for the same inhibition(Castillo et al., 2010 ). L. tridentata leaves were also effective against Pythium spp ., Colletotrichum truncatum , C. coccodes , Alternaria alternata , Fusarium verticillioides , F. solani , F. sambucinum , and Rhizoctonia solani (Osorio et al., 2010 ). According to Chavez-Soliz et al.(2014), aqueous leaf extracts of L. tridentata at concentrations of 1000 mg/L and 5000 mg/L notably reduced the severity of Podosphaera xanthii , which causes powdery melon mildew. Galvan et al. (2014) reported that an aqueous extract of L. tridentata (at concentrations of 10% and 20%) showed significant antifungal effects against Phytophthora capsici and Aspergillus flavus , achieving complete inhibition after 48, 72, and 96 h. Furthermore, the leaf extract of L. tridentata , either alone or in combination with potassium sorbate, effectively inhibited A. flavus growth under pH conditions of 3, 4, and 5, exhibiting a synergistic action when combined. (Munguía et al., 2014). Francisco et al. (2015) assessed the in vitro antifungal activity of L. tridentata water, ethanol, lanolin, and cocoa butter extracts against P. cinnamomi , the ethanol extract completely inhibited mycelium, while the lanolin extract showed very low inhibition. L. tridentata ethanol and dichloromethane extracts reduced fungal growth by 75–100% against A. tenuissima , A. niger , Penicillium polonicum , and Rhizopus oryzae . Aguirre-Joya et al. ( 2018 ) used bioactive films comprising L. tridentata polyphenols to achieve 50% inhibition of A. alternata , F. oxysporum , Botrytis cinerea , and C. gloeosporioides . This study addressed the potential of bioactive compounds in plant defense and their biological applications by extracting antifungal polyphenolic bioactive compounds from L. tridentata stems and leaves through maceration and Soxhlet extraction, assessing the total polyphenol content and antioxidant potential, and characterizing the extracts using high-performance liquid chromatography (HPLC). 2. Materials and Methods 2.1 Chemicals and reagents Analytical grade reagents, including anhydrous sodium carbonate (Na 2 CO 3 ), sodium hydroxide (NaOH), sodium carbonate (Na 2 CO 3 ), sodium nitrite (NaNO 2 ), aluminum chloride (AlCl 3 ), sodium hydroxide (NaOH), hydrochloric acid (HCl), and ammonia ferric sulphate ([FeNH 4 (SO 4 ) 2 O.12 H 2 O]), were supplied by Sigma Aldrich (Toluca, México). Other chemicals also purchased included ethanol, distilled water, potato dextrose agar, gallic acid, quercetin, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2′-Azino-bis (3-ethylbenzthiazoline-6-sulphonic acid, (ABTS) 2,4,6-Tripyridyl-s-triazine (TPTZ), Tween 80, and Folin-Ciocalteu reagent. 2.2 Plant materials The plant material, encompassing leaves and stems, was procured in February 2023 at coordinates Latitude 25 o 10′59.3′′ and Longitude 102 o 45′41.2′ in Parras de la Fuente, Coahuila, Mexico, by Dr. Cristian Torres, a researcher at the Research Center and Ethnobiological Garden (CIJE), Universidad Autonoma de Coahuila, Unidad Torreón, Viesca, Coahuila. The plants were manual defoliation, the branches were washed with distilled water and air-dried in the laboratory for 14 d. Subsequently, the leaves and stems pulverised using an automatic grinder at the Department of Biotechnology, Faculty of Chemical Science, Autonomous University of Coahuila, Mexico. The resulting pulverized samples were stored in plastic bags in the dark at 4°C until needed. 2.3 Extraction of polyphenolic compounds 2.3.1 Maceration extraction Pulverized samples of L. tridentata leaves and stems were extracted by maceration. 5 g of sample was dissolved in 100 mL of water at a solid-to-liquid ratio of 1:10 (w/v) in the shaker. The extraction was performed for 24 h at 30°C with constant agitation at 200 rpm in a shaker. The process was repeated using mixture of water-to-ethanol ratios, namely 50:50, 40:60, and 0:100 (v/v) ethanol: water ratio. 2.3.2 Soxhlet extraction The extraction equipment included a distillation round flask, a glass condensed linked to 4°C water, a thimble for sample placement, and an electric heat system. The solid-to-liquid ratio is 1:5 (w/v). A total of 5 h was spent extracting 50 g of material in 250 mL of solvent. The solvents were selected in the order of their polarity differences: n -hexane, ethyl acetate and diethyl ether. After recovering the extracts, the solvent was removed under vacuum and kept at 20°C for future analysis to prevent degradation. 2.4 Total Polyphenolic Constituents of extracts The total polyphenolic components in each extract were determined using the technique described by Georgé et al. (2005) for hydrolysable and condensed tannins methodology proposed by Amaya-Chantaca et al. (2021). The total polyphenolic compounds per L. tridentata (mg TPC/g L. tridentata ) were reported. 2.4.1 Hydrolysable tannins (Folin Ciocalteu) The total polyphenolic compounds were determined using the Folin Ciocalteu reagent, requiring only minor changes to the approach provided by Georgé et al. (2005). The Folin Ciocalteu test is an electron transfer technique that provides reducing capability, which is represented as phenolic content. To begin, 25 µL of diluted sample, 25 µL of Folin Ciocalteu reagent, and 25 µL of sodium carbonate (0.7 M) were mixed. After 5 min, 125 µL of distilled H 2 O was added and the sample was measured at 750 nm (UV-visible Epoch TM Microplate Spectrophotometer). According to linear regression using a calibration curve, the results were presented as milligrams of gallic acid equivalents per milli gram of L. tridentata (mg GAE/ g of L. tridentata ). 2.4.2 Condensed tannins (HCl-Butanol) Condensed tannins were quantified using the method described by Amaya-Chantaca et al. (2021) with a modification proposed by Swain and Hillis (1959). 250 µL of water were combined with 1.5 mL of HCl: Butanol (1:9) and 1 mL of ferric reagent. The reaction mixture was heated to 95°C for 45 min. The mixture is allowed to cool for 30 min before being read at 460 nm with a UV-visible spectrophotometer (UV-visible Epoch TM Microplate Spectrophotometer). According to linear regression using a calibration curve, the results were expressed as catechin g equivalents per gram (mg CE/g). 2.5 Antioxidant activity in the extracts 2.5.1 DPPH radical scavenging assay The methodology was carried out with minor modifications according to the method proposed by Castro-Lopez et al. (2019); the electron donation capacity of the samples was evaluated from a purple colour solution of radical DPPH, using methanol as solvent (60 mM). Subsequently, 193 µL of DPPH radical were added to each microplate for every 7 µL of sample or standard curve (gallic acid). The reaction solution was subjected to an incubation period in the dark for 30 min; subsequently, the absorbance of the samples was recorded at a wavelength of 517 nm (UV–visible Epoch ™ Microplate Spectrophotometer), and Trolox was used as a standard, and the results were expressed as mg Trolox equivalents per gram (mg TEAC /g) 2.5.2 Ferric reducing ability (FRAP) The ferric reducing ability was determined according to the method of Delgado-Andrade et al. (2005) with some modifications. The FRAP reagent was prepared by 2.5 mL of a 10 mM TPTZ solution in 40 mM HCl plus 2.5 mL of 20 mM FeCl 3 ‚ H 2 O and 2.5 mL of 0.3 M acetate buffer (pH 3.6). A 290 µL of FRAP reagent was mixed with 10 µL of the sample or standard concentration (Trolox). The reaction mix was incubated in darkness for 15 min and read by UV–visible Epoch ™ Microplate Spectrophotometer automatic sample positioner (593 nm); the results were also expressed as milligrams equivalents of Trolox per gram of sample (mg TEAC/g). 2.5.3 Radical Scavenging Activity ABTS A 7 mM ABTS solution was prepared, mixed with 2.45 mM K 2 S 2 O 8 , and left to incubate at room temperature for 12–16 h in the dark. The solution was diluted with ethanol until obtaining an absorbance of 0.7 ± 0.02 at 734 nm. Then, 190 µL of ABTS ethanolic solution and 10 µL of the extracts were mixed, allowed to react for 1 min, and read at 734 nm in a microplate reader (Epoch, Biotek, Instruments, Inc.). The ABTS solution with the solvent of the samples was taken as a control. Trolox was used as a standard, and the results were expressed as mg Trolox equivalents per gram of extract (mg TEAC/g). 2.6 Antifungal assay of the extracts The antifungal activities of the extracts were assayed against F. oxysporum and A. alternata using the modified method of Eisa et al. (2017). Molten potato dextrose agar medium was poisoned with 0.5 mL of extracts mixed under aseptic conditions in laminar flow, and the medium was allowed to solidify. Each plate was inoculated with F. oxysporum and A. alternata using a 5 mm thick disc of fungus (spores and mycelium) and incubated at 25 ± 1°C until fungal growth in the control Petri plates was completed. The solvents used for the extraction were used as negative controls. The plates were incubated and monitored for 5–8 days to confirm the complete growth of the fungal. Each test was performed in triplicate by measuring the relative growth of the fungus in the treatment versus the control. The fungal mycelial growth (mm) in treated (T) and control (C) conditions was measured diametrically in three different directions, and the percentage inhibition in radial growth (I) was calculated using the Eq. 1 (Vincent, 1947). \\(I=\\frac{(C-T)}{C}*100\\) Eq. 1 where I = Inhibition of mycelia growth; C = Growth of fungus in control, and T = Growth of fungus in treatment. 2.7 Reverse phase high-performance liquid chromatography RP-HPLC-ESI-MS analysis of extracts Reverse phase high-performance liquid chromatography (HPLC) analyses were conducted using a Varian HPLC system, which included the following components: an autosampler (Varian ProStar 410, USA), a ternary pump (Varian ProStar 230I, USA), and a PDA detector (Varian ProStar 330, USA). Additionally, a liquid chromatograph ion trap mass spectrometer (Varian 500-MS IT Mass Spectrometer, USA) equipped with an electrospray ion source was used. The samples (5 µL) were introduced into a Denali C18 column (150 mm × 2.1 mm, 3µm, Grace, USA). The column temperature was maintained at 30°C. Formic acid (0.2% v/v; solvent A) and acetonitrile (solvent B) were used as elution solvents. The following gradient program was applied: an initial composition of 3% B, linear increase to 9% B from 0 to 5 min, linear increase to 16% B from 5 to 15 min, and final linear increase to 50% B from 15 to 45 min. Subsequently, the columns were cleaned and conditioned. The flow rate was held constant at 0.2 mL/min, and detection was performed at wavelengths of 245, 280, 320, and 550 nm. The entire effluent, at a rate of 0.2 mL/min, was introduced into the mass spectrometer without splitting. All mass spectrometry (MS) experiments were performed in negative ionization mode ([M-H] −1 ). Nitrogen served as the nebulizing gas and helium functioned as the damping gas. Specific ion source parameters included a spray voltage of 5.0 kV, capillary voltage of 90.0 V, and temperature of 350°C. Data collection and processing were performed using MS Workstation software (Version 6.9). Samples were initially analyzed in full scan mode across a mass-to-charge ratio (m/z) range of 50–2000. Phenolic compounds were identified by comparing the HPLC retention times with the MS data. Molecular weights were cross-referenced with literature and a database maintained by the Departamento de Investigación en Alimentos at the Universidad Autónoma de Coahuila (DIA-UAdeC). 2.8 Experimental design and statistical analysis The experiments were carried out at three different measurement levels, and the outcomes are presented as the average value along with the standard deviation (SD). Data analysis involved the use of one-way analysis of variance (ANOVA), with the factor being the combination of cultivar and location. Statistical analysis was performed using the IBM SPSS Statistics software (SPPS Software). 3. Results and discussion 3.1 Polyphenol composition (hydrolysable tannins) Tannin may be found in the wood, stems, barks, leaves, roots, and fruits of any plant species, and its quantity varies by plant component. Tannin-rich plants are the most often utilized raw material for industrial-level tannin extraction (Zhang et al., 2023). Hydrolysable tannins offer a wide range of biological activities, including antioxidant, anti-inflammatory, and antifungal characteristics. This study revealed that the hydrolysable tannin content of macerated leaves of L. tridentata ranges from 2.72–6.41 mg GAE/g and the macerated stem extracts ranged from 0.93–1.64 mg GAE/g (Fig. 1 (I)). The highest hydrolysable tannins were recorded with 60:40 ethanol: water 6.41 ± 0.08 mg GAE/g (Fig. 1 (I)). The macerated stem aqueous extract showed the lowest hydrolysable content. The macerated leaf extracts were 3.91 times more hydrolysable than the stems of L. tridentata . Plant leaves often contain a larger quantity of hydrolysable tannins than stems, roots, and flowers (Dettlaff et al., 2018 ). Hydrolysable tannins are polyphenolic compounds that are present in various plant species. These tannins frequently act as plant defense mechanisms against herbivores and diseases, and they are more abundant in particular plant sections, with leaves being a typical reservoir for them (Savina et al., 2023 ). The ethyl acetate leaf extract of L. tridentata contained the highest levels of hydrolysable tannins (Fig. 1 (II)). This was followed by ethyl acetate and diethyl ether stem extracts. The extraction method and solvent polarity are among the factors that determine the composition of the extract (Akinmoladun et al., 2022 ; Jasso de Rodríguez et al., 2019). This result confirmed the effect of polarity on the composition of the extracts. The soxhlet ethyl acetate leaf extract (Sox ELL) contained 13.5 times hydrolysable tannins than the n -hexane leaf extracts (Fig. 1 (II)). There is a possibility of bonding between the oxygen atom of ethyl acetate and hydrogen of the hydroxyl of the tannins leading to higher composition of tannins (Sulistiyani et al., 2022 ). The SOX ELL contained the highest hydrolysable tannins 6.37 ± 0.06 mg GAE/g of leaves L. tridentata . This study obtained values of hydrolysable tannins higher than the work reported by Cadena et al. ( 2023 ). The phytochemical profile of a plant is influenced by genetics, environmental conditions, cultivation practices, harvesting time, post-harvest processing methods, plant parts, storage conditions, and biotic and abiotic stressors (Guan et al., 2021 ; Liebelt et al., 2019 ). 3.2 Condensed tannins The composition of condensed tannins in the macerated leaves and stems extracted from L. tridentata is shown in Fig. 2 . The values of condensed tannins vary from 0.75–3.29 CE/g in the leaves and 0.86–2.81 mg CE/ g in the stems (Fig. 2 (I&II). The leaves contained 1.17 times more condensed tannins in the leaves than stems. 60:40 ethanolic macerated leaves extracts gave the highest condensed tannins composition (3.30 ± 0.012 mg CE/g). While also it equivalent in stems gave the best composition of condensed tannins (2.81 mg CE/g, Fig. 2 (I)). The plant composition of tannins varies on plant species and climatic circumstances(Bule et al., 2022). The aqueous extract of macerated stems contained more condensed tannins than hydrolysable tannins. Although, the concentration of condensed tannins were found throughout L. tridentata in low concentrations(Hyder et al., 2002 ). The condensed tannin content in soxhlet leaf extracts ranged from 0.96 to 3.32 mg CE/g, while the stems exhibited values between 0.69 and 2.90 mg CE/g (as shown in Fig. 2 (II)). This indicates that the condensed tannin content in the leaves is higher than that in the macerated extracts. The use of polar organic solvents can enhance bonding with phytoconstituents, leading to an increased tannin content. The leaves had a condensed tannin content that was 1.14 times greater than that of the stems. Among the leaf extracts, the ethyl acetate extract contained the highest condensed tannin content compared to the n -hexane extract. This higher content in the ethyl acetate extract can be attributed to the polarity of the solvent used, as the hydroxyl hydrogen of condensed tannins can interact with the oxygen atom in ethyl acetate, thereby increasing tannin quantity. The condensed tannin content tends to increase as the polarity of the solvent used for extraction increases (Ng et al., 2020 ) . 3.3 Antioxidant Activity Plant extracts are rich in antioxidants including phenolic compounds, carotenoids, vitamins, and polyphenols. These compounds can scavenge free radicals, neutralize metal ions, and play a crucial role in preventing and controlling diseases caused by pathogens such as fungi, bacteria, and viruses. The antioxidant activity of plants is largely attributable to the presence of bioactive compounds. The antiradical action is primarily related to the polyphenolic content, and as the polarity of the solvent increases, the number of hydroxyl groups in the extract medium increases, as does the probability of hydrogen donation to free radicals(Albert et al., 2022 ; Bautista-Hernández et al., 2022 ; Hodhodi et al., 2022 ). The DPPH assay is routinely used to evaluate the ability of antioxidants to scavenge free radicals. This involves quick electron transfer followed by delayed hydrogen transfer. The effectiveness of the hydrogen transfer process can be affected by the solvent used, particularly those capable of receiving hydrogen bonds such as ethanol or methanol (Isa et al., 2018 ; Tymczewska et al., 2023 ). The extract with the highest scavenging potential was the macerated leaf extract of L. tridentata extracted with ethanol: water, 60:40 (191.48 ± 3.37 mg TEAC/g), whereas the ethanolic stem extract showed the highest scavenging potential (166.29 ± 1.69 mg TEAC/g) for the macerated stem extract. The macerated water extract showed the lowest antiradical activity in both leaves and stems of L. tridentata . SOX ELL extracts showed the highest scavenging activity (120.74 ± 7.14 mg TEAC/g), whereas SOX HLS showed the lowest antioxidant activity (35.67 ± 1.11 mg TEAC/g). The polarity of the solvent plays a crucial role in determining the antioxidant activity and extraction yield of the phytochemicals(Jeyaraj et al., 2021 ; Truong et al., 2019 ). Polar solvents tend to extract more polar components, which often possess stronger antioxidant properties, whereas nonpolar solvents tend to extract less polar components with weaker antioxidant activity(Lohvina et al., 2021 ; Wang et al., 2023 ). However, the leaves of L. tridentata possessed a higher antioxidant potential than the stems. Antioxidants provided by L. tridentata could serve as free radical scavengers and mitigate Reactive Oxygen Species /free radicals or could contribute to preventing the formation of hydroxyl radicals by deactivating free metal ions through chelation or converting H 2 O 2 to other innocuous compounds (such as water and oxygen). The ABTS radical-scavenging activity of the extracts showed a trend like that of DPPH. However, the value of the ABTS result of the antioxidant potential of the extracts was higher than that of the DPPH. This could be attributed to the incubation time (30 min) used in the assay. Skouta et al., ( 2018 ) reported decreases in the DPPH scavenging ability with increase in incubation time. The 60:40 ethanol: water macerated leaf extract of L. tridenatata (225.57 ± 4.51 mg TEAC/g) showed the highest antioxidant activity compared to the water macerated leaf extract (106.07 ± 5.97 mg TEAC/g). The macerated stem extract showed the highest antiradical activity at 50:50 ethanol: water (117.07 ± 3.40 mg TEAC/g). The ELL extract showed the highest antiradical activity (226.57 ± 1.04 mg TEAC/g) compared to the n-hexane soxhlet extract of leaf and stem, with antioxidant activity of (55.07 ± 5.20 mg TEAC/g) and (43.57 ± 1.26 mg TEAC/g) respectively. The FRAP test is a useful technique for determining the antioxidant capacities of diverse compounds. The reductive ability of the extract is consistent with that reported by (Skouta et al., 2018 ) with 60:40 ethanol: water having the highest reductive property (128.95 ± 1.09 mg TEAC/g). The n -hexane extracts had fewer polyphenolic constituents and accounted for their low antioxidant potential. The polarity of the solvent determines the phytochemicals inherent in the extract and accounts for the higher antioxidant potential of the extract with higher solvent polarity. Table 1 The antioxidant activities of extracts obtained from both maceration and Soxhlet extraction of leaves and stems of Larrea tridentata . The values are reported as mean value with standard deviation. Method Of Extraction And Plant Part Extract DPPH (mg TEAC/g) ABTS (mg TEAC/g) FRAP (mg TEAC/g) MLL Aqueous 43.3 ± 6.19 j 106.1 ± 5.97 j 58.7 ± 0.72 h 50% ET 106.7 ± 5.88 f 168.9 ± 5.07 e 75.4 ± 0.94 f 60% ET 191.5 ± 3.57 a 225.6 ± 4.51 b 128.9 ± 1.09 c 100% ET 151.9 ± 0.64 d 209.7 ± 2.84 c 87.5 ± 0.79 e MLS Aqueous 31.1 ± 2.94 m 80.6 ± 2.02 l 23.0 ± 1.09 l 50% ET 106.7 ± 2.64 f 117.1 ± 3.40 g 62.5 ± 0.09 g 60% ET 152.9 ± 1.69 c 113.6 ± 3.33 h 117.0 ± 0.48 d 100% ET 166.3 ± 1.69 b 93.2 ± 2.02 k 15.0 ± 0.94 m SOXHLET ELL 120.7 ± 7.14 e 226.6 ± 1.04 a 55.1 ± 1.25 i ELS 103.3 ± 1.11 g 195.9 ± 5.01 d 21.9 ± 0.63 l HLL 36.7 ± 6.94 k 55.1 ± 5.20 m 49.3 ± 0.48 j HLS 35.7 ± 1.11 l 43.6 ± 1.26 n 34.6 ± 0.48 k DLL 85.9 ± 1.69 i 154.9 ± 1.50 f 150.5 ± 1.25 a DLS 88.9 ± 2.22 h 111.7 ± 3.82 i 144.2 ± 0.48 b Macerated L. tridentata leaves (MLL), macerated L. tridentata stems (MLS), Soxhlet Ethyl acetate L. tridentata stems (ELS), Soxhlet Ethyl acetate L. tridentata leaves (ELS), Soxhlet n -hexane L. tridentata leaves (Sox HLL), Soxhlet n -hexane L. tridentata stems (Sox HLS), Soxhlet Diethyl ether L. tridentata leaves (Sox DLL) and Soxhlet Diethyl ether L. tridentata leaves (Sox DLL). 3.4 Antifungal activities Plant extracts with high fungistatic activity inhibit the growth of polyphagic pathogenic fungi, demonstrating their potential as natural antifungal agents(Kursa et al., 2022 ). Table 2 , Fig. 3 : shows the effects of L. tridentata extract on the mycelial growth of A. alternata and F. oxysporum . At 5% concentration, the SOX ELL extract inhibited A. alternata by 79.12 ± 0.014% and F. oxysporum by 80.71 ± 0.38% (Table 2 , Fig. 3 ). The SOX ELS extract at 5% inhibited the mycelial growth of A. alternata by 61.57 ± 0.042% and F. oxysporum by 70.59 ± 0.308%. Similarly, SOX DLL at 5% inhibited the mycelial growth of A. alternata by 64.42 ± 0.16% and F. oxysporum by 67.36 ± 1.00%. The 5% SOX DLS extract inhibited A. alternata by 57.49 ± 0.27% and F. oxysporum by 65.60 ± 0.17%. The 5%, 50:50 ethanol: water macerated leaf extract inhibited the mycelial growth of A. alternata by 42.42 ± 0.58% and F. oxysporum by 50.03 ± 0.39%. In addition, 5%, 60:40 ethanol: water macerated leaf extract inhibited the mycelial growth of phytopathogens by 52.16 ± 0.014% and 42.52 ± 0.189%, respectively. However, the macerated stem extracts showed lower inhibition, ranging from 13–41% for A. alternata and 0.04–15% for F. oxysporum . These results suggest that extracts with higher polyphenol content, such as SOX ELL, exhibited stronger fungicidal activity against A. alternata and F. oxysporum . Additionally, the leaves generally showed higher inhibition than the stems, likely because of the higher polyphenol levels in the leaves. Conversely, aqueous extracts display low resistance to phytopathogens because of their low polyphenol content and limited ability to extract bioactive phytochemical compounds, possibly extracting more primary metabolites such as sugars. Liu et al., 2021 reported low inhibition of aqueous extract on Aspergillus flavus . Guo et al., 2023, reported that fungicides with inhibition percentages between 75% and 90% are sensitive to phytopathogens and exhibit good fungicide efficiency. Table 2 The Antifungal activities of macerated and Soxhlet extracts of Leaves and stems of L. tridentata Method Of Extraction and Plant Part Extracts % Inhibition of ( A. Alternata ) % Inhibition of ( F. Oxysporum ) MLL AQUEOUS 19.5 ± 0.31 i 11.2 ± 0.52 l 50% ET 42.4 ± 0.58 f 50.0 ± 0.39 e 60% ET 52.2 ± 0.01 e 42.5 ± 0.19 f 100% ET 25.5 ± 0.27 h 20.9 ± 0.04 i MLS AQUEOUS 13.2 ± 0.07 k 0.04 ± 0.03 n 50% ET 41.1 ± 0.07 g 12.8 ± 0.28 k 60% ET 25.6 ± 0.03 h 9.9 ± 0.26 m 100% ET 17.4 ± 0.79 j 15.2 ± 0.72 j SOXHLET ELL 79.1 ± 0.01 a 80.7 ± 0.38 a ELS 61.6 ± 0.04 c 70.6 ± 0.31 b HLL 17.7 ± 1.59 j 36.8 ± 1.25 g HLS 9.4 ± 0.77 l 24.20 ± 0.59 h DLL 64.4 ± 0.16 b 67.36 ± 1.00 c DLS 57.5 ± 0.27 d 65.60 ± 0.17 d 3.5 Reverse phase high-performance liquid chromatography (HPLC) Reverse-phase high-performance liquid chromatography (HPLC) of macerated and Soxhlet leaves and stems of L. tridentata revealed the presence of twenty-five (25) compounds (Table 3 ), while twenty-two compounds were recorded for the macerated and Soxhlet stem extracts (Table 4 ). The result revealed that most polyphenolic phytochemical and inherent in the leaves of L. tridentata . Which can be implicated for its antifungal effects. The class of compounds varies from lignans to catechins, acids, flavonols, and flavones. Fifteen (15) of these compounds were polyphenols, including caffeic acid 4-O-glucoside, rhamnetin, protocatechuic acid 4-O-glucoside, kaempferol, (+)-gallocatechin, luteolin, guteolin 7-O-(2-apiosyl-glucoside), gallic acid 4-O-glucoside-Coumaric acid 4-O-glucoside, quercetin, dihydroquercetin, piceatannol 3-O-glucoside, pterostilbene, tetramethylscutellarein, and cirsimaritin. Among the phenolic compounds of L. tridentata , a ligann such as NDGA stands out since it is extracted from the resin in leaves and stems and have been attributed to the biological activities of plants, particularly L. tridentata (Lira et al., 2003). NDGA has been shown to inhibit several phytopathogenic fungi, and its potential mechanism of action has been explored. This phytochemical may decrease the growth and persistence of fungal colonies by interfering with the production of fungal biofilms. NDGA inhibits the formation of ergosterol, a critical component of fungal cell membranes. This disruption affects membrane integrity, resulting in decreased fungal growth and viability. The antifungal activity of NDGA is also attributed to its ability to inhibit microbial growth and cell function. Li et al. ( 2023 ) suggested that NDGA may inhibit the synthesis of pro-inflammatory molecules such as Tumor Necrosis Factor-alpha (TNF-α), which may contribute to its antifungal action. Furthermore, NDGA's comprehensive strategy, which targets both membrane integrity and inflammatory pathways, makes it a good option for antifungal applications. Table 3 The Reverse phase high-performance liquid chromatography (HPLC) analyses of macerated and soxhlet leaves extrcats of L. tridentata Compound (M-H)- Class MLL H 2 O MLL 50% MLL 60% MLL 100% Et SOX ELL SOX DLL SOX HLL Caffeic acid 4-O-glucoside 340.9 Hydroxycinnamic acids X X X 5-Heptadecylresorcinol 346.9 Alkylphenols X X X X X Rhamnetin 315 Methoxyflavonols X X X X X 3,7-Dimethylquercetin 328.9 Methoxyflavonols X X X X X Protocatechuic acid 4-O-glucoside 316.9 Hydroxybenzoic acids X X X Methylgalangin 284 Methoxyflavonols X X X Glycitein 283.9 Methoxyisoflavones X X X Kaempferol 298 Methoxyflavonols X X X Nordihydroguaiaretic acid (NDGA) 301 Lignans X X X X (+)-Gallocatechin 305.1 Catechins X X X 3-p-Coumaroylquinic acid 336.8 Hydroxycinnamic acids X Luteolin 285 Flavones X X X X X Luteolin 7-O-(2-apiosyl-glucoside) 580.9 Flavones X X Methylgalangin 284.9 Methoxyflavonols X X X 3-p-Coumaroylquinic acid 336.8 Hydroxycinnamic acids X X X Secoisolariciresinol 364.9 Lignans X Gallic acid 4-O-glucoside 332.9 Hydroxybenzoic acids X X X Avenanthramide 2c 314.9 Hydroxycinnamic acids X X X p-Coumaric acid 4-O-glucoside 325 Hydroxycinnamic acids Dihydroquercetin 304 Dihydroflavonols X X X X X Quercetin 300.9 Flavonols X Piceatannol 3-O-glucoside 405 Stilbenes X X Pterostilbene 254.9 Stilbenes X X X X X Tetramethylscutellarein 340.9 Methoxyflavones X X Cirsimaritin 312.9 X X - Present Table 4 The reverse phase high-performance liquid chromatography (HPLC) analyses of macerated and soxhlet stems extracts of L. tridentata. Compound (M-H)- Class MLS H2O MLS 50% MLS 60% MLS 50% SOX ELS SOX DLS SOX HLS Caffeic acid 4-O-glucoside 340.9 Hydroxycinnamic acids X X X 5-Heptadecylresorcinol 346.9 Alkylphenols X X X X Rhamnetin 315 Methoxyflavonols X X X 3,7-Dimethylquercetin 328.9 Methoxyflavonols X X Protocatechuic acid 4-O-glucoside 316.9 Hydroxybenzoic acids X X X Methylgalangin 284 Methoxyflavonols X X Glycitein 283.9 Methoxyisoflavones X X Kaempferol 298 Methoxyflavonols X X NDGA 301 Lignans X X X X (+)-Gallocatechin 305.1 Catechins X X X 3-p-Coumaroylquinic acid 336.8 Hydroxycinnamic acids X Luteolin 285 Flavones X X X X Luteolin 7-O-(2-apiosyl-glucoside) 580.9 Flavones X X Methylgalangin 284.9 Methoxyflavonols X X X 3-p-Coumaroylquinic acid 336.8 Hydroxycinnamic acids X X X Secoisolariciresinol 364.9 Lignans X Gallic acid 4-O-glucoside 332.9 Hydroxybenzoic acids X Avenanthramide 2c 314.9 Hydroxycinnamic acids X X X p-Coumaric acid 4-O-glucoside 325 Hydroxycinnamic acids X X X Dihydroquercetin 304 Dihydroflavonols X X Quercetin 300.9 Flavonols X X X Tetramethylscutellarein 340.9 Methoxyflavones X X X - Present 4. CONCLUSION The findings imply that L. tridentata extracts have the potential to be employed in the control of plant diseases caused by A. alternata and F. oxysporum . Notably, SOX ELL and SOX DLL extracts at 5% concentration revealed consistent effectiveness in lowering the frequency and severity of these plant diseases, consequently preventing their growth, and alleviating their negative effects on plants. To our knowledge, this is the first in vitro evaluation of the polyphenol content, antioxidant potential, and antifungal activity of L. tridentata stems and leaves extracts. However, it is recommended that this study be expanded to the commercial greenhouse level by utilizing more ecologically friendly solvents. Abbreviations HT-Hydrolysable tannins CT-condensed tannins ABTS- 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) FRAP- (Ferric Reducing Antioxidant Power) DPPH - (2,2-diphenyl-1-picrylhydrazyl) HPLC - High-Performance Liquid Chromatography RP-HPLC-ESI-MS - Reverse-Phase High-Performance Liquid Chromatography-Electrospray Ionization-Mass Spectrometry NDGA - Nordihydroguaiaretic acid TPTZ - 2,4,6-Tris(2-pyridyl)-s-triazine, GAE/g - Gallic Acid Equivalents per gram SOX ELS - Soxhlet Ethyl acetate extract of L. tridentata stems SOX ELL - Soxhlet Ethyl acetate extract of L. tridentata leaves Sox HLL - Soxhlet n-hexane extract of L. tridentata leaves Sox HLS - Soxhlet n-hexane extract of L. tridentata stems Sox DLL - Soxhlet Diethyl ether extract of L. tridentata leaves Sox DLS - Soxhlet Diethyl ether extract of L. tridentata stems CE/g - Condensed Tannin Equivalents per gram TEAC/g - Trolox Equivalent Antioxidant Capacity per gram Declarations CRediT authorship contribution statement Muyideen O. Bamidele: Conceptualization, Formal analysis, Investigation, Methodology, Writing – original draft. Olga B. Álvarez Pérez: Data curation, Resources, Visualization, Methodology, Supervision. José C. Sandoval: Data curation, Formal analysis, Methodology, Visualization, Supervision, Writing – review & editing. María L. Flores-López: Formal analysis, Methodology, Supervision, Writing – original draft. Mónica L. Chavez-González: Methodology, Formal analysis,Supervision, Writing – review & editing. Cristóbal N. Aguilar: Supervision, Data curation, Resources, Methodology, Formal analysis, Conceptualization, Writing – review & editing, Writing – original draft. Acknowledgements Muyideen Olaitan Bamidele appreciated National Council of Humanities, Sciences and Technologies (CONACYT, Mexico) for PhD fellowship support awarded to me (1245491). Conflict of interest The authors declare no conflict of interest. References Aguirre-Becerra, H., Vazquez-Hernandez, M. 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(2022). nor 3′-Demethoxyisoguaiacin from Larrea tridentata Is a Potential Alternative against Multidrug-Resistant Bacteria Associated with Bovine Mastitis. Molecules , 27 (11). https://doi.org/10.3390/molecules27113620 Morales-Ubaldo, A. L., Rivero-Perez, N., Valladares-Carranza, B., Madariaga-Navarrete, A., Higuera-Piedrahita, R. I., Delgadillo-Ruiz, L., Bañuelos-Valenzuela, R., & Zaragoza-Bastida, A. (2022). Phytochemical Compounds and Pharmacological Properties of Larrea tridentata. Molecules , 27 (17), 5393. https://doi.org/10.3390/molecules27175393 Munguía, A. R., Zárate, M. A., Luisa, M., Inungaray, C., Valles, C., & Mx, M. M. (n.d.). Efecto del uso combinado de extracto de Larrea tridentata y sorbato de potasio sobre el crecimiento de Aspergillus flavus . www.reibci.org Mutha, R. E., Tatiya, A. U., & Surana, S. J. (2021). Flavonoids as natural phenolic compounds and their role in therapeutics: an overview. Future Journal of Pharmaceutical Sciences , 7 (1), 25. https://doi.org/10.1186/s43094-020-00161-8 Nazarov, P. A., Baleev, D. N., Ivanova, M. I., Sokolova, L. M., & Karakozova, M. V. (2020). Infectious plant diseases: etiology, current status, problems and prospects in plant protection. Acta Naturae , 12 (3), 46–59. https://doi.org/10.32607/actanaturae.11026 Ng, Z. X., Samsuri, S. N., & Yong, P. H. (2020). The antioxidant index and chemometric analysis of tannin, flavonoid, and total phenolic extracted from medicinal plant foods with the solvents of different polarities. Journal of Food Processing and Preservation , 44 (9). https://doi.org/10.1111/jfpp.14680 Niazian, M., & Sabbatini, P. (2021). Traditional in vitro strategies for sustainable production of bioactive compounds and manipulation of metabolomic profile in medicinal, aromatic and ornamental plants. Planta , 254 (6), 111. https://doi.org/10.1007/s00425-021-03771-5 Osorio, E., Flores, M., Hernández, D., Ventura, J., Rodríguez, R., & Aguilar, C. N. (2010). Biological efficiency of polyphenolic extracts from pecan nuts shell (Carya Illinoensis), pomegranate husk (Punica granatum) and creosote bush leaves (Larrea tridentata Cov.) against plant pathogenic fungi. Industrial Crops and Products , 31 (1), 153–157. https://doi.org/10.1016/j.indcrop.2009.09.017 Ranner, J. L., Schalk, S., Martyniak, C., Parniske, M., Gutjahr, C., Stark, T. D., & Dawid, C. (2023). Primary and Secondary Metabolites in Lotus japonicus . Journal of Agricultural and Food Chemistry , 71 (30), 11277–11303. https://doi.org/10.1021/acs.jafc.3c02709 Savina, T., Lisun, V., Feduraev, P., & Skrypnik, L. (2023). Variation in Phenolic Compounds, Antioxidant and Antibacterial Activities of Extracts from Different Plant Organs of Meadowsweet (Filipendula ulmaria (L.) Maxim.). Molecules , 28 (8), 3512. https://doi.org/10.3390/molecules28083512 Schwertner-Charão, L., Delgado-Martínez, R., Treviño-Carreón, J., Jiménez-Sierra, C. L., Astudillo-Sánchez, C. C., & Osorio-Hernández, E. (2022). Interactions between facilitator species and Lophophora williamsii (Lem. ex Salm-Dyck) J.M.Coult. (Cactaceae) in a rosetophyllus desert scrub in México. Journal of Arid Environments , 205 , 104824. https://doi.org/10.1016/j.jaridenv.2022.104824 Skouta, R., Morán-Santibañez, K., Valenzuela, C. A., Vasquez, A. H., & Fenelon, K. (2018). Assessing the antioxidant properties of larrea tridentata extract as a potential molecular therapy against oxidative stress. Molecules , 23 (7). https://doi.org/10.3390/molecules23071826 Sulistiyani, N. I., Ainurrofiq, A., & Suryanti, V. (2022). Antibacterial Activity of Chayote (Sechium edule Swartz) Squash Extracts and Their Phytochemical Constituents. Journal of Biodiversity and Biotechnology , 2 (1), 26. https://doi.org/10.20961/jbb.v2i1.61330 Torres-León, C., Rebolledo Ramírez, F., Aguirre-Joya, J. A., Ramírez-Moreno, A., Chávez-González, M. L., Aguillón-Gutierrez, D. R., Camacho-Guerra, L., Ramírez-Guzmán, N., Hernández Vélez, S., & Aguilar, C. N. (2023). Medicinal plants used by rural communities in the arid zone of Viesca and Parras Coahuila in northeast Mexico. Saudi Pharmaceutical Journal , 31 (1), 21–28. https://doi.org/10.1016/j.jsps.2022.11.003 Traversari, S., Cacini, S., Galieni, A., Nesi, B., Nicastro, N., & Pane, C. (2021). Precision Agriculture Digital Technologies for Sustainable Fungal Disease Management of Ornamental Plants. Sustainability , 13 (7), 3707. https://doi.org/10.3390/su13073707 Truong, D.-H., Nguyen, D. H., Ta, N. T. A., Bui, A. V., Do, T. H., & Nguyen, H. C. (2019). Evaluation of the Use of Different Solvents for Phytochemical Constituents, Antioxidants, and In Vitro Anti-Inflammatory Activities of Severinia buxifolia . Journal of Food Quality , 2019 , 1–9. https://doi.org/10.1155/2019/8178294 Tymczewska, A., Klebba, J., & Szydłowska-Czerniak, A. (2023). Antioxidant Capacity and Total Phenolic Content of Spice Extracts Obtained by Ultrasound-Assisted Extraction Using Deep Eutectic and Conventional Solvents. Applied Sciences , 13 (12), 6987. https://doi.org/10.3390/app13126987 Wang, Y., Su, Y., Shehzad, Q., Yu, L., Tian, A., Wang, S., Ma, L., Zheng, L., & Xu, L. (2023). Comparative study on quality characteristics of Bischofia polycarpa seed oil by different solvents: Lipid composition, phytochemicals, and antioxidant activity. Food Chemistry: X , 17 , 100588. https://doi.org/10.1016/j.fochx.2023.100588 Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-4370220\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":true,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":301044553,\"identity\":\"4b31384b-f1da-4dba-99ba-ff0fa8b47a4b\",\"order_by\":0,\"name\":\"Muyideen Olaitan Bamidele\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Universidad Autónoma de Coahuila\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Muyideen\",\"middleName\":\"Olaitan\",\"lastName\":\"Bamidele\",\"suffix\":\"\"},{\"id\":301044554,\"identity\":\"436d321c-561c-4c3e-be6f-55152f142258\",\"order_by\":1,\"name\":\"Olga B. Álvarez Pérez\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Universidad Autónoma de Coahuila\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Olga\",\"middleName\":\"B. Álvarez\",\"lastName\":\"Pérez\",\"suffix\":\"\"},{\"id\":301044555,\"identity\":\"e6042e28-5bb5-4117-b658-5d5457c12b2b\",\"order_by\":2,\"name\":\"José Sandoval-Cortes\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Universidad Autónoma de Coahuila\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"José\",\"middleName\":\"\",\"lastName\":\"Sandoval-Cortes\",\"suffix\":\"\"},{\"id\":301044556,\"identity\":\"a278ae02-6620-4aba-b485-9a9c4ad1c843\",\"order_by\":3,\"name\":\"María L. Flores-López\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Universidad Autónoma de Coahuila\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"María\",\"middleName\":\"L.\",\"lastName\":\"Flores-López\",\"suffix\":\"\"},{\"id\":301044557,\"identity\":\"8e70b3ea-e807-4bd3-b022-2924910c907c\",\"order_by\":4,\"name\":\"Mónica L. Chavez-González\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Universidad Autónoma de Coahuila\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Mónica\",\"middleName\":\"L.\",\"lastName\":\"Chavez-González\",\"suffix\":\"\"},{\"id\":301044558,\"identity\":\"c887f650-0c94-488f-8378-f64041ade24b\",\"order_by\":5,\"name\":\"Cristóbal N. Aguilar\",\"email\":\"data:image/png;base64,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\",\"orcid\":\"\",\"institution\":\"Universidad Autónoma de Coahuila\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Cristóbal\",\"middleName\":\"N.\",\"lastName\":\"Aguilar\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2024-05-05 04:09:06\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-4370220/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-4370220/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":56352716,\"identity\":\"829e9edd-2f0a-4d6b-b58a-732754012b23\",\"added_by\":\"auto\",\"created_at\":\"2024-05-13 04:59:29\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":13005,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eHydrolysable tannin composition in extracts obtained from macerated \\u003cstrong\\u003e(\\u003c/strong\\u003eI) and Soxhlet (II) extracted leaves and stems of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e. Soxhlet Ethyl acetate extract of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e stems (SOX ELS), Soxhlet Ethyl acetate extract of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e leaves (SOX ELL), Soxhlet \\u003cem\\u003en\\u003c/em\\u003e-hexane extract of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e leaves (Sox HLL), Soxhlet \\u003cem\\u003en\\u003c/em\\u003e-hexane extract \\u003cem\\u003eL. tridentata\\u003c/em\\u003e stems (Sox HLS), Soxhlet Diethyl ether extract of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e leaves (Sox DLL) and Soxhlet Diethyl ether extract of\\u003cem\\u003e L. tridentata\\u003c/em\\u003e stems (Sox DLS).\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4370220/v1/714281b0447a577c972a074f.png\"},{\"id\":56352720,\"identity\":\"ab389ce8-8d71-48cb-9053-c59fa4046d22\",\"added_by\":\"auto\",\"created_at\":\"2024-05-13 04:59:31\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":35760,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eThe condensed tannins composition of the macerated \\u003cstrong\\u003e(\\u003c/strong\\u003eI) and soxhlet \\u003cstrong\\u003e(\\u003c/strong\\u003eII) extracted leaves and stems extracts. Soxhlet Ethyl acetate \\u003cem\\u003eL. tridentata\\u003c/em\\u003e stems (ELS), Soxhlet Ethyl acetate \\u003cem\\u003eL. tridentata\\u003c/em\\u003e leaves (ELS), \\u003cem\\u003en\\u003c/em\\u003e-hexane\\u003cem\\u003e L. tridentata\\u003c/em\\u003e leaves (HLL), \\u003cem\\u003en\\u003c/em\\u003e-hexane\\u003cem\\u003e L. tridentata\\u003c/em\\u003e stems (HLS), Diethyl ether \\u003cem\\u003eL. tridentata\\u003c/em\\u003e leaves (DLL) and Diethyl ether \\u003cem\\u003eL. tridentata\\u003c/em\\u003e stems (DLS)\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4370220/v1/4e6739e3891568c1886b268a.png\"},{\"id\":56352719,\"identity\":\"78a36182-064d-4b1e-a3f9-8dbf8fd42946\",\"added_by\":\"auto\",\"created_at\":\"2024-05-13 04:59:31\",\"extension\":\"png\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":274733,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eAntifungal activity of macerated and soxhlet leaves and stems extracts of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"3.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4370220/v1/e55205c83c557dbae6cf783f.png\"},{\"id\":56353140,\"identity\":\"816f493f-53e2-451a-a73f-7e47b658812d\",\"added_by\":\"auto\",\"created_at\":\"2024-05-13 05:15:34\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":1581525,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4370220/v1/776cdfea-7e2b-4111-a861-3848c371058a.pdf\"},{\"id\":56352717,\"identity\":\"491a4fe9-2c61-48ab-9001-aa69a9f37047\",\"added_by\":\"auto\",\"created_at\":\"2024-05-13 04:59:29\",\"extension\":\"png\",\"order_by\":1,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":102241,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"GraphicalAbstract.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4370220/v1/79f48ec492bfcd51dc1e74ac.png\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"\\u003cp\\u003ePolyphenolic Bioactive Compounds from Larrea tridentata (DC.) Coville: Extraction, Characterization, Antioxidant, and Antifungal Activities \\u003c/p\\u003e\",\"fulltext\":[{\"header\":\"Highlights\",\"content\":\"\\u003cul\\u003e\\n \\u003cli\\u003eThe antioxidant potential of the extract increased with the polarity of the solvent used in the extraction.\\u003c/li\\u003e\\n \\u003cli\\u003eThe polyphenolic phytochemicals, which are inherent in plants, have been implicated in antifungal activities.\\u003c/li\\u003e\\n \\u003cli\\u003eThe plant composition of hydrolysable and condensed tannins varies with plant species, locations, extraction solvents, extraction methods, and climatic conditions.\\u003c/li\\u003e\\n \\u003cli\\u003ePlant extracts exhibit potent fungistatic activity that inhibits the growth of polyphagic pathogenic fungi.\\u003c/li\\u003e\\n\\u003c/ul\\u003e\"},{\"header\":\"1. Introduction\",\"content\":\"\\u003cp\\u003ePlant fungal infections are a prevalent and economically significant problem in agriculture, horticulture, and natural ecosystems (Traversari et al., \\u003cspan citationid=\\\"CR56\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e). These infections can lead to a range of diseases with negative consequences for agricultural output, plant health, and the environment (Fisher et al., \\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e; Nazarov et al., \\u003cspan citationid=\\\"CR46\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e). Bioactive metabolites used in combating fungal diseases are regarded as safe, effective, and environmentally friendly (Devi et al., \\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003eBioactive compounds found in plants, microorganisms, and animals have attracted considerable attention because of their wide antimicrobial properties. These compounds include flavonoids, phenolic acids, tannins, carotenoids, sterols, alkaloids and terpenoids (Banwo et al., \\u003cspan citationid=\\\"CR6\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e; Echegaray et al., \\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e; Haruna \\u0026amp; Yahaya, \\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e). They are essential in biological processes, such as antioxidant, antibacterial, anticancer, and antidiabetic activities, as well as in pathogen defense (Almatroodi et al., \\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e; Bourais et al., \\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e). Plants produce these bioactive compounds as defense mechanisms in response to threats and stress (Aguirre-Becerra et al., \\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e; Niazian \\u0026amp; Sabbatini, \\u003cspan citationid=\\\"CR48\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e). Secondary metabolites such as phenolic compounds are derived from primary metabolites. These compounds are synthesized using precursors, such as amino acids, fatty acids, organic acids, nucleotides, and sugars. (Marchiosi et al., \\u003cspan citationid=\\\"CR41\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e; Ranner et al., \\u003cspan citationid=\\\"CR50\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e). Secondary metabolites undergo biogenetic processes such as shikimate, polyketide, and mevalonate pathways to form diverse phenolic structures (Fuloria et al., \\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e; Magray et al., \\u003cspan citationid=\\\"CR39\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e). These compounds are characterized by the presence of one or more hydroxyl (\\u0026ndash;OH) groups attached to a six-carbon aromatic ring (Dikpınar \\u0026amp; S\\u0026uuml;zge\\u0026ccedil;-Sel\\u0026ccedil;uk, \\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e; Gulcin, \\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e). Phenolic compounds are present either as unconjugated glycosides or bound to sugars, such as glucose, galactose, rhamnose, xylose, or arabinose in glycosidic forms (Gonz\\u0026aacute;lez-Sarr\\u0026iacute;as et al., \\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e). Phenolic acids, flavonoids, tannins, stilbenes, lignans, and coumarins are some examples of phenolic compounds (Mutha et al., \\u003cspan citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e). They are principally antioxidants because of their electron-donating phenolic groups, and are present in fruits, vegetables, herbs, and medicinal plants (Chiorcea‐Paquim et al., 2020). Polyphenolic compounds are known to provide numerous health benefits and can be extracted using various methods including maceration, Soxhlet extraction, maceration, ultrasound-assisted extraction, microwave-assisted extraction, and fermentation (Jha \\u0026amp; Sit, \\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e; Manousi et al., \\u003cspan citationid=\\\"CR40\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e). Each technique has specific advantages and limitations that impact the characteristics and effectiveness of the extracted compounds.\\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eLarrea tridentata\\u003c/em\\u003e is a perennial shrub that is native to the semi-desert regions of the United States and Mexico. It is also referred to as the creosote bush, and its extensive medicinal usage dates back through the histories of both nations (Lasch\\u0026eacute; et al., \\u003cspan citationid=\\\"CR34\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e; Schwertner-Char\\u0026atilde;o et al., \\u003cspan citationid=\\\"CR52\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). This plant is part of the Zygophyllaceae family, which comprises approximately 30 genera and over 250 species that are predominantly discovered in warm and arid regions (Hadjadj et al., \\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e; Morales-Ubaldo, Rivero-Perez, et al., \\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). \\u003cem\\u003eL. tridentata\\u003c/em\\u003e typically grows at heights of 0.5 to 3.5 m and produces a light perfume(Bi\\u0026eacute; et al., \\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e). It has a spreading stem with lanceolate green-yellowish leaves, yellow blooms, and ovoid fruits with tiny white hairs enclosing black seeds(Skouta et al., \\u003cspan citationid=\\\"CR53\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e). This plant has gained popularity because to its apparent usefulness in resolving a wide range of health problems(Kannenberg et al., \\u003cspan citationid=\\\"CR32\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). \\u003cem\\u003eL. tridentata\\u003c/em\\u003e has long been used to treat a variety of health issues, including infertility, rheumatism, arthritis, diabetes, gallstones, kidney stones, common colds, diarrhea, skin problems, obesity, pain, and inflammation. (Morales-Ubaldo, Gonzalez-Cortazar, et al., \\u003cspan citationid=\\\"CR42\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e; Torres-Le\\u0026oacute;n et al., \\u003cspan citationid=\\\"CR55\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eL. tridentata\\u003c/em\\u003e leaves have been found to have strong antifungal activity against various fungal strains, including \\u003cem\\u003eRhizoctonia solani\\u003c/em\\u003e. The lanolin extract showed complete inhibition of mycelia at 2000 mg/L, while cocoa butter and water extracts required 3000 and 8000 mg/L for the same inhibition(Castillo et al., \\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e). \\u003cem\\u003eL.\\u003c/em\\u003e tridentata leaves were also effective against \\u003cem\\u003ePythium spp\\u003c/em\\u003e., \\u003cem\\u003eColletotrichum truncatum\\u003c/em\\u003e, \\u003cem\\u003eC. coccodes\\u003c/em\\u003e, \\u003cem\\u003eAlternaria alternata\\u003c/em\\u003e, \\u003cem\\u003eFusarium verticillioides\\u003c/em\\u003e, \\u003cem\\u003eF. solani\\u003c/em\\u003e, \\u003cem\\u003eF. sambucinum\\u003c/em\\u003e, and \\u003cem\\u003eRhizoctonia solani\\u003c/em\\u003e (Osorio et al., \\u003cspan citationid=\\\"CR49\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e). According to Chavez-Soliz et al.(2014), aqueous leaf extracts of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e at concentrations of 1000 mg/L and 5000 mg/L notably reduced the severity of \\u003cem\\u003ePodosphaera xanthii\\u003c/em\\u003e, which causes powdery melon mildew. Galvan et al. (2014) reported that an aqueous extract of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e (at concentrations of 10% and 20%) showed significant antifungal effects against \\u003cem\\u003ePhytophthora capsici\\u003c/em\\u003e and \\u003cem\\u003eAspergillus flavus\\u003c/em\\u003e, achieving complete inhibition after 48, 72, and 96 h. Furthermore, the leaf extract of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e, either alone or in combination with potassium sorbate, effectively inhibited \\u003cem\\u003eA. flavus\\u003c/em\\u003e growth under pH conditions of 3, 4, and 5, exhibiting a synergistic action when combined. (Mungu\\u0026iacute;a et al., 2014). Francisco et al. (2015) assessed the \\u003cem\\u003ein vitro\\u003c/em\\u003e antifungal activity of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e water, ethanol, lanolin, and cocoa butter extracts against \\u003cem\\u003eP. cinnamomi\\u003c/em\\u003e, the ethanol extract completely inhibited mycelium, while the lanolin extract showed very low inhibition. \\u003cem\\u003eL. tridentata\\u003c/em\\u003e ethanol and dichloromethane extracts reduced fungal growth by 75\\u0026ndash;100% against \\u003cem\\u003eA. tenuissima\\u003c/em\\u003e, \\u003cem\\u003eA. niger\\u003c/em\\u003e, \\u003cem\\u003ePenicillium polonicum\\u003c/em\\u003e, and \\u003cem\\u003eRhizopus oryzae\\u003c/em\\u003e. Aguirre-Joya et al. (\\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e) used bioactive films comprising \\u003cem\\u003eL. tridentata\\u003c/em\\u003e polyphenols to achieve 50% inhibition of \\u003cem\\u003eA. alternata\\u003c/em\\u003e, \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e, \\u003cem\\u003eBotrytis cinerea\\u003c/em\\u003e, and \\u003cem\\u003eC. gloeosporioides\\u003c/em\\u003e.\\u003c/p\\u003e \\u003cp\\u003eThis study addressed the potential of bioactive compounds in plant defense and their biological applications by extracting antifungal polyphenolic bioactive compounds from \\u003cem\\u003eL. tridentata\\u003c/em\\u003e stems and leaves through maceration and Soxhlet extraction, assessing the total polyphenol content and antioxidant potential, and characterizing the extracts using high-performance liquid chromatography (HPLC).\\u003c/p\\u003e\"},{\"header\":\"2. Materials and Methods\",\"content\":\"\\u003cdiv id=\\\"Sec3\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.1 Chemicals and reagents\\u003c/h2\\u003e \\u003cp\\u003eAnalytical grade reagents, including anhydrous sodium carbonate (Na\\u003csub\\u003e2\\u003c/sub\\u003eCO\\u003csub\\u003e3\\u003c/sub\\u003e), sodium hydroxide (NaOH), sodium carbonate (Na\\u003csub\\u003e2\\u003c/sub\\u003eCO\\u003csub\\u003e3\\u003c/sub\\u003e), sodium nitrite (NaNO\\u003csub\\u003e2\\u003c/sub\\u003e), aluminum chloride (AlCl\\u003csub\\u003e3\\u003c/sub\\u003e), sodium hydroxide (NaOH), hydrochloric acid (HCl), and ammonia ferric sulphate ([FeNH\\u003csub\\u003e4\\u003c/sub\\u003e(SO\\u003csub\\u003e4\\u003c/sub\\u003e)\\u003csub\\u003e2\\u003c/sub\\u003eO.12 H\\u003csub\\u003e2\\u003c/sub\\u003eO]), were supplied by Sigma Aldrich (Toluca, M\\u0026eacute;xico). Other chemicals also purchased included ethanol, distilled water, potato dextrose agar, gallic acid, quercetin, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2\\u0026prime;-Azino-bis (3-ethylbenzthiazoline-6-sulphonic acid, (ABTS) 2,4,6-Tripyridyl-s-triazine (TPTZ), Tween 80, and Folin-Ciocalteu reagent.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec4\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.2 Plant materials\\u003c/h2\\u003e \\u003cp\\u003eThe plant material, encompassing leaves and stems, was procured in February 2023 at coordinates Latitude 25\\u003csup\\u003eo\\u003c/sup\\u003e10\\u0026prime;59.3\\u0026prime;\\u0026prime; and Longitude 102\\u003csup\\u003eo\\u003c/sup\\u003e45\\u0026prime;41.2\\u0026prime; in Parras de la Fuente, Coahuila, Mexico, by Dr. Cristian Torres, a researcher at the Research Center and Ethnobiological Garden (CIJE), Universidad Autonoma de Coahuila, Unidad Torre\\u0026oacute;n, Viesca, Coahuila.\\u003c/p\\u003e \\u003cp\\u003eThe plants were manual defoliation, the branches were washed with distilled water and air-dried in the laboratory for 14 d. Subsequently, the leaves and stems pulverised using an automatic grinder at the Department of Biotechnology, Faculty of Chemical Science, Autonomous University of Coahuila, Mexico. The resulting pulverized samples were stored in plastic bags in the dark at 4\\u0026deg;C until needed.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec5\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.3 Extraction of polyphenolic compounds\\u003c/h2\\u003e \\u003cdiv id=\\\"Sec6\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003e2.3.1 Maceration extraction\\u003c/h2\\u003e \\u003cp\\u003ePulverized samples of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e leaves and stems were extracted by maceration. 5 g of sample was dissolved in 100 mL of water at a solid-to-liquid ratio of 1:10 (w/v) in the shaker. The extraction was performed for 24 h at 30\\u0026deg;C with constant agitation at 200 rpm in a shaker. The process was repeated using mixture of water-to-ethanol ratios, namely 50:50, 40:60, and 0:100 (v/v) ethanol: water ratio.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec7\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003e2.3.2 Soxhlet extraction\\u003c/h2\\u003e \\u003cp\\u003eThe extraction equipment included a distillation round flask, a glass condensed linked to 4\\u0026deg;C water, a thimble for sample placement, and an electric heat system. The solid-to-liquid ratio is 1:5 (w/v). A total of 5 h was spent extracting 50 g of material in 250 mL of solvent. The solvents were selected in the order of their polarity differences: \\u003cem\\u003en\\u003c/em\\u003e-hexane, ethyl acetate and diethyl ether. After recovering the extracts, the solvent was removed under vacuum and kept at 20\\u0026deg;C for future analysis to prevent degradation.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec8\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.4 Total Polyphenolic Constituents of extracts\\u003c/h2\\u003e \\u003cp\\u003eThe total polyphenolic components in each extract were determined using the technique described by Georg\\u0026eacute; et al. (2005) for hydrolysable and condensed tannins methodology proposed by Amaya-Chantaca et al. (2021). The total polyphenolic compounds per \\u003cem\\u003eL. tridentata\\u003c/em\\u003e (mg TPC/g \\u003cem\\u003eL. tridentata\\u003c/em\\u003e) were reported.\\u003c/p\\u003e \\u003cdiv id=\\\"Sec9\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003e2.4.1 Hydrolysable tannins (Folin Ciocalteu)\\u003c/h2\\u003e \\u003cp\\u003eThe total polyphenolic compounds were determined using the Folin Ciocalteu reagent, requiring only minor changes to the approach provided by Georg\\u0026eacute; et al. (2005). The Folin Ciocalteu test is an electron transfer technique that provides reducing capability, which is represented as phenolic content. To begin, 25 \\u0026micro;L of diluted sample, 25 \\u0026micro;L of Folin Ciocalteu reagent, and 25 \\u0026micro;L of sodium carbonate (0.7 M) were mixed. After 5 min, 125 \\u0026micro;L of distilled H\\u003csub\\u003e2\\u003c/sub\\u003eO was added and the sample was measured at 750 nm (UV-visible Epoch \\u003csup\\u003eTM\\u003c/sup\\u003e Microplate Spectrophotometer). According to linear regression using a calibration curve, the results were presented as milligrams of gallic acid equivalents per milli gram of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e (mg GAE/ g of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e).\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec10\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003e2.4.2 Condensed tannins (HCl-Butanol)\\u003c/h2\\u003e \\u003cp\\u003eCondensed tannins were quantified using the method described by Amaya-Chantaca et al. (2021) with a modification proposed by Swain and Hillis (1959). 250 \\u0026micro;L of water were combined with 1.5 mL of HCl: Butanol (1:9) and 1 mL of ferric reagent. The reaction mixture was heated to 95\\u0026deg;C for 45 min. The mixture is allowed to cool for 30 min before being read at 460 nm with a UV-visible spectrophotometer (UV-visible Epoch \\u003csup\\u003eTM\\u003c/sup\\u003e Microplate Spectrophotometer). According to linear regression using a calibration curve, the results were expressed as catechin g equivalents per gram (mg CE/g).\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec11\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.5 Antioxidant activity in the extracts\\u003c/h2\\u003e \\u003cdiv id=\\\"Sec12\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003e2.5.1 DPPH radical scavenging assay\\u003c/h2\\u003e \\u003cp\\u003eThe methodology was carried out with minor modifications according to the method proposed by Castro-Lopez et al. (2019); the electron donation capacity of the samples was evaluated from a purple colour solution of radical DPPH, using methanol as solvent (60 mM). Subsequently, 193 \\u0026micro;L of DPPH radical were added to each microplate for every 7 \\u0026micro;L of sample or standard curve (gallic acid). The reaction solution was subjected to an incubation period in the dark for 30 min; subsequently, the absorbance of the samples was recorded at a wavelength of 517 nm (UV\\u0026ndash;visible Epoch \\u0026trade; Microplate Spectrophotometer), and Trolox was used as a standard, and the results were expressed as mg Trolox equivalents per gram (mg TEAC /g)\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec13\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003e2.5.2 Ferric reducing ability (FRAP)\\u003c/h2\\u003e \\u003cp\\u003eThe ferric reducing ability was determined according to the method of Delgado-Andrade et al. (2005) with some modifications. The FRAP reagent was prepared by 2.5 mL of a 10 mM TPTZ solution in 40 mM HCl plus 2.5 mL of 20 mM FeCl\\u003csub\\u003e3\\u003c/sub\\u003e\\u0026sbquo; H\\u003csub\\u003e2\\u003c/sub\\u003eO and 2.5 mL of 0.3 M acetate buffer (pH 3.6). A 290 \\u0026micro;L of FRAP reagent was mixed with 10 \\u0026micro;L of the sample or standard concentration (Trolox). The reaction mix was incubated in darkness for 15 min and read by UV\\u0026ndash;visible Epoch \\u0026trade; Microplate Spectrophotometer automatic sample positioner (593 nm); the results were also expressed as milligrams equivalents of Trolox per gram of sample (mg TEAC/g).\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec14\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003e2.5.3 Radical Scavenging Activity ABTS\\u003c/h2\\u003e \\u003cp\\u003eA 7 mM ABTS solution was prepared, mixed with 2.45 mM K\\u003csub\\u003e2\\u003c/sub\\u003eS\\u003csub\\u003e2\\u003c/sub\\u003eO\\u003csub\\u003e8\\u003c/sub\\u003e, and left to incubate at room temperature for 12\\u0026ndash;16 h in the dark. The solution was diluted with ethanol until obtaining an absorbance of 0.7\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.02 at 734 nm. Then, 190 \\u0026micro;L of ABTS ethanolic solution and 10 \\u0026micro;L of the extracts were mixed, allowed to react for 1 min, and read at 734 nm in a microplate reader (Epoch, Biotek, Instruments, Inc.). The ABTS solution with the solvent of the samples was taken as a control. Trolox was used as a standard, and the results were expressed as mg Trolox equivalents per gram of extract (mg TEAC/g).\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec15\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.6 Antifungal assay of the extracts\\u003c/h2\\u003e \\u003cp\\u003eThe antifungal activities of the extracts were assayed against \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e and \\u003cem\\u003eA. alternata\\u003c/em\\u003e using the modified method of Eisa et al. (2017). Molten potato dextrose agar medium was poisoned with 0.5 mL of extracts mixed under aseptic conditions in laminar flow, and the medium was allowed to solidify. Each plate was inoculated with \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e and \\u003cem\\u003eA. alternata\\u003c/em\\u003e using a 5 mm thick disc of fungus (spores and mycelium) and incubated at 25\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1\\u0026deg;C until fungal growth in the control Petri plates was completed. The solvents used for the extraction were used as negative controls. The plates were incubated and monitored for 5\\u0026ndash;8 days to confirm the complete growth of the fungal. Each test was performed in triplicate by measuring the relative growth of the fungus in the treatment versus the control. The fungal mycelial growth (mm) in treated (T) and control (C) conditions was measured diametrically in three different directions, and the percentage inhibition in radial growth (I) was calculated using the Eq.\\u0026nbsp;1 (Vincent, 1947).\\u003c/p\\u003e \\u003cp\\u003e \\u003cspan class=\\\"InlineEquation\\\"\\u003e \\u003cspan class=\\\"mathinline\\\"\\u003e\\\\(I=\\\\frac{(C-T)}{C}*100\\\\)\\u003c/span\\u003e \\u003c/span\\u003eEq.\\u0026nbsp;1\\u003c/p\\u003e \\u003cp\\u003ewhere I\\u0026thinsp;=\\u0026thinsp;Inhibition of mycelia growth; C\\u0026thinsp;=\\u0026thinsp;Growth of fungus in control, and T\\u0026thinsp;=\\u0026thinsp;Growth of fungus in treatment.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec16\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.7 Reverse phase high-performance liquid chromatography RP-HPLC-ESI-MS analysis of extracts\\u003c/h2\\u003e \\u003cp\\u003eReverse phase high-performance liquid chromatography (HPLC) analyses were conducted using a Varian HPLC system, which included the following components: an autosampler (Varian ProStar 410, USA), a ternary pump (Varian ProStar 230I, USA), and a PDA detector (Varian ProStar 330, USA). Additionally, a liquid chromatograph ion trap mass spectrometer (Varian 500-MS IT Mass Spectrometer, USA) equipped with an electrospray ion source was used. The samples (5 \\u0026micro;L) were introduced into a Denali C18 column (150 mm \\u0026times; 2.1 mm, 3\\u0026micro;m, Grace, USA). The column temperature was maintained at 30\\u0026deg;C. Formic acid (0.2% v/v; solvent A) and acetonitrile (solvent B) were used as elution solvents. The following gradient program was applied: an initial composition of 3% B, linear increase to 9% B from 0 to 5 min, linear increase to 16% B from 5 to 15 min, and final linear increase to 50% B from 15 to 45 min. Subsequently, the columns were cleaned and conditioned. The flow rate was held constant at 0.2 mL/min, and detection was performed at wavelengths of 245, 280, 320, and 550 nm.\\u003c/p\\u003e \\u003cp\\u003eThe entire effluent, at a rate of 0.2 mL/min, was introduced into the mass spectrometer without splitting. All mass spectrometry (MS) experiments were performed in negative ionization mode ([M-H]\\u003csup\\u003e\\u0026minus;1\\u003c/sup\\u003e). Nitrogen served as the nebulizing gas and helium functioned as the damping gas. Specific ion source parameters included a spray voltage of 5.0 kV, capillary voltage of 90.0 V, and temperature of 350\\u0026deg;C. Data collection and processing were performed using MS Workstation software (Version 6.9).\\u003c/p\\u003e \\u003cp\\u003eSamples were initially analyzed in full scan mode across a mass-to-charge ratio (m/z) range of 50\\u0026ndash;2000. Phenolic compounds were identified by comparing the HPLC retention times with the MS data. Molecular weights were cross-referenced with literature and a database maintained by the Departamento de Investigaci\\u0026oacute;n en Alimentos at the Universidad Aut\\u0026oacute;noma de Coahuila (DIA-UAdeC).\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec17\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.8 Experimental design and statistical analysis\\u003c/h2\\u003e \\u003cp\\u003eThe experiments were carried out at three different measurement levels, and the outcomes are presented as the average value along with the standard deviation (SD). Data analysis involved the use of one-way analysis of variance (ANOVA), with the factor being the combination of cultivar and location. Statistical analysis was performed using the IBM SPSS Statistics software (SPPS Software).\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"3. Results and discussion\",\"content\":\"\\u003cdiv id=\\\"Sec19\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.1 Polyphenol composition (hydrolysable tannins)\\u003c/h2\\u003e \\u003cp\\u003eTannin may be found in the wood, stems, barks, leaves, roots, and fruits of any plant species, and its quantity varies by plant component. Tannin-rich plants are the most often utilized raw material for industrial-level tannin extraction (Zhang et al., 2023). Hydrolysable tannins offer a wide range of biological activities, including antioxidant, anti-inflammatory, and antifungal characteristics. This study revealed that the hydrolysable tannin content of macerated leaves of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e ranges from 2.72\\u0026ndash;6.41 mg GAE/g and the macerated stem extracts ranged from 0.93\\u0026ndash;1.64 mg GAE/g (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e(I)). The highest hydrolysable tannins were recorded with 60:40 ethanol: water 6.41\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.08 mg GAE/g (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e(I)). The macerated stem aqueous extract showed the lowest hydrolysable content. The macerated leaf extracts were 3.91 times more hydrolysable than the stems of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e. Plant leaves often contain a larger quantity of hydrolysable tannins than stems, roots, and flowers (Dettlaff et al., \\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e). Hydrolysable tannins are polyphenolic compounds that are present in various plant species. These tannins frequently act as plant defense mechanisms against herbivores and diseases, and they are more abundant in particular plant sections, with leaves being a typical reservoir for them (Savina et al., \\u003cspan citationid=\\\"CR51\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003eThe ethyl acetate leaf extract of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e contained the highest levels of hydrolysable tannins (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e(II)). This was followed by ethyl acetate and diethyl ether stem extracts. The extraction method and solvent polarity are among the factors that determine the composition of the extract (Akinmoladun et al., \\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e; Jasso de Rodr\\u0026iacute;guez et al., 2019). This result confirmed the effect of polarity on the composition of the extracts. The soxhlet ethyl acetate leaf extract (Sox ELL) contained 13.5 times hydrolysable tannins than the \\u003cem\\u003en\\u003c/em\\u003e-hexane leaf extracts (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e(II)). There is a possibility of bonding between the oxygen atom of ethyl acetate and hydrogen of the hydroxyl of the tannins leading to higher composition of tannins (Sulistiyani et al., \\u003cspan citationid=\\\"CR54\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). The SOX ELL contained the highest hydrolysable tannins 6.37\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.06 mg GAE/g of leaves \\u003cem\\u003eL. tridentata\\u003c/em\\u003e. This study obtained values of hydrolysable tannins higher than the work reported by Cadena et al. (\\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e). The phytochemical profile of a plant is influenced by genetics, environmental conditions, cultivation practices, harvesting time, post-harvest processing methods, plant parts, storage conditions, and biotic and abiotic stressors (Guan et al., \\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e; Liebelt et al., \\u003cspan citationid=\\\"CR36\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec20\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.2 Condensed tannins\\u003c/h2\\u003e \\u003cp\\u003eThe composition of condensed tannins in the macerated leaves and stems extracted from \\u003cem\\u003eL. tridentata\\u003c/em\\u003e is shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e. The values of condensed tannins vary from 0.75\\u0026ndash;3.29 CE/g in the leaves and 0.86\\u0026ndash;2.81 mg CE/ g in the stems (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e (I\\u0026amp;II). The leaves contained 1.17 times more condensed tannins in the leaves than stems. 60:40 ethanolic macerated leaves extracts gave the highest condensed tannins composition (3.30\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.012 mg CE/g). While also it equivalent in stems gave the best composition of condensed tannins (2.81 mg CE/g, Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e(I)). The plant composition of tannins varies on plant species and climatic circumstances(Bule et al., 2022). The aqueous extract of macerated stems contained more condensed tannins than hydrolysable tannins. Although, the concentration of condensed tannins were found throughout \\u003cem\\u003eL. tridentata\\u003c/em\\u003e in low concentrations(Hyder et al., \\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e2002\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003eThe condensed tannin content in soxhlet leaf extracts ranged from 0.96 to 3.32 mg CE/g, while the stems exhibited values between 0.69 and 2.90 mg CE/g (as shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e(II)). This indicates that the condensed tannin content in the leaves is higher than that in the macerated extracts. The use of polar organic solvents can enhance bonding with phytoconstituents, leading to an increased tannin content. The leaves had a condensed tannin content that was 1.14 times greater than that of the stems. Among the leaf extracts, the ethyl acetate extract contained the highest condensed tannin content compared to the \\u003cem\\u003en\\u003c/em\\u003e-hexane extract. This higher content in the ethyl acetate extract can be attributed to the polarity of the solvent used, as the hydroxyl hydrogen of condensed tannins can interact with the oxygen atom in ethyl acetate, thereby increasing tannin quantity. The condensed tannin content tends to increase as the polarity of the solvent used for extraction increases (Ng et al., \\u003cspan citationid=\\\"CR47\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e) .\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec21\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.3 Antioxidant Activity\\u003c/h2\\u003e \\u003cp\\u003ePlant extracts are rich in antioxidants including phenolic compounds, carotenoids, vitamins, and polyphenols. These compounds can scavenge free radicals, neutralize metal ions, and play a crucial role in preventing and controlling diseases caused by pathogens such as fungi, bacteria, and viruses. The antioxidant activity of plants is largely attributable to the presence of bioactive compounds. The antiradical action is primarily related to the polyphenolic content, and as the polarity of the solvent increases, the number of hydroxyl groups in the extract medium increases, as does the probability of hydrogen donation to free radicals(Albert et al., \\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e; Bautista-Hern\\u0026aacute;ndez et al., \\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e; Hodhodi et al., \\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). The DPPH assay is routinely used to evaluate the ability of antioxidants to scavenge free radicals. This involves quick electron transfer followed by delayed hydrogen transfer. The effectiveness of the hydrogen transfer process can be affected by the solvent used, particularly those capable of receiving hydrogen bonds such as ethanol or methanol (Isa et al., \\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e; Tymczewska et al., \\u003cspan citationid=\\\"CR58\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e). The extract with the highest scavenging potential was the macerated leaf extract of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e extracted with ethanol: water, 60:40 (191.48\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;3.37 mg TEAC/g), whereas the ethanolic stem extract showed the highest scavenging potential (166.29\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.69 mg TEAC/g) for the macerated stem extract. The macerated water extract showed the lowest antiradical activity in both leaves and stems of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e. SOX ELL extracts showed the highest scavenging activity (120.74\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;7.14 mg TEAC/g), whereas SOX HLS showed the lowest antioxidant activity (35.67\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.11 mg TEAC/g). The polarity of the solvent plays a crucial role in determining the antioxidant activity and extraction yield of the phytochemicals(Jeyaraj et al., \\u003cspan citationid=\\\"CR30\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e; Truong et al., \\u003cspan citationid=\\\"CR57\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e). Polar solvents tend to extract more polar components, which often possess stronger antioxidant properties, whereas nonpolar solvents tend to extract less polar components with weaker antioxidant activity(Lohvina et al., \\u003cspan citationid=\\\"CR38\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e; Wang et al., \\u003cspan citationid=\\\"CR59\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003eHowever, the leaves of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e possessed a higher antioxidant potential than the stems.\\u003c/p\\u003e \\u003cp\\u003eAntioxidants provided by \\u003cem\\u003eL. tridentata\\u003c/em\\u003e could serve as free radical scavengers and mitigate Reactive Oxygen Species /free radicals or could contribute to preventing the formation of hydroxyl radicals by deactivating free metal ions through chelation or converting H\\u003csub\\u003e2\\u003c/sub\\u003eO\\u003csub\\u003e2\\u003c/sub\\u003e to other innocuous compounds (such as water and oxygen).\\u003c/p\\u003e \\u003cp\\u003eThe ABTS radical-scavenging activity of the extracts showed a trend like that of DPPH. However, the value of the ABTS result of the antioxidant potential of the extracts was higher than that of the DPPH. This could be attributed to the incubation time (30 min) used in the assay. Skouta et al., (\\u003cspan citationid=\\\"CR53\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e) reported decreases in the DPPH scavenging ability with increase in incubation time. The 60:40 ethanol: water macerated leaf extract of \\u003cem\\u003eL. tridenatata\\u003c/em\\u003e (225.57\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;4.51 mg TEAC/g) showed the highest antioxidant activity compared to the water macerated leaf extract (106.07\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.97 mg TEAC/g). The macerated stem extract showed the highest antiradical activity at 50:50 ethanol: water (117.07\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;3.40 mg TEAC/g).\\u003c/p\\u003e \\u003cp\\u003eThe ELL extract showed the highest antiradical activity (226.57\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.04 mg TEAC/g) compared to the n-hexane soxhlet extract of leaf and stem, with antioxidant activity of (55.07\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.20 mg TEAC/g) and (43.57\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.26 mg TEAC/g) respectively. The FRAP test is a useful technique for determining the antioxidant capacities of diverse compounds. The reductive ability of the extract is consistent with that reported by (Skouta et al., \\u003cspan citationid=\\\"CR53\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e) with 60:40 ethanol: water having the highest reductive property (128.95\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.09 mg TEAC/g). The \\u003cem\\u003en\\u003c/em\\u003e-hexane extracts had fewer polyphenolic constituents and accounted for their low antioxidant potential. The polarity of the solvent determines the phytochemicals inherent in the extract and accounts for the higher antioxidant potential of the extract with higher solvent polarity.\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab1\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 1\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eThe antioxidant activities of extracts obtained from both maceration and Soxhlet extraction of leaves and stems of \\u003cem\\u003eLarrea tridentata\\u003c/em\\u003e. The values are reported as mean value with standard deviation.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"5\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eMethod Of Extraction And Plant Part\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eExtract\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eDPPH\\u003c/p\\u003e \\u003cp\\u003e(mg TEAC/g)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eABTS\\u003c/p\\u003e \\u003cp\\u003e(mg TEAC/g)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eFRAP\\u003c/p\\u003e \\u003cp\\u003e(mg TEAC/g)\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eMLL\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eAqueous\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e43.3\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;6.19\\u003csup\\u003ej\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e106.1\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.97\\u003csup\\u003ej\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e58.7\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.72\\u003csup\\u003eh\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e50% ET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e106.7\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.88\\u003csup\\u003ef\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e168.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.07\\u003csup\\u003ee\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e75.4\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.94\\u003csup\\u003ef\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e60% ET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e191.5\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;3.57\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e225.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;4.51\\u003csup\\u003eb\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e128.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.09\\u003csup\\u003ec\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e100% ET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e151.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.64\\u003csup\\u003ed\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e209.7\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2.84\\u003csup\\u003ec\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e87.5\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.79\\u003csup\\u003ee\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eMLS\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eAqueous\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e31.1\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2.94\\u003csup\\u003em\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e80.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2.02\\u003csup\\u003el\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e23.0\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.09\\u003csup\\u003el\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e50% ET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e106.7\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2.64\\u003csup\\u003ef\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e117.1\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;3.40\\u003csup\\u003eg\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e62.5\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.09\\u003csup\\u003eg\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e60% ET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e152.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.69\\u003csup\\u003ec\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e113.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;3.33\\u003csup\\u003eh\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e117.0\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.48\\u003csup\\u003ed\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e100% ET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e166.3\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.69\\u003csup\\u003eb\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e93.2\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2.02\\u003csup\\u003ek\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e15.0\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.94\\u003csup\\u003em\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"5\\\" rowspan=\\\"6\\\"\\u003e \\u003cp\\u003eSOXHLET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eELL\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e120.7\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;7.14\\u003csup\\u003ee\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e226.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.04\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e55.1\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.25\\u003csup\\u003ei\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eELS\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e103.3\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.11\\u003csup\\u003eg\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e195.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.01\\u003csup\\u003ed\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e21.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.63\\u003csup\\u003el\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eHLL\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e36.7\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;6.94\\u003csup\\u003ek\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e55.1\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.20\\u003csup\\u003em\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e49.3\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.48\\u003csup\\u003ej\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eHLS\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e35.7\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.11\\u003csup\\u003el\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e43.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.26\\u003csup\\u003en\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e34.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.48\\u003csup\\u003ek\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eDLL\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e85.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.69\\u003csup\\u003ei\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e154.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.50\\u003csup\\u003ef\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e150.5\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.25\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eDLS\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e88.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2.22\\u003csup\\u003eh\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e111.7\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;3.82\\u003csup\\u003ei\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e144.2\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.48\\u003csup\\u003eb\\u003c/sup\\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\\u003eMacerated L. tridentata leaves (MLL), macerated L. tridentata stems (MLS), Soxhlet Ethyl acetate \\u003cem\\u003eL. tridentata\\u003c/em\\u003e stems (ELS), Soxhlet Ethyl acetate \\u003cem\\u003eL. tridentata\\u003c/em\\u003e leaves (ELS), Soxhlet \\u003cem\\u003en\\u003c/em\\u003e-hexane \\u003cem\\u003eL. tridentata\\u003c/em\\u003e leaves (Sox HLL), Soxhlet \\u003cem\\u003en\\u003c/em\\u003e-hexane \\u003cem\\u003eL. tridentata\\u003c/em\\u003e stems (Sox HLS), Soxhlet Diethyl ether \\u003cem\\u003eL. tridentata\\u003c/em\\u003e leaves (Sox DLL) and Soxhlet Diethyl ether \\u003cem\\u003eL. tridentata\\u003c/em\\u003e leaves (Sox DLL).\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec22\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.4 Antifungal activities\\u003c/h2\\u003e \\u003cp\\u003ePlant extracts with high fungistatic activity inhibit the growth of polyphagic pathogenic fungi, demonstrating their potential as natural antifungal agents(Kursa et al., \\u003cspan citationid=\\\"CR33\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). Table\\u0026nbsp;\\u003cspan refid=\\\"Tab2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e, Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e: shows the effects of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e extract on the mycelial growth of \\u003cem\\u003eA. alternata\\u003c/em\\u003e and \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e. At 5% concentration, the SOX ELL extract inhibited \\u003cem\\u003eA. alternata\\u003c/em\\u003e by 79.12\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.014% and \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e by 80.71\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.38% (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e, Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e). The SOX ELS extract at 5% inhibited the mycelial growth of \\u003cem\\u003eA. alternata\\u003c/em\\u003e by 61.57\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.042% and \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e by 70.59\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.308%. Similarly, SOX DLL at 5% inhibited the mycelial growth of \\u003cem\\u003eA. alternata\\u003c/em\\u003e by 64.42\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.16% and \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e by 67.36\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.00%. The 5% SOX DLS extract inhibited \\u003cem\\u003eA. alternata\\u003c/em\\u003e by 57.49\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.27% and \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e by 65.60\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.17%. The 5%, 50:50 ethanol: water macerated leaf extract inhibited the mycelial growth of \\u003cem\\u003eA. alternata\\u003c/em\\u003e by 42.42\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.58% and \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e by 50.03\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.39%. In addition, 5%, 60:40 ethanol: water macerated leaf extract inhibited the mycelial growth of phytopathogens by 52.16\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.014% and 42.52\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.189%, respectively. However, the macerated stem extracts showed lower inhibition, ranging from 13\\u0026ndash;41% for \\u003cem\\u003eA. alternata\\u003c/em\\u003e and 0.04\\u0026ndash;15% for \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e.\\u003c/p\\u003e \\u003cp\\u003eThese results suggest that extracts with higher polyphenol content, such as SOX ELL, exhibited stronger fungicidal activity against \\u003cem\\u003eA. alternata\\u003c/em\\u003e and \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e. Additionally, the leaves generally showed higher inhibition than the stems, likely because of the higher polyphenol levels in the leaves. Conversely, aqueous extracts display low resistance to phytopathogens because of their low polyphenol content and limited ability to extract bioactive phytochemical compounds, possibly extracting more primary metabolites such as sugars. Liu et al., \\u003cspan citationid=\\\"CR37\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e reported low inhibition of aqueous extract on \\u003cem\\u003eAspergillus flavus\\u003c/em\\u003e. Guo et al., 2023, reported that fungicides with inhibition percentages between 75% and 90% are sensitive to phytopathogens and exhibit good fungicide efficiency.\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab2\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 2\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eThe Antifungal activities of macerated and Soxhlet extracts of Leaves and stems of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"4\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eMethod Of Extraction and Plant Part\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eExtracts\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e% Inhibition of\\u003c/p\\u003e \\u003cp\\u003e(\\u003cem\\u003eA. Alternata\\u003c/em\\u003e)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e% Inhibition of\\u003c/p\\u003e \\u003cp\\u003e(\\u003cem\\u003eF. Oxysporum\\u003c/em\\u003e)\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eMLL\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eAQUEOUS\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e19.5\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.31\\u003csup\\u003ei\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e11.2\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.52\\u003csup\\u003el\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e50% ET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e42.4\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.58\\u003csup\\u003ef\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e50.0\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.39\\u003csup\\u003ee\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e60% ET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e52.2\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.01\\u003csup\\u003ee\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e42.5\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.19\\u003csup\\u003ef\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e100% ET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e25.5\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.27\\u003csup\\u003eh\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e20.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.04\\u003csup\\u003ei\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eMLS\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eAQUEOUS\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e13.2\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.07\\u003csup\\u003ek\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.04\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.03\\u003csup\\u003en\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e50% ET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e41.1\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.07\\u003csup\\u003eg\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e12.8\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.28\\u003csup\\u003ek\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e60% ET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e25.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.03\\u003csup\\u003eh\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e9.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.26\\u003csup\\u003em\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e100% ET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e17.4\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.79\\u003csup\\u003ej\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e15.2\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.72\\u003csup\\u003ej\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"5\\\" rowspan=\\\"6\\\"\\u003e \\u003cp\\u003eSOXHLET\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eELL\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e79.1\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.01\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e80.7\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.38\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eELS\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e61.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.04\\u003csup\\u003ec\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e70.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.31\\u003csup\\u003eb\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eHLL\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e17.7\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.59\\u003csup\\u003ej\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e36.8\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.25\\u003csup\\u003eg\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eHLS\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.4\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.77\\u003csup\\u003el\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e24.20\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.59\\u003csup\\u003eh\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eDLL\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e64.4\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.16\\u003csup\\u003eb\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e67.36\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.00\\u003csup\\u003ec\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eDLS\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e57.5\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.27\\u003csup\\u003ed\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e65.60\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.17\\u003csup\\u003ed\\u003c/sup\\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 \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec23\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.5 Reverse phase high-performance liquid chromatography (HPLC)\\u003c/h2\\u003e \\u003cp\\u003eReverse-phase high-performance liquid chromatography (HPLC) of macerated and Soxhlet leaves and stems of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e revealed the presence of twenty-five (25) compounds (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e), while twenty-two compounds were recorded for the macerated and Soxhlet stem extracts (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e). The result revealed that most polyphenolic phytochemical and inherent in the leaves of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e. Which can be implicated for its antifungal effects. The class of compounds varies from lignans to catechins, acids, flavonols, and flavones. Fifteen (15) of these compounds were polyphenols, including caffeic acid 4-O-glucoside, rhamnetin, protocatechuic acid 4-O-glucoside, kaempferol, (+)-gallocatechin, luteolin, guteolin 7-O-(2-apiosyl-glucoside), gallic acid 4-O-glucoside-Coumaric acid 4-O-glucoside, quercetin, dihydroquercetin, piceatannol 3-O-glucoside, pterostilbene, tetramethylscutellarein, and cirsimaritin. Among the phenolic compounds of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e, a ligann such as NDGA stands out since it is extracted from the resin in leaves and stems and have been attributed to the biological activities of plants, particularly \\u003cem\\u003eL. tridentata\\u003c/em\\u003e (Lira et al., 2003). NDGA has been shown to inhibit several phytopathogenic fungi, and its potential mechanism of action has been explored. This phytochemical may decrease the growth and persistence of fungal colonies by interfering with the production of fungal biofilms. NDGA inhibits the formation of ergosterol, a critical component of fungal cell membranes. This disruption affects membrane integrity, resulting in decreased fungal growth and viability. The antifungal activity of NDGA is also attributed to its ability to inhibit microbial growth and cell function. Li et al. (\\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e) suggested that NDGA may inhibit the synthesis of pro-inflammatory molecules such as Tumor Necrosis Factor-alpha (TNF-α), which may contribute to its antifungal action.\\u003c/p\\u003e \\u003cp\\u003eFurthermore, NDGA's comprehensive strategy, which targets both membrane integrity and inflammatory pathways, makes it a good option for antifungal applications.\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab3\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 3\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eThe Reverse phase high-performance liquid chromatography (HPLC) analyses of macerated and soxhlet leaves extrcats of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e 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\\u003cp\\u003eGlycitein\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e283.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eMethoxyisoflavones\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eKaempferol\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e298\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eMethoxyflavonols\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eNordihydroguaiaretic acid (NDGA)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e301\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eLignans\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e(+)-Gallocatechin\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e305.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eCatechins\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e3-p-Coumaroylquinic acid\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e336.8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eHydroxycinnamic acids\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eLuteolin\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e285\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eFlavones\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eLuteolin 7-O-(2-apiosyl-glucoside)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e580.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eFlavones\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eMethylgalangin\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e284.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eMethoxyflavonols\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e3-p-Coumaroylquinic acid\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e336.8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eHydroxycinnamic acids\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eSecoisolariciresinol\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e364.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eLignans\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eGallic acid 4-O-glucoside\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e332.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eHydroxybenzoic acids\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eAvenanthramide 2c\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e314.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eHydroxycinnamic acids\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003ep-Coumaric acid 4-O-glucoside\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e325\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eHydroxycinnamic acids\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eDihydroquercetin\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e304\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eDihydroflavonols\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" 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\\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eMethylgalangin\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e284\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eMethoxyflavonols\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eGlycitein\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e283.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eMethoxyisoflavones\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eKaempferol\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e298\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eMethoxyflavonols\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eNDGA\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e301\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eLignans\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e(+)-Gallocatechin\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e305.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eCatechins\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e3-p-Coumaroylquinic acid\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e336.8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eHydroxycinnamic acids\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eLuteolin\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e285\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eFlavones\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eLuteolin 7-O-(2-apiosyl-glucoside)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e580.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eFlavones\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eMethylgalangin\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e284.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eMethoxyflavonols\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e3-p-Coumaroylquinic acid\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e336.8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eHydroxycinnamic acids\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eSecoisolariciresinol\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e364.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eLignans\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eGallic acid 4-O-glucoside\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e332.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eHydroxybenzoic acids\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eAvenanthramide 2c\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e314.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eHydroxycinnamic acids\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003ep-Coumaric acid 4-O-glucoside\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e325\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eHydroxycinnamic acids\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eDihydroquercetin\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e304\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eDihydroflavonols\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eQuercetin\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e300.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eFlavonols\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eTetramethylscutellarein\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e340.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eMethoxyflavones\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003eX\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eX - Present\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"4. CONCLUSION\",\"content\":\"\\u003cp\\u003eThe findings imply that \\u003cem\\u003eL. tridentata\\u003c/em\\u003e extracts have the potential to be employed in the control of plant diseases caused by \\u003cem\\u003eA. alternata\\u003c/em\\u003e and \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e. Notably, SOX ELL and SOX DLL extracts at 5% concentration revealed consistent effectiveness in lowering the frequency and severity of these plant diseases, consequently preventing their growth, and alleviating their negative effects on plants. To our knowledge, this is the first \\u003cem\\u003ein vitro\\u003c/em\\u003e evaluation of the polyphenol content, antioxidant potential, and antifungal activity of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e stems and leaves extracts. However, it is recommended that this study be expanded to the commercial greenhouse level by utilizing more ecologically friendly solvents.\\u003c/p\\u003e\"},{\"header\":\"Abbreviations\",\"content\":\"\\u003cp\\u003eHT-Hydrolysable tannins\\u003c/p\\u003e\\n\\u003cp\\u003eCT-condensed tannins\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eABTS- 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)\\u003c/p\\u003e\\n\\u003cp\\u003eFRAP- (Ferric Reducing Antioxidant Power)\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eDPPH - (2,2-diphenyl-1-picrylhydrazyl)\\u003c/p\\u003e\\n\\u003cp\\u003eHPLC - High-Performance Liquid Chromatography\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eRP-HPLC-ESI-MS - Reverse-Phase High-Performance Liquid Chromatography-Electrospray Ionization-Mass Spectrometry\\u003c/p\\u003e\\n\\u003cp\\u003eNDGA - Nordihydroguaiaretic acid\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eTPTZ - 2,4,6-Tris(2-pyridyl)-s-triazine,\\u003c/p\\u003e\\n\\u003cp\\u003eGAE/g - Gallic Acid Equivalents per gram\\u003c/p\\u003e\\n\\u003cp\\u003eSOX ELS - Soxhlet Ethyl acetate extract of L. tridentata stems\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eSOX ELL - Soxhlet Ethyl acetate extract of L. tridentata leaves\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eSox HLL - Soxhlet n-hexane extract of L. tridentata leaves\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eSox HLS - Soxhlet n-hexane extract of L. tridentata stems\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eSox DLL - Soxhlet Diethyl ether extract of L. tridentata leaves\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eSox DLS - Soxhlet Diethyl ether extract of L. tridentata stems\\u003c/p\\u003e\\n\\u003cp\\u003eCE/g - Condensed Tannin Equivalents per gram\\u003c/p\\u003e\\n\\u003cp\\u003eTEAC/g - Trolox Equivalent Antioxidant Capacity per gram\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eCRediT authorship contribution statement\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eMuyideen O. Bamidele: Conceptualization, Formal analysis, Investigation, Methodology, Writing – original draft. \\u0026nbsp;Olga B. Álvarez Pérez: Data curation, Resources, Visualization, Methodology, Supervision. José C. Sandoval: Data curation, Formal analysis, Methodology, Visualization, Supervision, Writing – review \\u0026amp; editing. María L. Flores-López: Formal analysis, Methodology, Supervision, Writing – original draft. Mónica L. Chavez-González: Methodology, Formal analysis,Supervision, Writing – review \\u0026amp; editing. Cristóbal N. Aguilar: Supervision, Data curation, Resources, Methodology, Formal analysis, Conceptualization, Writing – review \\u0026amp; editing, Writing – original draft.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAcknowledgements\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eMuyideen Olaitan Bamidele appreciated National Council of Humanities, Sciences and Technologies (CONACYT, Mexico) for PhD fellowship support awarded to me (1245491).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConflict of interest\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors declare no conflict of interest.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n \\u003cli\\u003eAguirre-Becerra, H., Vazquez-Hernandez, M. C., Saenz de la O, D., Alvarado-Mariana, A., Guevara-Gonzalez, R. G., Garcia-Trejo, J. F., \\u0026amp; Feregrino-Perez, A. A. 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Don (African Star Apple). \\u003cem\\u003eBulletin of the National Research Centre\\u003c/em\\u003e, \\u003cem\\u003e46\\u003c/em\\u003e(1), 40. https://doi.org/10.1186/s42269-022-00718-y\\u003c/li\\u003e\\n \\u003cli\\u003eAlbert, C., Codină, G. G., H\\u0026eacute;jja, M., Andr\\u0026aacute;s, C. D., Chetrariu, A., \\u0026amp; Dabija, A. (2022).\\u0026nbsp;Study of Antioxidant Activity of Garden Blackberries (Rubus fruticosus L.) Extracts Obtained with Different Extraction Solvents. \\u003cem\\u003eApplied Sciences\\u003c/em\\u003e, \\u003cem\\u003e12\\u003c/em\\u003e(8), 4004. https://doi.org/10.3390/app12084004\\u003c/li\\u003e\\n \\u003cli\\u003eAlmatroodi, S. A., Almatroudi, A., Alsahli, M. A., \\u0026amp; Rahmani, A. H. 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K., \\u0026amp; Ch\\u0026aacute;vez-Gonz\\u0026aacute;lez, M. L. (2022).\\u0026nbsp;Phenolic compounds and antioxidant activity of Lippia graveolens Kunth residual leaves fermented by two filamentous fungal strains in solid-state process. \\u003cem\\u003eFood and Bioproducts Processing\\u003c/em\\u003e, \\u003cem\\u003e136\\u003c/em\\u003e, 24\\u0026ndash;35. https://doi.org/10.1016/j.fbp.2022.09.001\\u003c/li\\u003e\\n \\u003cli\\u003eBi\\u0026eacute;, J., Sepodes, B., Fernandes, P. C. B., \\u0026amp; Ribeiro, M. H. L. (2023). Polyphenols in Health and Disease: Gut Microbiota, Bioaccessibility, and Bioavailability. \\u003cem\\u003eCompounds\\u003c/em\\u003e, \\u003cem\\u003e3\\u003c/em\\u003e(1), 40\\u0026ndash;72. https://doi.org/10.3390/compounds3010005\\u003c/li\\u003e\\n \\u003cli\\u003eBourais, I., Elmarrkechy, S., Taha, D., Mourabit, Y., Bouyahya, A., El Yadini, M., Machich, O., El Hajjaji, S., El Boury, H., Dakka, N., \\u0026amp; Iba, N. (2023). 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Comparative study on quality characteristics of Bischofia polycarpa seed oil by different solvents: Lipid composition, phytochemicals, and antioxidant activity. \\u003cem\\u003eFood Chemistry: X\\u003c/em\\u003e, \\u003cem\\u003e17\\u003c/em\\u003e, 100588. https://doi.org/10.1016/j.fochx.2023.100588\\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\":\"info@researchsquare.com\",\"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\":\"Hydrolysable tannins, Condensed tannins, Antifungal activity, Antioxidants \",\"lastPublishedDoi\":\"10.21203/rs.3.rs-4370220/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-4370220/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eThe significance of medicinal plants in inhibiting microbial growth in food and agricultural production as well as their economic viability cannot be overstated. These plants contain secondary metabolites, which are abundant in antimicrobial compounds, such as flavonoids, tannins, saponins, and alkaloids, and their extracts have demonstrated antimicrobial properties against a variety of plant pathogens.\\u003c/p\\u003e\\n\\u003cp\\u003eThe primary objective of this study was to explore the possibility of using bioactive compounds in plant defenses and their biological applications. To achieve this, antifungal polyphenolic bioactive compounds were extracted from the stems and leaves of \\u003cem\\u003eL. tridentata\\u003c/em\\u003e using conventional methods. The total polyphenol and antioxidant potential of the extracts were assessed and characterized using high-performance liquid chromatography (HPLC).\\u003c/p\\u003e\\n\\u003cp\\u003eThis study compared the polyphenolic constituents of extracts from emerging maceration and Soxhlet extraction techniques in the leaves and stems of \\u003cem\\u003eLarrea tridentata\\u003c/em\\u003e. The extracts were evaluated for total polyphenolic content (hydrolyzable (HT) and condensed tannins (CT)) and antioxidant activity (ABTS, FRAP, and DPPH). Reverse-Phase High-Performance Liquid Chromatography Electrospray Ionization coupled with mass spectrometry (RP-HPLC-ESI-MS) was used for qualitative identification of antimicrobial phytochemicals. Furthermore, the extracts were analyzed \\u003cem\\u003ein vitro\\u003c/em\\u003e for antifungal activity against \\u003cem\\u003eFusarium oxysporum\\u003c/em\\u003e and \\u003cem\\u003eAlternaria alternata\\u003c/em\\u003e.\\u003c/p\\u003e\\n\\u003cp\\u003eThe results revealed that 60:40 ethanol:water macerated leaf extract gave the highest hydrolysable tannins (6.41 ± 0.08 mg GAE/g), while its equivalent showed the highest condensed tannins (2.81 mg CE/g). Soxhlet ethyl acetate leaf (SOX ELL) extract showed 1.14 times more condensed tannin content than that of the stems. The antioxidant potential of the extract increased with increasing polarity of the extraction solvent. SOX ELL had higher antifungal effects against \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e and \\u003cem\\u003eA. alternata\\u003c/em\\u003e, whereas the 60:40 ethanol: water ratio resulted in 52% inhibition against \\u003cem\\u003eA. alternata\\u003c/em\\u003e and 43% inhibition against \\u003cem\\u003eF. oxysporum\\u003c/em\\u003e. Polyphenols with antifungal properties were found in the extracts, including caffeic acid 4-O-glucoside, rhamnetin, protocatechuic acid 4-O-glucoside, kaempferol, (+)-gallocatechin, luteolin, guteolin 7-O-(2-apiosyl-glucoside), gallic acid 4-O-glucoside, cumaric acid 4-O-glucoside, quercetin, NDGA, piceatannol 3-O-glucoside, pterostilbene, tetramethylscutellarein, and cirsimaritin.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cem\\u003eL. tridentata\\u003c/em\\u003e leaf extracts exhibit potential effectiveness in the development of biological control agents, which can not only enhance crop protection, but also contribute to overall agricultural sustainability.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Polyphenolic Bioactive Compounds from Larrea tridentata (DC.) Coville: Extraction, Characterization, Antioxidant, and Antifungal Activities\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2024-05-13 04:59:24\",\"doi\":\"10.21203/rs.3.rs-4370220/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"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\":\"500016a1-56f5-44e7-bb17-7328e237db7e\",\"owner\":[],\"postedDate\":\"May 13th, 2024\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"posted\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2024-05-13T04:59:27+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2024-05-13 04:59:24\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-4370220\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-4370220\",\"identity\":\"rs-4370220\",\"version\":[\"v1\"]},\"buildId\":\"qtupq5eGEP_6zYnWcrvyt\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}