Phytochemical Profiling and Bioactivity of Celtis caucasica: Antioxidant, Antidiabetic, Anticholinesterase, and Anti-inflammatory Potential

Phytochemical Profiling and Bioactivity of Celtis caucasica: Antioxidant, Antidiabetic, Anticholinesterase, and Anti-inflammatory Potential | 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 Article Phytochemical Profiling and Bioactivity of Celtis caucasica: Antioxidant, Antidiabetic, Anticholinesterase, and Anti-inflammatory Potential Sobia Gul, Atta Ur Rahman, Fahmeeda Kausar, Arshad Iqbal, Hina Gul, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6223687/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 This study evaluates the phytochemical composition and biological activities of the ethanol extract of Celtis caucasica leaves. Gas Chromatography-Mass Spectrometry (GC-MS) identified sixteen bioactive compounds in the ethanol extract of C. caucasica (ETCC), including 4-O-Methylmannose, Guaifenesin, Hexadecanoic acid derivatives, and Phytol, while the chloroform fraction of C. caucasica (CHFCC) contained twenty compounds, notably Phthalic acid di(2-propylpentyl) ester (11.94%) and Octadec-9-enoic acid (11.76%), known for their antioxidant, anti-inflammatory, and neuroprotective properties. Fourier-transform infrared (FT-IR) spectroscopy revealed diverse functional groups in ETCC and strong aromatic peaks with metal-organic complexes in CHFCC. Biological evaluations showed CHFCC had the highest acetylcholinesterase (AChE, 86.44%, IC₅₀ = 13.2 µg/mL) and butyrylcholinesterase (BChE, 92.67%, IC₅₀ = 9.66 µg/mL) inhibition, as well as potent α-glucosidase (88.61%, IC₅₀ = 11.99 µg/mL) and α-amylase inhibition (91.36%, IC₅₀ = 4.22 µg/mL), indicating strong antidiabetic potential. CHFCC also exhibited the highest COX-2 inhibition (88.61%), while the n-hexane fraction (NHFCC) showed the strongest 5-LOX inhibition (88.88%). Antioxidant assays revealed CHFCC had the highest radical scavenging activity (IC₅₀ = 77.24 µg/mL for DPPH and 60.67 µg/mL for ABTS), though lower than ascorbic acid (IC₅₀ = 2.81 µg/mL and 4.6 µg/mL, respectively). These findings highlight C. caucasica as a promising source of bioactive compounds with therapeutic potential. Biological sciences/Biochemistry Biological sciences/Chemical biology Biological sciences/Microbiology Biological sciences/Plant sciences Earth and environmental sciences/Biogeochemistry Celtus caucasica GC-MS Antioxidant Antidiabetic Anticholinesterase and Anti-inflammatory Properties Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Medicinal plants have long served as a valuable source for drug discovery, providing bioactive compounds with therapeutic potential 1 . In many developing countries, approximately 80% of primary healthcare relies on traditional medicines derived mainly from plant-based sources 2 . As global interest in natural products continues to rise, there is an increasing demand for medicinal plants. Diabetes mellitus has emerged as a major global health challenge, affecting both developed and developing nations. According to the World Health Organization (WHO), the number of diagnosed diabetes cases increased from 30 million in 1985 to 135 million in 1995, with projections estimating nearly 300 million cases by 2025 3 . This rising prevalence places a significant financial strain on healthcare systems, particularly in low-income regions 4 . Diabetes is classified into two main types: type I, which affects 5–10% of individuals and results from impaired insulin production, and type II, which accounts for approximately 90% of cases and arises from insulin resistance 5 . Persistent hyperglycemia, a hallmark of diabetes, can lead to severe complications, including damage to the central nervous system and blood vessels. Currently, several synthetic antidiabetic drugs, including meglitinides, biguanides, incretin mimetics, thiazolidinediones, sodium-glucose co-transporter 2 (SGLT2) inhibitors, and dipeptidyl peptidase-IV inhibitors, are under investigation for diabetes management 6 . While these medications have shown promising therapeutic effects, they are often associated with adverse side effects such as headaches, diarrhea, hypoglycemia, impotence, and vision impairment 7 . Given these challenges, there is a growing interest in developing safer, more effective antidiabetic treatments derived from natural sources. Traditional medicines, particularly herbal plants and their extracts have been recognized as viable alternatives to synthetic drugs due to their safety and efficacy. Notably, approximately 90% of modern medications are directly or indirectly derived from plant-based compounds, reinforcing their potential for new drug development 8 . Continued research on plant-based antidiabetic compounds is essential to discovering novel therapeutic agents with minimal side effects and enhanced clinical benefits 9 . Alzheimer’s disease (AD) is a progressive neurodegenerative disorder that significantly impairs the central nervous system 10 . Several factors contribute to AD pathogenesis, including oxidative stress, amyloid plaque accumulation, neuroinflammation, and cholinergic dysfunction 11 – 14 . While the exact cause of AD remains unclear, and no definitive cure has been identified, disease management focuses on addressing these underlying factors to slow its progression and alleviate symptoms 15 . Current treatments for AD do not provide a permanent cure but aim to mitigate cognitive decline, regulate behavioral symptoms, and improve overall quality of life 16 . Despite advancements in therapeutic strategies, further research is essential to develop more effective treatments. One key aspect of AD pathology is the disruption of cholinergic neurotransmission, which is crucial for normal cognitive function. Acetylcholine (ACh), a vital neurotransmitter, is degraded by the enzyme acetylcholinesterase (AChE), leading to impaired synaptic transmission and exacerbating AD symptoms 17 . AChE inhibitors have emerged as a promising therapeutic approach, as they help maintain acetylcholine levels in the brain, thereby improving cognitive function 18 . Numerous studies have highlighted the potential of plant-derived compounds in exhibiting anti-cholinesterase activity. Despite significant progress in pharmaceutical advancements, natural products continue to play a crucial role in drug discovery and healthcare. Increasing recognition of the therapeutic potential of medicinal plants has prompted pharmaceutical companies to explore plant-derived compounds as potential candidates for novel drug development 19 . Inflammation is a complex biological response that plays a critical role in the development and progression of various pathological conditions, including arthritis and cardiovascular diseases 20 . Current treatment strategies primarily rely on nonsteroidal anti-inflammatory drugs (NSAIDs), which inhibit cyclooxygenase (COX) enzymes. There are two COX isoforms: cyclooxygenase-1 (COX-1), which is constitutively expressed in most cells under normal physiological conditions, and cyclooxygenase-2 (COX-2), which is induced by pro-inflammatory mediators such as tumor necrosis factor-α (TNF-α), lipopolysaccharides (LPS), and tumor-promoting factors 21 . Efforts to develop alternative anti-inflammatory agents have led to the identification of both natural and synthetic dual COX-2/5-lipoxygenase (5-LOX) inhibitors, which exhibit promising anti-inflammatory activity. Research has focused on understanding the structural and pharmacological properties of natural compounds and heterocyclic systems that function as dual COX-2/5-LOX inhibitors 22 . However, a significant limitation of COX inhibition is its potential to stimulate 5-LOX, leading to increased leukotriene (LT) production. Although 5-LOX inhibitors have demonstrated protective effects, their widespread use is restricted due to various drawbacks 23 . The adverse effects associated with COX and 5-LOX inhibitors have thus limited their application in treating inflammatory disorders 24 . Given these challenges, there is an urgent need for new therapeutic strategies that effectively modulate inflammation while minimizing side effects. Celtis caucasica, commonly known as the Caucasian hackberry or Caucasian nettle tree, belongs to the Cannabaceae family. This medium-to-large deciduous tree is native to regions extending from Turkey to Central Asia and Assam, thriving in temperate climates 25 . Notable for its longevity and adaptability to rocky ecosystems and various environmental stressors, C. caucasica has been traditionally used in indigenous medicine. Its seeds and fruits have been employed to treat digestive ailments, including diarrhea and stomach disorders. Given its medicinal potential, further research into C. caucasica may uncover novel bioactive compounds with anti-inflammatory properties, contributing to the development of safer and more effective therapeutic alternatives. Materials and Methods Chemicals Instruments used The solvents employed in this study included ethanol, n-hexane, ethyl acetate, dichloromethane, chloroform (Master Chemical Supplier, Karachi, Pakistan), and distilled water (Islamia College Peshawar). Laboratory equipment utilized comprised glass funnels, filter papers, a thermostatically controlled water bath (STD/GMP), a digital magnetic stirrer (Model H3760-S), a vacuum rotary evaporator (Model RE-111, Labstac LLC, Pittsfield, MA, USA), and an analytical balance (Shimadzu, Karachi, Pakistan). GC-MS analysis was conducted using a Thermo GC Ultra (Version 5.0) system, coupled with a Thermo MS DSQ II and a GC 7890B-MS 5977B system. Plant Collection and Identification The fresh leaves of C. caucasica were collected in November from Islamia College Peshawar. The specimen was identified taxonomically by Dr. Naveed Akhtar, Associate Professor at the Department of Botany, Islamia College Peshawar. A voucher specimen (ICP/Bot-102) was deposited in the Department of Botany for reference. The leaves were then cut into small pieces and dried in the shade in a well-ventilated area. After drying, the leaves were ground into a coarse powder using a pestle and mortar to facilitate the maceration process. Approximately 1 kg of the powdered material was used for subsequent laboratory procedures 26 . Preparation of the crude extract One kilogram of powdered C. caucasica leaves was macerated in an appropriate volume of ethanol in a stainless-steel container, which was sealed and kept at room temperature for twelve days. To prevent solvent evaporation, the vessel was covered with a lid. The mixture was stirred three to four times a day using glass rods to ensure maximum extraction efficiency. After twelve days, the extract was first filtered through muslin cloth and then through Whatman No. 1 filter paper to remove plant residues. The filtrate was transferred to a clean container and concentrated under rotary vacuum at 50°C. The resulting extract was further dried in a water bath at 50°C, yielding 10 grams of dried crude ETCC. Fractionation Ten grams of crude ETCC was divided into two equal portions. One portion (5 g) was used as the crude methanolic extract, while the other portion underwent fractionation. The fractionation process began by dissolving the ETCC in distilled water. n-Hexane was then added to the mixture in a separating funnel, gently shaken, and left to settle for 10 minutes to separate the two phases: the n-hexane fraction and the aqueous layer. The n-hexane fraction was collected into a separate flask, and the remaining aqueous portion was further fractionated using a series of solvents, including dichloromethane, chloroform, n-hexane, and ethyl acetate, following the solvent-solvent fractionation method 27 . Each solvent-extracted fraction was concentrated using a rotary evaporator at appropriate temperatures. This process resulted in the isolation of the crude ETCC and its different solvent fractions: NHFCC, dichloromethane fraction (DCMFCC), ethyl acetate fraction (EAFCC), chloroform fraction (CHCC), and n-butanol fraction. In this study, we conducted phytochemical analysis of the ETCC, NHFCC, and CHCC fractions. GC-MS analysis The phytochemical composition of C. caucasica extracts, including the ETCC and CHFCC, was determined using GC-MS. The analysis was performed on a Thermo GC Ultra (Version 5.0) coupled with a Thermo MS DSQ II and a GC 7890B-MS 5977B system. A TR-5MS capillary column (30 m length, 0.25 µm film thickness, 0.25 mm internal diameter) was used for compound separation. High-purity helium gas (99.99%) served as the carrier at a flow rate of 1 mL/min. The injector was set to split mode at 250°C, and 1 µL of the sample was injected for analysis. The oven temperature program started at 50°C, held for 2 minutes, and then increased to 150°C at a rate of 8°C/min. The temperature was further raised to 300°C at 15°C/min and held for 5 minutes to ensure optimal separation of the compounds (Cupido et al., 2022). FTIR analysis FTIR analysis was conducted to identify the functional groups and structural changes in the molecules. Two grams of both the ETCC and CHFCC of C. caucasica were used for the analysis. To prepare the FTIR-grade KBr, 50 mg was mixed with 1 mg of the ETCC and CHFCC samples. The dried samples of each extract were then placed on the FTIR spectrometer, with a resolution of 4 cm − 1 and a scanning range of 4000–400 cm − 1 1 . α-amylase Inhibitory Activity Assay To assess the effect of C. caucasica extracts and column semi-purified fractions on α-amylase activity in an enzyme-starch system 28 , the following procedure was followed. A 1% concentration of each extract and semi-purified fraction of C. caucasica was mixed with 25 mL of a 4% potato starch solution in a beaker. Then, 100 mg of α-amylase was added to the starch solution and thoroughly mixed. The reaction mixture was incubated at 37°C for 60 minutes to allow enzymatic activity. After the incubation period, 0.1 M NaOH was added to stop the reaction. The mixture was then centrifuged at 3000 ×g for 15 minutes, and the supernatant was carefully decanted. Glucose concentration was subsequently measured to assess enzyme activity. α-glucosidase Inhibitory Activity Assay The α-glucosidase inhibitory activity of C. caucasica extracts and column semi-purified fractions was evaluated using the method described by Pistia-Brueggeman and Hollingsworth (2001), as reported by 29 . Inhibitory activity was assessed by incubating the extracts and fractions (62.5–1000 µg/mL) with a mixture containing 10 µL of α-glucosidase (1 U/mL) and 125 µL of phosphate buffer (0.1 M, pH 6.8) at 37°C for 20 minutes. The enzyme reaction was initiated by adding 20 µL of a 1 M p-nitrophenyl-β-D-glucopyranoside (pNPG) substrate solution, and the mixture was further incubated for 30 minutes under the same conditions. The reaction was halted by the addition of 50 µL of 0.1 N sodium carbonate (Na₂CO₃). Absorbance was measured at 405 nm using a UV-Vis spectrophotometer, both for the sample and control. The percentage of α-glucosidase inhibition was calculated using the formula: Inhibitory Activity (%) \(\:\frac{\text{O}\text{D}\left(\text{c}\text{o}\text{n}\text{t}\text{r}\text{o}\text{l}\right)-\text{O}\text{D}\left(\text{s}\text{a}\text{m}\text{p}\text{l}\text{e}\right)}{\text{O}\text{D}}\left(\text{c}\text{o}\text{n}\text{t}\text{r}\text{o}\text{l}\right))\times\:100\) Inhibitory Activity (%) \(\:\frac{\text{A}\text{b}\text{s}\text{C}-\text{A}\text{b}\text{s}\text{C}}{\text{A}\text{b}\text{s}\text{C}}\times\:100\) In-Vitro Anti-Cholinesterase Activity The inhibitory activities of AChE and BChE were assessed using a spectrophotometric method based on Ellman's assay. ETCC and its EAFCC and CHCC fractions were tested at concentrations of 62.5, 125, 250, 500, and 1000 µg/mL to evaluate their ability to inhibit AChE and BChE. The assay involves the enzymatic hydrolysis of acetylthiocholine iodide by the respective cholinesterase enzyme, resulting in the production of thiocholine. This liberated thiocholine reacts with Ellman's reagent, 5,5′-dithiobis-2-nitrobenzoic acid (DTNB), to form a chromogenic product that can be quantitatively measured spectrophotometrically. The end products of the reaction are 5-thio-2-nitrobenzoate and 2-nitrobenzoate-5-mercaptothiocholine. The absorbance of 5-thio-2-nitrobenzoate was measured at 412 nm using a spectrophotometer. Galantamine was used as a positive control, and plant extracts were assessed at the same concentrations. All samples were incubated at 37°C for 20 minutes before absorbance readings were taken. The change in absorbance over time was used to calculate enzyme inhibition 28 , 30 . The percentage of inhibition was calculated using the following formula: Enzyme inhibition (%) = 100 − percent enzyme activity Percent enzyme activity (%) = 100 × V/V max where (V max ) is an enzyme activity in the absence of an inhibitor. COX-2 Inhibitory Assay An in vitro assay was conducted to evaluate the anti-inflammatory activity of C. caucasica leaf extracts and column semi-purified fractions by assessing their inhibitory effect on cyclooxygenase-2 (COX-2) activity. The procedure outlined by 29 , was followed. A COX-2 enzyme solution was prepared at a concentration of 300 U/mL and chilled on ice for 5 to 10 minutes to activate the enzyme. Co-factors, including hematin (1 mM), N,N,N′,N′-tetramethyl-p-phenylenediamine (TMPD, 0.24 mM), and glutathione (0.9 mM), were dissolved in a Tris-HCl buffer (0.1 M, pH 8) and mixed with the activated enzyme solution. The plant extracts and fractions were added at concentrations ranging from 62.5 to 1000 µg/mL and incubated at 25°C for 5 minutes. To initiate the enzymatic reaction, 20 µL of a 30 mM arachidonic acid solution was introduced, and the mixture was incubated for an additional 4 to 5 minutes. Celecoxib was used as a positive control. The absorbance of the reaction mixture was measured at 540 nm using a UV-Vis spectrophotometer. Inhibitory Activity (%) \(\:\frac{\text{A}\text{b}\text{s}\text{C}-\text{A}\text{b}\text{s}\text{C}}{\text{A}\text{b}\text{s}\text{C}}\times\:100\) Here: AbsC = absorbance of control, while AbsS = absorbance of sample [extract/fractions]. LOX Inhibitory Assay In addition to the COX-2 assay, a 5-lipoxygenase (5-LOX) inhibitory assay was used to evaluate the in-vitro anti-inflammatory activity of C. caucasica extracts and semi-purified fractions, following the procedure described by 31 . Extracts and fractions were prepared at concentrations ranging from 62.5 to 1000 µg/mL. A 5-LOX enzyme solution was prepared at 10,000 U/mL, and the reaction was initiated by adding 80 mM linoleic acid as the substrate. The reaction mixture consisted of 250 µL of crude extract and 50 mM phosphate buffer (pH 6.3), to which 250 µL of enzyme solution was added. After 5 minutes of incubation, the substrate solution was introduced, and the mixture was well-mixed. Absorbance of both test and control samples was measured at 234 nm using a UV-Vis spectrophotometer. A concentration-response curve was constructed to determine the correlation between extract concentration and enzyme inhibition. The IC₅₀ value, representing the concentration inhibiting 50% of the enzyme activity, was calculated, and the percentage inhibition of 5-LOX activity was determined. Inhibitory Activity (%) \(\:\frac{\text{A}\text{b}\text{s}\text{C}-\text{A}\text{b}\text{s}\text{C}}{\text{A}\text{b}\text{s}\text{C}}\times\:100\) Here: AbsC = absorbance of control, while AbsS = absorbance of sample (extract/fractions). DDPH Free Radical Scavenging Assay The antioxidant activity of ETCC, NHFCC, and CHCC, and the standard compound was evaluated based on free radical scavenging activity against the stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, following the method described by 32 , 33 . The plant extracts and fractions were dissolved in ethanol, and ascorbic acid was used as a standard at concentrations ranging from 1 to 100 µg/mL. A 0.002% DPPH solution was prepared in methanol, and 1 mL of this solution was added to 1 mL of each plant extract/fraction and standard solution in separate test tubes. A blank control consisted of 1 mL of methanol and 1 mL of the 0.002% DPPH solution. The reaction mixtures were incubated at room temperature in the dark for 30 minutes, after which absorbance was measured at 517 nm using a UV-Vis spectrophotometer. ABTS Free Radical Scavenging Assay The antioxidant capacity of the compound was evaluated using the ABTS (2,2'-azinobis [3-ethylbenzthiazoline]-6-sulphonic acid) free radical assay, as described by 32 . In this method, antioxidants scavenge radical cations of ABTS, reducing absorbance at 734 nm. To prepare the ABTS solution, potassium persulfate (2.45 mM) and ABTS (7 mM) were mixed and incubated in the dark at room temperature for 12–16 hours to generate the radical cation solution. For the assay, the ABTS radical solution was diluted with phosphate buffer (0.01 M, pH 7.4) to achieve an absorbance of 0.70 at 734 nm. A mixture of 300 µL of the compound and 3.0 mL of the ABTS solution was added to a cuvette to measure the scavenging activity. The absorbance was recorded at 734 nm after 1 minute and again after 6 minutes of mixing. Ascorbic acid was used as a positive control. The assay was performed in triplicate, and the percentage of inhibition was calculated using the formula provided by 34 . Statistical analysis: Data are presented as the mean ± SEM, with significance determined using one-way ANOVA followed by Bartlett's test. Asterisks (*) denote statistically significant differences compared to the control group. A p-value of ≤ 0.05 was considered statistically significant. All samples were evaluated in triplicate. Results GC-MS analysis GC-MS analysis was conducted to characterize the phytochemical composition of the crude ETCC and its CHFCC. The mass spectra were compared with reference patterns from the National Institute of Standards and Technology (NIST) database, which includes over 62,000 compounds. Unidentified constituents (Figs. 1 and 2 ) were matched with known spectra from the NIST library to determine their molecular weight, structure, and chemical identity. Phytochemical composition of ETCC Phytochemical analysis of the ethanol extract of C. caucasica using GC-MS identified sixteen compounds belonging to various chemical classes. Many of these compounds are known for their significant bioactivities, including antioxidant, anti-inflammatory, anti-Alzheimer’s, and antimicrobial properties. The identified compounds include 4-O-Methylmannose (C₇H₁₄O₆) at RT 13.571, Thiophene, tetrahydro-2-methyl- (C₅H₁₀S) at RT 14.023, Benzoic acid, 2-ethylhexyl ester (C₁₅H₂₂O₂) at RT 14.602, Guaifenesin (C₁₀H₁₄O₄) at RT 15.243, Hexadecanoic acid, methyl ester (C₁₇H₃₄O₂) at RT 16.911, n-Hexadecanoic acid (C₁₆H₃₂O₂) at RT 17.264, Hexadecanoic acid, ethyl ester (C₁₈H₃₆O₂) at RT 17.577, 9,12-Octadecadienoic acid (Z,Z)-, methyl ester (C₁₉H₃₄O₂) at RT 18.458, Phytol (C₂₀H₄₀O) at RT 18.710, 12,15-Octadecatrienoic acid (Z,Z,Z)- (C₁₈H₃₀O₂) at RT 18.847, Oleic acid (C₁₈H₃₄O₂) at RT 18.881, Octadecanoic acid (C₁₈H₃₆O₂) at RT 19.115, Eicosanoic acid (C₂₀H₄₀O₂) at RT 20.825, and 1,9-Tetradecadiene (C₁₄H₂₆) at RT 22.148 (Fig. 1 , Table 1 ). Table 1 Phytochemical composition of ETCC RT %Area Name of Compound MF MW Peak Area Library 13.571 2.83 4-O-Methylmannose C 7 H 14 O 6 194.18 13627571 NIST20 14.023 14.57 Thiophene, tetrahydro-2-methyl- C 5 H 10 S 102.2 70158237 NIST20 14.602 2.09 Benzoic acid, 2-ethylhexyl ester C 15 G 22 O 2 234.33 10061527 NIST20 15.243 1.29 Guaifenesin C 10 H 14 O 4 198.22 6218972 NIST20 16.911 1.64 Hexadecanoic acid, methyl ester C 17 H 34 O 2 270.5 7878301 NIST20 17.264 6.61 n-Hexadecanoic acid C 16 H 32 O 2 256.42 31846853 NIST20 18710 3.83 Phytol C 20 H 40 O 296.5 18433814 NIST20 18.847 5.23 12,15-Octadecatrienoic acid, (Z,Z,Z)- C 18 H 30 O 2 278.4 25176956 NIST20 18.881 8.25 Oleic Acid C 18 H 34 O 2 82.5 39728445 NIST20 19.115 5.87 Octadecanoic acid C 18 H 36 O 2 284.5 28237421 NIST20 22.148 1.18 1,9-Tetradecadiene C 14 H 26 194.36 5674829 NIST20 24.558 2.79 Squalene C 30 H 50 410.7 13437612 NIST20 26.868 1.63 beta.-Tocopherol C 28 H 48 O 2 416.7 7857583 NIST20 28.338 14.09 Vitamin E C 6 H 8 O 6 176.12 67832529 NIST20 31.912 8.58 gamma.-Sitosterol C 29 H 50 O 414.7 41326518 NIST20 35.691 9.10 Benzoic acid, 2-(2,4-dihydroxybenzylidenamino)- C 14 H 11 NO 4 257.24 43807647 NIST20 Phytochemical composition of CHFCC Similarly, GC-MS analysis of the CHFCC confirmed the presence of approximately twenty compounds, as depicted in Fig. 2 and Table 2 . These compounds include Phthalic acid, di(2-propylpentyl) ester (11.94%), Octadec-9-enoic acid (11.76%), n-Hexadecanoic acid (6.38%), p-Xylene (5.29%), Ethylbenzene (4.90%), Benzene, nitro (2.81%), Nonacosane (6.20%), 9-Octadecenoic acid (Z)-, methyl ester (1.56%), and Benzoic acid, 2-ethylhexyl ester (1.07%), among others. Table 2 Phytochemical composition of CHFCC RT %Area Name of Compound MF MW Peak Area Library 2.356 1.62 Trichloromethane CHCl₃ 119.38 3096842 NIST20 2.551 4.90 Ethylbenzene C₈H₁₀ 106.17 9382483 NIST20 2.658 5.29 p-Xylene C 8 H 10 106.17 10128672 NIST20 2.725 1.30 Trichloromethane CHCl₃ 119.38 2485285 NIST20 2.959 1.98 p-Xylene C 8 H10 106.17 3781301 NIST20 5.837 2.81 Benzene, nitro C 6 H 5 NO 2 123.11 5372028 NIST20 12.356 0.80 Phenol,2,5-bis(1,1-dimethylethyl) C14H220 206.32 1526303 NIST20 12.525 0.85 Dodecanoic acid, methyl ester C₁₃H₂₆O₂ 214.35 1618672 NIST20 14.606 1.07 Benzoic acid, 2-ethylhexyl ester C₁₅H₂₂O₂ 234.34 2048960 NIST20 17.277 6.38 n-Hexadecanoic acid C₁₆H₃₂O₂ 256.43 12213770 NIST20 18.465 1.08 Methyl 10-trans,12-cis-octadecadienoate C₁₉H₃₆O₂ 296.49 2057046 NIST20 18.547 1.56 9-Octadecenoic acid (Z)-, methyl ester C₁₉H₃₆O₂ 296.49 2991241 NIST20 18.885 11.76 Octadec-9-enoic acid C₁₈H₃₄O₂ 282.47 22501628 NIST20 19.124 5.31 Octadecanoic acid C₁₈H₃₆O₂ 284.48 10152548 NIST20 22.105 11.94 Phthalic acid, di(2-propylpentyl) ester C₂₆H₄₂O₄ 410.6 22847404 NIST20 24.559 2.83 Squalene C₆₀H₁₀ 410.72 5412358 NIST20 25.503 6.20 Nonacosane C₂₉H₆₀ 404.88 11859419 NIST20 26.878 1.07 delta. -Tocopherol, O-methyl- C₂₉H₄₈O₃ 440.7 NIST20 28.349 18.43 Vitamin E C₂₉H₄₈O₂ 430.7 35257222 NIST20 31.909 6.82 beta. -Sitosterol C₂₈H₄₆O 414.68 13050602 NIST20 FT-IR analysis of ETCC The FTIR analysis of the C. caucasica ethanol extract identified several functional groups, including aromatic compounds, alcohols, amides, methylene groups, alkanes, ketones, and aldehydes. Notably, the majority of the peaks corresponded to alkane groups, as shown in Fig. 3 and Table 3 . Table 3 FTIR analysis of C. caucasica leaves ETCC. Peak Number Wavenumber (cm-1) Intensity Functional group Compound identifier 1 611.28341 62.91267 C–H bending aromatic compounds 2 879.65173 65.15021 C–H bending aromatic compounds 3 1028.74524 37.02025 C–O stretching alcohols, ethers, or esters 4 1088.38265 66.97643 C–O stretching alcohols, ethers, or esters. 5 1326.93227 87.81577 C–N stretching amide groups 6 1379.115 81.8381 C-H bending vibration methyl group 7 1416.38838 82.50645 C-H bending methyl or methylene group 8 1449.93442 82.78508 C–H Bending Alkanes 9 1654.938 92.89867 C = O stretching ketones, aldehydes, amides, or carboxylic acids 10 2832.77676 86.17968 H–C = O stretching aldehydes 11 2937.14222 82.16133 C–H Stretching methylene 12 2974.41559 75.88517 C–H stretching alkanes 13 3313.60334 68.73231 O–H stretching Alkanes FT-IR analysis of CHFCC The FTIR analysis of the CHFCC revealed characteristic absorption peaks, as detailed in Table 4 and Fig. 4 of the supplementary material. The wavelength values in the FTIR spectra correspond to specific chemical bonds within the molecule, which are identified through IR absorption. The chloroform extract of C. caucasica contains functional groups such as metal-organic complexes, alkanes, methylene groups, alcohol esters, and aromatic compounds. Notably, the most prominent peak was associated with aromatic compounds in the FTIR analysis. Table 4 FTIR analysis of C. caucasica leaves CHFCC. Peak Number Wavenumber (cm-1) Intensity Functional group Compound Identifier 1 469.64457 91.96578 metal-ligand stretching Not identified 2 667.19347 42.67843 C-H bending aromatic compounds 3 745.46757 3.47095 C–H bending aromatic compounds 4 928.10712 96.53538 C–H bending aromatic compounds 5 1028.74524 95.82001 C–O stretching alcohols, ethers, or esters 6 1215.11213 48.30565 C–O stretching esters, ethers, or alcohols 7 1423.84305 97.83264 C-H bending methyl group 8 1520.75384 98.59872 C–C stretching aromatic compounds 9 1602.75527 99.25807 C–C stretching aromatic compounds 10 2400.40557 99.83959 C–H stretching alkynes 11 3019.14365 95.66922 Metal-ligand vibrations metal-organic complexes or coordination compounds In-Vitro Anti-Cholinesterase Potential The ETCC, the n-hexane fraction (NHFCC), and the CHFCC demonstrated significant inhibitory effects against AChE and BChE at varying concentrations, as shown in Table 5 . The highest inhibition of AChE was observed at 500 µg/mL for CHFCC, with an inhibition of 86.44% and an IC50 value of 13.2. For BChE at the same concentration, CHFCC exhibited a 92.67% inhibition with an IC50 of 9.66. In comparison, ETCC showed 79.49% inhibition (IC50 16.5) against AChE and 82.61% inhibition (IC50 21.9) against BChE. NHFCC displayed 79.48% (IC50 27.4) inhibition of AChE and 79.37% (IC50 21.2) inhibition of BChE, respectively. Table 5 Percent inhibition of AChE and BChE by ETCC, NHFCC and CHFCC Solvents Concentration (µg/ml) AChE percent inhibition IC 50 (µg/ml) % BChE inhibition IC 50 (µg/ml) ETCC 500 79.49 16.5 82.61 21.9 250 75.58 77.60 125 72.93 72.83 62.5 65.44 57.55 31.25 54.56 54.58 NHFCC 500 79.48 27.4 79.37 21.2 250 75.62 74.72 125 70.60 70.47 62.5 57.68 65.50 31.25 51.72 48.46 CHFCC 500 86.44 13.2 92.67 9.66 250 81.08 88.58 125 79.84 82.54 62.5 74.94 78.20 31.25 52.58 73.40 Galantamine 500 94.40 6.8 93.08 3.06 250 85.03 86.45 125 80.90 80.58 62.5 76.44 75.40 31.25 71.22 70.80 The data are presented as the mean ± standard error of the mean (SEM). One-way ANOVA followed by Bartlett's test was used to assess significance. Asterisks denote the level of significance: "" (p < 0.001) indicates highly significant inhibition compared to the control, "" (p < 0.01) represents moderate significance, "" (p < 0.05) suggests mild significance, and "ns" (not significant) denotes no significant inhibition at certain concentrations. In-Vitro Anti-Diabetic Potential The anti-diabetic inhibitory activity of C. caucasica extracts against α-glucosidase and α-amylase was evaluated to assess their potential antidiabetic effects (Table 6 ). Among the extracts, CHFCC exhibited the highest inhibition of α-glucosidase (88.61% at 500 µg/mL) with an IC₅₀ of 11.99 µg/mL, followed by NHFCC (83.51%, IC₅₀ = 19.48 µg/mL) and ETCC (79.37%, IC₅₀ = 18.98 µg/mL). The standard drug acarbose showed the highest inhibition of α-glucosidase (94.08%) with an IC₅₀ of 11.98 µg/mL. In terms of α-amylase inhibition, CHFCC demonstrated the strongest activity (91.36% at 500 µg/mL) with an IC₅₀ of 4.22 µg/mL, followed by NHFCC (88.52%, IC₅₀ = 14.67 µg/mL) and ETCC (77.40%, IC₅₀ = 12.92 µg/mL). Acarbose, the control, showed the most potent inhibitory effect (97.23%) with an IC₅₀ of 9.72 µg/mL. Table 6 Percent inhibition of alpha glucosidase and alpha amylase by ETCC, NHFCC and CHFCC. Solvents Concentration (µg/mL) Alpha glucosidase percent inhibition IC 50 (µg/mL) Alpha amylase inhibition IC 50 (µg/mL) CHFCC 500 88.61 11.99 91.36 14.22 250 82.58 85.34 125 75.10 81.39 62.5 69.25 76.47 31.25 62.87 71.44 ETCC 500 79.37 18.98 77.40 12.92 250 73.37 72.57 125 69.30 67.36 62.5 63.42 62.56 31.25 53.52 57.37 NHFCC 500 83.51 19.48 88.52 14.67 250 75.76 82.4 125 67.22 74.54 62.5 63.51 67.34 31.25 56.37 61.30 Acarbose 500 94.08 11.98 97.23 9.72 250 87.45 92.45 125 81.58 85.90 62.5 76.40 81.00 31.25 71.80 76.90 In-Vitro Anti-Inflammatory Potential The anti-inflammatory activity of various C. caucasica extracts was assessed by evaluating their inhibitory effects on cyclooxygenase-2 (COX-2), cyclooxygenase-1 (COX-1), and 5-lipoxygenase (5-LOX) (Table 7 ). At a concentration of 500 µg/mL, CHFCC exhibited the highest inhibition of COX-2 (88.61%), followed by ETCC (79.08%) and NHFCC (75.32%). The standard drug, Celecoxib, demonstrated the highest inhibition of COX-2 (94.08%). Regarding COX-1 inhibition, CHFCC showed 69.58% inhibition, while NHFCC exhibited 61.30% inhibition. Celecoxib was found to be the most potent inhibitor of COX-1 among the samples. For 5-LOX inhibition, NHFCC demonstrated the highest activity (88.88%), followed by ETCC (79.37%) and CHFCC (73.80%). Montelukast, the positive control, showed the greatest 5-LOX inhibition (97.23%). Table 7 Percent inhibition of COX-2, COX-1 and 5-LOX by ETCC, NHFCC and CHFCC Solvents Concentration (µg/mL) COX-2 percent inhibition IC 50 COX-1 percent inhibition IC 50 5-LOX percent inhibition IC 50 (µg/mL) (µg/mL) (µg/mL) CHFCC 500 88.61 23.99 69.58 139.27 73.80 106.97 250 82.58 62.40 66.94 125 75.10 55.85 59.72 62.5 69.25 49.08 51.84 31.25 62.87 41.90 43.80 ETCC 500 79.08 43.76 54.93 673.93 79.37 60.68 250 72.56 45.94 72.37 125 65.03 37.90 65.30 62.5 61.90 31.51 58.42 31.25 53.42 22.80 50.52 NHFCC 500 75.32 77.25 61.30 286.38 88.88 32.84 250 67.12 55.78 83.54 125 62.79 49.44 75.01 62.5 55.79 42.72 67.68 31.25 47.20 34.29 59.82 Celecoxib 94.08 87.45 81.58 11.98 76.40 71.80 Montelukast 97.23 92.45 85.90 9.72 81.00 ± 0.30 76.90 ± 0.45 Indomethacin 91.36 20.91 85.34 78.39 72.47 65.44 DPPH and ABTS anti-radicals Assay The antioxidant capacity of various extracts was evaluated using the DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) assays to assess their radical scavenging activities (Table 8 ). Among all the samples tested, CHFCC exhibited the highest antioxidant activity, with IC₅₀ values of 77.24 µg/mL (DPPH) and 60.67 µg/mL (ABTS). ETCC and NHFCC showed moderate and lower antioxidant activities, respectively. Ascorbic acid, the positive control, demonstrated the highest antioxidant activity, with the lowest IC₅₀ values of 2.81 µg/mL for DPPH and 4.6 µg/mL for ABTS. Table 8 Antioxidant activity of ETCC, CHFCC and NHCS. Compound Concentration (µg/mL) DPPH percent inhibition IC 50 (µg/mL) ABTS percent inhibition IC 50 (µg/mL) CHFCC 500 75.32 ± 2.87 *** 77.24 79.36 ± 1.03 *** 60.67 250 67.12 ± 0.54 *** 72.36 ± 0.53 *** 125 62.78 ± 1.07 *** 65.31 ± 2.62 *** 62.5 55.78 ± 1.87 *** 58.41 ± 1.04 *** 31.25 47.21 ± 0.48 *** 50.53 ± 2.51 *** ETCC 500 60.34 ± 0.50 * 438.38 63.44 ± 0.58 * 250 51.26 ± 0.57 ns 55.48 ± 0.59 ns 328.35 125 43.40 ± 0.54 ns 46.22 ± 0.43 ns 62.5 34.39 ± 0.75 ns 37.49 ± 0.60 ns 31.25 27.23 ± 0.79 ns 31.47 ± 0.46 ns NHFCC 500 54.92 ± 1.72 * 673.92 57.39 ± 1.50 * 250 45.93 ± 0.90 ns 51.56 ± 3.83 ns 469.02 125 37.89 ± 2.31 ns 44.35 ± 0.54 ns 62.5 31.50 ± 0.58 ns 34.55 ± 0.94 ns 31.25 22.80 ± 1.41 ns 27.36 ± 1.09 ns Ascorbic acid 500 95.23 ± 0.89 *** 2.81 93.11 ± 0.54 *** 4.6 250 91.44 ± 0.42 ** 87.44 ± 0.42 *** 125 87.87 ± 0.81 ** 83.43 ± 0.39 ** 62.5 83.11 ± 0.13 * 78.89 ± 0.91 ** 31.25 77.87 ± 0.43 * 72.11 ± 0.19 * Data are presented as the mean ± SEM; one-way ANOVA followed by Bartlett's test was applied to determine significance; "***" indicates highly significant inhibition compared to control (p < 0.001), observed in CHFCC. "*" denotes moderate significance (p < 0.05), seen in ETCC and NHFCC. "ns" (not significant) suggests no statistically relevant inhibition at certain concentrations. Discussions Plant-based medicines have gained increasing popularity worldwide 35 , 36 , as researchers continually seek natural solutions that can be used to treat, diagnose, alleviate, or prevent various diseases 37 . Evaluating the therapeutic potential of these readily available natural compounds offers an opportunity to develop more affordable treatment options, especially in light of the current global economic challenges 38 . It is essential to recognize that enzyme inhibition in various biological compartments plays a crucial role in the pharmacodynamics of many drugs. In this study, fractions and extracts of C. caucasica were investigated for their phytochemical composition, as well as their antioxidant, anti-cholinesterase, anti-inflammatory, and anti-diabetic activities. In the in vitro anti-diabetic assay, different extracts and fractions of C. caucasica demonstrated significant (p < 0.05) dose-dependent inhibition of α-amylase and α-glucosidase activity when compared to the standard (Acarbose, IC₅₀: 11.98 and 9.72 µg/mL). The IC₅₀ values for the extracts ranged from 88.61–83.51 µg/mL for α-glucosidase and 91.36–88.52 µg/mL for α-amylase inhibition, respectively. These findings are consistent with previous research by 39 , which showed that Ficus benghalensis bark also had considerable inhibitory effects on α-glucosidase and α-amylase activity. Glucosidases are essential enzymes involved in various biological processes, such as the hydrolysis of dietary carbohydrates 40 . α-Glucosidase, found on the brush-border membrane of intestinal cells, catalyzes carbohydrate digestion 41 . The inhibitory effects of F. benghalensis bark on α-glucosidase have been attributed to its phenolic compounds and flavonoid content 42 . In this study, the fractions of C. caucasica were evaluated for their anti-inflammatory properties through two enzymatic assays, namely COX-2 and 5-LOX inhibition. The research focused on assessing the anti-inflammatory activities of various C. caucasica extracts based on their ability to inhibit COX-2, COX-1, and 5-LOX enzymes. The CHFCC extract exhibited the highest COX-2 inhibition at a concentration of 500 µg/mL, achieving 88.61% inhibition, which is comparable to the potency of the control drug, Celecoxib (94.08%). These results suggest that CHFCC contains potent COX-2 inhibitory compounds. Selective inhibition of COX-2 is crucial in anti-inflammatory therapy, as it helps avoid the adverse gastrointestinal effects often associated with COX-1 inhibition. Moderate COX-1 inhibition was observed with CHFCC (69.58%) and NHFCC (61.30%), indicating a selective inhibition pattern. These findings align with previous research, such as that of 43 , which reported a 25-fold selectivity for COX-2 over COX-1 in the ethanolic extract of Terminalia chebula fruit, with IC₅₀ values of 3.75 µg/mL for COX-2 and 90 µg/mL for COX-1. Regarding 5-LOX inhibition, NHFCC exhibited the highest activity (88.88%), suggesting the presence of bioactive compounds targeting the leukotriene pathway. This is consistent with studies on flavonoids, which are known to act as dual inhibitors of both COX-2 and 5-LOX, offering a comprehensive approach to inflammation treatment 44 . The dual inhibition of COX and 5-LOX pathways by C. caucasica extracts is particularly significant, as such compounds are likely to exhibit enhanced anti-inflammatory effects while minimizing side effects associated with selective enzyme inhibition. For example, curcumin has been shown to have dual inhibitory activity against both 5-LOX and COX-1/2 enzymes 45 . The diverse inhibitory activities of C. caucasica extracts against COX-2, COX-1, and 5-LOX highlight their potential as pharmaceutical anti-inflammatory agents. The strong inhibition of 5-LOX and excellent selectivity for COX-2 suggest that these extracts could be developed into safer and more effective anti-inflammatory therapies. AChE is an enzyme that plays a critical role in terminating nerve impulses in cholinergic synapses by hydrolyzing the neurotransmitter ACh into choline and acetic acid, particularly within the blood and neurological systems 46 . The findings from this study indicate that fractions of C. caucasica exhibit strong inhibitory activity against AChE and BChE, both of which are implicated in neurodegenerative diseases like Alzheimer's disease. Among the fractions tested, CHFCC was the most effective natural inhibitor of cholinesterase. Other fractions, including ETCC and NHFCC, also showed significant inhibitory activity, further supporting the therapeutic potential of C. caucasica. These results suggest that further investigation into the bioactive compounds in these fractions could lead to the development of natural anti-Alzheimer's drugs. Previous studies support the notion that plant-derived compounds can effectively inhibit cholinesterase enzymes. For instance, isoquinoline alkaloids like sanguinarine and chelerythrine, found in Macleaya cordata extracts, have demonstrated effective BChE inhibition, with IC₅₀ values ranging from 25.74 to 28.04 µg/mL 47 . Additionally, methanolic extracts of several Ayurvedic medicinal herbs have been shown to exhibit AChE inhibitory activity, highlighting the potential of plant-based inhibitors in treating cognitive disorders 48 . The pronounced inhibitory action of CHFCC against both AChE and BChE suggests that C. caucasica may contain bioactive compounds with dual inhibitory activity. Dual inhibition of these enzymes is especially beneficial in managing Alzheimer's disease, as both enzymes are involved in acetylcholine metabolism, and their concomitant inhibition could enhance cholinergic neurotransmission 49 . Moreover, in animal models, selective inhibition of BChE has been shown to elevate acetylcholine levels in the brain and improve cognitive function 50 . The antioxidant system of an organism plays a critical role in scavenging free radicals produced during normal metabolic processes. However, an excess of free radicals can lead to oxidative stress 51 . Antioxidants, which can be either enzymatic or non-enzymatic, neutralize the toxic effects of these free radicals. Plants possess a variety of antioxidant compounds that make up their defense system. In this study, the antioxidant potential of C. caucasica extracts (ETCC and CFCC) was assessed using the ABTS and DPPH scavenging assays. CHFCC demonstrated a high antioxidant potential, likely due to the presence of phenolic and flavonoid compounds, which are well known for their free radical scavenging abilities. Phenolic compounds exert their antioxidant activity by donating hydrogen atoms or electrons, stabilizing free radicals, and preventing damage caused by oxidative stress. This finding is consistent with studies on Cupressus species, which also exhibited substantial antioxidant capacity, particularly from ethyl acetate fractions, aligning with our observations for CHFCC 52 . ETCC exhibited moderate antioxidant activity, with IC₅₀ values of 438.38 µg/mL in the DPPH test and 328.35 µg/mL in the ABTS test. Literature suggests that the solvent used for extraction significantly impacts the yield and bioavailability of phenolic compounds 53 . While ETCC showed considerable antioxidant capacity, its lower efficiency may be attributed to differences in solvent-extractable polyphenolic content. NHFCC demonstrated the lowest antioxidant potential, with IC₅₀ values of 673.92 µg/mL (DPPH) and 469.02 µg/mL (ABTS), indicating a lower concentration of active antioxidant constituents. Similar trends have been observed in n-hexane extracts of medicinal plants, where non-polar fractions contain lower phenolic content and exhibit less radical scavenging activity 54 . To further confirm the presence of these phytochemicals, GC-MS was employed. The GC-MS analysis revealed 16 major compounds in the ethanolic extract and 20 in the chloroform extract. Notable bioactive compounds identified in ETCC include 4-O-methylmannose, which exhibits antimicrobial activity 55 , and octadecadienoic acid (Z,Z)-methyl ester, which has antimicrobial, antioxidant, and anti-inflammatory properties. Other compounds such as hexadecanoic acid, methyl ester, and n-hexadecanoic acid exhibit a range of bioactive properties, including antioxidant, nematicidal, pesticidal, and anti-inflammatory activities 56 , 57 . Additionally, phytol, identified in the extract, has antimicrobial activity and is known for its stability and low toxicity 58 , while glycerin, found in CHFCC, has antibacterial properties 59 . Other compounds like thiophene, tetrahydro-2-methyl, and squalene have been reported for their antimalarial, cytotoxic, and antioxidant activities 60 . Oleic acid, another identified compound, has various bioactive properties, including anti-inflammatory, antiandrogenic, and cancer-preventive effects 61 . Fourier-transform infrared (FTIR) spectroscopy is a valuable tool for identifying the types of chemical bonds and functional groups present in compounds. By analyzing the peak values in the infrared radiation region, the functional groups of active components can be identified, providing insights into the chemical composition of the sample. The FTIR spectrum of ETCC reveals a diverse array of phytochemicals, including flavonoids, terpenoids, and other polyphenolic compounds. These compounds are well-known for their antioxidant properties, which enable them to scavenge free radicals and alleviate oxidative stress 62 . Some flavonoids and terpenoids also exhibit antidiabetic properties, modulating carbohydrate metabolism and enhancing insulin sensitivity 62 . The presence of an amide group in the FTIR spectrum suggests the presence of alkaloids or peptides, which are associated with anticholinesterase activity and may be beneficial in treating neurodegenerative diseases 63 . Additionally, the prominence of aromatic compounds in CHFCC indicates a high concentration of phenolic constituents, known for their potent antioxidant activity. The antioxidant properties of phenolic compounds are primarily attributed to their hydroxyl groups, which donate hydrogen atoms to neutralize free radicals and mitigate oxidative stress 64 . Esters detected in the extract may contribute to its anti-inflammatory activity by modulating inflammatory processes. Some ester derivatives, such as those derived from ibuprofen, have been shown to possess enhanced anti-inflammatory properties compared to the parent compound, likely due to improved bioavailability and selective action 65 . Furthermore, the presence of metal-organic complexes suggests the potential existence of metalloproteins or coordination compounds. These complexes may play a role in various biochemical activities, such as enzymatic inhibition, which could contribute to anticholinesterase activity. Metalloproteins are involved in numerous physiological processes, and their interaction with metal ions is crucial for regulating enzymatic functions, including those involved in neurotransmission. Overall, FTIR analysis provides valuable insights into the chemical composition of ETCC, highlighting the presence of bioactive compounds that contribute to its antioxidant, anti-inflammatory, and potential anticholinesterase activities. These findings offer a deeper understanding of the therapeutic potential of C. caucasica in treating oxidative stress, inflammation, and neurodegenerative diseases. Conclusion The current research presents a thorough phytochemical and pharmacological analysis of C. caucasica leaf extracts, (ethanol extract, n-hexane fraction, and chloroform fraction). GC-MS analysis identified various bioactive compounds, including fatty acids, alcohols, and aromatic compounds, with known antioxidant, anti-inflammatory, and antimicrobial properties. The antioxidant activity, assessed via DPPH and ABTS assays, was highest in the chloroform fraction. The extracts also exhibited significant anti-cholinesterase and ant-diabetic potential, with CHFCC showing the strongest inhibition against α-glucosidase and α-amylase. In anti-inflammatory assays, CHFCC determined the highest inhibition of COX-2, while NHFCC showed the most potent 5-LOX inhibition. Future research should focus on in vivo validation of these effects, further investigation of molecular mechanism, formulation development for better bioavailability, toxicity and safety profiling, and exploring the synergistic effect of C. caucasica with other medicinal compounds. Declarations Ethics Approval and Consent to Participate Plant specimens of C. caucasica were obtained from Islamia College Peshawar, Pakistan. The samples were then transferred to the Department of Botany for taxonomic identification. Dr. Naveed Akhtar, Associate Professor, Department of Botany, Islamia College Peshawar, identified the plant species. A voucher specimen (ICP-BOT/102) was preserved in the departmental herbarium for future use. Author Contributions: Conceptualization, Methodology, and Data curation, S.G., A.U.R., F.K., A.I., and R.J.; Experiments, A.U.R., M.S.J., S.N., and S.T.; Visualization and Writing Original Draft, A.U.R, A.M.M.A., and R.J.; supervision and arranging resources, R.J., A.I., and K.M.K.; All authors have read and agreed to the published version of the manuscript. Acknowledgments: This work was carried out with the support of the “Cooperative Research Program for Agriculture Science and Technology Development (Project No. RS-2024-003222408)”, Rural Development Administration, Republic of Korea. The authors extend their appreciation to the Researcher Supporting Project number (RSPD2025R978), King Saud University, Riyadh, Saudi Arabia. Institutional Review Board Statement: Not applicable. Informed Consent Statement: Not Applicable. Data Availability: Data is provided within the manuscript. Conflict of Interest The author declared that there is no conflict of interest associated with this work. References Mittal, M., Gupta, N., Parashar, P., Mehra, V. & Khatri, M. Phytochemical evaluation and pharmacological activity of Syzygium aromaticum: a comprehensive review. Int. J. Pharm. Pharm. Sci. 6 , 67–72 (2014). Mujeeb, F., Bajpai, P. & Pathak, N. Phytochemical evaluation, antimicrobial activity, and determination of bioactive components from leaves of Aegle marmelos. BioMed research international 497606 (2014). (2014). Roglic, G. WHO Global report on diabetes: A summary. Int. J. Noncommunicable Dis. 1 , 3–8 (2016). Chew, N. W. et al. The global burden of metabolic disease: Data from 2000 to 2019. Cell Metabol. 35 , 414–428 (2023). Wang, J. et al. The protective effect of theaflavins on the kidney of mice with type II diabetes mellitus. Nutrients 15 , 201 (2022). Rath, P. et al. A critical review on role of available synthetic drugs and phytochemicals in insulin resistance treatment by targeting PTP1B. Appl. Biochem. Biotechnol. 194 , 4683–4701 (2022). Feingold, K. R. Oral and injectable (non-insulin) pharmacological agents for the treatment of type 2 diabetes. Endotext [Internet] (2024). Sharma, R. et al. Role of Shankhpushpi (Convolvulus pluricaulis) in neurological disorders: An umbrella review covering evidence from ethnopharmacology to clinical studies. Neurosci. Biobehavioral Reviews . 140 , 104795 (2022). DeFronzo, R. A. et al. Type 2 diabetes mellitus. Nat. reviews Disease primers . 1 , 1–22 (2015). Tran, N., Pham, B. & Le, L. Bioactive compounds in anti-diabetic plants: From herbal medicine to modern drug discovery. Biology 9 , 252 (2020). Sharma, R. et al. Traditional Ayurvedic and herbal remedies for Alzheimer’s disease: from bench to bedside. Expert Rev. Neurother. 19 , 359–374 (2019). Chen, Z. R., Huang, J. B., Yang, S. L. & Hong, F. F. Role of cholinergic signaling in Alzheimer’s disease. Molecules 27 , 1816 (2022). Qureshi, T. & Chinnathambi, S. Histone deacetylase-6 modulates Tau function in Alzheimer's disease. Biochim. et Biophys. Acta (BBA)-Molecular Cell. Res. 1869 , 119275 (2022). Sharma, R., Kabra, A., Rao, M. & Prajapati, P. Herbal and holistic solutions for neurodegenerative and depressive disorders: leads from Ayurveda. Curr. Pharm. Design . 24 , 2597–2608 (2018). Sharifi-Rad, J. et al. Multi-target mechanisms of phytochemicals in Alzheimer’s disease: Effects on oxidative stress, neuroinflammation and protein aggregation. J. Personalized Med. 12 , 1515 (2022). Kumar, A. & Singh, A. A review on Alzheimer's disease pathophysiology and its management: an update. Pharmacol. Rep. 67 , 195–203 (2015). Hassan, H. A. et al. Isolation and characterization of novel acetylcholinesterase inhibitors from Ficus benghalensis L. leaves. RSC Adv. 10 , 36920–36929 (2020). Taqui, R., Debnath, M., Ahmed, S. & Ghosh, A. Advances on plant extracts and phytocompounds with acetylcholinesterase inhibition activity for possible treatment of Alzheimer's disease. Phytomedicine plus . 2 , 100184 (2022). Valarezo, E. et al. Enantiomeric composition, antioxidant capacity and anticholinesterase activity of essential oil from leaves of Chirimoya (Annona cherimola Mill). Plants 11 , 367 (2022). Cacoub, P. et al. Towards a common definition for the diagnosis of iron deficiency in chronic inflammatory diseases. Nutrients 14 , 1039 (2022). Doiphode, S., Lokhande, K. B., Ghosh, P., Swamy, K. & Nagar, S. Dual inhibition of cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) by resveratrol derivatives in cancer therapy: in silico approach. J. Biomol. Struct. Dynamics . 41 , 8571–8586 (2023). Meshram, M. A., Bhise, U. O., Makhal, P. N. & Kaki, V. R. Synthetically-tailored and nature-derived dual COX-2/5-LOX inhibitors: Structural aspects and SAR. Eur. J. Med. Chem. 225 , 113804 (2021). El-Miligy, M. M. et al. Towards safer anti-inflammatory therapy: synthesis of new thymol–pyrazole hybrids as dual COX-2/5-LOX inhibitors. J. Enzyme Inhib. Med. Chem. 38 , 294–308 (2023). SL, M. Novel approach of multi-targeted thiazoles and thiazolidenes toward anti-inflammatory and anticancer therapy—dual inhibition of COX-2 and 5-LOX enzymes. Med. Chem. Res. 30 , 236–257 (2021). Cantarello, E. et al. Human impacts on forest biodiversity in protected walnut-fruit forests in Kyrgyzstan. J. Sustainable Forestry . 33 , 454–481 (2014). Ashafa, A., Sunmonu, T., Abass, A. & Ogbe, A. Laxative potential of the ethanolic leaf extract of Aloe vera (L.) Burm. f. in Wistar rats with loperamide-induced constipation. J. Nat. Pharmaceuticals . 2 , 158–162 (2011). Hartati, S., Angelina, M., Dewiyanti, I. & Meilawati, L. Isolation and characterization compounds from hexane and ethyl acetate fractions of Peperomia pellucida L. J. Trop. Life Sci. 5 , 117–122 (2015). Ellman, G. L., Courtney, K. D., Andres Jr, V. & Featherstone, R. M. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem. Pharmacol. 7 , 88–95 (1961). Jan, M. S. et al. Design, synthesis, in-vitro, in-vivo and in-silico studies of pyrrolidine-2, 5-dione derivatives as multitarget anti-inflammatory agents. Eur. J. Med. Chem. 186 , 111863 (2020). Badshah, S. L. et al. Isolation, characterization, and medicinal potential of polysaccharides of Morchella esculenta. Molecules 26 , 1459 (2021). Jan, M. S. et al. Synthesis of pyrrolidine-2, 5-dione based anti-inflammatory drug: in vitro COX-2, 5-LOX inhibition and in vivo anti-inflammatory studies. Latin Am. J. Pharm. 38 , 2287–2294 (2019). Souilah, N. et al. Phenolic compounds from an algerian endemic species of Hypochaeris laevigata var. hipponensis and investigation of antioxidant activities. Plants 9 , 514 (2020). Pena-Martín, C., Serrano, R., Grindlay, G. & Crespo, M. B. Assessment of antioxidant capacity of Cystosphaera jacquinotii (Fucales, Ochrophyta) by two methods: DPPH and CUPRAC. (2024). Shah, P. & Modi, H. Comparative study of DPPH, ABTS and FRAP assays for determination of antioxidant activity. Int. J. Res. Appl. Sci. Eng. Technol. 3 , 636–641 (2015). Dome, P., Tombor, L., Lazary, J., Gonda, X. & Rihmer, Z. Natural health products, dietary minerals and over-the-counter medications as add-on therapies to antidepressants in the treatment of major depressive disorder: a review. Brain Res. Bull. 146 , 51–78 (2019). Layek, B. & Mandal, S. Natural polysaccharides for controlled delivery of oral therapeutics: a recent update. Carbohydr. Polym. 230 , 115617 (2020). Fadilah, N. I. M., Isa, I. L. M., Zaman, W. S. W. K., Tabata, Y. & Fauzi, M. B. The effect of nanoparticle-incorporated natural-based biomaterials towards cells on activated pathways: A systematic review. Polymers 14 , 476 (2022). Iqbal, A. et al. Phytochemistry, Laxative, Prokinetic and Spasmolytic Verification of Withania somnifera (L.) Dunal Using In-Vitro, In-Vivo and In-Silico Approaches. J. Biol. Regul. Homeost. Agents . 38 , 3057–3072 (2024). Blickle, J., Andres, E. & Brogard, J. Current status of the treatment of type 2 diabetes mellitus. Alpha-glucosidase inhibitors. La. Revue de Med. Interne . 20 , 379s–383s (1999). Abadan, Ş. et al. Synthesis and molecular modeling studies of naphthazarin derivatives as novel selective inhibitors of α-glucosidase and α-amylase. J. Mol. Struct. 1278 , 134954 (2023). Dirir, A. M., Daou, M., Yousef, A. F. & Yousef, L. F. A review of alpha-glucosidase inhibitors from plants as potential candidates for the treatment of type-2 diabetes. Phytochem. Rev. 21 , 1049–1079 (2022). Daniel, R., Mathew, B., Devi, K. & Augusti, K. Antioxidant effect of two flavonoids from the bark of Ficus bengalensis Linn in hyperlipidemic rats. Indian J. Exp. Biol. 36 , 902–906 (1998). Mukhopadhyay, N., Shukla, A., Makhal, P. N. & Kaki, V. R. Natural product-driven dual COX-LOX inhibitors: Overview of recent studies on the development of novel anti-inflammatory agents. Heliyon 9 (2023). Md Idris, M. H. et al. Flavonoids as dual inhibitors of cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX): molecular docking and in vitro studies. Beni-Suef Univ. J. Basic. Appl. Sci. 11 , 117 (2022). Rudrapal, M. et al. Dual synergistic inhibition of COX and LOX by potential chemicals from Indian daily spices investigated through detailed computational studies. Sci. Rep. 13 , 8656 (2023). Mukherjee, P. K., Kumar, V., Mal, M. & Houghton, P. J. Acetylcholinesterase inhibitors from plants. Phytomedicine 14 , 289–300 (2007). Pohanka, M. Inhibitors of acetylcholinesterase and butyrylcholinesterase meet immunity. Int. J. Mol. Sci. 15 , 9809–9825 (2014). Mathew, M. & Subramanian, S. In vitro screening for anti-cholinesterase and antioxidant activity of methanolic extracts of ayurvedic medicinal plants used for cognitive disorders. PloS one . 9 , e86804 (2014). Tuzimski, T., Petruczynik, A., Szultka-Młyńska, M., Sugajski, M. & Buszewski, B. Isoquinoline alkaloid contents in Macleaya cordata extracts and their acetylcholinesterase and butyrylcholinesterase inhibition. Molecules 27 , 3606 (2022). Greig, N. H. et al. Selective butyrylcholinesterase inhibition elevates brain acetylcholine, augments learning and lowers Alzheimer β-amyloid peptide in rodent. Proceedings of the National Academy of Sciences 102, 17213–17218 (2005). Ayaz, M. et al. Anti-Alzheimer’s studies on β-sitosterol isolated from Polygonum hydropiper L. Front. Pharmacol. 8 , 697 (2017). Tawfeek, N. et al. Cupressus arizonica greene: phytochemical profile and cosmeceutical and dermatological properties of its leaf extracts. Molecules 28 , 1036 (2023). Anokwuru, C., Anyasor, G., Ajibaye, O., Fakoya, O. & Okebugwu, P. Effect of extraction solvents on phenolic, flavonoid and antioxidant activities of three nigerian medicinal plants. Nat. Sci. 9 , 53–61 (2011). Lagouri, V., Bantouna, A. & Stathopoulos, P. A comparison of the antioxidant activity and phenolic content of nonpolar and polar extracts obtained from four endemic lamiaceae species grown in Greece. J. Food Process. Preserv. 34 , 872–886 (2010). De Sousa, D. B. et al. Metabolomic Profile of Volatile Organic Compounds from Leaves of Cashew Clones by HS-SPME/GC-MS for the Identification of Candidates for Anthracnose Resistance Markers. J. Chem. Ecol. 49 , 87–102 (2023). Sudha, T., Chidambarampillai, S. & Mohan, V. GC-MS analysis of bioactive components of aerial parts of Fluggea leucopyrus Willd.(Euphorbiaceae). J. Appl. Pharm. Sci. 3 , 126–130 (2013). Chirumamilla, P., Dharavath, S. B. & Taduri, S. GC–MS profiling and antibacterial activity of Solanum khasianum leaf and root extracts. Bull. Natl. Res. Centre . 46 , 127 (2022). Ghaneian, M. T., Ehrampoush, M. H., Jebali, A., Hekmatimoghaddam, S. & Mahmoudi, M. Antimicrobial activity, toxicity and stability of phytol as a novel surface disinfectant. Environ. Health Eng. Manage. J. 2 , 13–16 (2015). Singh, B. R. in Proceedings of International conference and 28th Annual convention of IAVMI-2014 on Challenges and opportunities in animal health at the face of globalization and climate change, Department of Veterinary Microbiology and Immunology, DUVASU, Mathura, India. 26–29. Yasin, Z. N. M. et al. Biological Activities and GCMS Analysis of the Methanolic Extract of Christia vespertilionis (LF) Bakh. F. Leaves. Asian J. Med. Biomed. 4 , 78–88 (2020). Nallathambi, A. & Bhargavan, R. GC/MS analysis of bioactive compounds in aqueous extract of cynodon dactylon. Indian J. Public. Health Res. Dev. 10 , 55–59 (2019). Naveed, M. et al. Characterization and evaluation of the antioxidant, antidiabetic, anti-inflammatory, and cytotoxic activities of silver nanoparticles synthesized using Brachychiton populneus leaf extract. Processes 10 , 1521 (2022). Zlatić, N., Stanković, M. & Anticholinesterase Antidiabetic and anti-inflammatory activity of secondary metabolites of Teucrium species. Teucrium Species: Biology Appl. , 391–411 (2020). Zeb, A. & Zeb, A. Molecular Mechanism of Phenolic Antioxidants. Phenolic Antioxid. Foods: Chem. Biochem. Anal. , 413–434 (2021). Kravchenko, I., Kireva, M. & Alekseeva, E. Synthesis and anti-inflammatory activity of ibuprofen esters. Pharm. Chem. J. 48 , 313–316 (2014). Supplementary Material Supplementary Material are not available with this version. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6223687","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":433031363,"identity":"d9e706ba-5890-43ef-ae41-fa2f87810fef","order_by":0,"name":"Sobia Gul","email":"","orcid":"","institution":"Islamia College Peshawar","correspondingAuthor":false,"prefix":"","firstName":"Sobia","middleName":"","lastName":"Gul","suffix":""},{"id":433031364,"identity":"0b3ddfc7-948f-4f9c-ab74-42912c43e2d1","order_by":1,"name":"Atta Ur Rahman","email":"","orcid":"","institution":"Islamia College Peshawar","correspondingAuthor":false,"prefix":"","firstName":"Atta","middleName":"Ur","lastName":"Rahman","suffix":""},{"id":433031366,"identity":"d6450ef6-2e8e-44d6-87b0-301b42c3fbba","order_by":2,"name":"Fahmeeda Kausar","email":"","orcid":"","institution":"Islamia College Peshawar","correspondingAuthor":false,"prefix":"","firstName":"Fahmeeda","middleName":"","lastName":"Kausar","suffix":""},{"id":433031368,"identity":"3f29a37b-867f-4285-b18d-e6865631233b","order_by":3,"name":"Arshad Iqbal","email":"","orcid":"","institution":"Islamia College Peshawar","correspondingAuthor":false,"prefix":"","firstName":"Arshad","middleName":"","lastName":"Iqbal","suffix":""},{"id":433031369,"identity":"69318f41-2230-45c3-a627-7b8841c730e7","order_by":4,"name":"Hina Gul","email":"","orcid":"","institution":"Islamia College Peshawar","correspondingAuthor":false,"prefix":"","firstName":"Hina","middleName":"","lastName":"Gul","suffix":""},{"id":433031370,"identity":"f129d1ed-7976-492d-aa19-4cafe72c596f","order_by":5,"name":"Muhammad Saeed Jan","email":"","orcid":"","institution":"Bacha Khan University Charsadda","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"Saeed","lastName":"Jan","suffix":""},{"id":433031371,"identity":"e2af0fef-d169-41b8-8fe4-1b2985ddea30","order_by":6,"name":"Ashraf M. M. Abdelbacki","email":"","orcid":"","institution":"King Saud University","correspondingAuthor":false,"prefix":"","firstName":"Ashraf","middleName":"M. M.","lastName":"Abdelbacki","suffix":""},{"id":433031373,"identity":"b090290f-2dce-4e54-adb1-c41506149abc","order_by":7,"name":"Rahmatullah Jan","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5klEQVRIiWNgGAWjYFACHhBhw8PPzHwYKsJGjJaENDnJdrZkkrQcNjY4z2NMnBbz9rNHN/z8wZzYcJjns3FhG4M8fwNb2gd8WmTO5KXd7ElgS2xs5t2cPLONwXDGAbbDM/BpkWDIMbvBk8CT2MzMu/kwbxsD4wYG9ma8DpPgf2N280+CRGIbM89jkBZ7wlokcsxu8yQYGPMw8zAnA7UkbmBgO0xAyxuz2zJpCXISzGzGxjznJJJnHIaHNi6H5ZjdfGPzn8f+/OHH0jxlNrb97W3GeLVgGMHAwEyShlEwCkbBKBgF2AAAxa8++44SXhEAAAAASUVORK5CYII=","orcid":"","institution":"kyungpook National University","correspondingAuthor":true,"prefix":"","firstName":"Rahmatullah","middleName":"","lastName":"Jan","suffix":""},{"id":433031374,"identity":"01eccd09-e75b-4e4b-b690-81a9e95d0634","order_by":8,"name":"Kyung-Min Kim","email":"","orcid":"","institution":"kyungpook National University","correspondingAuthor":false,"prefix":"","firstName":"Kyung-Min","middleName":"","lastName":"Kim","suffix":""}],"badges":[],"createdAt":"2025-03-14 05:08:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6223687/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6223687/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":79461477,"identity":"4216bfdb-b6d1-47bc-a157-142c2cde7f88","added_by":"auto","created_at":"2025-03-28 17:24:29","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":154109,"visible":true,"origin":"","legend":"\u003cp\u003eGas Chromatogram of ETCC.The numbers represent the retention times of individual compound.\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-6223687/v1/aa7b53c3f7bb954222194eaa.png"},{"id":79461471,"identity":"b42736c0-fa0a-40f5-8f41-b6fbd381700d","added_by":"auto","created_at":"2025-03-28 17:24:29","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":141100,"visible":true,"origin":"","legend":"\u003cp\u003eGas Chromatogram of CHFCC. The numbers represent the retention times of individual compound.\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-6223687/v1/9f4936d57397bb78e623690d.png"},{"id":79461475,"identity":"e00ddc7d-dc18-4b51-993f-c078a5179b1d","added_by":"auto","created_at":"2025-03-28 17:24:29","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":206194,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR spectrum of \u003cem\u003eC. caucasica\u003c/em\u003e ETCC\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-6223687/v1/a75f70f9f7953edca20a7eb9.png"},{"id":79461811,"identity":"b3e4abec-723f-409e-ba9b-3ffbba875b70","added_by":"auto","created_at":"2025-03-28 17:32:29","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":164825,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR spectrum of \u003cem\u003eC. caucasica\u003c/em\u003e CHFCC\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-6223687/v1/ae30565cd6cf14d97841e562.png"},{"id":83761403,"identity":"9c9f33ef-342f-4ec5-b7c5-93daa65831b0","added_by":"auto","created_at":"2025-06-02 09:38:53","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2275109,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6223687/v1/9ef4e386-dd77-40c2-abec-801fb1a70032.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Phytochemical Profiling and Bioactivity of Celtis caucasica: Antioxidant, Antidiabetic, Anticholinesterase, and Anti-inflammatory Potential","fulltext":[{"header":"Introduction","content":"\u003cp\u003eMedicinal plants have long served as a valuable source for drug discovery, providing bioactive compounds with therapeutic potential \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. In many developing countries, approximately 80% of primary healthcare relies on traditional medicines derived mainly from plant-based sources \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. As global interest in natural products continues to rise, there is an increasing demand for medicinal plants. Diabetes mellitus has emerged as a major global health challenge, affecting both developed and developing nations. According to the World Health Organization (WHO), the number of diagnosed diabetes cases increased from 30\u0026nbsp;million in 1985 to 135\u0026nbsp;million in 1995, with projections estimating nearly 300\u0026nbsp;million cases by 2025 \u003csup\u003e3\u003c/sup\u003e. This rising prevalence places a significant financial strain on healthcare systems, particularly in low-income regions \u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. Diabetes is classified into two main types: type I, which affects 5\u0026ndash;10% of individuals and results from impaired insulin production, and type II, which accounts for approximately 90% of cases and arises from insulin resistance \u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. Persistent hyperglycemia, a hallmark of diabetes, can lead to severe complications, including damage to the central nervous system and blood vessels. Currently, several synthetic antidiabetic drugs, including meglitinides, biguanides, incretin mimetics, thiazolidinediones, sodium-glucose co-transporter 2 (SGLT2) inhibitors, and dipeptidyl peptidase-IV inhibitors, are under investigation for diabetes management \u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e. While these medications have shown promising therapeutic effects, they are often associated with adverse side effects such as headaches, diarrhea, hypoglycemia, impotence, and vision impairment \u003csup\u003e\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. Given these challenges, there is a growing interest in developing safer, more effective antidiabetic treatments derived from natural sources. Traditional medicines, particularly herbal plants and their extracts have been recognized as viable alternatives to synthetic drugs due to their safety and efficacy. Notably, approximately 90% of modern medications are directly or indirectly derived from plant-based compounds, reinforcing their potential for new drug development \u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e. Continued research on plant-based antidiabetic compounds is essential to discovering novel therapeutic agents with minimal side effects and enhanced clinical benefits \u003csup\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eAlzheimer\u0026rsquo;s disease (AD) is a progressive neurodegenerative disorder that significantly impairs the central nervous system \u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e. Several factors contribute to AD pathogenesis, including oxidative stress, amyloid plaque accumulation, neuroinflammation, and cholinergic dysfunction \u003csup\u003e\u003cspan additionalcitationids=\"CR12 CR13\" citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e. While the exact cause of AD remains unclear, and no definitive cure has been identified, disease management focuses on addressing these underlying factors to slow its progression and alleviate symptoms \u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e. Current treatments for AD do not provide a permanent cure but aim to mitigate cognitive decline, regulate behavioral symptoms, and improve overall quality of life \u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e. Despite advancements in therapeutic strategies, further research is essential to develop more effective treatments. One key aspect of AD pathology is the disruption of cholinergic neurotransmission, which is crucial for normal cognitive function. Acetylcholine (ACh), a vital neurotransmitter, is degraded by the enzyme acetylcholinesterase (AChE), leading to impaired synaptic transmission and exacerbating AD symptoms \u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. AChE inhibitors have emerged as a promising therapeutic approach, as they help maintain acetylcholine levels in the brain, thereby improving cognitive function \u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e. Numerous studies have highlighted the potential of plant-derived compounds in exhibiting anti-cholinesterase activity. Despite significant progress in pharmaceutical advancements, natural products continue to play a crucial role in drug discovery and healthcare. Increasing recognition of the therapeutic potential of medicinal plants has prompted pharmaceutical companies to explore plant-derived compounds as potential candidates for novel drug development \u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eInflammation is a complex biological response that plays a critical role in the development and progression of various pathological conditions, including arthritis and cardiovascular diseases \u003csup\u003e\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e. Current treatment strategies primarily rely on nonsteroidal anti-inflammatory drugs (NSAIDs), which inhibit cyclooxygenase (COX) enzymes. There are two COX isoforms: cyclooxygenase-1 (COX-1), which is constitutively expressed in most cells under normal physiological conditions, and cyclooxygenase-2 (COX-2), which is induced by pro-inflammatory mediators such as tumor necrosis factor-α (TNF-α), lipopolysaccharides (LPS), and tumor-promoting factors \u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e. Efforts to develop alternative anti-inflammatory agents have led to the identification of both natural and synthetic dual COX-2/5-lipoxygenase (5-LOX) inhibitors, which exhibit promising anti-inflammatory activity. Research has focused on understanding the structural and pharmacological properties of natural compounds and heterocyclic systems that function as dual COX-2/5-LOX inhibitors \u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e. However, a significant limitation of COX inhibition is its potential to stimulate 5-LOX, leading to increased leukotriene (LT) production. Although 5-LOX inhibitors have demonstrated protective effects, their widespread use is restricted due to various drawbacks \u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e. The adverse effects associated with COX and 5-LOX inhibitors have thus limited their application in treating inflammatory disorders \u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e. Given these challenges, there is an urgent need for new therapeutic strategies that effectively modulate inflammation while minimizing side effects. Celtis caucasica, commonly known as the Caucasian hackberry or Caucasian nettle tree, belongs to the Cannabaceae family. This medium-to-large deciduous tree is native to regions extending from Turkey to Central Asia and Assam, thriving in temperate climates \u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e. Notable for its longevity and adaptability to rocky ecosystems and various environmental stressors, \u003cem\u003eC. caucasica\u003c/em\u003e has been traditionally used in indigenous medicine. Its seeds and fruits have been employed to treat digestive ailments, including diarrhea and stomach disorders. Given its medicinal potential, further research into \u003cem\u003eC. caucasica\u003c/em\u003e may uncover novel bioactive compounds with anti-inflammatory properties, contributing to the development of safer and more effective therapeutic alternatives.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eChemicals Instruments used\u003c/h2\u003e \u003cp\u003eThe solvents employed in this study included ethanol, n-hexane, ethyl acetate, dichloromethane, chloroform (Master Chemical Supplier, Karachi, Pakistan), and distilled water (Islamia College Peshawar). Laboratory equipment utilized comprised glass funnels, filter papers, a thermostatically controlled water bath (STD/GMP), a digital magnetic stirrer (Model H3760-S), a vacuum rotary evaporator (Model RE-111, Labstac LLC, Pittsfield, MA, USA), and an analytical balance (Shimadzu, Karachi, Pakistan). GC-MS analysis was conducted using a Thermo GC Ultra (Version 5.0) system, coupled with a Thermo MS DSQ II and a GC 7890B-MS 5977B system.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003ePlant Collection and Identification\u003c/h3\u003e\n\u003cp\u003eThe fresh leaves of \u003cem\u003eC. caucasica\u003c/em\u003e were collected in November from Islamia College Peshawar. The specimen was identified taxonomically by Dr. Naveed Akhtar, Associate Professor at the Department of Botany, Islamia College Peshawar. A voucher specimen (ICP/Bot-102) was deposited in the Department of Botany for reference. The leaves were then cut into small pieces and dried in the shade in a well-ventilated area. After drying, the leaves were ground into a coarse powder using a pestle and mortar to facilitate the maceration process. Approximately 1 kg of the powdered material was used for subsequent laboratory procedures \u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003ch3\u003ePreparation of the crude extract\u003c/h3\u003e\n\u003cp\u003eOne kilogram of powdered \u003cem\u003eC. caucasica\u003c/em\u003e leaves was macerated in an appropriate volume of ethanol in a stainless-steel container, which was sealed and kept at room temperature for twelve days. To prevent solvent evaporation, the vessel was covered with a lid. The mixture was stirred three to four times a day using glass rods to ensure maximum extraction efficiency. After twelve days, the extract was first filtered through muslin cloth and then through Whatman No. 1 filter paper to remove plant residues. The filtrate was transferred to a clean container and concentrated under rotary vacuum at 50\u0026deg;C. The resulting extract was further dried in a water bath at 50\u0026deg;C, yielding 10 grams of dried crude ETCC.\u003c/p\u003e\n\u003ch3\u003eFractionation\u003c/h3\u003e\n\u003cp\u003eTen grams of crude ETCC was divided into two equal portions. One portion (5 g) was used as the crude methanolic extract, while the other portion underwent fractionation. The fractionation process began by dissolving the ETCC in distilled water. n-Hexane was then added to the mixture in a separating funnel, gently shaken, and left to settle for 10 minutes to separate the two phases: the n-hexane fraction and the aqueous layer. The n-hexane fraction was collected into a separate flask, and the remaining aqueous portion was further fractionated using a series of solvents, including dichloromethane, chloroform, n-hexane, and ethyl acetate, following the solvent-solvent fractionation method \u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e. Each solvent-extracted fraction was concentrated using a rotary evaporator at appropriate temperatures. This process resulted in the isolation of the crude ETCC and its different solvent fractions: NHFCC, dichloromethane fraction (DCMFCC), ethyl acetate fraction (EAFCC), chloroform fraction (CHCC), and n-butanol fraction. In this study, we conducted phytochemical analysis of the ETCC, NHFCC, and CHCC fractions.\u003c/p\u003e\n\u003ch3\u003eGC-MS analysis\u003c/h3\u003e\n\u003cp\u003eThe phytochemical composition of \u003cem\u003eC. caucasica\u003c/em\u003e extracts, including the ETCC and CHFCC, was determined using GC-MS. The analysis was performed on a Thermo GC Ultra (Version 5.0) coupled with a Thermo MS DSQ II and a GC 7890B-MS 5977B system. A TR-5MS capillary column (30 m length, 0.25 \u0026micro;m film thickness, 0.25 mm internal diameter) was used for compound separation. High-purity helium gas (99.99%) served as the carrier at a flow rate of 1 mL/min. The injector was set to split mode at 250\u0026deg;C, and 1 \u0026micro;L of the sample was injected for analysis. The oven temperature program started at 50\u0026deg;C, held for 2 minutes, and then increased to 150\u0026deg;C at a rate of 8\u0026deg;C/min. The temperature was further raised to 300\u0026deg;C at 15\u0026deg;C/min and held for 5 minutes to ensure optimal separation of the compounds (Cupido et al., 2022).\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eFTIR analysis\u003c/h2\u003e \u003cp\u003eFTIR analysis was conducted to identify the functional groups and structural changes in the molecules. Two grams of both the ETCC and CHFCC of \u003cem\u003eC. caucasica\u003c/em\u003e were used for the analysis. To prepare the FTIR-grade KBr, 50 mg was mixed with 1 mg of the ETCC and CHFCC samples. The dried samples of each extract were then placed on the FTIR spectrometer, with a resolution of 4 cm\u0026thinsp;\u0026minus;\u0026thinsp;1 and a scanning range of 4000\u0026ndash;400 cm\u0026thinsp;\u0026minus;\u0026thinsp;1 \u003csup\u003e1\u003c/sup\u003e.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eα-amylase Inhibitory Activity Assay\u003c/h3\u003e\n\u003cp\u003eTo assess the effect of \u003cem\u003eC. caucasica\u003c/em\u003e extracts and column semi-purified fractions on α-amylase activity in an enzyme-starch system \u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e, the following procedure was followed. A 1% concentration of each extract and semi-purified fraction of \u003cem\u003eC. caucasica\u003c/em\u003e was mixed with 25 mL of a 4% potato starch solution in a beaker. Then, 100 mg of α-amylase was added to the starch solution and thoroughly mixed. The reaction mixture was incubated at 37\u0026deg;C for 60 minutes to allow enzymatic activity. After the incubation period, 0.1 M NaOH was added to stop the reaction. The mixture was then centrifuged at 3000 \u0026times;g for 15 minutes, and the supernatant was carefully decanted. Glucose concentration was subsequently measured to assess enzyme activity.\u003c/p\u003e\n\u003ch3\u003eα-glucosidase Inhibitory Activity Assay\u003c/h3\u003e\n\u003cp\u003eThe α-glucosidase inhibitory activity of \u003cem\u003eC. caucasica\u003c/em\u003e extracts and column semi-purified fractions was evaluated using the method described by Pistia-Brueggeman and Hollingsworth (2001), as reported by \u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e. Inhibitory activity was assessed by incubating the extracts and fractions (62.5\u0026ndash;1000 \u0026micro;g/mL) with a mixture containing 10 \u0026micro;L of α-glucosidase (1 U/mL) and 125 \u0026micro;L of phosphate buffer (0.1 M, pH 6.8) at 37\u0026deg;C for 20 minutes. The enzyme reaction was initiated by adding 20 \u0026micro;L of a 1 M p-nitrophenyl-β-D-glucopyranoside (pNPG) substrate solution, and the mixture was further incubated for 30 minutes under the same conditions. The reaction was halted by the addition of 50 \u0026micro;L of 0.1 N sodium carbonate (Na₂CO₃). Absorbance was measured at 405 nm using a UV-Vis spectrophotometer, both for the sample and control. The percentage of α-glucosidase inhibition was calculated using the formula:\u003c/p\u003e \u003cp\u003eInhibitory Activity (%) \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{\\text{O}\\text{D}\\left(\\text{c}\\text{o}\\text{n}\\text{t}\\text{r}\\text{o}\\text{l}\\right)-\\text{O}\\text{D}\\left(\\text{s}\\text{a}\\text{m}\\text{p}\\text{l}\\text{e}\\right)}{\\text{O}\\text{D}}\\left(\\text{c}\\text{o}\\text{n}\\text{t}\\text{r}\\text{o}\\text{l}\\right))\\times\\:100\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003cp\u003eInhibitory Activity (%) \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{\\text{A}\\text{b}\\text{s}\\text{C}-\\text{A}\\text{b}\\text{s}\\text{C}}{\\text{A}\\text{b}\\text{s}\\text{C}}\\times\\:100\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003eIn-Vitro\u003c/b\u003e \u003cb\u003eAnti-Cholinesterase Activity\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe inhibitory activities of AChE and BChE were assessed using a spectrophotometric method based on Ellman's assay. ETCC and its EAFCC and CHCC fractions were tested at concentrations of 62.5, 125, 250, 500, and 1000 \u0026micro;g/mL to evaluate their ability to inhibit AChE and BChE. The assay involves the enzymatic hydrolysis of acetylthiocholine iodide by the respective cholinesterase enzyme, resulting in the production of thiocholine. This liberated thiocholine reacts with Ellman's reagent, 5,5\u0026prime;-dithiobis-2-nitrobenzoic acid (DTNB), to form a chromogenic product that can be quantitatively measured spectrophotometrically. The end products of the reaction are 5-thio-2-nitrobenzoate and 2-nitrobenzoate-5-mercaptothiocholine. The absorbance of 5-thio-2-nitrobenzoate was measured at 412 nm using a spectrophotometer. Galantamine was used as a positive control, and plant extracts were assessed at the same concentrations. All samples were incubated at 37\u0026deg;C for 20 minutes before absorbance readings were taken. The change in absorbance over time was used to calculate enzyme inhibition \u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e,\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe percentage of inhibition was calculated using the following formula:\u003c/p\u003e \u003cp\u003eEnzyme inhibition (%)\u0026thinsp;=\u0026thinsp;100\u0026thinsp;\u0026minus;\u0026thinsp;percent enzyme activity\u003c/p\u003e \u003cp\u003ePercent enzyme activity (%)\u0026thinsp;=\u0026thinsp;100 \u0026times; V/V\u003csub\u003emax\u003c/sub\u003e\u003c/p\u003e \u003cp\u003ewhere (V\u003csub\u003emax\u003c/sub\u003e) is an enzyme activity in the absence of an inhibitor.\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eCOX-2 Inhibitory Assay\u003c/h2\u003e \u003cp\u003eAn in vitro assay was conducted to evaluate the anti-inflammatory activity of \u003cem\u003eC. caucasica\u003c/em\u003e leaf extracts and column semi-purified fractions by assessing their inhibitory effect on cyclooxygenase-2 (COX-2) activity. The procedure outlined by \u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e, was followed. A COX-2 enzyme solution was prepared at a concentration of 300 U/mL and chilled on ice for 5 to 10 minutes to activate the enzyme. Co-factors, including hematin (1 mM), N,N,N\u0026prime;,N\u0026prime;-tetramethyl-p-phenylenediamine (TMPD, 0.24 mM), and glutathione (0.9 mM), were dissolved in a Tris-HCl buffer (0.1 M, pH 8) and mixed with the activated enzyme solution. The plant extracts and fractions were added at concentrations ranging from 62.5 to 1000 \u0026micro;g/mL and incubated at 25\u0026deg;C for 5 minutes. To initiate the enzymatic reaction, 20 \u0026micro;L of a 30 mM arachidonic acid solution was introduced, and the mixture was incubated for an additional 4 to 5 minutes. Celecoxib was used as a positive control. The absorbance of the reaction mixture was measured at 540 nm using a UV-Vis spectrophotometer.\u003c/p\u003e \u003cp\u003eInhibitory Activity (%) \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{\\text{A}\\text{b}\\text{s}\\text{C}-\\text{A}\\text{b}\\text{s}\\text{C}}{\\text{A}\\text{b}\\text{s}\\text{C}}\\times\\:100\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003cp\u003eHere: AbsC\u0026thinsp;=\u0026thinsp;absorbance of control, while AbsS\u0026thinsp;=\u0026thinsp;absorbance of sample [extract/fractions].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eLOX Inhibitory Assay\u003c/h2\u003e \u003cp\u003eIn addition to the COX-2 assay, a 5-lipoxygenase (5-LOX) inhibitory assay was used to evaluate the in-vitro anti-inflammatory activity of \u003cem\u003eC. caucasica\u003c/em\u003e extracts and semi-purified fractions, following the procedure described by \u003csup\u003e\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e\u003c/sup\u003e. Extracts and fractions were prepared at concentrations ranging from 62.5 to 1000 \u0026micro;g/mL. A 5-LOX enzyme solution was prepared at 10,000 U/mL, and the reaction was initiated by adding 80 mM linoleic acid as the substrate. The reaction mixture consisted of 250 \u0026micro;L of crude extract and 50 mM phosphate buffer (pH 6.3), to which 250 \u0026micro;L of enzyme solution was added. After 5 minutes of incubation, the substrate solution was introduced, and the mixture was well-mixed. Absorbance of both test and control samples was measured at 234 nm using a UV-Vis spectrophotometer. A concentration-response curve was constructed to determine the correlation between extract concentration and enzyme inhibition. The IC₅₀ value, representing the concentration inhibiting 50% of the enzyme activity, was calculated, and the percentage inhibition of 5-LOX activity was determined.\u003c/p\u003e \u003cp\u003eInhibitory Activity (%) \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{\\text{A}\\text{b}\\text{s}\\text{C}-\\text{A}\\text{b}\\text{s}\\text{C}}{\\text{A}\\text{b}\\text{s}\\text{C}}\\times\\:100\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003cp\u003eHere: AbsC\u0026thinsp;=\u0026thinsp;absorbance of control, while AbsS\u0026thinsp;=\u0026thinsp;absorbance of sample (extract/fractions).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eDDPH Free Radical Scavenging Assay\u003c/h2\u003e \u003cp\u003eThe antioxidant activity of ETCC, NHFCC, and CHCC, and the standard compound was evaluated based on free radical scavenging activity against the stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, following the method described by \u003csup\u003e\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e,\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e. The plant extracts and fractions were dissolved in ethanol, and ascorbic acid was used as a standard at concentrations ranging from 1 to 100 \u0026micro;g/mL. A 0.002% DPPH solution was prepared in methanol, and 1 mL of this solution was added to 1 mL of each plant extract/fraction and standard solution in separate test tubes. A blank control consisted of 1 mL of methanol and 1 mL of the 0.002% DPPH solution. The reaction mixtures were incubated at room temperature in the dark for 30 minutes, after which absorbance was measured at 517 nm using a UV-Vis spectrophotometer.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eABTS Free Radical Scavenging Assay\u003c/h2\u003e \u003cp\u003eThe antioxidant capacity of the compound was evaluated using the ABTS (2,2'-azinobis [3-ethylbenzthiazoline]-6-sulphonic acid) free radical assay, as described by \u003csup\u003e\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u003c/sup\u003e. In this method, antioxidants scavenge radical cations of ABTS, reducing absorbance at 734 nm. To prepare the ABTS solution, potassium persulfate (2.45 mM) and ABTS (7 mM) were mixed and incubated in the dark at room temperature for 12\u0026ndash;16 hours to generate the radical cation solution. For the assay, the ABTS radical solution was diluted with phosphate buffer (0.01 M, pH 7.4) to achieve an absorbance of 0.70 at 734 nm. A mixture of 300 \u0026micro;L of the compound and 3.0 mL of the ABTS solution was added to a cuvette to measure the scavenging activity. The absorbance was recorded at 734 nm after 1 minute and again after 6 minutes of mixing. Ascorbic acid was used as a positive control. The assay was performed in triplicate, and the percentage of inhibition was calculated using the formula provided by \u003csup\u003e\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis:\u003c/h2\u003e \u003cp\u003eData are presented as the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM, with significance determined using one-way ANOVA followed by Bartlett's test. Asterisks (*) denote statistically significant differences compared to the control group. A p-value of \u0026le;\u0026thinsp;0.05 was considered statistically significant. All samples were evaluated in triplicate.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eGC-MS analysis\u003c/h2\u003e \u003cp\u003eGC-MS analysis was conducted to characterize the phytochemical composition of the crude ETCC and its CHFCC. The mass spectra were compared with reference patterns from the National Institute of Standards and Technology (NIST) database, which includes over 62,000 compounds. Unidentified constituents (Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) were matched with known spectra from the NIST library to determine their molecular weight, structure, and chemical identity.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003ePhytochemical composition of ETCC\u003c/h2\u003e \u003cp\u003ePhytochemical analysis of the ethanol extract of \u003cem\u003eC. caucasica\u003c/em\u003e using GC-MS identified sixteen compounds belonging to various chemical classes. Many of these compounds are known for their significant bioactivities, including antioxidant, anti-inflammatory, anti-Alzheimer\u0026rsquo;s, and antimicrobial properties. The identified compounds include 4-O-Methylmannose (C₇H₁₄O₆) at RT 13.571, Thiophene, tetrahydro-2-methyl- (C₅H₁₀S) at RT 14.023, Benzoic acid, 2-ethylhexyl ester (C₁₅H₂₂O₂) at RT 14.602, Guaifenesin (C₁₀H₁₄O₄) at RT 15.243, Hexadecanoic acid, methyl ester (C₁₇H₃₄O₂) at RT 16.911, n-Hexadecanoic acid (C₁₆H₃₂O₂) at RT 17.264, Hexadecanoic acid, ethyl ester (C₁₈H₃₆O₂) at RT 17.577, 9,12-Octadecadienoic acid (Z,Z)-, methyl ester (C₁₉H₃₄O₂) at RT 18.458, Phytol (C₂₀H₄₀O) at RT 18.710, 12,15-Octadecatrienoic acid (Z,Z,Z)- (C₁₈H₃₀O₂) at RT 18.847, Oleic acid (C₁₈H₃₄O₂) at RT 18.881, Octadecanoic acid (C₁₈H₃₆O₂) at RT 19.115, Eicosanoic acid (C₂₀H₄₀O₂) at RT 20.825, and 1,9-Tetradecadiene (C₁₄H₂₆) at RT 22.148 (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePhytochemical composition of ETCC\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"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=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e%Area\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eName of Compound\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMF\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMW\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003ePeak Area\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eLibrary\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13.571\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4-O-Methylmannose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e7\u003c/sub\u003eH\u003csub\u003e14\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e194.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e13627571\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14.023\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eThiophene, tetrahydro-2-methyl-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e5\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e102.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e70158237\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14.602\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBenzoic acid, 2-ethylhexyl ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eG\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e234.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e10061527\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15.243\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGuaifenesin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e10\u003c/sub\u003eH\u003csub\u003e14\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e198.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e6218972\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16.911\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHexadecanoic acid, methyl ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e270.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e7878301\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e17.264\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003en-Hexadecanoic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e32\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e256.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e31846853\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e18710\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePhytol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e20\u003c/sub\u003eH\u003csub\u003e40\u003c/sub\u003eO\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e296.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e18433814\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e18.847\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12,15-Octadecatrienoic acid, (Z,Z,Z)-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e278.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e25176956\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e18.881\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOleic Acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e82.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e39728445\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e19.115\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOctadecanoic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e36\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e284.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e28237421\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e22.148\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1,9-Tetradecadiene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e14\u003c/sub\u003eH\u003csub\u003e26\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e194.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5674829\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e24.558\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSqualene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e30\u003c/sub\u003eH\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e410.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e13437612\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e26.868\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ebeta.-Tocopherol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e48\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e416.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e7857583\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e28.338\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eVitamin E\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e8\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e176.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e67832529\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e31.912\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003egamma.-Sitosterol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e29\u003c/sub\u003eH\u003csub\u003e50\u003c/sub\u003eO\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e414.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e41326518\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e35.691\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBenzoic acid, 2-(2,4-dihydroxybenzylidenamino)-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e14\u003c/sub\u003eH\u003csub\u003e11\u003c/sub\u003eNO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e257.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e43807647\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003ePhytochemical composition of CHFCC\u003c/h2\u003e \u003cp\u003eSimilarly, GC-MS analysis of the CHFCC confirmed the presence of approximately twenty compounds, as depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. These compounds include Phthalic acid, di(2-propylpentyl) ester (11.94%), Octadec-9-enoic acid (11.76%), n-Hexadecanoic acid (6.38%), p-Xylene (5.29%), Ethylbenzene (4.90%), Benzene, nitro (2.81%), Nonacosane (6.20%), 9-Octadecenoic acid (Z)-, methyl ester (1.56%), and Benzoic acid, 2-ethylhexyl ester (1.07%), among others.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePhytochemical composition of CHFCC\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"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=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e%Area\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eName of Compound\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMF\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMW\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003ePeak Area\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eLibrary\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2.356\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTrichloromethane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCHCl₃\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e119.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3096842\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2.551\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eEthylbenzene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₈H₁₀\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e106.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e9382483\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2.658\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ep-Xylene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e8\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e106.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e10128672\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2.725\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTrichloromethane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCHCl₃\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e119.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2485285\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2.959\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ep-Xylene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e8\u003c/sub\u003eH10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e106.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3781301\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5.837\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBenzene, nitro\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e5\u003c/sub\u003e NO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e123.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5372028\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12.356\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePhenol,2,5-bis(1,1-dimethylethyl)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC14H220\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e206.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1526303\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12.525\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eDodecanoic acid, methyl ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₁₃H₂₆O₂\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e214.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1618672\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14.606\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBenzoic acid, 2-ethylhexyl ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₁₅H₂₂O₂\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e234.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2048960\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e17.277\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003en-Hexadecanoic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₁₆H₃₂O₂\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e256.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e12213770\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e18.465\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMethyl 10-trans,12-cis-octadecadienoate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₁₉H₃₆O₂\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e296.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2057046\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e18.547\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e9-Octadecenoic acid (Z)-, methyl ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₁₉H₃₆O₂\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e296.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2991241\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e18.885\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e11.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOctadec-9-enoic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₁₈H₃₄O₂\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e282.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e22501628\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e19.124\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOctadecanoic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₁₈H₃₆O₂\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e284.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e10152548\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e22.105\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e11.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePhthalic acid, di(2-propylpentyl) ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₂₆H₄₂O₄\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e410.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e22847404\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e24.559\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSqualene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₆₀H₁₀\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e410.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5412358\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e25.503\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNonacosane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₂₉H₆₀\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e404.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e11859419\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e26.878\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003edelta. -Tocopherol, O-methyl-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₂₉H₄₈O₃\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e440.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e28.349\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eVitamin E\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₂₉H₄₈O₂\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e430.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e35257222\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e31.909\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ebeta. -Sitosterol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC₂₈H₄₆O\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e414.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e13050602\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNIST20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eFT-IR analysis of ETCC\u003c/h2\u003e \u003cp\u003eThe FTIR analysis of the \u003cem\u003eC. caucasica\u003c/em\u003e ethanol extract identified several functional groups, including aromatic compounds, alcohols, amides, methylene groups, alkanes, ketones, and aldehydes. Notably, the majority of the peaks corresponded to alkane groups, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e and Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \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\u003eFTIR analysis of \u003cem\u003eC. caucasica\u003c/em\u003e leaves ETCC.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePeak Number\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eWavenumber (cm-1)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIntensity\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eFunctional group\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eCompound identifier\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e611.28341\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e62.91267\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;H bending\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003earomatic compounds\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e879.65173\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e65.15021\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;H bending\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003earomatic compounds\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1028.74524\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e37.02025\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;O stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ealcohols, ethers, or esters\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1088.38265\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e66.97643\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;O stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ealcohols, ethers, or esters.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1326.93227\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e87.81577\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;N stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eamide groups\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1379.115\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e81.8381\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC-H bending vibration\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003emethyl group\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1416.38838\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e82.50645\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC-H bending\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003emethyl or methylene group\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1449.93442\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e82.78508\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;H Bending\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAlkanes\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1654.938\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e92.89867\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026thinsp;=\u0026thinsp;O stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eketones, aldehydes, amides, or carboxylic acids\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2832.77676\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e86.17968\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eH\u0026ndash;C\u0026thinsp;=\u0026thinsp;O stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ealdehydes\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2937.14222\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e82.16133\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;H Stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003emethylene\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2974.41559\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e75.88517\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;H stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ealkanes\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3313.60334\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e68.73231\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eO\u0026ndash;H stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAlkanes\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003eFT-IR analysis of CHFCC\u003c/h2\u003e \u003cp\u003eThe FTIR analysis of the CHFCC revealed characteristic absorption peaks, as detailed in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e of the supplementary material. The wavelength values in the FTIR spectra correspond to specific chemical bonds within the molecule, which are identified through IR absorption. The chloroform extract of \u003cem\u003eC. caucasica\u003c/em\u003e contains functional groups such as metal-organic complexes, alkanes, methylene groups, alcohol esters, and aromatic compounds. Notably, the most prominent peak was associated with aromatic compounds in the FTIR analysis.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eFTIR analysis of \u003cem\u003eC. caucasica\u003c/em\u003e leaves CHFCC.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePeak Number\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eWavenumber (cm-1)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIntensity\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eFunctional group\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eCompound Identifier\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e469.64457\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e91.96578\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003emetal-ligand stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eNot identified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e667.19347\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e42.67843\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC-H bending\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003earomatic compounds\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e745.46757\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.47095\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;H bending\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003earomatic compounds\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e928.10712\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e96.53538\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;H bending\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003earomatic compounds\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1028.74524\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e95.82001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;O stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ealcohols, ethers, or esters\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1215.11213\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e48.30565\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;O stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eesters, ethers, or alcohols\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1423.84305\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e97.83264\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC-H bending\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003emethyl group\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1520.75384\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e98.59872\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;C stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003earomatic compounds\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1602.75527\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e99.25807\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;C stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003earomatic compounds\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2400.40557\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e99.83959\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u0026ndash;H stretching\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ealkynes\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3019.14365\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e95.66922\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMetal-ligand vibrations\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003emetal-organic complexes or coordination compounds\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 \u003cb\u003eIn-Vitro\u003c/b\u003e \u003cb\u003eAnti-Cholinesterase Potential\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe ETCC, the n-hexane fraction (NHFCC), and the CHFCC demonstrated significant inhibitory effects against AChE and BChE at varying concentrations, as shown in Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e. The highest inhibition of AChE was observed at 500 \u0026micro;g/mL for CHFCC, with an inhibition of 86.44% and an IC50 value of 13.2. For BChE at the same concentration, CHFCC exhibited a 92.67% inhibition with an IC50 of 9.66. In comparison, ETCC showed 79.49% inhibition (IC50 16.5) against AChE and 82.61% inhibition (IC50 21.9) against BChE. NHFCC displayed 79.48% (IC50 27.4) inhibition of AChE and 79.37% (IC50 21.2) inhibition of BChE, respectively.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePercent inhibition of AChE and BChE by ETCC, NHFCC and CHFCC\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSolvents\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eConcentration (\u0026micro;g/ml)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAChE percent inhibition\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e (\u0026micro;g/ml)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e% BChE inhibition\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e (\u0026micro;g/ml)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eETCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e79.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e16.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e82.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e21.9\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e75.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e77.60\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e72.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72.83\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e65.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e57.55\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e54.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e54.58\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eNHFCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e79.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e27.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e79.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e21.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e75.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e74.72\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e70.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e70.47\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e57.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e65.50\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e51.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e48.46\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eCHFCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e86.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e13.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e92.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e9.66\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e81.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e88.58\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e79.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e82.54\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e74.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e78.20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e52.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e73.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eGalantamine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e94.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e6.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e93.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e3.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e85.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e86.45\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e80.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e80.58\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e76.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e75.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e71.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e70.80\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\u003eThe data are presented as the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of the mean (SEM). One-way ANOVA followed by Bartlett's test was used to assess significance. Asterisks denote the level of significance: \"\" (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) indicates highly significant inhibition compared to the control, \"\" (p\u0026thinsp;\u0026lt;\u0026thinsp;0.01) represents moderate significance, \"\" (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) suggests mild significance, and \"ns\" (not significant) denotes no significant inhibition at certain concentrations.\u003c/p\u003e \u003cp\u003e \u003cb\u003eIn-Vitro\u003c/b\u003e \u003cb\u003eAnti-Diabetic Potential\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe anti-diabetic inhibitory activity of \u003cem\u003eC. caucasica\u003c/em\u003e extracts against α-glucosidase and α-amylase was evaluated to assess their potential antidiabetic effects (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). Among the extracts, CHFCC exhibited the highest inhibition of α-glucosidase (88.61% at 500 \u0026micro;g/mL) with an IC₅₀ of 11.99 \u0026micro;g/mL, followed by NHFCC (83.51%, IC₅₀ = 19.48 \u0026micro;g/mL) and ETCC (79.37%, IC₅₀ = 18.98 \u0026micro;g/mL). The standard drug acarbose showed the highest inhibition of α-glucosidase (94.08%) with an IC₅₀ of 11.98 \u0026micro;g/mL. In terms of α-amylase inhibition, CHFCC demonstrated the strongest activity (91.36% at 500 \u0026micro;g/mL) with an IC₅₀ of 4.22 \u0026micro;g/mL, followed by NHFCC (88.52%, IC₅₀ = 14.67 \u0026micro;g/mL) and ETCC (77.40%, IC₅₀ = 12.92 \u0026micro;g/mL). Acarbose, the control, showed the most potent inhibitory effect (97.23%) with an IC₅₀ of 9.72 \u0026micro;g/mL.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePercent inhibition of alpha glucosidase and alpha amylase by ETCC, NHFCC and CHFCC.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSolvents\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eConcentration (\u0026micro;g/mL)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAlpha glucosidase percent inhibition\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e (\u0026micro;g/mL)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAlpha amylase inhibition\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e (\u0026micro;g/mL)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eCHFCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e88.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e11.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e91.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e14.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e82.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e85.34\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e75.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e81.39\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e69.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e76.47\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e62.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e71.44\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eETCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e79.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e18.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e77.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e12.92\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e73.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72.57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e69.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e67.36\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e63.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e62.56\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e53.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e57.37\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eNHFCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e83.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e19.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e88.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e14.67\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e75.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e82.4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e67.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e74.54\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e63.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e67.34\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e56.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e61.30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eAcarbose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e94.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e11.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e97.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e9.72\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e87.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e92.45\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e81.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e85.90\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e76.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e81.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e71.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e76.90\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 \u003cb\u003eIn-Vitro\u003c/b\u003e \u003cb\u003eAnti-Inflammatory Potential\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe anti-inflammatory activity of various \u003cem\u003eC. caucasica\u003c/em\u003e extracts was assessed by evaluating their inhibitory effects on cyclooxygenase-2 (COX-2), cyclooxygenase-1 (COX-1), and 5-lipoxygenase (5-LOX) (Table\u0026nbsp;\u003cspan refid=\"Tab7\" class=\"InternalRef\"\u003e7\u003c/span\u003e). At a concentration of 500 \u0026micro;g/mL, CHFCC exhibited the highest inhibition of COX-2 (88.61%), followed by ETCC (79.08%) and NHFCC (75.32%). The standard drug, Celecoxib, demonstrated the highest inhibition of COX-2 (94.08%). Regarding COX-1 inhibition, CHFCC showed 69.58% inhibition, while NHFCC exhibited 61.30% inhibition. Celecoxib was found to be the most potent inhibitor of COX-1 among the samples. For 5-LOX inhibition, NHFCC demonstrated the highest activity (88.88%), followed by ETCC (79.37%) and CHFCC (73.80%). Montelukast, the positive control, showed the greatest 5-LOX inhibition (97.23%).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab7\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePercent inhibition of COX-2, COX-1 and 5-LOX by ETCC, NHFCC and CHFCC\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSolvents\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eConcentration (\u0026micro;g/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eCOX-2 percent inhibition\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eCOX-1 percent inhibition\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e5-LOX percent inhibition\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(\u0026micro;g/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e(\u0026micro;g/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e(\u0026micro;g/mL)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eCHFCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e88.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e23.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e69.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e139.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e73.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e106.97\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e82.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e62.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e66.94\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e75.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e55.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e59.72\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e69.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e49.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e51.84\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e62.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e41.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e43.80\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eETCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e79.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e43.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e54.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e673.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e79.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e60.68\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e72.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e45.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e72.37\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e65.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e37.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e65.30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e61.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e31.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e58.42\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e53.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e22.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50.52\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eNHFCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e75.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e77.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e61.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e286.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e88.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e32.84\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e67.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e55.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e83.54\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e62.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e49.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e75.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e55.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e42.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e67.68\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e34.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e59.82\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eCelecoxib\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"4\" rowspan=\"5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e94.08\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\" morerows=\"4\" rowspan=\"5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"4\" rowspan=\"5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e87.45\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e81.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11.98\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e76.40\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e71.80\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eMontelukast\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"4\" rowspan=\"5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e97.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"4\" rowspan=\"5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"4\" rowspan=\"5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e92.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e85.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.72\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e81.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e76.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eIndomethacin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"4\" rowspan=\"5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\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\u003e91.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e20.91\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\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\u003e85.34\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\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\u003e78.39\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\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\u003e72.47\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\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\u003e65.44\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003eDPPH and ABTS anti-radicals Assay\u003c/h2\u003e \u003cp\u003eThe antioxidant capacity of various extracts was evaluated using the DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2\u0026prime;-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) assays to assess their radical scavenging activities (Table\u0026nbsp;\u003cspan refid=\"Tab8\" class=\"InternalRef\"\u003e8\u003c/span\u003e). Among all the samples tested, CHFCC exhibited the highest antioxidant activity, with IC₅₀ values of 77.24 \u0026micro;g/mL (DPPH) and 60.67 \u0026micro;g/mL (ABTS). ETCC and NHFCC showed moderate and lower antioxidant activities, respectively. Ascorbic acid, the positive control, demonstrated the highest antioxidant activity, with the lowest IC₅₀ values of 2.81 \u0026micro;g/mL for DPPH and 4.6 \u0026micro;g/mL for ABTS.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab8\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 8\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAntioxidant activity of ETCC, CHFCC and NHCS.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCompound\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eConcentration (\u0026micro;g/mL)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eDPPH percent inhibition\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e (\u0026micro;g/mL)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eABTS percent inhibition\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e (\u0026micro;g/mL)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eCHFCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e75.32\u0026thinsp;\u0026plusmn;\u0026thinsp;2.87\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e77.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e79.36\u0026thinsp;\u0026plusmn;\u0026thinsp;1.03\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\" morerows=\"5\" rowspan=\"6\"\u003e \u003cp\u003e60.67\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e67.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.54\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e72.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.53\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e62.78\u0026thinsp;\u0026plusmn;\u0026thinsp;1.07\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e65.31\u0026thinsp;\u0026plusmn;\u0026thinsp;2.62\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e55.78\u0026thinsp;\u0026plusmn;\u0026thinsp;1.87\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e58.41\u0026thinsp;\u0026plusmn;\u0026thinsp;1.04\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e50.53\u0026thinsp;\u0026plusmn;\u0026thinsp;2.51\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eETCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e60.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e438.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e63.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 \u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e51.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e55.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.59 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e328.35\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e43.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.54 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e46.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e34.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.75 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e37.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e27.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.79 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e31.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eNHFCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e54.92\u0026thinsp;\u0026plusmn;\u0026thinsp;1.72 \u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e673.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e57.39\u0026thinsp;\u0026plusmn;\u0026thinsp;1.50 \u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e45.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.90 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e51.56\u0026thinsp;\u0026plusmn;\u0026thinsp;3.83 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e469.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e37.89\u0026thinsp;\u0026plusmn;\u0026thinsp;2.31 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e44.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.54 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e31.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e34.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.94 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e22.80\u0026thinsp;\u0026plusmn;\u0026thinsp;1.41 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e27.36\u0026thinsp;\u0026plusmn;\u0026thinsp;1.09 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eAscorbic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e95.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e2.81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e93.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.54\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e4.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e91.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e87.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e87.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.81\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e83.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e83.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e78.89\u0026thinsp;\u0026plusmn;\u0026thinsp;0.91\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e77.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e72.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19\u003csup\u003e*\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\u003eData are presented as the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM; one-way ANOVA followed by Bartlett's test was applied to determine significance; \"***\" indicates highly significant inhibition compared to control (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), observed in CHFCC. \"*\" denotes moderate significance (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05), seen in ETCC and NHFCC. \"ns\" (not significant) suggests no statistically relevant inhibition at certain concentrations.\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussions","content":"\u003cp\u003ePlant-based medicines have gained increasing popularity worldwide \u003csup\u003e\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e,\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e\u003c/sup\u003e, as researchers continually seek natural solutions that can be used to treat, diagnose, alleviate, or prevent various diseases \u003csup\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u003c/sup\u003e. Evaluating the therapeutic potential of these readily available natural compounds offers an opportunity to develop more affordable treatment options, especially in light of the current global economic challenges \u003csup\u003e\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u003c/sup\u003e. It is essential to recognize that enzyme inhibition in various biological compartments plays a crucial role in the pharmacodynamics of many drugs. In this study, fractions and extracts of \u003cem\u003eC. caucasica\u003c/em\u003e were investigated for their phytochemical composition, as well as their antioxidant, anti-cholinesterase, anti-inflammatory, and anti-diabetic activities. In the in vitro anti-diabetic assay, different extracts and fractions of \u003cem\u003eC. caucasica\u003c/em\u003e demonstrated significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) dose-dependent inhibition of α-amylase and α-glucosidase activity when compared to the standard (Acarbose, IC₅₀: 11.98 and 9.72 \u0026micro;g/mL). The IC₅₀ values for the extracts ranged from 88.61\u0026ndash;83.51 \u0026micro;g/mL for α-glucosidase and 91.36\u0026ndash;88.52 \u0026micro;g/mL for α-amylase inhibition, respectively. These findings are consistent with previous research by \u003csup\u003e\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/sup\u003e, which showed that \u003cem\u003eFicus benghalensis\u003c/em\u003e bark also had considerable inhibitory effects on α-glucosidase and α-amylase activity. Glucosidases are essential enzymes involved in various biological processes, such as the hydrolysis of dietary carbohydrates \u003csup\u003e\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u003c/sup\u003e. α-Glucosidase, found on the brush-border membrane of intestinal cells, catalyzes carbohydrate digestion \u003csup\u003e\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e\u003c/sup\u003e. The inhibitory effects of \u003cem\u003eF. benghalensis\u003c/em\u003e bark on α-glucosidase have been attributed to its phenolic compounds and flavonoid content \u003csup\u003e\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eIn this study, the fractions of \u003cem\u003eC. caucasica\u003c/em\u003e were evaluated for their anti-inflammatory properties through two enzymatic assays, namely COX-2 and 5-LOX inhibition. The research focused on assessing the anti-inflammatory activities of various \u003cem\u003eC. caucasica\u003c/em\u003e extracts based on their ability to inhibit COX-2, COX-1, and 5-LOX enzymes. The CHFCC extract exhibited the highest COX-2 inhibition at a concentration of 500 \u0026micro;g/mL, achieving 88.61% inhibition, which is comparable to the potency of the control drug, Celecoxib (94.08%). These results suggest that CHFCC contains potent COX-2 inhibitory compounds. Selective inhibition of COX-2 is crucial in anti-inflammatory therapy, as it helps avoid the adverse gastrointestinal effects often associated with COX-1 inhibition. Moderate COX-1 inhibition was observed with CHFCC (69.58%) and NHFCC (61.30%), indicating a selective inhibition pattern. These findings align with previous research, such as that of \u003csup\u003e\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e\u003c/sup\u003e, which reported a 25-fold selectivity for COX-2 over COX-1 in the ethanolic extract of Terminalia chebula fruit, with IC₅₀ values of 3.75 \u0026micro;g/mL for COX-2 and 90 \u0026micro;g/mL for COX-1. Regarding 5-LOX inhibition, NHFCC exhibited the highest activity (88.88%), suggesting the presence of bioactive compounds targeting the leukotriene pathway. This is consistent with studies on flavonoids, which are known to act as dual inhibitors of both COX-2 and 5-LOX, offering a comprehensive approach to inflammation treatment \u003csup\u003e\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e\u003c/sup\u003e. The dual inhibition of COX and 5-LOX pathways by \u003cem\u003eC. caucasica\u003c/em\u003e extracts is particularly significant, as such compounds are likely to exhibit enhanced anti-inflammatory effects while minimizing side effects associated with selective enzyme inhibition. For example, curcumin has been shown to have dual inhibitory activity against both 5-LOX and COX-1/2 enzymes \u003csup\u003e\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e\u003c/sup\u003e. The diverse inhibitory activities of \u003cem\u003eC. caucasica\u003c/em\u003e extracts against COX-2, COX-1, and 5-LOX highlight their potential as pharmaceutical anti-inflammatory agents. The strong inhibition of 5-LOX and excellent selectivity for COX-2 suggest that these extracts could be developed into safer and more effective anti-inflammatory therapies.\u003c/p\u003e \u003cp\u003eAChE is an enzyme that plays a critical role in terminating nerve impulses in cholinergic synapses by hydrolyzing the neurotransmitter ACh into choline and acetic acid, particularly within the blood and neurological systems \u003csup\u003e\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e. The findings from this study indicate that fractions of \u003cem\u003eC. caucasica\u003c/em\u003e exhibit strong inhibitory activity against AChE and BChE, both of which are implicated in neurodegenerative diseases like Alzheimer's disease. Among the fractions tested, CHFCC was the most effective natural inhibitor of cholinesterase. Other fractions, including ETCC and NHFCC, also showed significant inhibitory activity, further supporting the therapeutic potential of C. caucasica. These results suggest that further investigation into the bioactive compounds in these fractions could lead to the development of natural anti-Alzheimer's drugs. Previous studies support the notion that plant-derived compounds can effectively inhibit cholinesterase enzymes. For instance, isoquinoline alkaloids like sanguinarine and chelerythrine, found in Macleaya cordata extracts, have demonstrated effective BChE inhibition, with IC₅₀ values ranging from 25.74 to 28.04 \u0026micro;g/mL \u003csup\u003e\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e\u003c/sup\u003e. Additionally, methanolic extracts of several Ayurvedic medicinal herbs have been shown to exhibit AChE inhibitory activity, highlighting the potential of plant-based inhibitors in treating cognitive disorders \u003csup\u003e\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e\u003c/sup\u003e. The pronounced inhibitory action of CHFCC against both AChE and BChE suggests that \u003cem\u003eC. caucasica\u003c/em\u003e may contain bioactive compounds with dual inhibitory activity. Dual inhibition of these enzymes is especially beneficial in managing Alzheimer's disease, as both enzymes are involved in acetylcholine metabolism, and their concomitant inhibition could enhance cholinergic neurotransmission \u003csup\u003e\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e\u003c/sup\u003e. Moreover, in animal models, selective inhibition of BChE has been shown to elevate acetylcholine levels in the brain and improve cognitive function \u003csup\u003e\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe antioxidant system of an organism plays a critical role in scavenging free radicals produced during normal metabolic processes. However, an excess of free radicals can lead to oxidative stress \u003csup\u003e\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e\u003c/sup\u003e. Antioxidants, which can be either enzymatic or non-enzymatic, neutralize the toxic effects of these free radicals. Plants possess a variety of antioxidant compounds that make up their defense system. In this study, the antioxidant potential of \u003cem\u003eC. caucasica\u003c/em\u003e extracts (ETCC and CFCC) was assessed using the ABTS and DPPH scavenging assays. CHFCC demonstrated a high antioxidant potential, likely due to the presence of phenolic and flavonoid compounds, which are well known for their free radical scavenging abilities. Phenolic compounds exert their antioxidant activity by donating hydrogen atoms or electrons, stabilizing free radicals, and preventing damage caused by oxidative stress. This finding is consistent with studies on Cupressus species, which also exhibited substantial antioxidant capacity, particularly from ethyl acetate fractions, aligning with our observations for CHFCC \u003csup\u003e\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e\u003c/sup\u003e. ETCC exhibited moderate antioxidant activity, with IC₅₀ values of 438.38 \u0026micro;g/mL in the DPPH test and 328.35 \u0026micro;g/mL in the ABTS test. Literature suggests that the solvent used for extraction significantly impacts the yield and bioavailability of phenolic compounds \u003csup\u003e\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e\u003c/sup\u003e. While ETCC showed considerable antioxidant capacity, its lower efficiency may be attributed to differences in solvent-extractable polyphenolic content. NHFCC demonstrated the lowest antioxidant potential, with IC₅₀ values of 673.92 \u0026micro;g/mL (DPPH) and 469.02 \u0026micro;g/mL (ABTS), indicating a lower concentration of active antioxidant constituents. Similar trends have been observed in n-hexane extracts of medicinal plants, where non-polar fractions contain lower phenolic content and exhibit less radical scavenging activity \u003csup\u003e\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eTo further confirm the presence of these phytochemicals, GC-MS was employed. The GC-MS analysis revealed 16 major compounds in the ethanolic extract and 20 in the chloroform extract. Notable bioactive compounds identified in ETCC include 4-O-methylmannose, which exhibits antimicrobial activity \u003csup\u003e\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e\u003c/sup\u003e, and octadecadienoic acid (Z,Z)-methyl ester, which has antimicrobial, antioxidant, and anti-inflammatory properties. Other compounds such as hexadecanoic acid, methyl ester, and n-hexadecanoic acid exhibit a range of bioactive properties, including antioxidant, nematicidal, pesticidal, and anti-inflammatory activities \u003csup\u003e\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e,\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e\u003c/sup\u003e. Additionally, phytol, identified in the extract, has antimicrobial activity and is known for its stability and low toxicity \u003csup\u003e\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e\u003c/sup\u003e, while glycerin, found in CHFCC, has antibacterial properties \u003csup\u003e\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e\u003c/sup\u003e. Other compounds like thiophene, tetrahydro-2-methyl, and squalene have been reported for their antimalarial, cytotoxic, and antioxidant activities \u003csup\u003e\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e\u003c/sup\u003e. Oleic acid, another identified compound, has various bioactive properties, including anti-inflammatory, antiandrogenic, and cancer-preventive effects \u003csup\u003e\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eFourier-transform infrared (FTIR) spectroscopy is a valuable tool for identifying the types of chemical bonds and functional groups present in compounds. By analyzing the peak values in the infrared radiation region, the functional groups of active components can be identified, providing insights into the chemical composition of the sample. The FTIR spectrum of ETCC reveals a diverse array of phytochemicals, including flavonoids, terpenoids, and other polyphenolic compounds. These compounds are well-known for their antioxidant properties, which enable them to scavenge free radicals and alleviate oxidative stress \u003csup\u003e\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e\u003c/sup\u003e. Some flavonoids and terpenoids also exhibit antidiabetic properties, modulating carbohydrate metabolism and enhancing insulin sensitivity \u003csup\u003e\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e\u003c/sup\u003e. The presence of an amide group in the FTIR spectrum suggests the presence of alkaloids or peptides, which are associated with anticholinesterase activity and may be beneficial in treating neurodegenerative diseases \u003csup\u003e\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e\u003c/sup\u003e. Additionally, the prominence of aromatic compounds in CHFCC indicates a high concentration of phenolic constituents, known for their potent antioxidant activity. The antioxidant properties of phenolic compounds are primarily attributed to their hydroxyl groups, which donate hydrogen atoms to neutralize free radicals and mitigate oxidative stress \u003csup\u003e\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e\u003c/sup\u003e. Esters detected in the extract may contribute to its anti-inflammatory activity by modulating inflammatory processes. Some ester derivatives, such as those derived from ibuprofen, have been shown to possess enhanced anti-inflammatory properties compared to the parent compound, likely due to improved bioavailability and selective action \u003csup\u003e\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e\u003c/sup\u003e. Furthermore, the presence of metal-organic complexes suggests the potential existence of metalloproteins or coordination compounds. These complexes may play a role in various biochemical activities, such as enzymatic inhibition, which could contribute to anticholinesterase activity. Metalloproteins are involved in numerous physiological processes, and their interaction with metal ions is crucial for regulating enzymatic functions, including those involved in neurotransmission. Overall, FTIR analysis provides valuable insights into the chemical composition of ETCC, highlighting the presence of bioactive compounds that contribute to its antioxidant, anti-inflammatory, and potential anticholinesterase activities. These findings offer a deeper understanding of the therapeutic potential of \u003cem\u003eC. caucasica\u003c/em\u003e in treating oxidative stress, inflammation, and neurodegenerative diseases.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe current research presents a thorough phytochemical and pharmacological analysis of \u003cem\u003eC. caucasica\u003c/em\u003e leaf extracts, (ethanol extract, n-hexane fraction, and chloroform fraction). GC-MS analysis identified various bioactive compounds, including fatty acids, alcohols, and aromatic compounds, with known antioxidant, anti-inflammatory, and antimicrobial properties. The antioxidant activity, assessed via DPPH and ABTS assays, was highest in the chloroform fraction. The extracts also exhibited significant anti-cholinesterase and ant-diabetic potential, with CHFCC showing the strongest inhibition against α-glucosidase and α-amylase. In anti-inflammatory assays, CHFCC determined the highest inhibition of COX-2, while NHFCC showed the most potent 5-LOX inhibition. Future research should focus on in vivo validation of these effects, further investigation of molecular mechanism, formulation development for better bioavailability, toxicity and safety profiling, and exploring the synergistic effect of \u003cem\u003eC. caucasica\u003c/em\u003e with other medicinal compounds.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics Approval and Consent to Participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePlant specimens of \u003cem\u003eC. caucasica\u003c/em\u003e were obtained from Islamia College Peshawar, Pakistan. The samples were then transferred to the Department of Botany for taxonomic identification. Dr. Naveed Akhtar, Associate Professor, Department of Botany, Islamia College Peshawar, identified the plant species. A voucher specimen (ICP-BOT/102) was preserved in the departmental herbarium for future use.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions:\u003c/strong\u003e Conceptualization, Methodology, and Data curation, S.G., A.U.R., F.K., A.I., and R.J.; Experiments, A.U.R., M.S.J., S.N., and S.T.; Visualization and Writing Original Draft, A.U.R, A.M.M.A., and R.J.; supervision and arranging resources, R.J., A.I., and K.M.K.; \u0026nbsp;All authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments:\u003c/strong\u003e This work was carried out with the support of the \u0026ldquo;Cooperative Research Program for Agriculture Science and Technology Development (Project No. RS-2024-003222408)\u0026rdquo;, Rural Development Administration, Republic of Korea. The authors extend their appreciation to the Researcher Supporting Project number (RSPD2025R978), King Saud University, Riyadh, Saudi Arabia. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInstitutional Review Board Statement:\u003c/strong\u003e Not applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed Consent Statement:\u003c/strong\u003e Not Applicable.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability:\u0026nbsp;\u003c/strong\u003eData is provided within the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe author declared that there is no conflict of interest associated with this work.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eMittal, M., Gupta, N., Parashar, P., Mehra, V. \u0026amp; Khatri, M. Phytochemical evaluation and pharmacological activity of Syzygium aromaticum: a comprehensive review. \u003cem\u003eInt. J. Pharm. Pharm. Sci.\u003c/em\u003e \u003cb\u003e6\u003c/b\u003e, 67\u0026ndash;72 (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMujeeb, F., Bajpai, P. \u0026amp; Pathak, N. Phytochemical evaluation, antimicrobial activity, and determination of bioactive components from leaves of Aegle marmelos. \u003cem\u003eBioMed research international\u003c/em\u003e 497606 (2014). (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRoglic, G. WHO Global report on diabetes: A summary. \u003cem\u003eInt. J. Noncommunicable Dis.\u003c/em\u003e \u003cb\u003e1\u003c/b\u003e, 3\u0026ndash;8 (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChew, N. W. et al. The global burden of metabolic disease: Data from 2000 to 2019. \u003cem\u003eCell Metabol.\u003c/em\u003e \u003cb\u003e35\u003c/b\u003e, 414\u0026ndash;428 (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, J. et al. The protective effect of theaflavins on the kidney of mice with type II diabetes mellitus. \u003cem\u003eNutrients\u003c/em\u003e \u003cb\u003e15\u003c/b\u003e, 201 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRath, P. et al. A critical review on role of available synthetic drugs and phytochemicals in insulin resistance treatment by targeting PTP1B. \u003cem\u003eAppl. Biochem. Biotechnol.\u003c/em\u003e \u003cb\u003e194\u003c/b\u003e, 4683\u0026ndash;4701 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFeingold, K. R. Oral and injectable (non-insulin) pharmacological agents for the treatment of type 2 diabetes. \u003cem\u003eEndotext [Internet]\u003c/em\u003e (2024).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSharma, R. et al. Role of Shankhpushpi (Convolvulus pluricaulis) in neurological disorders: An umbrella review covering evidence from ethnopharmacology to clinical studies. \u003cem\u003eNeurosci. Biobehavioral Reviews\u003c/em\u003e. \u003cb\u003e140\u003c/b\u003e, 104795 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDeFronzo, R. A. et al. Type 2 diabetes mellitus. \u003cem\u003eNat. reviews Disease primers\u003c/em\u003e. \u003cb\u003e1\u003c/b\u003e, 1\u0026ndash;22 (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTran, N., Pham, B. \u0026amp; Le, L. Bioactive compounds in anti-diabetic plants: From herbal medicine to modern drug discovery. \u003cem\u003eBiology\u003c/em\u003e \u003cb\u003e9\u003c/b\u003e, 252 (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSharma, R. et al. Traditional Ayurvedic and herbal remedies for Alzheimer\u0026rsquo;s disease: from bench to bedside. \u003cem\u003eExpert Rev. Neurother.\u003c/em\u003e \u003cb\u003e19\u003c/b\u003e, 359\u0026ndash;374 (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen, Z. R., Huang, J. B., Yang, S. L. \u0026amp; Hong, F. F. Role of cholinergic signaling in Alzheimer\u0026rsquo;s disease. \u003cem\u003eMolecules\u003c/em\u003e \u003cb\u003e27\u003c/b\u003e, 1816 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eQureshi, T. \u0026amp; Chinnathambi, S. Histone deacetylase-6 modulates Tau function in Alzheimer's disease. \u003cem\u003eBiochim. et Biophys. Acta (BBA)-Molecular Cell. Res.\u003c/em\u003e \u003cb\u003e1869\u003c/b\u003e, 119275 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSharma, R., Kabra, A., Rao, M. \u0026amp; Prajapati, P. Herbal and holistic solutions for neurodegenerative and depressive disorders: leads from Ayurveda. \u003cem\u003eCurr. Pharm. Design\u003c/em\u003e. \u003cb\u003e24\u003c/b\u003e, 2597\u0026ndash;2608 (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSharifi-Rad, J. et al. Multi-target mechanisms of phytochemicals in Alzheimer\u0026rsquo;s disease: Effects on oxidative stress, neuroinflammation and protein aggregation. \u003cem\u003eJ. Personalized Med.\u003c/em\u003e \u003cb\u003e12\u003c/b\u003e, 1515 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar, A. \u0026amp; Singh, A. A review on Alzheimer's disease pathophysiology and its management: an update. \u003cem\u003ePharmacol. Rep.\u003c/em\u003e \u003cb\u003e67\u003c/b\u003e, 195\u0026ndash;203 (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHassan, H. A. et al. Isolation and characterization of novel acetylcholinesterase inhibitors from Ficus benghalensis L. leaves. \u003cem\u003eRSC Adv.\u003c/em\u003e \u003cb\u003e10\u003c/b\u003e, 36920\u0026ndash;36929 (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTaqui, R., Debnath, M., Ahmed, S. \u0026amp; Ghosh, A. Advances on plant extracts and phytocompounds with acetylcholinesterase inhibition activity for possible treatment of Alzheimer's disease. \u003cem\u003ePhytomedicine plus\u003c/em\u003e. \u003cb\u003e2\u003c/b\u003e, 100184 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eValarezo, E. et al. Enantiomeric composition, antioxidant capacity and anticholinesterase activity of essential oil from leaves of Chirimoya (Annona cherimola Mill). \u003cem\u003ePlants\u003c/em\u003e \u003cb\u003e11\u003c/b\u003e, 367 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCacoub, P. et al. Towards a common definition for the diagnosis of iron deficiency in chronic inflammatory diseases. \u003cem\u003eNutrients\u003c/em\u003e \u003cb\u003e14\u003c/b\u003e, 1039 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDoiphode, S., Lokhande, K. B., Ghosh, P., Swamy, K. \u0026amp; Nagar, S. Dual inhibition of cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) by resveratrol derivatives in cancer therapy: in silico approach. \u003cem\u003eJ. Biomol. Struct. Dynamics\u003c/em\u003e. \u003cb\u003e41\u003c/b\u003e, 8571\u0026ndash;8586 (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMeshram, M. A., Bhise, U. O., Makhal, P. N. \u0026amp; Kaki, V. R. Synthetically-tailored and nature-derived dual COX-2/5-LOX inhibitors: Structural aspects and SAR. \u003cem\u003eEur. J. Med. Chem.\u003c/em\u003e \u003cb\u003e225\u003c/b\u003e, 113804 (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEl-Miligy, M. M. et al. Towards safer anti-inflammatory therapy: synthesis of new thymol\u0026ndash;pyrazole hybrids as dual COX-2/5-LOX inhibitors. \u003cem\u003eJ. Enzyme Inhib. Med. Chem.\u003c/em\u003e \u003cb\u003e38\u003c/b\u003e, 294\u0026ndash;308 (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSL, M. Novel approach of multi-targeted thiazoles and thiazolidenes toward anti-inflammatory and anticancer therapy\u0026mdash;dual inhibition of COX-2 and 5-LOX enzymes. \u003cem\u003eMed. Chem. Res.\u003c/em\u003e \u003cb\u003e30\u003c/b\u003e, 236\u0026ndash;257 (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCantarello, E. et al. Human impacts on forest biodiversity in protected walnut-fruit forests in Kyrgyzstan. \u003cem\u003eJ. Sustainable Forestry\u003c/em\u003e. \u003cb\u003e33\u003c/b\u003e, 454\u0026ndash;481 (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAshafa, A., Sunmonu, T., Abass, A. \u0026amp; Ogbe, A. Laxative potential of the ethanolic leaf extract of Aloe vera (L.) Burm. f. in Wistar rats with loperamide-induced constipation. \u003cem\u003eJ. Nat. Pharmaceuticals\u003c/em\u003e. \u003cb\u003e2\u003c/b\u003e, 158\u0026ndash;162 (2011).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHartati, S., Angelina, M., Dewiyanti, I. \u0026amp; Meilawati, L. Isolation and characterization compounds from hexane and ethyl acetate fractions of Peperomia pellucida L. \u003cem\u003eJ. Trop. Life Sci.\u003c/em\u003e \u003cb\u003e5\u003c/b\u003e, 117\u0026ndash;122 (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEllman, G. L., Courtney, K. D., Andres Jr, V. \u0026amp; Featherstone, R. M. A new and rapid colorimetric determination of acetylcholinesterase activity. \u003cem\u003eBiochem. Pharmacol.\u003c/em\u003e \u003cb\u003e7\u003c/b\u003e, 88\u0026ndash;95 (1961).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJan, M. S. et al. Design, synthesis, in-vitro, in-vivo and in-silico studies of pyrrolidine-2, 5-dione derivatives as multitarget anti-inflammatory agents. \u003cem\u003eEur. J. Med. Chem.\u003c/em\u003e \u003cb\u003e186\u003c/b\u003e, 111863 (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBadshah, S. L. et al. Isolation, characterization, and medicinal potential of polysaccharides of Morchella esculenta. \u003cem\u003eMolecules\u003c/em\u003e \u003cb\u003e26\u003c/b\u003e, 1459 (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJan, M. S. et al. Synthesis of pyrrolidine-2, 5-dione based anti-inflammatory drug: in vitro COX-2, 5-LOX inhibition and in vivo anti-inflammatory studies. \u003cem\u003eLatin Am. J. Pharm.\u003c/em\u003e \u003cb\u003e38\u003c/b\u003e, 2287\u0026ndash;2294 (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSouilah, N. et al. Phenolic compounds from an algerian endemic species of Hypochaeris laevigata var. hipponensis and investigation of antioxidant activities. \u003cem\u003ePlants\u003c/em\u003e \u003cb\u003e9\u003c/b\u003e, 514 (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePena-Mart\u0026iacute;n, C., Serrano, R., Grindlay, G. \u0026amp; Crespo, M. B. Assessment of antioxidant capacity of Cystosphaera jacquinotii (Fucales, Ochrophyta) by two methods: DPPH and CUPRAC. (2024).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShah, P. \u0026amp; Modi, H. Comparative study of DPPH, ABTS and FRAP assays for determination of antioxidant activity. \u003cem\u003eInt. J. Res. Appl. Sci. Eng. Technol.\u003c/em\u003e \u003cb\u003e3\u003c/b\u003e, 636\u0026ndash;641 (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDome, P., Tombor, L., Lazary, J., Gonda, X. \u0026amp; Rihmer, Z. Natural health products, dietary minerals and over-the-counter medications as add-on therapies to antidepressants in the treatment of major depressive disorder: a review. \u003cem\u003eBrain Res. Bull.\u003c/em\u003e \u003cb\u003e146\u003c/b\u003e, 51\u0026ndash;78 (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLayek, B. \u0026amp; Mandal, S. Natural polysaccharides for controlled delivery of oral therapeutics: a recent update. \u003cem\u003eCarbohydr. Polym.\u003c/em\u003e \u003cb\u003e230\u003c/b\u003e, 115617 (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFadilah, N. I. M., Isa, I. L. M., Zaman, W. S. W. K., Tabata, Y. \u0026amp; Fauzi, M. B. The effect of nanoparticle-incorporated natural-based biomaterials towards cells on activated pathways: A systematic review. \u003cem\u003ePolymers\u003c/em\u003e \u003cb\u003e14\u003c/b\u003e, 476 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIqbal, A. et al. Phytochemistry, Laxative, Prokinetic and Spasmolytic Verification of Withania somnifera (L.) Dunal Using In-Vitro, In-Vivo and In-Silico Approaches. \u003cem\u003eJ. Biol. Regul. Homeost. Agents\u003c/em\u003e. \u003cb\u003e38\u003c/b\u003e, 3057\u0026ndash;3072 (2024).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBlickle, J., Andres, E. \u0026amp; Brogard, J. Current status of the treatment of type 2 diabetes mellitus. Alpha-glucosidase inhibitors. \u003cem\u003eLa. Revue de Med. Interne\u003c/em\u003e. \u003cb\u003e20\u003c/b\u003e, 379s\u0026ndash;383s (1999).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAbadan, Ş. et al. Synthesis and molecular modeling studies of naphthazarin derivatives as novel selective inhibitors of α-glucosidase and α-amylase. \u003cem\u003eJ. Mol. Struct.\u003c/em\u003e \u003cb\u003e1278\u003c/b\u003e, 134954 (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDirir, A. M., Daou, M., Yousef, A. F. \u0026amp; Yousef, L. F. A review of alpha-glucosidase inhibitors from plants as potential candidates for the treatment of type-2 diabetes. \u003cem\u003ePhytochem. Rev.\u003c/em\u003e \u003cb\u003e21\u003c/b\u003e, 1049\u0026ndash;1079 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDaniel, R., Mathew, B., Devi, K. \u0026amp; Augusti, K. Antioxidant effect of two flavonoids from the bark of Ficus bengalensis Linn in hyperlipidemic rats. \u003cem\u003eIndian J. Exp. Biol.\u003c/em\u003e \u003cb\u003e36\u003c/b\u003e, 902\u0026ndash;906 (1998).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMukhopadhyay, N., Shukla, A., Makhal, P. N. \u0026amp; Kaki, V. R. Natural product-driven dual COX-LOX inhibitors: Overview of recent studies on the development of novel anti-inflammatory agents. \u003cem\u003eHeliyon\u003c/em\u003e \u003cb\u003e9\u003c/b\u003e (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMd Idris, M. H. et al. Flavonoids as dual inhibitors of cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX): molecular docking and in vitro studies. \u003cem\u003eBeni-Suef Univ. J. Basic. Appl. Sci.\u003c/em\u003e \u003cb\u003e11\u003c/b\u003e, 117 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRudrapal, M. et al. Dual synergistic inhibition of COX and LOX by potential chemicals from Indian daily spices investigated through detailed computational studies. \u003cem\u003eSci. Rep.\u003c/em\u003e \u003cb\u003e13\u003c/b\u003e, 8656 (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMukherjee, P. K., Kumar, V., Mal, M. \u0026amp; Houghton, P. J. Acetylcholinesterase inhibitors from plants. \u003cem\u003ePhytomedicine\u003c/em\u003e \u003cb\u003e14\u003c/b\u003e, 289\u0026ndash;300 (2007).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePohanka, M. Inhibitors of acetylcholinesterase and butyrylcholinesterase meet immunity. \u003cem\u003eInt. J. Mol. Sci.\u003c/em\u003e \u003cb\u003e15\u003c/b\u003e, 9809\u0026ndash;9825 (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMathew, M. \u0026amp; Subramanian, S. In vitro screening for anti-cholinesterase and antioxidant activity of methanolic extracts of ayurvedic medicinal plants used for cognitive disorders. \u003cem\u003ePloS one\u003c/em\u003e. \u003cb\u003e9\u003c/b\u003e, e86804 (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTuzimski, T., Petruczynik, A., Szultka-Młyńska, M., Sugajski, M. \u0026amp; Buszewski, B. Isoquinoline alkaloid contents in Macleaya cordata extracts and their acetylcholinesterase and butyrylcholinesterase inhibition. \u003cem\u003eMolecules\u003c/em\u003e \u003cb\u003e27\u003c/b\u003e, 3606 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGreig, N. H. et al. Selective butyrylcholinesterase inhibition elevates brain acetylcholine, augments learning and lowers Alzheimer β-amyloid peptide in rodent. \u003cem\u003eProceedings of the National Academy of Sciences\u003c/em\u003e 102, 17213\u0026ndash;17218 (2005).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAyaz, M. et al. Anti-Alzheimer\u0026rsquo;s studies on β-sitosterol isolated from Polygonum hydropiper L. \u003cem\u003eFront. Pharmacol.\u003c/em\u003e \u003cb\u003e8\u003c/b\u003e, 697 (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTawfeek, N. et al. Cupressus arizonica greene: phytochemical profile and cosmeceutical and dermatological properties of its leaf extracts. \u003cem\u003eMolecules\u003c/em\u003e \u003cb\u003e28\u003c/b\u003e, 1036 (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAnokwuru, C., Anyasor, G., Ajibaye, O., Fakoya, O. \u0026amp; Okebugwu, P. Effect of extraction solvents on phenolic, flavonoid and antioxidant activities of three nigerian medicinal plants. \u003cem\u003eNat. Sci.\u003c/em\u003e \u003cb\u003e9\u003c/b\u003e, 53\u0026ndash;61 (2011).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLagouri, V., Bantouna, A. \u0026amp; Stathopoulos, P. A comparison of the antioxidant activity and phenolic content of nonpolar and polar extracts obtained from four endemic lamiaceae species grown in Greece. \u003cem\u003eJ. Food Process. Preserv.\u003c/em\u003e \u003cb\u003e34\u003c/b\u003e, 872\u0026ndash;886 (2010).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDe Sousa, D. B. et al. Metabolomic Profile of Volatile Organic Compounds from Leaves of Cashew Clones by HS-SPME/GC-MS for the Identification of Candidates for Anthracnose Resistance Markers. \u003cem\u003eJ. Chem. Ecol.\u003c/em\u003e \u003cb\u003e49\u003c/b\u003e, 87\u0026ndash;102 (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSudha, T., Chidambarampillai, S. \u0026amp; Mohan, V. GC-MS analysis of bioactive components of aerial parts of Fluggea leucopyrus Willd.(Euphorbiaceae). \u003cem\u003eJ. Appl. Pharm. Sci.\u003c/em\u003e \u003cb\u003e3\u003c/b\u003e, 126\u0026ndash;130 (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChirumamilla, P., Dharavath, S. B. \u0026amp; Taduri, S. GC\u0026ndash;MS profiling and antibacterial activity of Solanum khasianum leaf and root extracts. \u003cem\u003eBull. Natl. Res. Centre\u003c/em\u003e. \u003cb\u003e46\u003c/b\u003e, 127 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhaneian, M. T., Ehrampoush, M. H., Jebali, A., Hekmatimoghaddam, S. \u0026amp; Mahmoudi, M. Antimicrobial activity, toxicity and stability of phytol as a novel surface disinfectant. \u003cem\u003eEnviron. Health Eng. Manage. J.\u003c/em\u003e \u003cb\u003e2\u003c/b\u003e, 13\u0026ndash;16 (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh, B. R. in \u003cem\u003eProceedings of International conference and 28th Annual convention of IAVMI-2014 on Challenges and opportunities in animal health at the face of globalization and climate change, Department of Veterinary Microbiology and Immunology, DUVASU, Mathura, India.\u003c/em\u003e 26\u0026ndash;29.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYasin, Z. N. M. et al. Biological Activities and GCMS Analysis of the Methanolic Extract of Christia vespertilionis (LF) Bakh. F. Leaves. \u003cem\u003eAsian J. Med. Biomed.\u003c/em\u003e \u003cb\u003e4\u003c/b\u003e, 78\u0026ndash;88 (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNallathambi, A. \u0026amp; Bhargavan, R. GC/MS analysis of bioactive compounds in aqueous extract of cynodon dactylon. \u003cem\u003eIndian J. Public. Health Res. Dev.\u003c/em\u003e \u003cb\u003e10\u003c/b\u003e, 55\u0026ndash;59 (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNaveed, M. et al. Characterization and evaluation of the antioxidant, antidiabetic, anti-inflammatory, and cytotoxic activities of silver nanoparticles synthesized using Brachychiton populneus leaf extract. \u003cem\u003eProcesses\u003c/em\u003e \u003cb\u003e10\u003c/b\u003e, 1521 (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZlatić, N., Stanković, M. \u0026amp; Anticholinesterase Antidiabetic and anti-inflammatory activity of secondary metabolites of Teucrium species. \u003cem\u003eTeucrium Species: Biology Appl.\u003c/em\u003e, 391\u0026ndash;411 (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZeb, A. \u0026amp; Zeb, A. Molecular Mechanism of Phenolic Antioxidants. \u003cem\u003ePhenolic Antioxid. Foods: Chem. Biochem. Anal.\u003c/em\u003e, 413\u0026ndash;434 (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKravchenko, I., Kireva, M. \u0026amp; Alekseeva, E. Synthesis and anti-inflammatory activity of ibuprofen esters. \u003cem\u003ePharm. Chem. J.\u003c/em\u003e \u003cb\u003e48\u003c/b\u003e, 313\u0026ndash;316 (2014).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Supplementary Material","content":"\u003cp\u003eSupplementary Material are not available with this version.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Celtus caucasica, GC-MS, Antioxidant, Antidiabetic, Anticholinesterase, and Anti-inflammatory Properties","lastPublishedDoi":"10.21203/rs.3.rs-6223687/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6223687/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study evaluates the phytochemical composition and biological activities of the ethanol extract of \u003cem\u003eCeltis caucasica\u003c/em\u003e leaves. Gas Chromatography-Mass Spectrometry (GC-MS) identified sixteen bioactive compounds in the ethanol extract of \u003cem\u003eC. caucasica\u003c/em\u003e (ETCC), including 4-O-Methylmannose, Guaifenesin, Hexadecanoic acid derivatives, and Phytol, while the chloroform fraction of \u003cem\u003eC. caucasica\u003c/em\u003e (CHFCC) contained twenty compounds, notably Phthalic acid di(2-propylpentyl) ester (11.94%) and Octadec-9-enoic acid (11.76%), known for their antioxidant, anti-inflammatory, and neuroprotective properties. Fourier-transform infrared (FT-IR) spectroscopy revealed diverse functional groups in ETCC and strong aromatic peaks with metal-organic complexes in CHFCC. Biological evaluations showed CHFCC had the highest acetylcholinesterase (AChE, 86.44%, IC₅₀ = 13.2 \u0026micro;g/mL) and butyrylcholinesterase (BChE, 92.67%, IC₅₀ = 9.66 \u0026micro;g/mL) inhibition, as well as potent α-glucosidase (88.61%, IC₅₀ = 11.99 \u0026micro;g/mL) and α-amylase inhibition (91.36%, IC₅₀ = 4.22 \u0026micro;g/mL), indicating strong antidiabetic potential. CHFCC also exhibited the highest COX-2 inhibition (88.61%), while the n-hexane fraction (NHFCC) showed the strongest 5-LOX inhibition (88.88%). Antioxidant assays revealed CHFCC had the highest radical scavenging activity (IC₅₀ = 77.24 \u0026micro;g/mL for DPPH and 60.67 \u0026micro;g/mL for ABTS), though lower than ascorbic acid (IC₅₀ = 2.81 \u0026micro;g/mL and 4.6 \u0026micro;g/mL, respectively). These findings highlight \u003cem\u003eC. caucasica\u003c/em\u003e as a promising source of bioactive compounds with therapeutic potential.\u003c/p\u003e","manuscriptTitle":"Phytochemical Profiling and Bioactivity of Celtis caucasica: Antioxidant, Antidiabetic, Anticholinesterase, and Anti-inflammatory Potential","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-28 17:24:24","doi":"10.21203/rs.3.rs-6223687/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"9229c9fc-bc57-467e-8c07-6bb6cf553d3e","owner":[],"postedDate":"March 28th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":46109074,"name":"Biological sciences/Biochemistry"},{"id":46109075,"name":"Biological sciences/Chemical biology"},{"id":46109076,"name":"Biological sciences/Microbiology"},{"id":46109077,"name":"Biological sciences/Plant sciences"},{"id":46109078,"name":"Earth and environmental sciences/Biogeochemistry"}],"tags":[],"updatedAt":"2025-06-02T09:38:28+00:00","versionOfRecord":[],"versionCreatedAt":"2025-03-28 17:24:24","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6223687","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6223687","identity":"rs-6223687","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}