Peanut seeds were meticulously hand-harvested from the El Oued region in Algeria. The seeds were dried at room temperature for 1 month and then conserved at 4°C until their use. About 1 g of dried seeds was ground into a coarse powder to optimize the surface area for efficient extraction. This powder was then subjected to Soxhlet extraction (FOSS Soxtec™ 8,000) using n-hexane (≥98% purity, Sigma-Aldrich) as the solvent, chosen for its efficacy in oil extraction. The extraction process spanned 6 h, ensuring thorough oil recovery. For each 1 g of peanut kernels placed in the thimbles, 25 mL of n-hexane was poured into the extraction vessels. To maintain the oil’s integrity and prevent degradation, it was stored in amber glass bottles at a controlled temperature of 4°C, minimizing light exposure until further analysis.

The lipophilic extract obtained from peanut underwent a methylation process prior to GC-MS analysis, as the fatty acids in their original form are not sufficiently volatile for direct gas chromatographic injection. The methylation step converts the fatty acids into fatty acid methyl esters (FAMEs), which are more suitable for gas chromatography due to their increased volatility. Briefly, the extracted peanut oil (1 mL) was subjected to a methylation process to convert the fatty acids into their corresponding fatty acid methyl esters (FAME). Since the oil primarily contains triacylglycerols, which are not directly analyzable by GC-MS, the lipids were first reacted with 2 mL of methanol and 200 µL of concentrated sulfuric acid (98%) as a catalyst. This mixture was heated at 70°C for 2 h to complete the esterification reaction. After cooling, the FAME were extracted by adding 2 mL of hexane and 1 mL of distilled water. The organic phase (hexane) containing the FAME was separated and washed with a saturated sodium chloride solution (1 mL) to remove any remaining water-soluble impurities. The FAME were then dried over anhydrous sodium sulfate (0.2 g) and filtered before being concentrated to 1 mL under a stream of nitrogen gas.

GC-MS analysis was performed with a Hewlett-Packard 6,890 gas chromatograph coupled with a Hewlett-Packard 5,973 mass spectrometer. The GC system featured a capillary column (60 m × 0.32 mm inner diameter, 0.25 µm film thickness) and a flame ionization detector (FID). Helium served as the carrier gas at a flow rate of 1 mL/min. The temperature program started at 60°C, increased to 250°C at a rate of 3°C/min, and was held at 250°C for 10 min. The detector and injector temperatures were set at 245°C and 250°C, respectively. The mass spectrometer was operated in electron ionization mode with an ionization potential of 70 eV. Compound identification was achieved by comparing retention times and mass spectra with NIST and Wiley libraries, and matching fragmentation patterns and retention indices which were reported in the literature.

The inhibition of Butyrylcholinesterase (BChE) is vital for assessing the potential of peanut oil constituents to impede the breakdown of acetylcholine, a neurotransmitter closely associated with the progression of Alzheimer’s disease. BChE inhibition was evaluated using (Ellman et al., 1961). In a 96-well microplate, 20 µL of BChE from equine serum (≥10 units/mg protein, E.C. 3.1.1.8, Sigma-Aldrich) solution (6.85 × 10⁻³ U) was mixed with 150 µL of sodium phosphate buffer (pH 8.0) and 10 µL of peanut oil at various concentrations. After a 15-min incubation at 25°C, 10 µL of DTNB [5,5′-dithio-bis(2-nitrobenzoic) acid] (≥98%, BioReagent, Sigma-Aldrich) (0.5 mM) and 10 µL of butyrylcholine iodide (≥98%, Sigma-Aldrich) (0.2 mM) were added. Absorbance was measured at 412 nm initially and again after 15 min using a microplate reader. Galantamine, a known BChE inhibitor, was used as a positive control and treated in the same manner as the peanut oil extracts. The percentage of BChE enzyme inhibition was calculated using the following formula: Inhibition  % = Abs Control   –   Abs Sample / Abs Control * 100 Abs Control : BChE enzyme − extract ;   Abs Sample : BChE enzyme + oil

Statistical analyses were conducted to evaluate the significance of the experimental results. Linear regression analysis was applied to determine the half-maximal inhibitory concentration (IC₅₀) and A₀.₅ values, which represent the concentration of peanut oil or reference compounds required to achieve 50% inhibition or antioxidant activity, respectively. To identify significant differences between groups, one-way analysis of variance (ANOVA) was performed, with a significance threshold set at p < 0.05. When significant differences were detected, the Tukey post hoc test was used for multiple comparisons. All statistical analyses were performed using XLSTAT software. Results are presented as the mean ± standard deviation (SD) of three independent measurements.

Canonical SMILES for the identified compounds, including Oleic acid, Palmitic acid, Stearic acid, and Oxiraneoctanoic acid, 3-octyl, were retrieved from the PubChem database (NCBI, 2023). Physicochemical properties were predicted using Lipinski’s rule of five on the Swiss ADME platform (SwissADME, 2023), assessing parameters such as size, polarity, lipophilicity, solubility, flexibility, and unsaturation. ADMET properties were analyzed using the admetSAR 2.0 platform (IMMD, 2023), focusing on human intestinal absorption (HIA), Caco-2 permeability, blood-brain barrier (BBB) permeability, and human bioavailability (HOB). Cytotoxic and anticancer effects of the compounds on tumor cells were evaluated using the CLC-Pred server (Way2drug, 2023). The compounds extracted from peanut oil were evaluated for their anticancer activity against a variety of cancer cell lines, including melanoma A2058, metastatic melanoma SK-MEL-1, non-small cell lung cancer NCI-H838, lung carcinoma DMS-114, ovarian adenocarcinoma IGROV, and leukemia H9. These assays aimed to assess the cytotoxic potential of the oil’s major constituents, with each compound demonstrating varying degrees of effectiveness across the different cancer type. This prediction evaluates the probability of activity (Pa) versus inactivity (Pi), with a higher Pa score indicating a greater likelihood of activity compared to inactivity, though not guaranteeing it. This combined approach aims to identify promising compounds for further experimental validation, facilitating the evaluation of their therapeutic potential in cancer treatment.

Molecular docking analysis was performed on key compounds identified in peanut oil to evaluate their binding affinities with Butyrylcholinesterase (BChE), aiming to assess their potential anti-Alzheimer’s activity. The 3D structure of BChE (PDB ID: 2XQF) was obtained from the Protein Data Bank and processed for docking using LeadIT 2.1.8 (Biosolveit, 2023). All heteroatoms, cofactors, and non-essential water molecules were removed from the crystallized structure, and any missing atoms were added. The active site was defined by residues within a 6.5 Å radius around the inhibitor, and additional residues critical for maintaining the binding cavity were included. The protonation states and side-chain orientations were optimized. Ligand structures were prepared using LigPrep in Schrödinger’s Maestro 11.3 (Schrodinger Release, 2015-1, 2015), considering all possible tautomers and protonation states. Molecular docking was carried out using FlexX 2.1.8 (Rarey et al., 1996), with binding energies (ΔG in kJ/mol) indicating the strength of interactions. The co-crystallized ligand was redocked to validate docking parameters, ensuring an RMSD value of less than 2 Å.

Plants have long been central to medicinal research, with peanut oil providing valuable insights into its composition and health benefits. Despite its traditional use, studies on peanut oil remain relatively limited (Djeghim et al., 2021; Djeghim et al., 2022). This study highlights that the hexane extraction method is particularly effective for obtaining higher yields of bioactive compounds. Hexane excels at extracting non-polar substances like lipids and specific phytochemicals due to its low viscosity and high volatility (Bhuiya et al., 2020). Improved yields with hexane extraction depend on factors such as solvent-to-solute ratio and extraction techniques, allowing for the capture of a broader range of bioactive compounds essential for unlocking potential health benefits (Shad et al., 2012; Zio et al., 2021). The oil extracted from peanuts exhibited a high yield of 51.85% ± 1.76%, indicating an efficient process that produces a substantial amount of oil from the seed material. The extracted oil was noted for its uncolored appearance and pleasant odor, attributed to the presence of methyl nonyl ketone. Analysis of the oil revealed noteworthy levels of bioactive compounds, with a total phenolic content measured at 28.21 ± 2.43 µg GAE/mg, indicating a rich presence of phenolic constituents known for their antioxidant properties. Similarly, the total flavonoid content was found to be 27.72 ± 1.25 µg QE/mg, underscoring the oil’s potential health benefits given the antioxidant and anti-inflammatory effects of flavonoids (Table 1).

GC-MS analysis of peanut oil provided a comprehensive fatty acid profile, accounting for 99.9% of its composition (Figure 1). The predominant fatty acid identified was oleic acid (41.98%), followed by palmitic acid (26.90%) and stearic acid (8.11%), which is consistent with prior research (Nile and Park, 2013). This detailed breakdown illustrates the balance of various fatty acids, categorized into saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA), each contributing to the oil’s nutritional and functional properties (Table 2). To ensure the accuracy and reliability of GC-MS analysis, the retention indices of the identified compounds were recalculated using the Van Den Dool-Kratz equation (Zhokhov et al., 2018). This involved applying the retention times of standard n-alkanes to derive the indices for each compound. Subsequently, these recalculated indices were compared with those reported in the NIST and Adams literature databases.

(1) Major Compounds: The main fatty acid in peanut oil is oleic acid, which makes up 41.98% of its total fatty acid content. This monounsaturated fatty acid is recognized for its heart health benefits, such as lowering LDL cholesterol and raising HDL cholesterol. This makes it important for promoting cardiovascular health, reducing inflammation, and helping to maintain cell membrane fluidity and improve insulin sensitivity (Lopez-Huertas, 2010; Yin et al., 2007; Zhao et al., 2019; Anavi-Cohen et al., 2023). The second most common fatty acid is palmitic acid, which comprises 26.90% of the oil. While palmitic acid is a saturated fatty acid linked to raising LDL cholesterol levels, its effects can be lessened by the presence of oleic acid and other beneficial compounds in peanut oil (Astrup et al., 2011; Bhuiya et al., 2020; Shad et al., 2012). Stearic acid accounts for 8.11% of the oil and is another important saturated fatty acid. Unlike palmitic acid, stearic acid does not negatively affect LDL cholesterol levels and may even help lower them, thus supporting heart health (Tong et al., 2020; Slavin, 2004). Stearic acid’s neutral effect on cholesterol makes it a promising area for further research into its benefits for lipid profiles.

The antioxidant potential of the extracted peanut oil was assessed using the DPPH radical scavenging assay and the CUPRAC assay. As shown in Table 3, peanut oil exhibited relatively weak antioxidant activity in both tests, with an IC₅₀ value of 1715.92 ± 93.28 μg/mL for the DPPH assay and an A_0.5 value of 265.96 μg/mL for the CUPRAC assay. These values are significantly higher (p < 0.001) than those of the reference compounds BHA (IC₅₀: 15.68 ± 0.25 μg/mL; A_0.5: 1.79 ± 0.07 μg/mL) and BHT (IC₅₀: 6.68 ± 0.06 μg/mL; A_0.5: 4.58 ± 0.10 μg/mL), indicating a much lower antioxidant potential for peanut oil. This indicates that, despite the presence of phenolic and flavonoid compounds, their concentrations or types may not effectively neutralize free radicals or reduce metal ions. Additionally, peanut oil has lower antioxidant activity compared to oils like sesame and olive oil (Prabasheela et al., 2015). The higher IC₅₀ value in the DPPH assay suggests reduced radical scavenging ability, likely due to the loss of hydrophilic antioxidants during extraction. While peanut oil contains polyphenolic compounds, their effectiveness is diminished by non-polar lipids, unlike oils rich in polyunsaturated fatty acids, which typically show higher antioxidant activity due to compounds like tocopherols (Kapoor et al., 2021; Phuah et al., 2022). Moreover, interactions between tocopherols and phenolic acids may further lessen antioxidant activity (Palafox-Carlos et al., 2012).

Anti-Alzheimer’s activity is often linked to the inhibition of Butyrylcholinesterase (BChE), an enzyme responsible for breaking down acetylcholine, a neurotransmitter crucial for memory and cognitive function. In Alzheimer’s disease, reduced acetylcholine levels contribute to cognitive decline, making BChE inhibition a potential therapeutic strategy to help maintain acetylcholine levels in the brain. The inhibitory effect of the peanut oil on BChE was measured and compared to the standard inhibitor, Galantamine. The data, presented in Table 4, show that the peanut oil demonstrated a weak BChE inhibitory capacity. At a concentration of 200 μg/mL, the oil achieved an inhibition percentage of 36.47% ± 3.71% which was significantly lower compared to Galantamine 96.98 ± 0.94 μg/mL, as indicated by ** (p = 0.00028), at the same concentration and had an IC₅₀ value of 11.61 ± 0.22 μg/mL. The observed inhibition by peanut oil, although limited, is still relevant to its potential anti-Alzheimer’s activity, as even partial inhibition of BChE could help sustain acetylcholine levels. To date, there have been no reports in the literature on the BChE inhibitory capacity of Algerian peanut oil, making this a novel finding. These results are in line with previous studies, such as one by (Bozorgi et al., 2021), which showed that the aqueous extract of areca nuts exhibited strong anti-Alzheimer capacity through inhibition of amyloid β aggregation. Similarly, (Chauhan and Chauhan, 2020), highlighted the protective effects of walnut-enriched diets against AD.

The physicochemical and ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) profiles of compounds found in peanut oil, along with their cytotoxic effects, are critical for evaluating the therapeutic potential of these compounds, particularly in targeting various cancer cell lines. The analysis combines cytotoxicity data with the physicochemical characteristics of the compounds, offering a comprehensive overview of their anticancer activities.

Table 5 summarizes the properties of the molecules identified in peanut oil, detailing their molecular weight, chemical formula, structure, and canonical SMILES. All compounds comply with Lipinski’s Rule of Five, suggesting good oral bioavailability. Notably, oxiraneoctanoic acid, 3-octyl, with a log p-value below 5, demonstrates favorable lipophilicity, highlighting its potential as a drug-like candidate. This thorough analysis emphasizes the fundamental characteristics of these compounds, with a special focus on oxiraneoctanoic acid, 3-octyl, for its promising prospects in drug development.

Table 6 presents the ADMET profiles of the major compounds in peanut oil, offering insights into their pharmacokinetic and toxicological characteristics. Each compound exhibited excellent absorption capabilities, effectively crossing the human intestinal barrier (HIA+), Caco-2 cell layer, and blood-brain barrier (BBB). All four compounds showed positive HIA+ and Caco-2 permeability, indicating strong potential for effective oral administration and systemic bioavailability. Importantly, none of the compounds acted as substrates or inhibitors of P-glycoproteins, which play a significant role in drug resistance.

The ADMET analysis indicates that oleic acid localizes in the plasma membrane, while palmitic acid, stearic acid, and oxiraneoctanoic acid, 3-octyl, localize in the mitochondria, which may influence their biological effects. Most compounds inhibited CYP1A2, except for oxiraneoctanoic acid, 3-octyl, indicating significant hepatic metabolism for the majority. None inhibited CYP3A4 or CYP2D6, which reduces the risk of drug-drug interactions. Toxicity studies revealed that oleic, palmitic, and stearic acids can cause eye corrosion, skin irritation, and mitochondrial toxicity, while oxiraneoctanoic acid, 3-octyl, showed no such effects. Hepatotoxicity was noted for palmitic, stearic, and oxiraneoctanoic acids, while nephrotoxicity was associated with oxiraneoctanoic acid, 3-octyl. Plasma protein binding was moderate for all compounds, with oxiraneoctanoic acid, 3-octyl, demonstrating the highest binding affinity. Oleic, palmitic, and stearic acids were classified as low toxicity (Class IV), while oxiraneoctanoic acid, 3-octyl, was classified as moderate toxicity (Class III).

Extracting bioactive compounds or fractions is a critical first step in traditional medicinal plant research, followed by qualitative and quantitative analyses to identify the active components (Ceylan et al., 2015). However, this process is often time-consuming and costly (Bucao and Solidum, 2021). To improve research efficiency, modifications to traditional approaches have been explored. As computer technologies advance, in silico methods such as the simulation of compound-target interactions have become increasingly reliable and accurate (Ntie-Kang et al., 2013). These computational approaches allow for the rapid identification of potential relationships between compounds and biological targets, facilitating faster drug discovery. One such advancement is network pharmacology, a technology that enables researchers to explore complex interactions between multiple compounds and their diverse activity targets. For this reason, the objective of this study was shaped by the promise of network pharmacology, aiming to apply these modern tools to streamline the discovery process and understand the multifaceted interactions of bioactive compounds in peanut oil.

The cytotoxicity data in Table 7 highlight the strong antitumor potential of the primary fatty acids in peanut oil, including oleic acid, palmitic acid, stearic acid, and oxiraneoctanoic acid, 3-octyl. Oleic acid exhibited significant cytotoxic effects across various cancer cell lines, especially against melanoma (A2058 and SK-MEL-1) and non-small cell lung cancer (NCI-H838), indicating broad-spectrum antitumor activity. Palmitic acid also showed notable cytotoxicity, particularly against lung carcinoma and metastatic melanoma. Stearic acid mirrored the cytotoxic profile of palmitic acid, reinforcing its potential against melanoma and lung cancers.

Oxiraneoctanoic acid, 3-octyl, distinguished itself with potent activity against leukemia (H9), demonstrating specific effects on hematopoietic and lymphoid tumors, as well as significant activity against lung carcinoma. This specificity positions it as a promising candidate for targeted anticancer therapies. The combined ADMET, physicochemical, and cytotoxicity profiles suggest that these fatty acids, particularly oleic, palmitic, and stearic acids, could be valuable in treating various cancer types, while oxiraneoctanoic acid, 3-octyl, stands out for its specific potency against leukemia. This finding aligns with (Hammad et al., 2023), who reported that peanut (A. hypogaea L.) skin ultrasound extract exhibited significant inhibitory activity against cancer cell lines, including MCF-7, HepG-2, HCT-116, and PC-3. Additionally (Jeeunngoi et al., 2024), found that combining ethanolic peanut skin extract with either cisplatin or 5-fluorouracil had a pronounced anticancer effect on HeLa cells. Furthermore (Tsai et al., 2024), revealed that an ethyl acetate fraction of crude peanut skin extract, alongside a methanolic fraction, exhibited the highest cytotoxicity against colorectal cancer (CRC) and melanoma cells, while sparing nonmalignant human skin fibroblasts. These results support the potential of peanut oil and its constituents in cancer treatment strategies.

Table 8 outlines the findings from the docking study, assessing the physicochemical properties of key compounds from peanut oil using the Swiss ADME tool. Parameters such as molecular weight (MW), solubility (Log S), number of rotatable bonds, hydrogen bond acceptors and donors, topological polar surface area (TPSA), and consensus logP values were evaluated. Fatty acids like oleic, palmitic, and stearic acids exhibited moderate insolubility, with log S values indicating limited aqueous solubility. Their lipophilicity, reflected in consensus logP values between 5.2 and 5.93, suggests a tendency to favor lipid-rich environments, potentially influencing their bioavailability and metabolic pathways. Oxiraneoctanoic acid, 3-octyl, with a higher MW of 298.46 g/mol and a consensus logP of 4.88, presents a unique profile, characterized by a greater TPSA of 49.83 Å². This suggests different interaction dynamics within biological systems compared to the more lipophilic fatty acids, indicating a distinct mechanism that could influence its activity in biological contexts.

It is important to note that the findings presented are based on in silico predictions and require empirical validation through laboratory experiments to confirm their accuracy. However, several studies have demonstrated the potential anticancer effects of various phytochemicals found in plant-based oils (Hammad et al., 2023; Jeeunngoi et al., 2024; Tsai et al., 2024). These bioactive compounds, such as oleic acid, palmitic acid, and stearic acid, exhibit notable cytotoxic properties against a wide range of cancer cell lines. Despite these promising results, the exact molecular pathways through which these compounds exert their anticancer effects remain underexplored. Future research involving pathway analysis and gene expression profiling will be essential to fully elucidate the mechanisms of action of these compounds and establish their true therapeutic potential.

Molecular docking simulations of 20 primary compounds derived from peanut oil were conducted within the active site of Butyrylcholinesterase (BChE). These simulations revealed significant correlations between the experimental inhibitory activities and the docking results, as summarized in Table 9. BChE was selected for this study due to its crucial role in metabolizing acetylcholine, a neurotransmitter vital for memory and cognitive functions. In Alzheimer’s disease, acetylcholine levels are markedly reduced, contributing to cognitive decline. By inhibiting BChE, we can slow the degradation of acetylcholine, potentially aiding in maintaining its levels in the brain. Thus, targeting the BChE active site presents a promising strategy for managing neurodegenerative disorders like Alzheimer’s.

The docking studies focused on two compounds: Galantamine, used as a standard, and 6-Methyl Octahydro-Coumarin derived from peanut oil, with their binding interactions illustrated in Figure 2. These simulations evaluated how each compound interacts with BChE, shedding light on their enzyme-inhibitory potential and therapeutic relevance for neurodegenerative diseases. As shown in Figure 2, both compounds predominantly bind at the bottom of the catalytic anionic site (CAS) of BChE, a region crucial for their inhibitory function. The CAS, highlighted in blue, and the peripheral anionic site (PAS), marked in red, illustrate how these compounds engage with the enzyme’s active site. The ligands, visualized with carbon atoms in green, oxygen in red, and nitrogen in blue, are strategically positioned to occupy key regions within the CAS, reinforcing their strong inhibitory potential.

Galantamine, a well-known cholinesterase inhibitor, demonstrated a docking score of −23.4 kJ/mol, indicating strong inhibitory potency consistent with its high experimental BChE inhibitory activity. In contrast, 6-Methyl Octahydro-Coumarin had a docking score of −15.86 kJ/mol, marking it as the most promising BChE inhibitor among the constituents of peanut oil. This compound was further analyzed to elucidate its binding interactions within the enzyme’s active site. Both Galantamine and 6-Methyl Octahydro-Coumarin primarily interact at the bottom of the catalytic anionic site (CAS) of BChE, a critical region for enzyme inhibition. Despite its favorable docking score, 6-Methyl Octahydro-Coumarin exhibited slightly lower inhibitory activity than Galantamine, likely due to forming fewer hydrogen bonds within the active site. Galantamine established five hydrogen bonds with key residues (Ser198, Gly116, Gly117, and His438), whereas 6-Methyl Octahydro-Coumarin formed three hydrogen bonds (with Gly116, Gly117, and Ala199).

Other compounds displayed varying docking scores, indicating different binding affinities to BChE. For instance, Azelaaldehydic acid (−10.95 kJ/mol) and Nonanedioic acid (−11.36 kJ/mol) exhibited moderate binding potential, while fatty acids like Palmitic acid (−4.83 kJ/mol) and Oleic acid (−4.36 kJ/mol) showed lower scores, suggesting weaker interactions with BChE. These results highlight the significance of hydrogen bonding and specific binding interactions in determining inhibitory potency. As illustrated in Table 9 and Figure 2, these molecular docking studies provide valuable insights into the potential of peanut oil constituents as BChE inhibitors, with implications for treating Alzheimer’s and other neurodegenerative conditions.

Figure 3 displays the most likely binding orientations of each compound as determined by docking simulations. The catalytic anionic site (CAS) is shown in blue, while the peripheral anionic site (PAS) is in red. Ligand atoms are color-coded: carbon in green, oxygen in red, and nitrogen in blue. This positioning emphasizes key interactions within the active site that contribute to the inhibitory effects of the compounds. The difference in BChE inhibitory potency between Galantamine and 6-Methyl Octahydro-Coumarin stems from their distinct hydrogen bonding patterns. Galantamine forms five hydrogen bonds with residues, enhancing its stability and inhibitory efficacy in the BChE active site, while 6-Methyl Octahydro-Coumarin forms only three, which may explain its slightly lower inhibitory potency despite a favorable docking score. Additionally, the hydrophobic interactions, indicated in green, contribute to the binding stability and overall effectiveness of these compounds as BChE inhibitors. Specifically, 6-Methyl Octahydro-Coumarin forms three hydrogen bonds with Gly116, Gly117, and Ala199, whereas Galantamine establishes five hydrogen bonds with Ser198, Gly116, Gly117, and His438. This differential binding pattern underscores the importance of specific amino acid interactions in enhancing or limiting inhibitory efficacy (Gonzalez et al., 2016).

The low concentration of 6-Methyl Octahydro-Coumarin (1%) in the studied oil presents challenges for extraction and purification, necessitating optimization of traditional methods and a cost-effectiveness evaluation of the refining process. Although the oil is rich in polyunsaturated fatty acids (PUFAs) like oleic acid, known for their neuroprotective effects, the absence of strong BChE inhibitors likely accounts for the moderate effects observed. This aligns with findings by Lauritzen et al. (2000), which emphasize that while PUFAs can enhance cognitive health, they do not significantly inhibit cholinesterase directly. Therefore, while peanut oil may not be a potent BChE inhibitor, it could still provide neuroprotective benefits when included in the diet. Docking simulations reveal that 6-Methyl Octahydro-Coumarin has the highest binding affinity to the BChE catalytic site, yet its binding strength is much lower than that of established inhibitors like Galantamine. This suggests that while some minor components of the oil may interact with BChE, they lack the potency for significant therapeutic impact. Previous studies, such as those by (Song et al., 2019; Sakurai et al., 2021), indicate that the cognitive benefits of oleic acid may relate to its role in modulating gene expression associated with neuroinflammation and synaptic health rather than direct cholinesterase inhibition. Thus, exploring other constituents or combining peanut oil with stronger inhibitors may yield better health outcomes.