Effect of Oil Extraction Process on the Fatty Acid Composition of Groundnut Oil

Effect of Oil Extraction Process on the Fatty Acid Composition of Groundnut Oil | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Effect of Oil Extraction Process on the Fatty Acid Composition of Groundnut Oil Keotshepile Precious Bojang, Aparna Kuna This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7142039/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 10 You are reading this latest preprint version Abstract Groundnut ( Arachis hypogaea L.) is one of the important oilseed crops in the world, which is utilized for cooking and in confectionery, and is a rich source of valuable antioxidants like vitamin E and plant sterols especially β-sitosterol. In this study, we attempted to study the differences in fatty acid composition in commercially available refined groundnut oil (CRGNO), hexane extracted crude groundnut oil (HCGNO) and cold pressed groundnut oil (CPGNO).The ratio of unsaturated fatty acid to fatty acids was 5.47 ± 3.09, 5.91 ± 2.96, and 5.34 ± 3.11 for CPGNO, CRGNO and HCGNO respectively. Oleic acid (C18:1n9cis) and Linoleic acids (C18:2n6cis) were the major unsaturated fatty acidsand Palmitic acid (C16:0)was the highest saturated fatty acid identified in all the three GNO oils.The total unsaturated fatty acid content was high in all the three oils (86.23 ± 0.23% in CRGNO, 85.12 ± 0.26% in HCGNO and 84.74 ± 0.17% in CPGNO).A lower percentage of saturated fatty acids was observed in all the three oils i.e., 15.93 ± 5.95%, 15.48 ± 6.00% and 14.58 ± 5.68% in HCGNO, CPGNO and CRGNO respectively. From the fatty acid analysis of the three oils, it was observed that there was minor variation in the Oleic acid (C18:1n9cis) and Linoleic acids (C18:2n6cis) content and no significant difference in other fatty acids composition among the three samples. Cold pressed and hexane extracted crude GNO had higher essential fatty acid content (linoleic acid (C18:2n6cis) compared to refined GNO, indicating the effect of oil extraction on the fatty acid composition. Groundnut Cold-pressed Hexane extracted oil Refining Fatty acids Figures Figure 1 Figure 2 Figure 3 Figure 4 INTRODUCTION The legume Arachis hypogaea , commonly known as peanut or groundnut, is a very important food crop throughout the tropics and subtropics. Groundnut is one of the most widely used legumes for its nutrition and taste, and occupies a rank of major oilseed crop in the world, due to its role in health promoting effect. Groundnut oil (GNO) owing to its unique organoleptic properties associated with its cardioprotective and anti-inflammatory properties has found its place on the highly competitive international edible oil market (Shamim et al.,2014). Its versatile applications span culinary realms, ranging from direct consumption to the production of cooking oil and confectionery products. Beyond its gastronomic significance, groundnut is renowned for its rich composition of bioactive compounds, including tocopherols (vitamin E) and phytosterols, notably β-sitosterol, which confer antioxidant and cholesterol-lowering properties (Maestri, 2024 ;Shin et al.,2009). The primary economic value of groundnut lies in its oil content, which typically ranges from 40–50% of the kernel weight, making it a substantial source of vegetable oil worldwide (Janila, 2016). Edible oils are often subject to intentional or unintentional mixing, such as blending cold-pressed oils with solvent-extracted or refined variants, which is frequently done to reduce production costs or enhance yield. This practice raises concerns over authenticity and quality, particularly when higher value oils are replaced with cheaper alternatives (Bojang et al.,2021). For example, GNO is blended with ricebran oil, sunflower oil, soybean oil, palm oil, or canola oil in varying proportions, or adulterated by adding its flavor to cheaper oils to fake it (Jee,2009 ;Fang et al.,2013;Xie et al.,2013).The main compound in vegetable oil is triglyceride (> 95%), which is composed of fatty acids (Ai et al.,2009). Fatty acid profiles can serve as a unique characteristic feature of edible vegetable oil. Fatty acids composition of vegetable oils is formed by a mixture of saturated and unsaturated fatty acids classified according to the number of unsaturated bonds as monounsaturated (MUFAs) or polyunsaturated fatty acids (PUFAs). The fatty acid profile of groundnut oil (GNO) is a critical determinant of its nutritional quality and oxidative stability. Predominantly, GNO is characterized by a high proportion of unsaturated fatty acids, particularly oleic acid (C18:1n9cis) and linoleic acid (C18:2n6cis), which collectively constitute over 80% of the total fatty acids (Musalima et al.,2019).These unsaturated fatty acids are essential for human health, playing vital roles in cell membrane integrity, eicosanoid synthesis, and cholesterol metabolism (Simopoulos et al.,2016). To extract oil from seeds, fruits and other oil-bearing materials, different extraction methods are used, for example cold pressing, use of polar and non-polar solvents depending on the intended use (Nagaraj, 2009 ;Adebisi and Eriola,2019;Adebayo et al.,2015). The extraction method employed to obtain GNO significantly influences its final composition and quality. However, most of the time commercial mills employs a combination of various unit operations including conditioning, flaking and solvent extraction whereby the oil-bearing materials is treated with solvent so that the oil-bearing cells can release their contents. Commercial vegetable oils are refined in order to remove the non-glyceride impurities that are present in the crude oil or reduce them to a level where their deleterious effects on oil stability are minimum and made suitable for human consumption (Pal et al.,2015). Chemical refining includes degumming, neutralizing, bleaching, winterizing and deodorizing stages (Tasan and Demicri, 2005 ). Chemical refining decrease the oil yield, but has less effect on oil desirable components and improves oil stability (Suliman et al.,2013). Cold press oils considered as high-quality oils are defined as oils obtained only by mechanical means which are suitable for direct consumption and without heat treatment. In other words, cold pressed oil is generally ready for consumption without any need to be refined (Uitterhaegen and Evo, 2017). In the recent years, cold pressed GNO and crude GNO are also finding a lot of consumer interest at par with the refined GNO. Oil content and fatty acid composition of peanut have been studied in different cultivars and different environments (Hassan et al.,2005; Asibuo et al.,2008), but there is limited literature about the method of oil extraction process on fatty acid composition in GNO. While numerous studies have examined the fatty acid profile of GNO, comparative analyses of oils obtained through distinct extraction methods, particularly focusing on commercially refined, hexane-extracted crude, and cold-pressed oils, remain limited. This study aims to address this gap by investigating the differences in fatty acid composition among commercially refined groundnut oil (CRGNO), hexane-extracted crude groundnut oil (HCGNO), and cold-pressed groundnut oil (CPGNO). By elucidating the effects of these processing methods on the fatty acid profile, this research will yield pivotal findings, illuminating the optimal extraction methodologies to optimize the nutritional efficacy and organoleptic quality of groundnut oil, thereby contributing significantly to advancement of food science and technology. MATERIALS AND METHODS Procurement of raw materials Groundnut variety Kadiri 6 (50 Kgs) for the study (Fig. 1 .) was procured from Regional Agricultural Research Station, Palem, Professor Jayashankar Telangana State Agricultural University, Nagarkurnool District, Telangana and Agricultural Research Station, Kadiri, Acharya N G Ranga Agricultural University, Ananthapur District, Andhra Pradesh. Sample preparation The seed was cleaned and mixed homogeneously and pre-dried overnight at 40°C. Oil was extracted by two procedures viz., cold pressing and hexane extraction. For cold pressed oil, mechanical oil expression method was employed. A small capacity (10–20 kgh) table oil expeller (SP Engineering Corporation, Kanpur, India) was used for this purpose. The oil obtained had a considerable amount of foots (solids in suspension in expressed oil). These were removed through settling for 24 h followed by siphoning the top layer of oil leaving behind the sediments. The extracted cold pressed oil was stored at room temperature in glass containers, until their use in experiments. Solvent extraction method was carried out with hexane as a solvent for oil extraction from pulverized groundnut seeds. Commercial refined oil was sourced from Telangana State Oil Packaging Station in Shivarampally, Hyderabad. All the three oil samples were stored in glass bottles at room temperature prior to use for fatty acid profiling. Preparation of fatty acid methyl esters Fatty acid methyl esters from the three oil samples (cold pressed GNO, Hexane extracted crude GNO and Commercial refined GNO) was prepared by following procedure given by Geetha et al.,2016). 18.00g of KOH was weighed and added to 10ml of water, then diluted with methanol to 500ml, and allowed to stand for 24hours, and filtered into a polyethylene bottle. 50mg of oil was reconstituted in 1ml of 0.5M methanolic KOH and hydrolyzed at 80°C for 1 hour in a water bath. 1ml of fresh 10% BF3 in methanol was added and trans esterification was performed at 100°C for 20minutes in a water bath. After trans esterification, 2ml distilled water and 1ml hexane were added to the sample to quench the reaction. The organic phase was recovered by pipetting out the upper layer, pooled and spiked with the methyl ester. Fatty acid estimation using Gas Chromatography : Fatty acids were estimated as per methods given by Geetha et al . (2019) using GC, 7890B of Agilent Technologies with 7693 Auto sampler, equipped with flame ionization detector and split injector. Injector temperature was set at 260°C and samples were injected (1 µl) with split ratio of 10:1 by the auto sampler. Carrier gas (Nitrogen) flow rate was 30 ml/min. Column used was Agilent - DB-FFAP which is a nitroterephthalic-acid-modified polyethylene glycol (PEG) column of high polarity for the analysis of volatile fatty acids and phenols, with the length 30m X 250 µm, diameter 0.25mm, film thickness of 0.25µm was used. The temperature program was at set with the initial temperature of 100°C, hold time, 5 min, rising at an increasing rate to 240°C at the rate of 4°C /min and held for 5 min. Total run time was 45 min. Nitrogen was used as carrier gas at a column flow rate of 1.0 ml/min. Detector temperature was at 280°C. EZ Total Chrome software was used for running the GC and calculation of fatty acid composition. FID Hydrogen gas flow rate was 30 ml/min. Zero air flow was 300 ml/min and make up flow was 25 ml/min. The fatty acid content was measured based on area normalization. Standards used Standards used were 4 7885-U Supelco® 37 Component FAME Mix, 10 mg/mL in methylene chloride (varied), analytical standard. For individual trans-fatty acids standards, Supelco trans-9-Eliadic methyl ester, 10 mg/ml in heptane, trans-9, 12-Octadecadienoic (linoleliadic) methyl ester and trans-11-Vaccenic methyl ester, were used. After injecting the 37-FAME standard, individual trans-fatty acid methyl ester standards were also injected and the retention times were compared under standard conditions described above to ascertain that the individual standard peaks were coinciding exactly with the peaks in the combined standard. Samples were processed as described and injected as for standards. Sample fatty acid composition was compared with standard fatty acid composition and percentages calculated by normalization of peak areas. Clinical Trial Number : Not Applicable Statistical analysis All analyses were conducted in triplicate, and their mean and standard deviation were calculated. Significance between fatty acids was calculated and accepted at 95% confidence interval (p < 0.05) using one way ANOVA. Values were then expressed as Mean ± SD. RESULTS AND DISCUSSION i. Fatty acid composition chromatograms Groundnut oil is very common and one of the highly utilized vegetable oils in India, in both raw and refined forms for culinary applications like cooking and frying in Indian households (Asibuo et al.,2011). Dietary fats supply essential fatty acids which are required for maintenance of healthy skin, regulation of cholesterol metabolism, absorption of fat-soluble vitamins A, D, E, K and carotenoids (Bojang & Manchana, 2023 ). Fats provide the fatty acids which form structural components of biological membranes. Vegetable oils are important because of their high content in mono and polyunsaturated fatty acids when compared to animal fats (Amatsubo et al.,2006). Gas chromatography (GC) has been developed to determine the fatty acid composition and properties of fatty acids (FAs).The composition of FAs can be qualitatively and quantitatively analyzed on the basis of parameters such as retention time and peak area. Every type of edible oil has its own characteristic FAs, although some FAs are similar in various oils. The chain length and unsaturation degree are generally responsible for the principal variation in FAs of oils (Tian et al.,2019;Cui et al.,2017). Choice of solvents used for extraction of oil and also method of oil extraction plays a crucial role in the product outcome and FAs composition to some extent (Adebisi and Eriola, 2019 ). Abdolshahi et al., ( 2015 ) demonstrated that the choice of solvent of extraction influences fatty acid profile of pistachio oil. The fatty acid profile analysis of the three oils viz., CRGNO, HCGNO and CPGNO, is presented as the percentage peak area in the GC chromatograms (Figs. 2 , 3 & 4 ) and the fatty acid percentage is given in Table 1 . The total saturated fatty acid content was 15.48 ± 6.00%, 14.58 ± 5.68% and 15.93 ± 5.9% respectively in CPGNO, CRGNO and HCGNO. The total unsaturated fatty acid (UFAs) content in CPGNO, CRGNO and HCGNO was 84.74 ± 0.17%, 86.23 ± 0.23% and 85.12 ± 0.26% respectively, indicating high nutritional value, due to high UFAs (about 84–86%) that are essential to the human diet (Satil et al.,2003), the authors also highlighted that essential fatty acids like linolenic, linoleic and oleic acids have cardioprotective effects and ability to reduce the level of cholesterol in the blood. The U/S ratio in CPGNO, CRGNO and HCGNO was 5.47 ± 3.09, 5.91 ± 2.96 and 5.34 ± 3.11 respectively. The results of total saturated, unsaturated and U/S ratio did not have any significant differences. Similar observations that refining did not have significant effect on fatty acid compositions as found in the percentage peak area in the GC-MS chromatogram were reported by other researchers (Pal et al.,2015; Wronia et al.,2008). Table 1 Fatty acid composition of commercially available refined groundnut oil (CRGNO), hexane extracted crude groundnut oil (HCGNO) and cold pressed groundnut oil (CPGNO). Values are expressed as Mean ± SD. SFA - Saturated fatty acids, USFA - Unsaturated fatty acids, U/S Ratio - Unsaturated/Saturated fatty acid ratio, SE - Standard Error of mean, CD - Critical Difference Sample Saturated Fatty acids Unsaturated Fatty Acids Palmitic Acid (C16:0) Palmitoleic acid (C16:1) Stearic Acid (C18:0) Oleic Acid (C18:1n9cis) Linoleic Acid C18: 2n6cis Arachidic Acid (C20:0) Cis-11-Eicosenoic Acid (C20:1n9) Behenic Acid (C22:0) Lignoceric Acid (C24:0) SFA USFA U/S RATIO SE CD CPGNO 11.78 ± 0.04 0.09 ± 0.01 3.62 ± 0.06 40.22 ± 0.05 38.33 ± 0.03 1.46 ± 0.01 1.09 ± 0.02 2.54 ± 0.05 1 .00 ± 0.01 15.48 ± 6.00 84.74 ± 0.17 5.47 ± 3.09 0.06 0.2 CRGNO 11.16 ± 0.06 0.10 ± 0.01 3.32 ± 0.02 54.38 ± 0.06 26.41 ± 0.03 1.38 ± 0.04 1.11 ± 0.01 2.05 ± 0.04 0.90 ± 0.05 14.58 ± 5.68 86.23 ± 0.23 5.91 ± 2.96 0.14 0.47 HCGNO 11.78 ± 0.09 0.08 ± 0.01 4.07 ± 0.08 41.70 ± 0.05 36.90 ± 0.05 1.67 ± 0.04 1.14 ± 0.03 2.90 ± 0.01 0.81 ± 0.08 15.93 ± 5.95 85.12 ± 0.26 5.34 ± 3.11 0.16 0.53 It was observed that saturated fats like palmitic acid (C16:0), palmitoleic acid (C16:1) and stearic acid (C18:0) were almost similar in all the three oils. Other saturated fatty acids like caprylic acid (C8:0), capric acid (C10:0), lauric acid (C12:0), myristic acid (C14:0) and pentadecanoic acid (C15:0) were not detected in all the three oils. Among the unsaturated fats, CRGNO had higher oleic acid (C18:1n9cis) (54.38 ± 0.06%) and lower linoleic acid (C18:2n6cis) content (26.41 ± 0.03%), in comparison to CPGNO (40.22 ± 0.05% and 38.33 ± 0.03%) and HCGNO (41.70 ± 0.08% and 36.90 ± 0.05%), which could have been due to a possible mixing of oils in CRGNO. Oleic acid is a mono-unsaturated fatty acid (MUFA) essential to human nutrition and helps to reduce triglycerides, low-density cholesterol (LDL), total cholesterol and the glycemic index. Moreover, oleic acid is responsible for the increased stability and subdued oxidation of vegetable oil (Kocyigit et al., 2006 ). This is an indication that higher oleic acid content in CRGNO, could lead to better keeping quality of the oil compared to CPGNO and HCGNO. Despite the variations in oleic acid content, all three GNOs are a good source of oleic acid. Linoleic acid, an essential fatty acid of the omega-3 group, is very important for the development and maintenance of the nervous system;maintenance of growth, reproductive and physiological functions in humans since it reduces total and LDL-cholesterol levels (Sari et al.,2010). Linoleic acid (C18:2n6cis) was present in good amounts in all the three GNOs (38.33 ± 0.03% in CPGNO, 26.41 ± 0.03% in CRGNO and 36.90 ± 0.05% in HCGNO). Though the method of oil production did not change its composition of particular fatty acids, lower content of linoleic acid (C18:2n6cis) in CRGNO could possibly be due to the refining process, especially deodorization process. Cold pressed oils, virgin type oils and hot pressed oils, crude and bleached ones did not contain harmful trans isomers of fatty acids, however, there were 1.1% of them in the deodorised oils. Partly geometric isomerization of polyunsaturated acids (0.1% linoleic acid and1% linolenic acid) is caused by the process of deodorization and especially high temperature of up to 240°C during that process (Wagner et al.,2000). The stricter are the deodorization parameters (higher temperature, longer time) the higher is the content of trans isomers (Płatek and Krygier,1998), leading to reduction in the linoleic acid (C18:2n6cis). Apart from mixing of other oils, deodorization step in refining process could slightly lower the linoleic acid (C18:2n6cis) content in refined oils. Similar results were obtained in our study with lower linoleic acid (C18:2n6cis) in CRGNO, compared to CPGNO and HCGNO. Method of oil extraction had an influence on linoleic acid (C18:2n6cis) content, and also on the total essential fatty acid content, indicating that method of oil extraction does have a mild impact on the essential fatty acid content in the oils. Other unsaturated fatty acids like arachidic acid (C20:0) and behenic acid (C22:0) were slightly lower in CRGNO when compared with CPGNO and HCGNO. Cis-11-Eicosenoic acid (C20:1n9) was slightly higher in the HCGNO when compared with CRGNO and CPGNO. However, the differences were not statistically significant. Lignoceric acid (C24:0) was present in traces in all the three GNOs (0.81 ± 0.08% to 1.00 ± 0.01%). Lignoceric acid, a byproduct of lignin production is present in peanut oil in small amounts (1.1% − 2.2%), and functions as an enzyme in the nervous system to produce 2-hydroxy fatty acids that are major components of myelin lipids (Beare et al.,2009). Although the fatty acid composition of groundnut oil is primarily governed by the genetic traits of the seed, our findings suggest that the extraction and refining processes can affect the relative levels of certain fatty acids. Specifically, cold-pressed and hexane-extracted oils retained higher linoleic acid (C18:2n6cis) content compared to refined oil, likely due to minimal thermal or chemical processing. Several authors have investigated the influence of industrial processing, especially the refining process on quality and stability of different vegetable oils (Pal et al.,2015;Verhe et al.,2006). Moreover, in the present study, our primary objective was to assess the effect of oil extraction methods—cold pressing, hexane extraction, and commercial refining on the fatty acid composition. To control for varietal differences, both the cold-pressed groundnut oil (CPGNO) and the hexane-extracted groundnut oil (HCGNO) were produced from the same batch of groundnuts, thereby ensuring a uniform genetic source for these two treatments. Effect of pressing conditions on non-refined sunflower oil has been presented by Turkulov et al. ( 1998 ) and the influence of water degumming in phosphatide content has been discussed by Crapiste et al. ( 1998 ). Vegetable oils are used for many food and industrial purposes, and the oil quality is associated with its FA composition. Therefore fatty acid composition of edible oils is also dependent on the plant variety, degree of ripening seeds, and climatic conditions. Several studies have documented that temperature is the predominant environmental factor affecting FA composition, especially for unsaturated fatty acids (UFAs) (Varnham, 2015 ; Jedidi et al.,2020).The results of our study indicate that method of oil extraction also has an influence on the fatty acid composition of the oils. Mezzomo et al. ( 2010 ) reported that the oil extraction techniques used to obtain high aggregate value compounds from natural products are critical for product quality. The three GNOs showed few variations with either increase or decrease in fatty acid composition from between the hexane extracted crude form to refined form to cold pressed oil. However, the significant variation was observed only with oleic acid (C18:1n9cis) and linoleic acid (C18:2n6cis) content. Abdolshahi et al. ( 2015 ) outlined that saturated and unsaturated fatty acid profiles of pistachio oil extracted by Soxhlet and maceration method were not different but their content was statistically different. According to Achinewhu and Akpapunam, ( 2005 ) refining does not have much effect on fatty acid composition except for some slight inconsistent decrease in saturated and unsaturated fatty acid. Aluyor et al. ( 2009 ) also reported that refining did not have much effect on fatty acid composition and that there was no undesirable polymerization of oil during the refining process. In contrast to above findings, our study indicates that method of oil extraction does have a mild impact on the essential fatty acid content in the oils. Cold pressed GNO and hexane extracted crude GNO had higher essential fatty acid contentlike linoleic acid (C18:2n6cis), as compared to refined GNO. One of the key strengths of this study lies in its controlled experimental design, particularly the use of a single groundnut batch for both cold-pressed and hexane-extracted oils, which allowed for a focused evaluation of how extraction methods influence fatty acid composition while minimizing genotypic variation. Additionally, the integration of a commercially refined oil sample offers practical relevance, reflecting products typically available to consumers. However, the commercially refined groundnut oil (CRGNO) used in this study was obtained as a representative sample from the retail market, and its varietal origin was not disclosed by the manufacturer. As such, while the CRGNO sample offers practical relevance for comparison from a consumer perspective, we acknowledge that its unknown genotype may introduce variability unrelated to the extraction process. This limitation highlights the importance of sourcing oils from known and controlled groundnut varieties in future studies, or alternatively, comparing a range of commercial oils to better account for this variability. Despite this, the findings reveal subtle but meaningful variations in essential fatty acid content especially linoleic acid,associated with different extraction and refining techniques, suggesting that processing conditions can influence nutritional quality to a modest extent. As such, this study contributes new insight into the nuanced impact of oil processing on fatty acid profiles, an area that has received limited attention in groundnut oil research. Future investigations would benefit from including multiple groundnut varieties with clearly documented genetic backgrounds, as well as a broader spectrum of commercial oils, to more comprehensively assess the interplay between genotype and processing. Collectively, this work augments our understanding of the relationship between extraction method and oil quality, offering beneficial implications for both industry practices, consumer safety and public health nutrition. CONCLUSION The comparative analysis of fatty acid profiles across commercially refined, cold-pressed, and hexane-extracted groundnut oils revealed no statistically significant differences in the overall proportions of total saturated fatty acids, total unsaturated fatty acids, or the unsaturated-to-saturated (U/S) ratio. However, a significant modulation of individual fatty acid components was observed. Specifically, commercially refined groundnut oil exhibited a marked increase in oleic acid (C18:1n9cis) and a decrease in linoleic acid (C18:2n6cis). These results demonstrate that the oil extraction methodology exerts a discernible influence on the retention of essential fatty acids. Notably, cold-pressed and hexane-extracted crude groundnut oils presented a higher linoleic acid (C18:2n6cis) content compared to the refined counterpart. Conversely, the increased oleic acid (C18:1n9cis) levels in the commercially refined oil suggest an enhanced oxidative stability, potentially translating to improved shelf life relative to the cold-pressed and hexane-extracted oils. This study accentuate the critical impact of extraction processes on the final fatty acid composition of groundnut oil, with direct implications for its nutritional value and oxidative stability. These findings highlights the subtle yet important ways in which extraction methods shape the nutritional and functional properties of edible oils. The study contributes to the growing body of evidence that, beyond the seed genotype, post-harvest processing steps exert a meaningful influence on oil quality. From a nutritional science perspective, this highlights the trade-offs between stability and essential fatty acid retention when choosing oil processing methods. The insights gained here have practical relevance for both food manufacturers aiming to optimize oil quality for specific culinary uses and for health-conscious consumers seeking oils with superior nutritional profiles. Moreover, this research emphasizes the need for greater transparency in labeling and sourcing, particularly as demand for minimally processed and functional foods continues to rise. Declarations Author Contribution KBP: Conceptualization, Data curation, Resource Acquisition, Statistical analysis, Investigation, Methodology, Writing original draft. AK.: Conceptualization, Data curation, Methodology,, Supervision, Visualization, Writing original draft. Supervision and validation of results. All authors reviewed the manuscript. Acknowledgement The authors gratefully acknowledge the Regional Agricultural Research Station, Palem, Professor Jayashankar Telangana State Agricultural University, Nagarkurnool District, Telangana, and the Agricultural Research Station, Kadiri, Acharya N G Ranga Agricultural University, Ananthapur District, Andhra Pradesh, for providing the Kadiri 6 groundnut variety used in this study. We extend our sincere thanks to the MFPI-Quality Control Laboratory, PJTS Agricultural University, Hyderabad, for their support and assistance in the analysis. 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Afr J Biotechnol, 7(13): 2203–2208 Geetha S, Manorama K, Aparna K, Jagan CH (2016) Determination of fatty acid profile of branded and unbranded processed foods commonly available in the Indian market with special reference to trans fatty acids. J Oilseeds Res 33(3):168–173 Ogunsina BS, Olaoye IO, Adegbenjo AO, Babawale BD (2011) Nutritional and physical properties of kariya seeds. Int Agrophys 25:97–100 Bojang KP, Manchana V (2023) Nutrition and Healthy Aging: A Review. Curr Nutr Rep 12(3):369–375. https://doi.org/10.1007/s13668-023-00473-0 Amatsubo T, Hagura Y, Suzuki K (2006) The effect of superheated steam treatment on the quality of vegetable oils. J Food Sci Technol Res 12(2):114–118 Tian L, Zeng Y, Zheng X, Chiu Y, Liu T (2019) Detection of Peanut Oil Adulteration Mixed with Rapeseed Oil Using Gas Chromatography and Gas Chromatography–Ion Mobility Spectrometry. Food Anal Methods 12:2282–2292 Cui Y, Hao P, Liu B, Meng X (2017) Effect of traditional Chinese cooking methods on fatty acid profiles of vegetable oils. Food Chem 233:77–84 Abdolshahi A, Majd M, Rad JS, Taheri M, Shabani A, Da Silva JAT (2015) Choice of solvent extraction technique affects fatty acid composition of pistachio ( Pistacia vera L.) oil. J Food Sci Technol 52:422–2427 Satil F, Ayas N, Baser K, Husnu C (2003) Fatty Acid Composition of Pistachio Nuts in Turkey. Chem Nat Compd 39:322–324 Wronia M, Krygier K, Kaczmarczyk M (2008) Comparison of the quality of cold pressed and virgin rapeseed oils with industrially obtained oils. Pol J Food Nutr Sci 58:1 Kocyigit A, Koylu AA, Keles H (2006) Effects of pistachio nuts consumption on plasma lipid profile and oxidative status in healthy volunteers. Nutr Metabolism Cardiovasc Disease 16(3):202–209 Sari I, Baltaci Y, Bagci C, Davutoglu V, Erel O, Celik H, Ozer O, Aksoy N, Aksoy M (2010) Effect of pistachio diet on lipid parameters, endothelial function, inflammation, and oxidative status: a prospective study. Nutrition 26:399–404 Wagner K, Auer E, Elmadfa I (2000) Content of trans fatty acids in margarines, plant oils, fried products and chocolate spreads in Austria. Eur Food Res Technol 210:237–241 Płatek T, Krygier K (1998) Characteristics of rapeseed oils after refining processes in industrial scale. Roczniki IPMiT 35:171–173 Beare RJL, Dieffenbacher A, Holm JV (2009) Lexicon of lipid nutrition (IUPAC Technical Report). Pure and Applied Chemistry , 73(4): 685–744 Verhe R, Verleyen T, Van Hoed V, De Greyt W (2006) Influence of refining of vegetable oils on minor components. J Oil Palm Res 1:168–179 Turkulov J, Dimic E, Karlovic D, Vuksa V (1998) Effect of hydrothermal treatment on the quality of nonrefined edible sunflower oil- In World Conference on Oilseed and Edible Oil Processing . Vol I, pp 185–187, S.S. Koseoglu, K.C. Rhee and R. F. Wilson (Ed.). AOCS Press, Champaign, IL Crapiste GH, Brevedan M, Carelli AA (1998) Water degumming of sunflower oils- In World Conference on Oilseed and Edible Oil Processing . Vol II, pp 32–35, S.S. Koseoglu, K.C. Rhee and R. F. Wilson (Ed.). AOCS Press, Champaign, IL Varnham A (2015) Seed Oil: Biological Properties, Health Benefits and Commercial Applications. Nova Science Publishers Inc, New York Jedidi B, Mokbli S, Sbihi SM, Nehdi IA, Romdhani-Younes M, Al-Resayes SI (2020) Effect of extraction solvents on fatty acid composition and physicochemical properties of Tecoma stans seed oils. J King Saud Univ – Sci 32:2468–2473 Mezzomo N, Mileo B, Friedrich M, Martínez J, Ferreira S (2010) Supercritical fluid extraction of peach ( Prunus persica ) almond oil: Process yield and extract composition. Bioresour Technol 101(14):5622–5632 Achinewhu SC, Akpapunam MA (2005) Physical and chemical characteristics of refined vegetable oils from rubber seed and bread fruit. J Plant Food 35:103–107 Aluyor EO, Aluyor P, Ozigagu CE (2009) Effect of refining on the quality and composition of groundnut oil. Afr J Food Sci 3(8):201–205 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Revision requested 20 Dec, 2025 Reviews received at journal 19 Dec, 2025 Reviewers agreed at journal 17 Dec, 2025 Reviews received at journal 24 Oct, 2025 Reviewers agreed at journal 15 Oct, 2025 Reviewers agreed at journal 12 Oct, 2025 Reviewers invited by journal 20 Aug, 2025 Editor assigned by journal 17 Jul, 2025 Submission checks completed at journal 17 Jul, 2025 First submitted to journal 16 Jul, 2025 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. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7142039","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":503435480,"identity":"3987c4f7-17ff-4566-aa7f-21d3d7f99fd3","order_by":0,"name":"Keotshepile Precious Bojang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABAElEQVRIiWNgGAWjYLCCBCA2gFA2QMzYeIAULWkgLQ2EtTBAtIDAYTCJV4u59OGDHx5UHLY3Z294+Lng13m7te2HgbbU2ETj0mLZl5YskXDmcOLOngPJ0jP7bidvO5MI1HIsLbcBl3vO8JgxJLYdTjC4kZAgzdtzO9nsAFALY8NhPFr4v4G02AO1JP/m7TmXbHb+ISEtPGwgLYwbbiSkSfP8OGBndoOALZY9bMZAv6SD/JJmzduQnGB2A2hLAh6/mPMwP/z4o8IaGGI9ybd5/tjZm51Pf/jgQ40NbochmDwJDIxtDIlglQk4lKNpYT/AwPCHwR6P4lEwCkbBKBihAAB3+Gd23TvrlgAAAABJRU5ErkJggg==","orcid":"","institution":"Professor Jayashankar Telangana State Agricultural University","correspondingAuthor":true,"prefix":"","firstName":"Keotshepile","middleName":"Precious","lastName":"Bojang","suffix":""},{"id":503435481,"identity":"8c19d006-466f-4714-8c5e-3bab1e80021e","order_by":1,"name":"Aparna Kuna","email":"","orcid":"","institution":"Professor Jayashankar Telangana State Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Aparna","middleName":"","lastName":"Kuna","suffix":""}],"badges":[],"createdAt":"2025-07-16 16:23:24","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7142039/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7142039/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":90204422,"identity":"4944b464-3480-49ff-81d8-91c717f9076a","added_by":"auto","created_at":"2025-08-29 20:32:04","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":209682,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGroundnut seed Kadiri 6 variety\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7142039/v1/5d951a067cfa526a5642c245.png"},{"id":90204427,"identity":"ccb02e6a-074f-42b1-a531-1afd6f8d4028","added_by":"auto","created_at":"2025-08-29 20:32:04","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":48297,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFatty Acid Composition in cold pressed groundnut oil\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7142039/v1/793f06636aab26b79ab2e935.jpg"},{"id":90204764,"identity":"f6fd3546-6ce0-4a4c-87a3-5b05d1df491f","added_by":"auto","created_at":"2025-08-29 20:40:04","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":48682,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFatty Acid composition of commercial groundnut oil\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7142039/v1/5eab9e3b8637b382652b76cb.jpg"},{"id":90204424,"identity":"cee36732-24a2-411d-964f-5293475f603b","added_by":"auto","created_at":"2025-08-29 20:32:04","extension":"jpeg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":215681,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFatty acid composition of hexane extracted groundnut oil\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7142039/v1/b29a52855dfabbd04d4fccbb.jpeg"},{"id":90205260,"identity":"d0264fc0-0c0d-4ba0-bfe7-417b8b77d0e5","added_by":"auto","created_at":"2025-08-29 20:56:04","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1204826,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7142039/v1/5fe033ae-76e6-43ab-baf3-8136c1576c09.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eEffect of Oil Extraction Process on the Fatty Acid Composition of Groundnut Oil\u003c/p\u003e","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eThe legume \u003cem\u003eArachis hypogaea\u003c/em\u003e, commonly known as peanut or groundnut, is a very important food crop throughout the tropics and subtropics. Groundnut is one of the most widely used legumes for its nutrition and taste, and occupies a rank of major oilseed crop in the world, due to its role in health promoting effect. Groundnut oil (GNO) owing to its unique organoleptic properties associated with its cardioprotective and anti-inflammatory properties has found its place on the highly competitive international edible oil market (Shamim et al.,2014). Its versatile applications span culinary realms, ranging from direct consumption to the production of cooking oil and confectionery products. Beyond its gastronomic significance, groundnut is renowned for its rich composition of bioactive compounds, including tocopherols (vitamin E) and phytosterols, notably β-sitosterol, which confer antioxidant and cholesterol-lowering properties (Maestri, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2024\u003c/span\u003e;Shin et al.,2009). The primary economic value of groundnut lies in its oil content, which typically ranges from 40\u0026ndash;50% of the kernel weight, making it a substantial source of vegetable oil worldwide (Janila, 2016).\u003c/p\u003e\u003cp\u003eEdible oils are often subject to intentional or unintentional mixing, such as blending cold-pressed oils with solvent-extracted or refined variants, which is frequently done to reduce production costs or enhance yield. This practice raises concerns over authenticity and quality, particularly when higher value oils are replaced with cheaper alternatives (Bojang et al.,2021). For example, GNO is blended with ricebran oil, sunflower oil, soybean oil, palm oil, or canola oil in varying proportions, or adulterated by adding its flavor to cheaper oils to fake it (Jee,2009 ;Fang et al.,2013;Xie et al.,2013).The main compound in vegetable oil is triglyceride (\u0026gt;\u0026thinsp;95%), which is composed of fatty acids (Ai et al.,2009). Fatty acid profiles can serve as a unique characteristic feature of edible vegetable oil. Fatty acids composition of vegetable oils is formed by a mixture of saturated and unsaturated fatty acids classified according to the number of unsaturated bonds as monounsaturated (MUFAs) or polyunsaturated fatty acids (PUFAs).\u003c/p\u003e\u003cp\u003eThe fatty acid profile of groundnut oil (GNO) is a critical determinant of its nutritional quality and oxidative stability. Predominantly, GNO is characterized by a high proportion of unsaturated fatty acids, particularly oleic acid (C18:1n9cis) and linoleic acid (C18:2n6cis), which collectively constitute over 80% of the total fatty acids (Musalima et al.,2019).These unsaturated fatty acids are essential for human health, playing vital roles in cell membrane integrity, eicosanoid synthesis, and cholesterol metabolism (Simopoulos et al.,2016).\u003c/p\u003e\u003cp\u003eTo extract oil from seeds, fruits and other oil-bearing materials, different extraction methods are used, for example cold pressing, use of polar and non-polar solvents depending on the intended use (Nagaraj, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2009\u003c/span\u003e;Adebisi and Eriola,2019;Adebayo et al.,2015). The extraction method employed to obtain GNO significantly influences its final composition and quality. However, most of the time commercial mills employs a combination of various unit operations including conditioning, flaking and solvent extraction whereby the oil-bearing materials is treated with solvent so that the oil-bearing cells can release their contents. Commercial vegetable oils are refined in order to remove the non-glyceride impurities that are present in the crude oil or reduce them to a level where their deleterious effects on oil stability are minimum and made suitable for human consumption (Pal et al.,2015). Chemical refining includes degumming, neutralizing, bleaching, winterizing and deodorizing stages (Tasan and Demicri, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). Chemical refining decrease the oil yield, but has less effect on oil desirable components and improves oil stability (Suliman et al.,2013). Cold press oils considered as high-quality oils are defined as oils obtained only by mechanical means which are suitable for direct consumption and without heat treatment. In other words, cold pressed oil is generally ready for consumption without any need to be refined (Uitterhaegen and Evo, 2017).\u003c/p\u003e\u003cp\u003eIn the recent years, cold pressed GNO and crude GNO are also finding a lot of consumer interest at par with the refined GNO. Oil content and fatty acid composition of peanut have been studied in different cultivars and different environments (Hassan et al.,2005; Asibuo et al.,2008), but there is limited literature about the method of oil extraction process on fatty acid composition in GNO. While numerous studies have examined the fatty acid profile of GNO, comparative analyses of oils obtained through distinct extraction methods, particularly focusing on commercially refined, hexane-extracted crude, and cold-pressed oils, remain limited. This study aims to address this gap by investigating the differences in fatty acid composition among commercially refined groundnut oil (CRGNO), hexane-extracted crude groundnut oil (HCGNO), and cold-pressed groundnut oil (CPGNO). By elucidating the effects of these processing methods on the fatty acid profile, this research will yield pivotal findings, illuminating the optimal extraction methodologies to optimize the nutritional efficacy and organoleptic quality of groundnut oil, thereby contributing significantly to advancement of food science and technology.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003e\u003cstrong\u003eProcurement of raw materials\u003c/strong\u003e\u003cp\u003eGroundnut variety Kadiri 6 (50 Kgs) for the study (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.) was procured from Regional Agricultural Research Station, Palem, Professor Jayashankar Telangana State Agricultural University, Nagarkurnool District, Telangana and Agricultural Research Station, Kadiri, Acharya N G Ranga Agricultural University, Ananthapur District, Andhra Pradesh.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eSample preparation\u003c/strong\u003e\u003cp\u003eThe seed was cleaned and mixed homogeneously and pre-dried overnight at 40\u0026deg;C. Oil was extracted by two procedures viz., cold pressing and hexane extraction. For cold pressed oil, mechanical oil expression method was employed. A small capacity (10\u0026ndash;20 kgh) table oil expeller (SP Engineering Corporation, Kanpur, India) was used for this purpose. The oil obtained had a considerable amount of foots (solids in suspension in expressed oil). These were removed through settling for 24 h followed by siphoning the top layer of oil leaving behind the sediments. The extracted cold pressed oil was stored at room temperature in glass containers, until their use in experiments. Solvent extraction method was carried out with hexane as a solvent for oil extraction from pulverized groundnut seeds. Commercial refined oil was sourced from Telangana State Oil Packaging Station in Shivarampally, Hyderabad. All the three oil samples were stored in glass bottles at room temperature prior to use for fatty acid profiling.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003ePreparation of fatty acid methyl esters\u003c/strong\u003e\u003cp\u003eFatty acid methyl esters from the three oil samples (cold pressed GNO, Hexane extracted crude GNO and Commercial refined GNO) was prepared by following procedure given by Geetha et al.,2016). 18.00g of KOH was weighed and added to 10ml of water, then diluted with methanol to 500ml, and allowed to stand for 24hours, and filtered into a polyethylene bottle. 50mg of oil was reconstituted in 1ml of 0.5M methanolic KOH and hydrolyzed at 80\u0026deg;C for 1 hour in a water bath. 1ml of fresh 10% BF3 in methanol was added and \u003cem\u003etrans\u003c/em\u003e esterification was performed at 100\u0026deg;C for 20minutes in a water bath. After trans esterification, 2ml distilled water and 1ml hexane were added to the sample to quench the reaction. The organic phase was recovered by pipetting out the upper layer, pooled and spiked with the methyl ester.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eFatty acid estimation using Gas Chromatography\u003c/b\u003e: Fatty acids were estimated as per methods given by Geetha \u003cem\u003eet al\u003c/em\u003e. (2019) using GC, 7890B of Agilent Technologies with 7693 Auto sampler, equipped with flame ionization detector and split injector. Injector temperature was set at 260\u0026deg;C and samples were injected (1 \u0026micro;l) with split ratio of 10:1 by the auto sampler. Carrier gas (Nitrogen) flow rate was 30 ml/min. Column used was Agilent - DB-FFAP which is a nitroterephthalic-acid-modified polyethylene glycol (PEG) column of high polarity for the analysis of volatile fatty acids and phenols, with the length 30m X 250 \u0026micro;m, diameter 0.25mm, film thickness of 0.25\u0026micro;m was used. The temperature program was at set with the initial temperature of 100\u0026deg;C, hold time, 5 min, rising at an increasing rate to 240\u0026deg;C at the rate of 4\u0026deg;C /min and held for 5 min. Total run time was 45 min. Nitrogen was used as carrier gas at a column flow rate of 1.0 ml/min. Detector temperature was at 280\u0026deg;C. EZ Total Chrome software was used for running the GC and calculation of fatty acid composition. FID Hydrogen gas flow rate was 30 ml/min. Zero air flow was 300 ml/min and make up flow was 25 ml/min. The fatty acid content was measured based on area normalization.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eStandards used\u003c/strong\u003e\u003cp\u003eStandards used were 4\u003cem\u003e7885-U\u003c/em\u003e Supelco\u0026reg; 37 Component FAME Mix, 10 mg/mL in methylene chloride (varied), analytical standard. For individual trans-fatty acids standards, Supelco trans-9-Eliadic methyl ester, 10 mg/ml in heptane, trans-9, 12-Octadecadienoic (linoleliadic) methyl ester and trans-11-Vaccenic methyl ester, were used. After injecting the 37-FAME standard, individual trans-fatty acid methyl ester standards were also injected and the retention times were compared under standard conditions described above to ascertain that the individual standard peaks were coinciding exactly with the peaks in the combined standard. Samples were processed as described and injected as for standards. Sample fatty acid composition was compared with standard fatty acid composition and percentages calculated by normalization of peak areas.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eClinical Trial Number\u003c/b\u003e : Not Applicable\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eStatistical analysis\u003c/strong\u003e\u003cp\u003eAll analyses were conducted in triplicate, and their mean and standard deviation were calculated. Significance between fatty acids was calculated and accepted at 95% confidence interval (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) using one way ANOVA. Values were then expressed as Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD.\u003c/p\u003e\u003c/p\u003e"},{"header":"RESULTS AND DISCUSSION","content":"\u003cp\u003e\u003cb\u003ei. Fatty acid composition chromatograms\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eGroundnut oil is very common and one of the highly utilized vegetable oils in India, in both raw and refined forms for culinary applications like cooking and frying in Indian households (Asibuo et al.,2011). Dietary fats supply essential fatty acids which are required for maintenance of healthy skin, regulation of cholesterol metabolism, absorption of fat-soluble vitamins A, D, E, K and carotenoids (Bojang \u0026amp; Manchana, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Fats provide the fatty acids which form structural components of biological membranes. Vegetable oils are important because of their high content in mono and polyunsaturated fatty acids when compared to animal fats (Amatsubo et al.,2006).\u003c/p\u003e\u003cp\u003eGas chromatography (GC) has been developed to determine the fatty acid composition and properties of fatty acids (FAs).The composition of FAs can be qualitatively and quantitatively analyzed on the basis of parameters such as retention time and peak area. Every type of edible oil has its own characteristic FAs, although some FAs are similar in various oils. The chain length and unsaturation degree are generally responsible for the principal variation in FAs of oils (Tian et al.,2019;Cui et al.,2017). Choice of solvents used for extraction of oil and also method of oil extraction plays a crucial role in the product outcome and FAs composition to some extent (Adebisi and Eriola, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Abdolshahi et al., (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) demonstrated that the choice of solvent of extraction influences fatty acid profile of pistachio oil.\u003c/p\u003e\u003cp\u003eThe fatty acid profile analysis of the three oils viz., CRGNO, HCGNO and CPGNO, is presented as the percentage peak area in the GC chromatograms (Figs.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e \u0026amp; \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e) and the fatty acid percentage is given in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The total saturated fatty acid content was 15.48\u0026thinsp;\u0026plusmn;\u0026thinsp;6.00%, 14.58\u0026thinsp;\u0026plusmn;\u0026thinsp;5.68% and 15.93\u0026thinsp;\u0026plusmn;\u0026thinsp;5.9% respectively in CPGNO, CRGNO and HCGNO. The total unsaturated fatty acid (UFAs) content in CPGNO, CRGNO and HCGNO was 84.74\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17%, 86.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23% and 85.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26% respectively, indicating high nutritional value, due to high UFAs (about 84\u0026ndash;86%) that are essential to the human diet (Satil et al.,2003), the authors also highlighted that essential fatty acids like linolenic, linoleic and oleic acids have cardioprotective effects and ability to reduce the level of cholesterol in the blood. The U/S ratio in CPGNO, CRGNO and HCGNO was 5.47\u0026thinsp;\u0026plusmn;\u0026thinsp;3.09, 5.91\u0026thinsp;\u0026plusmn;\u0026thinsp;2.96 and 5.34\u0026thinsp;\u0026plusmn;\u0026thinsp;3.11 respectively. The results of total saturated, unsaturated and U/S ratio did not have any significant differences. Similar observations that refining did not have significant effect on fatty acid compositions as found in the percentage peak area in the GC-MS chromatogram were reported by other researchers (Pal et al.,2015; Wronia et al.,2008).\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\u003eFatty acid composition of commercially available refined groundnut oil (CRGNO), hexane extracted crude groundnut oil (HCGNO) and cold pressed groundnut oil (CPGNO). Values are expressed as Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD. SFA - Saturated fatty acids, USFA - Unsaturated fatty acids, U/S Ratio - Unsaturated/Saturated fatty acid ratio, SE - Standard Error of mean, CD - Critical Difference\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"15\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c14\" colnum=\"14\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c15\" colnum=\"15\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eSample\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e\u003cp\u003eSaturated Fatty acids\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"6\" nameend=\"c10\" namest=\"c5\"\u003e\u003cp\u003eUnsaturated Fatty Acids\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePalmitic Acid (C16:0)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePalmitoleic acid (C16:1)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eStearic Acid (C18:0)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eOleic Acid (C18:1n9cis)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eLinoleic\u003c/p\u003e\u003cp\u003eAcid\u003c/p\u003e\u003cp\u003eC18: 2n6cis\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eArachidic Acid (C20:0)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eCis-11-Eicosenoic Acid (C20:1n9)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eBehenic Acid (C22:0)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c10\"\u003e\u003cp\u003eLignoceric Acid (C24:0)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c11\"\u003e\u003cp\u003eSFA\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c12\"\u003e\u003cp\u003eUSFA\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c13\"\u003e\u003cp\u003eU/S RATIO\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c14\"\u003e\u003cp\u003eSE\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c15\"\u003e\u003cp\u003eCD\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCPGNO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e11.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e0.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e3.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e40.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e38.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e1.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c8\"\u003e\u003cp\u003e1.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c9\"\u003e\u003cp\u003e2.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c10\"\u003e\u003cp\u003e1 .00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c11\"\u003e\u003cp\u003e15.48\u0026thinsp;\u0026plusmn;\u0026thinsp;6.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c12\"\u003e\u003cp\u003e84.74 \u0026plusmn; 0.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c13\"\u003e\u003cp\u003e5.47 \u0026plusmn; 3.09\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e\u003cp\u003e0.06\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e\u003cp\u003e0.2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCRGNO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e11.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e0.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e3.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e54.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e26.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e1.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c8\"\u003e\u003cp\u003e1.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c9\"\u003e\u003cp\u003e2.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c10\"\u003e\u003cp\u003e0.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c11\"\u003e\u003cp\u003e14.58\u0026thinsp;\u0026plusmn;\u0026thinsp;5.68\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c12\"\u003e\u003cp\u003e86.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c13\"\u003e\u003cp\u003e5.91\u0026thinsp;\u0026plusmn;\u0026thinsp;2.96\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e\u003cp\u003e0.14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e\u003cp\u003e0.47\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHCGNO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e11.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e0.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e4.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e41.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e36.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e1.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c8\"\u003e\u003cp\u003e1.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c9\"\u003e\u003cp\u003e2.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c10\"\u003e\u003cp\u003e0.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c11\"\u003e\u003cp\u003e15.93\u0026thinsp;\u0026plusmn;\u0026thinsp;5.95\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c12\"\u003e\u003cp\u003e85.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c13\"\u003e\u003cp\u003e5.34\u0026thinsp;\u0026plusmn;\u0026thinsp;3.11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e\u003cp\u003e0.16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e\u003cp\u003e0.53\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\u003eIt was observed that saturated fats like palmitic acid (C16:0), palmitoleic acid (C16:1) and stearic acid (C18:0) were almost similar in all the three oils. Other saturated fatty acids like caprylic acid (C8:0), capric acid (C10:0), lauric acid (C12:0), myristic acid (C14:0) and pentadecanoic acid (C15:0) were not detected in all the three oils.\u003c/p\u003e\u003cp\u003eAmong the unsaturated fats, CRGNO had higher oleic acid (C18:1n9cis) (54.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06%) and lower linoleic acid (C18:2n6cis) content (26.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03%), in comparison to CPGNO (40.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05% and 38.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03%) and HCGNO (41.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08% and 36.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05%), which could have been due to a possible mixing of oils in CRGNO. Oleic acid is a mono-unsaturated fatty acid (MUFA) essential to human nutrition and helps to reduce triglycerides, low-density cholesterol (LDL), total cholesterol and the glycemic index. Moreover, oleic acid is responsible for the increased stability and subdued oxidation of vegetable oil (Kocyigit et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). This is an indication that higher oleic acid content in CRGNO, could lead to better keeping quality of the oil compared to CPGNO and HCGNO. Despite the variations in oleic acid content, all three GNOs are a good source of oleic acid. Linoleic acid, an essential fatty acid of the omega-3 group, is very important for the development and maintenance of the nervous system;maintenance of growth, reproductive and physiological functions in humans since it reduces total and LDL-cholesterol levels (Sari et al.,2010). Linoleic acid (C18:2n6cis) was present in good amounts in all the three GNOs (38.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03% in CPGNO, 26.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03% in CRGNO and 36.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05% in HCGNO).\u003c/p\u003e\u003cp\u003eThough the method of oil production did not change its composition of particular fatty acids, lower content of linoleic acid (C18:2n6cis) in CRGNO could possibly be due to the refining process, especially deodorization process. Cold pressed oils, virgin type oils and hot pressed oils, crude and bleached ones did not contain harmful trans isomers of fatty acids, however, there were 1.1% of them in the deodorised oils. Partly geometric isomerization of polyunsaturated acids (0.1% linoleic acid and1% linolenic acid) is caused by the process of deodorization and especially high temperature of up to 240\u0026deg;C during that process (Wagner et al.,2000). The stricter are the deodorization parameters (higher temperature, longer time) the higher is the content of trans isomers (Płatek and Krygier,1998), leading to reduction in the linoleic acid (C18:2n6cis). Apart from mixing of other oils, deodorization step in refining process could slightly lower the linoleic acid (C18:2n6cis) content in refined oils. Similar results were obtained in our study with lower linoleic acid (C18:2n6cis) in CRGNO, compared to CPGNO and HCGNO. Method of oil extraction had an influence on linoleic acid (C18:2n6cis) content, and also on the total essential fatty acid content, indicating that method of oil extraction does have a mild impact on the essential fatty acid content in the oils.\u003c/p\u003e\u003cp\u003eOther unsaturated fatty acids like arachidic acid (C20:0) and behenic acid (C22:0) were slightly lower in CRGNO when compared with CPGNO and HCGNO. Cis-11-Eicosenoic acid (C20:1n9) was slightly higher in the HCGNO when compared with CRGNO and CPGNO. However, the differences were not statistically significant. Lignoceric acid (C24:0) was present in traces in all the three GNOs (0.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08% to 1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01%). Lignoceric acid, a byproduct of lignin production is present in peanut oil in small amounts (1.1% \u0026minus;\u0026thinsp;2.2%), and functions as an enzyme in the nervous system to produce 2-hydroxy fatty acids that are major components of myelin lipids (Beare et al.,2009).\u003c/p\u003e\u003cp\u003eAlthough the fatty acid composition of groundnut oil is primarily governed by the genetic traits of the seed, our findings suggest that the extraction and refining processes can affect the relative levels of certain fatty acids. Specifically, cold-pressed and hexane-extracted oils retained higher linoleic acid (C18:2n6cis) content compared to refined oil, likely due to minimal thermal or chemical processing. Several authors have investigated the influence of industrial processing, especially the refining process on quality and stability of different vegetable oils (Pal et al.,2015;Verhe et al.,2006). Moreover, in the present study, our primary objective was to assess the effect of oil extraction methods\u0026mdash;cold pressing, hexane extraction, and commercial refining on the fatty acid composition. To control for varietal differences, both the cold-pressed groundnut oil (CPGNO) and the hexane-extracted groundnut oil (HCGNO) were produced from the same batch of groundnuts, thereby ensuring a uniform genetic source for these two treatments.\u003c/p\u003e\u003cp\u003eEffect of pressing conditions on non-refined sunflower oil has been presented by Turkulov et al. (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e1998\u003c/span\u003e) and the influence of water degumming in phosphatide content has been discussed by Crapiste et al. (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e1998\u003c/span\u003e). Vegetable oils are used for many food and industrial purposes, and the oil quality is associated with its FA composition. Therefore fatty acid composition of edible oils is also dependent on the plant variety, degree of ripening seeds, and climatic conditions. Several studies have documented that temperature is the predominant environmental factor affecting FA composition, especially for unsaturated fatty acids (UFAs) (Varnham, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Jedidi et al.,2020).The results of our study indicate that method of oil extraction also has an influence on the fatty acid composition of the oils.\u003c/p\u003e\u003cp\u003eMezzomo et al. (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) reported that the oil extraction techniques used to obtain high aggregate value compounds from natural products are critical for product quality. The three GNOs showed few variations with either increase or decrease in fatty acid composition from between the hexane extracted crude form to refined form to cold pressed oil. However, the significant variation was observed only with oleic acid (C18:1n9cis) and linoleic acid (C18:2n6cis) content. Abdolshahi et al. (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) outlined that saturated and unsaturated fatty acid profiles of pistachio oil extracted by Soxhlet and maceration method were not different but their content was statistically different. According to Achinewhu and Akpapunam, (\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2005\u003c/span\u003e) refining does not have much effect on fatty acid composition except for some slight inconsistent decrease in saturated and unsaturated fatty acid. Aluyor et al. (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) also reported that refining did not have much effect on fatty acid composition and that there was no undesirable polymerization of oil during the refining process. In contrast to above findings, our study indicates that method of oil extraction does have a mild impact on the essential fatty acid content in the oils. Cold pressed GNO and hexane extracted crude GNO had higher essential fatty acid contentlike linoleic acid (C18:2n6cis), as compared to refined GNO.\u003c/p\u003e\u003cp\u003eOne of the key strengths of this study lies in its controlled experimental design, particularly the use of a single groundnut batch for both cold-pressed and hexane-extracted oils, which allowed for a focused evaluation of how extraction methods influence fatty acid composition while minimizing genotypic variation. Additionally, the integration of a commercially refined oil sample offers practical relevance, reflecting products typically available to consumers.\u003c/p\u003e\u003cp\u003eHowever, the commercially refined groundnut oil (CRGNO) used in this study was obtained as a representative sample from the retail market, and its varietal origin was not disclosed by the manufacturer. As such, while the CRGNO sample offers practical relevance for comparison from a consumer perspective, we acknowledge that its unknown genotype may introduce variability unrelated to the extraction process. This limitation highlights the importance of sourcing oils from known and controlled groundnut varieties in future studies, or alternatively, comparing a range of commercial oils to better account for this variability.\u003c/p\u003e\u003cp\u003eDespite this, the findings reveal subtle but meaningful variations in essential fatty acid content especially linoleic acid,associated with different extraction and refining techniques, suggesting that processing conditions can influence nutritional quality to a modest extent. As such, this study contributes new insight into the nuanced impact of oil processing on fatty acid profiles, an area that has received limited attention in groundnut oil research. Future investigations would benefit from including multiple groundnut varieties with clearly documented genetic backgrounds, as well as a broader spectrum of commercial oils, to more comprehensively assess the interplay between genotype and processing. Collectively, this work augments our understanding of the relationship between extraction method and oil quality, offering beneficial implications for both industry practices, consumer safety and public health nutrition.\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eThe comparative analysis of fatty acid profiles across commercially refined, cold-pressed, and hexane-extracted groundnut oils revealed no statistically significant differences in the overall proportions of total saturated fatty acids, total unsaturated fatty acids, or the unsaturated-to-saturated (U/S) ratio. However, a significant modulation of individual fatty acid components was observed. Specifically, commercially refined groundnut oil exhibited a marked increase in oleic acid (C18:1n9cis) and a decrease in linoleic acid (C18:2n6cis). These results demonstrate that the oil extraction methodology exerts a discernible influence on the retention of essential fatty acids. Notably, cold-pressed and hexane-extracted crude groundnut oils presented a higher linoleic acid (C18:2n6cis) content compared to the refined counterpart. Conversely, the increased oleic acid (C18:1n9cis) levels in the commercially refined oil suggest an enhanced oxidative stability, potentially translating to improved shelf life relative to the cold-pressed and hexane-extracted oils. This study accentuate the critical impact of extraction processes on the final fatty acid composition of groundnut oil, with direct implications for its nutritional value and oxidative stability.\u003c/p\u003e\u003cp\u003eThese findings highlights the subtle yet important ways in which extraction methods shape the nutritional and functional properties of edible oils. The study contributes to the growing body of evidence that, beyond the seed genotype, post-harvest processing steps exert a meaningful influence on oil quality. From a nutritional science perspective, this highlights the trade-offs between stability and essential fatty acid retention when choosing oil processing methods. The insights gained here have practical relevance for both food manufacturers aiming to optimize oil quality for specific culinary uses and for health-conscious consumers seeking oils with superior nutritional profiles. Moreover, this research emphasizes the need for greater transparency in labeling and sourcing, particularly as demand for minimally processed and functional foods continues to rise.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eKBP: Conceptualization, Data curation, Resource Acquisition, Statistical analysis, Investigation, Methodology, Writing original draft. AK.: Conceptualization, Data curation, Methodology,, Supervision, Visualization, Writing original draft. Supervision and validation of results. All authors reviewed the manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eThe authors gratefully acknowledge the Regional Agricultural Research Station, Palem, Professor Jayashankar Telangana State Agricultural University, Nagarkurnool District, Telangana, and the Agricultural Research Station, Kadiri, Acharya N G Ranga Agricultural University, Ananthapur District, Andhra Pradesh, for providing the Kadiri 6 groundnut variety used in this study. We extend our sincere thanks to the MFPI-Quality Control Laboratory, PJTS Agricultural University, Hyderabad, for their support and assistance in the analysis. We also acknowledge the Indian Council of Agricultural Research (ICAR) for their fellowship support.\u003c/p\u003e\u003ch2\u003eData availability\u003c/h2\u003e\u003cp\u003eThe data pertaining to the outcomes of this research can be accessed upon request from the corresponding author.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eShamim A, Nauman K, Iftikhar A, Armghan S, Hafiz ARS (2014) Physicochemical Characteristics, Functional Properties, and Nutritional Benefits of Peanut Oil: A Review. Crit Rev Food Sci Nutr 54(12):1562\u0026ndash;1575\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMaestri D (2024) Groundnut and tree nuts: a comprehensive review on their lipid components, phytochemicals, and nutraceutical properties. 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J King Saud Univ \u0026ndash; Sci 32:2468\u0026ndash;2473\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMezzomo N, Mileo B, Friedrich M, Mart\u0026iacute;nez J, Ferreira S (2010) Supercritical fluid extraction of peach (\u003cem\u003ePrunus persica\u003c/em\u003e) almond oil: Process yield and extract composition. Bioresour Technol 101(14):5622\u0026ndash;5632\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAchinewhu SC, Akpapunam MA (2005) Physical and chemical characteristics of refined vegetable oils from rubber seed and bread fruit. J Plant Food 35:103\u0026ndash;107\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAluyor EO, Aluyor P, Ozigagu CE (2009) Effect of refining on the quality and composition of groundnut oil. Afr J Food Sci 3(8):201\u0026ndash;205\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"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":"food-analytical-methods","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Food Analytical Methods](https://www.springer.com/journal/12161)","snPcode":"12161","submissionUrl":"https://submission.nature.com/new-submission/12161/3","title":"Food Analytical Methods","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Groundnut, Cold-pressed, Hexane extracted oil, Refining, Fatty acids","lastPublishedDoi":"10.21203/rs.3.rs-7142039/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7142039/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eGroundnut (\u003cem\u003eArachis hypogaea\u003c/em\u003e L.) is one of the important oilseed crops in the world, which is utilized for cooking and in confectionery, and is a rich source of valuable antioxidants like vitamin E and plant sterols especially β-sitosterol. In this study, we attempted to study the differences in fatty acid composition in commercially available refined groundnut oil (CRGNO), hexane extracted crude groundnut oil (HCGNO) and cold pressed groundnut oil (CPGNO).The ratio of unsaturated fatty acid to fatty acids was 5.47\u0026thinsp;\u0026plusmn;\u0026thinsp;3.09, 5.91\u0026thinsp;\u0026plusmn;\u0026thinsp;2.96, and 5.34\u0026thinsp;\u0026plusmn;\u0026thinsp;3.11 for CPGNO, CRGNO and HCGNO respectively. Oleic acid (C18:1n9cis) and Linoleic acids (C18:2n6cis) were the major unsaturated fatty acidsand Palmitic acid (C16:0)was the highest saturated fatty acid identified in all the three GNO oils.The total unsaturated fatty acid content was high in all the three oils (86.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23% in CRGNO, 85.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26% in HCGNO and 84.74\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17% in CPGNO).A lower percentage of saturated fatty acids was observed in all the three oils i.e., 15.93\u0026thinsp;\u0026plusmn;\u0026thinsp;5.95%, 15.48\u0026thinsp;\u0026plusmn;\u0026thinsp;6.00% and 14.58\u0026thinsp;\u0026plusmn;\u0026thinsp;5.68% in HCGNO, CPGNO and CRGNO respectively. From the fatty acid analysis of the three oils, it was observed that there was minor variation in the Oleic acid (C18:1n9cis) and Linoleic acids (C18:2n6cis) content and no significant difference in other fatty acids composition among the three samples. Cold pressed and hexane extracted crude GNO had higher essential fatty acid content (linoleic acid (C18:2n6cis) compared to refined GNO, indicating the effect of oil extraction on the fatty acid composition.\u003c/p\u003e","manuscriptTitle":"Effect of Oil Extraction Process on the Fatty Acid Composition of Groundnut Oil","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-29 20:31:59","doi":"10.21203/rs.3.rs-7142039/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-12-20T08:51:21+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-19T16:01:35+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"322931521605439362786067046912993226588","date":"2025-12-17T19:19:50+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-24T09:58:55+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"318050477190533318791256502986042826587","date":"2025-10-15T13:35:44+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"260094201119183908666394833175175400408","date":"2025-10-12T13:21:39+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-08-20T15:32:06+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-07-17T23:49:34+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-07-17T23:48:14+00:00","index":"","fulltext":""},{"type":"submitted","content":"Food Analytical Methods","date":"2025-07-16T16:15:56+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"food-analytical-methods","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Food Analytical Methods](https://www.springer.com/journal/12161)","snPcode":"12161","submissionUrl":"https://submission.nature.com/new-submission/12161/3","title":"Food Analytical Methods","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"7d9c8a17-6afd-403b-8258-cff070ec209d","owner":[],"postedDate":"August 29th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[],"tags":[],"updatedAt":"2025-12-20T08:53:47+00:00","versionOfRecord":[],"versionCreatedAt":"2025-08-29 20:31:59","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7142039","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7142039","identity":"rs-7142039","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}