Antibacterial Study, Phytochemical Analysis and Ft-ir Characterization of Decarboxylated Cashew Nut Shell Liquid (Cnsl) From Cashew Nut Shell

Antibacterial Study, Phytochemical Analysis and Ft-ir Characterization of Decarboxylated Cashew Nut Shell Liquid (Cnsl) From Cashew Nut Shell | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Antibacterial Study, Phytochemical Analysis and Ft-ir Characterization of Decarboxylated Cashew Nut Shell Liquid (Cnsl) From Cashew Nut Shell Victory Omozejele Okodugha, Godwin Aihebhooria Emuokhonun, Joshua Osaretin Onaifo, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6142645/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The cashew nut shell is a viable agriculture waste due to its chemical constituents. The cashew nut shell was macerated with n-hexane for 72-hours to obtain cashew nut shell liquid (CNSL). The CNSL obtained was subjected to decarboxylation reaction through heating at 140 o C for 2 hours. FTIR analysis was carried out on the decarboxylated CNSL which resulted into various peaks: 3362.1cm − 1 indicate medium O-H stretching, alcohol. 2926.0cm − 1 indicate O-H stretching, carboxylic acid. The peak at 1591.6cm − 1 indicate medium C = C stretching, aromatic ring stretch. Phytochemical screening was carried out on the decarboxylated CNSL using standard methods. The phytochemical screening result showed the presence of alkaloids, flavonoids, terpenoids, tannins, saponins, steroids, and phenols. The antibacterial study was done using four bacterial isolates: E.coli, Enterobacter cloacae, Bacillus subtilis and Serratia marsecens. The result showed that decarboxylated CNSL was active against all the bacterial isolates studied at higher concentrations(100% and 75%). Biological sciences/Biochemistry Biological sciences/Biological techniques Biological sciences/Chemical biology Biological sciences/Developmental biology Biological sciences/Microbiology Biological sciences/Molecular biology Biological sciences/Plant sciences Health sciences/Medical research Physical sciences/Chemistry Cashew CNSL decarboxylated CNSL antibacterial Figures Figure 1 Figure 2 Introduction Our day to day survival is plant dependent as they are source of food for the human body needed nutrients. They also provide oxygen needed for human respiration as well as providing raw materials for various industries [ 1 ]. Inaddition, plants help in the provision of shelter, and source of bioactive ingredients that has the potency to fight against human ailments [ 2 ]. The plant kingdom has become the biggest base of essential drugs and there is an increasing awareness of researchers on the importance of medicinal agents found in these plants. The drugs from these medicinal plants are readily available, less toxic, less expensive, safe and effective against various disease causing microorganisms [ 3 ]. Medicinal plants are rich source of antitumor, anti-inflammatory, antimicrobial agents and they serve as therapeutic agents across the globe as a good source of many potent and powerful drugs [ 4 ]. In a bid to control soil erosion in some parts of Nigeria, there was need for afforestation programs. Cashew nut were introduced into Nigeria by the Portuguese in the 15th century for the reduction of the menace caused by erosion. Across the globe, cashew nut is one of the most beneficial nuts in the international trade market [5, 6, 7]. The cashew tree is evergreen which is responsible for the production of the cashew nut. The thickness of the cashew nut is about 1/8 inches. Inside the nut is a soft honey comb structure containing a dark brown viscous liquid called the cashew nut shell liquid [ 8 ]. The nut is filled with cashew nut shell liquid (CNSL) and this CNSL is primarily composed of anacardic acid, cardanol, cardol and 2-methyl cardol. These four components differ in their side chain. CNSL is known to contain 70% of anacardic acid and when heated about 90% of cardanol is formed [9, 10, 11]. CNSL is known to possess antioxidant, antitumor, anti-inflammatory properties and show biological activity against Alzheimer's disease [ 11 , 12 , 13 ]. Materials and Methods Collection and identification of plant material Fresh cashew nut shell were collected from a farm land at Ujemen, Esan west local goverment Area of Edo state. The plant material was identified by the Department of Plant Biology and Biotechnology, University of Benin, Benin City. The plant material was air dried and reduced into smaller masses for further analysis. Extraction of CNSL 150g of the cashew nut shell was weighed using electronic weighing balance and soaked with 500ml of n-hexane using 1000ml conical flask. The mixture was allowed to stand for 72-hours followed by intermittent stirring to enhance extraction. The mixtures was filtered and the filterate was concentrated using Rotary evaporator. Decarboxylation of CNSL The extracted CNSL was heated at 140 o C for 2 hours [14]. Phytochemical screening of decarboxylated CNSL This was done using standard methods Antibacterial study of decarboxylated CNSL Preparation of various concentration of Decarboxylated CNSL D ecarboxylated CNSL, was disclosed in separate tween 80 to obtain varying concentrations (100%, 75%, 50% and 25%). 10 ml was used as the standard volume. 100% concentration was attained by pipetting 10 ml of decarboxylated CNSL into sterile universal bottle containing 10ml of tween 80. 75% concentration was obtained by pipetting 7.5ml of decarboxylated CNSL into 7.5ml of tween 80. 50% concentration was obtained by dissolving 5ml of decarboxylated CNSL into 5ml of tween 80. 25% concentration was obtained by dissolving 2.5ml of decarboxylated CNSL into 2.5ml of tween 80. Activation of microbial strains and standardization of inoculum The organisms used for this study were subcultured into Mueller Hinton medium plates and incubated for 24 hours at 37 o C. The agar plates were stored at 4 o C until required. Determination of Minimum Inhibitory Concentration (MIC) A drop of bacterial suspension, which had been previously diluted was aseptically added to molten nutrient agar and left to sterilize for a duration of 24 hours, the plates were incubated at 37℃. For every test organism, MIC was determined to be the lowest concentration that hindered observable growth. Determination of Minimum Bactericidal Concentration (MBC) To plate out the contents of the lowest MIC plate in each region of the plate, nutrient agar plates were separated into separate pieces and labelled with the corresponding concentration on the base of the plates. The MBC were measured after the plates were aerobically incubated at 37℃ for 24 hours. MBC was defined as the lowest concentration of MIC tubes showing no signs of visible bacterial growth. Result Table 1 Phytochemical analysis of decarboxylated CNSL Phytochemical screening Result Alkaloids + Flavonoids + Terpenoids + Steriods + Saponins + Phenols ++ Tannins + Keys: + = present, ++ = highly present , Table 3 Minimum inhibitory concentration (MIC) and Minimum bactericidal concentration (MBC) of Decarboxylated CNSL Minimum Inhibitory Concentration Minimum Bactericidal concentration E. coli 57 0 Enterobacter cloacae 55 0 Bacillus subtilis 55 0 Serratia marsecens 55 0 Discussion In Fig. 1 , the peak of absorption that appeared at 3362.1 cm−1 indicate the presence of -OH group of phenolic compounds. The peak at 1722.0 cm−1 indicate weak, C-H bending of aromatic compounds. At 1591.6 cm−1 we observed medium, C = C stretching indicating aromatic ring stretch. The peak at 1457.4cm−1 indicates C = C-C stretch, aromatic ring while C-H, 1,3- disubstituted (meta) was observed at 779.0 cm−1. Figure 2 indicate the MIC and MBC of decarboxylated CNSL against bacterial isolates. The MIC values ranges from 55 to 57 for all the bacterial isolates used for this study. However the MBC was zero for all the bacterial isolates studied. The phytochemical analysis result in Table 1 , shows the presence of alkaloids, flavonoids, terpenoids, tannins, saponins, steroids and phenols in decarboxylated CNSL. Table 2 shows that decarboxylated CNSL was more effective against all the bacterial isolates studied at 100% zone of inhibition against all the bacterial isolates ( E.coli, Enterobacter cloacae, Bacillus subtilis, Serratia marsecens) studied. At 100% concentration, the activity of decarboxylated CNSL ranges from 8 to 12. At 75% concentration, decarboxylated CNSL was active against E.coli, Enterobacter cloacae and Bacillus subtilis. It was however inactive against Serratia marsecens. At 50% concentration, decarboxylated CNSL was found to be active against Bacillus subtilis only. It was inactive against E.coli, Enterobacter cloacae and Serratia marsecens. At 25% concentration, decarboxylated CNSL was inactive against all the bacterial isolates studied. The activity of decarboxylated CNSL against all the studied bacterial isolates was however lower than that of the control drug (ciprofloxacin). Table 3 reveal the MIC and MBC values of decarboxylated CNSL. The MIC value ranges from 55–57. However, the MBC value was found to be zero. Conclusion The impact of decarboxylated CNSL on bacterial isolates used for this study at 100% and 75% concentrations shows that decarboxylated CNSL has basic medicinal agents. Further research can be carried out to harness these biological active ingredients present in decarboxylated CNSL. Declarations Acknowledgement The authors want to thank the laboratory staff of the Department of Chemistry both at Ambrose Alli University and University of Benin for their cooperation throughout this research work. Clinical trial Not applicable Ethics and Consent to participate declaration Not applicable Consent to Publish declaration Not applicable Competing Interest declaration There are no competing interest Funding Not applicable Data availability All data generated or analysed during this study are included in this published article. References Abulude FO, Ogunkoya MO, Akinjagunla YS. Phytochemical screening of leaves and stem of Cashew tree (Anacardium occidentate). Environmental Agricultural and Food Chemistry. 2010;9:815-819. Tipu MS, Akhtar MI, Raja ML. New dimension of medicinal plant as animal feed.Pakistan Journal of Botany. 2006;26(3):144-148. Cathrine L, Prabavathi N. Preliminary phytochemical analysis and antibacterial activity of leaf extracts of Vitex leucoxylon L. F. International Journal of Current Pharmaceutical Research. 2011;3(2):71-73. Srivastava J, Lambert J, Vietmeyer N. Medicinal plants: An expanding role in development. World Bank Technical. 1996;320. Asogwa EU, Hammed LA, Ndubuaku TCN. Integrated production and protection practices of cashew, Anacardium occidentale in Nigeria. Afr J Biotechnol. 2008; 7(25):4868-4873. Ohler JG. Cashew growing. Department of Agricultural Resources.Tropical Institute. 1979; 7(4):260-271. Pinto AMB, Santos TMP, Caceres CA, Lima JR, Ito EN, Azeredo H MC. Starch-cashew tree gum nano-composite films and their application for coating cashew nuts.LWT -Food Sci Technol (Campinas).2015; 62:549-554. Tejas G, Mayank P and Bharat K.D. (2012). Studies on effect of various solvents on extraction of cashew nut shell liquid (CNSL) and isolation of major phenolic constituents from extracted CNSL. Journal of Natural Product and Plant Resource. 2(1):135. Lomonaco D, Mele G and Mazzetto S.E. (2017). Cashew nut shell liquid(CNSL): From an agro-industrial waste to a sustainable alternative to petrochemical resources. Springer, Cham. 19:38. Mubofu E.B and Mgaya J.E. (2018). Chemical valorization of cashew nut shell waste. Top. Curr. Chem. 376: 8. Lemes L.F.N, De Andrade R.G, A.S. Oliveira A.S, Da Silva F.M.R, De Castro C.G, Da Silva B.M, Romeiro L.A.S. (2016). Cardanol-derived AChE inhibitors: Towards the development of dual binding derivatives for Alzheimer’s disease. European Journal of Medicinal Chemistry, 108: 687-700. Huang H, Hua N. Liu X, Li S. Liu X, Chen J. Liu. (2014). Anacardic acid induces cell apoptosis associated with induction of ATF4-dependent endoplasmic reticulum stress. Toxicology Letters, 228 (3): 170-178. Andrade T.D.J.A.D.S, Araújo B.Q, Citó A.M.D.G.L, Da Silva J, Saffi J, Richter M.F, Ferraz A.D.B.F.? (2011). Antioxidant properties and chemical composition of technical Cashew Nut Shell Liquid (tCNSL). Food Chemistry, 126 (3): 1044-1048. Njuku, F.W. Mwangi, P.M. and Thiong'o, G. T. (2014). Evaluation of cardanol acetate as a reactive diluent for alkyd coating. International. Journal of Advanced Research. 2: 930. Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6142645","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":428242859,"identity":"4abe6961-9470-4267-96bb-75512b3e018b","order_by":0,"name":"Victory Omozejele 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04:53:20","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6142645/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6142645/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":78671500,"identity":"42488539-9fc0-45f4-8a07-a863ba4a9a92","added_by":"auto","created_at":"2025-03-17 12:40:32","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":231012,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR spectrum of decarboxylated CNSL\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6142645/v1/5b5c5d3553d57b67cc3be3af.png"},{"id":78670745,"identity":"94440288-198b-4917-bb9a-f69e1d5c2348","added_by":"auto","created_at":"2025-03-17 12:32:32","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":79176,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMIC and MBC of decarboxylated CNSL\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6142645/v1/30d4d113b6ecfde98d3963ca.png"},{"id":96167443,"identity":"00339832-e8a2-42ea-9750-f54cfa2d2b7c","added_by":"auto","created_at":"2025-11-18 09:54:45","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1005617,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6142645/v1/bc7380a8-14dd-4d08-990c-a51a81daf980.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eAntibacterial Study, Phytochemical Analysis and Ft-ir Characterization of Decarboxylated Cashew Nut Shell Liquid (Cnsl) From Cashew Nut Shell\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eOur day to day survival is plant dependent as they are source of food for the human body needed nutrients. They also provide oxygen needed for human respiration as well as providing raw materials for various industries [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Inaddition, plants help in the provision of shelter, and source of bioactive ingredients that has the potency to fight against human ailments [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe plant kingdom has become the biggest base of essential drugs and there is an increasing awareness of researchers on the importance of medicinal agents found in these plants. The drugs from these medicinal plants are readily available, less toxic, less expensive, safe and effective against various disease causing microorganisms [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Medicinal plants are rich source of antitumor, anti-inflammatory, antimicrobial agents and they serve as therapeutic agents across the globe as a good source of many potent and powerful drugs [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn a bid to control soil erosion in some parts of Nigeria, there was need for afforestation programs. Cashew nut were introduced into Nigeria by the Portuguese in the 15th century for the reduction of the menace caused by erosion. Across the globe, cashew nut is one of the most beneficial nuts in the international trade market [5, 6, 7].\u003c/p\u003e \u003cp\u003eThe cashew tree is evergreen which is responsible for the production of the cashew nut. The thickness of the cashew nut is about 1/8 inches. Inside the nut is a soft honey comb structure containing a dark brown viscous liquid called the cashew nut shell liquid [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe nut is filled with cashew nut shell liquid (CNSL) and this CNSL is primarily composed of anacardic acid, cardanol, cardol and 2-methyl cardol. These four components differ in their side chain. CNSL is known to contain 70% of anacardic acid and when heated about 90% of cardanol is formed [9, 10, 11]. CNSL is known to possess antioxidant, antitumor, anti-inflammatory properties and show biological activity against Alzheimer's disease [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eCollection and identification of plant material\u003c/h2\u003e \u003cp\u003eFresh cashew nut shell were collected from a farm land at Ujemen, Esan west local goverment Area of Edo state. The plant material was identified by the Department of Plant Biology and Biotechnology, University of Benin, Benin City. The plant material was air dried and reduced into smaller masses for further analysis.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eExtraction of CNSL\u003c/h3\u003e\n \u003cp\u003e150g of the cashew nut shell was weighed using electronic weighing balance and soaked with 500ml of n-hexane using 1000ml conical flask. The mixture was allowed to stand for 72-hours followed by intermittent stirring to enhance extraction. The mixtures was filtered and the filterate was concentrated using Rotary evaporator.\u003c/p\u003e \u003cp\u003e \u003cb\u003eDecarboxylation of CNSL\u003c/b\u003e \u003c/p\u003e \u003cp\u003eThe extracted CNSL was heated at 140\u003csup\u003eo\u003c/sup\u003eC for 2 hours [14].\u003c/p\u003e \u003cp\u003e \u003cb\u003ePhytochemical screening of decarboxylated CNSL\u003c/b\u003e \u003c/p\u003e \u003cp\u003eThis was done using standard methods\u003c/p\u003e \u003cp\u003e \u003cb\u003eAntibacterial study of decarboxylated CNSL\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003ePreparation of various concentration of Decarboxylated CNSL\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eD\u003c/b\u003eecarboxylated CNSL, was disclosed in separate tween 80 to obtain varying concentrations (100%, 75%, 50% and 25%). 10 ml was used as the standard volume. 100% concentration was attained by pipetting 10 ml of decarboxylated CNSL into sterile universal bottle containing 10ml of tween 80. 75% concentration was obtained by pipetting 7.5ml of decarboxylated CNSL into 7.5ml of tween 80. 50% concentration was obtained by dissolving 5ml of decarboxylated CNSL into 5ml of tween 80. 25% concentration was obtained by dissolving 2.5ml of decarboxylated CNSL into 2.5ml of tween 80.\u003c/p\u003e \u003cp\u003e \u003cb\u003eActivation of microbial strains and standardization of inoculum\u003c/b\u003e \u003c/p\u003e \u003cp\u003eThe organisms used for this study were subcultured into Mueller Hinton medium plates and incubated for 24 hours at 37 \u003csup\u003eo\u003c/sup\u003eC. The agar plates were stored at 4 \u003csup\u003eo\u003c/sup\u003eC until required.\u003c/p\u003e \u003cp\u003e \u003cb\u003eDetermination of Minimum Inhibitory Concentration (MIC)\u003c/b\u003e \u003c/p\u003e \u003cp\u003eA drop of bacterial suspension, which had been previously diluted was aseptically added to molten nutrient agar and left to sterilize for a duration of 24 hours, the plates were incubated at 37℃. For every test organism, MIC was determined to be the lowest concentration that hindered observable growth.\u003c/p\u003e \u003cp\u003e \u003cb\u003eDetermination of Minimum Bactericidal Concentration (MBC)\u003c/b\u003e \u003c/p\u003e \u003cp\u003eTo plate out the contents of the lowest MIC plate in each region of the plate, nutrient agar plates were separated into separate pieces and labelled with the corresponding concentration on the base of the plates. The MBC were measured after the plates were aerobically incubated at 37℃ for 24 hours. MBC was defined as the lowest concentration of MIC tubes showing no signs of visible bacterial growth.\u003c/p\u003e"},{"header":"Result","content":"\u003cdiv\u003e\n \u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv\u003eTable 1\u003c/div\u003e\n \u003cdiv\u003e\n \u003cp\u003ePhytochemical analysis of decarboxylated CNSL\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePhytochemical screening\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eResult\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eAlkaloids\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eFlavonoids\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eTerpenoids\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e+\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eSteriods\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e+\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eSaponins\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e+\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003ePhenols\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e++\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eTannins\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e+\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\"\u003e\u003cstrong\u003eKeys: + = present, ++ = highly present\u003c/strong\u003e,\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003cdiv align=\"char\"\u003e\u003cimg src=\"https://myfiles.space/user_files/122228_c8a1650c59388082/122228_custom_files/img1742186956.png\"\u003e\u003cbr\u003e\u003c/div\u003e\n \u003cdiv align=\"char\"\u003e\u003cbr\u003e\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv\u003e\n \u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv\u003eTable 3\u003c/div\u003e\n \u003cdiv\u003e\n \u003cp\u003eMinimum inhibitory concentration (MIC) and Minimum bactericidal concentration (MBC) of Decarboxylated CNSL\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMinimum Inhibitory Concentration\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMinimum Bactericidal concentration\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eE. coli\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eEnterobacter cloacae\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBacillus subtilis\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSerratia marsecens\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, the peak of absorption that appeared at 3362.1 cm−1 indicate the presence of -OH group of phenolic compounds. The peak at 1722.0 cm−1 indicate weak, C-H bending of aromatic compounds. At 1591.6 cm−1 we observed medium, C = C stretching indicating aromatic ring stretch. The peak at 1457.4cm−1 indicates C = C-C stretch, aromatic ring while C-H, 1,3- disubstituted (meta) was observed at 779.0 cm−1. Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e indicate the MIC and MBC of decarboxylated CNSL against bacterial isolates. The MIC values ranges from 55 to 57 for all the bacterial isolates used for this study. However the MBC was zero for all the bacterial isolates studied. The phytochemical analysis result in Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, shows the presence of alkaloids, flavonoids, terpenoids, tannins, saponins, steroids and phenols in decarboxylated CNSL. Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e shows that decarboxylated CNSL was more effective against all the bacterial isolates studied at 100% zone of inhibition against all the bacterial isolates ( E.coli, Enterobacter cloacae, Bacillus subtilis, Serratia marsecens) studied. At 100% concentration, the activity of decarboxylated CNSL ranges from 8 to 12. At 75% concentration, decarboxylated CNSL was active against E.coli, Enterobacter cloacae and Bacillus subtilis. It was however inactive against Serratia marsecens. At 50% concentration, decarboxylated CNSL was found to be active against Bacillus subtilis only. It was inactive against E.coli, Enterobacter cloacae and Serratia marsecens. At 25% concentration, decarboxylated CNSL was inactive against all the bacterial isolates studied. The activity of decarboxylated CNSL against all the studied bacterial isolates was however lower than that of the control drug (ciprofloxacin). Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e reveal the MIC and MBC values of decarboxylated CNSL. The MIC value ranges from 55–57. However, the MBC value was found to be zero.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe impact of decarboxylated CNSL on bacterial isolates used for this study at 100% and 75% concentrations shows that decarboxylated CNSL has basic medicinal agents. Further research can be carried out to harness these biological active ingredients present in decarboxylated CNSL.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors want to thank the laboratory staff of the Department of Chemistry both at Ambrose Alli University and University of Benin for their cooperation throughout this research work.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical trial\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics and Consent to participate declaration\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Publish declaration\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interest declaration\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere are no competing interest\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eNot applicable\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analysed during this study are included in this published article.\u003c/p\u003e\n"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbulude FO, Ogunkoya MO, Akinjagunla YS. Phytochemical screening of leaves and stem of Cashew tree (Anacardium occidentate). Environmental Agricultural and Food Chemistry. 2010;9:815-819.\u003c/li\u003e\n\u003cli\u003eTipu MS, Akhtar MI, Raja ML. New dimension of medicinal plant as animal feed.Pakistan Journal of Botany. 2006;26(3):144-148.\u003c/li\u003e\n\u003cli\u003eCathrine L, Prabavathi N. Preliminary phytochemical analysis and antibacterial activity of leaf extracts of Vitex leucoxylon L. F. International Journal of Current Pharmaceutical Research. 2011;3(2):71-73.\u003c/li\u003e\n\u003cli\u003eSrivastava J, Lambert J, Vietmeyer N. Medicinal plants: An expanding role in development. World Bank Technical. 1996;320.\u003c/li\u003e\n\u003cli\u003eAsogwa EU, Hammed LA, Ndubuaku TCN. Integrated production and protection practices of cashew, Anacardium occidentale in Nigeria. Afr J Biotechnol. 2008; 7(25):4868-4873. \u003c/li\u003e\n\u003cli\u003eOhler JG. Cashew growing. Department of Agricultural Resources.Tropical Institute. 1979; 7(4):260-271.\u003c/li\u003e\n\u003cli\u003ePinto AMB, Santos TMP, Caceres CA, Lima JR, Ito EN, Azeredo H MC. Starch-cashew tree gum nano-composite films and their application for coating cashew nuts.LWT -Food Sci Technol (Campinas).2015; 62:549-554.\u003c/li\u003e\n\u003cli\u003eTejas G, Mayank P and Bharat K.D. (2012). Studies on effect of various solvents on extraction of cashew nut shell liquid (CNSL) and isolation of major phenolic constituents from extracted CNSL. Journal of Natural Product and Plant Resource. 2(1):135.\u003c/li\u003e\n\u003cli\u003eLomonaco D, Mele G and Mazzetto S.E. (2017). Cashew nut shell liquid(CNSL): From an agro-industrial waste to a sustainable alternative to petrochemical resources. Springer, Cham. 19:38.\u003c/li\u003e\n\u003cli\u003eMubofu E.B and Mgaya J.E. (2018). Chemical valorization of cashew nut shell waste. Top. Curr. Chem. 376: 8.\u003c/li\u003e\n\u003cli\u003eLemes L.F.N, De Andrade R.G, A.S. Oliveira A.S, Da Silva F.M.R, De Castro C.G, Da Silva B.M, Romeiro L.A.S. (2016). Cardanol-derived AChE inhibitors: Towards the development of dual binding derivatives for Alzheimer\u0026rsquo;s disease. European Journal of Medicinal Chemistry, 108: 687-700.\u003c/li\u003e\n\u003cli\u003eHuang H, Hua N. Liu X, Li S. Liu X, Chen J. Liu. (2014). Anacardic acid induces cell apoptosis associated with induction of ATF4-dependent endoplasmic reticulum stress. Toxicology Letters, 228 (3): 170-178.\u003c/li\u003e\n\u003cli\u003eAndrade T.D.J.A.D.S, Ara\u0026uacute;jo B.Q, Cit\u0026oacute; A.M.D.G.L, Da Silva J, Saffi J, Richter M.F, Ferraz A.D.B.F.? (2011). Antioxidant properties and chemical composition of technical Cashew Nut Shell Liquid (tCNSL). Food Chemistry, 126 (3): 1044-1048.\u003c/li\u003e\n\u003cli\u003eNjuku, F.W. Mwangi, P.M. and Thiong\u0026apos;o, G. T. (2014). Evaluation of cardanol acetate as a reactive diluent for alkyd coating. International. Journal of Advanced Research. 2: 930.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Cashew, CNSL, decarboxylated CNSL, antibacterial","lastPublishedDoi":"10.21203/rs.3.rs-6142645/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6142645/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe cashew nut shell is a viable agriculture waste due to its chemical constituents. The cashew nut shell was macerated with n-hexane for 72-hours to obtain cashew nut shell liquid (CNSL). The CNSL obtained was subjected to decarboxylation reaction through heating at 140\u003csup\u003eo\u003c/sup\u003eC for 2 hours. FTIR analysis was carried out on the decarboxylated CNSL which resulted into various peaks: 3362.1cm\u0026thinsp;\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e indicate medium O-H stretching, alcohol. 2926.0cm\u0026thinsp;\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e indicate O-H stretching, carboxylic acid. The peak at 1591.6cm\u0026thinsp;\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e indicate medium C\u0026thinsp;=\u0026thinsp;C stretching, aromatic ring stretch. Phytochemical screening was carried out on the decarboxylated CNSL using standard methods. The phytochemical screening result showed the presence of alkaloids, flavonoids, terpenoids, tannins, saponins, steroids, and phenols. The antibacterial study was done using four bacterial isolates: E.coli, Enterobacter cloacae, Bacillus subtilis and Serratia marsecens. The result showed that decarboxylated CNSL was active against all the bacterial isolates studied at higher concentrations(100% and 75%).\u003c/p\u003e","manuscriptTitle":"Antibacterial Study, Phytochemical Analysis and Ft-ir Characterization of Decarboxylated Cashew Nut Shell Liquid (Cnsl) From Cashew Nut Shell","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-17 12:32:28","doi":"10.21203/rs.3.rs-6142645/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"926c95be-9601-4f72-981a-0ea13bde5f7b","owner":[],"postedDate":"March 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":45755288,"name":"Biological sciences/Biochemistry"},{"id":45755289,"name":"Biological sciences/Biological techniques"},{"id":45755290,"name":"Biological sciences/Chemical biology"},{"id":45755291,"name":"Biological sciences/Developmental biology"},{"id":45755292,"name":"Biological sciences/Microbiology"},{"id":45755293,"name":"Biological sciences/Molecular biology"},{"id":45755294,"name":"Biological sciences/Plant sciences"},{"id":45755295,"name":"Health sciences/Medical research"},{"id":45755296,"name":"Physical sciences/Chemistry"}],"tags":[],"updatedAt":"2025-11-18T09:53:42+00:00","versionOfRecord":[],"versionCreatedAt":"2025-03-17 12:32:28","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6142645","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6142645","identity":"rs-6142645","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}