Extraction and Phytochemical Screening of Secondary Metabolites from Leonotis Ocymifolia (Bokolu) plant on areal part

Extraction and Phytochemical Screening of Secondary Metabolites from Leonotis Ocymifolia (Bokolu) plant on areal part | 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 Extraction and Phytochemical Screening of Secondary Metabolites from Leonotis Ocymifolia (Bokolu) plant on areal part Defar Bekele Adugna This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6429306/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 Nowadays, the uses of medicinal plants are more diverse to prevent human health care, in which medicinal plants are powerful inhibition of microorganisms cause disease. The study is conducted to assess extraction, phytochemical screening and methods of extracting secondary metabolites of a traditionally used medicinal plant, Leonotis ocymifolia. The plant has been extracted using 80% aqueous ethanol by maceration technique, and 6.0 g of grey ethanol crude were obtained. The crude ethanol suspended in water and partitioned within different polarity of solvents (petroleum ether, ethylene acetate, and n-buthanol), respectively. During partitioning, 2.15 g of ethylene acetate were collected after concentrated using a rotary evaporator and few grams of petroleum ether were obtained. The study revealed phytochemical components like alkaloids, flavonoids, phenols, tannins, glycosides, terpeniods and steroids. Physical sciences/Chemistry Physical sciences/Materials science Physical sciences/Nanoscience and technology phytochemical extraction partition isolation Figures Figure 1 Figure 2 Figure 3 Introduction Medicinal plants are now more focused than ever because they have the capability of producing many undeniable benefits to people, especially in the line of medicine and pharmacological study. The medicinal power of these plants lies in phytochemical constituents that cause definite pharmacological actions on growth of pathogens 1 . Phytochemicals are natural compounds produced in plants such as alkaloids, glycosides, flavonoids, tannins, steroids, terpenoids and phenols are of secondary metabolites of plant that serves as defensives agents of microorganism’s 2 . Around worldwide, including Ethiopia traditional medicinal plant is more applicable to treat disease and infection starting from ancient time. The use of traditional medicine in Ethiopia is based on indigenous knowledge and has long been practiced in the treatment of various diseases 3 . For centuries, people have relied heavily on traditional medicine to treat a variety of physical and mental ailments. Approximately 80% of people in the country still rely on traditional medicine as their primary healthcare system 4 , and more than 95% consists of traditional herbal medicines. In Ethiopia, many plant species used as a medicinal plant and Leonotis ocymifolia is typically used plant almost all part of population of Ethiopia 5 . Leonotis ocymifolia is minaret flower (flowering plant), shrub and has along white corolla covered by orange rufous hair which is belong to Lamiaceae family 6 . In Ethiopia, it is known as Bokolu, and it is good traditional medicinal plant for many diseases. The plant is reasonably drought-resistant and wind tolerant. Unlike the similar Leonotus leonuris, in the ocymifolia, the tubular flowers are bolder and larger 7 . Leonotis ocymifolia is folk medicine which is used to treat mich and other belief in Ethiopia 2 , 8 . In society, the plant is used for the diarrheal, headache, cold (influenza), cattle doweling, stomach worm and malaria in some part of Ethiopia specially oromia region[Ethiomedicine]. The main objective of this study was to investigate in vitro isolation of secondary metabolites based on different polarity of solvent and phytochemical screening of secondary metabolites. Results and Discussion Extraction yields 6.0 g of grey crude was produced from the maceration of 100g of L. ocymifolia plant powder. The process of partitioning the ethanol crude involved suspending (dissolving) the crude extracted in an aqueous solution, starting with the least polar solvent (petroleum ether), increasing the polarity of the solvents, and generating a separating layer due to the immiscibility of the two solvents. Following the partitioning of the ethanol crude into ethylene acetate, and n-buthanol crude, respectively, 2.15 g of yellowish ethylene acetate crude and 1.80 g of n-buthanol crude were ultimately produced. However, measuring petroleum ether is extremely difficult. Partition of Petroleum ether Ethanol crude extracts dissolve in aqueous solution and are partitioned within petroleum ether. Due to the immiscibility of the two solvents, a phase boundary is formed. The upper layer (grey color) suspended over the aqueous solution (Figure 1) is petroleum ether crude solution and is carefully separated into a different container. The petroleum ether solution is concentrated by exposure at room temperature, and very few grams of crude were obtained. Partition of Ethylene acetate Ethylene acetate also partitioned within the aqueous ethanol solution, and phase boundary is formed in which the yellowish solution floated over aqueous. The upper layer (yellowish) solution (Figure 2) is ethylene acetate, and the two layers separated. The ethylene crude solution is concentrated, and 2.15g of crude collected. In this partition, relatively high mass proportion of secondary metabolites revealed. Partition of n-buthanol Finally, the aqueous ethanol partitioned within n-butanol (Figure 3), and 1.80 g of crude collected. The remaining aqueous solution discarded because of primary metabolites. Ethanol solvents can extracts from least polar compound to very more polar (primary metabolites). Therefore, partitioning is used to remove these primary metabolites from crude extract and narrow down the range of secondary metabolites based on their polarities. Phytochemical Screening Phytochemical screening of main secondary metabolites such as alkaloids, flavonoids, steroids, saponins, tannins, terpenoids, and phenols was studied on petroleum ether, ethylene acetate, and n-butanol crude partitions. In the ethylene acetate partition, terpenoids, phenols, flavonoids, and steroids were present, whereas phenols, glycosides, and tannins were present in the n-butanol partition (Table 1). Table1: Table of Phytochemical Screening s/no components petroleum ether ethylene acetate n- buthanol 1 alkaloids + - - 2 flavonoids - + - 3 phenols - + + 4 glycosides - - + 5 terpenoids - + - 6 saponins - - + 7 tannins - - - 8 steroids - + - In general, the extraction of the aerial parts of Leonotis ocymifolia in this study revealed secondary metabolites like alkaloids, flavonoids, phenols, glycosides, terpinoids, tannins, and steroids and the result similar within 3 . In contrast, saponins are not present in any of the crude oil partitions, and components are indicated by the "+" sign, while "-" denotes the absence of a component (Table 1). Table 1 show that the sugar component (primary metabolites) remained in the aqueous solution while nearly all of the secondary metabolites were detected in both solvents (ethyl acetate and n-butanol partition). The reason for this is that highly polar chemicals are attracted to water as a solvent. Material and methodology Plant collection In December 2024, the mature and healthy Leonotis ocymifolia plants were gathered from a family garden in Goba woreda. Located in the Bale zone of the Oromia region, southeast of Addis Ababa, Ethiopia, lies the Goba woreda. The plant was authenticated by Botanist Reta Regassa, and a voucher specimen with the number BD00130/24 was deposited at the HU Herbarium, Hawassa University, for future reference. A sufficient number of plant sections will be collected, and the newly collected plants will undergo three rounds of washing in the laboratory, first with tap water to get rid of dust and then with distilled water to rinse. Next, after being cut with a knife into small pieces, it dries in the shade. Chemical and Material Chemicals: distilled water, ethanol (80%), ethyl acetate (AR), NaOH ,silica gel (mesh 200-400), petroleum ether(AR), naphthol, iodine, KI, HCl(37%), chloroform(AR), H 2 SO 4 (98%), FeCl 3 , methanol (80%). Material: Erlenmeyer flask, water bath shaker (SHZ -82), rotary evaporator (RE100-S), vacuum pump, column chromatography, TLC, dropper, spatula, electronic balance, Buchner funnel, watch man filter paper, iron stand up, electrical drier, Knife, separating funnel. Extraction The plant's dried sections were ground using an electrical mill, and 100g of finely ground Leonotis ocymifolia powder was soaked in 80% aqueous ethanol in 1:8 ratios for three days. The plant was then shaken for about 36 hours at 200°C using a water bath shaker (SHZ-82). Then watch man filter paper was used to separate the liquid. Grey ethanol crude was obtained by using a rotary evaporator (RE100-S) to concentrate the filtered liquid. Ultimately, ethanol crude was divided into petroleum ether and suspended in distilled water using a separating funnel. The two components were then carefully separated. Subsequently, the remaining aqueous solution was cautiously separated into separate containers after being gradually divided into ethylene acetate and n-buthanol. Ultimately, a rotary evaporator (RE100-S) was used to concentrate each partitioned solution independently. Phytochemicals screening Petroleum ether crude, ethylene acetate, and n-butanol solutions were used to test the phytochemical components. Using standard procedure, this is done to determine whether secondary metabolites are present or absent 9 , 10 . Test for alkaloids Few drops of Mayer’s reagent were added to 1ml of extract. A yellowish or white precipitate was formed, indicating the presence of alkaloids 10 . Test for flavonoids Two to three drops of sodium hydroxide were added to 2 mL of extract. Initially, a deep yellow color appeared but it gradually became colorless by adding few drops of dilute HCL, indicating that flavonoids were present 10 . Test for phenols Two milliliters of 5% neutral ferric chloride solution were added to 1 ml of extract, the dark blue coloring indicating the presence of phenolic compounds 10 . Test for glycosides Few drops of Molisch’s reagent will be added to 2 ml of extract and will be shaken well. 2ml of conc. H 2 SO 4 will be added on the sides of the test tube. A reddish violet ring appeared at the junction of two layers immediately indicated 10 . Test for terpenoids About 5 mL of extract will be dissolved and mixed with 2 ml of chloroform, then 3 mL of concentrated H 2 SO4 were added and reddish brown coloration at the interface wills confirmed the presence of terpenoides 10 . Test for tannins 0.1% FeCl 3 solution added to few ml of extract and the blue, blue-black, green or blue-green color which confirmed the presence of tannins 10 . Test for sponins A drop of Na 2 CO 3 solution was added to 5 ml of extract in a test tube. After vigorous shaking, it was left to rest for five minutes. Foam formation indicated the presence of saponins 10 . Test for steroids 2 ml of chloroform and concentrated H 2 SO 4 were added with the 5 ml aqueous plant crude extract. In the lower chloroform layer red color appeared that indicated the presence of steroids 10 . Conclusion According to the results, the plant is rich within secondary metabolites and further experiments are required to comprehensively, characterized bioactive component and explore their antimicrobial activities. Declarations Data availability The data and material used to support the findings of this study are available main body text and are additional available from the corresponding author upon request. Acknowledgements The author is thankful to Madda Walabu University, Department of Chemistry, for providing laboratory facilities and chemical support during laboratory analysis. The author gratefully thanks Mr. Yohannes Shuka for supporting publication process. Author contributions Defar Bekele: conceptualization, writing (original draft), Conceived and designed the experiments methods; performed the experiments; Analyzed and interpreted the data; Contributed reagents, materials, analysis tools or data; Validation, Writing (review and editing), preparing figures, collecting data, validation investigation, and Formal analysis. Funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for profit sectors. Competing interests The author declares no competing interests. References Yadav, R. & Agarwala, M. Phytochemical analysis of some medicinal plants. J. Phytol. 3 , 10–14 (2011). Agidew, M. G. Phytochemical analysis of some selected traditional medicinal plants in Ethiopia. Bull. Natl. Res. Cent. 46 , (2022). Ayele, T. M., Abebe, E. C. & Kassie, A. B. Investigation of antibacterial and anti-diarrhoeal activities of 80% methanol leaf and fruit extract of leonotis ocymifolia (Burm. f) iwarsson (lamiaceae). Journal of Experimental Pharmacology vol. 13 613–626 at https://doi.org/10.2147/JEP.S319981 (2021). Dubale, S., Kebebe, D., Zeynudin, A., Abdissa, N. & Suleman, S. Phytochemical Screening and Antimicrobial Activity Evaluation of Selected Medicinal Plants in Ethiopia. J. Exp. Pharmacol. 15 , 51–62 (2023). Asteraye, Y. ADDIS ABABA UNIVERSITY OFFICE OF RESEARCH AND GRADUATE PROGRAMS PHYTOCHEMICAL INVESTIGATION ON THE LEAVES OF Leonotis Ocymifolia . (2006). Teklu, T., Engidawork, E., Nedi, T., Teklehaymanot, T. & Gebremeskel, L. Evaluation of the Antimalarial Activity of the Hydroalcoholic Extract of Leaf of Leonotis ocymifolia (Burm. f.) Iwarsson (Lamiaceae) against Plasmodium berghei in Mice. Evidence-based Complement. Altern. Med. 2020 , (2020). A Review on Medicinal Plants Against Some Human Pathogenic Bacteria (E. coli, S.dysentery, S.typhi, P.aeruginosa and S. aureus) in South East Ethiopia. J. Biol. Agric. Healthc. (2019) doi:10.7176/jbah/9-19-03. Talema, A. Review of Phytochemical Analysis of Selected Traditional Medicinal Plants in Ethiopia. Int. J. Homeopath. Nat. Med. 6 , 23 (2020). Adela Alemu, M. et al. Antidiarrheal Effect of 80% Methanol Extract and Fractions of the Leaves of Ocimum lamiifolium in Swiss Albino Mice. Evidence-based Complement. Altern. Med. 2022 , (2022). Evans, W. C., Evans, D. & Trease, G. E. Trease and Evans Pharmacognosy . (Saunders/Elsevier, 2009). Additional Declarations Competing interest reported. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for profit sectors. But, the organization support the author by providing laboratory facilities, equipment's and chemical during laboratory analysis and Mr. Yohannes Shuka support the author in publication process. Individually, the author has no any need competing interests. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6429306","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":451952821,"identity":"e145af95-bbdc-44d7-90b2-b81ad387997a","order_by":0,"name":"Defar Bekele Adugna","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABBUlEQVRIie3PMUvDQBTA8Xc8yJQ2a+9bvFXwm7gkFNyuCl0y1HIgxCXuClK/QlyyeuEgWQ6yVhyM+AUqLhEUvegmpKmb4P2HNzzej+MAXK6/mGINAO2H4ANTENsNohwgSADHhx0BBaYjbBey0d+EJd1qgIyrU/YSE84CXyv1vDo5CM4saeO8l3CjkRvy5vw8CYvLvBIXmkmWmvteQusjxSX5UVb7pEd5KaQlyJIt5OEJXyVNots62Oj3q1JcD5I1evYVirJRCvZ4IbIhws3U25MUziempCItlbixpNj2l3Gl8U6+fcyCdPrYtIulWNW6aNq4n/xMf021871t+Ztjl8vl+id9AvJ7ZjyjJ1DqAAAAAElFTkSuQmCC","orcid":"","institution":"Madda Walabu University","correspondingAuthor":true,"prefix":"","firstName":"Defar","middleName":"Bekele","lastName":"Adugna","suffix":""}],"badges":[],"createdAt":"2025-04-11 14:38:31","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6429306/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6429306/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":82234495,"identity":"e2b69c0b-8f7c-44b5-931c-a4a3e46df111","added_by":"auto","created_at":"2025-05-08 06:47:47","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":196929,"visible":true,"origin":"","legend":"\u003cp\u003epartition by petroleum ether\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-6429306/v1/d3173a2ba9e4e62557d8e890.jpeg"},{"id":82234064,"identity":"0e837b27-2629-4d16-8b2a-b05c0c330a77","added_by":"auto","created_at":"2025-05-08 06:39:47","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":186444,"visible":true,"origin":"","legend":"\u003cp\u003epartition by ethylene acetate\u003c/p\u003e","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-6429306/v1/3b07ffbede6c38c249691380.jpeg"},{"id":82233126,"identity":"dc4594f6-395e-473a-a24f-73a58d3f7509","added_by":"auto","created_at":"2025-05-08 06:31:47","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":167022,"visible":true,"origin":"","legend":"\u003cp\u003epartition by n-butanol\u003c/p\u003e","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-6429306/v1/3ae8fa130a910ac6575cfd0d.jpeg"},{"id":84550536,"identity":"b9874edc-66ec-40bb-ae3f-c408661f0922","added_by":"auto","created_at":"2025-06-13 10:08:51","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1084046,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6429306/v1/18e06630-a6a7-4432-8da7-874ba0b6f817.pdf"}],"financialInterests":"Competing interest reported. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for profit sectors. But, the organization support the author by providing laboratory facilities, equipment's and chemical during laboratory analysis and Mr. Yohannes Shuka support the author in publication process. \nIndividually, the author has no any need competing interests.","formattedTitle":"Extraction and Phytochemical Screening of Secondary Metabolites from Leonotis Ocymifolia (Bokolu) plant on areal part","fulltext":[{"header":"Introduction","content":"\u003cp\u003eMedicinal plants are now more focused than ever because they have the capability of producing many undeniable benefits to people, especially in the line of medicine and\u003cbr\u003e pharmacological study. The medicinal power of these plants lies in phytochemical constituents that cause definite pharmacological actions on growth of pathogens \u003csup\u003e1\u003c/sup\u003e. Phytochemicals are natural compounds produced in plants such as alkaloids, glycosides, flavonoids, tannins, steroids, terpenoids and phenols are of secondary metabolites of plant that serves as defensives agents of microorganism\u0026rsquo;s \u003csup\u003e2\u003c/sup\u003e. Around worldwide, including Ethiopia traditional medicinal plant is more applicable to treat disease and infection starting from ancient time. The use of traditional medicine in Ethiopia is based on indigenous knowledge and has long been practiced in the treatment of various diseases \u003csup\u003e3\u003c/sup\u003e. For centuries, people have relied heavily on traditional medicine to treat a variety of physical and mental ailments. Approximately 80% of people in the country still rely on traditional medicine as their primary healthcare system \u003csup\u003e4\u003c/sup\u003e, and more than 95% consists of traditional herbal medicines. \u003c/p\u003e\n\u003cp\u003eIn Ethiopia, many plant species used as a medicinal plant and \u003cem\u003eLeonotis ocymifolia \u003c/em\u003eis typically used plant almost all part of population of Ethiopia \u003csup\u003e5\u003c/sup\u003e\u003cem\u003e. Leonotis ocymifolia \u003c/em\u003eis minaret flower (flowering plant), shrub and has along white corolla covered by orange rufous hair which is belong to Lamiaceae family \u003csup\u003e6\u003c/sup\u003e. In Ethiopia, it is known as Bokolu, and it is good traditional medicinal plant for many diseases. The plant is reasonably drought-resistant and wind tolerant. Unlike the similar \u003cem\u003eLeonotus leonuris,\u003c/em\u003e in the \u003cem\u003eocymifolia,\u003c/em\u003e the tubular flowers are bolder and larger \u003csup\u003e7\u003c/sup\u003e. \u003cem\u003eLeonotis ocymifolia\u003c/em\u003e is folk medicine which is used to treat mich and other belief in Ethiopia \u003csup\u003e2\u003c/sup\u003e\u003csup\u003e,\u003c/sup\u003e\u003csup\u003e8\u003c/sup\u003e. In society, the plant is used for the diarrheal, headache, cold (influenza), cattle doweling, stomach worm and malaria in some part of Ethiopia specially oromia region[Ethiomedicine]. The main objective of this study was to investigate in vitro isolation of secondary metabolites based on different polarity of solvent and phytochemical screening of secondary metabolites.\u003c/p\u003e"},{"header":"Results and Discussion","content":"\u003ch2\u003eExtraction yields\u003c/h2\u003e\n\u003cp\u003e6.0 g of grey crude was produced from the maceration of 100g of \u003cem\u003eL. ocymifolia\u003c/em\u003e plant powder. The process of partitioning the ethanol crude involved suspending (dissolving) the crude extracted in an aqueous solution, starting with the least polar solvent (petroleum ether), increasing the polarity of the solvents, and generating a separating layer due to the immiscibility of the two solvents. Following the partitioning of the ethanol crude into ethylene acetate, and n-buthanol crude, respectively, 2.15 g of yellowish ethylene acetate crude and 1.80 g of n-buthanol crude were ultimately produced. However, measuring petroleum ether is extremely difficult.\u0026nbsp;\u003c/p\u003e\n\u003ch2 id=\"_Toc159227245\"\u003ePartition\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eof Petroleum ether\u0026nbsp;\u003c/h2\u003e\n\u003cp\u003eEthanol crude extracts dissolve in aqueous solution and are partitioned within petroleum ether. Due to the immiscibility of the two solvents, a phase boundary is formed. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe upper layer (grey color) suspended over the aqueous solution (Figure 1) is petroleum ether crude solution and is carefully separated into a different container. The petroleum ether solution is concentrated by exposure at room temperature, and very few grams of crude were obtained.\u003c/p\u003e\n\u003ch2\u003ePartition of Ethylene acetate \u0026nbsp;\u003c/h2\u003e\n\u003cp\u003eEthylene acetate also partitioned within the aqueous ethanol solution, and phase boundary is formed in which the yellowish solution floated over aqueous. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe upper layer (yellowish) solution (Figure 2) is ethylene acetate, and the two layers separated. The ethylene crude solution is concentrated, and 2.15g of crude collected. In this partition, relatively high mass proportion of secondary metabolites revealed.\u003c/p\u003e\n\u003ch2\u003ePartition of n-buthanol\u003c/h2\u003e\n\u003cp\u003eFinally, the aqueous ethanol partitioned within n-butanol (Figure 3), and 1.80 g of crude collected. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe remaining aqueous solution discarded because of primary metabolites. Ethanol solvents can extracts from least polar compound to very more polar (primary metabolites). Therefore, partitioning is used to remove these primary metabolites from crude extract and narrow down the range of secondary metabolites based on their polarities.\u0026nbsp;\u003c/p\u003e\n\u003ch2 id=\"_Toc159227249\"\u003ePhytochemical Screening\u0026nbsp;\u003c/h2\u003e\n\u003cp\u003ePhytochemical screening of main secondary metabolites such as alkaloids, flavonoids, steroids, saponins, tannins, terpenoids, and phenols was studied on petroleum ether, ethylene acetate, and n-butanol crude partitions. In the ethylene acetate partition, terpenoids, phenols, flavonoids, and steroids were present, whereas phenols, glycosides, and tannins were present in the n-butanol partition (Table 1).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable1:\u003c/strong\u003e Table of Phytochemical Screening\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"635\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 111px;\"\u003e\n \u003cp\u003es/no\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 145px;\"\u003e\n \u003cp\u003ecomponents\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003epetroleum ether\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp\u003eethylene acetate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003en- buthanol\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 111px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 145px;\"\u003e\n \u003cp\u003ealkaloids\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 111px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 145px;\"\u003e\n \u003cp\u003eflavonoids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 111px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 145px;\"\u003e\n \u003cp\u003ephenols\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 111px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 145px;\"\u003e\n \u003cp\u003eglycosides\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 111px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 145px;\"\u003e\n \u003cp\u003eterpenoids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 111px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 145px;\"\u003e\n \u003cp\u003esaponins\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 111px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 145px;\"\u003e\n \u003cp\u003etannins\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 111px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 145px;\"\u003e\n \u003cp\u003esteroids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eIn general, the extraction of the aerial parts of \u003cem\u003eLeonotis ocymifolia\u003c/em\u003e in this study revealed secondary metabolites like alkaloids, flavonoids, phenols, glycosides, terpinoids, tannins, and steroids and the result similar within \u003csup\u003e3\u003c/sup\u003e. In contrast, saponins are not present in any of the crude oil partitions, and components are indicated by the \u0026quot;+\u0026quot; sign, while \u0026quot;-\u0026quot; denotes the absence of a component (Table 1). Table 1 show that the sugar component (primary metabolites) remained in the aqueous solution while nearly all of the secondary metabolites were detected in both solvents (ethyl acetate and n-butanol partition). The reason for this is that highly polar chemicals are attracted to water as a solvent.\u003c/p\u003e"},{"header":"Material and methodology","content":"\u003ch2\u003ePlant collection\u003c/h2\u003e\n\u003cp\u003eIn December 2024, the mature and healthy \u003cem\u003eLeonotis ocymifolia\u003c/em\u003e plants were gathered from a family garden in Goba woreda. Located in the Bale zone of the Oromia region, southeast of Addis Ababa, Ethiopia, lies the Goba woreda. The plant was authenticated by Botanist Reta Regassa, and a voucher specimen with the number BD00130/24 was deposited at the HU Herbarium, Hawassa University, for future reference. A sufficient number of plant sections will be collected, and the newly collected plants will undergo three rounds of washing in the laboratory, first with tap water to get rid of dust and then with distilled water to rinse. Next, after being cut with a knife into small pieces, it dries in the shade.\u0026nbsp;\u003c/p\u003e\n\u003ch2\u003eChemical and Material\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/h2\u003e\n\u003cp id=\"_Toc159227224\"\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eChemicals:\u0026nbsp;distilled water, ethanol (80%), ethyl acetate (AR), NaOH ,silica gel (mesh 200-400), petroleum ether(AR), naphthol, iodine, KI, HCl(37%), chloroform(AR), H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u0026nbsp;\u003c/sub\u003e(98%), FeCl\u003csub\u003e3\u003c/sub\u003e, methanol (80%).\u003c/p\u003e\n\u003cp\u003eMaterial:\u0026nbsp;Erlenmeyer flask, water bath shaker (SHZ -82), rotary evaporator (RE100-S), vacuum pump, column chromatography, TLC, dropper, spatula, electronic balance, Buchner funnel, watch man filter paper, iron stand up, electrical drier, Knife, separating funnel.\u003c/p\u003e\n\u003ch2\u003eExtraction\u0026nbsp;\u003c/h2\u003e\n\u003cp\u003eThe plant\u0026apos;s dried sections were ground using an electrical mill, and 100g of finely ground \u003cem\u003eLeonotis ocymifolia\u003c/em\u003e powder was soaked in 80% aqueous ethanol in 1:8 ratios for three days. The plant was then shaken for about 36 hours at 200\u0026deg;C using a water bath shaker (SHZ-82). Then watch man filter paper was used to separate the liquid. Grey ethanol crude was obtained by using a rotary evaporator (RE100-S) to concentrate the filtered liquid. Ultimately, ethanol crude was divided into petroleum ether and suspended in distilled water using a separating funnel. The two components were then carefully separated. Subsequently, the remaining aqueous solution was cautiously separated into separate containers after being gradually divided into ethylene acetate and n-buthanol. Ultimately, a rotary evaporator (RE100-S) was used to concentrate each partitioned solution independently.\u003c/p\u003e\n\u003ch2 id=\"_Toc159227232\"\u003ePhytochemicals screening\u0026nbsp;\u003c/h2\u003e\n\u003cp\u003ePetroleum ether crude, ethylene acetate, and n-butanol solutions were used to test the phytochemical components. Using standard procedure, this is done to determine whether secondary metabolites are present or absent\u0026nbsp;\u003csup\u003e9\u003c/sup\u003e\u003csup\u003e,\u003c/sup\u003e\u003csup\u003e10\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003ch2 id=\"_Toc159227233\"\u003eTest for alkaloids\u003c/h2\u003e\n\u003cp\u003eFew drops of Mayer\u0026rsquo;s reagent were added to 1ml of extract. A yellowish or white precipitate was formed, indicating the presence of alkaloids \u003csup\u003e10\u003c/sup\u003e.\u003c/p\u003e\n\u003ch2 id=\"_Toc159227234\"\u003eTest for flavonoids\u003c/h2\u003e\n\u003cp\u003eTwo to three drops of sodium hydroxide were added to 2 mL of extract. Initially, a deep yellow color appeared but it gradually became colorless by adding few drops of dilute HCL, indicating that flavonoids were present\u003csup\u003e10\u003c/sup\u003e.\u003c/p\u003e\n\u003ch2\u003e\u003cspan id=\"_Toc159227235\"\u003eTest for phenols\u003c/span\u003e\u0026nbsp;\u003c/h2\u003e\n\u003cp\u003e\u003cspan id=\"_Toc159227236\"\u003eTwo milliliters of 5% neutral ferric chloride solution were added to 1 ml of extract, the dark blue coloring indicating the presence of phenolic compounds \u003csup\u003e10\u003c/sup\u003e.\u003c/span\u003e\u003c/p\u003e\n\u003ch2\u003e\u003cspan id=\"_Toc159227237\"\u003eTest for glycosides\u003c/span\u003e\u003c/h2\u003e\n\u003cp\u003e\u003cspan id=\"_Toc159227238\"\u003eFew drops of Molisch\u0026rsquo;s reagent will be added to 2 ml of extract and will be shaken well. 2ml of conc. H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e will be added on the sides of the test tube. A reddish violet ring appeared at the junction of two layers immediately indicated \u003csup\u003e10\u003c/sup\u003e.\u003c/span\u003e\u003c/p\u003e\n\u003ch2\u003e\u003cspan id=\"_Toc159227239\"\u003eTest for terpenoids\u003c/span\u003e\u0026nbsp;\u003c/h2\u003e\n\u003cp\u003eAbout 5 mL of extract will be dissolved and mixed with 2 ml of chloroform, then 3 mL of concentrated H\u003csub\u003e2\u003c/sub\u003eSO4 were added and reddish brown coloration at the interface wills confirmed the presence of terpenoides\u0026nbsp;\u003csup\u003e10\u003c/sup\u003e.\u003c/p\u003e\n\u003ch2 id=\"_Toc159227240\"\u003eTest for tannins\u0026nbsp;\u003c/h2\u003e\n\u003cp\u003e0.1% FeCl\u003csub\u003e3\u003c/sub\u003e solution added to few ml of extract and the blue, blue-black, green or blue-green color which confirmed the presence of tannins\u0026nbsp;\u003csup\u003e10\u003c/sup\u003e.\u003c/p\u003e\n\u003ch2 id=\"_Toc156055473\"\u003eTest for sponins\u003c/h2\u003e\n\u003cp\u003eA drop of Na\u003csub\u003e2\u003c/sub\u003eCO\u003csub\u003e3\u003c/sub\u003e solution was added to 5 ml of extract in a test tube. After vigorous shaking, it was left to rest for five minutes. Foam formation indicated the presence of saponins\u0026nbsp;\u003csup\u003e10\u003c/sup\u003e.\u003c/p\u003e\n\u003ch2 id=\"_Toc159227241\"\u003eTest for steroids\u0026nbsp;\u003c/h2\u003e\n\u003cp\u003e2\u0026thinsp;ml of chloroform and concentrated H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e were added with the 5\u0026thinsp;ml aqueous plant crude extract. In the lower chloroform layer red color appeared that indicated the presence of steroids\u0026nbsp;\u003csup\u003e10\u003c/sup\u003e.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003e According to the results, the plant is rich within secondary metabolites and further experiments are required to comprehensively, characterized bioactive component and explore their antimicrobial activities.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eData availability\u003c/h2\u003e\n\u003cp\u003e\u0026nbsp;The data and material used to support the findings of this study are available main body text and are additional available from the corresponding author upon request.\u003c/p\u003e\n\u003ch2\u003eAcknowledgements\u0026nbsp;\u003c/h2\u003e\n\u003cp\u003eThe author is thankful to Madda Walabu University, Department of Chemistry, for providing laboratory facilities and chemical support during laboratory analysis. The author gratefully thanks Mr. Yohannes Shuka for supporting publication process. \u0026nbsp;\u003c/p\u003e\n\u003ch2\u003eAuthor contributions\u003c/h2\u003e\n\u003cp\u003e\u0026nbsp;Defar Bekele: conceptualization, writing (original draft), Conceived and designed the experiments methods; performed the experiments; Analyzed and interpreted the data; Contributed reagents, materials, analysis tools or data; Validation, Writing (review and editing), preparing figures, collecting data, validation investigation, and Formal analysis.\u0026nbsp;\u003c/p\u003e\n\u003ch2\u003eFunding\u003c/h2\u003e\n\u003cp\u003e\u0026nbsp;This research did not receive any specific grant from funding agencies in the public, commercial, or not-for profit sectors.\u003c/p\u003e\n\u003ch2\u003eCompeting interests\u0026nbsp;\u003c/h2\u003e\n\u003cp\u003eThe author declares no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eYadav, R. \u0026amp; Agarwala, M. Phytochemical analysis of some medicinal plants. \u003cem\u003eJ. Phytol.\u003c/em\u003e \u003cstrong\u003e3\u003c/strong\u003e, 10\u0026ndash;14 (2011).\u003c/li\u003e\n\u003cli\u003eAgidew, M. G. Phytochemical analysis of some selected traditional medicinal plants in Ethiopia. \u003cem\u003eBull. Natl. Res. Cent.\u003c/em\u003e \u003cstrong\u003e46\u003c/strong\u003e, (2022).\u003c/li\u003e\n\u003cli\u003eAyele, T. M., Abebe, E. C. \u0026amp; Kassie, A. B. Investigation of antibacterial and anti-diarrhoeal activities of 80% methanol leaf and fruit extract of leonotis ocymifolia (Burm. f) iwarsson (lamiaceae). \u003cem\u003eJournal of Experimental Pharmacology\u003c/em\u003e vol. 13 613\u0026ndash;626 at https://doi.org/10.2147/JEP.S319981 (2021).\u003c/li\u003e\n\u003cli\u003eDubale, S., Kebebe, D., Zeynudin, A., Abdissa, N. \u0026amp; Suleman, S. Phytochemical Screening and Antimicrobial Activity Evaluation of Selected Medicinal Plants in Ethiopia. \u003cem\u003eJ. Exp. Pharmacol.\u003c/em\u003e \u003cstrong\u003e15\u003c/strong\u003e, 51\u0026ndash;62 (2023).\u003c/li\u003e\n\u003cli\u003eAsteraye, Y. \u003cem\u003eADDIS ABABA UNIVERSITY OFFICE OF RESEARCH AND GRADUATE PROGRAMS PHYTOCHEMICAL INVESTIGATION ON THE LEAVES OF Leonotis Ocymifolia\u003c/em\u003e. (2006).\u003c/li\u003e\n\u003cli\u003eTeklu, T., Engidawork, E., Nedi, T., Teklehaymanot, T. \u0026amp; Gebremeskel, L. Evaluation of the Antimalarial Activity of the Hydroalcoholic Extract of Leaf of Leonotis ocymifolia (Burm. f.) Iwarsson (Lamiaceae) against Plasmodium berghei in Mice. \u003cem\u003eEvidence-based Complement. Altern. Med.\u003c/em\u003e \u003cstrong\u003e2020\u003c/strong\u003e, (2020).\u003c/li\u003e\n\u003cli\u003eA Review on Medicinal Plants Against Some Human Pathogenic Bacteria (E. coli, S.dysentery, S.typhi, P.aeruginosa and S. aureus) in South East Ethiopia. \u003cem\u003eJ. Biol. Agric. Healthc.\u003c/em\u003e (2019) doi:10.7176/jbah/9-19-03.\u003c/li\u003e\n\u003cli\u003eTalema, A. Review of Phytochemical Analysis of Selected Traditional Medicinal Plants in Ethiopia. \u003cem\u003eInt. J. Homeopath. Nat. Med.\u003c/em\u003e \u003cstrong\u003e6\u003c/strong\u003e, 23 (2020).\u003c/li\u003e\n\u003cli\u003eAdela Alemu, M. \u003cem\u003eet al.\u003c/em\u003e Antidiarrheal Effect of 80% Methanol Extract and Fractions of the Leaves of Ocimum lamiifolium in Swiss Albino Mice. \u003cem\u003eEvidence-based Complement. Altern. Med.\u003c/em\u003e \u003cstrong\u003e2022\u003c/strong\u003e, (2022).\u003c/li\u003e\n\u003cli\u003eEvans, W. C., Evans, D. \u0026amp; Trease, G. E. \u003cem\u003eTrease and Evans Pharmacognosy\u003c/em\u003e. (Saunders/Elsevier, 2009).\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":"phytochemical, extraction, partition, isolation","lastPublishedDoi":"10.21203/rs.3.rs-6429306/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6429306/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eNowadays, the uses of medicinal plants are more diverse to prevent human health care, in which medicinal plants are powerful inhibition of microorganisms cause disease. The study is conducted to assess extraction, phytochemical screening and methods of extracting secondary metabolites of a traditionally used medicinal plant, Leonotis ocymifolia. The plant has been extracted using 80% aqueous ethanol by maceration technique, and 6.0 g of grey ethanol crude were obtained. The crude ethanol suspended in water and partitioned within different polarity of solvents (petroleum ether, ethylene acetate, and n-buthanol), respectively. During partitioning, 2.15 g of ethylene acetate were collected after concentrated using a rotary evaporator and few grams of petroleum ether were obtained. The study revealed phytochemical components like alkaloids, flavonoids, phenols, tannins, glycosides, terpeniods and steroids.\u003c/p\u003e","manuscriptTitle":"Extraction and Phytochemical Screening of Secondary Metabolites from Leonotis Ocymifolia (Bokolu) plant on areal part","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-08 06:31:42","doi":"10.21203/rs.3.rs-6429306/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":"bec4e73c-c2a8-4fe9-8299-0ef7ade88f19","owner":[],"postedDate":"May 8th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":48066134,"name":"Physical sciences/Chemistry"},{"id":48066135,"name":"Physical sciences/Materials science"},{"id":48066136,"name":"Physical sciences/Nanoscience and technology"}],"tags":[],"updatedAt":"2025-06-13T10:08:31+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-08 06:31:42","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6429306","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6429306","identity":"rs-6429306","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}