Extraction and chemical characterization of neem seed oil (Azadirachta indica)
DOI:
https://doi.org/10.20873/jbb.uft.cemaf.v11n4.16258Keywords:
Azadirachta indica a. juss , microextraction in solid phase headspace, gas chromatography, sohxletAbstract
The neem tree (Azadirachta indica A. Juss) is known for its therapeutic, pesticide, fertilizer and pharmacological properties. Due to these attributes, this tree has economic potential and continues to be used worldwide. For oil extraction, the Sohxlet system was used, for the extraction and characterization of volatiles, the HS-SPME/GC-MS method was used, and the fatty acid profile was characterized by GC-FID. The average yield obtained from the oil was 23%. A total of 34 volatile compounds were detected, of which 9 were identified by Kovat's index: 2-methyldecane, 3-methyldecane, 4-methyldecane, 2-methylundecane, 3-methylundecane, 3,4-dimethylthiophene, 2,5-diol- 3-hexane, 1,2,3-trimethylbenzene and buticyclohexane. In addition to the volatiles in the oil, 7 fatty acids were found; oleic acid, linoleic acid, palmitic acid and stearic acid, with the oil extracted from the seeds having the highest concentration of oleic acid and linoleic acid in commercial neem oil. Given the presence of these chemical compounds in neem oil, it is suggested, for subsequent investigations, the isolation and application of organosulfur compounds as insecticides. The extraction of oil from flowers and neem seeds with solvents that are less harmful to the environment, together with its application, makes it possible to evaluate its pesticide potential. In addition, it is essential to evaluate the influence of neem on the bee population, analyzing cases of mortality and seeking solutions to avoid environmental damage. Additionally, the viability of neem oil as an industrial and hospital cleaning product is highlighted, exploring antimicrobial, antibacterial and biodegradable properties.
References
Alba SL, García-Gonzáles C, Coronado OMA, Ayla BJR, Alpírez GM, Montes DGLNN. extraction methods and ap-plications of bioactive compounds from neem (Azadirachta indica): A mini-review. Mini-Reviews in Organic Chemis-try, v.20, n.7, p.644-654, 2023. https://doi.org/10.2174/1570193X19666220707125726
Aider FA, Kellouche A, Fellag H, Debras JF. Evaluation of the bio-insecticidal effects of the main fatty acids of olive oil on Callosobruchus maculatus F. (Coleoptera-Bruchidae) in cowpea (Vigna unguiculata (L.). Journal of Plant Diseases and Protection, v.123, p.235-245, 2016. https://doi.org/10.1007/s41348-016-0034-z
Ayalew AA. Chromatographic and spectroscopic determina-tion of solvent-extracted Lantana camara leaf oil. Journal of International Medical Research, v.48, n.10, p.1-12, 2020.
https://doi.org/10.1177/0300060520962344
Ayinde AA, Morakinyo OM, Sridhar MKC. Repellency and larvicidal activities of Azadirachta indica seed oil on Anop-heles gambiae in Nigeria. Heliyon, v.6, n.5, p. 1-7, 2020.
https://doi.org/10.1016/j.heliyon.2020.e03920
Baby AR, Freire TB, Marques GA, Rijo P, Lima FV, Carva-lho JCM, Rojas P, Magalhães WV, Velasco MVR, Moro-cho-Jácome AL. Azadirachta indica (Neem) as a potential natural active for dermocosmetic and topical products: a nar-rative review. Cosmetics, v. 9, n. 3, p.1-17, 2022. https://doi.org/10.3390 /cosmetics9030058
Balandrin MF, Lee SM, Klocke JA. Biologically active volatile organosulfur compounds from seeds of the neem tree, Aza-dirachta indica (Meliaceae). Journal of Agricultural and Fo-od Chemistry, v.36, n.5, p.1048–1054, 1988. https://doi.org/10.1021/jf00083a035
Barbosa LMP, Santos JO, Sousa RCM, Furtado JLB, Vidinha P, Garcia MAS, Vitorino HA, Dall’Oglio DF. Bioherbicide from Azadirachta indica seed waste: exploitation, efficient extraction of neem oil and allelopathic effect on senna occi-dentalis. Recycling, v. 8, n. 3, p.1-15, 2023. https://doi.org/10.3390/recycling 8030050
Beyecha Hundi̇e K, Abdi̇ssa D, Bekele Bayu A. Extraction, optimization, and characterization of neem seed oil via box-behnken design approach. Journal of the Turkish Chemical Society Section A: Chemistry, v.9, n.2, p.513-526, 2022.
https:// doi.org/10.18596/jotcsa.1039997
Breu W. Allium cepa L. (Onion) Part 1: Chemistry and analysis. Phytomedicine, v.3, n.3, p.293–306, 1996. https://doi.org/10.1016/S0944-7113(96)80069-9
Bukvicki DR, Tyagi AK, Gottardi DG, Veljic MM, Jankovic SM, Guerzoni ME et al. Assessment of the chemical com-position and in vitro antimicrobial potential of extracts of the liverwort scapania aspera. Natural Product Communications, v.8, n.9, p.1-4, 2013. https://doi.org/10.1177/1934578X1300800932
Cantrell MS, Seale JT, Arispe SA, McDougal OM. Determi-nation of Organosulfides from Onion Oil. Foods, v.9, n.7, p.884, 2020.
https://doi.org/10.3390/foods9070884
Carvalho AMS, Heimfarth L, Pereira EWM, Oliveira FS, Menezes IRA, Coutinho HDM et al. Phytol, a chlorophyll component, produces antihyperalgesic, anti-inflammatory, and antiarthritic effects: possible NFκB pathway involve-ment and reduced levels of the proinflammatory Cytokines TNF-α and IL-6. Journal of Natural Products, v.83, n.4, p.1107-1117, 2020. https://doi.org/10.1021/acs.jnatprod.9b01116
Casillas-vargas G, Ocasio-Malavé C, Medina S, Morales-Guzman C, Valle RG Del, Carballeira NM, Carballeira NM, Sanabria-Ríos DJ. Antibacterial fatty acids: An update of possible mechanisms of action and implications in the deve-lopment of the next-generation of antibacterial agents. Pro-gress in Lipid Research, v.82, p.1-10, 2021. https://doi.org/10.1016/j.plipres.2021.101093
Chaudhary MF, Ashraf A, Waseem M, Hayat S, Nadeem HU, Siddique MH et al. Neem oil. In: Inamuddin, Boddula R, Asiri AM (org.). Green Sustainable Process Chemical and Environmental. Elsevier, 2021. p.57-73.
Chinnasamy G, Chandrasekharan S, Koh TW, Bhatnagar S. Synthesis, characterization, antibacterial and wound healing efficacy of silver nanoparticles from Azadirachta indica. Frontiers in Microbiology, v.12, p.1-14, 2021. https://doi.org/10.3389/fmicb.2021.611560
Choi SM, Lee D-J, Kim J-Y, Lim S-T. Volatile composition and sensory characteristics of onion powders prepared by convective drying. Food Chemistry, v.231, p.386-392, 2017.
https://doi.org/10.1016/j.foodchem.2017.03.129
Desai SN, Jadhav AJ, Holkar CR, Pawar BG, Pinjari DV. Extraction and microencapsulation of Buchanania lanzan Spreng seed oil. Chemical Papers, v.76, n.6, p.3521-3530, 2022.
https://doi.org/10.1007/s11696-022-02116-0
Ebrahimi M, Safaralizade MH, Valizadegan O. Contact toxi-city of Azadirachta indica (Adr. Juss.), Eucalyptus camal-dulensis (Dehn.) and Laurus nobilis (L.) essential oils on mortality cotton aphids, Aphis gossypii Glover (Hem.: Aphididae). Archives of Phytopathology and Plant Protec-tion, v.46, n.18, p.2153–2162, 2013. https://doi.org/10.1080/03235408.2013 .774526
Fagbemi KO, Aina DA, Olajuyigbe OO. Soxhlet extraction versus hydrodistillation using the clevenger apparatus: A comparative study on the extraction of a volatile compound from Tamarindus indica seeds. The Scientific World Jour-nal, v.2021, p.1-8, 2021. https://doi.org/10.1155/2021/5961586
Fernandes SR, Barreiros L, Oliveira RF, Cruz A, Prudêncio C, Oliveira AI, Pinho C, Santos N, Morgado J. Chemistry, bioactivities, extraction and analysis of azadirachtin: State-of-the-art. Fitoterapia, v.134, p.141-150, 2019. https://doi.org/10.1016/j.fitote.2019.02.006
Ghosh S, Mali SN, Bhowmick DN, Pratap AP. Neem oil as natural pesticide: Pseudo ternary diagram and computational study. Journal of the Indian Chemical Society, v.98, n.7, p.1-7, 2021.
https://doi.org/10.1016/j.jics.2021.100088
Granado-Casas M, Mauricio D. Oleic acid in the diet and what it does: implications for diabetes and its complications. In: Watson RR, Preedy VR (ed.). Bioactive food as dietary in-terventions for diabetes. Academic Press. 2019. p.211-229.
https://doi.org/ 10.1016/ B978-0-12-813822-9.00014-X
Guchhait KC, Manna T, Barai M, Karmakar M, Nandi SK, Jana D, Dey A, Panda S, Raul P, Patra A, Bhattacharya R, Chatterjee S, Panda Ak, Ghosh C. Antibiofilm and antican-cer activities of unripe and ripe Azadirachta indica (neem) seed extracts. BMC Complementary Medicine and Therapi-es, v.22, n.1, p.1-18, 2022. https://doi.org/10.1186/s12906-022-03513-4
Gutbrod K, Romer J, Dörmann P. Phytol metabolism in plants. Progress in Lipid Research, v.74, p.1-17, 2019. https://doi.org/10.1016/j.plipres.2019.01.002
Hirondart M, Rombaut N, Fabiano-Tixier AS, Bily A, Chemat F. Comparison between pressurized liquid extraction and conventional soxhlet extraction for rosemary antioxidants, yield, Composition, and Environmental Footprint. Foods, v.9, n.5, p.1-14, 2020.
https://doi.org/10.3390/foods9050584
Hossain MA, Al-Toubi WAS, Weli AM, Al-Riyami QA, Al-Sabahi JN. Identification and characterization of chemical compounds in different crude extracts from leaves of Omani neem. JTUSCI, v.7, n.4, p.181-188, 2013. https://doi.org/10.1016/j.jtusci .2013.05.003
Islam MD T, Alencar MVOB de, Machado KC, Machado KC, Melo-Cavalcante A A de C, Sousa DP de et al. Phytol in a pharma-medico-stance. Chemico-Biological Interactions, v.240, p.60–73, 2015. https://doi.org/10.1016/j.cbi.2015.07.010
Islam MT, Ali ES, Uddin SJ, Shaw S, Islam Md A, Ahmed Md I et al. Phytol: A review of biomedical activities. Food and Chemical Toxicology, v.121, p.82-94, 2018. https://doi.org/ 10.1016/j.fct.2018.08.032
Islas JF, Acosta E, G-Buentello Z, Delgado-Gallegos JL, Moreno-Treviño MG, Escalante B et al. An overview of neem (Azadirachta indica) and its potential impact on health. Journal of Functional Foods, v.74, p.1-13, 2020. https://doi.org/10.1016/j.jff.2020.104171
Ismaila SS, Sani Y, Sani AA, Yakasai SM, Momoh H, Mohammed Sa’a. E. Determination of fatty acids and physi-cochemical properties of neem (Azadrachta Indica L) seed oil extracts. AJOL, v.8, n.1, p.149-160, 2022. https://doi.org/10.4314/dujo pas.v8i1a.16
Jeelani I et al. Pharmaceutical potential of constituents from Azadirachta indica and their specific role as anti-cancer agents. Current Bioactive Compounds, v.19, n.3, p.94-103, 2023. http://dx.doi.org/10.2174/1573407218666220818110059
Jishma P, Hussain N, Chellappan R, Rajendran R, Mathew J, Radhakrishnan EK. Strain-specific variation in plant growth promoting volatile organic compounds production by five different Pseudomonas spp. as confirmed by response of Vigna radiata seedlings. Journal of Applied Microbiology, v.123, n.1, p.204–216, 2017. https://doi.org/10.1111/jam.13474
Kaura SK, Gupta SK, Chowdhury JB. Morphological and oil content variation in seeds of Azadirachta indicates A. Juss. (Neem) from northern and western provenances of India. Plant Foods for Human Nutrition, v.52, p.293-298, 1998. https://doi.org/10.1023/A: 1008013424150
Keneni YG, Marchetti JM. Oil extraction from plant seeds for biodiesel production. AIMS Energy. v. 5, n. 2, p. 316-340, 2017.
https://doi.org/10.3934/energy.2017.2.316
Kilani-Morakchi S, Morakchi-Goudjil H, Sifi K. Azadirachtin-Based Insecticide: Overview, Risk Assessments, and Future Directions. Frontiers in Agronomy, v.3, p.1-13, 2021.
https://doi.org/10.3389/fagro.2021.676208
Kostik V, Memeti S, Bauer B. Fatty acid composition of edible oils and fats. Journal of Hygienic Engineering and Design. v.4, p.112-116, 2013.
Koul O. Biological activity of volatile di-n-propyl disulfide from seeds of neem, Azadirachta indica (Meliaceae), to two species of stored grain pests, Sitophilus oryzae (L.) and Tri-bolium castaneum (Herbst). Journal of Economic Entomo-logy, v.97, n.3, p.1142-1147, 2004. https://doi.org/10.1093/jee/97.3.1142
Kumar S, Singh N, Devi LS, Kumar S, Kamle M, Kumar P et al. Neem oil and its nanoemulsion in sustainable food pre-servation and packaging: Current status and future pros-pects. Journal of Agriculture and Food Research, v.7, p.1-9.
https://doi.org/10.1016/j.jafr.2021.100254
Kumar Tyagi A, Bukvicki D, Gottardi D, Veljic M, Guerzoni ME et al. Antimicrobial potential and chemical characteriza-tion of serbian liverwort (Porella arboris-vitae): SEM and TEM Observations. Evidence-Based Complementary and Alternative Medicine, v.2013, p.1-7, 2013. https://doi.org/10.1155/2013/382927
Lienard V, Seck D, Lognay G, Gaspar C, Severin M. Biologi-cal activity of Cassia occidentalis L. against Callosobruchus maculatus (F.) (Coleoptera: Bruchidae). Journal of Stored Products Research, v.29, n.4, p.311-318, 1993. https://doi.org/10.1016/0022-474X(93)90046-7
Loganathan T, Barathinivas A, Soorya C, Balamurugan S, Nagajothi TG, Ramya S, et al. Physicochemical, druggable, ADMET pharmacoinformatics and therapeutic potentials of azadirachtin - a prenol lipid (triterpenoid) from seed oil ex-tracts of Azadirachta indica A. Juss. Journal of Drug Deli-very and Therapeutics, v.11, n.5, p.33-46, 2021. https://doi.org/10.22270/jddt.v1 1i5.4981
López-Bascón MA, Luque de Castro MD. Soxhlet Extraction. In: Poole CF (org.). Liquid-Phase Extraction. Elsevier. 2020. p.27 https://doi.org/10.10 16/B978-0-12-816911-7.00011-6
Marangoni F, Agostoni C, Borghi C, Catapano AL, Cena H, Ghiselli A, Vecchia CL, Lercker G, Manzato E, Pirillo A, Riccardi G, Risé P, Visioli F, Poli A. Dietary linoleic acid and human health: Focus on cardiovascular and cardiometa-bolic effects. Atherosclerosis, v.292, p.90–98, 2020. https://doi.org/10.1016/j.atherosclerosis.2019 .11.018
Mohamadpoor M, Amini J, Ashengroph M, Azizi A. Evalua-tion of biocontrol potential of Achromobacter xylosoxidans strain CTA8689 against common bean root rot. Physiologi-cal and Molecular Plant Pathology, v.117, p.101769, 2022.
https://doi.org/10.1016/j.pmpp.2021.101769
Mubarak AM, Kulatilleke CP. Sulphur constituents of neem seed volatiles: A revision. Phytochemistry, v.29, n.10, p.3351-3352, 1990. https://doi.org/10.1016/0031-9422(90)80213-Z
Mulla M, Ahmed J, Alagarsamy S, K Habeebullah SF. Utili-zation of novel and rapid techniques for characterization of neem Azadirachta indica seed oil and palm oil blends. Inter-national Journal of Food Engineering, v.16, n.10, p.20200047, 2020.
https://doi.org/10.1515/ijfe-2020-0047
Neves EJM, Carpanezzi AA. The cultivation of neem (Aza-dirachta indica) for fruit production in Brazil. Colombo: Embrapa Forests, 8p. 2008.
Olofsson P, Hultqvist M, Hellgren LI, Holmdahl R. Phytol: A chlorophyll component with anti-inflammatory and metabo-lic properties. In: Jacob C, Kirsch G, Slusarenko A, Win-yard P, Burkholz T, (org.). Recent advances in redox active plant and microbial products: From basic chemistry to widespread applications in medicine and agriculture. Dor-drecht: Springer Netherlands. 2014. p.14.
Perera AGWU, Karunaratne MMSC, Chinthaka SDM. Bioac-tivity and volatile profiling of Azadirachta indica leaves for the management of maize weevil, Sitophilus zeamais (motsch.) infestations. JTFE, v.8, n.1, p.10-24, 2018. https://doi.org/10.31357/jtfe.v8i1 .3479
Perera WPRT, Liyanage JA, Dissanayake KGC, Gunathilaka H, Weerakoon WMTDN, Wanigasekara DN et al. Antiviral potential of selected medicinal herbs and their isolated natu-ral products. BioMed Research International, v.2021, p.1-18, 2021. https://doi.org/10.1155/2021/7872406
Ragunath C, Kumar YAS, Kanivalan I, Radhakrishnan S. Phytochemical screening and GC-MS analysis of bioactive constituents in the methanolic extract of Caulerpa racemosa (Forssk.) J. Agardh and Padina boergesenii Allender & Kraft. Current Applied Science And Technology, v.20, n.3, p.380-393, 2020.
Rana S, Saxena A. Medicinal chemistry of neem: A state of art. Kochi, Kerala, India: 1, 2023. https://doi.org/10.1063/5.0114567
Raza W, Ling N, Liu D, Wei Z, Huang Q, Shen Q. Volatile organic compounds produced by Pseudomonas fluorescens WR-1 restrict the growth and virulence traits of Ralstonia solanacearum. Microbiological Research, v.192, p.103-113, 2016.
https://doi.org/10.1016/j.mi cres.2016.05.014
Saha tchinda J-B, Tchebe TMF, Tchoukoua A, Yona AMC, Fauconnier ML, Kor M N, et al. Fatty acid profiles, antioxi-dant, and phenolic contents of oils extracted from Acacia polyacantha and Azadirachta indica (Neem) seeds using green solvents. Journal of Food Processing and Preserva-tion, v.45, n.2, p.1-13, 2021. https://doi.org/10.1111/jfpp.15115
Sarkar S, Singh RP, Bhattacharya G. Exploring the role of Azadirachta indica (neem) and its active compounds in the regulation of biological pathways: an update on molecular approach. 3 Biotech, v.11, n.4, p.178, 2021. https://doi.org/10.1007/s13205-021-02745-4
Satyal P, Craft JD, Dosoky NS, Setzer WN. The chemical compositions of the volatile oils of garlic (Allium sativum) and wild garlic (Allium vineale). Foods, v.6, n.8, p.63, 2017.
https://doi.org/10.3390/foods6080063
Shariare MH, Noor HB, Khan JH, Uddin J, Ahamad SR, Altamimi MA, Alanazi FK, Kazi M. Liposomal drug deli-very of Corchorus olitorius leaf extract containing phytol using design of experiment (DoE): In-vitro anticancer and in-vivo anti-inflammatory studies. Colloids and Surfaces B: Biointerfaces, v.199, p,1-11, 2021. https://doi.org/10.1016/j.colsurfb.2020.111543
Suganthy M, Gajendra CV. Chemical characterization of Strychnos nux-vomica L. leaves for biopesticide properties using GC-MS. International Journal of Chemical Studies, v.8, n.1, p.1112-1116, 2020. https://doi.org/10.22271/chemi.2020.v8.i1o.8398
Tesfaye B, Tefera T, Misikir O, Tsegaye G. Extraction and comparison of essential oil from neem seed by using soxhlet extraction and simple distillation methods. International Journal of Engineering Technologies and Management Re-search, v.5, n.9, p.74-81, 2018. https://doi.org/10.29121/ijetmr.v5.i9.2018 .291
Tesfaye B, Tefera T. Extraction of essential oil from neem seed by using soxhlet extraction methods. IJAEMS, v.3, n.6, p.646-650, 2017.
https://dx.doi.org/10.24001/ij aems.3.6.5
Tulashie SK, Adjei F, Abraham J, Addo E. Potential of neem extracts as natural insecticide against fall armyworm (Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctui-dae). Case Studies in Chemical and Environmental Engi-neering, v.4, p.1-7, 2021.
https://doi.org/10.1016 /j.cscee.2021.100130
Ungo-Kore HY, Ibrahim YKE, Tytler BA. Anti-dermatophytic activity of hexane extracts of Azadirachta indicates A. Juss. African Journal of Microbiology Research, v.13, n.26, p.421-429, 2019. https://doi.org/10.5897/AJMR2018.8819
Winnett V, Sirdaarta J, White A, Clarke FM, Cock IE. Inhibi-tion of Klebsiella pneumoniae growth by selected australian plants: natural approaches for the prevention and manage-ment of ankylosing spondylitis. Inflammopharmacology, v.25, p.223-235, 2017. https://doi.org/10.1007/s10787-017-0328-1
Yami AM, Ibrahim ME, Raji A. Extraction and characterizati-on of oil from neem and yellow oleander seeds for biodiesel production. European Journal of Materials Science and En-gineering, v.5, n.4, p.212-221, 2020. https://doi.org/10.36868/ejmse.2020.05.04.212
Zambiazi RC, Przybylski R, Zambiazi MW, Mendonça CB. Fatty acid composition of vegetable oils and fats. Bulletin of the Center for Food Research and Processing, v.25, n.1, p.111-120, 2007.
http://dx.doi.org/10.5380/cep.v25i1.8399
Zatelli A, Fondati A, Maroli M. The knowns and unknowns of the efficacy of neem oil (Azadirachta indica) used as a preventative measure against Leishmania sand fly vectors (Phlebotomus genus). Preventive Veterinary Medicine, v.202, p.1-5, 2022.
https://doi.org/10.1016/j.prevetmed.2022.105618
Zhang H, Mallik A, Zeng RS. Control of panama disease of banana by rotating and intercropping with chinese chive (Al-lium Tuberosum Rottler): role of plant volatiles. Journal of Chemical Ecology, v.39, n.2, p.243-252, 2013. https://doi.org/10.1007/s10886-013-0243-x
Zhao B, Li H, Lan T, Wu D, Chen Z. Characterization of the chemical composition of chinese Moringa oleifera seed oil. Journal of the American Oil Chemists' Society, v.96, n.5, p.523-533, 2019.
https://doi.org/10.1002/aocs.12203
Zhao Y, Smyth HE, Tao K, Henry RJ, Gilbert RG. Starch molecular structural features and volatile compounds affec-ting the sensory properties of polished australian wild rice. Foods, v.11, n.4, p.1-22, 2022.
https://doi.org/10.3390/foods11040511
Zheng L, Shen X, Wang Y, Liang J, Xu M, Fang X, Gang C, Guilliang C. Content and composition of phospholipids, fatty acids and sterols in commercial natural phospholipid excipients. Current Pharmaceutical Analysis, v.17, n.7, p.892-902, 2021.
https://doi.org/10.2174 /1573412916999200605162707
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2023 Simyão Macêdo Cabral, Flamys Lena do Nascimento Silva, Pablo José Rodrigues, Magale Karine Diel Rambo, Jéssyka Ribeiro Santos, Mariana Helena Chaves, Bernardo Ferreira Pinto
This work is licensed under a Creative Commons Attribution 4.0 International License.
Copyright (c) 2024 - Journal of Biotechnology and Biodiversity
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License (CC BY 4.0 at http://creativecommons.org/licenses/by/4.0/) that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
Authors are permitted and encouraged to post their work online (e.g. in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (Available at The Effect of Open Access, at http://opcit.eprints.org/oacitation-biblio.html).