A study of the composition and antimicrobial activities of stored and freshly extracted leaf essential oil of Ocimum gratissimum L.

Document Type: Original Article


Department of Pharmaceutics, Obafemi Awolowo University, Ile-Ife, Nigeria


There have been concerns expressed on the biological activities of certain essential oils following long term storage. The aim of this study was therefore to determine the composition and antimicrobial activities of samples of stored and freshly extracted essential oil of Ocimum gratissimum Linneo (Lamiaceae) in order to provide comparison of these properties in the oils. The essential oils were obtained from steam distillation of fresh leaves of O. gratissimum collected in Ile-Ife (Nigeria) in the early morning hours in April 2005 and March 2013. These essential oils were then stored in amber coloured bottle in a refrigerator at 4 oC. GC-MS analyses of the stored and fresh essential oils identified twelve and five constituents respectively. The proportions in the stored and fresh essential oils were: thymol (46.2 and 76.0%, respectively), linalool (14.0 and 7.0%), neral (8.4 and 6.7%), eucalyptol (3.9 and 5.3%), terpinen-4-ol (1.8 and 2.4%), geranial (11.7 and 0%), α-terpineol (2.1 and 0%) and nerol (1.8 and 0%). The antibacterial activities of the two essential oils were comparable, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of which were from 0.25%v/v to 4.0%v/v. On the other hand, the stored essential oil showed higher antifungal effects with MIC and minimum fungicidal concentration (MFC) of <0.0625%v/v against Candida spp. Both essential oils completely killed 1.0 x 108cfu/ml bacterial innoculum within three hours. The differences in the antimicrobial activities obtained for the fresh and stored essential oil of O. gratissimum were presumably due to different proportion of constituents.

Graphical Abstract

A study of the composition and antimicrobial activities of stored and freshly extracted leaf essential oil of Ocimum gratissimum L.


Main Subjects

Adams, R.P., 2017. Identification of Essential Oil Components by Gas Chromatography/Mass Spectroscopy, Ed. 4.1. http://essentialoilcomponentsbygcms.com.

Akinkunmi, E.O., Oladele, A., Esho, O., Odusegun, I., 2015. Effects of storage time on the antimicrobial activities and composition of lemon grass oil. J. Appl. Res.  Med. Arom. Plants. 3, 105-111.

Aiyelabola, T., Ojo, I., Akinkunmi, O., 2012. Structural and antimicrobial studies of coordination compounds of phenylalanine and glycine. Int. J. Chem. 4, 49-59.

Ajileye, O.O., Obuotor, E.M., Akinkunmi, E.O., Aderogba, M.A., 2015. Isolation and characterization of antioxidant and antimicrobial compounds from Anacardium occidentale L. (Anacardiaceae) leaf extract. J. King Saud Univ. 12, 004.

Alarcón, L., Peña, A., Velasco, J., Baptista, J.G., Rojas, L., Aparicio, R., Usubillaga, A., 2015. Chemical composition and antibacterial activity of the essential oil of Ruilopezia bracteosa. Nat. Prod. Com. 10(4), 655-656.

Ali, M.E., Shamsuzzaman, L.A.M., 1968. Investigations on Ocimum gratissimum Linn. III, Constituents of the essential oil. Sci. Res. (Dacca). 5, 91-94.

Börje, A., Sköld, M., Matura, M., Karlberg, A.T., 2004. Autoxidation of the fragrance caryophyllene forms contact allergens. Contact Derm. 50(3), 191-192.

Bråred-Christensson, J., Forsström, P., Wennberg, A.M., Karlberg, A.T., Matura, M., 2009. Air oxidation increases skin irritation from fragrance terpenes. Contact Derm. 60(1), 32-40.

British Pharmacopoeia, 1988. London, Her Majesty’s Stationary Office, A137- A138.

Burfield, T., 2004. Notes on oxidation of oils in aromatherapy. Aromather. Today 1(62), 9-10.

Camilo, C.J., AlvesNonato, C.d.F., Galvao-Rodrigues, F.F., Costa, W.D., Clemente, G.G., Sobreira Macedo, M.A.C., Galvao Rodrigues, F.F., da Costa, J.G.M., 2017. Acaricidal activity of essential oils: a review. Trends Phytochem. Res. 1(4), 183-198.

Charles, D.J., Simon, J.E., 1992. A new geraniol chemotype of O. gratissimum L. J. Essent. Oil Res. 4, 231-234.

Chen, Y-H., Chiu, Y-W., Shyu, J-C., Tsai, C-C., Lee, H-H., Hung, C-C., Hwang, J-M., Liu, J-Y., Wang, W-H., 2015. Protective effects of Ocimum gratissimum polyphenol extract on carbon tetrachloride-induced liver fibrosis in rats. Chin. J. Physiol. 58(1), 55-63.

Figueiredo, A.C., Barroso, J.G., Pedro, L.G., Scheffer, J.J.C., 2008. Factors affecting secondary metabolite production in plants: volatile components and essential oils. Flav. Frag. J. 23, 213-226.

Fun, C.E., Baerheim Svendsen, A., 1990. Composition of Ocimum basilicum var. canum Sims and O. gratissimum L grown on Aruba. Flav. Frag. J. 5, 173-177.

Guenther, E., 1948. The Essential Oils. VoI II. Van Nostrand Co., New York.

Hegnauer, R., 1967. Chemotaxonomieder Pflanzen. BirkhauserVerlag, Basel 4, 314.

Joshi, R.K., 2017. GC-MS analysis of the essential oil of Ocimum gratissimum L. growing desolately in South India. Acta Chromatogr. 29(1), 111-119.

Kalita, J., Khan, M.L., 2013. Commercial potentialities of essential oil of Ocimum members growing in North East India. Int. J. Pharm. Life Sci. 4(4), 2559-2567.

Lamikanra, A., 2010. Essential Microbiology. Third Edition, Amkra Books, Lagos, Nigeria.

Lawrence, B.M., 1992. Labiatae oils: Mother Nature's Chemical Factor. Essential oils 19881991, Edit., B.M. Lawrence, Allured Publ., Stream,, 1, 88-206.

Lexa, G.M., Josphat, C.M., Francis, N.W., Titus, K.M., 2007. Chemical composition and antimicrobial activity of the essential oil of Ocimum gratissimum L. growing in Eastern Kenya. Afr. J. Biotech. 6(6), 760-765.

Mohammadhosseini, M., 2017. The ethnobotanical, phytochemical and pharmacological properties and medicinal applications of essential oils and extracts of different Ziziphora species. Ind. Crops. Prod. 105, 164-192.

Mohammadhosseini, M., Akbarzadeh, A., Hashemi-Moghaddam, H., Mohammadi Nafchi, A., Mashayekhi, H.A., Aryanpour, A., 2016. Chemical composition of the essential oils from the aerial parts of Artemisia sieberi by using conventional hydrodistillation and microwave assisted hydrodistillation: A comparative study. J. Essent. Oil. Bearing Plants. 19(1), 32-45.

Mohammadhosseini, M., Sarker, S.D., Akbarzadeh, A., 2017. Chemical composition of the essential oils and extracts of Achillea species and their biological activities: A review. J. Ethnopharmacol. 199, 257-315.

Oladimeji, F.A., Orafidiya, L.O., 2007. A study on the autoxidation profile of leaf essential oil of Lippia multiflora Moldenke under different storage conditions. Int. J. Essent. Oil Ther. 1, 39-44.

Olugbade, T.A., Kolipha-Kamara, M.I., Elusiyan, C.A., Onawunmi, G.O., Ogundaini, A.O., 2017. Essential oil chemotypes of three Ocimum species found in Sierra Leone and Nigeria. Med. Aromat. Plants. 6, 284, doi: 10.4172/2167-0412.1000284.

Onawunmi, G.O., 1989. Evaluation of the antimicrobial activities of citral. Lett. Appl. Microbiol. 9, 105-108.

Orafidiya, L.O., Adesina, S.K. Jr, Igbeneghu, O.A., Akinkunmi, E.O., Adetogun, G.E., Salau, A.O., 2006. The effect of honey and surfactant type on the antibacterial properties of the leaf essential oil of Ocimum gratissimum Linn. against common wound infecting organisms. Int. J. Aromather. 16, 57-62.

Orafidiya, L.O., Kaloustian, J., Agbani, E.O., Portugal, H., Abudu, A., Igbeneghu, O.A., Abou, L., Mikail, C., Ivaldi, B., Adetogun, G.A., 2007. A study of the composition, autoxidation and antimicrobial activity of the leaf essential oil of Ocimum gratissimum Linn against Helicobacter pylori and Neisseria sicca. Int. J. Essent. Oil Ther. 1, 103-109.

Owokotomo, I.A., Ekundayo, O., Dina, O., 2012. Ocimum gratissimum: the brine shrimps lethality of a new chemotype grown in South Western Nigeria. Glob. J. Sci. Front. Res. Chem. 12(6), 45-50.

Paula-Freire, L.I.V., Molska, G.R., Andersen, M.L., Carlini, E.L.A., 2016.Ocimum gratissimum essential oil and its isolated compounds (eugenol and myrcene) reduce neuropathic pain in mice. Planta Med. 82(03), 211-216.

Prabhu, K.S., Lobo, R., Shirwaikar, A.A., Shirwaikar, A., 2009. Ocimum gratissimum: A review of its chemical, pharmacological and ethnomedicinal properties. Open Complement. Med. J. 1, 1-15.

Sainsbury, M., Sofowora, E.A., 1971. Essential oil from the leaves and inflorescence of Ocimum gratissimum. Phytochemistry 10, 3309-3310.

Saliu, B.K., Usman, L.A., Sani, A., Muhammad, N.O., Akolade, J.O., 2011. Chemical composition and anti bacterial (oral isolates) activity of leaf essential oil of Ocimum gratissimum L. grown in North Central Nigeria. Int. J. Curr. Res. 33(3), 022-028.

Sofowora, E.A., 1970. A study of the variations in the essential oils of cultivated Ocimum gratissimum. Planta Med. 18, 173-176.

Sulistiarini, D., Oyen, L.P.A., Nguyen, X.D., 1999. Ocimum gratissimum L. In: Plant Resources of South-East Asia. No. 1 9: Essential Oils Plants. Prosea Foundation, Bogor, Indonesia, 140-142.

Unnithan, C.R., Sushen, U., 2017. Chemical composition of Ocimum gratissimum L by GC-MS analysis. Eur. J. Pharm. Med. Res. 4(06), 410-412.

Wabner, D., Geier, K., Hauck, D., 2006. For a deeper understanding of tea tree oil: Fresh is best-why we should only use fresh oil at any concentration. Int. J. Aromather. 16(3-4), 109-115.

White, R.L., Bugess, D.S., Manduru Mand Bosso, J.A., 1996. Comparison of three different in vitro methods of detecting synergy: Time-kill, chercherboard and E test. Antimicrob. Agents Chemother. 40, 1914-1918.