Essential oil profiling of Centella asiatica (L.) Urb. a medicinally important herb

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1 South Indian Journal Of Biological Sciences 2016; 2(1); ONLINE ISSN: Research Article Essential oil profiling of Centella asiatica (L.) Urb. a medicinally important herb Santhi C. Francis 1, Thomas MT 1, * 1 St. Thomas College (Autonomous), Thrissur, Kerala , India * Corresponding author: Thomas MT; E mail: thomastbgri@gmail.com Received 1 June 2015; Revised 18 September 2015; Accepted 30 September 2015; Published 2 January 2016 Abstract The Centella asiatica is known as Gotukola. The essential oil is extracted by Clevenger apparatus. The oil yield from whole plant C. asiatica is 0.8 ml in 1 Kg. The essential oil is analyzed by GC MS. There are 43 volatile compounds are identified from essential oil of C. asiatica. P Cymene (44%) is the predominant compound in essential oil of C. asiatica. Keywords: Centella asiatica, Essential oil, GC MS analysis 1. Introduction Centella asiatica is a perennial herbaceous plant having pantropical distribution. This species is well known for its therapeutic and cosmetic applications. The plant is used as a major ingredient of formulations of Ayurveda, Siddha and Unani systems of medicine, and also in Homoeopathy along with modern medicine. In Indian pharmacopoeia, the plant is listed as useful for the treatment of skin malconditions such as leprosy, lupus, varicose veins, ulcers, eczema, psoriasis and wounds in addition to treatment of diarrhoea, dysentery, fever, amenorrhea and female genito urinary tract diseases, filariasis and tumour (Inamdar et al., 1996; Jain et al., 2007). It is also used as a brain tonic for the mentally retarded (Jain et al., 2007; Shukla et al., 1999; Liu et al., 2008). Centella asiatica contains several bioactive constituents, of which the most important are the triterpinoid saponins such as asiaticoside, madecassoside and centelloside. In addition, the species possesses other components including flavonoids, tannins, phytosterols, aminoacids and sugars. A large store of research findings are available highlighting the potential therapeutic effects of C. asiaitca against a variety of human ailments. Pharmacological studies revealed its usefulness in inducing human collagen 1 synthesis (Lee et al., 2006), accelerating nerve regeneration (Soumyanath et al., 2005), cardio protection (Pragada et al., 2004), improving age related neurological antioxidant status (Subathra et al., 2005), protection of critical neurons (Lee et al., 2000), treatment of venous hypertension (Incandela et al., 2001) and elevation of antioxidant level (Shukla et al., 1999) etc. The plant contain essential oil in which terpenoidal constituents such as β caryophyllene, β cymene, trans β farnesene, germacrene D, α terpineol, limonene, and linaloolwere identified. Among 169

2 these components, α terpineol is known for myorelaxant and antispasmodic effects. Linalool, the main compound of this oil, is a very important substance used in foodstuffs as additive and in pharmacology for its different properties. Another terpenoid, farnesol, have shown anti cancer effects and antibacterial activity in several different studies. Among the hydrocarbon, some compounds such as β elemene, limonene and β cymene are important compounds finding application in fragrance, pharmaceutical and agrochemical fields. Limonene has also been studied for its possible anti cancer activity. In the present study, detailed chemical profiling of the essential oil of C. asiatica occurring in the Thrissur region of Kerala was carried out. 2. Materials and methods 2.1. Collection of plant materials The plant Centella asiatica was collected from Chettupuzha, Thrissur Dist. Fresh plants were used for oil extraction. Oil extracted by hydrodistillation using Clevenger apparatus. The heating and condensation process repeated continuously for four hours. The essential oils obtained were dried over anhydrous sodium sulphate and stored in refrigerator at 4 o C till further analysis. Gas chromatograph mass spectrometry analysis was done on an Agilent 6890N Network GC System equipped with HP 5 (5% phenyl methyl poly siloxane, 30m x 0.32mm x 0.25 μm capillary columns) coupled with Varian CP 3800 GC SATURN 2200 ion trap mass detector. 1.0 μl of essential oil (1:10 dilution in diethyl ether) was injected under split mode (1:1 ratio v/v), with helium as the carrier gas at 1.4 ml/ min constant flow mode. The temperature programme for the analysis of the oils were, injector temperature 220 o C, oven temperature 60 o C (hold time 2 min) to 250 o C (3 o C/min) and interface temperature 290 o C. Mass spectra: Electron Impact (EI + ) mode, 70 ev and ion source temperature 250 o C. The essential oil constituents were identified by comparison of the relative retention indices calculated with respect to homologous of n alkanes (C6 C30, Aldrich Chem. Co. Inc.) (Van den Dool and Kratz 1963), MS library search (WILEY 275) and comparison of mass spectrum reported in the literature (Adams 2007). 3. Results and discussion The essential oil yield of Centella asiatica was 0.05% (v/w). The gas chromatograph of the essential oil of C. asiatica is presented in figure 1. The list of compounds identified and the percentage composition is shown in Table 1. Forty three volatile compounds comprising 93.3% of the total oil constituents were identified from Centella asiatica. Monoterpenoids were the major class of volatile chemicals (51.4%), with p cymene as the predominant compound (44.0%). Sesquiterpenoids consisted of 41.9% of the total compounds identified. Muurola 4,10(14) dien 1 ol (6.5%), elemene (6.2%) and E caryophyllene (5.2%) were the major sesquiterpenoids present in the oil. Characteristic aroma compounds such as 2 carene, limonene, myrtenol and carvone may be responsible for the peculiar aroma of the plant. Fig. 1. Gas chromatogram of the essential oil of Centella asiatica. 170

3 Table 1. Composition of the essential oil of Centella asiatica. Retention time Name of compound RRI Cal RRI Lit Percentage 6.4 α Thujene α Pinene ng 8.2 β Pinene Carene p Cymene Limonene p Menth,3,8 diene ng 12.6 Terpinolene ng 14.5 Allo ocimene ng 15.9 Terpineol ng 16.5 Meta cymen 8 ol Cis dihydro carvone Verbenone ng 18.1 Trans carveol Carvone ng 19.9 Trans myrtenol p Cymen 7 ol Myrtenyl acetate Neo iso verbenol acetate Elemene E Caryophyllene Guaiene ng ,9 Guaiadiene ng 27.8 Cis muurola 3,5 diene α Humulene Dauca 5,8 diene Trans cadina 1,6,4 diene Amorpha 4,7 (11) diene Aristolochene Germacrene B Spathulenol Caryophyllene oxide Globulol ng 34.0 Humulene epoxide II epi Eudesmol epi Cubenol Muurola 4,10(14) dien 1 b ol Valerianol epi α Eudesmol Elemol acetate Z α trans Bergamotol ,α 11 Elemodiol ng RRI Cal: Relative retention index calculated on HP 5 column, with respect to homologous of n alkanes (C6 C30, Aldrich Chem. Co. Inc.). RRI Lit: Relative retention index from literature (Adams 2007). ng: negligible (>0.1%) 171

4 Ali (2008) reported 23 compounds and α pinene, α humulene are same compounds seen in essential oil of C. asiatica. Oyedeji and Afolayan (2005) identified 40 compounds and they reported that the oil contain monoterpenoid hydrocarbon 20.20%, oxygenated monoterpenoid is 5.46%, sesquiterpenoid hydrocarbon 68.89% and oxygenated sesquiterpenoid 3.9%. In the present study monoterpenoid hydrocarbon is 49%, oxygenated monoterpenoid is 2.4%, sesquiterpenoid hydrocarbon is 26.9% and oxygenated sesquiterpenoid is 15%. Kanchan and Preeti (2013) reported 40 compounds. Humulene epoxide II, caryophyllene oxide, Germacrene B, α humulene, δ Terpinolene, limonene, p Cymene, β pinene, α pinene, α Thujene etc are same. As like that Supawan and Patchanee (2014) identified 87 compounds and similar compounds are Humulene epoxide II, caryophyllene oxide, Germacrene B, α humulene, δ Terpinolene, limonene, p Cymene, β pinene, α pinene, α Thujene etc. From Malaysia, Ali (2008) found 23 compounds, in essential oil of C. asiatica, from South Africa, Oyedeji and Afolayan (2005) found 40, from Taiwan, Chou (2005) reported 19, from different habitats of Nepal, and from India, Wong and Tan (1994) reported 41 compounds from the essential oil of C. asiatica. 4. Conclusion Essential oil yield of Centella asiatica is 0.08%. There are 43 compounds identified in the essential oil. Major compounds in essential oil are p cymene (44.0%), Muurola 4,10 (14) dien 1 α ol (6.5%), α elemene (6.2%) and E caryophyllene (5.2%), α Humulene (4%). Conflict of interest statement We declare that we have no conflict of interest. Acknowledgements The authors are grateful Dr. P. O. Jenson, Principal, St. Thomas College (Autonomous), Thrissur, Kerala for the facilities and to Dr. K. B. Ramesh Kumar, Scientist, JNTBGRI, Palode, Thiruvanandapuram, for the help rendered in interpreting the GCMS results. References 1. Adams RP. (2007). Identification of Essential Oil Components by Gas Chromatography/Mass Spectrometry. 4th edn. Allured Publ. Corp., Carol Stream. IL. 2. Ali MSM. (2008). Analysis of phenolics and other phytochemicals in selected Malaysian traditional vegetables and their activities in vitro. PhD Thesis. University of Glasgow. UK. 3. Chou YK. (2005). Studies on the essential oil constituent from the Hydrocotyle Sibthorpioides Lam. and biological assays. Master s Thesis. Chung Yuan Christian University. Taiwan. 4. Inamdar PK, Yoele RD, Ghogare AB, De Souza NJ. (1996). Determination of biologically active constituents in Centella asiatica. Journal of Chromatgraphy A, 742, Incandela L, Belcaro G, de Sanctis MT, Cesarone MR, Griffin M, Ippolito E. (2001). Total triterpenic fraction of Centella asiatica in the treatment of venous hypertension: a clinical prospective randomised trial using a combined microcirculatory model. Angiology, 52(2), S61 S

5 6. Jain P, Guiquing K, Chuanxun Y, Beibei Z, Risheng J, Yuan Y. (2007). Separation and determination of madecassic acid extracts of Centella asiatica using HPTLC with β cyclodextrin as mobile phase additive. Chinese Journal of Chromatography, 25, Kanchan J, Preeti C. (2013). Therapeutic Efficiency of Centella asiatica (L.) Urb. An underutilized green leafy vegetable: an overview. International Journal of Pharma and Biosciences, 4(1), Lee J, Jung E, Kim Y, Park J, Hong S, Kim J, Hyun C, Kim YS, Park D. (2006). Asiaticoside induces human collagen I synthesis through TGF beta receptor I kinase (Tbeta RI Kinase) independent Smad signalling. Planta Medica, 72(4), Liu M, Dai Y, Li Y, Luo Y, Huang F, Gong Z. (2008). Madecassoside extracted from Centella asiatica herbs facilitates burn wound healing in mice. Planta Medica, 74, Oyedeji OA, Afolayan AJ. Chemical composition and antibacterial activity of the essential oil of Centella asiatica growing in South Africa. Pharmaceutical Biology, 43(3), Pragada RR, Veeravalli KK, Chowdary KPR, Routhu KV. (2004). Cardioprotective activity of Hydrocotyle asiatica L. in ischemia reperfusion induced myocardial infraction in rats. Journal of Ethnopharmacology, 93(1), Shukla A, Rasik AM, Jain GK, Shankar R, Kulshrestha DK, Dhawan BN. (1999). In vitro and in vivo wound healing activity of asiaticoside isolated from Centella asiatica. Journal of Ethnopharmacology, 65, Soumyanath A, Zhong YP, Gold SA, Yu X, Koop DR, Bourdette D, Gold BG. (2005). Centella asiatica accelerates nerve regeneration upon oral administration and contains multiple active fractions increasing neurite elongation in vitro. Journal of Pharmacy and Pharmacology, 57(9), Subathra M, Shila S, Devi MA, Panneerselvam C. (2005). Emerging role of Centella asiatica in improving age related neurological antioxidant status. Experimental Gerontology, 57(9), Supawan R, Patchanee Y. (2014). Antibacterial, antioxidant and chemical profile of Centella asiatica. Biomedical and Pharmacology Journal, 7(2), Van den Dool H, Kratz PD. (1963). A generalization of the retention index system including linear temperature programmed gas liquid partition chromatography. Chromatography, 11, Wong KC, Tan GL. (1994). Essential oil of Centella asiatica (L.) Urb. Journal of Essential oil Research, 6(3),

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