J. Nepal Chem. S0c., Vol 39, 2018 Isolation and Characterization of Essential Oil from Cymbopogon citrates Grown in Nepal ‘Shanta Pokhrel” and Sanjay Kumar Yadav Department of Chemistry, Tri-Chandra Multiple Campus, Tabhuwan University, Kathmand, Nepal Email: shentabhattarca2014@gmail.com Abstract In this work essential oil was obtained by hydro-distillaion from air dried leaves of Cmbopogon citrates using Clevenger apparatus and analyzed by Gas chromatography-mass spectrometry (GC-MS) technique. Twenty three chentcal components were identified in the escental ail of Cmbopogan citratus leaves. Among them geranial (tral-a) 32.51% neral (etral-p) 28 38%and myrcene 14.23 %ewere found ‘as major components. The prekimtnary phytochenscal analy.as of the methanolic extract of Cymbopogon citrates leaves showed that it has alkaloids, tamins, carbohydrates and flavonoids whereas negative results for phenols. Keywords: Cymbopogon citrates, Essential ail, CQtval-a, Cltral-p, GC-MS Myrcene Introduction Plants having essential ils ee significantly important in metlicel practice. The chemical composition of essential oil is complex and may contain hundreds of components. Its @ mixture of fiegrent volatile substances such as monoterpenes, semputerpenes, emomatic compounds, and their denivatives. Essential oils have been used os antimicrobial, ent-inflammatory, sedative, expectorens, and diaphoretics ete ‘There are numerous mesicinal plants possess essential oils in their tissues such as sage (Salvia offcinais L), ewcalyptus species, peppemuint (Mentha pipenita L.), Scotch pine, elecampane (Bula heleton L), linden (Tika cordata Mi, garden heliohope (Valeriana officinalis L.), etc! Lemongrass, Chmbepogon citratus, is « perennial medicinal plant belonging to family Geremineae, ‘and itis distabuted worldwide in tropical and subtopicel areas of Asia, Afice, and America®, Some species (particularly Grmbopogan citratus) ae commonly cultivated as culinary and medicinal heibs ‘because of their scent resembling that of lemons (citrus lemon). Common names inchudes lemon gras, suibed wite grass, silky heeds, citoelle gras, fever grass, tangled hewba or gavatichehapati among anyother Lemongrass is widely used as culinary heib in Asian cuisines and also as medicinal heyb in Inia It das @ subtle citrus flavour and can be dried and povrdezed or used fresh, It is commonly used in tas, soups, acl cures, It is also stele for the use with poultry fish, beef and sea food. It soften used as a tea in African countries such as Togo, South Eastem Ghana Volta region and the democratic republic of * Corresponding author -70-J. Nepal Chem. S0c., Vol 39, 2018 Congo and latin American counties such es Mexico’. Lemongrass oil is used as pesticides and esewative, Research shows the lemongrass oil has antifungal properties. Despite its ability to repel some insects such as mosquitoes, its il is commonly used as ‘Inve’ to attract honeybees' Lemon grass Chmbopogan nardis and hmbopogan winteiamus grow to about 2m (66 andl have ragenta colowed base stems, These species are used in the production of soaps, insect repellant (especially mosquitoes) in insect says and cendles and in aromatherapy. The prieipal chemical ‘constituents ie. geraniol and cittonellol of citonella, are the antiseptics”. It is used as a pesticides and ‘preservatives, is put on the ancient plant leaf manuscripts found in Nepal and India preservative® It is also ‘wed as an edition fo tea, and in pepazaons such as kadh, which is «traditional herbal brew used geist coughs, cold, ete, It has medicinal properties too and is used widely in ayurvedic medicine. Its supposed to help with relieving cough and nasi congestion It shows antiordent, ant-cencer roperties snd an-inlanunatory property? Herbal droge ave been wsed worldwide during the last few decades as evidenced by mpl growing global and national mazkets of heal druge!®. Several studies indicated that appwoximstely 3% of drugs ‘produced in the developed countries are derived from plants”. The present research was carried out to evaluate phytoconsttuents of methanolic extact of GymBopogan citrate: leaves and ivestigete the composition of it extacted essential ol wa Gas chromatography-massspectiomety (GC-MB) technique Experimental Methods ‘Meterials and Sample Preparation ‘The fiesh leaves of Cymbopogon citrahus were collected from the Nepal Agricultual Research CComeil Lalitpur, Nepel. Fesh and dried leaves of were subjected to hyto-dstilation for 8, using Clevenger type appuetis, according to method recommended by Guenther °. The extacted essential cls were dried with anhyezous sodium sulphate and stored in sealed eubet at ow temperate before analysis ‘The exacted oil was analyzed by GC-MS using Varian 240 GC-MS system equipped with VF-S fused capillary colurm (30m x 0.25 mm, ie. film thickness 0.25 nn) oven temperature was 50-180 °C ata rte of 5 “OMmin, tansfer line temperature 250 °C, cemier ges was helium witha flow rate of 1 mL/min, split ratio 1:20, ionization energy 70 eV, and mass range 35-390 amu. The constituents of the oil were identified by companson of their mass spect with those of a computer Library or with authentic compounds Phytochemical screening For phytochemical sereening, the plant samples were crushed and blended into into smaller pieces 200 g of powdered Cymbopogon citratus samples (leaf) were taken into 2000 ml. capacity conical las 1000 mL. methanol was poured to the sample. The conical flask containing the mixtues was placed ona shaker for 24 hows and filtered the content through muslin cloth. The filhate was then filtered again ‘using suction presswe with the aid of a vacuum pump. The filtered extact was concentrated using the rolary evaporator equipment then dried on en evaporating dish at a temperature of 50-60 °C to a semi- solid form A sticky semi-solid greenish substance obtained which was subjected for phytochemical onJ. Nepal Chem. S0c., Vol 39, 2018 screening using standard rocedwes"™"* The following qualitative tests for both the metabolites were done as follows: Test for alkaloids About 10 mg of exact was taken and few diops of Wagner's reagent were added and the formation ofa reddish brown jrecipitate indicates the presence of alkaloids Test for Flavonoids Shinoda Test: 10 mg of extact was added to pinch of magnesium tunings and 1-2 drope of concentrated hythochloric acid was added. Fomuation of pink color indicates the presence of Flavanoids. Lead acetate test: 10 mg of extract was taken and few drops of 10% lead acetate solution was added. Appearance of yellow colour precipitate indicates the presence of flavonoids. Test for Phenols and Tannins Lead acetate test: 10 mg of exact was taken and 0.5 mL. of 1% led acetate solution was added and the formation of precipitate indicates the presence of tannins and phenolic compound Femic chloride test: Five mg of extact was taken and 0.5 mL of 5% ferric chloride was added. The development of dark bluish black color indicates the presence of tannins Sodium hydioxide test Five mg of extract was dissolved in 0.5 ml of 20% sulphuric acid solution. Followed by addition of few dope of aqueous sodium hydroxide solution, it tune blue which indicates the presence of phenols Test for Carbohydrates Febling’s test: Five ml of Fehling’s solution was added to 0.5 mg of extract end boiled in a water bath ‘The formation of yellow or red precipitate indicates the presence of reducing power. Benedict's test Five ml of Benedict’s solution was added to 0.5 mg of exttact and boiled in water ‘bath. The appearance of red or yellow or green jecipitate indicates the yresence of reducing sugers Results and Discussion A pale yellow essential oil with yields of 0.67% (on fiesh weight basis) was obtained from fiesh lemongrass plant. This vesult agrees with some works who reported that oil content ofa nomnal cut should average 0.25.0.5%, but with good management and selected strains could be yielded up to 0.66- ‘ool ‘Twenty three components were identified in the essential oil of fiesh air dried C. citvatus leaves by different drying methods, which represented 100 % of the oil components. The chemical constituents of coils are resented in Table 1. The components are listed in onder of their retention time on the VF-S column, na:J. Nepal Chem. S0c., Vol 39, 2018 Table 1: Essential eil components of Cymbopogan citratus Peak® B Tine ‘Area Wane T 1427 TsO Hep Sen D-one a THIS? 31509 Ocimene <(E)bete> 3 104 341462 Linaloal é 19594 T0733 RT 19.592 7 Tait WOE T52__| Sabinene hyciate Stans > 3 Wass iS __ | Octe-7, 4dienol 9 WaT T__| Thajone bete> ig O34__| None-OE,62)-dienal i 00660 036___| Gaysanthemel 7 1731067 134 | Veibenal a BOSS TS) | Rove Fuan oxide 4 2310227 DOT | Teogeranial 5 1013333 O91 | Bthenal 16 26:16 Tas T2__ [Oct Tenol3,T-aimethyi> 17 26588 31667250, 238 | Neal ig aia BAIT 385__| Geraniol @ 27934 38505521 3452 | Gerenial 20 30516 BI O25 __| Deca GE aB)-dien Tol a Da 250930 O25 | Unniee-(Sz)-enal 2 a TASES TH | Linaloclisobutyate 3B ATS 224109 020 | Exdesmol 11872191 100.00 ‘The major components of the essential oils were geranial (34.52%), neral (28.38%) and myreene (423%) in oils extracted from air dried lemongrass leaves. The quality of lemongress is generally determined by its citral content. Citral is combination of bioactive isomexs geranial and neal According to Ref” the lemongrass essential oil is urually made up of cit at an average of 65 to 30% ‘This result showed 23 major components of the essential oil of lemongrass. Among 23 compounds, only 10 compounds (6-Methyihept-Sen-2-one, Myrcene, trans Sebinene hydrate, trans-Verbenol, Rose furan oxide, Isogeranial, Neral, Geraniol, Geranial and Linalool iscbutyrate) had a concentration of greater than 1%. This is in agreement with severe] research work, reported the vatiations of 20 to 50% of “73.J. Nepal Chem. S0c., Vol 39, 2018 ‘geranial and 30 to 40% of neral in lemongrass chemical composition’*"**. According to Schaneberg and. ‘han, the essential oil of lemongrass contains mainly geranial and neve”. Other isolated components, suchas P-myrcene, ocimene, f-cimene, linlool, cittonellal,citronellol, caryophyllene and f-pinene, were present as minor components reported in Ref ‘Table 2 shows the phytochemical constituents of methanolic exact of lemongrass leaf. The result shows the presence of alkaloids, tannin, flavonoids and caibohytrete whereas phenol is absent This ‘agrees with the findings of Umar etal." and Aina e¢ al” Table 2: Phytochenscal constituents of Cymbopogon citrates leaf Phyiocheniral Constituents | Leaf extract Gmetianol) ‘llaleide + Tain + Flavonoids + Plead ; Cabolyehate + Conclusions ‘The essential oil was successfully exhacted fiom the plant Cymbopogon citratus by means of hythodisilation method va clevenger type apparatus. The volatile constituents which is present in this lant was analyzed by the Gas Chuomatography-Mass Spectroscopy. Twenty tuee chemical components ere identified inthe essential oil of Cymbopogen citratus eaves and dried citrus leaves obtained by GC- (MS technique including geranial (cittal-a) 32.51%, neval (citval-f) 28.38% and myreene 14234 ‘major component. The result of phytochemical sereening of methanolic extract of Chmbopogon citratus leaves showed the presence of tannins, lavonoids and caibohyytrates whereas phenol is absent. Acknowledgements We would like to acknowledge Department of Plant Resouces, Thapathali, Kathmandu, Nepal for GC-MS analysis. The authors gratefully acknowledged the Department of Chemistry, Txi-Chandra ‘Multiple Cempus, Tribhuvan University, Kathmandu, Nepal for providing available facilities to conduct this work References 1. W.S. Soliman, S. Salaheldin, HM Amer, International Journal of Scientific & Engineering Research, 2017, 8, 630. A Alia, CRC Press, Taylor and Francis group, 2010, 12, 108. 3. V. Francisco, A. Figueisinha, B.M Neves, C. Garcia-Rodriguez, M.C. 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