ISSN: 2277- 7695 Chemical Fingerprint Of Essential Oil .

The Pharma Innovation Journal 2015; 3(12): 50-56ISSN: 2277- 7695TPI 2015; 3(12): 50-56 2015 TPIwww.thepharmajournal.comReceived: 02-01-2015Accepted: 16-01-2015Pressy P. PrakasiaDepartment of Botany, Universityof Kerala, Kariavattom campus,Thiruvananthapuram, Pin Code:695581, Kerala, India.Ashalatha S. NairDepartment of Botany, Universityof Kerala, Kariavattom campus,Thiruvananthapuram, Pin Code:695581, Kerala, India.Chemical fingerprint of essential oil components fromfresh leaves of Glycosmis pentaphylla (Retz.) CorreaPressy P. Prakasia, Ashalatha S. NairAbstractThe present research was undertaken to characterize the promising bioactive constituents of Glycosmispentaphylla leaves. The volatile oil was obtained by hydrodistillation and the components present wereanalyzed by gas chromatography-mass spectrometry. Elucidation on mass spectrum GC-MS wasconducted using the database of National Institute of Standard and technology (NIST). The GC-MSanalysis of essential oil of Glycosmis pentaphylla leaves resolved into sixty seven compoundsrepresenting 99.71% of essential oil. The active principles with their retention time, peak area, molecularformula, molecular weight, structure and category of the compound were predicted. Phytol (28.03%)was the dominant compound in the oil. From the present study, it is revealed that the oil present in leaf isvery rich in diverse phytochemicals. Most of the identified compounds are basically biological important.The results are in conformity with the tribal conviction for which they use as traditional medicine fordiverse bioactivities and curing of ailments.Keywords: Glycosmis pentaphylla, essential oil, GC-MS, Phytol, sesquiterpenes.Correspondence:Pressy P. PrakasiaResearch Scholar, Department ofBotany, University of Kerala,Kariavattom campus,Thiruvananthapuram, PinCode: 695581, Kerala, India.1. IntroductionThe therapeutic use of plants against critical human illnesses predates recorded history andrepresents the most significant direct antecedent to modern medicine [1]. Scientific research hasallowed discovering a wide range of active constituents, of which the most important as far ashealth is concerned are essential oils, alkaloids, glycosides or heterosides, mucilage and gums,and tannins. The active constituents specific to a particular species are characterized aschemical markers. Use of chemical markers is an effective tool to resolve problems instandardization of botanicals, using chemical fingerprinting and in chromatographic fingerprinting of botanicals to demarcate them on the basis of their chemotypes and geographicalorigin [2].Glycosmis pentaphylla, belonging to the Rutaceae family, is a shrub or small (1.5–5 m) tree,widely distributed, spanning from India, Malaysia and Southern China to the PhilippineIslands where it occurs in tropical forests at low altitudes. It has been used as folk medicine forthe treatment of fever, liver complaints, jaundice, cough, eczema, anaemia, diarrhoea, andrheumatism [3, 4]. Phytochemically speaking, Glycosmis pentaphylla were investigated on a fewoccasions. Most of the phytochemical work realized in this field resulted in the isolation ofhydrophobic alkaloids acridone, carbazoles, quinolones and quinazolones [5-8]. Studies showedthe extracts of G. pentaphylla having potent anthelmintic, antipyretic, hepatoprotective,antibacterial, antioxidant, antidiabetic and antinociceptive properties [9, 10]. Regarding thephytochemistry of essential oil of G. pentaphylla, the composition of essential oil was shown,and aliphatic ketones 2- tridecanone 6,10,14- trimethyl-2-pentadecanone were the majorcomponents identified [11]. Review of literature divulges that the essential oil compositionwithin the species may vary significantly and such differences in the composition of theiressential oil could be due to natural chemical variation called chemotype, which occur in thesecondary metabolism of plants and could possibly due to organ of the plant studied and alsoinduced by environmental factors such as soil type, altitude, sun exposure, rain and seasonalvariation besides the method of oil isolation [12]. As far the literature investigation couldascertain, the fresh leaves of Glycosmis pentaphylla were never examined regarding theirvolatile oil composition. Hence, the aim of the present study is to provide the first detail GCMS analysis of the volatile components of the fresh leaves of Glycosmis pentaphylla. 50

The Pharma Innovation Journal2. Materials and Methods2.1 Plant materialFresh leaves of Glycosmis pentaphylla were collected from itsnatural habitat from district of Thiruvananthapuram (Latitude8.54ºN and Longitude- 76.91ºE), Kerala, India, in January2014. The titled plant was botanically identified by Curator,department of Botany, University of Kerala and a voucherspecimen (KUBH 5858) has been deposited at the herbariumof Botany department, University of Kerala, India.2.2 Extraction of essential oilThe leaves were slightly washed to remove dust and otherphysical contaminants. The leaves were then reduced to asuitable size using electric blender and loaded them in theextraction flask. The essential oil was extracted byhydrodistillation for 8 h (60 g of sample in 500 mL of distilledwater) using Clevenger apparatus [13]. The essential oilobtained was separated from aqueous phase and stored insealed glass vial protected from the light at 4 ºC until analysis.The oil sample was subsequently analysed by GC-MS.2.3 GC-MS analysisThe analysis of the oil was performed using GC-MS (Model:GCMS- QP 2010, Shimadzu, Tokyo, Japan) equipped with aVF 5 ms fused silica capillary column of 30 m length, 0.25mm diameter and 0.25 mm film thickness. For GC-MSdetection, electron ionization energy of 70 eV was used. Thecarrier gas was Helium (99.99%) used ata constant flow rate of1.51 ml/min. Injector and mass transfer line temperature wereset at 2000 C and 2400 C respectively. The oven temperaturewas set from 70 to 2200 C at 100 C/min. Two µL of samplewas injected in a split mode with a scan range of 40-1000 m/z.The total running time of GC-MS was 31 min. The relativepercentage of the extract was expressed as percentage withpeak area normalization.2.4 Identification of the compoundsElucidation on mass spectrum GC-MS was conducted usingthe database of National Institute of Standard and technology(NIST). The spectrum of the unknown components wascompared with the spectrum of the known components storedin the NIST08 library source [14]. The name, molecular weightand molecular mass of the identified compounds were furtherconfirmed by comparison of their retention indices withliterature data. For quantitative analysis, compoundsconcentrations (as % content) were calculated by integratingtheir corresponding chromatographic peak areas.3. ResultsThe essential oil of fresh leaves of Glycosmis pentaphylla wasobtained by conventional hydro- distillation method using aClevenger apparatus. The hydrodistillation gave characteristicodour, golden yellow oil. In this study GC-MS fingerprintingof essential oil of fresh leaves of Glycosmis pentaphyllarevealed several peaks. The gas chromatogram shows therelative concentrations of various compounds getting eluted asa function of retention time (Figure 1). Identification of thecompounds was accomplished by comparing their massspectra and retention indices with those given in the literatureand those authentic samples. The active principles with theirretention time (RT), molecular formula, molecular weight(MW) and concentration (%) are presented in Table 1. Theyare listed by their order of retention times. The heights of thepeak indicate the relative concentrations of the compoundspresent in essential oil of Glycosmis pentaphylla leaves. Sixtyseven compounds were identified representing about 99.71%of the total oil’s compounds.The most prevailing major compounds were Phytol imethyl-4-vinyl(10.93%), 1,19-Eicosadiene (9.84%), 1,6-Cyclodecadiene, 63%),Caryophyllene oxide (4.32%), (-)-Spathulenol (3.92%) andBicyclogermacrene (3.55%)., and the minor compounds wereBicyclo[4.4.0]dec-1-ene, 2-isopropyl-5-methyl-9-methylene(2.11%), Epiglobulol (1.75%), (-)-Globulol ethyl-1-(1methylethyl)-(1S-cis)-(1.53%), 1H Indene, (1.33%), Ledol(1.33%), Cyclohexane ,1-ethenyl -1-methyl -2,4-bis 1.23%),Santolina triene (1.23%), Toluene (1.20%), Humulene(1.20%), gamma.-Elemene (1.18%), 2-Pyrrolidinone (1.10%)and Cyclohexane, hylidene)- (1.05%). The remaining compoundswere present in less than 1%.Chemical class of the detected volatile compounds aredisplayed in Table 2. The compounds were separated intomonoterpenes, sesquiterpenes, diterpenes, alkanes and alkenes,fatty acids and others. The most representative compoundswere sesquiterpenes (46.39%) followed by diterpenes(28.58%), fatty acids (10.61%), monoterpenes (5.81%) andalkane and alkene (3.83%). Major phytocompounds and itsbiological activities obtained through the GC-MS study ofleaves of Glycosmis pentaphylla are presented in Table 3. Thebiological activities listed are based on Dr. Duke’sphytochemical and ethnobotanical online databases by Dr. JimDuke of Agricultural Research Service/ USDA.4. DiscussionThe chemical and chromatographic fingerprints provideadequate information about the safety and credibility, withevidence for the product. A chemical profile of sixty sevencompounds was identified representing about 99.71% of thetotal oil’s compounds. Usually the major compounds mirrorthe biological activities of the essential oil from which it isextracted. Essential oil of G. pentaphylla composed mainly ofterpenes (monoterpenes, sesquiterpene and diterpene) and itsbiological activity may be attributed to its high concentration.However, the activity of major components may be modulatedby other minor components present in the oil.Among the identified compounds, the diterpene alcohol,Phytol, which is at hand in highest with 28.09% , is vital in thedispensation of glucose and can trigger enzymes within thebody that have strong positive effects on insulin level. Thismeans that Phytol in the human diet could perhaps helpreinstate the metabolic activities of those with type-2 diabetes.Phytol confirmed a strong antioxidant effect in vitro in itscapacity to remove hydroxyl radicals and nitric oxide as wellas to prevent the formation of thiobarbituric acid reactivesubstances (TBARS) [15]. Sesquiterpene biosynthesis seems tobe complex since the formation via either pathway (mevalonicor methylerythrytol) or a combination of both has beenreported [16]. Nevertheless, these appear to be ubiquitous inplant taxa and some insects, and are associated to the cytosol–mitochondria. Sesquiterpenes were reported to have antihyperlipidemic activity. 51

The Pharma Innovation JournalFig 1: GC-MS Chromatogram of essential oil of Glycosmis pentaphylla leavesTable 1: Phytocompounds identified in the essential oil of Glycosmis pentaphylla leaf by 370.471.2Name of the compoundRTToluene3- Hexen-1-ol, (Z).beta. – Pinene2-Pyrrolidinone, 1-methyl.beta.-Ocimene1,6- Octadien-3-ol, xenyl enylpyrazoleCyclohexane, e]azulene, o[5.2.0]nonane, 96.627.619.119.369.8611.5611.611.79 52 .280.290.70.130.110.27