ISOLATION OF β-ASARON COMPOUND FROM SWEET . - WordPress

See discussions, stats, and author profiles for this publication at: ISOLATION OF β-ASARON COMPOUND FROMSWEET FLAG RHIZOME (Acorus calamus L.)USING CHROMATOTRON METHODSConference Paper · May 2016READS93 authors, including:Ahmad NajibUniversitas Muslim Indonesia17 PUBLICATIONS 8 CITATIONSSEE PROFILEAll in-text references underlined in blue are linked to publications on ResearchGate,letting you access and read them immediately.Available from: Ahmad NajibRetrieved on: 27 June 2016

Proc. of The Fourth Intl. Conf. On Advances in Applied Science and Environmental Technology - ASET 2016Copyright Institute of Research Engineers and Doctors, USA .All rights reserved.ISBN: 978-1-63248-097-2 doi: 10.15224/ 978-1-63248-097-2-18ISOLATION OF β-ASARON COMPOUND FROM SWEETFLAG RHIZOME (Acorus calamus L.) USINGCHROMATOTRON METHODSSuci Noviyanah Ansary1, Virsa Handayani2 and Ahmad Najib1*Abstract— Research on isolation of βasaron compound from Sweet Flag rhizomeAcorus calamus L. The aims of this research areto isolate and to identify mayor compound βasaronfrom Sweet Flag rhizome Acoruscalamus L. Sample from 150 gram driedrhizome, mascerated using n-hexane producing4,22 grams thick extract. Isolation of n-hexaneextract Sweet Flag rhizome Acorus calamus L.carried out by chromatotron methods using acomparison of eluen n-hexane : etil acetat (9:1).The isolate identified by UV-Vis spectroscopy,IR spectroscopy, and Gas ChromatographyMass Spectro. From the data result show thatisolate is β-asaron.II.Keywords— Acorus calamus L., β-asaron,chromatotron, isolation, Sweet Flag rhizomeI.MATERIAL AND METHODSThe sweet flag rhizomes (Acorus calamusL.) were obtained from the Bulukumba District,South Sulawesi Province - Indonesia.A. Sample ExtractionSample extraction by maceration using nhexane solvent. Sweet flag rhizome powder(Acorus Calamus L.) macerated with n-hexanesolvent. Maceration carried out for 3 days in aclosed container and protected from light,stirring periodically. Extract the results ofmaceration (maserat) separated by the pulp,pulp macerated with n-hexane solvent.Maceration done 3 times. The filtrate obtainedwas collected further concentrated by using arotary evaporator and the obtained extract nhexane.B. Thin Layer Chromatography ProfileN-hexane extract of Acorus Calamus L.reconstituted with the solvent n-hexane andthen spotted on TLC plates berukuruan 1 x 7cm by using a capillary tube. Each plate hasspotted insert into the chamber containing theeluent according to various comparison. Onceeluted, the plates are removed from thechamber and aerated until the eluentevaporates. Then the resulting chromatogramsprofile observed in the visible UV 254 nm and366 nm UV. Spotting obtained the observedand calculated nillai its Rf.C. Isolation of β-asaron by ChromatotronMethodsChromatotron plate with the stationary phasesilica gel 60 PF254 and 2 mm thick dried in anoven for 5 minutes at 50 C. The plates werethen placed in the chamber and then the motoris turned on and the plate wetted with n-hexanestream. Eluent flow is stopped immediatelyafter the first drops of n-hexane out. Thesample chamber is inserted into the hole withINTRODUCTIONSweet Flag rhizomes (Acorus Calamus L.)were included in Acoraceae tribe, known as amedicinal plant in India, USA, and Indonesia.Dringo (Acorus Calamus L.) can be used as asingle agent or as a mixture of medicinal herbs [8].The main chemical components (majorcompound) contained in this plant is en,humulen, methyl-eugenol, elemicine, cis-0cimen[6].Previous research by Ari Wahyuni (2011)reported that the main active compounds in Dringois β-asaron which are generally found in therhizome, but are also found in the leaves.Research on the chemical content andbiological activity of the plant Acorus Calamusalso been done that activity anthelmintic of theethanol extract of Acorus Calamus who grew up inSouth Africa, antifungal, antioxidant, inhibition ikandantioxidants,antihiperlipidemia and antibacterial [4].1Phytochemistry Division,Pharmacognosy DivisionFaculty of Pharmacy, Universitas Muslim Indonesia, Jl.Urip Sumiharjo KM 5, Makassar, Indonesia21

Proc. of The Fourth Intl. Conf. On Advances in Applied Science and Environmental Technology - ASET 2016Copyright Institute of Research Engineers and Doctors, USA .All rights reserved.ISBN: 978-1-63248-097-2 doi: 10.15224/ 978-1-63248-097-2-18the aid of a pipette and applied to the plateafter the n-hexane had not come out.D. Purificationa. Multi Eluen MethodsIsolates were then tested purity by usingsome variation of the eluent is based ondifferent levels of polarity. Multi eluentmethod using TLC plates 60 F254 with asize of 1 x 7 cm. Single sighting spotsindicating that the compound of isolatesobtained a single chemical components.b. Two Dimension MethodsIsolates obtained can also test the purityby using 2-dimensional manner isolatesditotol on the plates TLC 60 F254 with asize of 5 x 5 cm, then eluted using eluentis different to the first direction and thesecond direction, the elution process thelatter is done by 90ᴼ rotating plate. Thenthe chromatogram profiles were observedin the UV 254 nm and 366 nm.III.RESULTS AND DISCUSSIONSweet flag rhizome (Acorus Calamus L.) weretaken in Bulukumba District, South SulawesiProvince. Sweet flag plant Acorus Calamus L.intact and still fresh directly determined in theLaboratory of Pharmacognosy Faculty of PharmacyUMI-Phytochemistry. The result of determinationshows the type of plant Acorus Calamus L. fromAcoraceae tribe.Simplicia rhizomes Acorus Calamus L. thenextracted using maceration method. The extractionprocess is carried out using n-hexane solvent.Wherein the solvent is chosen because it has theability to attract non-polar compounds.TABLE 1. Results yield of n-hexane extract of thesweet flag rhizome (Acorus calamus L.)Solventn-HeksanSampleWeight(g)Quantityof Solvent(ml)ExtractWeight(g)YieldExtract(%)150 g2500 ml4,22 g3,55%N-hexane extracts obtained, then in profileTLC using eluent n-hexane: ethyl acetate in a ratioof 8: 2 and 9: 1. Of the TLC profile, a goodseparation of the stains found on the plates using aratio of 9:1.E. Identification of Pure Isolatesa. IdentifcationbyUV-VisspectrophotometryPure isolates then identified using UV-Visspectrophotometer. Compounds dissolvedin methanol pa then snippets put intocuvette (sample compartments) that differbetween monochromator and detector,resulting spectrum will be recorded on arecorder (Nurdin, 2014).b. Identification by IR spectrophotometryPure isolates followed by identificationusing infrared spectrophotometry byplacing the footage as a thin film betweentwo layers of transparent sodium chloride,then placed in infrared light betweenmonochromator with a detector, thenrecorded on a recording device (Nurdin,2014).c. Identification by GC-MSIsolates obtained from the purification ofthe next sample is placed on a mobilephase will bring the sample passedthrough the stationary phase, so that mostof the samples will be more likely to stickto the sample stationary and movinglonger than the other components so thateach component will be out on thestationary phase. The results are recordedon the recording device (Pavia , 2006).Figure 1. TLC profile of n-hexane extract of thesweet flag rhizome (Acorus calamus L.)Description:Stationary Phase: Silica Gel 60 F254Mobile Phase : n-hexane: ethyl acetatePlate Size: 1X7 cm(a) Appear spotting UV254(b) Appear spotting UV366Isolation of β-asaron extracts of n-hexanewas conducted using chromatotron, with silica gel60 PF254 adsorbent as a stationary phase wherechromatotron used plate thickness of 1 mm, using2

Proc. of The Fourth Intl. Conf. On Advances in Applied Science and Environmental Technology - ASET 2016Copyright Institute of Research Engineers and Doctors, USA .All rights reserved.ISBN: 978-1-63248-097-2 doi: 10.15224/ 978-1-63248-097-2-18the eluent n-hexane:ethyl acetate as the mobilephase with a ratio of 9: 4Biru0,54UnguTABLE 3. Results of Rf values and color patcheson the two-dimensional chromatography isolatessweet flag rhizomes (Acorus calamus L.)DirectionElutionArah 1n-Heksan :etil asetat(8:2)Arah 2n-Heksan :etil asetat(9:1)Figure 2. Instrument Chromatotron (Source:Personal Documentation)In the radial thin layer chromatography ischromatotron, obtained 65 vials were combinedinto 3 fractions. TLC profiles performed on eachfraction using the eluent n-hexane:ethyl acetate 9:1.UV 254ValueSpotRfColourValueRfUV ermore, the identification of thesingle isolates, using a spectrophotometer UV-Vis,IR Spectrophotometer, and GC-MS.Spektro data interpretation UV-Visibleshow maximum absorption at a wavelength of209.1 nm and 2,968 nm absorption is highest.Figure 3. Profile fraction of the isolatedchromatotron n-hexane extract of the Sweet flagrhizome (Acorus calamus L.)Description:Stationary Phase: Silica Gel 60 F254Mobile Phase: n-hexane: ethyl acetate (9:1)Plate Size: 1X7 cm(a) Appear spotting UV254(b) Appear spotting UV366Figure 4. Diagram UV-Vis spectrophotometry dataisolates the n-hexane extract of the sweet flagrhizome (Acorus calamus L.)SubsequentanalysisusingIRspectrophotometer to determine the functionalgroup of compounds that are in the isolates (Figure5). IR spectrophotometry of observation, theabsorption at 2959; 2927; 2873 cm-1 indicate thepresence of the CH stretching vibration of aromaticand CH2; Aliphatic CH3.After the testing isolates with a singlemulti TLC eluent systems and two-dimensional.TABLE 2. Results of Rf values and color patcheson multi chromatography eluent isolates sweet flagrhizomes (Acorus calamus L.)Mobile Phase(Eluen)Aseton:EtOAc6:4UV 254ValueSpotRfColour0,87BiruThe presence of C-O bond is shown in a sharp peakabsorption spectrum at wave numbers 1246 and1292 cm-1. Absorption band at 886 cm-1 indicate agroup C C (cis). Absorption band at 1464 cm-1 isthe vibration of the C C in the presence of thearomatic ring system.UV 366ValueSpotRfColour0,87Ungu3