solvent front, Some gesieral points should be ‘considered in terms ofthe extraction process, sich as the overall characteristics of the secondary rmetabolitex to be extracted (e.g. some glycosides are thermolabile oF pH-sensitive). Although the normal practice is to apply 2iitanOarO technique to obiain a ‘onide extract from a plant material, e.g. an acid-base shakeout to prepare an alkaloidal extract, beeause of Ue stnictural diversity within a given natural proxduet group and thelr possible special requitements, it is advisable 40 consult specific reviews, papers, and books in order to prevent the avoidable loss of desired bioactive metabolites caused by the tse of an Inappropriate extraction technique. The most simple ‘extraction processes employed may be classified as follows: extraction with organic solvents: percolation, maceration, and extraction using a Soxhlet apparatus: and extraction with water: infusion, decoction, and stear on The most popular method of extraction is to use a Liquid solvent at atmospheric pressure, possibly with the application of heat. Other methods include steam distillation, supercritical fluid extraction and the use of liquefied gases under moderate pressure, The ‘choice of method depends on the factors listed above as well asthe intrinsic advantages and disadvantages, of the procedures." ‘Supercritical Maid extraction ‘The process of separation of one component which is ‘extracting from matrix by using supercritical fluid ix known as supercritical fluid separation, Supercritical fluids showed property intermediate between those of the liquid and gaseous phases, for any substance it ix a condition above the critical temperature and pressure.SFE offers many advantages as follows: 1. It leads to lower solvent usage 2, Controllable selectivity 3. Cleaner extracts an! less thermal degradation as compared to conventional solvent extraction andl steam distillation methods, Super critical carbon dioxide (SCO2) - with its particularly attractive properties such as non toxicity, non-flammability, — non-corrosiveness, ‘chemical inertness, low critical temperature (304"K), riodetately low critical peessure (73 atin), easy availability, co-elfectiveness and environmental acceptability is the preferred solvent for many super critical extractions. Liquid carbon dioxide ix completely miscible with componests of essential some time, proteins, starches, mineral salts and water are insoluble in liquid carbon dioxide, Essential oils obtained by liguid carbon dioxide extraction are superior to that obtained through steam distillation and solvent extruction, Extraction of several natural products such as pyrethtins from chrysaetherwunn flower, essential oils from anise, caraway, clave, star anise, cinnamon and ginger are inereasingly done by this process," Solid phase extraction Solid phase extraction is process of separation of dissolve and suspended component from liquid ‘mixture by using another component in the mixture secording to their physical and chernieal peoperty.9 Distillation Distillation may be defined as separation of components of a mixture of (wo oF more liquids by Vite of difference in theie vapor pressure, There are Ahee systems of distillation + Hydio distillation © Hydio-steam distillation + Steam dst lation Hydro distillation-Hydra distillation is the oldest method being used for separation of essential oil. In this method plant material is contact with boiling water in a crude metallic distillation unit, ‘This process use principle of osmotic press principle of ostiotic pressure to diffuse oil from the oil glands ‘The essential ofl of a plant consists of muny compounds which generally boil between 150° to 300° C. The vapors pass through a coiled tube contained in a water bath and condensate is obtained at the bottom of the condenser tube. The disadvantages are that the heat is difficull to control and hence the rate of distillation is variable, Also the possibility exists for local overheating and “burning” of the charge which ean lead ta poorer quality oil. Hydro-steam distillation To overcome the drawback of water distillation, modifications in techniques was developed. In this technique plant material is supported on a perforated arid oF screen inserted at some distance above bottom of still, Water filled below the grid is heated which produce saturaied and wet steam; produced steam pass through plant material and vaporized essential oilReview Article Current Pharma Research ISSN: 2230-7842 CPR 2(3), 2012, 584-899. Isolation of Natural Products *Sharad Visht, Swati Chaturvedi. Department of Pharmaceutical Technology, Meerut Institute of Engineering and Technology, NH-S8, ‘Baghpat Bypass Crossing, Delhi-Haridwar Highway, Meerut-250005 (UP), India. Abstract Plant had been used as medicine in ancient, Now day’s Pharmaceutical companies start processing of medicinal and aromatic plants in their formulation by using extraction of active components. Extraction of plant ‘components likes volatile, Essential or ethereal oils and mixtures composed of volatile liquid and solid ‘compounds depend upon their composition and their boiling paint. Now days there are several processes. like distillation, enfleurage, maceration, expression, solvent extraction and fluid extraction are available for extraction of plant component. This review also summarizes the characters of phytoconstituents, choice of solvents, influence of solvents, extraction strategy, procedures for extraction of herbal drugs and treatment of «drug residue after extraction Key Words Secondary metabolites, Phytoconstituents, Enflurage. Introduction [Natural products are secondary metabolites wi are derived from. herb or animal sources. Natural products are chemical compound found in nature and they have pharmacological and biological activity Natural products are generally used in deug discovery and drug design. Separation of a single ‘molecular entity is very difficult from complex ‘mixtures contain fats, oils, alkaloid, tannins and glycoside. In 1803 the first alkaloid, nicotine and then morphine, strychnine, emetine and many others ‘were separated. This review compiles the recent literature with special focus on various approaches for extraction, This review also summarizes choice of solvent, interfering compound and strategies involve in extraction. ‘Isolation of Natural Products As time passes new separation techniques and analysis are being introduced to separate different nature of compounds like alkaloids, glycosides, steroids, saponins, tannins, flavonaids ete. Carbohydrates, fats, and proteins are considered more valuable as they are having dietetic importance.'"* Many starches and gums are used in pharmacy but lack any marked pharmacological action; are used as binder, viscosity builder and as hydrocolloid to increase the stability of emulsions and suspensions? "Corresponding Author: sharadvisht@ yahoo.com Afier GATT there is a great surge to find out immense potential of traditional and other herbal drugs to prevent and cure diseases. Several extraction, frictionation/separation and isolation methods are developed afier which isolation of the active moiety and their chemical examination is performed. Microwave extraction, sonication, lyophilization, spray drying and vacuum drying have also heen employed with good results. Generally the extraction is based on pharmacological activity rather than chemical nature of compounds. Further studies are carried out after the isolation of the active moiety to confirm the compound? The phytochemical investigation of a plant involves the selection, collection, identification and authentication, extraction of the plant material (first fractionation), fractionationseparation (second fractionation) and isolation (third fractionation) of the constituents, characterization of the isolated compounds and investigation of the biosynthetic pathways of particular compound, quantitative evaluations and pharmacological activities." Terms and Definitions The extraction of drug represents either solid-solid for liquid-liquid extraction, Some common terms used in plytochemsistry are given in Table | Principle Extraction processes for drugs can depends on the partition of component between solvent phase and solid residual and dependent on diffusion ofSawer o1096207 A new formulation of insecticide, repellent, and larvacide against Proceeding of The Intemational Sein om Chany 2008p. 595 588) mosquitoes from the waste product of Jeruk Nipis (Citrus aurantifolia) syrup industry Roekmi-ati K. Tjokronegoro', Sayuti Murad?, Sri Surjati Imron*, Hasyim Asnari! "Department of Chemistry, FMIPA, Unpad “Department of Biology, FMIPA, Unpad “Faculty of Pharmacy, Unpad Abstract ‘A new natural insecticide, repellent and lar vacde against Aedes aegypti mosquito, a veetoe for dengue fever, was successfully developed from the waste product of jerut mipis (Citrus auranijolia) collected fom the syrup industry in Kuningan . The compounds of the formula were obtained by hydrodisillation of the fruit pee for 20 hours, which produced the essential oil, followed by soxblet cextaction of the residue with ethyl acetate for 10 hours to obtain the non-volatile extract. The insecticide formulation was consisted ofa solution of 0.2E(v/v) of the essential oils containing 60" ‘of the active component I-limonene in ethanol, with Paraben as a conservant. By means of spraying, the insecticide resulted in 100% morality of the mosquitoes. The the main active component in the non-volatile extract was 2,3 dihydeo-benzofuran. The repellent was successfully formulated screams and stick repellent containing 30cE(whw) of the non-volatile extract with 215% 23 dihidro- benzofuran , as the main repellent compound, mised with the usual base compounds for repellents. ‘The repellent showed repellent activity for & hours. The larvacide was formulated as 10 grams granules in bags and as ethanol solution 10 ml_ , which consists of 1.3 grams of the non-volatile extract for 1 L of water. The larvacide formula resulted in 100 mortality of larvae in under 24 hours ‘The advantage of the new formula over those which are now commercially in circulation is tht itis not toxic to human, has pleasant ance, friendly to the environment, and it might be also cheaper. Keywords: Insecticide, repellent, larvacde, formulation, Cis aurantifolia Introduction Citrus peel contains plenty of bioactive compounds, including insecticidal, repellent and larvacidal compounds. I-limonene in the essential oil and the flavonoids in the non-volatile fractions of citrus peel ‘are attributable for its insecticidal and repellent activity against adult mosquitoes and the larvae The Citrus fruit (Citrus aurantifolia), locally known as jerk nipis, is the main ingredient for a beverage industry in Kuningan, West Jawa, and until recently the fruit peels which is the waste product has ‘not been utilized yet. The home industries use atleast 60 to 80 tons a week. The method of extracting the essential cil from the fruit peel has been investigated bby Kartika (2000). It has been shown that this method, ‘which was hydrodistllation of erushed fresh fruit peel ‘of Citrus nobis for 20 hours, produced the highest quantity of essential oil (0.54% viw). Puspitasari (2002) reported that an insecticide against Aedes ‘aegypti in C. nobilis essential oil is already known as ‘Hlimonene, as shown below ‘yclotexene = 4isoprenyl- ‘methyleyelohexene no“Scu, 595 Limonene, the (S)-enantiomer of d-imonene, with chemical formula CigHg, has a clear colourless liquid form at room temperature with a piney turpentine-like odor. It has a molecular mass of 136.24 g/mol, melting point of -95.2°C, boiling point of 176°C, density of 0.6411 g/cm’, refractive index of 1.4730 and flash point of 50°C. 1-Limonene is also used as botanical insecticide (hitp://wikipedia org). ‘Oktariani (2005) has investigated that the essential oil of C. nobilis containing 57% Himonene was effective against Aedes aegypti with LC = 1301 ppm. The content of Himonene in the fruits varies; therefore the LC ofthe oil also varies. Further study showed that the non-volatile fraction, which was extracted from the residue after distillation by Soxhlet for 10 hours which gave the highest yield 0.36% (w/w) proved effective as repellent and larvacide on adult mosquito and larvae ‘The investigation of insecticidal compounds in jeruk nipis (C. aurantifolia) has been done by Sulacha (1995) who found limonene as the major insecticidal component in the essential oil and 2-hydroxy,3- methyl benzofuran in the nonvolatile fraction effective against rice bug. In this study, the samples were collected as the waste product from the beverage home industry in Kuningan, West Jawa. To ensure that the active components were present, the oil and the non-volatileextract ate analyzed by gas liquid chromatography - ‘mass spectroscopy (GCMS). The formulation of the insecticide, repellent and larvacide is based on the content of the active compounds. The aim of this study is to produce a natural pesticide, which is ‘growing in popularity due to its low toxicity and its ‘environmentally friendly nature ot 2-hidroxy-3-methylbenzofuran Julacha, 1996) Materials and Methods The chemicals used were: organic solvents such as ethanol, hexane, petroleum ether, ethyl acetate; sodium sulfate anhydrous for drying, Repellent formulation consists of: cetaceum, cera alba, liquid pparafine, and sodium tetra borate, Larvacide formulation consisted of: lactose, ethanol, and tween. ‘The raw material was the waste product from the syrup industry at Kuningan, To make the syrup, the ‘whole citrus fruits are cut into two halves and then put into a machine to squeez the juice of it. 1 consists of the peel and the remnants ofthe inner side ofthe fruit. ‘Hydrodistlation was carried out using “karlsure’ ‘Stahl apparatus. Extraction ofthe non-volatile fraction was carried out using Soxhlet extractor. Rotary evaporator was used for evaporating the extract and the usual laboratory glasses for formulation, Analyses ‘was done using GC-17a ver. 3 Shimadzu GCMS-QP ‘5050 class-5000 Release Shimadzu Series Mass Selective Detector equipped with data. processor Hewlet Packard 810 C. ‘The experiment consist of six steps, namely’ sampling of raw materials, distillation, soxhiet extraction, analysis, formulation and bioassay test Raw Material ‘The raw material was the peel of jeruk nipis (Citrus ‘auranifolia) brought from Kuningan as the waste product of a syrup home industry. The fruit peel was cleaned, and washed and divided into four groups. ‘The first group was the peel only cut into small pieces, the second group was the peel made into pulp, the third was the peel and the inner side of the fruit ‘cut into small pieces and the fourth was the peel and the inner side of the fruit made into pulp. Hydrodisiltation ‘The essential oil was obtained from the fresh peel ‘samples were extracted by hydrodistllation using a “Karlsure’ Stahl apparatus anc by steam dis for 20 hours. The oil was separated and ‘sodium sulfate anhydrous and the weight and volume ‘was measured, Proc of Te hematoma Extraction Soxhlet ‘apparatus of nonvolatile fraction by ‘The residue of the peel from hydrodistllation was dried at 60°C in the oven, and then extracted by ethyl acetate Soxhlet for 10 hours. The extract was ‘evaporated by rotavapor until dry, and the weight was ‘measured Analysis by GCMS All of the samples, the oil andl nonvolatile extracts ‘were analyzed by GCMS with the following settings: capillary column DB 5 MS J&W SCIENTIFIC 122. 5532, (30 m x 0.25 mm), temperature program 60- 320°C at 15 °C /min, detector temperatur 320°C, injection volume 0.2 j1L. The oil was not diluted, and the extract was dissolved in hexane in saturated condition. Forrmulation of insecticide, repellent and tarvacide The formulation of the insecticide was available as Space Sprays or Aerosols. In this experiment, plastic bottle with spray nozzle was used instead of aerosol tube. The sprayer was filled with 95% ethanol containing paraben as stabilizer and 0.2% (WAV) citrus oil ‘The formulation of the repellent was done ia two forms: stick and cream, The stick was made by dissolving 10 grams of the extract in ethanol and then ‘mixing it with 10 grams of a mixture of stearic acid, comiperlan, sodium hydroxide, and propylene glycol ‘ne! molded as a stick. The repellent eream was made bby mixing the extract with cetaceum, cerra alba, liquid paratin, sodium tetra borate and aquadest with a 1:1 proportion (Kurniati, 2005), ‘The larvacide was formulated in two ways. First, as granules, which was made by dissolving the extract in ethanol, mixed with lactose (1:1), dried, and then crushed into granules or powder. Second, as a solution, which was made by dissolving the extract in cilsanol, adding tween to make it soluble in water, Bicoassay test of formulas forthe adult mosquitoes and the larvae. ‘The insecticide formula test_was carried out by speaying it into a compartment filled with mosquitoes Dead mosquitoes were counted as the indicator of effectiveness. ‘The repellent formula was tested by smearing it fon the back of a human hand in a compartment filled with 10 starved mosquitoes for 8 hours. The effectiveness of the formula was measured by observing how many mosquitoes still approached to bite, This test was done twice, ‘The larvacide formula was tested by putting it into the water filed with larvae, and the dead larvae were counted after 24 hours. 596 2/4Aaa on tw scence com °° ScienceDirect ELSEVIER Journal of Chomatngapy A, L163 2007) 2-24 wo corse lc coe Review Supercritical fluid extraction in plant essential and volatile oil analysis ‘Seied Mahdi Pourmortazavi*, Seiedeh Somayyeh Hajimirsadeghi Faculty of teria and Manso turing Technologie Malek Aas Univers ofTechnaons PO. Rac H5785 S454, Tekan, ran Received ebay 207: received im revined form | June MAT: cceped Jane 207 ‘Neale line 17 Fame 2007 Abstract “The use af supercritical uid, especially carbon dioxide, the extraction of plant volatile components has increased during mo last decades due to the expected advantages ofthe supercritical extraction process. Superciical uid extraction (SFE) is 2 rapid. selective and convenient ‘method fr sample preparation pric othe analysis uf compound ia the Vola product of plant matnces. Also, SFE‘ simple, inexpensive fat, lfective and virually solvent-free sample pretreatment technique. This weview provides detailed and wpdoted discussion of the developeents, ‘modes and applications of SEE inthe isolation of exseatial oils from plat matices. SFE is usually performed with pure er modified carhon dioxide, which facstates off-line colletion of extracts snd oa-lise coupling With other aalytical methods such as gas, quid apd superceiical ‘uid chromatography: Inhis review. we showed thal a number af factors influence extraction yields, hese beang slublity af the solute the Mud. difusion though the matrix and collection process. Finally, SFE has Been compared with conventional extraction methods in terms of selectivity. ‘rapidity cleanliness and possibility of manipulating the composition ofthe extract. {© 2007 Elsevier BV, All sights reserved -Keponolis Supensitica id extmction; Eset ilk; Antoun; Canenods;Texpenes; Voile componente Contents Inuexduetion . ce - Se Gneeunie Inosuetion io supercritical uid extracuan Sennen cane Solubalicy and mass-transfor rate of plant oils fn supercritical uid . 7 i Effectof matrix oa supeeciial uid extraction ae = 7 ester Effect of extraction parameters... . ne . ae aa 7 5.1. Bflect of pressure and temperature. - . —- ce 52. Bflectof moditers apes ad xan - - _ - - $3. Bfestof extraction time 05.0.0 . cee i Sa. Effect of flow rate oe . . . <1 7 SS. Sample panicle size and packing density oo - - a - 5.6. Bfflectof water supercritical uid exractiom ooo : = - 3. Daying effect - oo en : om - - iterations: AME, alkyporfrin; AIDS, squire immmunedefiiency symleome: AZA-A, aadinchin A; BHT, btylated bydronyiaiuene; CER, coma ‘exaton at CWO, cedar oY, concertabon athe sla th supetiical phase; CC-SFE,coumercurea supercoil had et acon IAD, hada ‘diction, BPP I snheny-2 pry thysharl 4, eteraa mass tanafercoeffixtent, FID, lame ionization etetoe, ACO, pas chroratograghy las tometery: ‘HD. ydotatoa: HS-SPME, headspace sla phase micoettaction: HE hyperfrn Mb drcarbon menatrpene HS, hydocuton sesquiterpene: LC-COs. liquideafvon code; feu, maw extractat constantetaction at, MIS, mse ypecuoencry: MERC. mili letokintc cromaogragy: MWHD. microwsne ‘stint hydratillation; ODS, ectadeyluiica: OM onypemed monouspen: OS, onygenued sexuierpene: solvent fom ite BPLC, revered phe ad bromatography; Re, Reyialds uit, SDE, simultanesus dtllnon tration SFC. apenttical fie ehromabyraphy. SFE. spetcnteal uid extaton: SSW, St Jbu's War (pericam peefovaran | SI, solvent x-eed tao, SPE, sol-pase extraction: SHS, tic Deadspace: ce. ume of the CER pei: ‘salblity:Yena cl concentration at the cu: Vierora, ft amount of sos elected fer he eis memoring same * Comesgening autor Te +98 2122952285; a: 498 2122916878, Email odes: poureetazaye6 yahoo con (3. M, Powmoeta as (021.86 785 ace front mater © 207 Ever BV. All sights oer 410.1016 ons 20070621SM Pourmoriacint 8S Majmirradegh £1. Chromasoge A S148 (3007) 3-24 a 6. Collection of extracted analyte. a 6.1. Salventeolleetion........ - 62. Solid-phase collection ...2..002..000 63. Callection in empty vessels : 6A. Nove collection methads 63, Online coupling of supercritical Mud extraction With chromatographic techniques .....00 0.0 ses ceeso000 7. Exteuction of oxygenated compounds frm plant materials by supercritical uid extraction . 8. Extraction of terpenes and sesquiterpene by supercritical usd extraction - - son of supercritical uid extraction to conventional methods. ae co 5. Con 10. Concise, : - References - cone 1, Introduction [Essceial oils eepresent a small faction of a plant's compo sition bat confer the characteristic for which aromatic plants are used in the pharmaceutical, food and fragrance industries Essential oils have a complex composition, containing from a few dozen to several hundred constituents, especially hydeocar- bons (terpenes and sesquterpens) andl oxygenated compounds (alcohols, aldehydes, ketones, acids. phenols, oxides. lactones aceialse. cers and esters). Both hydrocarbons and oxygenated compounds are responsible for the characteristic odors and flavoes. The proportion of individual compounds in the ol com position is different from trace levels to over 90% (3-Kimanene ‘norange oi) The aroma’sil isthe result ofthe combination of the aromas ofall components Trace components a important, since they give the oil a characteristic and astural odor. Thus js important that the natural proportion of the components ix ‘maintained during extraction ofthe essential ols from plants by any procedure [1]. ‘Steam distilation has traditionally been applied for essential oils recovery from plant materials. One of the disadvantages of the hydodistilation methods is that essential cils undergo chemical alteration and the heat-senstive compounds can easily be destroyed. Therefore. the quality of the essential cil exaracts js extremely impaired (2. “The extraction of essential oil componcnts using solvents af high pressure, or superoitical uid. has received much atten- tion in the past several years, especially in food, pharmaceutical and cosmetic industries, because it presents an alternative to conventional processes sich as organic solvent extraction and seam distillation [3] “The increasing use of vegetable extracts by the food, cos- metic, and pharmaceutical industries can make the extraction of essential oils using supercritical carbon dioxide an attractive technology compared to conventional pracesses with respect to the product quality 4-6]. The knowledge of the mass-ransfer mechanism, the Kinctcs parameters and the thermodynamics restrictions of the extraction conducted in a bed of vegetable material san be used to economically evaluate the extrac- tion process. Tis requires information an the thermodynamic restrictions ofthe system vegetable materia/COs. On the other hand, the understanding of the various process variahles and how ibey can be connected to a theorstical mode! to describe the extraction kinetics are also desiable ‘variety of applications such as essential oil extraction (3). metal ‘cation extraction [7.8]. polymer synthesis[9] and particle nucle- sation [10,11]. In practice, more than 90% of all analytical ‘supercritical fluid extraction (SFE) is performed with carbon dioxide (CO;) for several practical reasons. Apart from having relatively low critical pressure (74'bar) and temperature (32°C), ‘CO, is relatively non-toxic, non-flammable, available in high ‘purity at relatively low cost, and is easily removed from the ‘extract. In the supercritical stale, CO: has a polarity compara- Ibe ts liquid pentane and is, therefore, best suited for lipophilic ‘compounds. The main drawback of COs is its lack of polarity ‘forthe extraction of polar analytes (12). In the 1990s, some reports were published about the choice of ‘N;Oasextraction fluid for analytical SFE [13,14]. This uid was ‘considered better suited for polar compounds because of its per- ‘manent dipole moment. One of the applications where N2O has ‘shown significant improvements when compared te CO> is for le the extraction of polychlorinated dibenzadioxins from ‘ly ash [13]. Unfortunately. this fluid has been shown to cause ‘violent explosions when used for samples having high organic ‘content and should, therefore, be used only when absolutely necessary [13,14]. ‘Other more exotic supercritical fluids which have been used ‘for environmental SFE are SF and freons. SF is a non-polar ‘molecule (although easy polarizable) andas a supercritical uid. ‘it has been shown to selectively exact aliphatic hydrocarbons ‘upto around C-24 from a mixture containing both aliphatic and aromatic hydrocarbons. Freons, especially CHCIF; (Freon-22), ‘has on several occasions been shown to increase the extraction sfliciency compared to conducting extractions with CO» [15] ‘Although supercritical HO’ has often been used for the destruction of hazardous organics, the high temperature and pressure needed (F'> 374°C and P' 221 bar) together with the ‘corrosive nature of HQ at these conditions, has limited the pos- ‘sible practical applications in plant oil analysis [16]. HzO at ‘subcritical conditions has, however, been shown to be an effc- ‘ive fluid for the extraction of several classes of exsential oil. In 2000, Gamiz-Garcia et a. [17] tested a static—dynamic subcrit- ‘cal water extraction for the isolation of fennel oil, Their results ‘showed that, subsritical water eatraction is an efficient method fn terms of quantitative composition of the extract.KARAKTERISASI SENYAWA KIMIA DAN UJIAKTIVITAS ANTIBAKTERI MINYAK ATSIRI BUNGA KECOMBRANG (Etlingera elatior) YANG DIISOLASI DENGAN DESTILASI STAHL Mawar Indah Br Perangin Angin Sekolah Tinggé Penyuluhan Pertanian Medan Ji Binjai Km 10 Tromal Pas 18 Medan 20002 ‘mawarperanginangénS0@yahoo.com ABSTRACT Characterization of chemical compound and antibacterial activity of essential oil of Etlingera elatior flower that obiained from district of Nang Belawan sub district of Simpang Empat in Kara Regency have studied. This research ‘aims to analyses characterization of chemical compounds of antibacterial essential oils and to measure the antibacterial activity of essential oils obtained from flowers kecombrang with some hacteria. Cork Borer methads were used to Determine the activity of antibacterial while analyses of chemical component measured by Infra-Red Spectrophotometer (FT-IR) and Gas Chromatography-Mass. Spectrophotometer (GC-MS). Characterization of Chemical compound were obtained by chromatogram indicated that there were 3 compounds antibacterial essential! oils whick are combination of monoterpene and sesquiterpene. they are a-Pinene (17, 60%), f-Pinene (0,22%), and Campior (0.2974). Volatile oif have antimicrobial activity specially for Salmonella typhi with the highest zona of inhibition 1.9 mum and index antimicrobial 2,6 and also for Shigella sp with inhibition 1,8 mm and index antimicrobial 26 Keywords: Flower of Euingera ¢., essential ail, GC-MS, Antibacterial kaalaa. (Darwis dkk, 1991). Klasifikasi_ilmiah tanaman kecombrang adalah sebagai berikut PENDAHULUAN vans a Kerajaan ‘ndonesia mempunyai banyak jenis tanaman —— Kerainan Plantae yang mengandung minvak atsiri seperti minvak ivi Magnoliophyta sereh, minyak daun cengkeh, minyak kenanga, Kelas Liliapsida ‘minyak akar wangi, minyak kayu cendana, minyak —— Qrdo Zingiberales nnilam dan sebagainya, sehingea Indonesia Famili Zingiberacene ‘menduduki peringkat tertinggi dalam perdagangan Genus Ealingera untuk sejumlah minyak atsiri, Kebanyakan minyak Species Etlingera elatior asin tersebut diekspor atau dijual ke Fuar negeri ke Negara Jepang, Amerika Serikat, Inggris dan Honjefhecombrang —berwarna _kemerahan Eropa (Sastrohamidjojo, 2004), seperti jenis tanaman hias pisang-pisangan atau Tumbuhan kecombrang (Elingera elatior) merupakan tumbuhan yang tersebar cukup las di Indonesia. Kecombrang bagi sebagian orang mungkin kurang dikenal. Tanaman ini mirip bunga bias dan beraroma harum segar. Saat berbentuk bunga, warnanya makin cantik dan aromanya makin tajam. Hampir seluruh bagian dari tumbuhan ini bisa dimanfaatkan. Di Jawa tumbuhan ini dinamakan Kecombrang, Sunda Honje, Sumatera Utara: cekala, kincuang dan sambuang (Minangkabau), Gayo Aceh : kala, tere, Sulawesi : Arimengo, Sitlavo, katimbang serta siantan (Malaya). Orang Thai menyebutnya mirip sekali dengan tanaman lengkuas / laos. Jika batang sudah tua, bentuk tanamannya mirip jahe, dengan tinggi mencapai 5 m, Batang -batang semu bulat gilig, membesar di pangkalnya; tunmbub tegak dan banyak, berdekat-dekatan, membentuk rumpun jarang, keluar dari rimpang yang menjalar di bawab tanh, Rimpangnya tebal, berwama krem, kemerah-jambuan ketika masil muda. Daun 15-30 helai tersusun dalam dua baris, berseling, di batang semu; helaian daun jorong lonjong, 20-90 em» 10 20 em, dengan pangkal membulat atau bentuk jantung, tepi bergelombang, dan ujung meruncing pendek, gundul namun dengan bintik-bintike halus28 dan rapat, hijau mengkilap, sering dengan sisi bawah yang keunguan ketika muda. ‘Di Tanah Karo, buah honje muda disebut asam cekala. Kuneup bunga serta "polong” nya menjadi bagian pokok dari sayur asam Karo; juga menjadi peredam bau amis sewaktu memasak ikan. Masakan batak populer, arsik ikan. mas, juga menggunakan asam eekala ini, Di Pelabuhan Ratu, buah dan bagian dalam pucuk honje sering digunakan sebagai campuran sambal_ untuk menikmati ikan faut bakar Kegunaan minyak atsiri sangat luas dan spesifik, khususnya dalam berbagai_bidang industri, antara_ lain dalam industri kosmetik (sabun, pasta gigi, sampo, losion); dalam industri makanan sebagai bahan penyedap atau penambah cita rasa; dalam industri parfum sebagai pewangi; dalam industri farmasi atau obat-obatan sebagai antinyeri, antiinfeksi, pembunuh bakteri dalam industri bahan pengawet; bahkan digunakan pula sebagai insektisida, oleh karena itu tidak heran jika minyak atsiri ‘banyak diburu berbagai negara (Lutony & Rahmayati,1994). Brum dkk., (1997) juga telah melaporkan bahwa minyak atsiri banyak digunakan sebagai sumber obat-obatan seperti yang dilaporkan dalam hasil uji aktivitas dari minyak atsiri dalam tumbuhan Cochlospermum regium Pilg sebagai antibakteri, Santos dik, (1997) melaporkan minyak atsiri dari tumbuhan Paidium guaianense dapat digunakan sebagai anti radang dan altivitasnya sebagai analgesik Senyawa anti bakteri-merupakan senyawa yang mempunyai kemampuan —menghambat ertumbulian mikroorganisme, senyawa ini dapat berasal dari bagian tanaman seperti bunga, bili, buah, rimpang, batang, dan umbi, Sebagian besar senyawa anti mikroba yang. berasal dari tanaman diketahui merupakan metabolit sekunder terutama dari golongan fenolitik dan terpena dalam minyak atsiri, Beberapa senyawa yang bersifat anti mikroba dari tanaman diantaranya adalah fitoeleksin, asam organik, minyak atsir, fenolitik dan beberapa kelompok —pigmen tanaman. (Naufalin, 2005), Agrica Ekstensia, Vol. 9 No. I Juni 2018: 27-33 Gambar 2. Kuncup bungaKecombrang Chan, dk, (2007) melaporkan bahwa daun dari kecombrang mengandung kadar fenolik yang tinggi dan asam askorbat, juga dapat digunakan sebagai antioksidan dan menghambat aktivitas tirosin. Wong dkk., (1993) meneliti minyak atsii dengan metode destilasi uap terisolasi dari tunas bunga muda kecombrang, Dari hasil penelitian diperoleh bahwa Komponen utama minyak atsii terdiri dari senyawa aldchid alifatik dan alkohol dengan dodecanol dan dadecanal sehagsi dua komponen yang paling banyak. Jaafar, dkk., (2007) juga telah meneliti minyak atsiri yang terkandung, pada daun kecombrang yaitu 8 pinene (19.7%), ‘kariopilen (15,36%) dan sebagai senyawa utama 8- farmesen (27.50%) sedangkan minyak atsiri pada hbatang sebagian besar didominasi oleh 1,1- dodecanediol diasetat (34,26%) dan dodecan (26,99%). Minyak atsiri dari bunga dan rimpang mengandung seayawa utama_yaitu 1,1 - diasetat dodecanediol masing-masing 24,38% dan 40,37% dan siklododecan masing —masing 47,28% dan 34.45%. Berdasarkan uraian yang telah dikemukakan datas membuat penulis. tertarik untuk meneliti salah satu fanaman penghasil minyak atsiri yang bara yaitu tanaman kecombrang dengan mengisolasi dan karakterisasisifatantibakteri ‘minyak atsiri dari bunganya dengan menggunakan destilasi Stahl, serta menguji aktivitas antibakteri pada beberapa bakteri gram positif dan gram negatif, METODOLOGI Penelitian dilakukan di Laboratorium Kimia Bahan Alam Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Sumatera UtaraIranian Food Science and Technology Research Jeurnal Vol. 11, No.3, Agu. Sep. 2015, p. 236-246 © oleh elt ele pple oleting at tei _Teree oat thoy ake lose Ohmic-assisted hydrodistillation of essential oils from Mentha piperita: a comparison with conventional hydrodistillation Mohsen Gavahian', Reza Farhoosh”, Asgar Farabnaky*, Katayoun Javidnia‘, Fakbri Shahi Received: 2014.03.01 Accepted: 20140721 “Abstract ‘Ohmic-assisted hydrodistillation (QAHD) is an advanced hydrodistillation (HD) technique utilizing ohmic heating process and could be considered as a novel method for the extraction of essen oils, Long extraction time is one of the major problems with traditional methods. OAHD of essential oils from the aerial parts of peppermint was studied and the results were compared with those af the conventi ial HD. The results showed that OAHD methed had the extraction time of 19.71 min while this value was about 55.88 min for HD. Scanning electron micrographs of mint leaves showed a sudden eruption of essential oil glands for OAHD samples. GC- MS analysis did nat indicate any noticeable changes in the compounds of the essential oils obtained by OAHD in comparison with HD. The results introduced OAHD as a green technology. Keywords: Essential oils, GC-MS, Hydrodistllaion, Ohmic-assisted hydrodistllation, Mentha pipevita, Introduction Mentha piperita L. (peppermint), a perennial aromatic/medicinal plant belonging. ‘to the family Lamiaceac, is a hybrid of M. spicata L. (spearmint) and M. aguatic (The Wealth of India, 1962).This plant was cultivated by the ancient Egyptians and documented in the Icelandic pharmacopoeia of ‘the thirteenth century. It is widely grown in ‘temperate areas of the world, particularly in Europe, North America and North Africa but ‘nowadays cultivated in other regions of the world including Middle East (Briggs, 1993; The Wealth of India, 1962), ‘The essential oil of Peppermint is commercially used in food, beverage, pharmaceutical, cosmetics, health and tobacco industries (Carmines, 2002). Tobacco industry consumes about 40% of the total mint essential 1, 2 and 5, PRD Student and Professors, Department of Food Science and Technology, Faculty of Agriculture, Ferdowsi University of Mashhad, 3, Associate Professor, Department of Food Science and Technology, School of Agriculture, Shiraz University, Shiraz Iran 4, Professor, Medicinal & Natural Products Chemistry Research Center, Shiraz University of Medical Sciences Shiraz, Iran (* Corresponding author Email: rfarhoosh@um ac ir) oil followed by pharmaceutical and confectionary industries. Usually the major components of peppermint oil include menthol, menthone and menthofiuran. Menthol, the main substance that gives the mints their characteristic aromas and flavors, is used as a raw material in toothpaste, toothpowder, chewing tobacco, confectionary, mouth fiesheners, analgesic balms, cough drops, perfumes, chewing gums, candies and tobacco industry (The Wealth of India, 1962) ‘The peppermint oil is reported to have anti- oxidant properties (Ribeiro et al. 2002), antibacterial activity (Singh et al., 2011) and is one of the most important constituents of some over-the-counter remedies in Europe for irritable bowel syndrome (Pittler and Ernst, 1998) Essential oils can be isolated using a number of methods, eg. hydrodistillation (HD), steam distillation (SD) and organic solvent extraction (Presti_ et al., 2005). Traditional methods for the extraction of essential oils from medicinal plantsherbs are known to be SD and HD. However, these methods suffer fiom some disadvantages including losses of volatile compounds and Jong extraction times and are known to be energy intensive methods, Furthermore,‘Obmic-asssted hydrodistlation of essential oils from... 237 elevated temperatures can cause partial or full degradation of natural constituents especially monoterpenes which are vulnerable to structural changes under steam distillation conditions. Conventional solvent extraction method is likely to involve losses of more volatile compounds during removal of the solvent (Presti ef af., 2005). Ohmic heating (also referred to as Joule heating, electroheating and electro conductive heating) is defined as a process wherein an electrical current (usually alternating) is passed through materials and can be used to generate heat within the product (Goullieux & Pain, 2005). The heating occurs in the form of internal energy transformation (from electric to thermal which is due to Joule effect) within the material (Sastry, 2005). Ohmic processing enables to heat muterials at extremely rapid rates (generally, from a few seconds to a few minutes) (Sastry, 2005). Electrical conductivity is the main parameter determining the heating rate of an ohmic heating treatment. Ohmic heating only works in water continuous systems (due to necessity ‘of ionic mobility) and materials with low electrical conductivity like oil droplets in mixed systems (ie, aromatic plant leave) can not be heated directly by ohmic process but oil droplets will be heated almost rapidly thru heat transfer from the direct neighborhood which however is very nearby due to the small dimensions (Goullicux & Pain, 2005) Distilled water is an excellent electrical insulator so in many studies on ohmic technology, salted water is used as the liquid phase for ohmic treatments (Goullieux & Pain, 2005). In the nineteenth century, several patents were filled for the use of direct resistance heating for the sterilization of static liquid foods (Jones, 1897; Roberts, 1900). In 1993, with the Food and Drug Administration (EDA) approval, process of stable low acid foods at ambient temperature became legal. Since then, ohmic heating has been used ‘commercially in Japan, the USA and Europe. Among novel extraction methods of essential ils, utilizing microwaves for the extraction has been widely studied. Different extraction methods including HD, microwave- assisted hydrodistillation — (MAHD), simultaneous distillation-solvent extraction (SDE) and supercritical fluid extraction (SFE) were reported by Stashenko et al. in 2004 for the extraction of essential oils from Colombian Xylopia aromatica (Lamarck) (Stashenko et al, 2004). MAHD was compared with HD in the extraction of essential oils ftom Thymus vulgaris L. (Golmakani & Rezaei, 2008). ‘Ohmic heating technology presents a large number of actual and potential future applications, including its use in blanching, evaporation, dehydration, fermentation, extraction (USA-FDA, 2000), sterilization, pasteurization and heating of foods to serving temperature in the military field or long- duration space missions (Sastry er al., 2009). Ohmic extraction is one way of using ohmic heating in an extraction process. Ohmic heating as an alternative extraction technique of medicinal plants was first reported by Sensoy and Sastry (2004). Since then, numerous studies have sought the applicability of this new approach for the extraction of materials (Lakkakula et al, 2004). Gavahian et al, (2011) used a combination of ohmic heating and distillation for separation of essentials oils from Zataria multiflora Boiss. (Shirazi thyme) and found significant reductions in extraction time and consumed energy for ohmic-assisted hydrodistillation (OAHD) compare to-——traditional hydrodistillation method. This research team also reported similar results on OAHD for Thymus vulgaris and Myrtus communis (Gavabian ef af, 2012-2013). Nonetheless, due to the lack of enough information it seems that more research is needed to fully explore advantages and disadvantages of OAHD for extraction of essential oils fiom other medicinal plants. Despite many studies reported on the different methods of extraction of essential oils fiom M. piperita, there is no report on the extraction of essential oils from this medicinal plant using OAHD. Therefore, the aim of this work was to use OAHD technique for the extraction of essential oils ftom dried