Persicaria odorata Antibacterial Activity of the ... - ResearchGate

Agricultural Sci. J. 42(2)(Suppl.): 105-108 (2011) ว. วิทย์. กษ. 42(2)(พิเศษ): 105-108 (2554)องค์ประกอบทางเคมี และประสิทธิภาพการเป็ นสารต้านแบคทีเรียของนํามันหอมระเหยจากใบ Persicaria odorataAntibacterial Activity of the Essential Oil from Persicaria odorata Leavesพราโมโน สาสงโก้1,2 ณัฎฐา เลาหกุลจิตต์2 และ อรพิน เกิดชูชื น 2Sasongko, P. 1,2 , Laohankunjit, N. 2 and Kerdchoechuen, O. 2AbstractThis study conducted to identify physicochemical properties and antibacterial activity of the essentialoils from Vietnamese Coriander (Persicaria odorata). Hydrodistillation was used to extract the essential oil fromthe fresh and dry leaves of Persicaria odorata. Using a Gas Chromatography-Mass Spectrometry (GC-MSanalysis), dodecanal, caryophyllene, alpha-caryophyllene, drimenol and decanal were identified as a majorvolatile compounds. The antibacterial testing was conducted by disc diffusion method. The essential oils fromboth of fresh and dry leaves of Persicaria odorata exhibited antibacterial activity against both of gram positivebacteria such as Staphylococcus aureus and gram negative bacteria such as Escherichia coli. Moreover, thehighest antibacterial activity was shown by essential oil from dry leaves which against S. aureus exhibited aninhibition zone at 26 mm for 100µl/ml concentration.Keywords: Persicaria odorata, essential oil, gas chromatography-mass spectrometry, antibacterial activityบทคัดย่องานวิจัยนี ศึกษาองค์ประกอบทางเคมี และประสิทธิภาพการเป็ นสารต้านแบคทีเรียของนํ ามันหอมระเหยจากผักแพว (Persicaria odorata) อบแห้งและสด ทีสกัดโดยการต้มกลัน ซึงนํ ามันผักแพวมีสารหอมระเหยได้ วิเคราะห์ด้วยเครืองgas chromatography-mass spectrometry (GC-MS) ได้แก่ dodecanal, caryophyllene, alpha-caryophyllene,drimenol และ decanal เมือนํามาทดสอบประสิทธิภาพการเป็ นสารต้านแบคทีเรียด้วยวิธี disc diffusion method พบว่านํ ามันหอมระเหยจากใบผักแพวสดและแห้ง มีความสามารถยับยั งแบคทีเรียแกรมบวก เช่น Staphylococcus aureus และแกรมลบ เช่น Escherichia coli ได้ นอกจากนี นํ ามันหอมระเหยจากผักแพวอบแห้งมีประสิทธิภาพยับยั งการเจริญของเชื อแบคทีเรีย S. aureus ได้สูงสุด เมือใช้ความเข้มข้น 100µl/ml โดยมีค่า inhibition zone เท่ากับ 26 มิลลิเมตรคําสําคัญ: Persicaria odorata นํ ามันหอมระเหย gas chromatography-mass spectrometry ประสิทธิภาพการต้านเชื อแบคทีเรียIntroductionEssential oils have a significant role in the society where they are variously used in medicine,pharmacy, cosmetics, chemical and food-processing industries (Yentema et al., 2007). Essential oils areimportant constituent of some higher plants comprising monoterpenes, sesquiterpenes, arylpropanoids andother derivatives. The antimicrobial properties of essential oils have been recognized for long time and theyhave been scientifically established (Sadashiva et al., 2010). Essential oils are very complex natural mixtureswhich contain about 20–60 components at quite different concentrations. They are characterized by two orthree major components at fairly high concentrations (20–70%) compared to other components present intrace amounts (Bakkali et al., 2008). Moreover, it has been reported that essential oil has biological activitiessuch as an antimicrobial, antioxidant, antifungal and anticancer activity. The simple and complex derivatives ofphenol are main antimicrobial compounds in essential oils from spices (Conner, 1993). Coriandrum sativumessential oil was reported that has an antibacterial activity against either gram positive or gram negativebacteria (Matasyoh et al., 2009). Furthermore, Laohakunjit et al. (2008) concludes that essential oil from1Faculty of Agriculture Technology, University of Brawijaya, Malang 65141, Indonesia2School of Bioresources and Technology, King Mongkut’s University of Technology Thonburi 83 Moo 8 Tha-kam, Bangkhuntien, Bangkok 10150

106 ปี ที 42 ฉบับที 2 (พิเศษ) พฤษภาคม - สิงหาคม 2554 ว. วิทยาศาสตร์เกษตรKaempferia (Boesenbergia pandurata Holtt.) and Bastard cardamom (Amomum xanthioides Wall) has anantibacterial effect towards Escherichia coli, Staphylococcus aureus, Bacillus cereus and Listeriamonocytogenes. Persicaria odorata is a native plant of southeastern Asia. The taste of Persicaria odorata isdescribed as being pungent and spicy, and its smell is similar to coriander with hints of a lemon scent. Volatilecompound from essential oil of Persicaria odorata contains aldehyde such as decanal (Anonymous, 2010).Furthermore, the plant was extracted by different condition and extraction methods. The difference inorganoleptic profile indicates a difference in the composition of oils which were obtained by different conditionand this may also influence on chemical and volatile properties (Hashemi et al., 2008). This study was aimedto identify physicochemical properties and antibacterial activity of the essential oil from fresh and dry leaves ofVietnamese Coriander (Persicaria odorata).Materials and MethodsPlant materials: Leaves of Persicaria odorata were purchased from a local vegetable and fruit market of theThungkru District, Bangkok, Thailand. For dry sample, the leaves were washed to remove soil, peeled,chopped and dried by oven dryer at 40°C for 24 h to reduce the water content.Oil Isolation: 1000 g and 500 g of fresh and dried leaves of Persicaria odorata were hydrodistilled inClevenger-type apparatus for 24 h. Essential oil was collected and stored in the dark vial at 4°C.Physical properties analysis. Physical properties such as percentage of yield extraction, color and refractiveindex were observed. Color of essential oil was determined using Hunterlab mini scanEZ Color Reader.Refractive index of the oil was determined by Hand-held Refractometer.Gas Chromatography-Mass Spectrometry analysis: An aliquot of 1 µl oils (dissolved in ethanol) was adjustedto 100 ppm after that it was directly injected into GC-MS Agilent 7890A which was carried out in GC-MSsystem, fitted with a 30 m x 0.25 mm i.d. x 0.25 µm film thickness, DB-5ms capillary column. The carrier gaswas helium at flow rate 0.57 ml/min. The injector and detector temperatures were 230 o C and 250 o C,respectively. The running methods were splitless mode, pressure: 3 psi, oven temperature: 70 o C then rate at10 o C/min to 140 o C, and then rate at 5 o C/min to 240 o C.Antibacterial activity: Antibacterial activity assays were carried out by disc diffusion method. OvernightStaphylococcus aureus and Escherichia coli bacterial suspension (50 µl) adjusted to contain 1x10 6 CFU/ml ofbacteria by spectrophotometer compared with MacFarland no. 0.5, which OD must be between 0.008-0.1.Bacterial suspension was spread by a sterile glass spreader on Muller-Hinton Agar (MHA) medium. Sterile 6mm filter paper (Whatman No.1) disc was impregnated with 100µl/ml of the essential oil that dissolved in steriledimehtylsulfoxide (DMSO) and put in the inoculated plates. The inoculated plates were incubated at 37±2 o C for24h and then the inhibition zones were measured in diameter (mm) around of the disc. DMSO was used asnegative control. Streptomycin (5mg/ml) was used for positive control. The assays were performed with threereplicates.Minimum Inhibition Concentration (MIC) value: MIC value was carried out in disc diffusion assay. Plant extractsdissolved in DMSO were first diluted to the highest concentration (50 µl/ml) and then serial two-fold dilutionswere made in a concentration range from 6.25 µl/ml to 50 µl/ml. The least concentration of each extractshowing a clear of inhibition was taken as the MIC levels. DMSO was used as negative control andstreptomycin (5mg/ml) was used for positive control.Results and DiscussionThe essential oil was recovered from 1000 g fresh and 500g from dry leaves of Persicaria odoratawhich extracted by hydrodistillation methods showed a % yield at 0.274% and 0.12%, respectively (Table 1).Yield of essential oil from dry leaves was lower than fresh leaves because some of volatile compound from dry

ว. วิทยาศาสตร์เกษตร ปี ที 42 ฉบับที 2 (พิเศษ) พฤษภาคม - สิงหาคม 2554 107leaves has been vaporized in drying process. Color measurement result showed that the color of essential oilfrom fresh and dry leaves in 2000 ppm concentration was light yellow green. Moreover, refractive index valuesof essential oil from fresh and dry leaves are 1.472 and 1.471, respectively, this refractive index value wassimilar to refractive index of some spice and herbs oil which in range of 1.462-1.600 (Singhal et al., 2001).Gas Chromatography-Mass Spectrometry chromatogram results were compared with Wiley275 andNIST library used a % quality greater than 85%. RI value was calculated with a DB-5ms stationary phase usinga series of n-alkanes standard between C 10 and C 20 . There are 26 volatile compounds contained in essential oilfrom fresh and dry leaves of Persicaria odorata (Table 2). The major volatile compounds from both of fresh anddry leaves are eupatoriochromene (21.71% and 20.94%), dodecanal (19.96% and 18.72%), alphacaryophyllene(12.57% and 11.62%), beta-caryophyllene (11.07 % and 11.40%) and decanal (7.32 % and4.47%), respectively. Others compound such as eucalyptol, alpha,-curcumene, eremophilene and drimenolwere also detected in trace amount.Furthermore, as shown in Table 3, the essential oil from fresh and dry leaves of Persicaria odoratapossess strong antibacterial activity against gram positive bacteria: Staphyloccocus aureus and gramnegative bacteria such as Escherichia coli. Essential oil from fresh and dry leaves produced an average zoneof inhibition (ZOI) of 21 and 26 mm against S. aureus and average ZOI against E. coli are 13 and 19 mm,respectively. Essential oil from Persicaria odorata leaves has been known as source of aldehyde and terpenecompound such as dodecanal, decanal, caryophyllene, eremophillene and alpha-curcumene which arepossess antibacterial properties (Sadashiva et.al., 2010). The lowest concentrations that can inhibit the growthof bacteria were shown by MIC values at 6.25 µl/ml for S. aureus and 12.5 µl/ml for E.coli. Although, averageZOI of the E. coli was lower than S. aureus, the MIC value of essential oil against E. coli was higher thanagainst S. aureus because the essential oil has a better activity against gram positive bacteria (S. aureus) thangram negative bacteria (E. coli) and gram negative bacteria was more resistant. A gram negative bacteriaresistance was influenced by the structure of cell wall. Gram negative bacteria have an outer and innermembrane in their cell wall which is more complicate compared with gram positive bacteria cell membrane(Anonymous, 1999).SummaryThe major volatile compounds of essential oil from fresh and dry leaves of Persicaria odorata waseupatorichromene, dodecanal, alpha-caryophyllene, beta-caryophyllene and decanal. Furthermore, this oil hasa strong antibacterial activity against both of gram positive and gram negative bacteria. The MIC value ofessential oil from fresh and dry leaves of Persicaria odorata against S.aureus and E.coli are 6.25 µl/ml and12.5 µl/ml, respectively.AcknowledgementThis work was supported by the Higher Education Research Promotion and National ResearchUniversity Project of Thailand, Office of the Higher Education Commission.Literature citedAnonymous, 2010, Persicaria odorata [online], available: http://en.wikipedia.org/wiki/Vietnamese_coriander[2011, 11 th February].Anonymous. 1999, Physiology & Biochemistry Of Microorganisms, [online], available: http://www.micro.siu.edumicr425/425Notes/01-Introduction.html. SIUC / College of Science / Microbiology. [2010, November 28 th ].Bakkali, F., Averbeck, S., Averbeck, D. and Idaomar, M., 2008, Biological Effects of Essential Oils – A review,Journal of Food and Chemical Toxicology, 46(2): 446-475.

106 108ปี ที 42 ฉบับที 2 (พิเศษ) พฤษภาคม - สิงหาคม 2554 ว. วิทยาศาสตร์เกษตรConner, D.E., 1993, Naturally Occurring Compounds, pp. 441-468. In P.M. Davidson and A.L. Branen (eds.).Antimicrobials in Foods. 2nd ed. Marcel Dekker, Inc., New York.Hashemi, P., Abolghasemi, M.M., Ghiasvand, A.R., Ahmadi, S., Hassanvand, H. and Yarahmadi, A., 2008, AComparative Study of Hydrodistillation and Hydrodistillation–Solvent Microextraction Methods forIdentification of Volatile Components of Echinophora cinerea, Journal of Chromatographia, 69: 179-182.Matasyoh, J.C., Maiyo, Z.C., Ngure, R.M. and Chepkorir, R., 2008, Chemical Composition and AntimicrobialActivity of The Essential Oil of Coriandrum sativum, Journal of Food Chemistry, 113(2): 526-529.Laohakunjit, N., * Orapin, K., Krittika, N. and Pantip, B, 2008, Essential Oil from Five Zingiberaceae for AntiFood-Borne Bacteria, International Food Research Journal, 15(3): 337-346.Singhal, R.S., Kulkarni, P.R. and Rege, D.V., 2001, Quality Indices for Spice Essential Oils, Chapter 3 pp. 30.InPeter, K.V., Handbook of Herbs and Spices, CRC Press, USA.Sadashiva, C.T., Sharanappa, P., Remashree, A.B., Raghu, A.V., Udayan, P.S. and Balachandran, I., 2010,Chemical Composition and Antimicrobial Activity of the Essential Oil from Bark of Pittosporum dasycaulonMiq., Advances in Biological Research, 4(6): 301-304.Yentema, O., Alioune, O. and Dorosso, S.A., 2007, Chemical Composition and Physical Characteristic of theEssential oil of Cymbopogon schoenanthus (L.) Spreng of Burkina Faso, Journal of Applied Sciences,7(4): 503-506.Table 1 Physical properties of essential oil from fresh and dry leaves of Persicaria odorataSample Yield of extraction L* a* b* c* Hue angle Color appearance Refractive index(%)-dry basisFresh 0.274 70.61 -0.86 2.50 2.64 108.99 Light green 1.472Dry 0.120 69.96 -0.86 2.26 2.42 110.83 Light yellow 1.471Table 2 Constituent of the essential oil from fresh and dry leaves of Persicaria odorataNo. Volatile compounds RI a Composition (%)No. Volatile compounds RI a Composition (%)Fresh Dry Fresh Dry1. eucalyptol 1051 0.31 - 14. alpha,-curcumene 1490 3.16 4.492. undecane 1108 - 0.14 15. eremophillene 1508 6.89 4.203. 1-nonanol 1174 - 0.09 16. 7-epi-alpha-selinene 1537 - 2.594. decanal 1212 7.32 4.47 17. ledol 1550 5.99 -5. decanol 1274 - 3.34 18. nerolidol 1564 - 3.676. undecanal 1311 0.58 0.57 19. globulol 1587 0.95 -7. n-decanoic acid 1358 0.21 - 20. caryophyllene oxide 1601 2.04 5.648 1-nonene 1374 - 2.02 21. cubenol 1640 0.08 -9. beta-elemene 1401 0.64 - 22. eupatoriochromene 1664 21.71 20.9410. dodecanal 1417 19.96 18.72 23. drimenol 1790 4.74 4.3411. beta-caryophyllene 1441 11.07 11.40 24. hexahydro farnesyl acetone 1842 - 0.6012.13.allo-aromadendrenalpha.-caryophyllene145614760.4912.57-11.6225.26.isophytoln-hexadecanoic acid194519590.390.910.170.99a Retention indices calculated based on n-alkane standard (C 10 -C 20 )Table 3 Antibacterial activity and Minimum inhibitory concentration (MIC) value of essential oil from fresh anddry leaves of Persicaria odorataBacterial speciesInhibition zones (mm)*MIC value(µl/ml)Fresh Dry DMSO** Streptomycin (5mg/ml)*** Fresh DryS. aureus 21 26 - 23 6.25 6.25E. coli 13 19 - 20 12.5 12.5*Inhibition zone including the diameter of the paper disc (6 mm); ** Negative control (DMSO); *** Positive control (5mg/ml Streptomycin)