Persicaria odorata Antibacterial Activity of the ... - ResearchGate
Persicaria odorata Antibacterial Activity of the ... - ResearchGate Persicaria odorata Antibacterial Activity of the ... - ResearchGate
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 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. <strong>Persicaria</strong> <strong>odorata</strong> is a native plant <strong>of</strong> sou<strong>the</strong>astern Asia. The taste <strong>of</strong> <strong>Persicaria</strong> <strong>odorata</strong> isdescribed as being pungent and spicy, and its smell is similar to coriander with hints <strong>of</strong> a lemon scent. Volatilecompound from essential oil <strong>of</strong> <strong>Persicaria</strong> <strong>odorata</strong> contains aldehyde such as decanal (Anonymous, 2010).Fur<strong>the</strong>rmore, <strong>the</strong> plant was extracted by different condition and extraction methods. The difference inorganoleptic pr<strong>of</strong>ile indicates a difference in <strong>the</strong> composition <strong>of</strong> 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 <strong>of</strong> <strong>the</strong> essential oil from fresh and dry leaves <strong>of</strong>Vietnamese Coriander (<strong>Persicaria</strong> <strong>odorata</strong>).Materials and MethodsPlant materials: Leaves <strong>of</strong> <strong>Persicaria</strong> <strong>odorata</strong> were purchased from a local vegetable and fruit market <strong>of</strong> <strong>the</strong>Thungkru District, Bangkok, Thailand. For dry sample, <strong>the</strong> leaves were washed to remove soil, peeled,chopped and dried by oven dryer at 40°C for 24 h to reduce <strong>the</strong> water content.Oil Isolation: 1000 g and 500 g <strong>of</strong> fresh and dried leaves <strong>of</strong> <strong>Persicaria</strong> <strong>odorata</strong> were hydrodistilled inClevenger-type apparatus for 24 h. Essential oil was collected and stored in <strong>the</strong> dark vial at 4°C.Physical properties analysis. Physical properties such as percentage <strong>of</strong> yield extraction, color and refractiveindex were observed. Color <strong>of</strong> essential oil was determined using Hunterlab mini scanEZ Color Reader.Refractive index <strong>of</strong> <strong>the</strong> oil was determined by Hand-held Refractometer.Gas Chromatography-Mass Spectrometry analysis: An aliquot <strong>of</strong> 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 <strong>the</strong>n rate at10 o C/min to 140 o C, and <strong>the</strong>n rate at 5 o C/min to 240 o C.<strong>Antibacterial</strong> activity: <strong>Antibacterial</strong> 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 <strong>of</strong>bacteria 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 <strong>of</strong> <strong>the</strong> essential oil that dissolved in steriledimehtylsulfoxide (DMSO) and put in <strong>the</strong> inoculated plates. The inoculated plates were incubated at 37±2 o C for24h and <strong>the</strong>n <strong>the</strong> inhibition zones were measured in diameter (mm) around <strong>of</strong> <strong>the</strong> 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 <strong>the</strong> highest concentration (50 µl/ml) and <strong>the</strong>n serial two-fold dilutionswere made in a concentration range from 6.25 µl/ml to 50 µl/ml. The least concentration <strong>of</strong> each extractshowing a clear <strong>of</strong> inhibition was taken as <strong>the</strong> 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 <strong>of</strong> <strong>Persicaria</strong> <strong>odorata</strong>which extracted by hydrodistillation methods showed a % yield at 0.274% and 0.12%, respectively (Table 1).Yield <strong>of</strong> essential oil from dry leaves was lower than fresh leaves because some <strong>of</strong> volatile compound from dry
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