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Oral Presentation CRDC 5 22/07/54 ANTIBACTERIAL ACTIVITY OF ANTIBACTERIAL ACTIVITY OF THE ESSENTIAL OIL FROM THE ESSENTIAL OIL FROM Persicaria odorata LEAVES Persicaria odorata LEAVES Pramono Sasongko Faculty of Agriculture Technology, University of Brawijaya, Indonesia School of Bioresources and Technology, King Mongkut’s University of Technology Thonburi , Bangkok 5 th Tropical & Sub-Tropical Crops Research Symposium Thailand, 2011 Outline Outline Introduction Objective Materials and Methods Results and Discussion Summary 1

Oral Presentation CRDC 5 22/07/54 Introduction Introduction Essential oil Essential oil An essential oil is a concentrate An essential oil is a concentrate, hydrophobic liquid containing volatile aroma compounds from plants � Pharmacy � Medicine � Cosmetics � Cosmetics � Foods processing Industries � Etc. 1 Introduction Introduction Essential Oil’s Production Plant Plant Extraction Extraction Hydro/steam Hydro/steam Solvent Solvent Expression Expression Enfleurage Enfleurage distillation distillation Extraction Extraction Essential oil Essential oil Concretes Concretes Pomades Pomades Essential oil Essential oil Resinoids Resinoids Absolutes Absolutes 2 2

Oral Presentation CRDC 5 22/07/54 Introduction Introduction Essential oil Essential oil Antibacterial Antibacterial Antioxidant Antioxidant Antifungal Antifungal Anticancer Anticancer Antiviral Antiviral Research Type of plant Purpose of use Zheng and Wang (2001) 39 different herbs, including medicinal and culinary herbs Antioxidant Sylvestre, et al (2006) Croton flavens Antioxidant and anticancer Klaric , et al (2006) Klaric´ et al (2006) Thyme ( Thymus vulgaris ) Thyme ( Thymus vulgaris ) Antifungal Antifungal Gende, et al (2008) Cinnamomum zeylanicum antimicrobial Bioactive compounds Bioactive compounds Adiguzel, et al., (2009) Methanol extract of Nepeta cataria Antimicrobial and antioxidant Abdoul ‐ latif, et al (2010) Jasminum sambac Antibacterial and antioxidant Lopez, et al (2011) dried Phyllanthus niruri , dry and fresh Piper betle L. Antimicrobial 3 Introduction Introduction Essential oil Essential oil Aliphatic compounds p p An essential oilis a rich Terpene compounds source of biologically active compounds Benzene compounds Other compounds 4 3

Oral Presentation CRDC 5 22/07/54 Introduction Introduction Essential oil Essential oil 5 Introduction Introduction Persicaria Persicaria odorata odorata � Common name � Common name Common name Common name : Vietnamese coriander : Vietnamese coriander : Vietnamese coriander : Vietnamese coriander � Thai name Thai name : Pak : Pak ‐ Paeow Paeow � Usually use for culinary herbs Usually use for culinary herbs Research result Method of Type of extraction extract extracted with 15 mL Zheng and Zheng, and Crude extract d h has an antioxidant activity d of phosphate buffer Wang (2001) (75 mM, pH 7.0) Polygonum odoratum or Nanasombat persicaria odorata has an Methanol extract Crude extract And Teckchuen antimicrobial activity and (2009) antioxidant activity 6 4

Oral Presentation CRDC 5 22/07/54 Introduction Introduction (Nanasombat and Tekhchuen, 2009) (Zheng and Wang, 1995) Volatile compound from essential oil of Persicaria odorata contains aldehyde such as decanal (Anonymous, 2010). plant was extracted by different condition and extraction methods. The difference in organoleptic profile indicates a difference in the composition of oils which were obtained by different condition and this may also influence chemical and volatile properties (Hashemi et al, 2008). Solvent extraction was show a different composition of volatile compounds from Persicaria odorata leaves extract 7 Introduction Introduction Food borne pathogens S. aureus E. coli Gram positive Gram negative Structure of cell wall bacteria Gra Gram - po positive sitive Gram Gram - negative egative 8 5

Oral Presentation CRDC 5 22/07/54 Outline Outline Introduction Objective Materials and Methods Results and Discussion Summary Objective Objective Identify physico-chemical properties of the essential oil from Persicaria odorata leaves Identify antibacterial activity of the essential oil from Persicaria odorata leaves 9 6

Oral Presentation CRDC 5 22/07/54 Outline Outline Introduction Objective Materials and Methods Results and Discussion Summary Materials Materials Plants Plants Bacteria Bacteria Fresh leaves Gram + of P. odorata ( S.aureus ) D i d l Dried leaves Gram – of P. odorata ( E.coli ) (40 o C for 24h) 10 7

Oral Presentation CRDC 5 22/07/54 Methods Methods Persicaria odorata Extraction Physico ‐ chemical Antibacterial assay Analysis • Washing • Hydrodistillation • Antibacterial activity • Chopping • % yield of extraction • MIC Value • Drying • Refractive Index • Color analysis • GC ‐ MS analysis 11 Outline Outline Introduction Objective Materials and Methods Results Summary 8

Oral Presentation CRDC 5 22/07/54 Results Results Results Results Physical properties Physical properties % yield extraction Color Refractive Index Yield of Color Refractive Sample extraction L* a* b* c* Hue angle appearance index (%)-dry basis 0.274 70.61 -0.86 2.50 2.64 108.99 Light green 1.472 Fresh 0.12 0 12 69 96 69.96 -0.86 0 86 2 26 2.26 2 42 2.42 110.83 110 83 Li ht ll Light yellow 1 471 1.471 D Dry 12 Results Results Results Results Volatile compound composition Volatile compound composition GC ‐ MS analysis GC MS analysis Analyzed using GC ‐ MS Agilent 7890A, equipped with : 1. 30 m x 0.25 mm i.d. x 0.25 µm film thickness, 2. DB ‐ 5ms capillary column. 3. The carrier gas was helium at flow rate 0.57 ml/min, and 1 µml of sample (100 ppm concentration) was directly injected. 4. The injector and detector temperatures were 230 o C and 250 o C respectively. 5. The running methods were splitless mode, pressure: 3 psi, oven temperature: 70 o C then 10 o C/min. to 140 o C, and then 5 o C/min. to 240 o C 13 9

Oral Presentation CRDC 5 22/07/54 Results Results Results Results Volatile compound composition Volatile compound composition GC ‐ MS analysis y � Compared with WILEY257 and NIST library use a % quality match greater than 85% � RI calculation based on n-alkane standard (C 10 -C 20 ) 14 Results Results Results Results Volatile compound composition Volatile compound composition % of % of Composition No. Volatile compound RI a No. Volatile compound RI a Composition Fresh Dry Fresh Dry 1 1. eucalyptol l t l 573 573 0 31 0.31 - 14. 14 alpha,-curcumene l h 1291 1291 3 16 3.16 4 49 4.49 2. undecane 677 - 0.14 15. eremophillene 1416 6.89 4.20 - 3. 1-nonanol 743 - 0.09 16. 7-epi-alpha-selinene 1447 2.59 - 4. decanal 841 7.32 4.47 17. ledol 1459 5.99 - 5. decanol 903 - 3.34 18. nerolidol 1474 3.67 6. undecanal 1018 0.58 0.57 19. globulol 1496 0.95 - 7. n-decanoic acid 1065 0.21 - 20. caryophyllene oxide 1626 2.04 5.64 8 1-Nonene 1080 - 2.02 21. cubenol 1665 0.08 - 9. 9. beta elemene beta-elemene 1203 1203 0.64 0.64 - 22. 22. eupatoriochromene eupatoriochromene 1688 1688 21.71 21.71 20.94 20.94 10. dodecanal 1216 19.96 18.72 23. drimenol 2049 4.74 4.34 11. beta-caryophyllene 1240 11.07 11.40 24. Hexahydro farnesyl acetone 2147 - 0.60 12. allo-aromadendren 1258 0.49 - 25. isophytol 2316 0.39 0.17 13. alpha.-caryophyllene 1276 12.57 11.62 26. n-hexadecanoic acid 2329 0.91 0.99 15 10

Oral Presentation CRDC 5 22/07/54 Results Results Results Results Volatile compound composition Volatile compound composition the major compounds are aldehyde and terpene compounds (Sadhasiva et al, 2010) (Sikkema et al, 1995) Aldehyde and terpene compounds has an antibacterial properties 16 Results Results Results Results Volatile compound composition Volatile compound composition the major compounds are aldehyde and terpene compounds (Mihailovic et al, 2009) Eupatoriochromene, caryophyllene, and caryophyllene oxide was shown an antimicrobial activity 17 11

Oral Presentation CRDC 5 22/07/54 Results Results Results Results Antibacterial assay Antibacterial assay Disc Diffusion Method 50 µl 50 µl Spread by a sterile bacterial glass spreader suspension (1x10 6 CFU/ml) Muller ‐ Hinton Agar (MHA) Measured the incubated at 37 ± 2 o C inhibition zones for 24h (mm) 6mm filter paper impregnated with 100µl/ml of the essential oil from fresh and dry leaves of P. odorata 18 Results Results Results Results Antibacterial assay Antibacterial assay Disc Diffusion Method Inhibition zones (mm)* against Type of Essential oil S. aureus E. coli Fresh leaves 21 13 Dry leaves 26 19 DMSO** - - Streptomycin (5mg/ml)*** 23 20 *Inhibition zone including the diameter of the paper disc (6 mm); ** Negative control (DMSO); *** Positive control (5mg/ml Streptomycin) The essential oil from fresh and dry leaves of The essential oil from fresh and dry leaves of P. P. odorata odorata show show an antibacterial activity against an antibacterial activity against S. S. aureus aureus and and E. coli E. coli . . 19 12