CHARACTERIZATION AND BIOLOGICAL ACTIVITY OF ESSENTIAL OILS FROM ETLINGERA SPP. Aisyaidil Binti Hanri Bachelor of Science with Honours (Resource Chemistry) 2006 Pluat 1Gl4IUt Maklumat ,ualkmik UNlVERSm MALAYSiA SAaAWJ\K CHARACTERIZATION AND BIOLOGICAL ACTIVITY OF ESSENTIAL OILS FROM ETLINGERA SPP. AISYAIDIL B1NTI HANRI Thi s report is submitted in partial fulfi llment of the requirements for the degree ofBaehelor of Science wi th Honors in Resource Chemistry Faculty of Resouree Science and Teehnology UNIVERSITI MALAYSIA SARAWAK 2006 ilwtKhldlUt Maklumat AXatitmUc UNJVUSm MALAYSIA SAItAWAJ( CONTENTS Contents Page Declaration II Acknowledgement III Abstract IV Abstrak V Li st of Figures VI Li st o f Tables V III Chapter 1 Introduction 1.1 Introduction 1.2 Objectives 4 Chapter 2 Litel'a ture Review 2.1 Essential Oils 5 2.2 Extraction of Essential Oil s 6 2. 3 Zingiberaceae 8 Chapter 3 Materials and Methods 3.1 Plant Materials II 3.2 Extraction of Essential Oil 12 3.3 Instrumental Analysis 3.3. 1 Gas Chromatography- Fl ame Ion Detector (G C/FID ) 12 3.4 Data Analysis 3.4.1 Percentage o f Essential Oil 13 3.4.2 Kovats Indices 14 3.4.3 Semi-quantitative Analysis 15 3.5 Cluster Analysis 15 3.6 Biological Acti vity Study 16 Chapter 4 Results and Discussion 4.1 Percentage of Essential Oi l 17 4.2 Kovat Index of Compounds in Essential Oil s on DB-5 Co lumn 17 4.3 Chemical Composition of Essential Oil 20 4.3. 1 Ellingera elalior 20 4.3.2 Ellingera lilloralis 27 4. 3.3 Ellingera punicea 3.1 4.3 .4 Etlingera lali/abris 40 4.3.5 Etlingera species I 46 4.3.6 Ellingera species 2 53 4.3.7 Comparison of Chemical Composition in leaf oils of six 58 Ellingera species 4. 3.8 Comparison of Chemical Compositi on in rhizome oil s of 58 six Ellingera species 4.3.9 Comparison with other Species and Genus in Fami ly 72 Zi ngi beraceae 4.4 Statistical Analysis of Essential Oi ls fro m Etlingera spp. 74 4.5 Toxicity of Esenti al Oi ls from Ellinf!,era to Brine Shrimp 79 Chapter 5 Conclusion 80 References 81 DECLARAnON No potion of the work refe rred to in this di ssertation has been submitted ill support of all • application for another degree of qualification of thi s or any other university or institution of hi gher learn in g. Aisyaidil binti Hanri Programme of Re:;o urce Chemistry Faculty of Sci ence and Technology Uni versiti Malaysia Sarawak " ACKNOWLEDGEMENT Fi rst, thank to Allah for hi s bless ing I can successfully completing this researc h. I wou ld like to express my spec ial thanks to my supervisor, Assoc. Pro f. Dr. Zaini Bin Assim for giving me supports and guidance in doing this research project. A lso my special thanks to my co-supervisor. Assoc. Prof. Dr Fasihudin B. Ahmad for guiding and supporting me in thi s research. To colleagues at Faculty of Resource Science and Techno logy UNIMAS and all laboratory assistances, in particular En Rajuna Tahir, th anks you for a ll the co-operations given to me during the progress of this project. My appreciation to a ll my lectures and my course mates for he lping and su pporting me while doing this research. iii ABSTRACT Essent ial o il s tram six species of Etlingera were studied fo r their chemical composition and their biological activity on brine shrim p, A. salina. Essentia l oil s were extracted by hydrodist ill ati on and subsequently analyzed usin g gas chromatographylflame ioni zation detector (G C/FlD). The percentage of essentia l oil obta ined from th e six species of Etlingera ranged from 0% to 2.15%. with the hi ghest percentage was obtained from th e leaves of Erlingera Species 2. Eth yl-(E,Z)-decanoate (79.53%), a -gurj unene (1 7.25%), geosm In ( 17. 25%) and tetradecyl aldehyde (8.5 7%) were the major constituent of E. clatior. a -pi nene and ~ - pin e n e were detected at high abundance in E. iil/oralis with 20.67% of a-pi nene in leaf and 20.24% of ~-p i nene in rhizome. E. punicea was rich in octenol (73 .27%), whil e E Lati/abris contain hi gh abundance of octad ienone (6 6. 55%), methylethylpyrazin e (29.30%) and a-pinene (25.91%). Octanol (3 1.02%) and ~ - p in e n e (3 1. 02 %) were th e maj or constituents fo und in Ellingera species I. Etlingera species 2 has a -pinene (25 .3 9%). methyl laurate ( 16.29%) and phenol ( 15.69%) as the major components. Camphene and (E)-a ­ bergamotene were foun d in leaves of ali Etlingera spp. anal yzed. a -ihu.i ene and 2­ ethylpyri dine were found present in rhizome of a ll Etlingera spp. an alyzed. Leaves of E. elatior and Etlingera species I as well as leaves of E. !illoralis and E. lorilabr;" shows c lose re lat ionship. There was also close relationshi p between rhi zomes of ErlinRcra species 2 and E. lati/abri.\' as well as close relationship beiween E. /i f/oratis and EtLing era spccies I . Toxic it y tests on brine shri mp, A. salina shows o nl y leaf o il from Et lingero species I ( 13.59 fJ,g/Il1L ). fl ower oils from E. punicea ( 13.59 fJ,g/mL), leaf o il fro m E. elatior (56.67 fJ, g/mL) and tl ower o il from E. elatior ( I 00 fJ,g/ mL) have toxicity to brine shrim p. Key words: Ellingera, gas chromatographylflame ionization detector. essential o il s, hydrodistillali on, brine shrimp. IV ABSTRAK KOll1posisi ll1inyak pati daripada enall1 spesies Etlingera telah dikaji kOll1posi si kill1ia dan aktiviti biologinya terhadap anak udang, Arlemla salina. Minyak pati telah diekstrak dengan ll1enggunakan kaedah penyulingan hidro dan seteru snya dianalisis ll1 enggunakan kroll1atograti gas/pengesa n ion nyalaan. Peratusan ll1in ya k pati yang diperolehi daripada enall1 spesies Etlingera adalah dalall1 julat 0% hingga 2. 15%, dengan peratusan tertinggi diperolehi daripada daun Etlingera spesies 2. Etil -(E,Z)-dekanoat (79.53%), a-gurjunen (17.25%), geosll1in (17.25%) dan tetradesil aldehid (8.57%) ll1erupakan kOll1pon en utall1 a dalall1 E. elallor. a-pinena and ~-pinena hadir dalam kelii11pahan yang tinggi dalam t:~ lilloralis dengan 20.67% a-pinena telah dijumpai pad a daun dan 20.24% ~-pinena tel ah dijull1pai paad a rhizom. E. punicea ka ya dengan oeten ol (73 .27%), ll1anakala E. latitabris mempunya i kelimpahan oktadienon (66.55%), metiletilpirazina (29.30%) dan a-pillena (25.9 1%). Oklanol (3 1.02%) dan p-pinena (3 1.02 %) merupakan kOll1ponen ulall1a yang dijumpai dalam Ellingera spes ies I . Ellingera spes ies 2 mempun ya i a-pinena (25.39%), metil laurat (16.29%) and feno l (15.69%) sebagai komponen utama. Kamtena and (E)-a­ bergall10ten dijumpai had ir di dalall1 daun dalam setiap E/lingera sp p. yang dianalisis. a­ thujen dan 2-etilpiridina hadir di dalam rhizoll1 dal am se tiap Etlingera spp. yang dian alisis. Daun E. elalior dan Ellingera species 1 dan juga daun E. Iil/orali.l· dan E. 1((lilaiJris ll1enunjukkan hubun gan yang rapat antara satu sama lain. Selain itu , terdapatjuga hubungan yang rapat antara rhizom Ellingera spes ies 2 dan E. ialilabri.\· disamping rhizom E. iiuorolis and Ellingera species I . Ujian ketoksikan terhadap anak udang A. salina l11 enunjukkan han ya ll1in yak pati daripada daun Etiingem species I ( 13.59 Ilg/mL). bun ga E. punicea (13.59 flg/Il1L), daun E. elatlor (56.67 Ilg/mL) dan bunga E. ela/ior ( 100 Ilg/mL) menllnjllkkan ketoksikan terhadap anak udang. Kala kunci: Ellingera, kromatografi gas/pengesan ion nyalaa n, minyak pati, penyulingan hidro, anak udang. v LIST OF FIGURES Figures Pa gc Figure 4.1 Gas chromatogram traced by GC/FID ana lysis of n-alkane 18 sta ndard mixture. Figure 4.2 Gas chromatogram obtained from GC/FID ana ly sis for leaf oil of 21 E. e/olior Figure 4.3 Gas chromatogram obtained from GC/F ID ana ly sis for nower oil 22 of E. elalior Fi gu re 4.4 : Gas chromatogram obtained from GC/FI D analys is for shoot oil 22 of E. elalior Fi gure 4.5 Gas chromatogram obtained from GC/FID analys is fo r rhi zome 23 oiI of E. elaLior Fi gure 4.6 Gas chromatogram obtained by GC/FID analys is for leaf oil of E. 28 lilloralis Figure 4.7 Gas chromatogram obtai ned by GC/F ID analysis for rhizome oi l 28 of E. lilloralis Figure 4.8 Gas chromatogram obtained by GC/FID analysis for leaf oi I of E 34 punicea Figure 4.9 Gas chromatogram obtained by GC/FID ana lys is for stcm oi l ol­ 34 E. punic-ea Figure 4. 10: Gas chromatogram obtained from GC/ FID analys is nower oil of 35 E. puniceo Fi gure 4. 11 Gas chromatogram obtained by GC/FID analys is for rhi zo me oil 35 of E. pUl1icea Fi gure 4.12: Gas chromatogram obtained from GC/FID ana lysis for leaf oil of 41 E. lalilabris Figure 4.1 3: Gas chromatogram obtained from GC/FID analys is fo r stem oil 41 of E. lalilabris Figure 4.14: Gas chro matogram obtained from GC/FID ana lys is for rhi zome 42 oi I of E.