BAB V. KESIMPULAN DAN SARAN
B. Saran
1. Perlu dilakukan pengujian toksisitas ekstrak etanol daun tumbuhan tembelekan menggunakan biakan sel kanker dan sel normal.
2. Perlu dilakukan penelitian lebih lanjut mengenai toksisitas fraksi aktif dari ekstrak etanol daun tumbuhan tembelekan.
3. Perlu dilakukan identifikasi lebih lanjut terhadap jenis flavonoid dan triterpenoid yang diduga menyebabkan ketoksikan ekstrak etanol daun tumbuhan tembelekan.
DAFTAR PUSTAKA
Aida, 1990, Usaha Isolasi dan Identifikasi Komponen Kimia Daun Tembelekan asal Tamalanrea Ujung Pandang, Skripsi, Fakultas MIPA Universitas Hasanudin, Ujung Pandang.
Albert B., Johnson A., Lewis J., Raff M., Roberts K., Walter P., 2002, Molecular Biology of The Cell, fourth edition, 1356-1357, Garland Science, Taylor & Francis Group, AS.
Anderson, J.E., Goets, C.M., and Mc Laughin, J.L., 1991, A Blind Comparison of simple Benzh-top Bioassay and Human Tumor Cell Cytotoxicities as Antitumor Prescreens, Phytochemical Analysis, Volume 2, 107-111.
Anonim, 1985, Cara Pembuatan Simplisia, 2-11,13, Direktorat Jandral Pengawasan Obat dan Makanan, Departemen Kesehatan Republik Indonesia, Jakarta. Anonim, 1986, Sediaan Galenik, 4-5,16-20, Depkes RI, Jakarta.
Anonim, 1995, Materia Medika Indonesia, Jilid IV, xvii, Departemen Kesehatan Republik Indonesia, Jakarta.
Asteria, W.I.S., 2006, Uji Potensi Antibakteri Ekstrak Etanol Daun Tembelekan terhadap Staphylococcus aureus ATCC 25923 dan Escherichia coli ATCC 35218, Skripsi, Fakultas Farmasi USD, Yogyakarta.
Asterina, R., 1994, Pemeriksaan Flavonoid dan Verbakosid Daun Tembelekan,
Skripsi, Fakultas Farmasi ITB, Bandung.
Becker, C.A., Bakhuizen Van den Brink, R.C., 1963, Flora of Java, Vol I, 3-6, 29-34, N.V.P. Noordoff, Groningen, The Netherlands.
Cheng, R.Z., 2003, Senyawa Antikanker, http://www.kompas.com/kompas-cetak/0310/30/inspirasi/657330.htm. Diakses tanggal 1 April 2006.
Dalimartha, S., 1999, Atlas Tumbuhan Obat Indonesia, Jilid I, 154-157, Trubus Agriwidya, Ungaran.
Dalimartha, S., 2003, Ramuan Tradisional Untuk Pengobatan Kanker, 1-4, Penebar Swadaya, Jakarta.
Donatus, I.A., 1990, Toksikologi Pangan, Edisi I, 247-248, PAU Pangan dan Gizi UGM, Yogyakarta.
Evans, WC., Trease, 2002, Pharmacognosy, 15th edition, 113-114, 297-298, 324, W.B. Saunders, New York.
Ganong, W.F., 1995, Buku Ajar Fisiologi Kedokteran, 9-35, Penerbit Buku Kedokteran ECG, Jakarta.
Halang, B, 2004, Toksisitas Air Limbah Detergent terhadap Ikan Mas, Bioscientiase, Vol 1 No1, Januari 2004, 39-49.
Harborne, J.B., 1984, Phitochemical Methods, diterjemahkan oleh Kosasih Padmawinata, 71-76, 123,125, Penerbit ITB, Bandung.
Harborne, J.B., 1986, The Flavonoids, 643-644, Chapman & Hall, USA.
Hembing, W., 2000, Ensiklopedi Milenium Tumbuhan Berkhasiat Obat Indonesia, 159-163, Prestasi Insan Indonesia, Jakarta.
Kaufman, P.B., Cseke L.J., Warbers, Duke, J.A., Brielmann, H.L., 1988, Natural Products from Plants,16, CRC Press, New York.
Kumar, S., Prasad, S., and Singh,R.N., 2005, Resurgence of Spider Mite Tetranychus ludeni Zacher (Acarina: Tetranychidae) Against Acaricides and Botanical Pesticides on Cowpea, Resistant Pest Management Newsletter.
Lee, Yue-Wei; Fang, Qi-Cheng; Wang, Zhen-Guo; Li, De-Hua; Cook, C. E.; 1991,
Pentacyclic triterpenoid compounds as topoisomerase inhibitors or cell differentiation inducers, http://www.freepatentsonline.com/5064823.html. Diakses 25 Januari 2007.
Loomis, T.A., 1978, Essential of Toxicology, Edisi III, diterjemahkan oleh Imono Argo Donatus, 228-233, IKIP Semarang, Semarang.
Mabry, T.J., Markham, K.R., and Thomas, M.B., 1970, The Systematic Identification of Flavonoids, 13, Springer Verlag, Berlin.
Markham, K.R., 1988, Techniques of Flavonoid Identification, diterjemahkan oleh Kosasih Padmawinata, 15-17, Penerbit ITB, Bandung.
Meyer, B.N., Ferrigni, N.R., Putnam, J.E., Jacobsen, L.B., Nichols, D.E., and McLaughlin, J.L., 1982, Brine Shrimp: A Convenient General Bioassay for Active Plant Constituents, Planta Medica, Vol. 45,31-34.
Michael P., 2007, Biochemical Pharmacology, University of Waterloo.
Middleton, E., Kandaswami, C., and Theoharides, T.C., 2000, Pharmacological Review, vol 45, 673-751, Tufts University School of Medicine, Boston.
Mills, S., Bone, K., 2000, Principles and Practice of Phytotherapy, 157-160, Churchill Livingstone, New York.
Mudjiman, A., 1989, Udang Renik Air Asin (Artemia salina), 15-18, Penerbit Bhatara Karya Aksara, Jakarta.
Mudjiman, A., 1991, Makanan Ikan, 72-88, Penebar Swadaya, Jakarta.
Mursyidi, A.,1990, Analisis Metabolit Sekunder, 171-175, UGM Press,Yogyakarta. Nafrialdi dan Ganiswarna S., 1995, Antikanker cit Ganiswara, Farmakologi dan
Terapi, 686, Bagian Farmakologi Fakultas Kedokteran Universitas Indonesia, Jakarta.
Nuswantari, D., 1998, Kamus Saku Kedokteran Dorland, Edisi 25, 900, ECG, Jakarta. Peter, J.H. and Amala, R., 1998, Laboratory Handbook for the Fractionation of
Natural Extracts, 39,118-119, Thomson , London.
Rana, V.S., Prasad, D., Blazquez, M.A., 2005, Chemical Composition of the Leaf Oil
of Lantana camara,
http://www.findarticles.com/p/articles/mi_qa409/is_200503/ai_n13505544. Diakses pada 20 Februari 2006.
Robbers, JE., Speedie, MK., Tyler, VE., 1996, Pharmacognosy & Pharmacognotechnology, 89-90, 138, Lea & Febiger, USA.
Robinson T., 1991, The Organic Constituent of Higher Plants, diterjemahkan oleh Kosasih Padmawinata dan Iwang Soediro, 115, ITB Press, Bandung.
Schunack, W., Mayer, K., Haake, M., 1990, Senyawa Obat, Buku Pelajaran Kimia Farmasi, Edisi kedua, 780-791, UGM Press, Yogyakarta.
Sharma, O.M.P., and Sharma, P.D., 1989, Natural products of the Lantana plant- the present and prospects, Journal of Scientific Industrial Research, Vol 48, 471-478.
Soelastru, 1986, Penelitian Farmakognosi dan Kandungan Kimia dari daun Lantana camara , Skripsi, Fakultas Farmasi Unair, Surabaya.
Solis, P.N., Wright, C.W., Anderson, M.M., Gupta, M.F., Philipson, J.D., 1993, A Microwell Cytotoxicity Assay using Artemia salina (Brine Shrimp), Planta Medica, 59,250-252.
Stahl, 1985, Drug Analysis by Chromatography and Microscopy, diterjemahkan oleh Kosasih Padwawinata dan Iwang Soediro, 3-17, ITB Press, Bandung.
Tyler, VE., Brady, LR., Robbers, JE., 1988, Pharmacognosy, 9th edition ,443-444, Lea&Febiger, Philadelphia.
Wagner, H., Brady, S., dan Zgainski, E. M., 1984, Plant Drug Analysis A Thin Layer Chromatography Atlas, 164, 226, Springer-Verlag, Berlin.
Lampiran 2. Foto tumbuhan tembelekan
Lampiran 4. Orientasi untuk mendapatkan seri konsentrasi yang akan digunakan dalam pengujian
A. Pembuatan larutan A dan larutan B 1. Larutan A (10 mg/ml)
Larutan A dibuat dengan menimbang 100 mg ekstrak etanol daun tumbuhan tembelekan kemudian dilarutkan dalam etanol sampai 10 ml.
2. Larutan B (1 mg/ml)
Larutan B dibuat dengan mengambil 1 ml dari larutan A kemudian dilarutkan dalam etanol sampai 10 ml.
B. Pembuatan larutan dengan konsentrasi 10,100,1000 μg/ml
Dari larutan B (1 mg/ml), dibuat seri konsentrasi 10 dan100 μg/ml. 1. Konsentrasi 10 μg/ml = 0,01 mg/ml V1 x C1 = V2 x C2 V1 x 1 mg/ml = 5 ml X 0,01 mg/ml V1 = ml mg ml mg mlx / 1 / 01 , 0 5 = 0,05 ml 2. Konsentrasi 100 μg/ml = 0, 1 mg/ml V1 x 1 mg/ml = 5 ml X 0, 1 mg/ml V1 = ml mg ml mg mlx / 1 / 1 , 0 5 = 0, 5 ml
Untuk konsentrasi 1000 μg/ml dibuat dari larutan A. 3. Konsentrasi 100 μg/ml = 1 mg/ml
V1 = ml mg ml mg mlx / 10 / 1 5 = 0, 5 ml
C. Jumlah larva artemia yang mati tiap 10 ekor
Perlakuan (μg/ml) Kontrol (μg/ml) Replikasi 10 100 1000 10 100 1000 1 2 9 8 1 1 1 2 3 6 10 0 1 1 3 1 9 10 0 2 2 4 1 8 10 2 2 2 5 3 9 10 1 0 0 % rata-rata 20 82 10 8 12 12 % rata-rata= 10 5 x jumlah
D. Perhitungan % kematian dengan rumus Abbot:
% kematian pada tes uji – % kematian pada kontrol
% kematian = X 100%
100 – % kematian pada kontrol
1. Konsentrasi 10 μg/ml = 100% % 8 100 % 8 % 20 x − − = 13,04% 2. Konsentrasi 100 μg/ml = 100% % 12 100 % 12 % 82 x − − = 79,55% 3. Konsentrasi 1000 μg/ml = 100% % 12 100 % 12 % 96 x − − = 95,45%
Dipilih konsentrasi yang % kematiannya antara 20%-80%, sehingga dipilih konsentrasi terendah yaitu 10 μg/ml dan konsentrasi tertinggi yaitu 100 μg/ml.
E. Penentuan seri konsentrasi F = n−1 /
SD
LD ket: F= faktor pengali
n= jumlah seri konsentrasi yang diinginkan LD = konsentrasi terbesar SD = konsentrasi terkecil F = 5−1100/10 = 410 = 1,8 dibulatkan menjadi F = 2 Seri konsentrasi: 1. Dosis terendah =10 μg/ml = 0,01 mg/ml 2. 10 μg/ml x 2 = 20 μg/ml = 0,02 mg/ml 3. 20 μg/ml x 2 = 40μg/ml = 0,04 mg/ml 4. 40 μg/ml x 2 = 80 μg/ml = 0,08 mg/ml 5. 80 μg/ml x 2 = 160 μg/ml = 0,16 mg/ml
F. Pembuatan larutan dengan konsentrasi 10, 20, 40, 80,dan 160 μg/ml
Dari larutan B (1 mg/ml), dibuat seri konsentrasi 10, 20, 40, 80,dan 160 μg/ml Konsentrasi
(μg/ml)
Jumlah yang diambil dari larutan B (ml) 10 0,05 20 0,1 40 0,2 80 0,4 160 0,8
G. Jumlah larva artemia yang mati tiap 10 ekor Perlakuan (μg/ml) Kontrol (μg/ml) Replikasi 10 20 40 80 160 10 20 40 80 160 1 2 1 2 10 9 0 0 2 3 0 2 1 2 3 5 9 1 2 0 1 0 3 1 4 3 5 10 2 1 0 1 2 4 2 2 2 4 9 0 0 1 1 2 5 2 1 2 6 10 10 0 1 2 0 % rata2 16 20 24 60 94 8 6 8 16 8
H. Data % kematian dengan umus Abbot: Konsentrasi (μg/ml) % kematian larva artemia 10 8,69 20 14,89 40 17,39 80 52,38 160 93,47
Dipilih konsentrasi yang % kematiannya antara 20%-80%, sehingga dipilih konsentrasi terendah yaitu 40 μg/ml dan konsentrasi tertinggi yaitu 100 μg/ml (dari data sebelumnya).
I. Penentuan seri konsentrasi F = 5−1100/40 = 4 2 = 1,26 dibulatkan menjadi F = 1,3 ,5 Seri konsentrasi: 1. Dosis terendah = 40 μg/ml = 0,04 mg/ml 2. 40 μg/ml x 1,3 = 52 μg/ml = 0,052 mg/ml 3. 52 μg/ml x 1,3 = 68 μg/ml = 0,068 mg/ml 4. 68 μg/ml x 1,3 = 88 μg/ml = 0,088 mg/ml 5. 88 μg/ml x 1,3 = 114 μg/ml= 0,114 mg/ml
J. Pembuatan larutan dengan konsentrasi 40, 52, 68, 88, dan 114 μg/ml
Dari larutan B (1 mg/ml), dibuat seri konsentrasi 40, 52, 68, 88, dan 114 μg/ml. Konsentrasi
(μg/ml)
Jumlah yang diambil dari larutan B (ml) 40 0,2 52 0,26 68 0,34 88 0,44 114 0,57
Lampiran 5. Jumlah kematian larva artemia akibat pemberian ekstrak etanol daun tumbuhan tembelekan
A. Jumlah larva artemia yang mati tiap 10 ekor
Perlakuan (μg/ml) Kontrol (μg/ml) Replikasi 40 52 68 88 114 40 52 68 88 114 1 4 5 6 8 9 2 1 1 0 1 2 3 5 5 9 9 1 0 0 1 2 3 5 6 7 6 8 0 2 2 0 0 4 4 3 6 8 8 0 2 1 1 0 5 4 3 5 7 7 2 1 2 1 1 % rata2 40 44 58 76 82 10 12 12 6 8
B. Data % kematian dengan umus Abbot: Konsentrasi (μg/ml) % kematian larva artemia 40 33,33 52 36,36 68 52,27 88 74,47 114 80,43
Lampiran 6. Perhitungan data statistik SPSS 10.00 dengan menggunakan analisis probit terhadap ekstrak etanol daun tumbuhan tembelekan
* * * * * * * * * * * * P R O B I T A N A L Y S I S * * * * * * * * * * * *
Parameter estimates converged after 12 iterations. Optimal solution found.
Parameter Estimates (PROBIT model: (PROBIT(p)) = Intercept + BX): Regression Coeff. Standard Error Coeff./S.E.
KONS 3,14850 1,19378 2,63742 Intercept Standard Error Intercept/S.E. -5,60866 2,18159 -2,57090 Pearson Goodness-of-Fit Chi Square = ,411 DF = 3 P = ,938
Since Goodness-of-Fit Chi square is NOT significant, no heterogeneity
factor is used in the calculation of confidence limits.
- - - - - -
* * * * * * * * * * * * P R O B I T A N A L Y S I S * * * * * * * * * * * *
Observed and Expected Frequencies
Number of Observed Expected
KONS Subjects Responses Responses Residual Prob 1,60 10,0 3,3 2,862 ,468 ,28618 1,72 10,0 3,6 4,185 -,545 ,41847 1,83 10,0 5,2 5,640 -,410 ,56395 1,94 10,0 7,5 6,962 ,488 ,69622 2,06 10,0 8,0 8,072 -,032 ,80717
* * * * * * * * * * * * P R O B I T A N A L Y S I S * * * * * * * * * * * *
Confidence Limits for Effective KONS
95% Confidence Limits Prob KONS Lower Upper ,01 11,02806 ,06013 24,31493 ,02 13,46107 ,13024 27,35107 ,03 15,27609 ,21257 29,48458 ,04 16,80102 ,30719 31,20799 ,05 18,15304 ,41438 32,69158 ,06 19,38919 ,53449 34,01649 ,07 20,54213 ,66803 35,22843 ,08 21,63250 ,81555 36,35593 ,09 22,67432 ,97767 37,41822 ,10 23,67762 1,15508 38,42890 ,15 28,32633 2,29954 42,99260 ,20 32,66365 3,96209 47,15237 ,25 36,91030 6,29519 51,23170 ,30 41,19245 9,49435 55,46560 ,35 45,60274 13,79763 60,11821 ,40 50,22352 19,46160 65,59505 ,45 55,13934 26,66786 72,64694 ,50 60,44675 35,28154 82,78312 ,55 66,26503 44,52797 98,88722 ,60 72,75098 53,24769 125,49553 ,65 80,12260 60,92250 168,80695 ,70 88,70098 67,87619 238,65601 ,75 98,99161 74,69383 354,11661 ,80 111,86165 81,98566 556,94620 ,85 128,98990 90,52534 953,17663 ,90 154,31494 101,75735 1888,61125 ,91 161,14308 104,58879 2229,55907 ,92 168,90375 107,72613 2670,74866 ,93 177,86913 111,25449 3258,14747 ,94 188,44574 115,29880 4069,31317 ,95 201,27820 120,05338 5245,34205 ,96 217,47547 125,84534 7070,69846 ,97 239,18497 133,29198 10211,44677 ,98 271,43535 143,79245 16654,74387 ,99 331,31939 161,88143 36043,79924
Probit Transformed Responses
Log of KONS 2,1 2,0 1,9 1,8 1,7 1,6 1,5 P robi t 1,0 ,8 ,6 ,4 ,2 -,0 -,2 -,4 -,6 Rsq = 0,9485Lampiran 7. Foto kromatogram identifikasi triterpenoid
A B C
Keterangan:
Fase diam : silika gel GF 254 Fase gerak : toluene:etil asetat (93:7) Deteksi : A. Visibel
B. UV 254 nm
C. UV 365 nm
1. Sampel : ekstrak etanol daun tumbuhan tembelekan 2. Pembanding : ekstrak etanol Liquiritiae Radix
Lampiran 8. Jurnal penggunaan uji Brine Shrimp Lethality Test
Jou r n a l of D e n t ist r y
Volum e 28, I ssue 5 , June 2000, Pages 341- 345 doi: 10.1016/ S0300- 5712( 00) 00007- 5
Copyr ight © 2000 Elsev ier Science Lt d. All r ight s reserved
A new screening test for toxicity testing of dental
materials
M. Pelka, C. Danzl, W. Distler and A. Petschelt
Policlinic for Operative Dentistry and Periodontology, University of Erlangen-Nuremberg, Glueckstr. 11, D-91054 Erlangen, Germany
Received 7 May 1999; revised 16 August 1999; accepted 29 November 1999. Available online 24 April 2000.
Abstract
Objectives: The development of a micro plate assay for cytotoxicity testing of dental materials based on a bioassay using brine shrimp larvae (artemia salina) as sensitive organisms.
Methods: Brine shrimp larvae are commonly used for cytotoxicity assays in
pharmacology. These larvae are sensitive to toxic substances. The ratio between dead larvae (no motility) and living larvae (high motility) in comparison to a control without any toxic substances is used to estimate the toxicity of the test solutions. The test materials (Arabesk®, Solitaire®, Pertac® II, Tetric®, Herculite® and the compomer materials Dyract®, Hytac®, Compoglass®) were polymerized and
consecutively milled. After incubation of 1 g in 4 ml distilled water at 37°C for 48 h, the solid materials were separated by centrifugation. The solutions were equibrilated with NaCl to a salt content of 25 g/l. Aliquots of 200 μl were distributed in eight micro wells and 50 μl of a artemia salina containing (n=8–14) solution were added to each well. As controls eight wells with 250 μl salt solution containing a comparable number of brine shrimp were used. At baseline, after 2, 5, 24 and 48 h, the dead shrimp were counted using a stereo microscope. Finally all shrimps were sacrificed using Na-acid (5%) and counted to get the number of shrimps per well.
Results: All compomers and Solitaire caused 100% brine shrimp lethality after 24 h and showed significantly (p<0.01, signed rank test) higher toxicities than the remaining composites. With the exception of Pertac II, all composites showed
significantly higher toxic values than the control. Pertac II did not show any differences from the controls used.
Conclusions: This new technique has some advantages for toxicity testing of restorative materials, because it can quickly be carried out at low costs. The disadvantage is the high quantity of material used and the low sensitivity. Author Keywords: Elution; Monomer; Cytotoxicity; Artemia salina
Corresponding author. Tel.: +49-9131-8536310; fax: +49-9131-8533603; email: [email protected]
Journal of the Chilean Chemical Society
ISSN 0717-9707 versi on-line
SECONDARY METABOLITES FROM FOUR MEDICINAL PLANTS FROM NORTHERN CHILE: ANTIMICROBIAL ACTIVITY AND BIOTOXICITY AGAINST Artemia salina.
GLAUCO MORALES * , PATRICIA SIERRA, ARLETT MANCILLA, ADRIN PAREDES, LUIS A. LOYOLA, OSCAR GALLARDO AND JORGE BORQUEZ
Laboratorio de Productos Naturales , Departamento de Qu ca , Facultad de Ciencias B cas , Universidad de Antofagasta, Casilla 170 . Antofagasta , Chile.
( Received : June 20, 2001 Accepted : October 15,2002 ) ABSTRACT
Antibacterial activity and biotoxicity against Artemia salina of chloroform and alcohol extracts and isolated products from four plants used in ethnomedicine in northern Chile is reported. Nine compounds already identified were isolated from aerial parts of Artemisia copa Phil., Acantholippia punensis Botta, Ephedra andina Poepp. ex C. A . Mey and Haplopappus rigidus Phil : 3,5 dihydroxy 6, 7, 3', 4' tetramethoxyflavone, lupeol, β amyrine , β sitosterol, ephedrine, 2
ethylhexanol phthalate , 18 acetoxy cis cleroda 3,13 Z dien 15 oic acid, 5,4' dihydroxy 7 methoxyflavanone and 3,5,7 trihydroxy 6, 4' dimethoxyflavone. Key Words: Artemisia copa Phil.(copa-copa), Acantholippia punensis Botta. (rica rica) , Ephedra andina Poepp.ex C. A . Mey (pingo pingo), Haplopappus rigidus
Phil. (baylahuen) , medicinal plants, flavonoids, terpenoids, antimicrobial activity INTRODUCTION
Andean High Plateau in Northern Chile (Puna Atacame between 3,000 to 4,200 m above sea level, is an ecoregion characterized by a very low relative humidity, cloudless skies during most of the year, daily large temperature extremes and between summer and winter, and a rainfall season in summer known as the Altiplano Winter. Small and distant villages are present in this particular
ecological system. Ollague (2113' S, 6827' W) in the north and Socaire (2336' S, 6750' W) in the south, have a combined population of nearly 5,000 inhabitants1. This report provides information on the antimicrobial properties and toxicity against Artemia salina of alcohol and chloroform extracts and secondary
metabolites isolated from four medicinal plants used by Atacame communities 1
-8
: Artemisia copa Phil., ( Asteraceae ), Acantholippia punensis Botta ( Verbenaceae), Ephedra andina Poepp.ex C. A . Mey ( Ephedraceae) and
Haplopappus rigidus Phil ( Asteraceae).
Microbial resistance to antibiotics in use nowadays provides the need for the search of new compounds with potential effects against pathogenic bacteria 9-10. Infusions of Artemisia copa ( copa copa) are used against abdominal, liver and kidneys pains. Acantholippia punensis ( rica rica) infusions are used against colds, stomach and liver discomforts and to improve blood circulation. Ephedra andina ( pingo pingo ) infusions are used against asthma, liver and urinary bladder discomforts and as antitussive and decongestant alternative. Haplopappus rigidus ( bailahu鮠) infusions are used to prevent or as a cure of liver and
gastrointestinal disorders and as a sexual stimulant. RESULTS AND DISCUSSION
Shows the results obtained after the evaluation of the antimicrobial activity of four medicinal plants extracts from Northern Chile :Artemisia copa Phil., Acantholippia punensis Botta , Ephedra andina Poepp.ex C. A . Mey and Haplopappus rigidus Phil. and 3 isolated products.
Inhibition zone measurements using the filter paper disks method showed that chloroform and alcohol extracts have interesting effects against Gram positive bacteria (Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC 29212), Bacillus subtilus (ATCC 6633)). Their effects were negligible against Gram negative bacteria (Acinetobacter baumanni (ATCC 19606), Salmonella typhi (ATCC 3492), Escherichia coli (ATCC 25922), Pseudomona aeruginosa (ATCC 27853)) and
Candida albicans (ATCC 10231). Growth of Staphylococcus aureus and Bacillus subtilus was affected by the four plants extracts and the effect was much higher on
Bacillus subtilus. Only extracts from Acantholippia punensis (rica rica) showed inactivation of Enterococcus faecalis. Artemisia copa ( copa copa) , used against stomach, liver and spleen discomforts, was active against 7 out of 8 strains used in this study, including Candida albicans. These results could be related to the used of these plants as a remedy against urinary and intestinal infections by the Atacame people; however, isolated, purified products were inactive. Compound 6, (2 - ethyl) hexanol phthalate, showed the highest activity against S. aureus, B. subtilus and C.albicans. β - sitosterol ( Compound 4) and β - amyrine showed a marginal activity against E. coli.
The four plants extracts were highly toxic to Artemia nauplii and LD50 values ranged from 0.023 to 7.29 mg / mL. Ephedra andina ( pingo pingo) extract was the most toxic. Only three compouds showed biotoxicity levels below 300 mg/mL and b -amyrine had the highest activity.
Compound 1 was isolated from Artemisia copa and its flavonoid nature was confirmed by its physical and spectroscopic properties. Its molecular formula, C 19H 18 O 8 , based on mass spectroscopy, is in agreement with the information inferred from the NMR spectra, that is, four methyl groups and two OH groups, one of them
located at C 5 ( d 12.65). The flavonol nature comes from the resonance values of C 2 , C 3 and C 4 carbon atoms at ring C, δ 148.25 , 138.61 and 178.80, respectively, in agreement with this flavonoid class. This information matches with that from 3,5 dihydroxy 6,7,3',4' tetramethoxyflavone 15.
Acantholippia punensis ( rica rica ) provided us with two isomeric, triterpenic natural products ( M+ 426). Compound 2 showed spectroscopic and physical properties identical to those described for 3β - hydroxylup- 20(29) en , known as lupeol 16-17 . According to the available data, Compound 3 is a 3β - hydroxyolean 12 - en known as β -amyrine 18.
From Ephedra andina ( pingo pingo) we isolated three products . Compound 4, with a molecular formula C 29H50 O , showed physical and spectroscopic properties identical to those described for b - sitosterol ( 3β stigmast 5 en 3 ol )19 - 20 . Compounds 5 and 6 were obtained from an extraction protocol for alcaloids isolation. Compound 5 was identical in all properties to a sample of () ephedrine , a well known antitussive and decongestant . Compound 6 was an aromatic ester (C24H34 O4 , 1730, 1600 , 1580 cm-1). NMR spectra were central on structure determination since they showed a highly symmetrical molecule. The aromatic signals between d 7.70 and 7.52 ppm on the 1H NMR spectra have reazonable coupling constants for protons at the ortho- substituted ring. Signal at d 4.23 ppm is assigned to a methylene group geminal to the ester alcohol group. 2D NMR experiments allowed us to propose (2 - ethyl) hexanol phthalate as its molecular formula. This compound has been recently isolated as a natural product from Cassia auriculata 21.
Crystaline Compound 7 was isolated from Haplopappus rigidus (bailahu鮩 along other previously reported compounds: rigiduside (13 O β xylopiranosil 13 epimanool) 22, rigidusol (13 hydroxy 18 acetoxy cis clerode 3,14 dien) and desacetylrigidusol ( 13,18 dihydroxy cis clerode 3,14 dien)23.Its physical and spectroscopic properties indicated the presence of an ester and an acid group, both a, b unsaturated, which coincided with those described for 18 acetoxy cis clerode 3,13 Z dien 15 oic acid , a product we have isolated from Croton chilensis and its structure was confirmed by X rays crystalographic techniques24.
Properties of Compound 8 indicated it could be a flavonoid. Its uncolored crystals, optical activity and NMR spectra suggest a flavonone structrure and, considering the whole information available, its structure would be 5,4' dihydroxy 7 metoxyflavanone 25.
Finally, properties of Compound 9 are in agreement with those from 3,5,7 trihydroxy 4',6 dimethoxyflavone 26
© 2007 Sociedad Chilena de Qu ca
Paicav 70, Depto. 19 P.O. Box 2613, Concepci Chile
BIOGRAFI PENULIS
Natalia Sugianti dilahirkan di Purwokerto pada tanggal 16 Desember 1984 sebagai putri pertama dari pasangan Jong Sugiyanto Setiyawan dan Lidia Setiati. Penulis menempuh pendidikan di Taman Kanak-Kanak Karitas pada tahun 1989-1991, dan pada tahun 1997 menyelesaikan pendidikan di Sekolah Dasar Karitas. Pada tahun 1997-2000, penulis menempuh pendidikan di Sekolah Menengah Pertama Negeri 8 Purwokerto, dan melanjutkan ke Sekolah Menengah Umum Negeri 1 Purwokerto pada tahun 2000-2003. Pada tahun 2003, penulis melanjutkan pendidikan ke jenjang Strata Satu (S1) Fakultas Farmasi Universitas Sanata Dharma Yogyakarta. Selama kuliah penulis pernah menjadi panitia Pekan Ilmiah Mahasiswa Farmasi Indonesia (PIMFI) dan panitia Lokakarya Kurikulum Program Profesi Universitas Sanata Dharma. Selain itu penulis juga pernah menjadi asisten Kimia Analisis (2006).