Sensitivitas Metode Bioautografi Kontak dan Agar Overlay dalam Penentuan Senyawa Antikapang
Abstract
Bioautography method is a laboratory technique to detect substances inhibiting the growth of tested microorganisms. The sensitivity of contact and agar overlay bioautography Was compared to define a recommended method for detecting antifungal compound. In contact bioautography, n-hexane extract of Alpinia galanga was eluted with dichloromethane and toluene on a thin layer chromatography plate (silica gel GF254). The separated compounds were diffused from chromatogram spots to a Thrichophyton menzagrophytes inoculated Sabouraud Dextrose Agar plate. The plate was incubated at 37°C for 3 days. Inhibition zone was observed and the hRf value was defined. Agar overlay bioautography Was performed by covering chromatogram spots With a molten Sabouraud Dextrose Agar medium seeded by Trichophyton mentagrophytes. After solidification, the chromatogram plate was incubated at 37°C overnight and then was stained with Methyl Thiazole Tetrazolium. The inhibition growth bands were visualized and hRfs value were recorded. A contact bioautography produces oneband while agar overlay bioautography produces four bands. Agar overlay bioautography was therefore more sensitive than contact bioautography.
References
2. Choma I. The use of thin-layer chromatography with direct bioautography for antimicrobial analysis. LCGC Europe. 2005.
3. Colorado RJ, Galeano JE, and Martinez MA. Development of direct bioautography as reference method for testing antimicrobial activity of gentamicin against Escherichia coli. Vitae. 2007, 14(1):67-71.
4. Kavanagh F. Analytical microbiology. Vol Il. New york and London: Academic Press; 1972.p.190-1.
5. Sutrisno RB. Pereaksi KLT (kromatografi lapis tipis). Edisi I. Jakarta: F akultas Farmasi Universitas Pancasila; 1993.
6. Sutrisno RB, Reverse approach. Edisi I. Jakarta: Fakultas Farmasi Universitas Pancasila; 1986.p.13, 24-6,31.
7. Ketaren S. Minyak atsiri bersumber dari bunga dan buah. Bogor: Jurusan Teknologi Jndustri Pertanian. Fakultas Teknologi Pertanian. Institut Pertanian Bogor; 1980. hal.32-6.
8. Tyler VE, Brandy LR, and Robbers JE. Pharmacognosy. 8th ed. Philadelphia: Lea and Febiger; 1997.p.77-81
9. Bobit JM.. Thin layer chromatography. New York: Reinhold publishing Co; 1996.p.900.
10. Stahl E. Analisis obat secara kromatografi dan mikroskopi. Diterjemahkan oleh Padmawinata K dan Sudiro I. Bandung: Penerbit ITB; 1985. hal.3-18.
11. Sastroamidj ojo H. Kromatografi. Yogyakarta: FMIPA
Liberty Press; 1991. hal.26-38.
12. Badaruddin AM. Kromatografi lapis tipis untuk identifikasi alkaloida opium memakai silica gel G. Skripsi. Jakarta: Fakultas Farmasi, Universitas Pancasila; 1976. hal.5-14.
13. Bengue WJ and Kline RM. The use of tetrazolium salt in bioautographic procedures. J Chromatogr. 1972.(664):182-184.
14. Rohalison L, Hamburger M, Hosttetman K, Monod M and Frank E. Abioautography agar overlay method for the detection of antifungal compound from higher plants. Phytochemical Analysis. 1991.(2): 199-203.
15. Fish SA and Codd GA. Bioactive compound production by thermophilic and thermotolerant Cyanobacteria (blue- green algae). World J. Microbiol. Biotechnol. 1994.(10):338-341.
16. Verbitsky SM, McChesney JD, Gourdin GT, Ikenouve LM. Methods and compositions for detecting active components using bioluminescent bacteria and thin layer chromatography. USPTO Patent Application 20070184514. 2006.
17. Runyoro DKB., Mecky IN, Matee MIN, Ngassapa OD, Cosam C Joseph CC and Mbwambo ZH. Screening of Tanzanian medicinal plants for antiCandida activity. BMC Complementary and Alternative Medicine. 2006.(6): 11.
18. Hostettmann K and Marston A. Search for new antifungal compounds from higher plants. Pure &App/. Chern. 1994.(66):2231-4.
19. Homma Y, Sato Z, Hirayama F, Konno K, Shirahama H and Suzui T. Production of antibiotics by Pseudomonas cepacia as an agent for biological control of soilborne plant pathogens. Soil Biol. Biochem. 1989.(21):723-8.
20. Motsei ML, Lindsey KL, Van Staden J, Jager AK. Screening of traditionally used South African plants for antifungal activity against Candida albicans. J.Ethnopharmacol. 2003 Gökalp l`s,cana. 86235-41.
21. Hostettmann K, Marston A, Ndj oko K, Wolfender JL. The potential of African plants as a source of drugs. Curr. Org. Chem. 2000.(4):973-1010.
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