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Abstract. Neem leaves (Azadirachta indica A. Juss.) have been traditionally used as antibacterial agent for a long time.
Former research mentioned that neem leaves were proven to have antibacterial activity. This study was aimed to isolate
antibacterial components in the chloroform extract of neem leaves guided by bioautography. Neem leaves were extracted
by maceration technique. Antibacterial testing of chloroform extract was conducted by agar dilution method. Chloroform
extract at the concentration of 1000 µg/mL inhibited the growth of Staphylococcus aureus but not Escherichia coli. The
extract was then fractionated by Vacuum Liquid Chromatography (VLC) method, and the fractions obtained were tested
for antibacterial activity. The mixture of 2 and 3 fractions which was eluted with n-hexane: ethyl acetate = 9:1 (v/v) and
n-hexane: ethyl acetate = 5:1 (v/v) inhibited the growth of S. aureus at the concentration of 1000 µg/mL. TLC-
bioautography of the fraction showed two inhibition zone with hRf values of 43.75 and 18.75. Isolate with hRf of 18.75
has Minimum Inhibitory Concentration (MIC) value of 500 µg/mL and still contained many components such as terpenoid.
Key words: neem, Azadirachta indica A. Juss., antibacterial agent, Staphylococcus aureus.
1 2 3 4 5 6 7 8
Salah satu fraksi aktif diisolasi komponen
aktifnya dengan metode bioautografi. Kontrol Gambar 3. Profil KLT isolat dengan hRf 18,75.
negatif digunakan untuk memastikan bahwa zona Keterangan: Fase diam: silika gel F 254. Fase gerak n-
jernih yang terjadi disebabkan oleh senyawa yang heksana: etil asetat = 1:1 (v/v) + 3 tetes asam asetat
terdapat dalam kromatogram sampel. Hasil uji glasial: 1. UV 254, 2. UV 365, 3. uap amonia, 4.
menunjukkan adanya dua zona jernih dengan hRf Dragendorff, 5. FeCl3, 6. sitroborat, 7. serium (IV) sulfat,
8. anisaldehida-asam sulfat.
46 Biofarmasi 3 (2): 43-46, Agustus 2005
Tabel 4. Hasil analisis KLT isolat daun mimba. Fase diam: DAFTAR PUSTAKA
silika gel F 254, fase gerak n-heksana: etil asetat = 1:1
(v/v), + 3 tetes asam asetat glasial. Backer, C.A. and R.C. Bakhuizen van den Brink Jr. 1965.
Flora of Java (Spermatophytes only). Vol. II.
Penampak bercak Groningen: N.V.P. Noordhoff.
hRF
254 365 UA SB DD FeCl3 CS AA Fabry, W., P.O. Okemo, and R. Ansorg. 1998. Antibacterial
activity of East African medicinal plants. Journal of
18,75 pemadaman - - - - - coklat -
Ethnopharmacology 60 (1): 79-84.
28,75 pemadaman - - - - - kuning -
Gruenwald, J., T. Brendler, and C. Jaenicke. 1998.
48,75 pemadaman - - - - - kuning -
Physicians’ Desk Reference for Herbal Medicines. 1st
56,25 - - - - - coklat hijau
edition. Montvale, NJ.: Medical Economic Company.
68,75 - - - - - coklat ungu
Heyne, A. 1987. Tumbuhan Berguna Indonesia. Jilid III.
78,13 - - - - - - hijau
Jakarta: Badan Litbang Kehutanan.
88,75 - - - - - coklat hijau
Jawetz, Z.E., J.L. Melnick, and E.A. Adelberg. 1986.
98,75 pemadaman - - - - - coklat hijau ungu
Mikrobiologi untuk Profesi Kedokteran. Edisi XIV.
Keterangan: 254: UV 254, 365: UV 365, DD: Dragendorff, Penerjemah: Tonang, H. Jakarta: EGC Penerbit Buku
CS: serium(IV) sulfat, AA: anisaldehida-asam sulfat, UA: Kedokteran.
uap amonia, SB: sitroborat. Mardisiswojo, S. dan H. Rajakmangunsudarso. 1985. Cabe
Puyang Warisan Nenek Moyang. Cetakan Pertama.
Jakarta: Balai Pustaka.
KESIMPULAN Mitscher, L.A., R.P. Leu, M.S. Bathala, W. Wu, J.L. Beal,
1972. Lloydia 35: 157.
Pramularsih, E.D. 2001. Uji Aktivitas Antibakteri Daun
Ekstrak kloroform (konsentrasi 1000 µg/mL) Mimba (Azadirachta indica Juss.) terhadap
aktif menghambat pertumbuhan S. aureus tetapi Staphylococcus aureus dan Salmonella typhi
tidak aktif terhadap E. coli. Isolat pita bawah pada beserta Profil KLTnya. [Skripsi]. Yogyakarta: Fakultas
sistem KLT preparatif dengan fase gerak n-heksana: Farmasi Universitas Gadjah Mada.
etil asetat = 2:1 (v/v) aktif terhadap S. aureus Sudarsono, D. Gunawan, S. Wahyuono, I.A. Donatus, dan
sampai kadar 500 µg/mL. Isolat yang diperoleh Purnomo. 2002. Tumbuhan Obat II, Hasil Penelitian,
mengandung senyawa terpenoid. Sifat-Sifat, dan Penggunaan. Yogyakarta: Pusat Studi
Obat Tradisional UGM.