1 EFEK TOKSISITAS KOMPONEN BIOAKTIF DAUN LOBAK (Raphanus sativus Landra. var. hortensis Back.) DENGAN METODE BRINE SHRIMP LETHALITY TEST SEBAGAI KANDIDAT ANTIKANKER DAN PROFIL KROMATOGRAFI LAPIS TIPISNYA TOXICITY EFFECT BIOACTIVE COMPOUND OF LOBAK (Raphanus sativus Landra. var. hortensis Back.) LEAVES WITH BRINE SHRIMP LETHALITY TEST METHOD AS ANTICANCER CANDIDATE AND THAT THIN LAYER CRHOMATOGRAPHY PROFILE Yustin Nur Khoiriyah, Rita Rakhmawati, Endang Anggarwulan Department of Biology, Faculty of Mathematics and Natural Sciences, Sebelas Maret University, Surakarta. ABSTRACT Cancer have a great rank position to cause of people death in developing country (Indrayani et al., 2006). The synthetic drug of cancer to healing trade have been give side effects and almost expensive relatively. This reason is push ahead to discover a new source drug of cancer. Indonesia have a great potency in this case. One of the plant which have potency to be a new source drug of anticancer candidate is Lobak (Raphanus sativus L.). This vegetables as drug usefull have various bioactive compound on this leaves and tuber, and it has been used empirical for anticancer drugs. The new source drug as a candidate of anticancer require to be proved scientifically. In this research was conducted toxicity effect from bioactive compound of Lobak (Raphanus sativus L.) leaves with BST method as pre screening to discover an anticancer compound. The brine shrimp lethality test as toxicity test has been done by including ten larva A. salina Leach into flacon contain test sampel. Death percentage of A. salina Leach larva was counted 24 hours after giving of test sampel rate series, then made equation of linear line to determine values of LC 50-24 hours . The result of toxicity test showed that methanol extract more toxic than chloroform extract. The partition result of methanol extract showed that insoluble fraction of ethyl acetate more toxic than soluble fraction of ethyl acetate. Insoluble fraction of acetate ethyl as toxic fraction, then be partitionated with acetonitril. The test result of partition with acetonitril by BST showed that insoluble fraction acetonitril is more toxic than soluble fraction of acetonitril with values of LC 50-24 hours = 90,54 g/mL. TLC chromatogram with various specific reagent showed that insoluble fraction of acetonitril found a phenolic compound. Keyword : Raphanus sativus L. var hortensis Back, toxicity, BST, and anticancer
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EFEK TOKSISITAS KOMPONEN BIOAKTIF DAUN … saponin, flavonoid dan polifenol, minyak atsiri, vitamin A dan C serta bijinya mengandung 30-40% minyak lemak dan minyak atsiri. Zat-zat
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EFEK TOKSISITAS KOMPONEN BIOAKTIF DAUN LOBAK (Raphanus
sativus Landra. var. hortensis Back.) DENGAN METODE BRINE SHRIMP
LETHALITY TEST SEBAGAI KANDIDAT ANTIKANKER DAN PROFIL
KROMATOGRAFI LAPIS TIPISNYA
TOXICITY EFFECT BIOACTIVE COMPOUND OF LOBAK (Raphanus
sativus Landra. var. hortensis Back.) LEAVES WITH BRINE SHRIMP
LETHALITY TEST METHOD AS ANTICANCER CANDIDATE AND
THAT THIN LAYER CRHOMATOGRAPHY PROFILE
Yustin Nur Khoiriyah, Rita Rakhmawati, Endang Anggarwulan
Department of Biology, Faculty of Mathematics and Natural Sciences,
Sebelas Maret University, Surakarta.
ABSTRACT
Cancer have a great rank position to cause of people death in developing
country (Indrayani et al., 2006). The synthetic drug of cancer to healing trade have
been give side effects and almost expensive relatively. This reason is push ahead
to discover a new source drug of cancer. Indonesia have a great potency in this
case. One of the plant which have potency to be a new source drug of anticancer
candidate is Lobak (Raphanus sativus L.). This vegetables as drug usefull have
various bioactive compound on this leaves and tuber, and it has been used
empirical for anticancer drugs. The new source drug as a candidate of anticancer
require to be proved scientifically.
In this research was conducted toxicity effect from bioactive compound of
Lobak (Raphanus sativus L.) leaves with BST method as pre screening to discover
an anticancer compound. The brine shrimp lethality test as toxicity test has been
done by including ten larva A. salina Leach into flacon contain test sampel. Death
percentage of A. salina Leach larva was counted 24 hours after giving of test
sampel rate series, then made equation of linear line to determine values of LC50-24
hours.
The result of toxicity test showed that methanol extract more toxic than
chloroform extract. The partition result of methanol extract showed that insoluble
fraction of ethyl acetate more toxic than soluble fraction of ethyl acetate. Insoluble
fraction of acetate ethyl as toxic fraction, then be partitionated with acetonitril.
The test result of partition with acetonitril by BST showed that insoluble fraction
acetonitril is more toxic than soluble fraction of acetonitril with values of LC50-24
hours = 90,54 �g/mL. TLC chromatogram with various specific reagent showed that
insoluble fraction of acetonitril found a phenolic compound.
Keyword : Raphanus sativus L. var hortensis Back, toxicity, BST, and anticancer
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PENDAHULUAN
Kanker menempati peringkat tertinggi sebagai penyebab kematian di
negara berkembang (Indrayani et al., 2006). Usaha penyembuhan dengan obat
kanker sintetis umumnya masih relatif mahal dan memiliki efek samping yang
besar. Hal tersebut mendorong dilakukannya pencarian sumber obat baru yang
berasal dari alam sebagai salah satu kandidat yang berkhasiat antikanker.
Salah satu tanaman yang berpotensi sebagai sumber obat baru untuk
kandidat antikanker adalah tanaman Lobak (Raphanus sativus L.). Tanaman sayur
berkhasiat obat ini, umbi dan daunnya mengandung berbagai kandungan kimia
seperti saponin, flavonoid dan polifenol, minyak atsiri, vitamin A dan C serta
bijinya mengandung 30-40% minyak lemak dan minyak atsiri. Zat-zat tersebut
mengandung antibiotik terhadap beberapa jenis bakteri dan antioksidan (Mursito,
2007; Kumalaningsih, 2008). Beberapa penelitian Lobak telah dilaporkan tentang
ekstrak metanol 95% akar Lobak segar, telah diisolasi satu senyawa penangkap
radikal DPPH dengan EC50 0,701 mg/mL yang diduga sebagai turunan
isotiosianat (Ervina et al., 2001). Senyawa isotiosianat sendiri diketahui sanggup
berperan memblok aktivasi pembentukan metabolit karsinogen dan meningkatkan
detosifikasi karsinogen (Yulianto, 2005), sehingga berkhasiat sebagai
hepatoprotektor dan mencegah kanker.
Lee et al., (2006) melaporkan hasil penelitiannya bahwa ekstrak metanol
kecambah Lobak mampu menginduksi Quinon Reductase (QR) yang berperan
penting untuk melindungi dari karsinogen dan xenobiotik lain. Zat aktif Indole-3-
carbinolnya, berkhasiat anti tumor, khemopreventif terhadap tumor payudara,
menghambat karsinogenesis terhadap cell line estrogen responsive,
imunomodulator dan mampu meningkatkan produksi TNF (Tumor Necrosis
Factor) sehingga sangat berguna dalam pencegahan terhadap tumor (Weng et al.,
2008). Serat di dalamnya dapat mencegah risiko kanker usus besar (kolon)
(Raharjo, 2004).
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Berdasarkan hal tersebut, Lobak berpotensi sebagai sumber obat baru
kandidat antikanker, namun agar dapat dipertanggungjawabkan, diperlukan
penelitian ilmiah lebih lanjut tentang komponen bioaktif dari daun Lobak.
Uji toksisitas sebagai skrining awal senyawa bahan alam yang diduga
berkhasiat antikanker yaitu dengan menggunakan hewan uji Artemia salina
Leach. yang dikenal dengan Brine Srhimp Lethality Test (BST) (McLaughlin et
al., 1998). Senyawa toksik pada BST mempunyai korelasi terhadap uji sitotoksik,
mengingat sifat toksik terhadap sel (sitotoksik) harus dimiliki suatu senyawa
kandidat antikanker. Penelitian ini dilakukan untuk mengetahui adanya efek
toksisitas dari komponen bioaktif daun Lobak dengan metode BST sebagai
skrining awal pencarian senyawa antikanker.
BAHAN DAN METODE
Alat dan Bahan
Alat-alat yang diperlukan dalam penelitian ini adalah rotary evaporator