Cytotoxicity Analysis of Compound A.E. IV.3.4 Isolated From Laban Abang (Aglaia elliptica) Leaf.

Fery Azis Wijaya1,, Mila Trisnoviani2), Nurhadi 3), Dhian Wijaya 3),Prasetyawan Yunianto2,3,5)*, Bambang Srijanto 4), 1) Laboratorium of Cell culture LAPTIAB BPPT, Serpong, Indonesia 2) Laboratorium of chemistry Pharmacy LAPTIAB- BPPT, Serpong, Indonesia
3) Laboratorium of biology Pharmacy- LAPTIAB BPPT, Serpong, Indonesia 4) Central of Technology Pharmaceutical and Medical BPPT, Jakarta, Indonesia 5) Department of Chemistry University of Indonesia, Depok, Indonesia

ABSTRACT
A compound named A.E IV.3.4 was isolated by VLC and column chromatography from ethanolic extract of Laban Abang (Aglaia elliptica) leaf. The isolated compound was tested by by MTT cytotoxic assay using MCF-7 a breast cancer cell and CHO a normal cell, The result showed that A.E.IV.3.4 compound had IC50 of 73 ppm, while cisplatin an anticancer drug had IC50 of 18 ppm, In normal ovary cells A.E.IV.3.4 compound had IC50 of 92 ppm and cisplatin had IC50 of 138 ppm. This indicated that A.E.IV.3.4 compound had moderate toxicity effects in MCF-7 (cancer cell) and had a low toxic effect on CHO cells (normal cell). Cisplatin as a positive control which is commonly used as chemotherapy drugs, had a very toxic effect on MCF-7 cells bu,t not on normal cells. It was concluded that the toxic nature of A.E.IV.3.4 compound had a fairly selective mechanism against cancer cells and was safe in normal cells in similar to cisplatin.

Key word : Aglaia elliptica, chromatography techique, MTT cytotoxic assay. Pendahuluan

Genus Aglaia (Meliaceae) terdiri dari sekitar 130 spesies yang biasanya berbentuk pohon atau semak, tanaman ini tersebar di daerah tropis maupun sub tropis. Secara fitokimia, pada umumnya senyawa yang terkandung dalam Aglaia adalah senyawa Amida dan berdasarkan struktur senyawanya amida tersebut dikelompokkan dalam 2 golongan yaitu : bentuk cyclopenta[bc] benzopyran (=golongan aglain) dan bentuk cyclopenta-[b]benzofuran (=golongan rocaglamide). Beberapa senyawa dari 2 golongan tersebut telah ditemukan dan telah diteliti mempunyai aktivitas sebagai toksisitas terhadap serangga dan anti jamur. Dari beberapa penelitian sebelumnya, dari buah Aglaia elliptica Bl telah diisolasi derivate Rocaglamid oleh Nugroho dkk (1997) sedang pada bagian batang buah telah diisolasi oleh Cui B. dkk (1997) berupa derivat rocaglamide dan dammaran pada bagian batang dan buahnya. Sedangkan Inada dkk (2001) telah mengisolasi dari bagain daun Aglaia elliptica Bl (laban abang) berupa 2 senyawa baru derivate aglaian berupa, 10-O-acetylaglain B (1) dan 4-epiaglain A (2), dua diamides yang sudah diketahui yaitu aglain A (3) dan aminopyrrolidine*)

Correspondence : Laboratorium of Pharmaceutical and Medical BPPT, LAPTIAB Gedung 610 PUSPITEKTANGERANG SELATAN, Tlp 021-7560536, Fax 021-7560536 ext. 122. Email : pyunianto@gmail.com

diamide, odorine, (4), dan tiga senyawa lain yang sudah dikenal dari cycloartanes (5-7) serta Aglain B (8).

Gambar 1. Struktur senyawa hasil isolasi dari daun Aglaia elliptica Bl (laban abang), 10O-acetylaglain B (1), 4-epiaglain A (2), aglain A (3), aminopyrrolidine-diamide, odorine, (4), cycloartanes (5-7), Aglain B (8). Agung Eru (2009) telah melakukan isoalsi dari daun Aglaia elliptica Bl (laban abang) setelah dilakukan proses ektraksi dengan metode maserasi menggunakan etanol, dan pemurnian senyawa menggunakan teknik kromatografi (VLC, kromatografi kolom) dan HPLC preparative dihasilkan senyawa penanda berupa aminopyrrolidinediamide (odorine). Senyawa penanda tersebut dilakukan uji in vitro sitotoksisitas mengunakan metode MTT dengan sel MCF-7, diperoleh hasil daya hambat IC50 terhadap sel MCF-7 sebesar 153.32 ppm. Pada penelitian ini bertujuan untuk mengetahui aktivitas sitotoksisitas dari isolate senyawa dari daun Aglaia elliptica Bl (laban abang), dengan membandingkan aktivitas isolate tarhadap viabilitas sel kanker (MCF-7) dengan sel normal (CHO).
Materials and Methods

Material : leaf of Aglaia elliptica Bl from Bogor Botanical Garden, solvent for ectraxtion : etanol, ethyl acetate, methanol, dichlormetan, isopropanol were destiled from technical grade, Methanol for HPLC (E-Merck), Silica gel 60 (dia 0,040 - 0,063) (E-Merck), Sphadex LH 20 (Sigma).

Methods: Ekstrak etanol laban abang Partisi etanol etanol+air Etil asetat Pemekatan/rotavapor Fraksi etil asetat laban abang Vacuum Liquid Chromatography (VLC) A.E.I . A.E.IVA.E.VIII Silica column (mobile phase = 95 % CH2Cl2 : 5 % 2-C3H7OH) A.E.IV.3 Sphadex column (mobile phase CH3OH) A.E.IV.3.1 Elusidasi : UV spectrum, MS-TOF Toxicicity assay by MTT methods (cell line : MCF-7 and CHO) Heksan

Result and Discussion

AE IV 3.4
K-2800 [1] Pegagan 180

Nam e Retention Time Area

180

160

160

140

S p e k t r u m140U V - V i s A E
1 2 0 0 1 2 0 0

IV 3 .4 m e n it k e 2 2 .6 1 3

120

120

100

1 0 0 0

100 mAU

1 0 0 0

21.517298488 21.753301130 21.978122056 22.07784664 22.218156428 22.6132214357

80

25.068217798 25.432247811 25.727144674 26.02349364 26.443557803

mAU

23.967233700 24.105222735

14.9131601 15.1031997 15.3781520 15.8104401 15.947734 16.1602192 16.4173882 16.5871230 16.8605645 17.0439730 17.48025338 17.8009346 18.0337732 18.33326133 18.67222618 18.97887989 19.39062609 19.71529268 19.90835991 20.227157111 20.56535841 20.68738531 20.79553657 21.01336431

23.420100386

27.262104300 27.54854840 27.81084213

8 0 0

800 8 0

m AU 28.49548924

0.5922178 0.87723571 1.04319408 1.11314089 1.24018694 1.38026243 1.64770903 1.91256741 2.01832590 2.13255322 2.26740429 2.380147396

60

29.27227924 29.647877
250
n m

60
6 0 0 m AU

3.127178858

3.770102128

4.262107175 4.67514483 4.82713045 4.97026961 5.17528769 5.63011137 5.8853385

6 0 0

12.9433578 13.32761 13.443226

12.0201810 12.222127

6.7088749 7.177269 7.522501 7.812138

9.2832260 9.595166 9.715264 9.89877

40

14.008595 14.350656

10.955187

40
4 0 0

4 0 0

20

20
2 0 0

2 0 0

0
20

-20 0 2 4 6 8 10 12 14 M inutes 16 18 20

22

24

26

28

2 2 0 2 10 2 2 3 0 2 4 0 2 5 0 2 6 0 2 7 0 2 8 0 3 90 3 0 3 1 3 2 0 3 0 3 4 0 3 5 0 3 6 0 3 70 3 80 4 9 0 0

30

Chromatogram HPLC of A.E.IV.3.1 with retention time 22.613 min and spectrum UV = 218, 284 nm

333899186

-20

Data TOF MS ES+ of A.E.IV.3.1 with mw about 344 g/mol. Dari data elusidasi terhadap isolate senyawa A.E.IV.3.1 mempunyai spectrum UV = 218, 284 nm,
sedangkan berat molekul sekitar 344 g/mol tapi masih perlu dikomfrimasi lagi, karena isolate belum murni dan perlu dielusidasi dengan NMR. Untuk uji Toksisitas pada MCF-7 dan CHO dapat dilihat pada Tabel berikut :

Sample A.E.IV.3.1 compound Cisplatin (a breast cancer drug)

IC50 Cell Line MCF-7 (breast cancer cell) 73 18 CHO (normal cell) 92 138

This indicated that A.E.IV.3.4 compound had moderate toxicity effects in MCF-7 (cancer cell) and had a low toxic effect on CHO cells (normal cell). Cisplatin as a positive control which is commonly used as chemotherapy drugs, had a very toxic effect on MCF-7 cells bu,t not on normal cells Conclusion The toxic nature of A.E.IV.3.4 compound had a fairly selective mechanism against cancer cells and was safe in normal cells in similar to cisplatin References
i. Agung Eru Wibowo, Karakterisasi Senyawa Utama Ekstrak Daun Aglaia elliptica Blume dan Uji Sitotoksisitas pada Sel Kanker Payudara, http://mru.fk.ui.ac.id/index.php? uPage=profil.profil_detail&smod=profil&sp=public&idpenelitian=5211 Prasetyawan Yunianto, unpblished, Tehnical Report 4 Of 2010 Standardized extract ,

ii.

Program Assement and Aplied Technology of Production Herbal Drug - PTFM, BPPT,

iii. iv. v. vi. vii. viii.

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