Antibacterial Terpenoid from The Bark of Lansium domesticum Corr cv. Kokossan (Meliaceae).

ABSTRACT

Antibacterial Terpenoid from The Bark of Lansium domesticum Corr cv.
Kokossan (Meliaceae)
Tri Mayanti,1 Siti Soidah 1 , W. Drajat Natawigena,2 Unang Supratman,1 dan Roekmi-ati
Tjokronegoro1
1

Department of Chemistry, Faculty of Mathematics and Natural Sciences , Universitas
Padjadjaran, Jatinangor 45643, Sumedang
2
Department of Plant Diseases and Plant Control, Faculty of Agriculture, Universitas
Padjadjaran, Jatinangor 45643, Sumedang

In the course of our continuing search of novel active compounds from Indonesian plants,
the methanolic extract of bark of L. domesticum cv kokossan showed significant
antibacterial activity against B. cereus and E. coli. The methanolic extract of the bark of
L. domesticum cv kokossan was concentrated and extracted with ethyl acetate. The ethyl
acetate extract exhibited an antibacterial activity with inhibit zone respectively 12.5 and
14 mm against B. cereus and E. coli. The ethyl acetate fraction was separated by
combination of column chromatography on Kieselgel 60 to afford an antibacterial

compound as white crystal with melting point 166-168oC.
Based on UV, IR, H1-NMR, C13-NMR and 2D-NMR spectroscopic evidences and
comparison with previously data reported indicated that isolated compound as terpenoid
group.
An antibacterial compound showed antibacterial activity with inhibit zone 10 mm at
concentration of 1000 ppm against E. coli.

Antibacterial Terpenoid from The Bark of Lansium domesticum Corr cv.
Kokossan (Meliaceae)1
Tri Mayanti,2 Siti Soidah 2 , W. Drajat Natawigena,3 Unang Supratman,2 dan Roekmi-ati
Tjokronegoro2

ABSTRACT

In the course of our continuing search of novel active compounds from Indonesian plants,
the methanolic extract of bark of L. domesticum cv kokossan showed significant
antibacterial activity against B. cereus and E. coli. The methanolic extract of the bark of L.
domesticum cv kokossan was concentrated and extracted with ethyl acetate. The ethyl
acetate extract exhibited an antibacterial activity with inhibit zone respectively 12.5 and 14
mm against B. cereus and E. coli. The ethyl acetate fraction was separated by combination

of column chromatography on Kieselgel 60 to afford an antibacterial compound as white
crystal with melting point 166-168 oC. Based on UV, IR, H1-NMR, C13-NMR and 2DNMR spectroscopic evidences and comparison with previously data reported indicated that
isolated compound as terpenoid group. An antibacterial compound showed antibacterial
activity with inhibit zone 10 mm at concentration of 1000 ppm against E. coli.

Keywords: antibacterial, terpenoid, Lansium domesticum, kokosan, Meliaceae
1

Research artikel was presented at International Confererence On Medicine and Medicinal
Plants, Surabaya, September 8th-9th, 2007.

2

Department of Chemistry, Faculty of Mathematics and Natural Sciences , Universitas
Padjadjaran, Jatinangor 45643, Sumedang
3

Department of Pest Control and Plant Diseases, Faculty of Agriculture, Universitas
Padjadjaran, Jatinangor 45643, Sumedang

INTRODUCTION

Lansium domesticum Corr (Meliaceae) is a popular fruit in southern Asia. The plant
family Meliaceae is noted for the production of useful bitter principles which are generally
used in folk medicine as a drug for diarrhea which caused by bacteria. The bark and leaf
infusion of L. domesticum cv. duku showed antibacterial activity against Echerichia coli,
Staphylococcus aureus, and Bacillus subtiles (Loekitowati and Hermansjah, 2000).
Phytochemical investigation of the fruit peel of L. domesticum revealed the
presence

of

lansic

acid,

3β
-hydroxyonocera-8(26),14-dien-21-one

hydroxyonocera-8(26),14-dien-3-one

(Tanaka

et

al.,

2002).

The

and
seed

21αcontain

tetranortriterpenoids named dukunolides A-F (Nishizawa et al., 1985, 1989). The bark
contain antifeedant triterpenoid namely iso-onoceratriene, 3-keto-22-hydroxyonoceradiene,
onoceradienedione, lansiolic acid, lansiolic acid A and 3-keto lansiolic acid.
In our continuing search on bioactive compounds from Indonesian plants, the

methanolic extract of bark of L. domesticum cv kokossan showed significant antibacterial
activity

against B. cereus and E. coli. In this research, we report the isolation and

characterization of antibacterial triterpenoid, 14-hydroxy-7-onoceradienedione (1).

O

HO

O

(1)

EXPERIMENTAL

General Experimental Procedure. UV and IR spectra were measured with System Perkin
Elmer Spectrum One. 1H and

13

C-NMR spectra were recorded with a JEOL JNM ECA-

500, operating at 500 and 400 MHz (1H), 125 and 100 MHz (13C), using residual and
deutered solvent peaks as internal standards. Vacuum liquid chromatography was carried
out using Kieselgel 60.

Plant Material. Samples of the bark L. domesticum cv kokossan were collected in March
2006 from Cililin, Bandung, Indonesia. The plant was identified by the staff at Department
of Biology, Padjadjaran University.

Bioassay. A 7,2 g nutrient broth (2 g sodium chloride 0,85%; 1,2 g beef extracts and 4 g
pepton) were suspenced in 400 mL hot aquadest which contain 12 g gel. This media was
further sterilized for 15 minutes at 121oC. The bacteria (E. coli and B. Cereus) were
incubated at 37oC for 18-24 hours in this media. Paper disk which contain the sample was
put on the media. The sample was incubated at 37oC for 18-24 hours and inhibit zone was
further measured.

Extraction and Isolation. The dried and milled bark of L. domesticum cv kokossan (3 kg)

was extracted exhaustively by methanol at room temperature. The methanol extract (200 g)
was partitioned between n-hexane and 10% aqueous methanol, and the aqueous phase was
further extracted with ethyl acetate. The ethyl acetate-soluble fraction was subjected to
Kieselgel 60 column eluted with 0-100% dichloro metane/n-hexane. The fraction eluted
with dichloro metane/n-hexane (6:4) contained (1). The fraction was further separated by
Kieselgel 60 column eluted with acetone/n-hexane (1:9) to yield (1) (10,7 mg).
14-hydroxy-7-onoceradienedione (1): colorless amorphous solid, IR (KBr) νmax 3444,
2962-2931, 1705, 1698, 1458, 1338, 1315, 1261-1076, 802 cm-1. 1H-NMR (CDCl3) δH 7.3
(1H, s), 5.4 (1H, s), 2.75, 2.73, 2.70 (2H, dt), 2.6 (2H, m), 2.1 (3H, m), 1,9 ( 2H, m), 1.8
(2H, m), 1.7 (2H, m), 1.6-1.4 (6H, m), 1.3 (3H, s), 1.09 (3H, s), 1.08 (3H, s), 1.05 (3H, s),
1.01 (3H,s), 0.96 (3H, s), 0.92 (3H,s); 13C-NMR (CDCl3) δC see Table 1.

DISCUSSION
Compound 1 was obtained as colorless amorphous solid. The IR spectrum of 1
showed a sharp absorption band at 3444 cm-1 and strong absorption band at 1705 cm-1
indicating the presence of hydroxyl, alkene and ketone groups. The 1H and

13

C-NMR

spectrum of 1 showed the presence of hydroxyl group on C-14 (δC 74,0) (Table 1).
Therefore, it may be concluded that 1 is 14-hydroxy-7-onoceradienedione.

Table 1. 13C-NMR data of 14-hydroxy-7-onoceradienedione (1) on 500 Hz
δH (Σ H,
Posisi C
δC
multiplisitas,
HMBC
COSY
J dalam Hz
1/19
2
3
4/22
5
6
7
8

9
10/18
11/12
13
14
15
16
17

38,54/ 38,37
34,82
216,96
47,62
51,57
28,98
121,70
135,27
55,47
36,58
21,54

61,80
74,03
44,23
31,41
55,23

20

34,13

21
23
24
25
26
27
28
29
30

217, 12
25,14
22,25
13,43
22,33
24,20
15,13
21,43
26,49

2,08; 1,91 (2H,m)

H-2/H-20

2,23; 2,41 (2H, dt, dq, J=15)
1,57 (1H, m)
C-9,C-10,C-24
1,12 (2H, m)
5,41 (1H, m)
C-5, C-9, C-25
1,59 (1H, m)
C-26
1,61 (1H, m)
C-9
1,12 (1H, m)
C-17
1,46 (2H, m)
C-14, C-17
1,51 (2H, m)
C-1,C-4
1,42 (1H, m)
C-18, C-22

2,26;2,73
(2H, dt; td, J=15)

C-19

1,04 (3H, s)
1,08 (3H, s)
0,96 (3H, s)
1,77 (3H, s)
1,21 (3H, s)
0,93 (3H, s)
1,02 (3H, s)
1,09 (3H, s)

20
19
21

11

13

25
1

8

10
3

HO

5
6

H-16
H-15

H-19

O
29

22
17

14 28

30

16

27

O
24

18

26

9

H-6
H-5/H-17
H-26

C-5, C-24
C-3, C-23
C-5, C-9, C-10
C-9
C-13,C-14,C-15
C-13,C-16,C-17
C-17,C-22,C-30
C-17,C-29

12

2

H-1

15

7

23

(1)

Antifungal activity of the methanol, ethyl acetate, and n-hexane extracts of the bark
of L. domesticum kultivar cv. kokosan were tested against B. cereus and E. coli. The
results are tabulated in Table 2, showing that ethyl acetate extracts have inhibit zone 12.5
mm against B. cereus and 14.0 mm against E. coli at 1% w/v. The compound (1) showed

antibacterial activity with inhibit zone 10 mm at concentration of 1000 ppm against E. coli
(Table 3, Fig.1).

Table 2.

Antibacterial activity of the methanol, ethyl acetate, and n-hexane extracts of
the bark of L. domesticum kultivar cv. kokosan with paper disk method.
Inhibit Zone (mm)
Methanol

Bacteria
B. cereus
E. coli

n-hexane

Ethyl acetate

0,1

0,5

1,0

0,1

0,5

1,0

0,1

0,5

1,0

(%)

(%)

(%)

(%)

(%)

(%)

(%)

(%)

(%)

10,5

9,5

12,0

-

-

-

11,0 10,5

12,5

-

13,5

10

-

-

-

9,0

14,0

10,5

Table 3. Antibacterial Activity of Isolate
Inhibit zone (mm)
Bacteria
Acetone
B. cereus
E. coli

8
8

Isolate (500 ppm)
8
8

Isolate (1000 ppm)
8
10

Fig 2. Antibacterial Test of Isolate against B. cereus (a) and E. coli (b).

CONCLUSION
Based on UV, IR, H1-NMR, C13-NMR and 2D-NMR spectroscopic evidences and
comparison with previously data reported indicated that antibacterial compound from the
bark of L. domesticum cv. kokosan is 14-hydroxy-7-onoceradienedione. The compound (1)
showed antibacterial activity with inhibit zone 10 mm at concentration of 1000 ppm
against E. coli.

REFERENCES

Loekitowati, H.P. and Hermansjah. 2000. Studi pemanfaatan biji duku (Lansium
domesticum) untuk obat diare secara in vitro. Jurnal Penelitian Sains. 7: 41-48.
Tanaka, T., M. Ishibashi, H. Fujimoto, E. Okuyama, T. Koyano, T. Kowithayakorn, M.
Hayashi, dan K. Komiyama. 2002. New onoceranoid triterpene constituents from
Lansium domesticum. Journal Natural Product. 65: 1709-11.
Nishizawa, M., Y. Nademoto., S. Sastrapradja., M. Shiro, dan Y. Hayashi. 1985. Structure
of dukunolide, bitter principles of Lansium domesticum. Journal of Organic
Chemistry, 50(26): 5487-90. Washington DC.
Nishizawa, M., M. Emura., H. Yamada., M. Shiro., Chairul, Y. Hayashi dan H. Tozuda.
1989. Isolation of a new cycloartanoid triterpenes from leaves Lansium
domesticum: novel skin-tumor promotion inhibitors. Tetrahedron Letter. 30 (41):
5615-18.