Molekul, Vol. 20. No. 2, July 2025: 394 – 403

Articles

MOLEKUL

eISSN: 2503-0310

https://doi.org/10.20884/1.jm.2025.20.2.16241

Dammarane-Type Triterpenoids from Twigs of Aglaia Foveolata and Their Antibacterial Activity
Ricson Pemimpin Hutagaol1*, Tjandrawati Mozef2, Febi Nurilmala3, Gian Primahana2, Sofa Fajriah2,
Muhammad Eka Prastya2, Sigit Hidayatullah1, Desy Satyaningsih1
Department of Chemistry, Faculty of Mathematics and Natural sciences, Nusa Bangsa University,
Bogor 16166, Indonesia
2
Research Center for Pharmaceutical Ingredients and Traditional Medicine, National Research and Innovation
Agency (BRIN), Serpong Tangerang Selatan 15314, Indonesia
3
Department of Biologi, Faculty of Mathematics and Natural sciences, Nusa Bangsa University,
Bogor 16166, Indonesia
1

*Corresponding author email: ricsonpemimpin70@gmail.com
Received June 12, 2025; Accepted July 12, 2025; Available online July 20, 2025
ABSTRACT. The Aglaia species, which contains triterpenoids, is the most numerous in the Meliaceae family. Aglaia foveolata
(A. foveolata) is a type of plant that has many benefits, as medicinal ingredients. The potential of this plant is inseparable
from the content of various bioactive compounds. This study aims to isolate, characterize the active compound from the twigs
of A. foveolata and test its activity as an antibacterial. Three dammarane-type triterpenoids were isolated from the A.
foveolata twigs which is, namely dammar-24-en-3β,20-diol (1), an epimeric mixture of shoreic and eichlerianic acid (2, 3).
Their chemical structures were determined based on spectroscopic data using infrared, high-resolution mass spectrometry,
and including one and two-dimensional NMR techniques, as well as through data comparison of the reported compound.
Compound 1 was reported for the first time to be successfully isolated from this species. All these substances were tested for
the first time for their antibacterial activity against two Gram-positive bacteria Staphylococcus aureus and Bacillus subtilis
and two Gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa, through this study. Compound 1 was
inactive, the epimeric mixture of 2 and 3 showed moderate antibacterial activity with a minimum inhibitory concentration
(MIC) value ranging from 31.7 to 126.6 ppm, particularly against S. aureus with a MIC value of 31.7 ppm.
Keywords: Aglaia, Elucidation, Isolation, Spectroscopy, Bacterial

INTRODUCTION
Triterpenoids are a structurally diverse group of
natural products that exist in a variety of organisms
exhibiting extensive biological activities (Li et al.,
2023; Ren & Kinghorn, 2019). Aglaia is the
largest genus belonging to the family Meliaceae,
comprising over 130 species distributed mainly in
tropical forest and more than 65 grow in Indonesia
(Mabberley & Pannel, 1995; Pannel, 1992; Pérez et
al., 2014). Various phytochemicals of this genus
have been reported with fascinating bioactivities
including several rocaglate derivatives, triterpenoids,
and steroids (Harneti & Supratman, 2021; Hutagaol
et al., 2021, 2022, 2023). Previous phytochemical
studies of the species A. foveolata reported a
variety of compounds from leaves, bark, and stem
bark, including flavaglines (e.g., silvestrol, bisamides,
and
rocaglamides)
and
dammarane-type
triterpenoids (Pan et al., 2013; Salim et al., 2007).
There are still limited reports regarding antibacterial
activities of triterpenoids from the species of A.
foveolata. An infectious disease is one of the serious

diseases causing high mortality worldwide. For
example, lower respiratory infections and diarrheal
diseases remain the world’s most deadly infectious
diseases, ranked as the fourth and eighth leading
causes of death, respectively. Therefore, the
discovery of new antimicrobial agents, especially
from natural sources in Indonesia, is a very important
subject to study.
In our ongoing efforts to search for triterpenoids
from the Indonesian Aglaia plants, we have further
investigated the twigs of A. foveolata. As a result, three
dammarane-type triterpenoids (1-3) were successfully
isolated and elucidated (Figure 1). Based on literature
search, this is the first time compound 1 has been
successfully isolated from this species. These
triterpenoid compounds were evaluated the
antibacterial activity against Gram-positive bacteria
(Staphylococcus aureus and Bacillus subtilis), and
Gram-negative bacteria (Escherichia coli and
Pseudomonas aeruginosa), which have not been
tested previously for these compounds and the results
are reported in this article.

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Dammarane-Type Triterpenoids

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EXPERIMENTAL SECTION
Materials
Materials the twigs of A. foveolata was obtained
from Timpah Village, Timpah District, Kapuas
Regency, Central Kalimantan, Indonesia, with
coordinate 1.2735° S, 114.5934° E. The voucher
specimen was authenticated by the staff of the
Bogoriense Herbarium, Research Centre for Biology,
Indonesian Institute of Science, Bogor, Indonesia, and
deposited at that herbarium (No. BO-1295312).
Evaluation of antibacterial activity using ATCC
bacteria, including two gram-positive bacteria
(Staphylococcus aureus (S. aureus) ATCC 6538 and
Bacillus subtilis (B. subtilis) ATCC 19659, and two
gram-negative bacteria (Escherichia coli (E. coli) ATCC
8739 and Pseudomonas aeruginosa (P. aeruginosa)
ATCC 15442. All types of test bacteria were obtained
from the Research Center for Pharmaceutical
Ingredients and Traditional Medicine, National
Research and Innovation Agency (BRIN), Kawasan
PUSPIPTEK, Serpong, South Tangerang, Banten,
Indonesia. Technical solvents were distilled before
maceration; isolation and spectral grade solvents ( nhexane, ethyl acetate (EtOAc), methanol (MeOH), and
dimethylene chloride (MTC) from Merck, Darmstadt,
Germany and Smart lab, Indonesia) were employed
for spectroscopic measurements.
Instrumentation
Optical rotations were measured on a Perkin Elmer
341 Polarimeter (Waltham, MA, USA). UV was
recorded on PerkinElmer UV WinLab Data Processor
and Viewer Version 1.00.00. The IR spectra were
recorded on a Perkin Elmer 1760 X FT-IR in KBr
(Waltham, MA, USA). Mass spectra were obtained with
a Waters Q-TOF. HR-MS XEVotm mass spectrometer
(Waters, Milford, MA, USA). NMR spectra of an epimer
mixture (2,3) was obtained with a Bruker Avance Neo
(BioSpin AG, Faellanden, Switzerland) at 700 MHz for
1
H and 175 MHz for 13C-NMR and NMR JEOL JNM
ECZ-600 (JEOL USA, INC.) spectrometer for 1, (at 600
MHz for 1H and 150 MHz for 13C-NMR), with CDCl3
as a solvent, chemical shift were given on a δ (ppm)
scale and both using tetra methyl silane (TMS) as the
internal standard.
Chromatographic separations
were carried out on silica gel 60 (Merck Kieselgel 60
PF 253 Art No. 7734.1000 and 9385.1000 with the
particle size 0.063-0.200 mm and 0.040 – 0.063
mm). Thin Layer Chromatography (TLC) plates were
precoated with silica gel GF254 (Merck, Darmstadt,
Germany, 0.25 mm). Spots were visualized under UV
light of 254 nm and 365 nm simultaneously and by
spraying with 10% Sulfuric acid (H2SO4) in ethanol or
vanillin reagent followed by heating.
Procedure

Extraction and isolation

The dried twigs (4.5 kg) of A. foveolata were
extracted with MeOH exhaustively (40L), at room
temperature for 5 days, 5 x 24 h. After removal of the

solvent under vacuum, the viscous concentrated
MeOH extract (233.5 g) was first suspended in water
(H2O) and then partitioned with n-hexane, EtOAc, and
n-butanol, successively. The EtOAc extract fraction
(46.3 g) was fractionated by vacuum liquid
chromatography (VLC) on silica gel 60 eluting
with a gradient of n-hexane:EtOAc: MeOH (100:0–
0:100, 2.5% v/v) to produce 110 fractions (1-110).
Fractions 9-11, white powder ( 6.68 g), were
chromatographed on a column of silica gel, eluted
with an isocratic eluent of n-hexane–EtOAc-CH2Cl2
(DCM) (9:0.5:0.5), to give 20 subfractions (9.01–
9.20). The combination of subfractions 9.05-9.10
with the major spot as the target was
chromatographed again on a silica gel column and
repeated with the same process and eluent until a pure
isolate of 1 (250) mg was obtained (Figure 1).
Fractions 29-30 (510 mg) were subjected to column
chromatography (CC) on silica gel (70–220 mesh)
eluted with n-hexane: EtOAc (7:3) to produce eight
subfractions (A-H) and the isolation process using CC
was repeated until the final isolation stage where it was
purified using non-polar silica gel, namely octa
dodecyl silica (ODS) and eluted with MeOH eluent to
produce a mixture of epimer compounds of 2 and 3
(12.5 mg), as shown in Figure 1.
Dammar-24-en-3β,20-diol (1). White amorphous
powder. Retention factor (Rf) of 1 on TLC = 0.8
(7:2:1= n-hexane: EtOAc: MTC), IR νmax (cm−1)
3343, 2921, 2852, 1743, 1465, 1376, 1070 cm-1.
0
[𝛼]29.6
𝐷 -0,10 (c, 0.26, CHCl3); No UV absorption was
detected. 1H-NMR (CDCl3, 600 MHz) and 13C-NMR
spectral data (CDCl3, 150 MHz) are shown in Table 1.
HR-TOFMS m/z found at 467.3863 [M+Na]+,
(calculated for C30H52O2Na = 467.3865).
Epimeric mixture of shoreic acid (2) and eichlerianic
acid (3). White amorphous powder. Rf of epimer 2 and
3 on TLC = 0.5 (7:3=n hexane: EtOAc), IR νmax
(cm−1) 3324, 2923, 2853, 1711, 1453, 1377, 1261,
0
1096, 1020.
[𝛼]28.7
0.26, CHCl3);
𝐷 +0,25 (c,
Ultraviolet (UV) spectra showed absorbance at 196
nm. 1H-NMR (CDCl3, 700 MHz) and 13C-NMR spectral
data (CDCl3, 175 MHz) are shown in Table 1. HRTOFMS m/z found at 497.3601 [M+Na]+ (calculated
for C30H50O4Na = 497.3607).

Determination of minimum inhibitory concentration
(MIC)

The MIC value of substances 1-3 was determined
using a standard microdilution assay with some
modifications (CLSI, 2022). A double dilution of the
sample solution (100 μl) was added to a 96-well sterile
microplate containing 100 μL Mueller-Hinton broth
medium (MHB). An antibacterial test was conducted
using a bacterial culture with titer 0.5 McFarland. For
that, the bacterial culture titer was adjusted so that the
OD value of 600 nm was 0.01 or equivalent to 1.5 x
107 CFU (Colony Forming Unit)/mL with the addition
of the required volume of 0.9% NaCl and smeared

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Molekul, Vol. 20. No. 2, July 2025: 394 – 403

Figure 1. Compounds 1- 3
into each well. This particular plate was further
incubated at 37 °C for 24 hours. The MIC values were
determined to be the lowest concentration of the
extract that could suppress the bacterial growth,
observed by the clear medium of the well and the
particular concentration with no bacterial growth
observed on the medium, respectively. Tetracycline
(Sigma-Aldrich) was used as the positive control.
RESULTS AND DISCUSSION
Three dammarane-type triterpenoids (Figure 1)
were obtained by separating and purifying the ethyl
acetate extract from the twigs of A. foveolata using the
column chromatography technique. Compound 1 was
obtained as a white powder. The molecular formula of
compound 1 was determined as C30H52O2 based on
the HR-TOFMS spectrum of m/z found at 467.3863
[M+Na]+, (calculated for C30H52O2Na = 467.3865)
with the NMR data (Table 1) and thus obtained five
degrees of unsaturation.
The UV spectrum of compound 1 showed no
conjugated double bonds based on the maximum
absorption at a wavelength above 200 nm. The IR
spectrum of compound 1 showed the presence of
hydroxyl groups (3343 cm-1), aliphatic C-H (2921 and
2852 cm-1), C=C (1743 cm-1), gem-dimethyl (1458
and 1376 cm-1), and C-O (1070 cm-1) functionalities.
The 1H-NMR (CDCl3, 600 MHz, ppm) spectrum of
compound 1) shows the presence of eight tertiary
methyl signals resonating at δH 0.96 (H-18), 0.86 (H19), 1.14 (H-21), 1.68 (H-26), 1.61 (H-27), 0.93 (H28), 0.84 (H-29), and 0.89 (H-30), one oxygenated
methine signal at δH 3.40 (1H, t, J = 4.5 Hz; H-3), and
one sp2 methyl proton signal resonates at δH 5.12

(1H, t, J = 5.5 Hz; H-24). The 13C-NMR (CDCl3, 150
MHz, ppm) spectrum of compound 1, detailed with
DEPT 135° experiments, shows the presence of 30
carbon signals consisting of eight methyl, ten
methylene, six methine, and six quaternary carbon
signals.
The presence of eight methyl signals resonating at
δC 15.5 (C-18), 16.0 (C-19), 25.4 (C-21), 25.8 (C26), 17.7 (C-27), 28.3 (C-28), 22.1 (C-29), and 16.5
(C-30) ppm, one oxymethine signal at δC 76.3 (C-3)
ppm, one oxygenated quaternary carbon signal at δC
75.4 (C-20) ppm, one sp2 methine signal at δC 124.7
(C-24) ppm, and one sp2 quaternary carbon signal at
δC 131.6 (C-25) ppm. This function is counted as one
of five degrees of unsaturation, the remaining four
degrees of unsaturation are derived from the resintype tetracyclic triterpenoid skeleton (Harneti et al.,
2023; Joycharat et al., 2010).
The determination of the location of the functional
groups from the structure of compound 1 was
confirmed by HMBC experiments. The correlations
between methyl protons CH3-21 (δH 1.14 ppm), and
H-22 (δH 1.45 ppm) with C-20 indicate the –OH group
is bound to C-20. Other –OH groups bound to C-3
are evidenced by the correlations between H-3 (δH
3.40) with C-1 (33.6) and C-5 (49.5), CH3-28 (δH
0.93), and CH3-29 (δH 0.84) with C-3 (76.3), C-4
(37.6), and C-5 (49.5). The correlations observed in
CH3-26 (δH 1.68), and CH3-27 (δH 1.61) with C-25
(131.6), and C-24 (124.7), and H-24 (δH 5.12) with
C-23 (22.5), C-26 (25.8), and C-27 (17.7) indicate
the presence of a double bond in the side chain of
compound 1, at the C-24/C-25 position as shown in
Figure 2.

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Dammarane-Type Triterpenoids

Ricson Pemimpin Hutagaol, et al.

Table 1. NMR data of compounds 1–3 in CDCl3 (δ in ppm, 175 MHz 13C-NMR, 700 MHz 1H-NMR for 2, 3 and
150 MHz, 600 MHz for 1)
1
2
3
No
δC
δH
δC
δH
δC
δH
(Integ., mult, J=Hz)
(Integ., mult, J=Hz)
(Integ., mult, J=Hz)
1
33.6
1.38 (1H, m)
34.3
1.61 (2H, m)
34.3
1.63 (2H, m)
1.42 (1H, m)
2
24.8
1.43 (1H, m)
28.2
2.18 (1H, m)
28.2
2.18 (1H, m)
1.74 (1H, m)
2.38 (1H, m)
2.38 (1H, m)
3
76.3
3.40 (1H, t, 4,5)
179.4
179.2
4
37.6
147.5
147.5
5
49.5
1.23 (1H, m)
50.9
1.97 (1H, m)
49.8
1.96 (1H, m)
6
18.2
1.39 (2H, m)
24.6
1.38 (2H, m)
24.6
1.36 (2H, m)
7
35.1
1.25 (1H, m)
33.9
33.9
1.55 (1H, m)
8
40.6
40.1
40.1
9
50.4
1.43 (1H, m)
41.2
1.47 (1H, m)
41.2
1.47 (1H, m)
10 37.2
39.1
39.1
11 21.4
1.51 (2H, m)
22.1
1.29 (2H, m)
22.3
1.27 (2H, m)
12 25.4
1.91 (1H, m)
27.2
26.9
1.52 (1H, m)
13 42.2
1.60 (1H, s)
43.0
42.9
14 50.4
50.4
50.4
15 31.1
1.05 (1H, m)
31.5
1.47 (2H, m)
31.5
1.48 (2H, m)
1.49 (1H, m)
16 27.5
1.78 (2H, m)
25.7
1.48 (2H, m)
25.8
1.48 (2H, m)
17 49.8
1.68 (1H, m)
49.5
1.88 (1H, m)
49.8
1.87 (1H, m)
18 15.5
0.96 (3H, s)
16.4
0.89 (3H, s)
16.3
0.89 (3H, s)
19 16.0
0.86 (3H, s)
20.2
0.85 (3H, s)
20.2
0.86 (3H, s)
20 75.4
86.4
86.5
21 25.4
1.14 (3H, s)
23.5
1.13 (3H, s)
27.1
1.13 (3H, s)
22 40.5
1.45 (1H, s)
35.8
1.67-1.81 (2H, m)
34.8
1.70-1.82 (2H, m)
23 22.5
2.05 (2H, m)
26,2
1.80 (2H, m)
26.4
24 124.7
5.12 (1H, t, 5,5)
83.3
3.73 (1H, t, 7.5)
86.6
3.62 (1H, t, 5.10)
25 131.6
71.6
70.3
26 25.8
1.68 (3H, s)
27.4
1.19 (3H, s)
27.8
1.15 (3H, s)
27 17.7
1.61 (3H, s)
24.3
1.12 (3H, s)
24.1
1.11 (3H, s)
28 28.3
0.93 (3H, s)
113.5
4.66 (1H, brs)
113.5
4.66 (1H, brs)
4.85 (1H, brs)
4.85 (1H, brs)
29 22.1
0.84 (3H, s)
23.2
1.73 (3H, s)
23.2
1.73 (3H, s)
30 16.5
0.89 (3H, s)
15.3
1.00 (3H, s)
15.4
1.02 (3H, s)
The 1H-1H COSY correlations of compound 1
indicates the presence of a basic framework of
dammarane
triterpenoid
compounds.
The
stereochemistry of compound 1 was determined
through a comparative study with the literature,
where the chemical shift (ppm) of carbon and
proton of compound 1 at C-3 [76.3; 3.40 (1H, J =
4.5Hz)], this indicates that the position of protons
2 and 3 are axially positioned, the hydroxyl group at
C-3 is equatorially positioned (3β) (Zhang et al.,
2010), and based on the biogenesis approach to the
existence of dammarane triterpenoids in the Genus
Aglaia. Thus, compound 1 was identified as 3β,20Sdihydroxy-dammar-24-en (Figure 1). Compound 1
has previously been isolated from the species Aglaia
elliptica (Meliaceae), where the H and C NMR data

obtained as a comparative reference can be seen in
Table 2 (Farabi et al., 2022).
Compounds 2 and 3 were obtained as an
inseparable mixture of epimers in a ratio of around
3:1, with 2 as a major compound. Epimeric
mixture was obtained as a white amorphous
powder, with the molecular formula C30H50O4. Its
molecular composition was established from the
HR-TOF MS found m/z 497.3601[M+Na]+
(calculated for C30H50O4Na, m/z 497.3607) and
NMR data (Table 1). The Hydrogen Deficiency (HD)
index was calculated using the equation HD = ∑C −
∑H/2 + 1, yielding an HD index of six for compounds
2 and 3 each. The IR spectra showed the presence of
a -OHst ( 3324 cm-1), a sp3-CH3 (2962 and 2923 cm1
), C=Ost (1711 cm-1), a gem-dimethyl (1457 and
1377 cm-1), C-Ost (1261, 1096, and 1020 cm-1).

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Table 2. NMR reference data for compounds 1–3 in CDCl3 (δ in ppm, 75 MHz 13C-NMR, 400 MHz
1
H-NMR for 2, 3, and 125 MHz, 500 MHz for 1)

No
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30

1 (3β,20S-dihydroxy2 (Shoreic acid,(Roux et
dammar-24-en,
al., 1998))
(Farabi et al., 2022))
δH
δH
δC
δC
(Integ., mult, J=Hz)
(J=Hz)
33.7
1.37 (1H, m)
34.0
1.40 (1H, m)
24.9
1.43 (2H, m)
28.1
2.18
2.38
76.4
3.37 (1H, t, J =
179.
4.5)
5
37.7
147.
2
49.6
1.23 (1H, m)
50.5
18.3
1.38 (2H, m)
24.3
35.2
1.24 (1H, m
33.6
1.55 (1H, m)
40.7
39.8
50.4
1.42 (1H, m)
40.9
37.3
38.8
21.4
1.52 (2H, m)
22.9
25.4
1.53 (1H, m)
27.0
1.91 (1H, m)
42.3
1,58 (1H, m)
42.7
50.5
50.1
31.2
1.04 (1H, m)
31.2
1.45 (1H, m)
27.6
1.77 (2H, m)
25.4
49.8
1.69 (1H, m)
49.2
15.6
0.93 (3H, s)
16.1
0.86 s
16.1
0.82 (3H, s)
19.9
0.82 s
75.5
86.1
25.5
1.13 (3H, s)
23.9
1.11 s
40.6
1.44 (2H, m)
35.5
22.6
2.02 (2H, m)
25.9
124.8
5.10 (1H, t, J =
83.1
3.71 dd (7.7)
5.4)
131.7
71.4
25.9
1.66 (3H, s)
27.1
1.18 s
17.8
1.59 (3H, s)
24.3
1.10 s
28.4
0.91 (3H, s)
113.
4.63 brs
2
4.82 brs
22.2
0.81 (3H, s)
23.2
1.70 s
16.6
0.86 (3H, s)
15.0
0.97 s

The 1H NMR (CDCl3, 700 MHz, ppm) spectrum of
2 displayed the presence of three tertiary methyl
groups at δH 0.89 (H-18), 0.85 (H-19), and 1.00 (H30), as well as three secondary methyl groups at δH
1,73 (H-29), 1.13 (H-21), 1.19 (H-26) and 1.12 (H27). The presence of seven methyl protons indicates
that compound 2 is a triterpenoid compound (Harneti
et al., 2012). The presence of a characteristic
oxygenated methine at δH 3.73 (1H, t, J = 7.5 Hz, H24) indicates the presence of a tetrahydrofuran ring in

3 (Eichlerianic acid, (Roux
et al., 1998))
δC

δH
(J=Hz)

34.1
28.1

2.18
2.38

179.5
147.1
49.7
24.4
33.7
39.9
41.0
38.9
22.2
26.7
42.8
50.2
32.0
25.6
49.6
16.1
20.0
86.4
27.0
34.6
26,2

0.87 s
0.84 s
1.13 s

86.2

3.62, dd (5.5, 10)

70.2
27.6
23.9

1.17 s
1.09 s

113.3

4.64 brs

23.1
15.2

4.83 brs
1.71 s
0.99 s

the side chain of a triterpenoid compound of the
dammarane group (Roux et al., 1998). In compound
2, a methylene sp2 signal was also observed at δH 4.66
and 4.85 ppm (1H, s, H-28).
The 13C NMR (CDCl3, 175 Hz, ppm) spectrum of 2,
showed 30 carbon resonances. These resonances
were classified by their chemical shifts, DEPT, and
HMQC spectra as follows: 7 methyl groups (three
tertiary at δC 16.4 (C-18), 20.2 (C-19), and 15.3
(C-30) ppm. 4 secondary at δC 23.2 (C-29), 23.5 (C-

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Ricson Pemimpin Hutagaol, et al.

21), 27.4 (C-26),
and 24.3 (C-27) ppm. 11
methylene groups of which one methylene olefinic,
five methine groups of which one is oxygenated
methine, seven quaternary carbons of which two
are oxygenated quaternary carbons at 86.4 (C-20)
and 71.6 (C-25)
ppm
which indicated the
presence of a typical tetrahydrofuran ring in the side
chain of dammarane-type triterpenoid (Roux et al.,
1998). In the spectrum, the signals of sp2 quaternary
carbon at δC 147.5 (C-4) and sp2 methylene at δC
113.5 (C-28) as well as carboxylic acid carbonyl at δC
179.4 (C-3) were also observed, indicating the
presence of a ring in the open dammarane
triterpenoid framework in compound 2 (Szoka et al.,
2024). The existence of these 2 functional groups
accounted for 2 out of the total 6 hydrogen deficiency
index. The remaining 4 hydrogen deficiencies were
consistent were consistent with the tetracyclic
triterpenoid structure of 2.
The functional group's position of 2 was deduced
from the HMBC spectra. The existence of open cyclic
A at positions 3 and 4 in compound 2 can be shown
by the absence of observed correlation between H-2
(δH 2.38) to C-4 or correlations between H-29 (δH
1.73) to C-3. In addition, the presence of a double
bond at C-4/C-28 can be seen by the correlations
between H-28 (δH 4.66 and 4.85 ppm) with C-4 and
C-5 as well as the correlations between H-29 (δH 1.73)
with C-4, C-28, and C-5. The presence of correlations
between the methyl protons H-21 (δH 1.15) with C-17,
C-22, and C-20, as well as correlations between H27 with C-24, C-26, and C-25, is also seen in the

spectrum of the correlations between H-26 (δH 1.21
ppm) with C-24, C-27, and C-25, which indicates the
presence of a tetrahydrofuran ring located at C-17
(Figure 1). The positions of the methyl groups seen
embedded at C-10, C-8, and C-14 are respectively
indicated by the correlations between H-19, H-18,
and H-30. The entire HMBC correlation of compound
2 can be seen as in Figure 2.
The 1H-1H COSY correlation of compound 2
indicates the presence of a basic framework of
dammarane triterpenoid compounds (Figure 2). The
presence of a cross peak between H1-H2 indicates an
open dammarane triterpenoid A ring, the cross peak
between H5-H6-H7 indicates an open dammarane
triterpenoid A ring at C-3/C-4. Thus, compound 2 has
a planar structure of 20,24-epoxy-25-hydroxy-3,4secodamar-4(28)-en-3-oic acid (shoreic acid) and its
complete correlations are shown in Figure 2.
To determine the stereochemistry of asymmetric
carbons it can be determined through the chemical
shift values of 1H and 13C-NMR. Compound 2 shows
chemical shifts for C-24 and H-24 respectively δC 83.3
and δH 3.73 ppm; dd, J = 4.8 and 10.8 Hz, and the
chemical shift of C-20 is 86.4 ppm, this indicates the
configuration of C-20 is S and C-24 is R (Roux et al.,
1998). Further support was obtained from the results
of the comparison of NMR data between compound 2
and shoreic acid in Tables 1 and 2. Based on the
spectral data obtained, previous research data, and
the approach to the biogenesis of dammarane
triterpenoid compounds, compound 2 was identified
as shoreic acid (Seger et al., 2008).
26

26
25

OH

21

20
11

12
18

19
9
2

1

3

4

HO
28

6

7

22

24

21

22

O
16

14

19

HO

9
2
29

30

1

12
18

11

3

15

29

OH

23

17

13

8

10
5

23

27

1
4

13

6

O
20
17
16

14
15

8

10
5

7

28

24

30

2

26

OH

23
24

22

25

27

O

21
20

O

19

12
18

11

13

9

3

HO

2
29

1
4

28

14

17
16
15

8

10
5
6

7

1H-1H COSY

30

HMBC

3

Figure 2. Selected HMBC and 1H-1H COSY correlations of 1 – 3

399

25

27

Molekul, Vol. 20. No. 2, July 2025: 394 – 403

Based on the reference search that has been done,
the mixed epimer compounds 2 and 3 have a structure
similar to the aglinin A compound that has been
successfully isolated from several species, such as A.
forbesii, A. smithii was also found in the form of a
mixture of its epimers, 24R and 24S. The
tetrahydrofuran ring with hemiketal in molecule 2
might be interconvertible with an intermediate 20-ol24-one form (through a ring opening and closing
procedure under protic conditions, which results in the
epimer mixture 2 and 3 being inseparable (Feng et al.,
2013).
The 1D NMR chemical shift and spectrum of 3 in
the epimeric mixture as can be seen in Table 1, shows
duplication in almost all signals but with the signal of
compound 2 being more dominant than compound 3
with a ratio of approximately 3:1, that was indicating
the presence of two epimeric compounds. The 13C
NMR spectrum of compounds 3 and 2, as shown in
Table 1 shows duplication in 29 signals from 30
carbon signals, which indicates the presence of two
epimeric compounds. In the 1D NMR spectrum only
one significant difference is seen at the C-24 position,
where compound 2 shows a chemical shift for C-24
and H-24 respectively δC 83.3 ppm and δH 3.73 ppm
(1H, t, J = 7.5 Hz), while the chemical shift at C-20 is
86.4 ppm, this indicates that the configuration of C20 is S and C-24 is R for compound 2.
Meanwhile, compound 3 shows the chemical shift
δC 86.6 ppm and δH 3.62 ppm (1H, t, J = 5.10 Hz),
while the chemical shift at C-20 is 86.5 ppm, this
indicates that the configuration of C-20 is S and C-24
is S for compound 3, it can be seen that 2 and 3 is an
epimeric isomer compound at C-24. Shoreic acid (2),
whose stereochemistry has been determined by X-ray
(Lavie et al., 1984), is the 24R isomer, while 3 is the
24S isomer. Experiments with 2D NMR allow to obtain
the correlations and structure of the two compounds
as shown in Figures 1 and 2 and also the exact 13C
and 1H chemical shift data of 2 and 3 as shown in
Table 1, and the comparative NMR data can be seen
as in Table 2. Based on the spectral data obtained,
which is in accordance with the literature, the name of
compound 3 was confirmed to be eichlerianic acid.
The antibacterial activities of the isolated
compounds were evaluated against normal ATCC
bacteria strains (Tables 3). To assess the potential of
antimicrobial compounds in this study, the broth
microdilution method was employed, which yields
quantitative
data,
specifically the MIC value
(Swebocki et al., 2023). MIC is the lowest
concentration of an antimicrobial agent that can
inhibit the growth of certain bacteria, which can be
observed with the naked eye through changes in
turbidity (Hossain, 2024). Turbidity values are
measured by inserting a microplate into a plate
reader and reading the OD600 value from the wells.
Then, the data is collected and the MIC value is
calculated (Kadeřábková et al., 2024).

The epimeric mixture of 2 and 3 showed moderate
antibacterial activity with a minimum inhibitory
concentration value ranging from 31.7 to 126.6 ppm,
particularly against S. aureus with a MIC value of 31.7
ppm, while compound 1 was inactive against all types
of bacteria. Based on the activity data of the
triterpenoid compounds, it is suspected that the
functional groups, namely (-COOH) and the double
bond in the open cyclic A found in the mixed
compounds of epimers 2 and 3, play a role in
increasing their activity.
The epimers 2 and 3 are more active against
gram-positive bacteria, particularly S. aureus, than
gram-negative bacteria. This is also thought to be
because gram-negative bacteria have a doublelayered membrane structure, a lipopolysaccharide
layer, membrane proteins, and porins that are not
found in gram-positive bacteria, thus inhibiting the
activity of compounds (Hickson et al., 2025; Ji et al.,
2022; Woods et al., 2025). S. aureus is a Grampositive species that belongs to the family
Micrococcaceae. It is commonly found on human skin
and in the nose (Rajput et al., 2024).
Based on this research, these epimer compounds
have the potential to be further researched and
developed as an antibiotic compound against S.
aureus. The electron-withdrawing group (-COOH)
bound to the open cyclic A is thought to contribute
to
the
increased
antibacterial activity
(Purwantiningsih et al., 2020). Carboxylic organic
acids have antimicrobial potential due to the
lipophilic nature of the undissociated acid form
(RCOOH) which can penetrate the microbial plasma
membrane only through passive diffusion (Mira et
al., 2024). Organic acids seem to have varied
putative antibacterial mechanisms involved in
cytoplasmic acidification, osmotic stress, membrane
disintegration,
and
cytoplasmic
enzyme
destabilization (Yoon et al., 2024). The antibiotic
tetracycline, used as a positive control, showed strong
activity against all four bacterial species, with an MIC
value of 1.55 ppm. Tetracycline inhibits protein
biosynthesis by targeting ribosomal subunit 305.
Tetracycline inhibits the binding of tRNA to the
ribosomal
A
site, ultimately inhibiting protein
synthesis (Halawa et al., 2023).
Based on several references that have been
successfully
obtained,
it is known that the
antibacterial activity of triterpenoid compounds is
influenced by the presence and position of functional
groups such as carboxylic acids and double bonds
contained in the compound. Two triterpenoid
compounds
without
carboxylic acid functional
groups,
lanosterol
and
lupeol, have been
successfully isolated from Euphorbia arbuscula and
tested for their antibacterial activity against S.
aureus, E. coli, and P. aeruginosa. The isolated
compounds showed weak or no antibacterial activity.
(Al-Ansi et al., 2024).

400

Dammarane-Type Triterpenoids

Ricson Pemimpin Hutagaol, et al.

Table 3. MIC values of the isolated compound against ATCC strains.
Compound
1
Epimeric mixture of
2 and 3
Tetracycline
(Control)

Bacterial tested

P. aeruginosa

E. coli

> 225

MIC (ppm)
> 225
> 225

> 225

126.6

63.3

31.7

63.3

1.55

1.55

1.55

1.55

Three tetracyclic triterpenoic acid compounds
having carboxylic acid groups in their structure,
namely densiflorinic acid A-C, have been successfully
isolated from Dysoxylum densiflorum and tested for
their antibacterial activity. These compounds were
tested against seven bacterial species using the
microdilution method. They exhibited only weak
antibacterial properties, with densiflorinic A exhibiting
the highest activity against B. subtilis, with an MIC of
26.5 μM (Komang et al., 2016).
CONCLUSIONS
The ethyl acetate preparation of A. foveolata twigs
produced three dammarane-type triterpenoids and
one steroid, which were identified as dammar-24-en3β,20-diol (1), an epimeric mixture of shoreic acid (2);
eichlerianic acid (3). The antibacterial activities of the
isolated compounds were evaluated against four
normal ATCC bacteria strains, including two grampositive bacteria (S. aureus, and B. subtilis) and two
gram-negative bacteria (E. coli, and P. aeruginosa).
An epimeric acid mixture of shoreic (2) and
eichlerianic acid (3) was highly active (MIC 31.7 ppm)
against S.aureus and active to inhibit other normal
bacterial strains.
ACKNOWLEDGEMENTS
The author would like to thank the National
Research and Innovation Agency (BRIN) through the
author's ELSA for providing access to the NMR data
used in this study.
CONFLICT OF INTEREST
No potential conflict of interest was reported by the
authors.
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