melaleuca ericifolia sm
TRANSCRIPT
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
110
RADICAL SCAVENGING POTENTIAL OF COMPOUNDS FROM
MELALEUCA ERICIFOLIA SM.
Fatma M. Abdel Bar1*
,2
1Department of Pharmacognosy, Faculty of Pharmacy, Prince Sattam Bin Abdulaziz
University, Al-Kharj 11942, Saudi Arabia.
2Department of Pharmacognosy, Faculty of Pharmacy, Mansoura University, Mansoura
35516, Egypt.
ABSTRACT
Plants belonging to genus Melaleuca are economically important
plants as the leaves of most species contain volatile oils. In this study,
the non-volatile constituent of the leaf and bark of Melaleuca ericifolia
Sm. grown in Egypt was investigated. The leaf extract afforded (E)-
3`,4`-dimethoxy cinnamaldehyde 1, methyl tri-O-methyl gallate 2,
kaempferol 3, 3,3`,4`-trimethoxy ellagic acid-4-O--glucoside 4 and 3-
methoxyellagic acid 4-O--rhamnopyranoside 5. While the bark
extract afforded melaleucic acid 6 and gallic acid 7. The structures of
the isolated compounds were identified using 1D and 2D NMR
spectroscopy. The antioxidant activities represented by %inhibition
and free radical scavenging profile of ABTS+ radical was evaluated at
25, 50, 100 and 200 M, for 30 min. The results showed that gallic
acid 7 (88% inhibition, at 25 M) and kaempferol 3 (85% inhibition, at 100 M)
respectively, were the most active compounds with comparable activity to ascorbic acid as a
positive control. Remarkably, the lupane-type triterpene melaleucic acid 6 showed
considerable ABTS+ radical bleaching activity (53.7 %inhibition at 200 M, after 5 min). In
conclusion, this study reported for the first time the isolation of compounds 4 and 6 from this
plant and the antioxidant activity of melaleucic acid.
KEYWORDS: Melaleuca ericifolia; melaleucic acid; 3,3,4`-trimethoxy ellagic acid-4-O-
glucoside; antioxidant activity; radical scavenging activity.
World Journal of Pharmaceutical Research SJIF Impact Factor 8.074
Volume 8, Issue 9, 110-123. Research Article ISSN 2277– 7105
Article Received on
04 June 2019,
Revised on 25 June 2019,
Accepted on 15 July 2019,
DOI: 10.20959/wjpr20199-15478
*Corresponding Author
Dr. Fatma M. Abdel Bar
Department of
Pharmacognosy, Faculty of
Pharmacy, Prince Sattam
Bin Abdulaziz University,
Al-Kharj 11942, Saudi
Arabia.
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
111
INTRODUCTION
Melaleucas are commonly known "paper-bark trees". The essential oil of Melaleuca
alternifolia, ―tea tree oil‖ is the most common one. Melaleuca ericifolia Sm. is an evergreen
bushy shrub to small tree, with grayish papery bark, needle-like leaves and white to cream
scented bottlebrush-like flowers, known as the Swamp Paperbark tree.[1]
Phytochemical
studies on M. ericifolia Sm. is mainly concerned with the volatile oil of this plant. Great
variation in the chemical composition of the volatile oil has been observed based on its
geographical source. The chemical composition of essential oil of M. ericifolia Sm. growing
in southeastern Australia was composed mainly from the terpenoids, linalool 60% v/v and
1,8-cineole 16%v/v.[2]
However, the essential oil of M. ericifolia Sm. grown in Egypt was
shown to contain the phenylpropanoid, methyleugenol 96.84% v/v.[3,4]
The essential oil of M.
ericifolia Sm. showed antibacterial, antifungal, antiviral activities (Farag et al. 2004, Farag
1998). Additionally, the antioxidant activity of the essential oil of M. ericifolia Sm. has been
investigated by the same work group.[4]
The presence of pentacyclic triterpenes including 28-norlupane derivatives, betulinic acid,
betulinaldehyde, betulin, platanic acid, ursolaldehyde, ursolic acid, and asiatic acid has been
previously reported.[5]
The antiprolifrative activity of these pentacyclic triterpenes have been
also described.[5]
Regarding the phenolic contents of this plant, several phenolics have been
identified including ericifolin, gallic and ellagic acid derivatives, in addition to flavonoids
such as kaempferol, quercetin, myricetin and their glycosides.[6]
The hepatoprotective effect
of praecoxin A, a polyphenolic from M. ericifolia Sm., has been also investigated against
CCl4‐induced toxicity in mice.[7]
Nevertheless, no studies addressed the antioxidant activity of the non-volatile compounds
isolated from this plant. Therefore, this work aimed at isolation and characterization of the
non-volatile constituents of the leaf and bark extracts of M. ericifolia Sm. and investigation of
their antioxidant activity using ABTS+ radical bleaching assay.
MATERIALS AND METHODS
Plant Materials
Branches of M. ericifolia Sm. growing in Mansoura University, Egypt, were collected in
January 2004, leaves and barks were separated, dried and powdered. The authentication of
the plant was kindly implemented by Prof. Dr. Sayed F. Khalifa, Faculty of Sciences, Ain
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
112
Shams University, Cairo, Egypt. A voucher specimen was deposited at the Pharmacognosy
Dept., Mansoura University, Egypt (06-Mansoura-1).
General experimental
The 1H and
13C NMR spectra were recorded in CDCl3, CD3OD, pyridine-d6 and DMSO-d6
(dimethyl sulfoxide) using TMS as an internal standard, on a JEOL Eclipse NMR
spectrometer operating at 400 MHz for 1H and 100 MHz for
13C. The solvents used for
extraction and chromatographic separations were of reagent grade. TLC analysis was carried
out on precoated silica gel 60 F254 500 µm TLC plates, using the developing systems
chloroform-methanol (9:1). For TLC, silica gel 60 F254 plates (EMD Chemicals Inc.) were
used. For CC., silica gel 60 (EMD Chemicals Inc.), 70-230 mesh, or Silica gel (Natland
International Corporation), 230-400 mesh, were used. For gel permeation CC. Sephadex LH20
(SIGMA-ALDRICH) was used. For ABTS+
radical assay, Azino-bis-(3-ethylbenzthiazoline-
6-sulfonic acid) was purchased from Fluka (Buchs, Switzerland); MnO2 was purchased from
Sigma–Aldrich (Steinheim, Germany), Ascorbic acid (Memphis Pharmaceutical Co., Cairo,
Egypt), Phosphate buffer from BDH Chemicals (London, UK) and absorbance was obtained
using a Bio-Tek Elisa microplate reader (Winooski, VT, USA).
I- Phytochemical investigation of leaves of M. ericifolia Sm.
Extraction
The powdered leaves of Melaleuca ericifolia Sm. (2.5 kg) were extracted with methanol (7 x
2 L) and methanol-water (7.5:2.5, 2 x 2 L). The combined methanol extract was then
concentrated under vacuum at 45 C°. The dry methanol extract (915 g) was dissolved in
methanol-water (1.5: 0.5 L). The hydro-alcoholic solution was partitioned successively using
n-hexane, chloroform and EtOAc. The obtained extracts were evaporated under vacuum at
45°C and kept for phytochemical investigation.
Column chromatographic investigation of n-hexane extract of M. ericifolia Sm. leaves
The n-hexane extract (135 g) was chromatographed over silica gel CC. and eluted with n-
hexane-EtOAc mixtures of increasing polarity starting with n-hexane 100%. Fractions of 200
mL were collected to give 50 fractions. Similar fractions were then grouped and purified as
follows.
Fraction (12-16), 15.6 g, eluted with n-hexane-EtOAc (9.5: 0.5), revealed the presence of
one major spot [Rf 0.21, n-hexane-EtOAc (8:2)]. Purification using silica gel CC. and n-
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
113
hexane-EtOAc (gradient) give 91 sub-fractions. Sub-fraction (77-88), 70 mg, was further
purified using medium-pressure liquid chromatography (MPLC), C18 silica gel and water-
methanol (50:50) to afford compound 1, 12 mg.
Fraction 5 (17-28), 5 g, eluted with n-hexane-EtOAc (9.5:0.5), revealed the presence of
presence of seven major spots [n-hexane-EtOAc (8:2)]. This fraction was purified by
repeated column chromatography using silica gel and n-hexane-EtOAc (9.5:0.5), 50 mL
fractions were collected to give 37 sub-fractions. Sub-fraction (22-30), 2.5 g, was further
fractionated using silica gel CC., n-hexane-EtOAc (gradient elution) and fractions of 50 mL
were collected to give 59 sub-fractions. Sub-fraction (34-41), 1.3 g, was further purified
using MPLC, C18 silica gel and water-methanol, gradient elution. Fractions of 5 mL were
collected to obtain sub-fraction (21-26), eluted with water-methanol (3:7), which afforded
compound 2 [Rf 0.37, n-hexane-EtOAc (8:2)], 200 mg.
Investigation of the chloroform extract of M. ericifolia Sm. leaves
The chloroform extract was divided based on its components into two parts (A and B).
Chloroform-A extract was obtained as the first crop and contained a large amount of gel
precipitate that was evaporated into a yellowish solid (142.0 g, 5.6 %w/w). Unlike the
chloroform-B which was obtained as a brown viscous liquid (14.0 g, 0.6 %w/w).
The chloroform-B extract, 14 g, was fractionated using silica gel and chloroform-methanol,
gradient elution and fractions of 50 mL were collected to give 48 fractions purified as
follows.
Fraction (21-23), 300 mg, eluted with chloroform-methanol (9.5: 0.5), revealed the presence
of two major spots. The fraction was further purified by gel permeation on Sephadex LH20
column and chloroform-methanol, gradient elution and fractions of 5 mL were collected to
obtain 420 sub-fractions. Sub-fractions (370-400), eluted with chloroform-methanol (8:2)
afforded compound 3 [Rf 0.77, chloroform-methanol (8:2)], 15 mg.
Fraction (31-33), eluted with eluted with chloroform-methanol (9:1), afforded compound 4
[Rf 0.23, chloroform-methanol (9:1)], 100 mg.
Fraction (39-41), eluted with chloroform-methanol (8:2), afforded compound 5 [Rf 0.13,
chloroform-methanol (9:1)], 35 mg.
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
114
II- Phytochemical investigation of the bark of M. ericifolia Sm.
Extraction
The powdered bark of Melaleuca ericifolia (900 g) was extracted with methanol (5 x 1 L) and
methanol-water (7.5:2.5, 2 x 1 L). The combined methanol extract was concentrated under
vacuum at 45 °C. The dry methanol extract (90 g), was partitioned using n-hexane and
EtOAc, respectively.
Column chromatographic investigation of the EtOAc extract of the bark
The EtOAc extract (12 g) was purified using gel permeation on Sephadex LH20 column,
chloroform-methanol (gradient elution) and sub-fractions of 50 mL were collected to give 82
fractions. Similar fractions were then grouped and purified as follows:
Fraction (38-39), 1 g, eluted with chloroform-methanol (9:1), revealed the presence of one
major spot [Rf 0.51, chloroform-methanol (9.5:0.5)]. The fraction was further purified using
silica gel and gradient elution with chloroform-methanol. Fractions of 25 mL were collected.
Sub-fraction (8-12), eluted with chloroform-methanol (9.8:0.2) afforded compound 6, 10 mg.
Fraction (75-82), 200 mg, eluted with chloroform-methanol (8:2), revealed the presence of
one major spot [Rf 0.76, EtOAc-methanol (8:2)]. The fraction was further purified using
MPLC, C18 silica gel and water-methanol (gradient elution). Fractions of 5 mL were
collected. Sub-fractions (14-30) eluted with water-methanol (9:1) afforded compound 7, 25
mg.
Antioxidant ABTS+ radical bleaching assay
The procedure was according to the published methods with some modification.[8-11]
Thoroughly, for preparation of the derived ABTS+ radical, 2 mL of ABTS solution (60 mM)
was added to 3 mL of MnO2 solution (25 mg/mL), all prepared in aqueous phosphate buffer
solution (pH 7, 0.1 M). The mixture was shaken, centrifuged and decanted. The formed
radical cations are almost prompt and stable for several hours. Duplicates were prepared for
each sample in a 96-well microplate. The absorbance (A) of the preformed ABTS+
radical
blue-green solution at 734 nm was adjusted to approximately 0.5 (Acontrol). The absorbance
(Atest) was measured upon the addition of aliquots of 20 L of the test sample (200, 100, 50,
25 M) in spectroscopic grade MeOH–buffer (1:1 v/v) to 180 L the ABTS•+
solution.
Duplicate of each sample was used in this assay. Ascorbic acid was used as a positive control.
Blank sample was run without ABTS+ radical solution and using MeOH–phosphate buffer
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
115
(1:1) instead samples. Negative control was run with ABTS•+
and MeOH–phosphate buffer
(1:1) only.
The %inhibition was calculated from the equation:
%Inhibition = Acontrol – Atest/ Acontrol x 100
RESULTS AND DISCUSSION
Identification of the isolated compounds
The structures of the isolated compounds (1-7), Figure 1, were identified using 1D and 2D
NMR spectroscopy. Based on 1H and
13C NMR data of compound 1 was identified as the
known compound (E)-3`,4`-dimethoxy cinnamaldehyde. Compound 1 has not been isolated
before from genus Melaleuca.
Compound 2 showed a good example of symmetry in 1H and
13C NMR data. Compound 2
was assigned based on its NMR spectra and comparison with published data[12]
to be methyl
tri-O-methyl gallate, which has not been isolated before from genus Melaleuca.
The structure of 3 was determined based on 1H and
13C NMR to be the flavonol aglycone,
kaempferol which has been isolated before from M. ericifolia Sm.[6]
Assignment of 1H and
13C NMR data of 4, HETCOR and HMBC determined its structure to
be the known compound, 3,3`,4`-trimethoxy ellagic acid-4-O- -glucoside.[13]
It is worth to
note that this is the first report of this compound in genus Melaleuca.
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
116
Figure 1: Structures of the isolated compounds from Melaleuca ericifolia.
Compound 5 was determined based on 1H and
13C NMR data (Table 1) to be 3-
methoxyellagic acid 4-O--rhamnopyranoside which has been isolated before from M.
ericifolia Sm. (Hussein et al., 2007). However, the underlined 13
C NMR C values (Table 1)
are miss-assigned as C-3 replaced C-4 and C-6 replaced C-6`. The true values are corrected
based on HMBC correlations (Figure 2a) that showed clearly 3J-HMBC correlation of the
methoxyl group (C-8) with a quaternary signal at 142.2 (C-3) and 3J-HMBC correlation of
the anomeric proton H-1`` with a quaternary carbon signal at 146.9 (C-4). Similarly, the C
values for C-6 and C-6` are corrected based on 2J-HMBC correlation of the proton singlet at
7.69 (H-5) with a quaternary carbon at 107.4 (C-6) and the proton singlet at 7.48 (H-5`)
with the carbon signal at 111.7 (C-6`), Figure 2b.
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
117
(a)
(b)
Figure 2a,b. Expanded HMBC correlations of compound 5.
Table 1: 1
H and 13
C NMR Data of Compound 5 (DMSO-d6, at 400 MHz for 1H
and 100
MHz for 13
C, Coupling constants (J) are in Hz).
Position H (J=Hz) C (ppm) HMBC
1 --- 114.7, qC
2 --- 140.6, qC
3 --- 142.2, qC
4 --- 146.9, qC
5 7.69, s 112.0, CH 1, 3, 4, 6, 7
6 --- 107.4, qC
7 --- 159.2, qC
8-OCH3 4.03, s 61.3, CH3 3
1` --- 113.5, qC
2` --- 136.5, qC
3` --- 140.6, qC
4` --- 153.1, qC
5` 7.48, 5 111.9, CH 1`, 3`, 4`, 6`, 7`
6` --- 111.7, qC
7` --- 159.2, qC
1`` 5.47, s 100.6, CH 4, 2``, 3``, 5``
2`` 4.01, m 70.5, CH 4``
3`` 3.86, m 70.6, CH 5``, 4``
4`` 3.33, m 72.3, CH
5`` 3.52, m 70.4, CH
6``(Me) 1.13, d (6.2) 18.4, CH3
1H NMR of compound 6 (Table 2) showed only 5 methyl signals and two exomethylene
protons at 4.82 (Ha -29) and 5.06 (Hb -29) suggesting a lup-20-ene nucleus with the
oxidation of two methyl into either CHO and/or COOH. 13
C NMR data showed two
quaternary carbon signals at 178.4 and 179.2 assigned to the two carboxylic groups at C-
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
118
27 and C-28, respectively. Different protons were assigned using HETCOR and COSY
correlations.
Table 2: 1H and
13C NMR data of compounds 6 (In pyridine-d6 at 400 MHz for
1H and
100 MHz for 13
C, Coupling constants (J) are in Hz).
Position H (J=Hz) C (ppm)
1 1.18, m , 1.80, m 39.3, CH2
2 1.76, m, 1.95, m 28.2, CH2
3 3.20, dd (11.0, 5.1) 77.7, CH
4 --- 39.1, s
5 1.00, m 55.9, CH
6 1.62, m, 1.80, m 18.7, CH2
7 1.96, m, 2.88, m 35.4, CH2
8 --- 40.7, s
9 2.25, m 52.3, CH
10 --- 37.7, s
11 1.34, m, 1.51, m 21.1, CH2
12 2.70, m, 2.09, m 26.9, CH2
13 2.93, m 40.2, CH
14 --- 59.9, s
15 2.54, m, 1.94, m 28.6, CH2
16 1.90, m, 2.06, m 38.3, CH2
17 --- 56.4, s
18 2.14, dd (11.4, 11.4) 51.6, CH
19 3.68, ddd (10.6, 10.6, 4.0) 47.8, CH
20 --- 151.0, s
21 1.56m, 2.54m 31.0, CH2
22 1.53m, 2.27m 37.6, CH2
23 1.04, s 28.4, q
24 0.97, s 16.4, q
25 0.86, s 16.9, q
26 1.18, s 17.4, q
27 --- 178.4, s
28 --- 179.2, s
29 4.82, brs, 5.06, brs 110.2, CH2
30 1.92, s 19.1, q
The structure of 6 was confirmed based on different NMR spectral method to be the known
compound melaleucic acid which has been isolated before from the bark and heartwood of
some species from genus Melaleuca.[14-16]
However, this is the first report of this compound
in Melaleuca ericifolia.
Based on 1H and
13C NMR, compound 7 was assigned to be gallic acid which has been
isolated before from different Melaleuca species.[17-19]
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
119
Antioxidant activity
Evaluation of the antioxidant activity of the isolated compounds (1-7) using ABTS+ radical
bleaching assay showed that kaempferol (3) and gallic acid (7) were the most active among
the tested compounds (Figure 3).
Figure 3: Antioxidant activity of the different compounds (1–7) using ABTS+ radical
scavenging assay and ascorbic acid (As.) as a positive control after 30 min, at different
concentrations (200, 100, 50, 25 M), absorbance was measured at 750 nm.
Gallic acid (7) showed potent antioxidant activity (85-88 %inhibition) comparable to the
positive control, ascorbic acid at all tested concentrations (25, 50, 100 and 200 M), Figure 3,
4a, 4d. This may be explained by the presence of three adjacent free phenolic OH groups.[20]
Likewise, kaempferol (3) showed strong antioxidant activity at 50, 100 and 200M (77-85
%inhibition), Figure 3, 4b, due to two meta phenolic OH in ring-A.[20]
Interestingly, the
lupane-type triterpene melaleucic acid (6) showed considerable ABTS+ radical bleaching
activity started after 5 min and become constant during the experiment time (53.7% inhibition
at 200 M), Figure 3, 4c. Thus, this study reported for the first time the antioxidant activity of
melaleucic acid. For the ellagic acid derivatives (4 and 5), compound (5) showed slight
antioxidant activity (5-30 %inhibition) due to the presence of only two phenolic OH groups
in the bulky structure. However, compound 4 did not show any significant ABTS+ radical
bleaching activity. This is can be explained by the lack of any free phenolic OH groups in the
structure of 4.[20]
Similarly, compounds 1 and 2 showed insignificant activity for the same
reason.
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
120
Figure 4: Time suppression profile of free radical scavenging activity of ABTS+ radical
at different concentrations (750 nm); (a) compound 7; (b) 3; (c) 6; (d) ascorbic acid.
(E)-3`,4`-dimethoxy cinnamaldehyde 1, NMR data (In CDCl3, at 400 MHz for 1H and 100
MHz for 13
C, coupling constants (J) are in Hz, carbon multiplicities were determined by APT
experiment); H 9.65, d (8.0 Hz, H-1), 6.58, dd (15.7, 8.0 Hz, H-2), 7.40, d (15.7 Hz, H-3),
7.07, d (1.8 Hz, H-2`), 6.89, d (8.4 Hz, H-5`), 7.15, dd (8.4, 1.8 Hz, H-6`), 3.92, s (H3-7`),
3.93, s (H3-8`).13
C NMR data;C 193.8, CH (C-1), 126.8, CH (C-2), 153.0, CH (C-3), 127.1,
qC (C-1`), 109.8, CH (C-2`), 152.9, qC (C-3`), 149.7, qC (C-4`), 111.1, CH (C-5`), 123.6,
CH (C-6`), 56.1, CH3 (C-7`), 56.0, CH3 (C-8`).
Methyl tri-O-methyl gallate 2, NMR data (In CDOD3, at 400 MHz for 1H and 100 MHz for
13C, coupling constants (J) are in Hz, carbon multiplicities were determined by APT
experiment);H 7.27, s (H-3/7), 3.82, s (H3-1`/3`), 3.87, s (H3-2`), 3.80, s (H3-4`).13
C NMR
data; C 166.7, qC (C-1), 125.2, qC (C-2), 106.5, CH (C-3), 153.0, qC (C-4), 142.1, qC (C-5),
153.0, qC (C-6), 106.5, CH (C-7), 55.3, CH3 (C-1`/3`), 59.8, CH3 (C-2`), 52.4, CH3 (C-4`).
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
121
Kaempferol 3, NMR data (In DMSO-d6, at 400 MHz for 1H and 100 MHz for
13C, coupling
constants (J) are in Hz, carbon multiplicities were determined by APT experiment); H 6.19,
d (1.8 Hz, H-7), 6.44, d (1.8 Hz, H-9), 8.03, d (8.8 Hz, H-2`), 6.92, d (8.8 Hz, H-3`), 6.92, d
(8.8 Hz, H-5`), 8.03, d (8.8 Hz, H-6`).13
C NMR data; C 147.3, qC (C-2), 136.2, qC (C-3),
176.5, qC (C-4), 161.3, qC (C-5), 98.8, CH (C-6), 164.5, qC (C-7), 94.0, CH (C-8), 159.8, qC
(C-9), 103.6, qC (C-10), 122.2, qC (C-1`), 130.1, CH (C-2`/6`), 116.0, CH (C-3`/5`), 156.7,
qC (C-4`).
3,3`,4`-Trimethoxy ellagic acid-4-O--glucoside 4, NMR data (In DMSO-d6, at 400 MHz for
1H and 100 MHz for
13C, coupling constants (J) are in Hz, carbon multiplicities were
determined by APT experiment); H 7.58, s (H-5), 4.04, s (8-OCH3), 7.82, s (H-5`), 4.11, s
(8`-OCH3), 3.99, s (9`-OCH3), 5.16, d (7.4 Hz, H-1``), 3.34, m (H-2``), 3.37, m (H-3``), 3.21,
m (H-4``), 3.43, m (H-5``), 3.50, m (H-6``a), 3.70, m (H-6``b). 13
C NMR data; C112.8, qC
(C-1), 141.7, qC (C-2), 141.4, qC (C-3), 152.5, qC (C-4), 108.0, CH (C-5), 113.3, qC (C-6),
158.9, qC (C-7), 61.9, CH3 (8-OCH3), 113.1, qC (C-`), 141.7, qC (C-2`), 142.2, qC (C-3`),
154.8, qC (C-4`), 112.5, CH (C-5`), 114.1, qC (C-6`), 158.7, qC (C-7`), 62.2, CH3 (8`-
OCH3), 57.3, CH3 (9`-OCH3), 101.8, CH (C-1``), 73.9, CH (C-2``), 77.0, CH (C-3``), 70.0,
CH (C-4``), 77.8, CH (C-5``), 61.1, CH2 (C-6``).
CONCLUSIONS
In this research, the chemical composition of the Swamp Paperbark tree, Melaleuca ericifolia
Sm. has been investigated. The isolation and characterization of seven known compounds
were reported from the leaves and barks of this plant. Compounds 4 (3,3`,4`-trimethoxy
ellagic acid-4-O--glucoside) and compound 6 (melaleucic acid) were reported for the first
time from this plant. The results of ABTS+ radical scavenging assay showed that gallic acid 7
and kaempferol 3 were the most active compounds while, the lupane-type triterpene
melaleucic acid 6 showed moderate antioxidant activity.
REFERENCES
1. Hewitt A. Melaleucas: Their Botany, Essential Oils and Uses by Joseph J. Brophy,
Lyndley A. Craven and John C. Doran. Australian Centre for International Agricultural
Research, ACIAR Monograph No. 156, Canberra, 2013, 415 pp. Price A$85.00
(paperback). ISBN 978-1-922137-51-7 (also available online ISBN 978-1-922137-52-4).
Austral Ecology, 2015; 40(8): e12-e13.
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
122
2. Brophy JJ, Doran JC. Geographic Variation in Oil Characteristics in Melaleuca ericifolia.
Journal of Essential Oil Research, 2004; 16(1): 4-8.
3. Abdel Bar FM, Khanfar MA, Elnagar AY, et al. Design and pharmacophore modeling of
biaryl methyl eugenol analogs as breast cancer invasion inhibitors. Bioorganic &
Medicinal Chemistry, 2010; 18(2): 496-507.
4. Farag RS, Shalaby AS, El-Baroty GA, Ibrahim NA, Ali MA, Hassan EM. Chemical and
biological evaluation of the essential oils of different Melaleuca species. Phytotherapy
research: PTR, 2004; 18(1): 30-35.
5. Abdel Bar FM, Zaghloul AM, Bachawal SV, Sylvester PW, Ahmad KF, El Sayed KA.
Antiproliferative triterpenes from Melaleuca ericifolia. J Nat Prod, 2008; 71(10):
1787-1790.
6. Hussein SA, Hashim AN, El-Sharawy RT, et al. Ericifolin: an eugenol 5-O-
galloylglucoside and other phenolics from Melaleuca ericifolia. Phytochemistry, 2007;
68(10): 1464-1470.
7. Al-Sayed E, Abdel-Daim MM, Khattab MA. Hepatoprotective activity of praecoxin A
isolated from Melaleuca ericifolia against carbon tetrachloride-induced hepatotoxicity in
mice. Impact on oxidative stress, inflammation, and apoptosis. Phytotherapy research:
PTR., 2019; 33(2): 461-470.
8. Lissi EA, Modak B, Torres R, Escobar J, Urzua A. Total antioxidant potential of resinous
exudates from Heliotropium species, and a comparison of the ABTS and DPPH methods.
Free Radic Res, 1999; 30(6): 471-477.
9. Abbas GM, Abdel Bar FM, Baraka HN, Gohar AA, Lahloub MF. A new antioxidant
stilbene and other constituents from the stem bark of Morus nigra L. Nat Prod Res, 2014;
28(13): 952-959.
10. Aliaga C, Lissi E. Reactions of the radical cation derived from 2,2`-azinobis(3-
ethylbenzothiazoline-6-sulfonicacid) (ABTS+) with amino acids: kinetics and
mechanism. Canadian Journal of Chemistry, 2000; 78: 1052–1059.
11. Modak B, Rojas M, Torres R, Rodilla J, Luebert F. Antioxidant activity of a new
aromatic geranyl derivative of the resinous exudates from Heliotropium glutinosum Phil.
Molecules, 2007; 12(5): 1057-1063.
12. Kumar RJ, Jyostna D, Krupadanam GLD, Srimannarayana* G. Phenanthrene and
stilbenes from Pterolobium hexapetallum. Phytochemistry, 1988; 27(11): 3625-3626.
13. Li X-C, Elsohly HN, Hufford CD, Clark AM. NMR assignments of ellagic acid
derivatives. Magnetic Resonance in Chemistry, 1999; 37(11): 856-859.
www.wjpr.net Vol 8, Issue 9, 2019.
Fatma. World Journal of Pharmaceutical Research
123
14. Chopra CS, Cole ARH, Theiberg KJL, White DE, Arthur HR. Triterpene compounds—
VII: The constitution of melaleucic acid. Tetrahedron, 1965; 21(6): 1529-1536.
15. Lee CK. New Triterpenes from the Heartwood of Melaleuca leucadendron L. Journal of
the Chinese Chemical Society, 1998; 45: 303-306.
16. Vieira TR, Barbosa LCA, Maltha CRA, Paula VF, Nascimento EA. Chemical
constituents from Melaleuca alternifolia (Myrtaceae). Quimica Nova, 2004; 27(4):
536-539.
17. Tsuruga T, Chun YT, Ebizuka Y, Sankawa U. Biologically active constituents of
Melaleuca leucadendron: inhibitors of induced histamine release from rat mast cells.
Chemical & pharmaceutical bulletin, 1991; 39(12): 3276-3278.
18. Moharram FA, Marzouk MS, El-Toumy SA, Ahmed AA, Aboutabl EA. Polyphenols of
Melaleuca quinquenervia leaves--pharmacological studies of grandinin. Phytotherapy
research: PTR, 2003; 17(7): 767-773.
19. Yaoa L, Jiang Y, Singanusong R, Datta N, Raymont K. Phenolic acids in Australian
Melaleuca, Guioa, Lophostemon, Banksia and Helianthus honeys and their potential for
floral authentication. Food Research International, 2005; 38(6): 651-658.
20. Ali HM, Abo-Shady A, Sharaf Eldeen HA, et al. Structural features, kinetics and SAR
study of radical scavenging and antioxidant activities of phenolic and anilinic compounds.
Chemistry Central journal, 2013; 7(1): 53-53.