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Identification of Phenolic Compounds in Aquilaria crassna Leaves Via Liquid Chromatography-Electrospray Ionization Mass Spectroscopy


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In this study, we extd. Aquilaria crassna with aq. ethanol and water and analyzed the exts. via liq. chromatog. diode array detection and electrospray ionization mass spectrometry (LC-ESI-MS) methods. Phenolics were sepd. using semi-micro HPLC and were identified as iriflophenone 3,5-C-ホイ-diglucoside (1), iriflophenone 3-C-ホイ-glucoside (2), mangiferin (3), iriflophenone 2-O-ホア-rhamnoside (4), genkwanin 5-O-ホイ-primeveroside (5), genkwanin 5-O-ホイ-glucoside (6), genkwanin 4'-Me ether 5-O-ホイ-primeveroside (7), and genkwanin (8) via a comparison with authentic samples. The collision-induced dissocn. (CID)-MS/MS spectra of these polyphenols and the unknown chromatog. peaks were detected using hybrid ion trap time-of-flight (IT-TOF) mass spectrometry. The results of the present study demonstrated that LC-ESI-MS can be useful for the specific quality control of exts. of extd. A. crassna. [on SciFinder(R)]
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Food Sci. Technol. Res., 18 (2), 259262, 2012
Identication of Phenolic Compounds in Aquilaria crassna Leaves Via Liquid
Chromatography-Electrospray Ionization Mass Spectroscopy
Tetsuro iTo
, Mamoru KaKino
, Shigemi TazaWa
, Masayoshi oyama
, Hiroe maruyama
, Yoko araKi
and Munekazu iinuma
Laboratory of Pharmacognosy, Gifu Pharmaceutical University, 1-25-4 Daigaku-nishi, Gifu 501-1196, Japan
Laboratory of Molecular Pharmacology, Gifu Pharmaceutical University, 1-25-4 Daigaku-nishi, Gifu 501-1196, Japan
Nagaragawa Research Center, Api Company Limited, 692-3, Yamasaki, Nagara, Gifu 502-0071, Japan
Received June 19, 2011; Accepted December 19, 2011
In this study, we extracted Aquilaria crassna with aqueous ethanol and water and analyzed the extracts
via liquid chromatography diode array detection and electrospray ionization mass spectrometry (LC–
ESI–MS) methods. Phenolics were separated using semi-micro HPLC and were identied as iriophenone
3,5-C-β-diglucoside (1), iriophenone 3-C-β-glucoside (2), mangiferin (3), iriophenone 2-O-α-rhamnoside
(4), genkwanin 5-O-primeveroside (5), genkwanin 5-O-β-glucoside (6), genkwanin 4′-methyl ether
5-O-β-primeveroside (7), and genkwanin (8) via a comparison with authentic samples. The collision-in-
duced dissociation (CID)-MS/MS spectra of these polyphenols and the unknown chromatographic peaks
were detected using hybrid ion trap time-of-ight (IT-TOF) mass spectrometry. The results of the present
study demonstrated that LC–ESI–MS can be useful for the specic quality control of extracts of extracted
A. crassna.
Keywords: Aquilaria crassna, agarwood leaves, LC–ESI–MS, quality control, polyphenols, laxative effect
*To whom correspondence should be addressed.
Aquilaria is a woody plant in the Thymelaeaceae family
native to Southeast Asia, and some members of the genus,
represented by A. crassna and A. sinensis, are known as
incense trees (Ng, Chang, and Kadir, 1997). The depletion
of wild trees due to indiscriminate cutting of agarwood has
resulted in these trees being listed and protected as an endan-
gered species. In Thailand, A. crassna has been systemati-
cally cultivated to ensure a constant production of resin and
processed for perfume. In addition, the leaves have been
used as tea. The polyphenols present in A. crassna leaves
could be an important source of bioactive components. The
major polyphenols in this material are glycosides of flavo-
noids, benzophenones, and xanthones. Researchers have
recently claried the laxative properties of agarwood leaves
(AL) and identied mangiferin (3) and 5-O-β-primeveroside
(5) as the active components (Hara et al., 2008; Kakino et
al., 2010). The functional food applications of AL require an
exact understanding of not only the chemical components,
but also the AL properties such as their chemical natures,
sizes, solubilities, and the degrees and positions of glycosyl-
ation, which inuence their pharmacokinetics and pharmaco-
dynamics in humans. In addition, it is necessary to identify
the AL phenolics present in these suppliable materials (A.
crassna and A. sinensis) and control the standard qualities of
the extracts prepared via standardized conditions. Due to the
1 : R
= H, R
= R
= β-glucose
: R
= R
= H, R
= β-glucose
: R
= α-rhamnose, R
= R
= H
5 : R
= β-primeverose, R
= H
: R
= β-glucose, R
= H
: R
= β-primeverose, R
= CH
8 : R
= R
= H
phase gradient was used with the percentage of B in A vary-
ing as follows: initial concentration, 10% B; 30 min, 50% B;
and 40 min, 50% B.
IT-TOF MS analysis The diode array analysis was per-
formed on a SPD-M20A (Shimadzu; Kyoto, Japan) scanning
in the range 200 400 nm. CID-MS
experiments were per-
formed on a hybrid IT-TOF mass spectrometer with an ESI
interface (Shimadzu). The negative ESI conditions were as
follows: high voltage probe, −3.5 kV; nebulizing gas flow,
1.5 L/min; CDL temperature, 200; heat block temperature,
200; and drying gas pressure, 200 KPa. CID parameters
were chosen as 70% for the CID energy and 50% for the
collision gas parameter. We used N
gas for CID. The detec-
tor voltage of TOF was 1.6 kV. A solution of triuoroacetic
acid and sodium hydrate was used as the standard sample to
adjust the sensitivity and resolution and to perform the mass
number calibration (ion trap and TOF analyzer).
Results and Discussion
General The common solvents used for the extraction
of phenolic compounds from foods include water, metha-
lack of standard methods for sample preparation, extraction,
and analysis, there is no general consensus on a standard pro-
tocol for the quantitation of phenolic compounds in AL. Our
present study focused only on the identication and not the
quantitation of these phenolic compounds in AL in 60% etha-
nol extracts (ALEE) and hot water extracts (ALWE). The ob-
jective of our study was to identify the phenolic compounds
and generate characteristic chromatographic fingerprints of
ALEE and ALWE via liquid chromatography–electrospray
ionization mass spectrometry (LCESIMS) with multi-
stage analysis (MS
). Our study provides useful information
necessary for the generation of standardized AL materials for
in vitro and in vivo studies and for the authentication of AL-
based food products.
Experimental Methods
General experimental procedures The instruments used
in this study were DGU-20A3, LC-20AP, CBM-20A, SPD-
M20A, and SIL-20A LC instruments for semi-micro HPLC
(Shimadzu, Kyoto, Japan)and a Shimadzu hybrid IT-TOF
mass spectrometer (Shimadzu).
Plant material A. crassna was collected in Pechaboon,
Thailand, in October 2009 and was identied by M. Iinuma,
one of the authors. A voucher specimen has been deposited
at the herbarium of API Co. (Gifu, Japan).
Extraction procedures The dried and chopped leaves of
A. crassna (50 g) were separately extracted with either 60%
(v/v) ethanol (1.0 L, 24 h × 1, room temperature) or water (1.0
L, 1 h × 1, 95), and then were ltered. The extracts were
concentrated in vacuo at 50 to yield alcohol and water AL
extracts [ALEE (11.3 g) and ALWE (11.1 g)], respectively.
Reagents for HPLC All solvents were HPLC grade and
purchased from Sigma Aldrich Co. (St. Louis, MO). EA,
catechin, epicatechin, and quercetin standards were also pur-
chased from Sigma Aldrich Co. Reagent grade formic acid
(98%) was purchased from Nacalai Tesque Inc. (Tokyo, Ja-
pan). Ultra-pure water was prepared using a Millipore Milli-
Q purication system (Bedford, MA).
Sample preparation for LCMS ALEE and ALWE
(each 40.0 mg) were dissolved in 50% ethanol and water (20
mL each), respectively, and injected directly for HPLC−MS
HPLC analysis We performed HPLC analysis using a
Shimadzu HPLC system. Chromatographic separation was
performed on a Capcell Pak UG120 (5 μm, 2.0 i.d. × 250
mm; Shiseido, Tokyo, Japan). Mobile phase A was water
containing 0.1% acetic acid, and mobile phase B was CH
containing 0.1% acetic acid. The column temperature was
40. The HPLC ow rate was 0.2 mL/min. A sample solu-
tion of 1 μL was injected into the HPLC system. A mobile
Fig. 1. HPLC chromatograms of 60% EtOH extract (ALEE) [A]
and hot water extracts (ALWE) [B] on Capcell Pak UG120 (5 μm,
2.0 i.d. × 250 mm) (40). a UV 330 nm, b f Positive ion ESI-
MS chromatograms of total ion current proles (b: 1.0
) and selec-
tive ion modes (c e). [A] c m/z 315 (16.0); d m/z 299 (25.0); e m/z
285 (5.5); f m/z 247 (12.0) [B] c m/z 315 (6.0); d m/z 299 (7.0); e m/
z 285 (2.6); f m/z 247 (9.0).
Relative magnitudes are given in parentheses.
T. iTo et al.260
sults would give promising information for the determination
of the molecular composition, and the negative mode would
provide extensive information via CID fragmentations. The
product ion spectra of the pseudomolecular ions [M+H]+ and
[M−H]− as well as the selective ion chromatograms for the
aglycons, were obtained by conducting CID-MS/MS experi-
ments. The chromatograms of the MS total ion current (TIC)
and the selective ion monitoring (SIM) mode in positive
mode are shown in Fig. 1. The MS
data of 15 are summa-
rized in Table 1.
Identication of 18 by HPLC in ALEE and ALWE
HPLC profiles of ALEE and ALWE monitored at 330 nm
showed that peaks numbered 1 8 were completely sepa-
rated (Fig. 1). These peaks were identied as iriophenone
3,5-C-β-diglucoside (1), iriflophenone 3-C-β-glucoside (2),
nol, aqueous acetone, ethanol, and ethyl acetate (Naczk and
Shahidi, 2004). Because our primary aim is to use AL as a
food material in Japan, we performed the preliminary ex-
tractions with water and various concentrations of aqueous
ethanol at various temperatures. The results of the compre-
hensive HPLC analysis (data not shown) indicated that the
60% aqueous ethanol at room temperature and water at 95
extracts (ALEE and ALWE) were similar and yielded the
most peaks in the HPLC–diode array detection (DAD) chro-
matogram when recorded at 330 nm (Fig. 1, [A] a and [B] a).
The peaks showed absorbance wavelengths (200 400 nm)
typical of phenolics, including benzophenones (1, 2, and 4),
xanthone (3), and avones (5 8) (Fig. 2, a c). However,
80% ethanol is also the accepted solvent for the most effi-
cient extraction to investigate one of the laxative principles
(5) (data not shown). The extraction scheme, facilitated by
high density ethanol, requires custom equipment and is not a
practical method that can be used in API Co. (Gifu, Japan).
We discuss the comparative study of ALEE and ALWE in
this paper.
To obtain good resolution of the peaks in a reasonably
short analysis time, we screened different mobile phases and
semi-micro column compositions. We found that one of the
suitable eluting solvent systems was acetonitrile and 0.1%
aqueous acetic acid, and the suitable column was a Capcell
Pak UG120 (5 μm, 2.0 i.d. × 250 mm; Shiseido; Tokyo,
Japan). The optimized LC conditions permitted good sepa-
ration of eight target polyphenols as well as other phenolic
compounds within 35 min.
The wavelength for the comparison of the chromato-
grams of ALEE and ALWE was 330 nm which is the λmax
of 5, and we performed a DAD analysis, scanning in the
range 190 400 nm. For the MS analysis, we utilized both
positive and negative ion modes of ESI, since dual mode re-
Fig. 2. Three-dimensional HPLC analysis of major chemical compounds in agarwood leaves.
a time, 5.0 40.0 min; wavelength, 190 − 400 nm; intensity max, 180 mAU, b time, 10.0 40.0 min; wavelength, 240 400 nm; intensity
max, 5 mAU, c UV spectra of key peaks [1 (benzophenone), 3 (xanthone), and 5 (avone)].
Table 1. MS
spectra in the positive and negative ion modes of
major polyphenol glycosides in AL.
Rt (min) Isolated m/z MS
type Major product ions (m/z)
1 6.9 571 [M+H]+ MS
535 (100)
, 433 (57)
569 [M−H] MS
449 (21), 431 (13), 359 (57), 329 (100)
449 [MH] MS
329 (100)
359 [MH] MS
239 (100)
2 7.4 409 [M+H]+ MS
391 (100), 325 (64), 313 (64)
407 [MH] MS
287 (100)
3 9.3 423 [M+H]+ MS
369 (17), 357 (17), 351 (26), 327 (41),
303 (21), 299 (17), 273 (100)
421 [MH] MS
331 (49), 301 (100), 271 (23), 259 (37)
310 [MH] MS
273 (25), 272 (25), 258 (100)
4 11.8 393 [M+H]+ MS
339 (100), 247 (62)
339 [M+H]+ MS
247 (100)
391 [MH] MS
245 (100), 151 (15)
245 [MH] MS
151 (100)
5 16.0 579 [M+H]+ MS
285 (100)
577 [MH] MS
283 (100), 268 (24)
283 [MH] MS
268 (100)
Peak numbers are the same as in Fig. 1.
Relative abundances (%) are given in parentheses.
Phenolic Compounds in Agarwood Leaves 261
methoxyavone, MW 284) for 5 and 6, and 5-hydroxy-7,4′-
dimethoxyavone (MW 298) for 7. The SIM chromatograms
in Fig. 1 present those of [M+H]+ at m/z 299 (d), 285, (e)
and 247 (f) in addition to that of the noticeable ion at m/z
315 (c). It was evident that 4 7 produced product ion peaks
of their aglycons. In addition, we also detected some O-
glucosides hitherto unidentied in the SIM chromatograms,
exemplied by the aglycons with [M+H]+ at m/z 315 (c) and
247 (f). The composition of the SIM peak for [M+H]+ at m/
z 315 at 33.6 min (Fig. 1, [A] c) was calculated for C
with a pseudomolecular ion [M+H]+ at m/z 315.0878 (calcd.
315.0863), corresponding to dihydroxydimethoxyflavone
and adding a further ngerprint to AL.
In conclusion, we have identified the major phenolic
compounds present in agarwood leaves (AL) and established
their characteristic chromatographic profiles and composi-
tional similarities for 60% ALEE and ALWE. Among the
identified compounds, genkwanin 5-O-β-primeveroside (5)
and mangiferin (3) showed laxative activities via different
Because it is meaningful to evaluate chemical composi-
tional similarities and biological properties of various extract
forms, these methods should aid in the standardization of AL
materials for in vitro and in vivo studies. The chromatograph-
ic ngerprinting of AL should also be useful for the authenti-
cation of AL-based food products. The information provided
by our study will aid in the evaluation of the importance of
AL consumption on human health.
Ng, L.T., Chang, Y.S. and Kadir, A.A. (1997) A review on agar
(gaharu) producing Aquilaria species. J. Tropic. Forest Prod., 2,
Hara, H., Ise, Y., Morimoto, N., Shimazawa, M., Ichihashi, K.,
Ohyama, M. and Iinuma, M. (2008) Laxative effect of agarwood
leaves and its mechanism. Biosci. Biotechnol. Biochem., 72, 335-
Kakino, M., Izuta, H., Ito, T., Tsuruma, K., Araki, Y., Shimazawa,
M., Oyama, M., Iinuma, M. and Hara, H. (2010) Agarwood in-
duced laxative effects via acetylcholine receptors on loperamide-
induced constipation in mice. Biosci. Biotechnol. Biochem., 74,
Naczk, M. and Shahidi, F. (2004) Extraction and analysis of pheno-
lics in food. J. Chromat. B, 1054, 95-111.
mangiferin (3), iriflophenone 2-O-α-rhamnoside (4), genk-
wanin 5-O-β-primeveroside (5), genkwanin 5-O-β-glucoside
(6), genkwanin 4′-methyl ether 5-O-β-primeveroside (7), and
genkwanin (8) by comparison with standard samples. The
composition of ALEE (Fig. 1, [A]) and ALWE (Fig. 1, [B])
were similar except for the ratios of representative polyphe-
nols (1 8). Because our study focused on identication and
not quantitation, the differences in the ratio of each polyphe-
nol with the peak of 3 were discussed. For quality control of
the AL extracts, it is important to identify exact information
on the bioactive polyphenols bearing laxative properties (3
and 5). We found evidence for the existence of both com-
pounds in both extracts as well as the effective extraction of
5 with hot water, strongly indicating that both extracts are
substantial functional food materials with signicant laxative
effects. The other evident differences were the ratios of the
other polyphenols such as 1 (ALEE < ALWE), 2 (ALEE <
ALWE), 7 (ALEE < ALWE), and 8 (ALEE > ALWE). These
differences were acceptable when we considered the struc-
tures of 18 and the solvents used for the extraction. Pheno-
lic glycosides, represented as 1 7 in AL, are well known to
dissolve in hot water. In contrast, genin (8) was more soluble
in organic solvents, thus reecting the analytical results.
DAD analysis of major polyphenols in the ALEE
Three-dimensional HPLC analysis showed absorbance wave-
lengths typical of phenolic compounds. Figure 2 presents the
UV spectra of three majorities (Fig. 2 a, maximum mAU:
180) and the others (Fig. 2 b, maximum mAU: 5), as well as
UV spectra of 1, 3, and 5 (Fig. 2c). The results indicated that
the LC−DAD ngerprint of the AL extract existed as three
major peaks of two benzophenones (1 and 2) and xanthone (3)
and the other minor peaks of benzophenones (4) and avones
(5 and 8).
ESI-MS/IT-TOF analysis of ALEE and ALWE Figure
1 shows the chromatograms of TIC and SIM in the positive
mode (A, ALEE and B, ALWE). Not all compounds in AL
were detected in the DA detector, e.g., due to lack of conju-
gated systems in some compounds in the extract. However,
the TIC chromatograms of ALEE (Fig. 1, [A] b) and ALWE
(Fig. 1, [B] b) showed superimposable features with those
of the respective chromatograms at 330 nm (a) except for
the initial 5 min. The TIC chromatograms also supported the
compositional similarity of ALEE and ALWE.
The major phenolic compounds in AL existed as C-glu-
cosides (1 3) and O-glucosides (4 7), and the aglycons
of 4 7 were iriflophenone (1,3,4,4-tetrahydroxy benzo-
phenone, MW 246) for 4, genkwanin (8: 5,4′-dihydroxy-7-
T. iTo et al.262
... The compound is annotated as aquisiflavoside, which was isolated previously from A. sinensis leaves and was reported as potent nitric oxide inhibitor [62]. The fragmentation pattern of tentatively identified aquisiflavoside (Supplementary Figure S30 Figure S31), as previously reported in different Aquilaria species [63,64], while peak C37 (m/z 299.0561, C16H11O6 − ) expressed the presence of extra hydroxyl group to compound 40, which suggests that C37 is Hydroxygenkwanin (Supplementary Figure S32). ...
... Figure S37). Mangiferin was previously identified in Aquilaria leaves [36,63,64]. Peak C9 (m/z 435.0979, ...
Full-text available
The Aquilaria malaccensis species of the genus Aquilaria is an abundant source of agarwood resin and many bioactive phytochemicals. Recent data regarding the chemical constituents and biological activities of Aquilaria leaves led us to attempt to qualitatively profile the metabolites of Aquilaria malaccensis leaves from a healthy, noninoculated tree through phytochemical screening, GC-MS, and LC/Q-TOF-MS. The present work is also the first to report the antilipoxygenase activity of A. malaccensis leaves from healthy noninoculated tree and investigate its toxicity on oral mucosal cells. A total of 53 compounds were tentatively identified in the extract, some of which have been described in literature as exhibiting anti-inflammatory activity. A number of novel compounds were identified for the first time, including quercetin, quercetin-O-hexoside, kaempferol-O-dirhamnoside, isorhamnetin-O-hexoside, syringetin-O-hexoside, myricetin, tetrahydroxyflavanone, hesperetin, sissotrin, and lupeol. The antilipoxygenase assay was used to determine the lipoxygenase (LOX) inhibitory potential of the extract, while a WST-1 assay was conducted to investigate the effect of the extract on oral epithelial cells (OEC). The extract implied moderate anti-LOX activity with IC50 value of 71.6 μg/mL. Meanwhile, the cell viability of OEC ranged between 92.55% (10 μg/mL)-76.06% ± (100 μg/mL) upon treatment, indicating some potential toxicity risks. The results attained encourage future studies of the isolation of bioactive compounds from Aquilaria malaccensis leaves, as well as further investigation on the anti-inflammatory mechanisms and toxicity associated with their use.
... Compound 3 was characterized by m/z 409.1107 and 407.1012 in positive and negative mode, respectively, suggesting a molecular formula of C 19 H 20 O 10 . The UV absorption spectrum (λ max at 296 nm) and complex MS 2 fragmentation suggest that 3 is a glycosylated benzophenone-C-heteroside, tentatively identified as iriflophenone 3-C-β-glucoside [38]. By comparison with standards, peaks 4 and 8 were identified as (+)-catechin and (−)-epicatechin, respectively, and peak 10 as mangiferin (Mang Its UV spectrum (λ max at 265 and 330 nm) resembles a di-hexosyl flavone [39]. ...
This work has two major goals: clarifying in what abiotic stress responses the phenolic compounds, and more particularly, the chlorogenic acid 5-CQA, were involved in C. arabica leaves and developing an innovative tool to study the cellular mechanisms by which 5-CQA intervene during stress response.To achieve the first goal, coffee plants have been grown in field or in controlled conditions and the phenolic content of mature leaves in different stress conditions have been analyzed. A previous study comparing leaves grown in field and in controlled conditions showed that leaves from field contained a greater number of phenolic compounds (17 vs 12 in controlled conditions) and that glycosylated flavonoids are quasi absent in leaves grown under controlled conditions. In field, stresses were applied by growing at full sun or under shade 35 genotypes, including Ethiopian wild accessions, American pure lines and their hybrids, in four different locations in America. Results show that 5-CQA, F-dihex, mangiferin and rutin are four phenolics that could still predict the degree of adaptation to full sunlight or shade. Two of them, 5-CQA and mangiferin appeared as genetic biomarkers, able to differentiate Ethiopian wild accessions from American pure lines. In controlled conditions, all the major phenolic compounds, including 5-CQA and 3,5-diCQA are involved in the response to variation in light intensity, N deprivation, moderate elevation of the thermal regime (mimicking climate changes) or heat wave stress. This suggests that 5-CQA has a central role in the response of Arabica to stress I controlled conditions. However, the absence of glycosylated flavonoids, involved in stress response in field, may modify the phenolic profile of the coffee plant response to abiotic stresses..The second goal proposes to use nanoparticles to make entering 5-CQA into the nucleus of coffee cells. The first step was to obtain nanoparticles. Prussian-blue nanoparticles (PB-NPs) with a cubic shape of 70 nm were synthetized and 2-aminoanthracene was grafted on their structure to make them observable at 504 nm with a confocal microscope. Using calli culture of coffee plants, it was shown that PB-NPs were unable to penetrate into the cells by passive transport. By preparing coffee protoplasts, penetration of nanoparticles was observable, beginning by the invagination of the protoplast membrane. Among different treatments used to facilitate the penetration, electroporation provides the best results, without altering the viability of the protoplasts. These initial results lead to nanoparticle usage for transporting targeted molecules into plant cells. The enrichment of nanoparticles in 5-CQA for its release in the protoplasts will constitute the continuation of this work and will make it possible to better understand the mode of action of 5-CQA in the stress response.
... 14 Mangiferin, genkwanin, and iriflophenone glycosides were generally found in A. crassna and A. sinensis leaves when extracted in ethanol using the maceration method and detected using liquid chromatography-mass spectrometry (LC-MS) and silica gel column chromatography (SGCC). 13,[15][16][17][18][19] Utilization of these leaves requires research series for future development, especially related to raw materials derived from cultivation. Fundamental research that needs to be done is to search for any chemical components contained in the agarwood leaves. ...
... Compound 3 was characterized by m/z 409.1107 and 407.1012 in positive and negative mode, respectively, suggesting a molecular formula of C19H20O10. The UV absorption spectrum (λmax at 296 nm) and complex MS 2 fragmentation suggest that 3 is a glycosylated benzophenone-C-heteroside, tentatively identified as iriflophenone 3-C-β-glucoside [38]. By comparison with standards, peaks 4 and 8 were identified as (+)-catechin and (−)-epicatechin, respectively, and peak 10 as mangiferin (Mang). ...
Full-text available
Phenolic compounds are involved in plant response to environmental conditions and are highly present in leaves of Coffea arabica L., originally an understory shrub. To increase knowledge of C. arabica leaf phenolic compounds and their patterns in adaptation to light intensity, mature leaves of Ethiopian wild accessions, American pure lines and their relative F1 hybrids were sampled in full sun or under 50% shade field plots in Mexico and at two contrasting elevations in Nicaragua and Colombia. Twenty-one phenolic compounds were identified by LC-DAD-MS 2 and sixteen were quantified by HPLC-DAD. Four of them appeared to be involved in C. arabica response to light intensity. They were consistently more accumulated in full sun, presenting a stable ratio of leaf content in the sun vs. shade for all the studied genotypes: 1.6 for 5-CQA, F-dihex and mangiferin and 2.8 for rutin. Moreover, 5-CQA and mangiferin contents, in full sun and shade, allowed for differentiating the two genetic groups of Ethiopian wild accessions (higher contents) vs. cultivated American pure lines. They appear, therefore, to be potential biomarkers of adaptation of C. arabica to light intensity for breeding programs. We hypothesize that low 5-CQA and mangiferin leaf contents should be searched for adaptation to full-sun cropping systems and high contents used for agroforestry systems.
... In addition, much attention has been paid to phytochemical investigations and properties of active ingredients from these plants in recent years. Aquilaria crassna has been identified as a convenient and useful source of biological functional components, such as phenolic, flavonoids, benzophenones, xanthones and sesquiterpenes [13]. In fact, several studies have focused on the biological activities of Aquilaria crassna. ...
... In addition, much attention has been paid to phytochemical investigations and properties of active ingredients from these plants in recent years. Aquilaria crassna has been identified as a convenient and useful source of biological functional components, such as phenolic, flavonoids, benzophenones, xanthones and sesquiterpenes [13]. In fact, several studies have focused on the biological activities of Aquilaria crassna. ...
Full-text available
Aquilaria Crassna an important medicinal plant. It is one of the most growing species of the Thymelaeaceae family. It's found in rainforests of Southeast Asia such as Indonesia, Bangladesh, Malaysia, Thailand, Cambodia, Laos, Vietnam, and Northeastern India. It is a valuable plant on earth because of its wide medicinal properties. Agarwood is consumed as a pathological product as well as it is also an important ingredient in medicine. This review provides significant information regarding the medicinal properties of drugs for the treatment of various diseases such as insomnia, stress, skin damage, stork, arthritis, vomiting, cardiac disorder, cough, asthma, heart attack, kidney failure and agarwood as a treatment for cirrhosis of the liver and as the director or focus of other drugs. It has been used to treat lung and stomach tumors. The plant has several pharmacological activities known as anti-oxidant, antinociceptive, laxative, sedative, antihyperglycemic, thrombolytic, antibiotic, anti-ulcer protective. Each part of the agarwood (Aquilaria Crassna) tree has beneficial properties that can serve mankind. The whole plant can be extensively studied for future possibilities. This review highlights to Pharmacological activity of agarwood (Aquilaria Crassna) in human life and medical benefits at modern civilization.
... Previous phytochemical studies of this genus showed the presence of terpenoids, phenolic compounds, flavonoids and sterols that contribute to its biological activities [9]. The leaves of Aquilaria sp. are believed to have major polyphenolic compounds which can be used as herbal tea and showed a laxative [10][11][12], anti-bacterial [13], and anti-diabetic properties [7]. In addition, stem of A. subintegra has been reported as promising in Alzheimer therapy [14]. ...
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Obesity is one of the serious health condition in the world and contributes to many chronic illnesses augmenting to high risk of death rate. With the purpose of obesity treatment, natural sources come to be significant to replace commercial drugs due to their harmful effect to the body. Therefore, a lot of studies were conducted on natural sources due to their potential as medicinal herbs. Aquilaria sp. plants are well known in traditional medicine as a sedative, analgesic and digestive. Even though there were no reports on applications of Aquilaria sp. towards obesity treatment, this plant has traditionally been used as laxative for weight reducing. In this study, the anti-lipase activity of an extract of barks and leaves from A. subintegra and A. malaccensis were investigated. The anti-lipase activity was measured by colorimetric assay on pancreatic lipase activity using porcine pancreatic lipase (PPL; triacylglycerol lipase, EC The result indicated that among these two species of Aquilaria, A. malaccensis bark in dichloromethane crude showed high anti-lipase activity. Thus, these results suggest that Aquilaria sp. plant extracts might be of therapeutic interest with respect to the treatment.
Covering: Up to the end of 2019. Agarwood is a resinous portion of Aquilaria trees, which is formed in response to environmental stress factors such as physical injury or microbial attack. It is very sought-after among the natural incenses, as well as for its medicinal properties in traditional Chinese and Ayurvedic medicine. Interestingly, the chemical constituents of agarwood and healthy Aquilaria trees are quite different. Sesquiterpenes and 2-(2-phenethyl)chromones with diverse scaffolds commonly accumulate in agarwood. Similar structures have rarely been reported from the original trees that mainly contain flavonoids, benzophenones, xanthones, lignans, simple phenolic compounds, megastigmanes, diterpenoids, triterpenoids, steroids, alkaloids, etc. This review summarizes the chemical constituents and biological activities both in agarwood and Aquilaria trees, and their biosynthesis is discussed in order to give a comprehensive overview of the research progress on agarwood.
Conference Paper
Aquilaria crassna is an Asian traditional plant with diverse Pharmaceutical and industrial properties. Different extraction methods affect the yield and functional activities of the plant products. The current study was designed to measure the performances of five extraction methods (supercritical-fluid extraction SFE, hydrodistillation, steamdistillation, n-hexane and ethanol) on agarwood samples. The stem bark extracts were subjected for anti-proliferative effect using common cancer cell lines (colorectal, pancreatic, prostate and breast). In addition, the scavenging activity was evaluated using two methods (diphenylpicrylhydrazyl) (DPPH) and ferric reducing antioxidant power (FRAP). The present of essential oils was detected by transmission electron microscopy (TEM). The hydrodistillation and (SFE) extracts provided the highest yield with significant antioxidant and anticancer results.
Introduction: Recently, the fresh leaves of Aquilaria trees have been processed as food products such as agarwood tea due to its beneficial medicinal properties. However, there have not been any reported analytical methods to quantify the bioactive principles in the processed products. Objective: A rapid and sensitive ultrahigh-performance liquid chromatography (UHPLC) coupled with electrospray ionisation (ESI) tandem mass spectrometry (MS/MS) method was developed and validated for the simultaneous determination of 10 bioactive components in Aquilaria leaf tea. Methods: The UHPLC-MS/MS was used for quantification operated in multiple reaction monitoring (MRM) mode. The optimised chromatographic parameters were conducted on a Shim-pack XR-ODS II column and mobile phases consisted of acetonitrile and 0.1% formic acid in water. Results: All the samples were analysed within 20 min. The established method showed excellent linearity (R2 > 0.9988), good repeatability with all the relative standard deviation values lower than 3.27%, and satisfactory recovery (79.72-119.22%). The matrix effect factors ranged from 87.65 to 97.27% in the examination. The developed method was applied to the determination of 10 bioactive principles (1-10) in six different Aquilaria leaf tea samples. Among the analytes, mangiferin (1) and iriflophenone 2-O-α-l-rhamnopyranoside (2) were the most abundant compounds in the extracts of Aquilaria leaf tea, and it indicated that these biotech products may possess laxative effects. Conclusion: This proposed method appeared to be a useful tool for the quality control of commercial products of Aquilaria leaf tea and thus provided an analytical reference for herbal drinks.
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Agarwood (Aquilaria sinensis, Aquilaria crasna) is well known as an incense in the oriental region such as Thailand, Taiwan, and Cambodia, and is used as a digestive in traditional medicine. We investigated the laxative effects and mechanism of agarwood leaves extracted with ethanol (EEA-1, Aquilaria sinensis; EEA-2, Aquilaria crasna). EEA-1, EEA-2, the main constituents of EEAs (mangiferin, and genkwanin-5-O-primeveroside), and senna increased the frequency and weight of stools in loperamide-induced constipation model mice. EEA-1 and EEA-2 did not induce diarrhea as a side effect, but senna induced severe diarrhea. EEA-1 and senna increased gastro-intestinal (GI) transit in the model mice. EEA-1, but not senna, also increased the intestinal tension of isolated jejunum and ileum in guinea pigs, and the tension increase was blocked by atropine, a muscarinic receptor antagonist, but not by other inhibitors (granicetron, pyrilamine, or bradykinin-antagonist peptide). Furthermore, the increase in frequency and weight of stools induced by EEA-1 were blocked by pre-administration of atropine in the model mice. These findings indicate that EEAs exerted a laxative effect via acetylcholine receptors in the mouse constipation model.
Phenolics are ubiquitous compounds found in all plants as their secondary metabolites. These include simple phenols, hydroxybenzoic acid and cinnamic acid derivatives, flavonoids, coumarines and tannins, among others. The extraction of phenolics from source materials is the first step involved in their analysis. While chemical methods are used for determination of total content of phenolics, chromatographic and spectrometric analyses are employed for identification and quantification of individual compounds present. This paper provides a summary of background information and methodologies used for the analysis of phenolics in foods and nutraceuticals.
We investigated the laxative activity of an extract of agarwood leaves from Aquilaria sinensis. The laxative activity was measured in mice by counting the stool frequency and stool weight, and the drugs were orally administered. An acetone extract of agarwood leaves and senna (a representative laxative drug) both increased the stool frequency and weight, but a methanol extract did not. The laxative effect of the acetone extract was milder than that of the anthraquinoid laxative, senna, and the former did not induce diarrhea as a severe side effect. We identified the main constituent contributing to the laxative effect of the acetone extract as genkwanin 5-O-beta-primeveroside (compound 4). Compound 4 strengthened the spontaneous motility and induced contraction in the ileum. This ileal contraction induced by compound 4 was inhibited by atropine, but not by azasetron, suggesting that the effect of compound 4 was mediated by acetylcholine receptors, and not by serotonin. The laxative mechanism for compound 4 may in part involve stimulation of intestinal motility via acetylcholine receptors.
A review on agar (gaharu) producing Aquilaria species Laxative effect of agarwood leaves and its mechanism Agarwood in-duced laxative effects via acetylcholine receptors on loperamide-induced constipation in mice
  • L T Ng
  • Y S Chang
  • A A Kadir
  • H Hara
  • Y Ise
  • N Morimoto
  • M Shimazawa
  • K Ichihashi
  • M Ohyama
  • M Iinuma
Ng, L.T., Chang, Y.S. and Kadir, A.A. (1997) A review on agar (gaharu) producing Aquilaria species. J. Tropic. Forest Prod., 2, 272-285. Hara, H., Ise, Y., Morimoto, N., Shimazawa, M., Ichihashi, K., Ohyama, M. and Iinuma, M. (2008) Laxative effect of agarwood leaves and its mechanism. Biosci. Biotechnol. Biochem., 72, 335- 345. Kakino, M., Izuta, H., Ito, T., Tsuruma, K., Araki, Y., Shimazawa, M., Oyama, M., Iinuma, M. and Hara, H. (2010) Agarwood in-duced laxative effects via acetylcholine receptors on loperamide-induced constipation in mice. Biosci. Biotechnol. Biochem., 74, 1550-1555