Two new phenolic glycosides from the stem of Zanthoxylum armatum DC

Article abstract of DOI:10.1080/14786419.2017.1303695

Two new phenolic glycoside, 2-methoxy-4-hydroxylphenyl-1-O-α-L-rhamnopyranosyl- (1″?→?6′)-β-D-glucopyranoside. (1) and threo-3-methoxy-5-hydroxy-phenylpropanetriol-8-O-β-D-glucopyranoside (2), were isolated from the stems of Zanthoxylum armatum. The compo

Full text of DOI:10.1080/14786419.2017.1303695

Natural Product Research
Formerly Natural Product Letters
ISSN: 1478-6419 (Print) 1478-6427 (Online) Journal homepage: http://www.tandfonline.com/loi/gnpl20
Two new phenolic glycosides from the stem of
Zanthoxylum armatum DC
Tao Guo, Li-Ping Dai, Xiao-Feng Tang, Tong-Tong Song, Ya Wang, Ai-Hong
Zhao, Ying-Ying Cao & Jun Chang
To cite this article: Tao Guo, Li-Ping Dai, Xiao-Feng Tang, Tong-Tong Song, Ya Wang, Ai-
Hong Zhao, Ying-Ying Cao & Jun Chang (2017): Two new phenolic glycosides from the stem of
Zanthoxylum armatum DC, Natural Product Research, DOI: 10.1080/14786419.2017.1303695
To link to this article: http://dx.doi.org/10.1080/14786419.2017.1303695
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Date: 01 April 2017, At: 07:17

Natural Product research, 2017
ꢁꢁp://ꢂx.ꢂꢃi.ꢃꢄg/10.1080/14786419.2017.1303695
ꢀ
Two new phenolic glycosides from the stem of Zanthoxylum
armatum DC
Tao Guoa,b , Li-Ping Dai , Xiao-Feng Tang , Tong-Tong Song , Ya Wang ,
#
c#
b
b
b
b
b
a
Ai-Hong Zhao , Ying-Ying Cao and Jun Chang
ꢅ
b
sꢆꢀꢃꢃꢇ ꢃf Pꢀꢅꢄmꢅꢆy, Fꢈꢂꢅn univꢉꢄꢊiꢁy, sꢀꢅngꢀꢅi, cꢀinꢅ; sꢆꢀꢃꢃꢇ ꢃf lifꢉ sꢆiꢉnꢆꢉ ꢅnꢂ enginꢉꢉꢄing, lꢅnzꢀꢃꢈ
ꢆ
univꢉꢄꢊiꢁy ꢃf tꢉꢆꢀnꢃꢇꢃgy, lꢅnzꢀꢃꢈ, cꢀinꢅ; sꢆꢀꢃꢃꢇ ꢃf Pꢀꢅꢄmꢅꢆy, hꢉnꢅn univꢉꢄꢊiꢁy ꢃf tꢄꢅꢂiꢁiꢃnꢅꢇ cꢀinꢉꢊꢉ
Mꢉꢂiꢆinꢉ, Zꢀꢉngzꢀꢃꢈ, cꢀinꢅ
ABSTRACT
ARTICLE HISTORY
Two new phenolic glycoside, 2-methoxy-4-hydroxylphenyl-1-O-α-
L-rhamnopyranosyl- (1″ → 6′)-β-D-glucopyranoside. (1) and threo-3-
methoxy-5-hydroxy-phenylpropanetriol-8-O-β-D-glucopyranoside
rꢉꢆꢉivꢉꢂ 1 dꢉꢆꢉmbꢉꢄ 2016
aꢆꢆꢉpꢁꢉꢂ 22 Fꢉbꢄꢈꢅꢄy 2017
KEYWORDS
(
2), were isolated from the stems of Zanthoxylum armatum. The
Zanthoxylum armatum;
pꢀꢉnꢃꢇiꢆ gꢇyꢆꢃꢊiꢂꢉ;
ꢅnꢁiꢃxiꢂꢅnꢁ ꢅꢆꢁiviꢁy
compounds 1 and 2 showed weak scavenging activity in DPPH free
radical assay with IC50 values of 323 and 114 mM, respectively.
1
. Introduction
The genus Zanthoxylum is the most widely distributed genus in Rutaceae. It is very well
known for its diversified chemistry, particularly by the presence of alkaloids, aromatic and
aliphatic amides, and phenylpropanoids (Wang et al., 2015). Zanthoxylum armatum DC.,
distributed almost everywhere in china, is a common specie in the genus, and 3–5 m high
of deciduous arbour. Its root, stem, leaf, fruit and seeds were used as herbs, and fruit was
also often used as food condiment and aromatic preservatives. Modern pharmacological
studies have shown that Z. armatum has antispasmodic (Gilani et al. 2010), anti-inflammatory
(Guo et al. 2011), antimicrobial (Mehmood et al. 2013), antitumour (Singh et al. 2015),
CONTACT Jꢈn cꢀꢅng
tꢀꢉ bꢃꢁꢀ ꢅꢈꢁꢀꢃꢄꢊ ꢆꢃnꢁꢄibꢈꢁꢉꢂ ꢉqꢈꢅꢇꢇy ꢁꢃ ꢁꢀiꢊ wꢃꢄk.
jꢆꢀꢅng@fꢈꢂꢅn.ꢉꢂꢈ.ꢆn
#
sꢈppꢇꢉmꢉnꢁꢅꢇ ꢂꢅꢁꢅ fꢃꢄ ꢁꢀiꢊ ꢅꢄꢁiꢆꢇꢉ ꢆꢅn bꢉ ꢅꢆꢆꢉꢊꢊꢉꢂ ꢅꢁ ꢀꢁꢁp://ꢂx.ꢂꢃi.ꢃꢄg/10.1080/14786419.2017.1303695.
©
2017 Infꢃꢄmꢅ uK limiꢁꢉꢂ, ꢁꢄꢅꢂing ꢅꢊ tꢅyꢇꢃꢄ & Fꢄꢅnꢆiꢊ Gꢄꢃꢈp

2
T. GUO ET AL.
Figure 1. cꢀꢉmiꢆꢅꢇ ꢊꢁꢄꢈꢆꢁꢈꢄꢉꢊ ꢃf ꢆꢃmpꢃꢈnꢂ 1 ꢅnꢂ 2.
antidiabetic (Mehmood et al. 2013; Karki et al. 2014), hepatoprotective (Verma & Khosa 2010)
and anthelmintic (Kumar et al. 2016) activities.
Most of the previous phytochemical investigations (Bhatt et al. 2015; Guo et al., 2012,
2015, 2017; Samad et al. 2014) have focused on petroleum ether, ethyl acetate and n-butanol
fractions of Z. armatum extract, while its water fraction of extract was rarely reported. As
part of our systematic investigations on chemical constituents from Z. armatum, the present
study investigated the water fraction of the ethanol extract of Z. armatum, resulting in
isolation of two new compounds, 2-methoxy-4-hydroxylphenyl-1-O-α-L-rhamnopyrano-
syl-(1″→6′)-β-D-glucopyranoside (1) and threo-3-methoxy-5-hydroxy-phenylpropanetriol-
8-O-β-D-glucopyranoside (2) (Figure 1).
2
. Results and discussion
2
0
Compound 1 was obtained as amorphous powder, [ꢀ] −58.2° (H O). The ESI-MS showed
D
2
+
quasimolecular ion peaks at m/z 471 [M + Na] , indicating a molecular weight of 448. Its
1
13
molecular formula was determined to be C H O , according to the HR-ESI-MS, H and C
1
9
28 12
1
NMR spectroscopic data. The H NMR spectrum of 1 showed the presence of a 1,2,4-trisub-
stituted benzene ring [δ 6.98 (1H, d, J = 8.8 Hz), 6.58 (1H, d, J = 2.7 Hz) and 6.42 (1H, dd, J = 8.8,
1
2
.7 Hz)] and a methoxy group [δ 3.80 (3H, s)]. In the sugar part of H NMR spectrum, the
anomeric proton δ 4.88 (1H, d, J = 7.6 Hz) indicated the presence of glucose, while the signal
H
at δ 4.75 (1H, d, J = 1.1 Hz) indicated the presence of rhamnose, which were confirmed by
acid hydrolysis and comparison with authentic samples. The stereochemistry of the anomeric
carbon of glucose was determined as β-configuration according to the coupling constant
of the anomeric proton (J = 7.6 Hz) and the chemical shift of C-1′ (101.6). The anomeric
1
3
carbon of the rhamnosyl residue was determined to be α from the C NMR chemical shifts
of C-3″ and C-5″ (Kasai et al. 1979). The down-field shift of the glucose C-6′ signal (δ 66.6)
C
indicated that glucose and rhamnose were linked through a 1 → 6 glycosidic bond. The
glycosidic site was unambiguously established by a HMBC experiment in which the long-
range correlations between H-1′ (δ 4.88) and C-1 (δ 138.8), H-1″(δ 4.75) and C-6′(δ 66.6) was
observed. Significant HMBC correlations were also observed between OCH and C-2. The
3
site of the methoxy group was further deduced from the results of the NOESY spectrum, in
which NOE signals were only observed between H-1′ (δ 4.88) and H-6 (δ 6.98).
Consequently, the structure of compound 1 was established as 2-methoxy-4-hydroxyl-
phenyl-1-O-α-L-rhamnopyranosyl-(1″ → 6′)-β-D-glucopyranoside.
Compound 2 was obtained as a white amorphous powder. Its molecular formula was
+
1
determined as C H O by HR-ESI-MS at m/z 399.1261 [M + Na] . The H NMR spectrum of
1
6
24 10

NATURAL PRODUCT RESEARCH
3
2
(
(Table S1) showed the presence of three aromatic protons [δ 6.94 (1H, s), 6.81 (1H, s), 6.80
1H, s)] indicating a 1,3,5-trisubstituted benzene ring, two methine protons [δ 4.69 (1H, d,
J = 6.8 Hz), 3.89 (1H, m)], a methylene proton [δ 3.55 (1H, dd, J = 12.3, 3.3 Hz), 3.30 (1H, m)],
1
3
and a methoxy proton [δ 3.76 (3H, s)]. Combining with C NMR spectrum together, the
characteristic resonances of 2 with a phenylpropanoid moiety and a hexose moiety was
suggested. The anomeric proton δ 4.38 (1H, d, J = 7.8 Hz) indicated the presence of glucose,
H
which was confirmed by acid hydrolysis and comparison with authentic samples. The ste-
reochemistry of the anomeric carbon of glucose was determined as β-configuration accord-
ing to the coupling constant of the anomeric proton and the chemical shift of C-1′ (102.8).
The glucose position was established by a HMBC experiment, in which the correlation
between H-1′ and C-8 was observed. The methoxy group was linked to C-3 by the HMBC
correlation between OCH (δ 3.76) and C-3 (δ 147.2). In addition, the coupling constant
3
H
C
1
(
(
J7,8 = 6.8 Hz) in the H NMR spectrum indicated a threo configuration between H-7 and H-8
Chang et al. 1999). Through an analysis of H- H COSY, HMQC and HMBC correlations, the
1
1
compound 2 was determined to be threo-3-methoxy-5-hydroxy-phenylpropanetriol-
8-O-β-D-glucopyranoside.
The results of antioxidant test showed that compounds 1 and 2 displayed some scaveng-
ing activity in DPPH free radical assay with IC50 values of 323 and 114 mM, respectively.
3
. Experimental
.1. General experimental procedures
Optical rotations were measured on a Perkin-Elmer 241 automatic digital polarimeter. UV
3
1
13
spectra were determined with a Shimadzu UV-260 instrument. H, C and 2D NMR spectra
were recorded on a Bruker DRX-400 spectrometer ( H 400 MHz and C 100 MHz) in D O as
1
13
2
the internal standard. Chemical shifts are reported in ppm and coupling constants (J) are
expressed in Hertz. HR-ESI-MS data were acquired on Bruker APEX 7.0TESLA FT-MS apparatus
(
(
Bruker, Axs Gmbh, Switzerland). Reversed-phase chromatography utilised MCI gel CHP 20P
75–150 μm, Mitsubishi Chemical Industries Co., Ltd.), Cosmosil 75 C -OPN (42–105 μm,
1
8
Nacalai Tesque Inc.) columns and TSK gel Toyopearl HW-40F (30–60 μm, Toso Co., Ltd.). TLC
was performed on precoated silica gel 60 F254 plates (0.2 mm, Merck).
3
.2. Plant materials
The stem of Z. armatum DC. was collected from Nanning of Guangxi, China, and was iden-
tified by associate professor Ze-hao Huang. Voucher specimen (#Z140934) was deposited
in school of life science and Engineering, Lanzhou University of Technology, Gansu, China.
3
.3. Extraction and isolation
The air-dried stem of Z. armatum DC.(20 kg) was successively extracted three times (each
for four days) by maceration in 95% ethanol. A residue was obtained after the solvents
evaporation under reduced pressure by a rotary evaporator at 50°C. The residue was sus-
pended in water (2L) and extracted three times with petroleum ether, EtOAc and n-BuOH at
room temperature, successively. The water solvent was evaporated to dryness under reduced

4
T. GUO ET AL.
pressure to afford a brown crude extract (83 g). The extract dissolved in 300 mL water was
subjected to MCI gel CHP 20P (8 × 60 cm) and eluted with MeOH–H O mixtures of decreasing
2
polarity to obtain six fractions as Fr.1 (1.0 L, 100% H O), Fr. 2 (1.0 L, 10% MeOH, v/v), Fr. 3 (1.0
2
L, 30% MeOH, v/v), Fr. 4 (1.0 L, 50% MeOH, v/v), Fr. 5 (1.0 L, 70% MeOH, v/v) and Fr. 6 (1.0 L,
1
00% MeOH). The Fr. 2 (12.9 g) was separated using Toyopearl HW-40F (6 × 60 cm) with H O
2
to give five subfractions (Fr.2A - Fr.2E). The Fr.2A (0.7 g) was further purified by Cosmosil 75
C -OPN (4 × 30 cm, eluted with H O → 10% MeOH) andToyopearl HW-40F (6 × 60 cm, eluted
18
2
with H O) to yield compound 1 (70.5 mg). Fr. 3 (8.9 g) was chromatographed on Toyopearl
2
HW-40F (6 × 60 cm) with H O to obtain five subfractions (Fr.3A - Fr.3D). Fr.3A(0.68 g) was
2
further purified by Cosmosil 75 C -OPN (4 × 30 cm, eluted with H O → 20% MeOH), Toyopearl
18
2
HW-40F (6 × 60 cm, eluted with H O), and Cosmosil 75 C -OPN (4 × 30 cm, eluted with
2
18
10 → 20% MeOH) to yield compound 2 (128.5 mg).
3
.3.1. Compound 1
2
0
+
White amorphous powder;[ꢀ]D −58.2° (c 0.10, H O); HR-ESI-MS m/z 471.1477 [M + Na] (calcd
for C H O Na, 471.1478); H and C NMR: see Table S1.
2
1
13
1
9
28 12
3
.3.2. Compound 2
2
0
+
White amorphous powder;[ꢀ]D −22.5° (c 0.10, H O); HR-ESI-MS m/z 399.1261 [M + Na] (calcd
for C H O Na, 399.1261); H and C NMR: see Table S1.
2
1
13
1
6
24 10
3
.4. Acid hydrolysis
A solution of compounds 1–2 (1 mg each) in 5% HCl (0.5 ml) was heated (90°C) for 2 h. After
removing HCl by evaporation in vacuum, the mixture was diluted with H O and extracted
2
with EtOAc. The aqueous layer was neutralised with 0.1 M NaOH and sugars were detected
by TLC analysis by comparing with authentic sugars: d-glucose and l-rhamnose from 1,
d
-glucose from 2. TLC conditions: CHCl –MeOH–H O (14:6:1), R 0.13 (glucose), R 0.24 (rham-
3 2 f f
nose); n-BuOH-pyridine-H O (6:4:3), R 0.37 (glucose), R 0.47 (rhamnose) (Çalış & Kırmızıbekmez
2
f
f
2004).
3
.5. DPPH radical-scavenging assay
The antioxidant properties were measured using DPPH free radical-scavenging method
reported by our research team (Wang et al. 2014). The concentration required for 50% reduc-
tion (50% scavenging concentration, IC ) of DPPH was determined graphically. Ascorbic
5
0
acid was used as a positive control.
4
. Conclusion
The chemical investigation of the stems of Z. armatum has resulted in the isolation of two
new glycosides, 2-methoxy-4-hydroxy-phenol-1-O-α-L-rhamnopyranosyl-(1″→6′)-β-D-glu-
copyranoside (1) and threo-3-methoxy-5-hydroxy-phenylpropanetriol-8-O-β-D-glucopyra-
noside (2).

NATURAL PRODUCT RESEARCH
5
Supplementary material
Supplementary material relating to this article is available online, alongside Tables S1 and Figure
S1–S13.
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding
This research was supported by the National Natural Science Foundation of China [grant number
81360476].
ORCID
Tao Guo
http://orcid.org/0000-0003-4502-5570
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