Two new lignan glycosides from the fruits of Pyrus ussuriensis

Article abstract of DOI:10.1080/10286020.2016.1210132

Two new lignan glycosides, ussuriensislignan A (1) and ussuriensislignan B (2), together with seventeen known compounds (3–19), were isolated from the fruits of Pyrus ussuriensis. Their structures were determined by various spectroscopic methods. This is the first report of the isolation of lignans (compounds 1–3) from the genus Pyrus, and compounds 3–6, 12–16 were reported from Pyrus for the first time.

Full text of DOI:10.1080/10286020.2016.1210132

Journal of Asian Natural Products Research
ISSN: 1028-6020 (Print) 1477-2213 (Online) Journal homepage: http://www.tandfonline.com/loi/ganp20
Two new lignan glycosides from the fruits of Pyrus
ussuriensis
Ze-Qing Zhao, Yan-Fang Su, Fei Yang, Xiu-Mei Gao & Tian-Xiang Li
To cite this article: Ze-Qing Zhao, Yan-Fang Su, Fei Yang, Xiu-Mei Gao & Tian-Xiang Li (2016):
Two new lignan glycosides from the fruits of Pyrus ussuriensis, Journal of Asian Natural
Products Research, DOI: 10.1080/10286020.2016.1210132
To link to this article: http://dx.doi.org/10.1080/10286020.2016.1210132
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Date: 02 August 2016, At: 00:04

Journal of asian natural Products research, 2016
ꢁꢁp://ꢂx.ꢂꢃꢄ.ꢃꢅg/10.1080/10286020.2016.1210132
ꢀ
Two new lignan glycosides from the fruits of Pyrus ussuriensis
a
a,b
a
c
c
Ze-Qing Zhao , Yan-Fang Su , Fei Yang , Xiu-Mei Gao and Tian-Xiang Li
ꢆ
tꢄꢆꢇjꢄꢇ Kꢈy lꢆbꢃꢅꢆꢁꢃꢅy ꢉꢃꢅ Mꢃꢂꢈꢅꢇ dꢅꢊg dꢈꢋꢄvꢈꢅy ꢆꢇꢂ hꢄgꢀ-effiꢌꢄꢈꢇꢌy, sꢌꢀꢃꢃꢋ ꢃꢉ Pꢀꢆꢅmꢆꢌꢈꢊꢁꢄꢌꢆꢋ sꢌꢄꢈꢇꢌꢈ ꢆꢇꢂ
b
tꢈꢌꢀꢇꢃꢋꢃgy, tꢄꢆꢇjꢄꢇ uꢇꢄvꢈꢅꢍꢄꢁy, tꢄꢆꢇjꢄꢇ 300072, cꢀꢄꢇꢆ; cꢃꢋꢋꢆbꢃꢅꢆꢁꢄvꢈ iꢇꢇꢃvꢆꢁꢄꢃꢇ cꢈꢇꢁꢈꢅ ꢃꢉ cꢀꢈmꢄꢌꢆꢋ sꢌꢄꢈꢇꢌꢈ
ꢆꢇꢂ eꢇgꢄꢇꢈꢈꢅꢄꢇg (tꢄꢆꢇjꢄꢇ), tꢄꢆꢇjꢄꢇ 300072, cꢀꢄꢇꢆ; sꢌꢀꢃꢃꢋ ꢃꢉ cꢀꢄꢇꢈꢍꢈ Mꢆꢁꢈꢅꢄꢆ Mꢈꢂꢄꢌꢆ, tꢄꢆꢇjꢄꢇ uꢇꢄvꢈꢅꢍꢄꢁy ꢃꢉ
ꢌ
tꢅꢆꢂꢄꢁꢄꢃꢇꢆꢋ cꢀꢄꢇꢈꢍꢈ Mꢈꢂꢄꢌꢄꢇꢈ, tꢄꢆꢇjꢄꢇ 300193, cꢀꢄꢇꢆ
ABSTRACT
ARTICLE HISTORY
rꢈꢌꢈꢄvꢈꢂ 26 oꢌꢁꢃbꢈꢅ 2015
aꢌꢌꢈpꢁꢈꢂ 3 Jꢊꢋy 2016
Twonewlignanglycosides,ussuriensislignanA(1)andussuriensislignan
B (2), together with seventeen known compounds (3–19), were
isolated from the fruits of Pyrus ussuriensis. Their structures were
determined by various spectroscopic methods. This is the first report
of the isolation of lignans (compounds 1–3) from the genus Pyrus, and
compounds 3–6, 12–16 were reported from Pyrus for the first time.
KEYWORDS
rꢃꢍꢆꢌꢈꢆꢈ; Pyrus ussuriensis;
ꢋꢄgꢇꢆꢇ gꢋyꢌꢃꢍꢄꢂꢈꢍ
1
. Introduction

e fruits of Pyrus ussuriensis (Rosaceae), widely distributed in North China, are popular for
their exceptional fragrance, sweetness, juiciness, and richness of nutrients. e fresh fruits
(Qiuzi pear) can be used for prevention and treatment of some ailments, such as coughs
and colds. Previous phytochemical studies on the genus Pyrus have led to the isolation of
flavonoids, phenolic acids, and triterpenoids [1–3]. In order to better understand the func-
tional properties of the fruits and provide evidence for the development of functional food
products with this plant, a systematic study was carried out to investigate its antioxidant
components of the fruits. Herein, the isolation and structure elucidation of two new lignan
glycosides (1–2), together with seventeen known compounds (3–19), are reported from
the fruits of P. ussuriensis (Figure 1).
2
. Results and discussion
20
Compound 1 was isolated as an amorphous powder with [ꢀ] −5.26 (c 0.57, MeOH). Its
D
molecular formula was determined to be C H O based on the pseudo-molecular ion at
2
8
36 14
+
1
m/z 619.1990 [M+Na] in the HRESIMS experiment. e H NMR spectrum displayed two
singlet signals at δ 6.78 (2H, H-2, H-6) and δ 6.71 (2H, H-2′, H-6′) attributed to aromatic
1
3
protons. e C NMR spectrum showed two benzene rings, six characteristic aliphatic
carbon signals at δ 87.4 and 89.4 (C-7 and C-7′), δ 62.7 and 92.9 (C-8 and C-8′), and δ 72.2
and 76.3 (C-9 and C-9′), four aromatic methoxy groups and resonances corresponding to the
1
13
presence of a single sugar moiety. Assignments of all H and C NMR spectroscopic signals
CONTACT Yꢆꢇ-fꢆꢇg sꢊ
ꢍꢊyꢆꢇꢉꢆꢇg@ꢁjꢊ.ꢈꢂꢊ.ꢌꢇ
tꢀꢈ ꢍꢊppꢋꢈmꢈꢇꢁꢆꢅy ꢂꢆꢁꢆ ꢉꢃꢅ ꢁꢀꢄꢍ ꢆꢅꢁꢄꢌꢋꢈ ꢄꢍ ꢆvꢆꢄꢋꢆbꢋꢈ ꢃꢇꢋꢄꢇꢈ ꢆꢁ ꢀꢁꢁp://ꢂx.ꢂꢃꢄ.ꢃꢅg/10.1080/10286020.2016.1210132.
©
2016 iꢇꢉꢃꢅmꢆ uK lꢄmꢄꢁꢈꢂ, ꢁꢅꢆꢂꢄꢇg ꢆꢍ tꢆyꢋꢃꢅ & fꢅꢆꢇꢌꢄꢍ Gꢅꢃꢊp

2
Z.ꢀQ. ZꢁAꢂ ꢃT AL.
were based on COSY, HSQC, HMBC, and NOESY experiments. e presence of a β-ꢀ-glu-
copyranosyl moiety in the molecule was confirmed by an anomeric proton signal at δ 4.86
(
1H, d, J = 7.8 Hz) and six carbon signals at δ 105.3, 75.7, 77.8, 71.3, 78.4, and 62.6 [4], along
with acid hydrolysis of 1. e ꢀ configuration of the glucose was assumed from biogenetic
consideration [5]. e 1D NMR spectra (Table 1) revealed the presence of 7,9′:7′,9-diepox-
ylignan skeleton (Figure 1). e position of the glucose moiety at C-4 was confirmed by
an HMBC correlation between H-1″ (δ 4.86) and C-4 (δ 135.7). e cross peaks observed
in the HMBC spectrum between H-7 (δ 4.88) and C-8/C-8′/C-9/C-9′, H-7′ (δ 4.65) and
C-8′/C-9′, H-8 (δ 2.98–3.04) and C-8′, and H -9, H -9′ and C-7/C-7′/C-8/C-8′ unambigu-
2
2
ously provided evidence of the 3,7-dioxabicyclo[3.3.0]octane structure. e HMBC corre-
lations between H-7 and C-1/C-2/C-6 established that ring A was at the C-7 position, and
the cross peaks between H-7′ and C-1′/C-2′/C-6′ determined that ring B was attached to
C-7′ (Figure 2). e two aryl groups of 1 were all in equatorial positions supported by the
chemical shif s of C-7 and C-7′ (δ 87.4 and 89.4, respectively) [5]. e H-7/H-8 relative
configuration was assigned as trans from the small coupling constant between H-7 and H-8
(
J7, 8 = 4.8 Hz) [6]. e NOE correlations between H-8 and H-2/H-6 indicated that H-8 and
ring A were positioned on the same side (Figure 2). Natural tetrahydrofuranoid lignans
observed in the literature were usually cis-8,8′-fused furfuran lignans [6], and the carbon
chemical shif s of compound 1 at the stereogenic centers were similar to those of fraxire-
sinol-4′-O-β-ꢀ-glucopyranoside [4], which demonstrated that they have the same relative
configuration. Meanwhile, the CD data and specific rotation of 1 were consistent with those
of fraxiresinol-4′-O-β-ꢀ-glucopyranoside [4] and hydroxypinoresinol-4′-O-β-ꢀ-glucopyra-
noside [7], suggesting that they possessed the same absolute configuration. In view of all the
above evidence, compound 1 was determined as (–)-7S,7′R,8R,8′S-3,3′,5,5′-tetramethoxy-
4
′,8′-dihydroxy-7,9′:7′,9- diepoxylignan-4-O-β-ꢀ-glucopyranoside and given the trival
name ussuriensislignan A.
20
Ussuriensislignan B (2) was obtained as an amorphous powder with[ꢀ] −3.70 (c 0.27,
D
−
MeOH). A molecular formula of C H O was determined from the [M−H] ion peak at
2
8
36 14
1
13
m/z 595.2027, which was the same as that of 1. e H and C NMR spectroscopic data
of compound 2 (Table 1) indicated the presence of one glucose, two benzene rings, one
3
,7-dioxabicyclo[3.3.0]octane unit, and four aromatic methoxy groups in the molecule
(
Figure 1). Comparison of NMR spectroscopic data of 1 and 2 indicated that they possessed
similar structures. e upfield shif s for C-1 (Δδ 6.4 ppm), C-3/C-5 (Δδ 5.1 ppm), and
Figure 1. sꢁꢅꢊꢌꢁꢊꢅꢈꢍ ꢃꢉ ꢌꢃmpꢃꢊꢇꢂꢍ 1–2.

JꢂꢄꢅꢆAL ꢂF ASꢇAꢆ ꢆATꢄꢅAL PꢅꢂꢈꢄꢉTS ꢅꢃSꢃAꢅꢉꢁ
3
1
13
Table 1. h (600 Mhz) ꢆꢇꢂ c (150 Mhz) nMr ꢍpꢈꢌꢁꢅꢃꢍꢌꢃpꢄꢌ ꢂꢆꢁꢆ ꢉꢃꢅ ꢌꢃmpꢃꢊꢇꢂꢍ 1−2 ꢄꢇ cd od.
3
1
2
Position
δꢁ (J in ꢁz)
δꢉ
δꢁ (J in ꢁz)
δꢉ
1
2
3
4
5
6
7
8
139.3
105.3
154.4
135.7
154.4
105.3
87.4
132.9
105.0
149.3
136.3
149.3
105.0
87.8
6.78 ꢍ
6.69 ꢍ
6.78 ꢍ
4.88 ꢂ (4.8)
2.98–3.04 m
4.49 ꢁ-ꢋꢄkꢈ (9.0)
3.78 ꢃvꢈꢅꢋꢆppꢈꢂ
6.69 ꢍ
4.81 ꢂ (4.7)
2.96–3.02 m
4.45 ꢁ-ꢋꢄkꢈ (9.0)
62.7
72.2
62.6
72.3
9
9
1
2
3
4
5
6
7
8
9
9
ꢆ
b
′
3.73 ꢃvꢈꢅꢋꢆppꢈꢂ
128.2
106.2
149.0
136.4
149.0
106.2
89.4
134.7
106.6
154.0
135.8
154.0
106.6
89.1
′
6.71 ꢍ
6.73 ꢍ
′
′
′
′
′
′
′ꢆ
′b
6.71 ꢍ
4.65 (ꢍ)
6.73 ꢍ
4.67 ꢍ
92.9
76.3
93.1
76.2
4.09 ꢂ (9.6)
3.86 ꢃvꢈꢅꢋꢆppꢈꢂ
4.07 ꢂ (9.0)
3.84 ꢂ (9.3)
4.86 ꢂ (7.8)
3.43–3.47 m
3.37–3.42 m
3.37–3.42 m
3.15–3.20 m
3.64 ꢂꢂ (11.4, 4.8)
3.75 ꢃvꢈꢅꢋꢆppꢈꢂ
3.84 bꢅꢍ
105.3
75.7
77.8
71.3
78.4
62.6
4.81 ꢂ (7.6)
3.40–3.45 m
3.33–3.39 m
3.33–3.39 m
3.10–3.17 m
3.60 ꢂꢂ (12.0, 4.8)
3.71 ꢃvꢈꢅꢋꢆppꢈꢂ
3.82 ꢍ
105.4
75.8
77.9
71.3
78.4
62.6
+
+
+
+
57.1*
57.0*
56.8*
56.7*
57.1
57.0
56.8
56.8
3.84 bꢅꢍ
3.80 ꢍ
nꢃꢁꢈ: aꢋꢋ ꢆꢍꢍꢄgꢇmꢈꢇꢁꢍ wꢈꢅꢈ bꢆꢍꢈꢂ ꢃꢇ cosY, hsQc, hMBc ꢆꢇꢂ noesY ꢈxpꢈꢅꢄmꢈꢇꢁꢍ. *, +tꢀꢈ vꢆꢋꢊꢈꢍ mꢆy bꢈ ꢈxꢌꢀꢆꢇgꢈꢂ wꢄꢁꢀ
ꢈꢆꢌꢀ ꢃꢁꢀꢈꢅ.
Figure 2. sꢈꢋꢈꢌꢁꢈꢂ hMBc ꢆꢇꢂ noesY ꢌꢃꢅꢅꢈꢋꢆꢁꢄꢃꢇꢍ ꢃꢉ ꢌꢃmpꢃꢊꢇꢂꢍ 1 ꢆꢇꢂ 2.
C-4′ (Δδ 0.6 ppm) and the downfield shif s for C-1′ (Δδ 6.5 ppm), C-3′/C-5′ (Δδ 5.0 ppm),
1
3
and C-4 (Δδ 0.6 ppm) were observed in the C NMR spectrum of 2, suggesting that
the difference between 1 and 2 might be the location of the glucose moiety. Further
analysis of HMBC spectrum led to the confirmation of the location of the glucose at C-4′

4
Z.ꢀQ. ZꢁAꢂ ꢃT AL.
(
Figure2)bytheobservationofacorrelationbetweentheanomericprotonsignaloftheglucose
(
δ 4.81, d, J = 7.6 Hz) and C-4′ (δ 135.8). e ꢀ configuration of the glucose was assigned from
biogenetic consideration [5]. e specific rotation and the CD data for 2 were in agreement
with those of 1, which suggested that they possessed the same absolute configuration.

us, ussuriensislignan B was characterized as (–)-7S,7′R,8R,8′S-3,3′,5,5′-tetramethoxy-
4
,8′-dihydroxy-7,9′:7′,9-diepoxylignan-4′-O-β-ꢀ-glucopyranoside.
In addition to the two new compounds, seventeen known compounds: syringaresi-
nol-β-ꢀ-glucoside (3) [8], 4-(3′,4′-dihydroxyphenyl)butan-2-one-3′-O-β-ꢀ-glucopyranoside
(
4) [9–11], vanillyl alcohol-4-O-β-ꢀ-glucopyranoside (5) [12], 3-O-(β-ꢀ-glucopyranosyl)-1-
3′,5′-dimethoxy-4′-hydroxyphenyl)-1-propanone (6) [13], arbutin (7) [14], chlorogenic acid
(
methyl ester (8) [15], isorhamnetin-3-O-β-ꢀ-glucoside (9) [16], isorhamnetin-3-O-rutino-
side (10) [17], rutin (11) [18], quercetin 3-O-sophoroside (12) [19], n-butyl-β-ꢀ-fructopyra-
noside (13) [17], ethyl-β-ꢀ-fructopyranoside (14) [20], 1,5-dimethyl citrate (15) [21],
5
,5′-oxy(bismethylene)-2-furaldehyde (16) [22], ursolic acid (17) [23], oleanolic acid (18),
[
23] and daucosterol (19), were identified by comparing their spectroscopic data with lit-
erature data or co-TLC with authentic samples.
Compounds 7–12 exhibited potent antioxidant activity in both DPPH free radical-scav-
enging assay and ferric reducing power assay.
3
. Experimental
3
.1. General experimental procedures
Optical rotations were measured on a Rudolph Research Analytical Autopol II automatic
polarimeter (Rudolph, Flanders, NJ, U.S.A). e UV spectra were measured using an Agilent
Cary 60 UV–visible spectrophotometer (Agilent, Santa Clara, CA, U.S.A). e IR spectra
were recorded on a Bruker Tensor 27 spectrometer (Bruker, Karlsruhe, Germany). CD meas-
urements were carried out on MOS-500 spectrometer (BioLogic Science Instruments, Claix,
France). NMR spectra were collected on a Bruker AV-600 spectrometer (Bruker, Karlsruhe,
Germany) with TMS as internal standard. HR-ESI-MS were recorded on a Bruker micrO-
TOF-Q II mass spectrometer (Agilent, Bruker, U.S.A). Silica gel (100–200 and 200–300 mesh,
Qingdao Haiyang Chemical Co., Ltd. Qingdao, China), D101 macroporous resin (Tianjin
Haiguang Chemical Co., Ltd. Tianjin, China), ODS silica gel (LiChroprep RP-18, 40–63 μm,
Merck KGaA, Darmstadt, Germany), polyamide (60–100 mesh, Zhejiang Sijiashenghua
Plastic Co. Ltd., Zhejiang, China), and Sephadex LH-20 (Amersham Pharmacia Biotech
AB) were used for column chromatography. in Layer Chromatograghy (TLC) was car-
ried out using precoated plates with GF254 silica gel (Qingdao Haiyang Chemical Co.,
Ltd.). Preparative HPLC was performed using ODS columns (Agilent ZORBAX SB-C18,
2
1.2 mm × 250 mm, 7 μm). All other chemicals and reagents were of analytical grade and
purchased locally.
3
.2. Plant material
e fresh fruits of Pyrus ussuriensis were collected in October 2013 in Jixian County,
Tianjin, China, and identified by one of the authors (Prof. Tianxiang Li). A voucher


JꢂꢄꢅꢆAL ꢂF ASꢇAꢆ ꢆATꢄꢅAL PꢅꢂꢈꢄꢉTS ꢅꢃSꢃAꢅꢉꢁ
5
specimen (S201310002) has been deposited in the Laboratory of Natural Products, School
of Pharmaceutical Science and Technology, Tianjin University, China.
3
.3. Extraction and isolation
e fresh fruits of P. ussuriensis (16 kg) were washed and cut into small pieces, immersed

with 95% EtOH (25 L, for a week), and then refluxed with 95% (40 L, twice and each for 2 h)
and 60% (20 L, once for 2 h) EtOH (v/v). e ethanol extract (1.28 kg) was evaporated and
suspended in distilled water (4 L), and then partitioned with petroleum ether (60–90 °C),
EtOAc and n-BuOH successively. e n-BuOH extract was concentrated in vacuo, and
further separated on D101 macroporous resin column chromatography eluting with an
increasing concentration of EtOH (0, 50, 95%) in H O.
2

e ethyl acetate extract (76 g) was subjected to silica gel column (66 × 9.6 cm, 100–200
mesh) chromatography (petroleum ether–EtOAc, 8:2, 5:5, EtOAc–MeOH, 100:0, 9:1, 8:2,
5
:5, 500 ml each) to obtain six fractions (A–F). Fr. B (100 mg) was subjected to ODS col-
umn chromatography (MeOH–H O, 75:25, 80:20) to afford 17 (40 mg) and 18 (20 mg).
2
Fraction C (8.5 g) was loaded onto a polyamide column (20 × 6 cm) and eluted with
CH Cl –MeOH (96:4, 94:6, 9:1, 100 ml each) to obtain 60 subfractions. Subfrs. C11–C13
2
2
were recrystallized from MeOH to yield 19 (100 mg). Subfrs. C30–C51 were combined and
applied to polyamide column (24 × 2.6 cm) chromatography (CH Cl –MeOH, 96:4, 94:6)
2
2
and then further purified by Sephadex LH-20 column chromatography (MeOH) to afford 8
(
20 mg). Subfrs. C52–C63 (4 g) were combined on the basis of TLC analysis and subjected
to polyamide column (24 × 4 cm, 14.3 × 2.5 cm, stepwise) chromatography twice, eluting
with CH Cl –MeOH (96:4, 92:8). Compounds 7 (11 mg) and 9 (18 mg) were furnished
2
2
by further Sephadex LH-20 column chromatography. Fraction D (4.6 g) was loaded on
a silica gel column (27.3 × 4.6 cm, 200–300 mesh) using petroleum ether–Me CO (8:2,
2
7
1
4:26) as eluent and the resultant main fractions were recrystallized from MeOH to obtain
5 (103.7 mg). Fraction E (8 g) was loaded on a silica gel column (25 × 7.8 cm, 100–200
mesh) and eluted with EtOAc–MeOH (100:0, 94:6, 92:8, 80:20, 500 ml each) to obtain 76
subfractions. Subfrs. E33–E37 afforded 10 (12 mg) by further purification with Sephadex
LH-20 column chromatography (MeOH). Subfrs. E44–E58 were subjected to silica gel
column (21 × 2 cm, 200–300 mesh) chromatography (CH Cl –MeOH, 9:1, 8:2), polyamide
2
2
column (18.5 × 5 cm) chromatography (CH Cl –MeOH, 88:12, 86:14, 84:16, 8:2), and ODS
2
2
column chromatography (MeOH–H O, 25:75, 30:70, 40:60, 45:55), successively to yield 11
2
(
11.8 mg) and 12 (10.2 mg).

e 50% EtOH eluate (19 g) was chromatographed over a silica gel column (30 × 7 cm,
1
5
00–200 mesh) with a step gradient from ethyl acetate to methanol (100:0, 9:1, 8:2, 6:4, 3:7,
00 ml each) to produce seven fractions (A–G). Fr. A (174.1 mg) was sub-fractionated on a
silica gel column (15 × 2.7 cm, 200–300 mesh) with CH Cl –MeOH (99:1, 96:4, 20 ml each)
2
2
as eluent to give 34 subfractions. Subfrs. A24–A26 were chromatographed on a Sephadex
LH-20 column (MeOH) and purified by preparative TLC to yield 16 (2 mg). Fr. B (611 mg)
gave 4 (1 mg) by silica gel column (18 × 2.5 cm, 200–300 mesh) chromatography (CH Cl –
2
2
MeOH, 96:4) and Sephadex LH-20 column chromatography (MeOH). Separation of fr. C by
silica gel column (25 × 3 cm, 200–300 mesh) chromatography (CH Cl –MeOH, 94:6, 9:1, 8:2,
2
2
5
0 ml each) afforded 23 fractions, and 13 (63 mg) was furnished by further Sephadex LH-20
column chromatography from subfrs. C11–C15. Fr. D (686.4 mg) was subjected to silica gel

6
Z.ꢀQ. ZꢁAꢂ ꢃT AL.
column (16.7 × 2.6 cm, 200–300 mesh) chromatography (CH Cl –MeOH, 92:8, 9:1, 20 ml
2
2
each) and 14 (33 mg) was yielded by further Sephadex LH-20 column chromatography.
Fr. E (1.09 g) was chromatographed on a silica gel column (23.6 × 2.5 cm, 200–300 mesh)
eluted with CH Cl –MeOH (94:6, 8:2, 30 ml each), and subfrs. E14–E16 afforded 3 (3 mg).
2
2
Subfrs. E30–E41 (300 mg) were purified by preparative HPLC (29% MeOH–H O, flow rate
2
4
4
2
ml/min) af er Sephadex LH-20 column chromatography (MeOH) to give 5 (11.3 mg, tR
.1 min) and 6 (5 mg, t 6.0 min). Fr. F (3.4 g) was subjected to silica gel column (20 × 5 cm,
R
00–300 mesh) chromatography with isocratic elution of CH Cl –MeOH (84:16), and subfr.
2
2
F3 yielded 1 (5.7 mg, t 39.9 min) and 2 (2.7 mg, t 44.6 min) by preparative HPLC (30%
R
R
MeOH–H O, ꢁow rate 4 ml/min).
2
3.3.1. Ussuriensislignan A (1) 20
White amorphous powder; [ꢀ] −5.26 (c 0.57, MeOH); IR ν (KBr): 3417, 2954, 2838,
D
max
−
1
1
647, 1050, 1019, 684 cm ; UV (MeOH) λ (log ε) 210 (5.42), 272 (3.12) nm; CD (MeOH)
max
1
13
Δε242nm −0.524, Δε217nm++0.867; H and C NMR spectroscopic data, see Table 1; HRESIMS:
m/z 619.1990 [M+Na] (calcd for C H O Na, 619.1997).
2
8
36 14
3
.3.2. Ussuriensislignan B (2) 20
White amorphous powder; [ꢀ] −3.70 (c 0.27, MeOH); IR ν (KBr): 3453, 1642, 1019,
D
max
−
1
6
89 cm ; UV (MeOH) λ (log ε) 210 (5.74), 274 (4.46) nm; CD (MeOH) Δε245nm −0.103,
max
13
1
Δε214nm +0.152; H and C NMR spectroscopic data, see Table 1; HRESIMS: m/z 595.2027
−
[
M−H] (calcd for C H O , 595.2021).
2
8
35 14
3
.3.3. TLC acid hydrolysis of compounds 1 and 2
A methanol solution of compounds 1 and 2 was spotted on the TLC plate, respectively, and
fumigated by hydrochloric acid vapor at 70 °C for 20 min in the sealed container. en
the plate was removed from the hydrochloric acid vapor. Af er the plate was air-dried, the
authentic sugars were applied to the same plate. en, the plate was developed by EtOAc–
MeOH–H O–HAc (4:1:1:2.5), giving yellow spots at R 0.5 for glucose. 5% H SO in EtOH
2
f
2
4
was employed as a spray for visualization.
Acknowledgment
We are grateful to Prof. Benjamin Clark for polishing the language.
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding

is work was financially supported by Technological Project of Inner Mongolia Autonomous Region
[
NK 20150168].

JꢂꢄꢅꢆAL ꢂF ASꢇAꢆ ꢆATꢄꢅAL PꢅꢂꢈꢄꢉTS ꢅꢃSꢃAꢅꢉꢁ
7
References
[
[
[
[
1] J.Y. Cho, C.M. Kim, H.J. Lee, S.H. Lee, J.A. Cho, W.S. Kim, K.H. Park, and J.H. Moon, J. Agric.
Food Chem. 61, 4563 (2013).
2] L.J. Huang, W.Y. Gao, X. Li, W.S. Zhao, L.Q. Huang, and C.X. Liu, J. Agric. Food Chem. 58,
8983 (2010).
3] Y.G. Lee, J.Y. Cho, C.M. Kim, S.H. Lee, W.S. Kim, T.I. Jeon, K.H. Park, and J.H. Moon, Food
Sci. Biotechnol. 22, 803 (2013).
4] A.L. Piccinelli, S. Arana, A. Caceres, R.E.V. Bianca, R. Sorrentino, and L. Rastrelli, J. Nat. Prod.
6
7, 1135 (2004).
[
5] F. Yang, Y.F. Su, Y.P. Bi, J. Xu, Z.Q. Zhu, G.Z. Tu, and X.M. Gao, Helv. Chim. Acta 93, 536 (2010).
6] T.N. Vo, P.L. Nguyen, L.T. Tuong, L.M. Pratt, P.N. Vo, K.P.P. Nguyen, and N.S. Nguyen, Chem.
Pharm. Bull. 60, 1125 (2012).
[
[
[
[
7] H. Tsukamoto, S. Hisada, and S. Nishibe, Chem. Pharm. Bull. 32, 4482 (1984).
8] A.A. Shahat, N.S. Abdel-Azim, L. Pieters, and A.J. Vlietinck, Fitoterapia 75, 771 (2004).
9] K. Shimoda, T. Harada, H. Hamada, N. Nakajima, and H. Hamada, Phytochemistry 68, 487
(
2007).
[
[
[
10] A. Pabst, D. Barron, J. Adda, and P. Schreier, Phytochemistry 29, 3853 (1990).
11] I. Kouno, M. Inoue, Y. Onizuka, T. Fujisaki, and N. Kawano, Phytochemistry 27, 611 (1988).
12] H. Kanho, S. Yaoya, N. Kawahara, T. Nakane, Y. Takase, K. Masuda, and M. Kuroyanagi, Chem.
Pharm. Bull. 53, 361 (2005).
[
[
[
13] D.M. Zhang, Y. Li, and S.S. Geng, Nat. Prod. Res. Dev. 16, 496 (2004).
14] J. Chen, J.J. Chen, and K. Gao, Chem. Nat. Compd. 49, 1129 (2014).
15] V.D. Kancheva, L. Saso, S.E. Angelova, M.C. Foti, A. Slavova-Kasakova, C. Daquino, V. Enchev,
O. Firuzi, and J. Nechev, Biochimie 94, 403 (2012).
[
16] A.N. Shikov, M. Kundracikova, T.L. Palama, O.N. Pozharitskaya, V.M. Kosman, V.G. Makarov,
B. Galambosi, H.J. Kim, Y.P. Jang, Y.H. Choi, and R. Verpoorte, Phytochem. Lett. 3, 45 (2010).
17] S.Y. Lee, S.U. Choi, J.H. Lee, D.U. Lee, and K.R. Lee, Arch. Pharm. Res. 33, 515 (2010).
18] B.T.T. Luyen, B.H. Tai, N.P. ao, K.J. Eun, J.Y. Cha, M.J. Xin, Y.M. Lee, and Y.H. Kim, Bioorg.
Med. Chem. Lett. 24, 1895 (2014).
[
[
[
[
[
[
[
19] R. Nørbæk and T. Kondo, Phytochemistry 51, 1113 (1999).
20] J.T. Liu, Y. Tang, K.G. Wu, C.F. Bi, and Q. Cui, Carbohydr. Res. 350, 20 (2012).
21] R.J. Ansell, J.E. Meegan, S.A. Barrett, and S.L. Warrinner, Dalton Trans. 8, 1460 (2009).
22] O. Casanova, S. Iborra, and A. Corma, J. Catal. 275, 236 (2010).
23] W. Seebacher, N. Simic, R. Weis, R. Saf, and O. Kunert, Magn. Reson. Chem. 41, 636 (2003).