全 文 ：书天然产物研究与开发 Nat Prod Res Dev 2012，24:861-865
文章编号:1001-6880(2012)07-0861-05
Received March 31，2012 Accepted May 17，2012
Foundation Item:This study was supported by the National Natural Sci-
ence Foundation of China (No． 20962013)
* Corresponding author Tel:86-871-5920671;E-mail:lihaizhou@ yninfo．
com
菝葜中三个新的二氢山萘酚酰化糖苷
郝 倩，姚 尧，李晓蕾，李蓉涛，李海舟*
昆明理工大学 生命与科学技术学院，昆明 650500
摘 要:从菝葜根茎中共分离得到 3 个新的二氢黄酮醇酰化糖苷(1 ～ 3)和 1 个已知的二氢黄酮醇苷(4) ，分别
被鉴定为:(2R，3R)-二氢山萘酚 3-氧-(6″-氧-乙酰基)-β-D-葡萄糖苷 (1) ，(2R，3R)-二氢山萘酚 3-氧-(2″-氧-乙
酰基)-β-D-葡萄糖苷 (2) ，(2R，3R)-二氢山萘酚 3-氧-(3″-氧-乙酰基)-β-D-葡萄糖苷 (3)及(2R，3R)-二氢山萘
酚 3-氧-β-D-葡萄糖苷 (4)。其结构通过 MS，1D NMR和 2D NMR等光谱分析和化学方法确定。研究结果表明，
二氢黄酮醇的酰化糖苷可能是菝葜属的特征成分。
关键词:菝葜属;二氢山萘酚酰化糖苷;二氢黄酮醇苷;黄酮类
中图分类号:R284. 1 文献标识码:A
Three New Dihydrokaempferol Acetylated Glycosides from Smilax china L．
HAO Qian，YAO Yao，LI Xiao-lei，LI Rong-tao，LI Hai-zhou*
The Faculty of Life Science and Technology，Kunming University of Science and Technology，Kunming 650500，China
Abstract:Three new flavanonol acetylated glycosides (1-3) ，together with a known flavanonol glycoside (4) ，were isola-
ted from the rhizomes of Smilax china L． Their structures were elucidated on the basis of MS，1D and 2D NMR spectro-
scopic analysis and chemical evidence． The new compounds were characterized as (2R，3R)- dihydrokaempferol 3-O-
(6-O-acetyl)-β-D-glucopyranoside (1) ，(2R，3R)-dihydrokaempferol 3-O-(2-O-acetyl)-β-D-glucopyranoside (2) ，
and (2R，3R)-dihydrokaempferol 3-O-(3-O-acetyl)-β-D-glucopyranoside (3)． The results indicate that flavanonol
acetylated glycosides probably can serve as typical markers of the genus Smilax．
Key words:Smilax china;dihydrokaempferol acetylated glycosides;flavanonol glycosides;flavonoids
Introduction
The genus Smilax(family Liliaceae;about 350 species)
plants are distributed widely in tropical and temperate
regions throughout the world，especially in East Asia
and North America． Many of them have long been used
as medicinal herbs［1-8］． Previous phytochemical investi-
gations on this genus revealed the presence of steroid
saponins and flavonoid glycosides as the major chemi-
cal constituents． The rhizomes of Smilax china L．，
known as“Ba Qia”or“Jin Gang Teng”in China，is
used in traditional Chinese medicine (TCM) for the
treatment of diuretic，rheumatic arthritic，detoxication，
lumbago，gout，tumor and inflammatory diseases［9］．
Pharmacological research showed that S． china has anti-
inflammatory，anti-nociceptive，free radical scavenging，
neuroprotective，antioxidant，and antimicrobial activi-
ties［9-12］． Previous phytochemical studies on this plant
led to the isolation of several steroid saponins，inclu-
ding smilaxin，prosapogenin A of dioscin，gracillin，di-
oscin，pseudoprotodioscin，methygracillin，methylproto-
dioscin and smilaxchinosides A-D［13，14］．
The present work focuses on the isolation and structural
elucidation of flavonoids from the n-BuOH extract of S．
china． Four compounds were isolated and indentified．
Three are new flavanonol acetylated glycosides (1-3)
and one known flavanonol glycoside (4)．
Fig. 1 Compounds isolated from Smilax china L．
Experimental
General
Optical rotations were measured on HORIBA SEPA-
300 high-sensitive polarimeter． 1D and 2D NMR spec-
tra were recorded in DMSO-d6using Bruker DRX-500
operating at 500 MHz for 1H，125 MHz for 13C，respec-
tively． ESI-MS and HR-ESI-MS were taken on an API
Qstar Pulsar-1 mass spectrometer． Column chromatogra-
phy (CC)was carried on silica gel (200-300 mesh，
100-200 mesh，Qingdao Marine Chemical Factory，Chi-
na) ，Sephadex LH-20 (25-100 μm，Pharmacia Fine
Chemical Co．，Ltd．，USA) ，and ODS (YMC* Gel
ODS-A，50 μm，YMC Co．，Ltd．，Japan)． Fractions
were monitored by TLC precoated plates (Si gel GF254，
Qingdao Marine Chemical Factory，China) ，and spots
were detected by spraying with 2% FeCl3-EtOH or
10% H2SO4-EtOH reagent followed by heating． Semi-
preparative HPLC was performed on an Agilent 1200
liquid chromatographic system (Agilent Technologies
Co．，Ltd．，USA)with Zorbax SB-C18(Agilent，9． 4 mm
× 250 mm)column． All chemical solvents used for i-
solation were of analytical grade or higher．
Plant Material
The rhizomes of S． china were purchased from Guilin
City，Guanxi Province，China，in March，2010，and were
identified by Dr． Hai-Zhou Li． A voucher specimen
(KMUST-B-2010030101)was deposited at the Labo-
ratory of Phytochemistry，the Faculty of Life Science
and Technology，Kunming University of Science and
Technology．
Extraction and isolation
The air-dried and powdered rhizomes of S． china(220
g)were ultrasonic extracted with 70% aq． EtOH (3 ×
1 L，30 min，each)at room temperature． The extracting
solution was evaporated under reduced pressure to yield
an extract，which was suspended in H2O and then par-
titioned with petroleum ether and n-BuOH，respective-
ly．
The n-BuOH fraction (28 g)was separated over Seph-
adex LH-20 (MeOH /H2O 0∶ 1，3∶ 7，6∶ 4，9∶ 1，and 1∶
0)to afford four fractions (I-IV)． Fraction II (1． 15 g，
MeOH/H2O 3∶ 7)was separated by ODS medium-pres-
sure liquid chromatography (MPLC) (MeOH /H2O 3
∶ 7-6∶ 4) ，followed by MPLC over silica gel (CHCl3 /
MeOH 8 ∶ 1) to yield compounds 1 (11． 1 mg) ，2
(14. 3 mg)and 3 (6． 3 mg) ，successively． Fraction III
(5． 48 g，MeOH /H2O 4∶ 6)was subjected to semi-pre-
parative HPLC on Zorbax SB-C18(MeOH /H2O 4 ∶ 6，
flowing rate 3 mL /min)to yield compound 4 (10 mg，
12. 5min)．
Acid hydrolysis
The Fraction II (100 mg)was heated at 95 ℃ with 2
M HCl (15 mL)for 12 h，then，neutralized with NaH-
CO3solution，the reaction mixture was extracted with
CHCl3(3 × 15 mL)． The organic layer was evaporated
and subjected to silica gel PTLC using CHCl3 /MeOH /
H2O (8∶ 2∶ 0． 2)as eluent to yield flavanonol aglycone
5(15． 2 mg) ，which identified as (2R，3R)dihydro-
kaempferol by comparing 1H NMR spectroscopic data
and optical rotation with the reference［15］． The H2O
layer was then concentrated and submitted to silica gel
PTLC (CHCl3 /MeOH /H2O 7∶ 3∶ 0． 5)to yield the D-
glucose (3． 6 mg) ，which was identified by co-TLC
comparison with the authentic sample and optical rota-
tion［α］25D = + 58． 3 (c = 0． 1，MeOH)．
Results and Discussion
The structure of the known compound 4 was character-
ized as (2R，3R)-dihydrokaempferol 3-O-β-D-glucopy-
ranoside ［16］ on the basis of NMR spectroscopic data
and the value of optical rotation． The spectroscopic fea-
tures of the new compounds 1-3 were very similar to
those of 4． The FeCl3 reaction，Molish reaction and
NaBH4 reaction were positive，suggesting they were fla-
vanonol glycosides．
Compound 1 was isolated as a yellow amorphous pow-
der． Its molecular formula was deduced to be C23H24O12
on the basis of the negative HR-ESI-MS (m/z
491. 1195，［M-H］-)and the comprehensive analysis
of NMR data． The1H NMR spectrum of 1 showed the
two doublets for four aromatic protons at δ7． 19 and
6. 72(each 2H，J = 8． 6 Hz)assigned to H-2 /6 and
H-3 /5 of the favanonol，respectively． A pair of dou-
blet aromatic proton signals at δ 5． 89 and 5． 86 (each
1H，d，J = 2． 0 Hz)were assigned to H-6 and H-8． And
268 Nat Prod Res Dev Vol. 24
the coupling constants between H-2 and H-3 at δ 5． 54
(1H，d，J = 7． 2 Hz)and 4． 74 (1H，d，J = 7． 2 Hz)
showed its 2，3-trans-configuration，and the value of op-
tical rotation of the aglycone after acid hydrolysis sug-
gested 1 was a (2R，3R)-dihydrokaempferol deriva-
tive． In addition，the anomeric proton signals［δ4． 20
(1H，d，J = 8． 0 Hz) ］，an acetyl methyl signal at δ
1. 97 (3H，s)were also observed，together with signals
between δ 3． 00 and 3． 40 attributed to the remaining
protons of sugar residues． Analysis of the 13C NMR
spectrum revealed that 1 contains 23 carbons，including
one acetyl moiety (δC 20． 6 and 170． 3) ，one glucosyl
unit (δC 60． 6，69． 7，73． 3，73． 9，76． 3，and 100． 9) ，
six tri-substitute olefinic carbons (δC 95． 1，96． 0，115．
2 × 2 and 128． 7 × 2) ，and seven quaternary aromatic
carbons (δC 101． 1，126． 1，157． 7，161． 7，163． 5，167． 3 and
192． 8)．
Careful analysis of the 1H and 13C NMR (Table 1)
spectral data indicated 1 to be a flavanonol glycoside
similar to the known (2R，3R)-dihydrokaempferol 3-O-
β-D-glucopyranoside (4)［16］，except for the presence
of an acetyl moiety． HMBC correlation of the proton on
glucose at δ 4． 16 (H-6)with the carbonyl carbon
signal at δ 170． 3 (MeCO-6)suggested that the
acetyloxyl moiety was located at C-6． On the basis of
the above spectroscopic evidence，the structure of 1 was
identified as (2R，3R)-dihydrokaempferol 3-O-(6-O-
acetyl)-β-D-glucopyranoside．
Table 1 1H，13C-NMR data of compounds 1-4 in DMSO-d6(δ in ppm，J in Hz)
No．
1 2 3 4
δH δC δH δC δH δC δH δC
2 5． 54 (1H，d，7． 2) 80． 8 5． 35 (1H，d，9． 6) 80． 9 5． 46 (1H，d，8． 2) 80． 8 5． 45 (1H，d，8． 0) 81． 0
3 4． 74 (1H，d，7． 2) 75． 9 4． 95 (1H，d，9． 6) 75． 8 5． 01 (1H，d，8． 2) 75． 0 4． 94 (1H，d，8． 2) 75． 4
4 192． 8 193． 1 193． 0 193． 0
5 163． 5 163． 4 163． 4 163． 5
6 5． 89 (1H，d，2． 0) 96． 0 5． 89 (1H，d，2． 0) 96． 1 5． 89 (1H，d，2． 4) 95． 9 5． 86 (1H，d，2． 0) 96． 2
7 167． 3 166． 9 167． 1 167． 0
8 5． 86 (1H，d，2． 0) 95． 1 5． 87 (1H，d，2． 0) 94． 9 5． 88 (1H，d，2． 4) 94． 9 5． 86 (1H，d，2． 0) 95． 0
9 161． 7 162． 0 161． 8 162． 1
10 101． 1 101． 0 101． 0 100． 9
1 126． 1 126． 2 126． 1 126． 1
2，6 7． 19 (2H，d，8． 6) 128． 7 7． 28 (2H，d，8． 6) 129． 3 7． 26 (2H，d，8． 5) 128． 9 7． 26 (2H，d，8． 4) 129． 0
3，5 6． 72 (2H，d，8． 6) 115． 2 6． 79 (2H，d，8． 6) 115． 1 6． 75 (2H，d，8． 5) 115． 1 6． 75 (2H，d，8． 4) 115． 2
4 157． 7 157． 9 157． 7 157． 8
5-OH 11． 61 (1H，s) 11． 74 (1H，s) 11． 69 (1H，s) 11． 70 (1H，s)
Glc-1 4． 20 (1H，d，8． 0) 100． 9 3． 88 (1H，d，8． 0) 98． 7 4． 01 (1H，d，7． 7) 100． 4 3． 97 (1H，d，6． 4) 101． 0
2 3． 40 － 3． 00 (1H，m) 73． 3 4． 42 (1H，t，10． 5) 73． 4 3． 18 (1H，t，8． 0) 72． 4 2． 90 (1H，m) 73． 5
3 3． 01 (1H，d，8． 1) 76． 3 3． 03 (1H，t，8． 2) 74． 2 3． 37 (1H，t，8． 3) 77． 6 3． 09 (1H，m) 76． 7
4 3． 00 (1H，d，8． 0) 69． 7 3． 02 (1H，t，8． 2) 70． 0 3． 09 (1H，t，8． 2) 71． 2 2． 94 (1H，m) 70． 2
5 3． 40 ～ 3． 00(1H，m) 73． 9 2． 93 (1H，m) 77． 6 3． 06 (1H，m) 76． 9 3． 09 (1H，m) 77． 3
6a 4． 16 (1H，d，11． 8) 60． 6 3． 62 (1H，d，10． 3) 60． 9 3． 49 (1H，d，10． 9) 60． 7 3． 66 (1H，m) 61． 2
6b 3． 96(1H，dd，11． 8，4． 0) 3． 39(1H，dd，10． 3，3． 8) 3． 28 (1H，dd，10． 3，3． 9) 3． 28 (1H，m)
CH3CO 170． 3 169． 2 169． 7
CH3CO 1． 97 (3H，s) 20． 6 1． 24 (3H，s) 20． 9 1． 22 (3H，s) 21． 0
The molecular formula of compounds 2 and 3 were as- signed as C23 H24 O12，the same as that of 1，as deter-
368Vol. 24 HAO Qian，et al:Three New Dihydrokaempferol Acetylated Glycosides from Smilax china L．
mined from their molecular ion ［M-H］-peaks at m/z
491． 1189 and 491． 1184 in HR-ESI-MS，respectively．
Their 1H NMR spectra indicated that they shared a pair
of doublet aromatic proton signals around δ 5． 8，A2B2-
type aromatic proton signals at δ 6． 7-7． 3，and glucose
sugar moieties． From their 1H NMR spectra，2 and 3
were deduced to be as 2，3-trans flavanonol glucosides
from their H-2 coupling constants［2:δ 5． 35 (1 H，d，
J = 9． 6 Hz) ，3:δ 5． 46 (1 H，d，J = 8． 2 Hz) ］． The
HMBC spectra indicated that these compounds bore a
glucose moiety at their C-3 position，as determined by
the H-Clong range correlations of H-1［2:δ 3． 88 (1
H，d，J = 8． 0 Hz) ，3:δ 4． 01 (1 H，d，J = 7． 7 Hz) ］to
C-3 (2:δ 75． 8，3:δ 75． 0)． Additionally，the bonding
positions of their acetyl moieties were confirmed from
the HMBC correlations between their glucose proton
signals［2:δ 4． 42 (1 H，t，J = 10． 5 Hz，H-2) ，3:δ
3. 37 (1 H，t，J = 8． 3 Hz，H-3) ］and carbonyl car-
bon signals ［2:δ 169． 2 (MeCO-2) ，3:δ 169． 7
(MeCO-3) ］． From these results，their structures were
determined to be (2R，3R)-dihydrokaempferol 3-O-
(2-O-acetyl)-β-D-glucopyranoside (2) and (2R，
3R)-dihydrokaempferol 3-O-(3-O-acetyl)-β-D-glu-
copyranoside (3) ，respectively．
In this study，three new flavanonol acetylated glycosides
(1-3) ，together with a known flavanonolglycoside (4) ，
were isolated from the rhizomes of S． china． In addi-
tion，three dihydroflavonol acetylated glycosides，(2R，
3R)-2-acetyl astilbin，(2R，3R)-3-acetyl astilbin，
(2R，3R)-4-acetyl astilbin were reported from Smilax
corbularia［17］． These results suggested that the di-
hydroflavonoid acetylated glycosides probably can serve
as typical markers of the genus Smilax．
Compound 1:amorphous powder，［α］16． 8D = -6． 8 (c
= 0． 79，MeOH)． 1H (500 MHz)and 13C (125 MHz)
NMR data (DMSO-d6) :see Table 1． ESI-MS (neg．) :
m/z 491［M-H］-，HR-ESI-MS (neg．) :m/z 491． 1195
［M-H］-(C23H23O
-
12，calc． 491． 1189)．
Compound 2:amorphous powder，［α］17． 2D = -10． 5 (c
= 0． 33，MeOH)． 1H (500 MHz)and 13C (125 MHz)
NMR data (DMSO-d6) :see Table 1． ESI-MS (neg．) :
m/z 491［M-H］-，HR-ESI-MS (neg．) :m/z 491． 1189
［M-H］-(C23H23O
-
12，calc． 491． 1189)．
Compound 3:amorphous powder，［α］16． 9D = -9． 11 (c
= 0． 30，MeOH)． 1H (500 MHz)and 13C (125 MHz)
NMR data (DMSO-d6) :see Table 1． ESI-MS (neg．) :
m/z 491［M-H］-，HR-ESI-MS (neg．) :m/z 491． 1184
［M-H］-(C23H23O12-，calc． 491． 1189)．
Compound 4:amorphous powder，［α］25D = + 35． 8 (c
= 0． 20，MeOH)． 1H (500 MHz)and 13C (125 MHz)
NMR data (DMSO-d6) :see Table 1． ESI-MS (neg．) :
m/z 449［M-H］-．
Compound 5:amorphous powder，［α］25D = + 30． 6 (c
= 0． 2，MeOH)． 1H (500 MHz)NMR data (DMSO-
d6) :11． 90 (1H，s，5-OH) ，10． 79 (1H，s，7-OH) ，
9. 40 (1H，s，4-OH) ，7． 28 (2H，d，J = 8． 5 Hz，H-2，
6) ，6． 78 (2H，d，J = 8． 5 Hz，H-3，5) ，5． 85 (1H，
d，J = 2． 0 Hz，H-6) ，5． 89 (1H，d，J = 2． 0 Hz，H-8) ，
5． 04 (1H，d，J = 11． 0 Hz，H-2) ，4． 65 (1H，d，J =
2. 0 Hz，H-3)．
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568Vol. 24 HAO Qian，et al:Three New Dihydrokaempferol Acetylated Glycosides from Smilax china L．