全 文 ：· 100 · 药学学报 Acta Pharmaceutica Sinica 2016, 51 (1): 100−104

s


A new caffeate compound from Nardostachys chinensis
CHEN Ying-peng1, 2, WANG Zhong-ping1, 2, ZHENG Hong-hong1, 2, XU Yan-tong1, 2,
ZHU Yan1, 2, ZHANG Peng1, 2, WU Hong-hua1, 2*
(1. Tianjin State Key Laboratory of Modern Chinese Medicine, Institute of Traditional Chinese Medicine, Tianjin University of
Traditional Chinese Medicine, Tianjin 300193, China; 2. Research and Development Center of Traditional Chinese Medicine,
Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin 300457, China)
Abstract: A new caffeate compound, (E)-erythro-syringylglyceryl caffeate (1), was isolated from the
roots and rhizomes of Nardostachys chinensis Batal., together with nine known phenolic compounds, including
(+)-licarin A (2), naringenin 4, 7-dimethyl ether (3), pinoresinol-4-O-β-D-glucoside (4), caraphenol A (5), Z-
miyabenol C (6), protocatechuic acid (7), caffeic acid (8), gallic acid (9) and vanillic acid (10). Their chemical
structures were elucidated on the basis of spectroscopic data and physicochemical properties. Furthermore, this
is the first report of compounds 2, 5 and 6 from Nardostachys genus.
Key words: Nardostachys chinensis; caffeate compound; chemical constituent; phenolic compound
CLC number: R284.2 Document code: A Article ID: 0513-4870 (2016) 01-0100-05
甘松中一个新的咖啡酸酯类化合物
陈应鹏 1, 2, 王忠平 1, 2, 郑红红 1, 2, 徐砚通 1, 2, 朱 彦 1, 2, 张 鹏 1, 2, 吴红华 1, 2*
(1. 天津中医药大学中医药研究院, 天津市现代中药重点实验室, 天津 300193;
2. 天津国际生物医药联合研究院中药新药研发中心, 天津 300457)

摘要: 从甘松 (Nardostachys chinensis Batal.) 中分离得到 1个新的咖啡酸酯类化合物和 9个已知酚类化合物,
通过物理化学性质和波谱学方法鉴定了它们的化学结构, 分别为 (E)-咖啡酸−赤式−紫丁香基甘油醇酯 (1)、(+)-
licarin A (2)、柚皮素-4, 7-二甲醚 (3)、(+)-松脂素-4-O-β-D-吡喃葡萄糖苷 (4)、caraphenol A (5)、Z-miyabenol C (6)、
原儿茶酸 (7)、咖啡酸 (8)、没食子酸 (9) 和香草酸 (10)。其中, 化合物 1为新化合物, 2、5和 6为首次从该属
植物中分离得到。
关键词: 甘松; 咖啡酸酯类化合物; 化学成分; 酚类化合物

Nardostachys chinensis Batal. (NCB), a member
of the genus Nardostachys (Valerianaceae), is distributed
mainly in Sichuan, Gansu, and Qinghai provinces in
China. The rhizome and root of NCB, also called
“Gansong or Gansongxiang”, was used as an herbal drug

Received 2015-05-25; Accepted 2015-07-08.
Project supported by the International Science & Technology Cooperation
Program of China (2013DFA31620).
*Corresponding author Tel / Fax: 86-22-59596163,
E-mail: wuhonghua2003@163.com
DOI: 10.16438/j.0513-4870.2015-0483
in traditional Chinese medicine for centuries to elicit
the stomachic and sedative effects[1]. “Gansong” is
now also a well-known medicinal spice used in Chinese
hotpot, especially numb and spicy hotpot originates in
Songpan County, Sichuan Province in China[2].
In order to exploit natural 5-hydroxytryptamine
transporter (SERT) regulators from traditional Chinese
medical herbs, we’ve promoted a systematical study on
the bioactive chemical constituents of NCB. In this
report, isolation and purification of the phenolic com-
pounds (Figure 1), recently isolated from NCB, as well
CHEN Ying-peng, et al: A new caffeate compound from Nardostachys chinensis · 101 ·



Figure 1 The chemical structures of compounds 1−10

as the structural elucidation of these compounds based
on their spectroscopic data and physicochemical prop-
erties was summarized.

Results and discussion
Compound 1 was obtained as a white amorphous
powder. The molecular formula of this compound was
assigned to be C20H22O9, as deduced from its quasi-
molecular ion peak at (−)-ESI-MS m/z 405.73 [M−H]−
and NMR spectroscopic data (Table 1).
In the 1H NMR spectrum of compound 1, the sig-
nals at δH 6.26 (1H, d, J = 16.2 Hz) and 7.46 (1H, d, J =
16.2 Hz) indicated the existence of a trans-form double
bond in 1; the signals at δH 7.00 (1H, dd, J = 8.4, 1.8
Hz), 6.76 (1H, d, J = 8.4 Hz) and 7.04 (1H, br s), and
the signals at δH 6.61 (2H, s) suggested respectively
the presences of a 1,3,4-trisubstituted and a 1,3,4,5-
tetrasubstituted benzene rings; and the signal at δH 3.73
(6H, s) indicated two magnetic equivalent methoxy
groups of 1. The 13C NMR spectrum of compound 1
confirmed the above deduction and showed another one
oxygenated methylene (δC 65.7) and two oxygenated
methines (δC 73.6, 72.9) in the more upfield area.
Further analysis of the HSQC spectrum disclosed that
δH 4.95 (1H, d, 4.8), 5.23 (1H, d, 4.2), 9.59 (1H, br s),
8.17 (1H, br s) and 9.15 (1H, br s) were five active
hydroxyl proton signals. The HMBC correlations
(Figure 2) from δH 7.46 (H-7) to δC 114.8 (C-2), 121.3

Table 1 NMR spectroscopic data for 1 (600 MHz for 1H, 150
MHz for 13C, DMSO-d6)
Position δH (J in Hz) δC
1 − 125.5
2 7.04 (1H, br s) 114.8
3 − 145.6
4 − 148.4
5 6.76 (1H, d, 8.4) 115.7
6 7.00 (1H, dd, 8.4, 1.8) 121.3
7 7.46 (1H, d, 16.2) 145.0
8 6.26 (1H, d, 16.2) 114.0
9 − 166.5
1 − 132.6
2 (6) 6.61 (2H, s) 104.2
3 (5) − 147.5
4 − 134.5
7 4.46 (1H, dd, 4.2, 4.8) 73.6
8 3.78 (1H, m) 72.9
9 4.00 (1H, dd, 10.8, 3.0) 65.7
3.84 (1H, dd, 10.8, 7.2)
3, 5-OMe 3.73 (6H, s) 55.9
4-OH 8.17 (1H, br s) −
7-OH 4.95 (1H, d, 4.8) −
8-OH 5.23 (1H, d, 4.2) −
ph-OH 9.59 (1H, br s) −
9.15 (1H, br s) −
· 102 · 药学学报 Acta Pharmaceutica Sinica 2016, 51 (1): 100−104


Figure 2 The main moieties and the key HMBC (H C)
correlations of compound 1

(C-6), and 166.5 (C-9), from δH 4.46 (H-7) to δC 132.6
(C-1), 104.2 (C-2/6) and 72.9 (C-8), and from δH 3.73
to δC 147.5, indicated the presences of (E)-caffeoyl and
syringylglyceryl groups in the structure of compound
1. As was expected, the two groups came to forming
an ester, confirmed by the HMBC correlation from
δH 3.84 (H-9) to δC 166.5 (C-9). Considering the
coupling constant between H-7 and H-8 was 4.3 Hz,
the configuration between H-7 and H-8 should prefer
an erythro-form[3, 4]. Thus, the structure of compound
1 was finally confirmed as (E)-erythro-syringylglyceryl
caffeate.
Considering that the optical rotation ([α] 20D −1.22
(c 0.60, MeOH) and CD spectra [λ (Δε): 203 (+7.32)
nm, c 0.53, MeOH] of 1 was observed not significant
and unstable, while the NMR spectra indicated there was
no threo-conformer mixed in it, therefore, compound
1 was deduced as a racemic mixture of two erythro-
isomers with contrast absolute configurations. So,
compound 1 was finally confirmed as a mixture of
(E)-7R, 8R-syringylglyceryl caffeate and (E)-7S, 8S-
syringylglyceryl caffeate.
Another nine known compounds were elucidated as
(+)-licarin A (2)[5], naringenin 4, 7-dimethyl ether (3)[6],
pinoresinol-4-O-β-D-glucoside (4)[7], caraphenol A (5)[8],
Z-miyabenol C (6)[9], protocatechuic acid (7)[10], caffeic
acid (8)[11], gallic acid (9)[8] and vanillic acid (10)[12] by
comparing their spectroscopic data (including 1D/2D
NMR, ORD and CD experiments) and physicochemical
properties with the reported data. Furthermore, the
relative configurations of the two resveratrol oligomers
5 and 6 were confirmed on the basis of their NOESY/
ROESY spectra (Figure 3 and 4) and ORD/CD properties.
Among these nine known compounds, compounds 2, 5
and 6 were isolated from Nardostachys genus for the
first time.

Experimental
General experimental procedures All reagents
were of HPLC or analytical grade. Column chromato-
graphy (CC) was performed over silica gel (SiO2; 200−
300 mesh), Sephadex LH-20, polyamide (80−100 mesh)
and D101 macroporous resin; TLC was conducted on
precoated silica gel plates GF254 (SiO2; 400−500 mesh).


Figure 3 The key HMBC (H C) and NOESY (H H) correlations of compound 5


Figure 4 The key HMBC (H C) and ROESY (H H) correlations of compound 6
CHEN Ying-peng, et al: A new caffeate compound from Nardostachys chinensis · 103 ·

Preparative HPLC was performed using an Agilent
Zorbax SB-C18 ODS column (21.2 mm × 250 mm, 7 μm)
at 10 mL·min−1. NMR spectra were recorded on Bruker
AV-III spectrometer using TMS as the internal standard.
UV and CD were recorded with a Jasco J-815 Circular
Dichroism (CD) spectropolarimeter. Specific optical
rotation values ([α] 20D ) were recorded with a Rudolph
AUTOPOL V polarimeter. IR spectra were obtained
on a Perkin Elmer Spectrum 65 FT-IR spectrometer.
ESI-MS were measured on a Waters Quattro Premier
XE mass spectrometer.
Plant material The dry roots and rhizomes of
Nardostachys chinensis Batal. were purchased from
Anhui Jiren Pharmacy Co., Ltd., P. R. China, in July,
2011. The plant material was authenticated by Prof.
Tian-xiang Li, Tianjin University of Traditional Chinese
Medicine, P. R. China. A voucher specimen (No.
B20604126) was deposited in the Traditional Chinese
Medicine Research and Development Center, Tianjin
University of Traditional Chinese Medicine.
Extraction and isolation The air-dried under-
ground parts (20 kg) of Nardostachys chinensis Batal.
were extracted successively with cold and hot 70%
EtOH. The 70% EtOH extract (3.4 kg) combined was
then evaporated under reduced pressure and distributed
in water before being partitioned with petroleum ether
(PE), ethyl acetate (EA), and n-butanol (BU) succes-
sively. The BU fraction, combined with the fraction
afforded by alcohol precipitation of the rest water solu-
tion, was chromatographed on D101 macroporous resin
column, gradiently eluted with EtOH-H2O (0∶100−
95∶5) to obtain 5 fractions. Among them, the 95%
EtOH fraction was added to the EA fraction.
The PE extract (320 g) was chromatographed
on silica gel column, gradiently eluted with PE-EA
(100∶0−0∶100) to obtain 22 fractions (PE.1−PE.22).
Fractions PE.6−9 were then subjected to silica gel
column gradiently eluted with PE-EA (100∶0−100∶30)
respectively, and afforded compounds 3 (11.0 mg) and
2 (7.0 mg).
The EA extract (1.2 kg) was separated into 15
fractions (EA.1−EA.15) on a normal-phase silica gel
column eluted with CHCl3−MeOH (from 100∶0 to
0∶100, v/v) solvent. Fraction EA.8 (38.8 g) was
further purified by subjecting to polyamide column,
Sephadex LH-20 column and HPLC chromatography to
yield compound 7 (MeOH−H2O = 30∶70, tR = 9.18 min,
350.0 mg) and 8 (MeOH−H2O = 22∶78, tR = 28.88 min,
256.0 mg). Fraction EA.9 (10.8 g) was separated
into seven subfractions (EA.9-1−EA.9-7) by polyamide
column chromatography with a stepwise gradient elution
of MeOH−H2O (0∶100−100∶0). Subfraction EA.9-2
(0.9 g) was subjected to preparative HPLC (MeOH−
H2O−HCOOH = 40∶60∶0.1) to yield compound 4
(tR = 20.11 min, 25.0 mg). Subfraction EA.9-4 and
EA.9-7 was then further isolated to yield compound 1
(tR = 60.30 min, 9.4 mg), compound 5 (tR = 40.35 min,
7.1 mg), compound 6 (tR = 20.11 min, 7.4 mg) and
compound 9 (240.0 mg).
The 30% EtOH fraction (190 g) was chroma-
tographed on D101 macroporous resin column, gradi-
ently eluted with MeOH−H2O (10∶90−100∶0) to
obtain 5 subfractions. The 10% MeOH subfraction
(20 g) was then further isolated by combined Sephadex
LH-20 (MeOH−H2O = 1∶1) column and preparative
HPLC (MeOH−H2O = 8∶92) chromatography method
to afford compound 10 (tR = 96.30 min, 20.0 mg).
(E)-erythro-Syringylglyceryl caffeate (1): white
amorphous powder; C20H22O9; [α] 20D −1.22 (c 0.60,
MeOH); UV (MeOH) λmax (logε): 209 (3.13), 331 (2.83)
nm; CD (c 0.53, MeOH) λ (Δε): 203 (+7.32) nm; IR
(KBr) νmax: 2 960, 2 927, 2 854, 1 695, 1 602, 1 516,
1 494, 1 453, 1 375, 1 326, 1 269, 1 184, 1 046, 1 024,
990, 907, 823, 759 cm−1; 1H and 13C NMR see Table 1;
(−)-ESI-MS m/z 405.73 [M−H]−.
Caraphenol A (5): yellow amorphous powder;
C42H28O9; [α] 20D +148.76 (c 0.51, MeOH); UV (MeOH)
λmax (logε): 210 (3.33), 267 (2.91), 287 (3.20), 308 (2.57),
329 (2.55) nm; CD (c 0.55, MeOH) λ(Δε): 199 (+7.27),
214 (+8.45), 247 (+0.42), 265 (+4.47), 282 (+3.28), 300
(+5.71) nm. 1H NMR (acetone-d6, 400 MHz) δH: 7.25
(2H, d, J = 7.4 Hz, H-2c/6c), 7.23 (2H, d, J = 7.4 Hz,
H-2a/6a), 7.05 (2H, d, J = 8.5 Hz, H-2b/6b), 6.93 (1H,
d, J = 1.6 Hz, H-14b), 6.80 (2H, d, J = 7.4 Hz, H-3a/5a),
6.79 (1H, J = 1.6 Hz, H-12b), 6.74 (2H, d, J = 7.4 Hz,
H-3c/5c), 6.69 (2H, d, J = 8.5 Hz, H-3b/5b), 6.54 (1H,
br d, J = 1.6 Hz, H-14a), 6.49 (1H, d, J = 2.0 Hz, H-
12c), 6.31 (1H, d, J = 2.0 Hz, H-14c), 6.24 (1H, d, J =
1.6 Hz, H-12a), 5.91 (2H, br s, H-7a/7b), 4.85 (1H, br s,
H-8b), 4.33 (1H, br s, H-8a); 13C NMR (acetone-d6, 100
MHz) δC: 163.6 (C-11c), 160.6 (C-13c), 160.0 (C-11a),
159.1 (C-13a), 158.3 (C-4c), 158.11 (C-4b), 158.05 (C-
4a), 157.1 (C-13b), 155.3 (C-11b), 149.6 (C-7c), 141.1
(C-9a), 139.8 (C-9b), 135.4 (C-9c), 133.6 (C-1a), 132.8
(C-1b), 128.4 (C-2b/6b), 128.3 (C-2c/6c), 127.4 (C-2a/
6a), 122.94 (C-10a), 122.87 (C-1c), 120.7 (C-10b), 119.2
(C-10c), 116.2 (C-3c/5c), 116.0 (C-3a/5a), 115.8 (C-3b/
5b), 114.6 (C-8c), 109.7 (C-14c), 108.71 (C-14a), 108.69
· 104 · 药学学报 Acta Pharmaceutica Sinica 2016, 51 (1): 100−104

(C-14b),98.4 (C-12c), 97.6 (C-12a), 96.4 (C-12b), 95.2
(C-7a), 88.0 (C-7b), 54.1 (C-8a), 46.0 (C-8b). The
key HMBC and NOESY correlations were showed in
Figure 3. (−)-ESI-MS m/z 675.25 [M−H]−.
Z-Miyabenol C (6): yellow amorphous powder;
C42H32O9; [α] 20D +82.22 (c 0.45, MeOH); UV (MeOH)
λmax (logε): 208 (3.23), 286 (2.71) nm; CD (c 0.67, MeOH)
λ (Δε): 220 (+1.81), 234 (−0.44), 251 (+3.53), 273 (−0.29),
286 (+0.30), 312 (−0.56), 342 (+0.15) nm. 1H NMR
(DMSO-d6, 600 MHz) δH: 7.00 (2H, d, J = 8.6 Hz,
H-2a/6a), 6.72 (2H, d, J = 8.6 Hz, H-3a/5a), 6.69 (2H, d,
J = 8.6 Hz, H-2c/6c), 6.47 (2H, d, J = 8.6 Hz, H-3c/5c),
6.46 (2H, d, J = 8.6 Hz, H-3b/5b), 6.31 (2H, d, J = 8.6
Hz, H-2b/6b), 6.28 (1H, d, J = 1.9 Hz, H-12c), 6.16 (1H,
d, J = 2.1 Hz, H-12b), 6.10 (1H, t, J = 2.1 Hz, H-12a),
6.06 (1H, d, J = 2.1 Hz, H-14b), 5.99 (1H, d, J = 1.9
Hz, H-14c), 5.88 (2H, d, J = 2.1 Hz, H-10a/14a), 5.74
(2H, d, J = 2.0 Hz, H-7c, 8c), 5.31 (1H, J = 1.9 Hz,
H-7b), 5.23 (1H, d, J = 3.2 Hz, H-7a), 4.22 (1H, d, J =
3.2 Hz, H-8a), 3.78 (1H, d, J = 1.9 Hz, H-8b); 13C NMR
(DMSO-d6, 150 MHz) δC: 160.7 (C-11b), 160.5 (C-11c),
159.8 (C-13b), 159.6 (C-11a/13a), 158.6 (C-13c), 157.7
(C-4a), 157.2 (C-4c), 156.8 (C-4b), 147.0 (C-9a), 142.4
(C-9b), 135.9 (C-9c), 133.2 (C-1a), 132.2 (C-1b), 130.8
(C-7c), 130.2 (C-2c/6c), 127.2 (C-1c), 127.0 (C-2a/6a),
125.9 (C-2b/6b), 124.7 (C-8c), 120.8 (C-10c), 118.1
(C-10b), 115.6 (C-3a/5a), 115.3 (C-3b/5b/3c/5c), 107.6
(C-14c), 106.1 (C-14b), 105.7 (C-10a/14a), 101.5 (C-12a),
96.5 (C-12c), 95.6 (C-12b), 92.7 (C-7a), 90.9 (C-7b),
55.8 (C-8a), 51.6 (C-8b). The key HMBC and ROESY
correlations were shown in Figure 4. (−)-ESI-MS m/z
679.15 [M−H]−.
References
[1] Chinese Pharmacopoeia Commission. Pharmacopoeia of the
People’s Republic of China: Vol 1 (中华人民共和国药典: 第
一卷 ) [S]. Beijing: China Medical Science Press, 2010:
79−80.
[2] Wu MH, Guo P, Tsui SW, et al. An ethnobotanical survey
of medicinal spices used in Chinese hotpot [J]. Food Res Int,
2012, 48: 226−232.
[3] Gellerstedt G, Lundquist K, Wallis AFA, et al. Revised
structures for neolignans from Arum italicum [J]. Phyto-
chemistry, 1995, 40: 263−265.
[4] Takeshi D, Takako I, Shizuka K, et al. The constituents of
Eucommia ulmoides OLIV. VI. Isolation of a new sesquilignan
and neolignan glycosides [J]. Chem Pharm Bull, 1987, 35:
1803−1807.
[5] Ma Y, Han GQ, Wang YY. PAF antagonistic benzofuran
neolignans from Piper kadsura [J]. Acta Pharm Sin (药学学
报), 1993, 28: 370−373.
[6] Na Z, Feng YL, Xu YK. Chemical constituents from aerial
parts of Eupatorium odoratum [J]. Chin Tradit Herb Drugs
(中草药), 2012, 43: 1896−1900.
[7] Kim DK, Lim JP, Kim JW, et al. Antitumor and antiinflammatory
constituents from Celtis sinensis [J]. Arch Pharm Res, 2005,
28: 39−43.
[8] Luo HF, Zhang LP, Hu CQ. Five novel oligostilbenes from
the roots of Caragana sinica [J]. Tetrahedron, 2001, 57:
4849−4854.
[9] Mattivi F, Vrhovsek U, Malacarne G, et al. Profiling of
resveratrol oligomers, important stress metabolites, accumulating
in the leaves of hybrid Vitis vinifera (Merzling × Teroldego)
genotypes infected with Plasmopara viticola [J]. J Agric Food
Chem, 2011, 59: 5364−5375.
[10] Wang XS, Che QM, Li YM, et al. A study on chemical
constituents in seeds of Crataegus pinnatifida Bge. var. major
N. E. Br. [J]. China J Chin Mater Med (中国中药杂志),
1999, 24: 739−740.
[11] Lin Y, Bao YY, Zhang YL, et al. Study on chemical
constituents of Taraxacum falcilobum Kitag [J]. Chin Tradit
Herb Drugs (中草药), 2000, 31: 10−11.
[12] Chen Y, Zhang GG, Mao DS, et al. Chemical constituents of
Scutellaria barbata D.Don (I) [J]. Chin J Med Chem (中国
药物化学杂志), 2008, 18: 48−50.