全 文 ：Received 3 Feb. 2004 Accepted 29 Sept. 2004
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Acta Botanica Sinica
植 物 学 报 2004, 46 (12): 1454-1457
A New Monoterpene Glycoside from Hemiphragma heterophyllum
YANG Ming-Fu1, 2, LI Yu-Yuan1, LI Bo-Gang1, ZHANG Guo-Lin1*
(1. Chengdu Institute of Biology, The Chinese Academy of Sciences, Chengdu 610041, China;
2. Chengdu Institute of Organic Chemistry, The Chinese Academy of Sciences, Chengdu 610041, China)
Abstract: A new monoterpene glycoside, hemiphroside C (1), along with seven known compounds were
isolated from the whole plants of Hemiphragma heterophyllum Wall. By spectral evidence, the structure of
the new compound was elucidated as (4S)-a-terpineol 8-O-b-D-xylopyranosyl-(1→6)-b-D-glucopyranoside.
The known compounds were identified to be globularin (2), (2S, 3S, 4R, 9E)-1, 3, 4-trihydroxy-2-[(2R)-
hydroxytetracosanoylamino]-9-octadecene (3), b-amyrin (4), oleanolic acid (5), cinnamic acid (6), b-sitosterol
(7) and daucosterol (8). All compounds except compound 2 were firstly isolated from H. heterophyllum.
Key words: Hemiphragma heterophyllum; a-terpineol; hemiphroside C; ceramide
Hemiphragma heterophyllum, a unique species of the
genus Hemiphragma (Scrophulariaceae), is used in folk
medicine for the treatment of cholecystitis, rheumatism,
abnormal menstruation, toothache in China (Jiangsu Insti-
tute of Botany, 1990). Phytochemical study on the whole
plants of H. heterophyllum led to the isolation of four
phenylpropanoid glycosides and three iridoid glycosides
from the n-BuOH soluble fraction of methanolic extract (Ma
et al., 1995). In this study, eight compounds, including one
new monoterpene glycoside were isolated from the ethanolic
extract of the whole plants of H. heterophyllum. Based on
the spectral analysis, the new compound was elucidated as
(4S)-a-terpineol 8-O-b-D-xylopyranosyl-(1®6)-b-D-
glucopyranoside, namely hemiphroside C (1). The known
ones were identified as globularin (2) (Ma et al., 1995), (2S,
3 S , 4 R , 9 E ) - 1 , 3 , 4 - t r i h y d r o x y - 2 - [ ( 2 R ) -
hydroxytetracosanoylamino]-9-octadecene (3) (Su et al.,
2002), b-amyrin (4) (Seo et al., 1981), oleanolic acid (5)
(Maillard et al., 1992), cinnamic acid (6), b-sitosterol (7)
and daucosterol (8).
1 Results and Discussion
Compound 1 was obtained as white amorphous powder.
ESI mass spectra exhibited ion peak at m/z 447 [M – H]- and
471 [M + Na]+. Its molecular formula C21H36O10 was con-
sistent with the ion peak at m/z 471.219 7 [M+Na]+ (cacld.
for C21H36O10Na 471.220 6) in HRESIMS spectrum. IR ab-
sorption at 3 300 cm-1 and 1 600 cm-1 indicated the pres-
ence of hydroxyl and double bond, respectively. 1H-NMR
signals at the range of 3.0 to 4.6 and eleven 13C-NMR sig-
nals at the range of 65-105 suggested the existence of
disaccharides. The acid hydrolysis of compound 1
provided D-glucose and D-xylose.
Besides signals for sugars, ten additional signals, in-
cluding three methyls, three methylenes, two methines, and
two quartery carbons were recognized in 13C-NMR
spectrum. 1H-NMR signals displayed the presence of two
methyls at d 1.24 and 1.19, one vinyl methyl at d 1.63, and
one olefinic proton at d 5.37. By comparing the 1H- and 13C-
NMR data with those of (4R)- and (4S)-a-terpineol glyco-
sides (Voirin et al., 1990; Nagatani et al., 2002), compound
1 could be determined as a (4S)-a-terpineol disaccharides
glycoside.
In 1H-NMR spectrum, two anomeric protons resonated
at d 4.48 (1H, d, J = 7.7 Hz), 4.30 (1H, d, J = 7.4 Hz), indicating
b-configurated sugar moieties. The latter was assigned to
H-1" of b-D-xylopyranosyl based on the HMBC correla-
tion of this one with C-5" (d 65.6, t). So the former was
assigned to H-1 of b-D-glucopyranosyl. The HMBC corre-
lation of H-1" with C-6 (d 68.7, t) suggested 1→6 linkage
of b-D-xylopyranosyl and b-D-glucopyranosyl. b-D-
xylopyranosyl-(1®6)-b-D-glucopyranosyl located to C-8
on the basis of the HMBC correlation between H-1 and C-
8 (d 79.9, s). Therefore, Compound 1 was elucidated as (4S)-
a-terpineol 8-O-b -D-xylopyranosyl-(1®6)-b -D-
glucopyranoside.
2 Experimental
2.1 General experimental procedures
Optical rotation was measured using a Perkin-Elmer 241
automatic polarimeter in methanol. Melting points were
determined on an XRC-1 micro-melting point apparatus and
uncorrected. IR (KBr disc) and UV (CH3OH as solvent)
spectra were recorded on Spectrum One FT-IR spectrometer
YANG Ming-Fu et al.: A New Monoterpene Glycoside from Hemiphragma heterophyllum 1455
and Lambda 35 spectrometer, respectively. MS spectra were
obtained on Finnigan LCQDECA ion trap mass spectrometer
(ESI) and API Q-STAR PULSAR i mass spectrometer
(HRESI). NMR spectra were recorded on Advance 600
spectrometer, tetramethylsilane (TMS) as internal reference.
Chemical shifts were given in ppm (d ) and coupling con-
stants were reported in Hz. Silica gel H (160-200, 200-300,
Qingdao Ocean Chemical Factory, Qingdao, China), silica
gel 60 (Merck, 230-400 mesh), and MCI gel (Mitsubishi,
75-150 m) were used for column chromatography. Organic
solvents were distilled prior to use.
2.2 Plant materials
The whole plants of Hemiphragma heterophyllum Wall.
were collected in December 2001 from Luding County,
Sichuan Province of China, and identified by Prof. ZHAO
Zuo-Cheng in Chengdu Institute of Biology, The Chinese
Academy of Sciences (CAS). A voucher specimen (No. HH-
1) is deposited at the Herbarium of Chengdu Institute of
Biology, CAS.
2.3 Extract and isolation
Air-dried and powdered whole plants (2.9 kg) was per-
colated three times with 90% EtOH (25 L× 7 d each) at
room temperature to give a dark residue (98 g). The residue
was suspended in water and partitioned with CHCl3, EtOAc,
respectively.
The EtOAc fraction (6.8 g) was subjected to column
chromatography (8× 50 cm) over silica gel (800 g) with
CHCl3/ CH3OH gradients (10:1, 3 L; 5:1, 2L; 1:1, 3 L) to yield
fractions f1-f3. Fraction f2 (860 mg) was separated on RP-
18 column chromatography eluted by CH3OH:H2O (30:70).
The eluate of 550-700 mL and 1 200-1 450 mL were con-
centrated to give 36 mg and 126 mg mixture, respectively.
Separated on silica gel 60, the former gave compound 1 (16
mg) and the latter yielded globularin (2) (48 mg). Fraction f1
(1.53 g) and f3 (2.60 g) were rechromatographied on silica
gel column to give (2S, 3S, 4R, 9E)-1, 3, 4-trihydroxy-2-[(2R)-
hydroxytetra cosanoylamino]-9-octadecene (3) (15 mg) and
daucosterol (8) (860 mg), respectively. The CHCl3 fraction
(35 g) was subjected to MCI gel column chromatography
using MeOH:H2O (80:20, 5 L) and 100% CH3OH (3.5 L) as
eluent. From the eluate of 80% CH3OH, b-amyrin (4) (12
mg)，oleanolic acid (5) (16 mg), cinnamic acid (6) (23 mg),
and b -sitosterol (7) (1 .36 g) were isolated by
rechromatographying over silica gel column.
2.4 Identification
Hemiphroside C (1) White amorphous powder. Mp
189.2-190.0 °C. [a]20D -48.7 ° ( c 0.34, CH3OH). UV lMeOH
(log e) nm: 211 (4.2); IR nKBr cm-1: 3 428, 2 923, 1 633, 1 367,
1 163, 1 079, 1 039; 1H-NMR (CD3OD, 600 MHz) d: 5.37 (1 H,
br.s, H-2), 2.06 (1 H, m, H-3), 1.80 (1 H, m, H-3), 1.71 (1 H, m,
H-4), 2.04 (2 H, m, H-5), 2.04 (1 H, m, H-6), 1.93 (1 H, m, H-6),
1.63 (3 H, s, H-7), 1.19 (3 H, s, H-9), 1.24 (3 H, s, H-10), 4.48 (1
H, d, 7.7 Hz, H-1), 3.18 (1 H, m, H-2), 3.36 (1 H, m, H-3), 3.37
(1 H, m, H-4), 3.38 (1 H, m, H-5), 4.00 (1 H, m, H-6), 3.76 (1 H,
m, H-6), 4.30 (1 H, d, 7.4 Hz, H-1"), 3.22 (1 H, m, H-2"), 3.31
(1 H, m, H-3"), 3.48 (1 H, m, H-4"), 3.85 (1 H, m, H-5"), 3.20 (1
H, m, H-5"); 13C-NMR (CD3OD, 150 MHz) d : 133.7 (s, C-1),
120.7 (d, C-2), 26.9 (t, C-3), 43.9 (d, C-4), 23.9 (t, C-5), 30.9 (t,
C-6), 22.4 (q, C-7), 79.9 (s, C-8), 21.8 (q, C-9), 24.0 (q, C-10),
97.4 (d, C-1), 74.1 (d, C-2), 77.0 (d, C-3), 70.3 (d, C-4), 75.2
(d, C-5), 68.7 (t, C-6), 104.3 (d, C-1"), 73.6 (d, C-2"), 76.4 (d,
C-3"), 70.0 (d, C-4"), 65.6 (d, C-5"); (+)ESIMS m/z 471
[M+Na]+, (-)ESIMS m/z 447 [M-H]-; HRESIMS m/z
471.219 7 [M+Na]+ (calcd. for C21H36O10Na 471.220 76).
Globularin (2) White amorphous powder. Mp 116.3-
118.2 °C. [a]20D -68.2° (c 0.20, MeOH). ESIMS m/z 493
[M+1]+, 515 [M+Na]+ (positive mode); IR nKBr cm-1: 3 427,
2 921, 1 700, 1 636, 1 451, 1 332, 1 315, 1 184, 1 076, 1 042,
1 016, 770, 686; 1H-NMR (Me2CO-d6, 500 MHz) d: 4.98 (d,
8.2, H-1), 5.10 (m, H-4), 2.31 (m, H-5), 2.59 (dd, J = 7.8 Hz,
8.0 Hz, H-9), 5.08 (d, J = 8.2 Hz, H-1), 7.43-7.71 (m, H-2, 3,
4, 5, 6), 6.60 (1H, d, J = 16.0 Hz, H-8), 7.74 (1H, d, J = 16.0
Hz, H-7); 13C-NMR (Me2CO-d6, 125 MHz) d: 95.1 (d, C-1),
140.9 (d, C-3), 103.4 (d, C-4), 38.3 (d, C-5), 78.6 (d, C-6), 62.2
Fig.1. Major HMBC correlation for hemiphroside C (1).
max
max
max
Acta Botanica Sinica 植物学报 Vol.46 No.12 20041456
(d, C-7), 62.5 (s, C-8), 42.8 (d, C-9), 63.3 (t, C-10), 134.9 (s, C-
1), 129.4 (d, C-2, 6), 128.7 (d, C-3, 5), 130.8 (d, C-4), 145.4
(s, C - 7), 118.3 (s, C -8), 166.9 (s, C = O), 99.9 (d, C-1),
74.1 (d, C-2), 77.3 (d, C-3), 71.6 (d, C-4), 77.1 (d, C-5),
61.8 (t, C-6).
(2S, 3S, 4R, 9E)-1, 3, 4-Trihydroxy-2-[(2R)-hydroxytet-
racosanoylamino]-9-octadecene (3) White amorphous
powder. Mp 141.3-143.3 oC. [a]20D +13.6o (c 0.36, MeOH).
ESIMS m/z 682 [M+H]+ (positive mode); IR nKBr cm-1:
3 336, 2 919, 2 850, 1 622, 1 544, 1 467, 1 066, 722; 1H-NMR
(400 MHz, pyridine-d5) d : 8.57 (d, 8.7 Hz, -NH), 5.48 (m, H-
9, 10), 5.10 (dd, 4.8 Hz, 4.5 Hz, H-2), 4.60 (dd, 7.2 Hz, 4.0 Hz,
H-2), 4.47 (m, H-1a), 4.41 (m, H-1b), 4.38 (m, H-3), 4.26 (m, H-
4), 2.23-2.44 (m), 1.96 (m), 1.28 (br.s), 0.83 (t, 6.9 Hz, H-18,
24); 13C-NMR (100 MHz, pyridine-d5) d : 175.3 (s, C-1),
130.8 (d, C-9 or 10), 130.6 (d, C-10 or 9), 76.8 (d, C-3), 72.3 (d,
C-4), 72.9 (CH, C-2), , 61.9 (t, C-1), 52.9 (d, C-2), 35.6 (t, C-3),
34.1 (t, C-5), 33.8 (t, C-8), 33.2 (t, C-11), 32.9 (t, C-16), 32.1 (t,
C-22), 30.3-29.5 (t), 26.7 (t, C-4), 25.8 (t, C-6), 22.9 (t, C-17,
23), 14.2 (q, C-18, 24).
b-Amyrin (4) White amorphous powder. Mp 185.0-
187.0 oC. Its UV, IR and NMR data were identical (resemble)
to those reported. This compound was identified by com-
paring its TLC behavior with authentic sample.
Oleanolic acid (5) White amorphous powder. Mp
308.2-309.0 oC. The identification was succeeded by the
identical TLC behavior and the same NMR data of com-
pound 5 with those of authentic sample.
Cinnamic acid (6) Colorless needles (CHCl3).
Mp136.0-138.0 oC. This compound was characterized by
the closely resemble NMR data and TLC behavior with
those of cinnamic acid.
b-Sitosterol (7) Colorless needles (CHCl3). Mp 136.0-
138.2 oC. By comparing its NMR data and TLC behavior of
compound 7 with those of b-sitosterol, this compound was
determined to be b-sitosterol.
Daucosterol (8) White amorphous powder. Mp
295.3-297.0 oC. The equal NMR data and TLC behavior of
compound 8 and daucosterol suggested this compound
could be daucosterol.
Acknowledgements: We are grateful to Prof. YIN Kai-Pu
(Chengdu Institute of Biology, The Chinese Academy of
Sciences (CAS)) for collecting plants and Prof. ZHAO Zuo-
Cheng (Chengdu Institute of Biology, CAS) for identifying
the sample.
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(Managing editor: WANG Wei)
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