Four new phenylethanoid glycosides from Ternstroemia gymnanthera and their analgesic activities

Article abstract of DOI:10.1016/j.phytol.2019.09.009

Four new phenylethanoid glycosides, terngymnosides A–D (1–4), together with sixteen known compounds (5–20) were isolated from the aerial parts of Ternstroemia gymnanthera. Structures of the isolated compounds were determined by the comprehensive analyses of the physicochemical properties and spectral data (1D NMR, 2D NMR, and MS). Compounds 1–3, 8 and 10 showed significant analgesic effect with the inhibition rates ranging from 56.2% to 65.3% at a dose of 50 mg/kg, that was superior to the standard drug aspirin and was equivalent to indomethacin.

Full text of DOI:10.1016/j.phytol.2019.09.009

PhytochemistryLetters34(2019)25–29
Contents lists available at ScienceDirect
Phytochemistry Letters
journal homepage: www.elsevier.com/locate/phytol
Four new phenylethanoid glycosides from Ternstroemia gymnanthera and
their analgesic activities
Hua-Xuan Lia,b,1, Chao-Jiang Xiaoa,b,1, Min Wangb, Shu-Jun Cuib, Hai-Feng Lia, Kai-Ling Wanga,
⁎
Xiang Donga,b, Bei Jianga,b,
a Institute of Materia Medica, Dali University, Dali 671000, PR China
b College of Pharmacy and Chemistry, Dali University, Dali 671000, PR China
A R T I C L E I N F O
A B S T R A C T
Keywords:
Four new phenylethanoid glycosides, terngymnosides A–D (1–4), together with sixteen known compounds
(5–20) were isolated from the aerial parts of Ternstroemia gymnanthera. Structures of the isolated compounds
were determined by the comprehensive analyses of the physicochemical properties and spectral data (1D NMR,
2D NMR, and MS). Compounds 1–3, 8 and 10 showed significant analgesic effect with the inhibition rates
ranging from 56.2% to 65.3% at a dose of 50 mg/kg, that was superior to the standard drug aspirin and was
equivalent to indomethacin.
Ternstroemia gymnanthera
Theaceae
Phenylethanoid glycosides
Analgesic
1. Introduction
China of Chinese Academy of Science, 1998). In order to further un-
derstand the related analgesic active ingredients, guided by acetic acid
The genus Ternstroemia (Theaceae) comprises 90 species and is
mainly distributed in Central America, South America, Africa, and Asia
(Editorial Committee of the Flora of China of Chinese Academy of
Science, 1998). In China, a total of 14 species of Ternstroemia mainly
distributed in the south area of the Yangtze River are recorded, of
which two species, Ternstroemia gymnanthera (Wight et Arn.) Beddome
and T. luteoflora L. K. Ling, are used medicinally according to the China
Herbal (Editorial Board of China Herbal and State Administration of
Traditional Chinese Medicine, 1999; Editorial Committee of the Flora of
China of Chinese Academy of Science, 1998). Studies on Ternstroemia
plants are still limited, and during the past two decades, only a few
studies about the phytochemical content of Ternstroemia plants con-
cerning the isolation of several pentacyclic triterpenoids and their
glycosides (Ikuta et al., 2003; Shin et al., 2003), flavonoid glycosides
(Jo et al., 2006), and steroids (Ikuta et al., 2003) have been published.
In recent years, a systematic project to screen the analgesic plants from
Western Yunnan has been conducted in our research group. As part of
this work, Ternstroemia gymnanthera was found to possess definite an-
algesic activity. According to literatures, T. gymnanthera was a flow-
ering shrub or small tree (1.5 to 10 m) and could be used as folk
medicine by locals to treat carbuncles, sores, boils, and mastitis
(Editorial Board of China Herbal and State Administration of
Traditional Chinese Medicine, 1999; Editorial Committee of the Flora of
induced writhing model in mice, the active ingredients in 95% ethanol
extract from the aerial part of T. gymnanthera were tracked and studied.
As a result, four new phenylethanoid glycosides terngymnosides A–D
(1–4), along with 16 known compounds (5–20), were isolated and
structurally identified. The analgesic activities of compounds 1–10
were evaluated as well, and some compounds revealed significant po-
tential in analgesic activity.
2. Results and discussion
The 95% ethanol extract from the aerial part of T. gymnanthera was
partitioned with petroleum ether and ethyl acetate, successively. The
petroleum ether and EtOAc soluble portions were subjected to column
chromatography and preparative HPLC to yielded four new pheny-
lethanoid glycosides terngymnosides A–D (1–4, Fig. 1), along with 16
known compounds (5–20, Fig. S1). In addition, compounds 1–3, 8 and
10 showed significant analgesic activity.
Compound 1 was isolated as a brownish yellow solid, and assigned
the molecular formula C30H32O14 (fifteen unsaturation degrees) from
its HR-ESI-MS and 1H and 13C NMR spectra (including DEPT). The 1H
NMR spectrum of 1 (Table 1) showed one typical 4-oxo-2,5-cyclohex-
adien moiety at 7.06 (2H, d, J =10.2 Hz, H-2ꞌꞌ, 6ꞌꞌ) and 6.12 (2H, d, J
=10.2 Hz, H-3ꞌꞌ, 5ꞌꞌ), two ABX aromatic spin systems with protons at δH
⁎ Corresponding author at: Institute of Materia Medica, Dali University, Dali 671000, PR China.
E-mail address: jiangbei@dali.edu.cn (B. Jiang).
1 Both authors contributed equally to this work.
https://doi.org/10.1016/j.phytol.2019.09.009
Received 1 July 2019; Received in revised form 14 August 2019; Accepted 6 September 2019
1874-3900/©2019PhytochemicalSocietyofEurope.PublishedbyElsevierLtd.Allrightsreserved.

H.-X. Li, et al.
PhytochemistryLetters34(2019)25–29
Fig. 1. Structures of compounds 1–4.
Table 1
1H (400 MHz) and 13C (100 MHz) NMR spectroscopic data for compounds 1–4 in CD3OD.
No.
1
2
3
4
δH (mult., J in Hz)
δC
δH (mult., J in Hz)
δC
δH (mult., J in Hz)
δC
δH (mult., J in Hz)
δC
1
2
3
4
5
6
4.40 (d, 8.0)
101.9
75.0
75.5
71.4
74.8
64.6
4.40 (d, 8.1)
101.9
75.1
75.6
71.5
75.0
64.7
4.30 (d, 7.8)
104.4
75.0
71.5
77.8
75.1
65.0
4.34 (d, 8.0)
101.9
75.0
75.6
71.5
75.0
64.6
4.72 (dd, 9.2, 8.0)
3.56 (t, 9.2)
4.71 (dd, 9.5, 8.1)
3.54 (overlap)
3.19 (t-like, 8.4)
3.33 (overlap)
3.35 (overlap)
3.45 (m)
4.71 (dd, 9.2, 8.0)
3.54 (t, 9.2)
3.41 (t, 9.2)
3.39 (t, 9.1)
3.39 (t, 9.2)
3.48 (overlap)
4.44 (brd, 11.9)
4.21 (dd, 11.9, 5.2)
3.47 (overlap)
3.45 (overlap)
4.42 (dd, 11.9, 2.0)
4.23 (dd, 11.9, 5.9)
4.44 (dd, 12.0, 1.7)
4.20 (dd, 12.0, 5.5)
4.44 (dd, 11.9, 1.9)
4.18 (dd, 11.9, 5.8)
1'
2'
3'
4'
5'
6'
7'
8'
131.3
117.0
145.6
144.2
116.1
121.8
36.1
131.4
117.1
145.9
144.5
116.3
121.3
36.2
131.4
117.1
146.1
144.6
116.3
121.2
36.6
131.6
117.1
145.6
144.2
116.3
121.4
36.0
6.65 (d, 2.1)
6.64 (d, 2.0)
6.68 (d, 2.0)
6.65 (d, 2.1)
6.69 (d, 8.0)
6.68 (d, 8.0)
6.50 (dd, 8.0, 2.0)
2.56 (t, 7.3)
3.85 (m)
6.67 (d, 8.0)
6.70 (d, 8.1)
6.51 (dd, 8.0, 2.1)
2.55 (t, 7.0)
6.56 (dd, 8.0, 2.0)
2.79 (overlap)
6.51 (dd, 8.1, 2.1)
2.54 (m)
3.86 (q-like, 7.3)
3.50 (t, 7.8)
71.8
71.8
3.94 (dd, 16.4, 7.9)
3.68 (dd, 16.4, 8.3)
72.3
3.78 (dt, 9.4, 6.9)
3.45 (overlap)
71.7
3.49 (overlap)
1”
2”
3”
4”
5”
6”
7”
8”
1”'
2”'
3”'
4”'
5”'
6”'
7”'
8”'
67.9
68.0
68.0
126.7
117.3
145.9
145.1
116.1
121.7
41.1
7.06 (d, 10.2)
6.12 (d, 10.2)
152.6
128.1
187.4
128.1
152.6
45.5
7.07 (d, 10.4)
6.12 (d, 10.4)
152.7
128.2
187.5
128.2
152.7
45.6
7.06 (d, 10.1)
6.11 (d, 10.1)
152.7
128.3
187.4
128.3
152.7
45.8
6.74 (d, 2.1)
6.12 (d, 10.2)
7.06 (d, 10.2)
2.75 (s)
6.12 (d, 10.4)
7.07 (d, 10.4)
2.76 (s)
6.11 (d, 10.1)
7.06 (d, 10.1)
2.76 (s)
6.70 (d, 8.6)
6.57 (overlap)
3.48 (s)
170.2
126.8
117.5
145.9
145.1
116.2
121.3
41.3
170.2
126.3
131.4
116.1
157.2
116.1
131.4
41.1
170.3
173.9
126.8
117.5
145.9
145.1
116.2
121.8
41.2
6.77 (d, 2.0 Hz)
7.08 (d, 8.6)
6.74 (d, 8.6)
6.76 (d, 2.1)
6.73 (d, 8.1)
6.60 (dd, 8.1, 2.0)
3.46 (s)
6.74 (d, 8.6)
7.08 (d, 8.6)
3.51 (s)
6.72 (d, 8.5)
6.58 (overlap)
3.43 (s)
173.0
173.1
173.1
6.69 (1H, d, J =8.0 Hz, H-5ꞌ), 6.65 (1H, d, J =2.1 Hz, H-2ꞌ) and 6.51
(1H, dd, J = 8.0, 2.1 Hz, H-6ꞌ), and δH 6.77 (1H, d, J =2.0 Hz, H-2ꞌꞌꞌ),
6.73 (1H, d, J =8.1 Hz, H-5ꞌꞌꞌ) and 6.60 (1H, dd, J = 8.1, 2.0 Hz, H-
6ꞌꞌꞌ). In addition, nine protons from oxygenated methines, oxygenated
methylenes or hydroxyls at the range between δH 3.41 and 4.72, and six
proton resonances at δH 2.55 (2H, t, J =7.0 Hz), 2.75 (2H, s) and 3.46
(2H, s) from three methylenes were also observed. Inspection of the 13
C
NMR spectrum of 1 with the aid of the DEPT-90 and -135 spectra re-
vealed the existence of 30 carbon resonances (Table 1), including five
sp3 methylenes (including two oxy-methylenes at δC 64.6 and 71.8),
five sp3 oxy-methines, ten sp2 methines, one sp3 quaternary carbon (δC
67.9), and nine sp2 quaternary carbons (including three carbonyls at δC
26

H.-X. Li, et al.
PhytochemistryLetters34(2019)25–29
Fig. 2. Key HMBC, 1H-1H COSY and ROESY correlations of compounds 1 and 2.
170.2, 173.0 and 187.4). Comparison of the spectral data with those of
ternstroside A (Jo et al., 2006) and quinolacetic acid (Brownlee et al.,
2010) previously reported, 1 was assumed to be an adduct of the two
compounds.
67.9, 128.1 (double), 152.6 (double), 170.2 and 187.4) in 1 shifted to
these of a typical 3,4-dihydroxyphenylacetic acid moiety at δC 41.1,
116.1, 117.3, 121.7, 126.7, 145.1, 145.9 and 173.9 in 4. Further ana-
lyses of 2D NMR spectral data (1H-1H COSY, HMBC and ROESY, Fig. 3),
the specific rotation data and analysis of acid hydrolysis of compound 4
allowed the spatial structure to be established. Therefore, 4 was char-
acterized as shown in Fig. 1, and named terngymnoside D.
Analysis of 1H–1H COSY spectrum (Fig. 2) confirmed that 1 pos-
sessed an aldohexose, judging from the correlations of H-1 (δH 4.40, d, J
=8.0 Hz)/H-2 (δH 4.72, dd, J = 9.2, 8.0 Hz)/H-3 (δH 3.56, t,
J
=9.2 Hz)/H-4 (δH 3.41, t, J =9.2 Hz)/H-5 (δH 3.48, overlap)/H2-6 (δH
4.21, dd, J = 11.9, 5.2 Hz; δH 4.44, brd, J =11.9 Hz). The coupling
constants of H-2 (dd, J = 9.2, 8.0 Hz), H-3 (t, J =9.2 Hz) and H-4 (t, J
=9.2 Hz) suggested the aldohexose in 1 should be glucose. Then the β-
configuration of the glucose was determined by coupling constant of H-
1 (d, J =8.0 Hz) (Pretsch et al., 2009). The H-1 and H-2 showed HMBC
correlations (Fig. 2) to C-8' (δC 71.8) and C-8”' (δC 173.0), respectively,
which confirmed the ternstroside A moiety in 1. Similarly, the quino-
lacetic acid moiety linked to C-6, owing to the HMBC correlation
(Fig. 2) between H2-6 and C-8” (δC 170.2). For identification of ste-
reochemistry of the sugar, acid hydrolysis of 1 was performed. The
stereochemistry of the glucose was determined as D-configuration by
comparing its specific rotation data with that of the authentic sugar
glucose. Therefore, compound 1 was determined as shown in Fig. 1, and
named terngymnoside A.
By comparison of their spectroscopic data and physicochemical
properties with those reported in the literature, the sixteen known
compounds were identified as 6′-O-coumaroyl-1′-O-[2-(3,4- dihydrox-
yphenyl)ethyl]-β-D-glucopyranoside (5) (Es-Safi et al., 2005), ternstro-
side A (6) (Jo et al., 2006), ternstroside D (7) (Jo et al., 2006), 3',4'-
dihydroxyphenethyl alcohol 1-O-β-D-glucopyranoside (8) (Yahara et al.,
1985), jacaranone (9) (Jo et al., 2005), α-tocopherol (10) (Ponrasu
et al., 2008), 1,6-caprolactam (11) (Wang et al., 2009), stearic acid (12)
(Xiao et al., 2013), oleic acid (13) (Zhao et al., 2017), linoleic acid (14)
(Marwah et al., 2007), oleanolic acid (15) (Acebey-Castellon et al.,
2011), ursolic acid (16) (Ibrahim et al., 2008), kajiichigoside F1 (17)
(Guang-Yi et al., 1989; Seto et al., 1984), β-sitosterol (18) (Cho et al.,
2012), daucosterol (19) (Cho et al., 2012) and spinasterol (20) (Kojima
et al., 1990), respectively.
Compounds 1–10 were tested for their in vivo analgesic activities by
acetic acid induced writhing assay. As shown in Table 2, phenyletha-
noid glycosides compounds 1–3 and 8, as well as the antioxidant α-
tocopherol (10) showed significant analgesic effect with the inhibition
rates ranging from 56.2% to 65.3% at a dose of 50 mg/kg, which was
superior to the standard drug aspirin (inhibition rate of 55.3% at
200 mg/kg) and was equivalent to indomethacin (inhibition rate of
58.2% at 50 mg/kg). The analgesic activities were reduced when the
substituent groups at glucopyranose (compound 8 vs 4–7). However,
the quinolacetic acid moiety is an important substituent group for
phenylethanoid glycosides to exert their activities, due to compounds
1–3 showed stronger activity than 4.
Compound 2 was isolated as a brownish yellow solid, and possessed
a molecular formula C30H32O13 (fifteen unsaturation degrees) ac-
cording to its HR-ESI-MS and NMR data. The 1H and 13C NMR spectral
data of 2 (Table 1) were very similar to those of 1, indicating that 2 was
also a phenylethanoid glycoside derivative. However, compound 2
significantly differed from 1 by the replacement of a quaternary carbon
(δC 145.9 for C-3”') in 1 by a sp2 methine in 2 (δC 116.1), implying that
2 should be a C-3”' deoxy derivative of 1. Based on the analyses of
1H-1H COSY, HMBC and ROESY spectra (Fig. 2), as well as the specific
rotation data of compound 2, compound 2 was finally assigned as
shown in Fig. 1, and named terngymnoside B.
Compound 3 was isolated as a brownish yellow solid, and was es-
tablished the molecular formula C22H26O11 (ten unsaturation degrees)
according to its HR-ESI-MS and NMR data. The 1H and 13C NMR
spectral data of 3 (Table 1) were very similar to those of 1, indicating
that 3 was an analogue of 1. However, compound 3 significantly dif-
fered from 1 by lack of the 3,4-dihydroxyphenylacetic acid signals in
NMR spectra, implying that the C-2 was unsubstituted in 3. According
to the analyses of 1H-1H COSY, HMBC and ROESY spectra (Fig. 3), as
well as the specific rotation data of compound 3, compound 3 was fi-
nally assigned as shown in Fig. 1, and named terngymnoside C.
Compound 4 was isolated as a brownish yellow solid, and possessed
a molecular formula C30H32O14 (fifteen unsaturation degrees) ac-
cording to its HR-ESI-MS and NMR data. The 1H and 13C NMR spectral
data of 4 (Table 1) were very similar to those of 1, and the major dif-
ference was the chemical shifts of the quinolacetic acid moiety (δC 45.5,
3. Experimental
3.1. General experimental procedures
NMR spectra were recorded on a Bruker Avance III-400 instrument
(Bruker, Karlsruhe, Germany) with TMS as an internal reference. HR-
ESI-MS data were obtained on a Dionex Ultimate 3000 LC System
(Thermo Fisher Scientific, Sunnyvale, USA) coupled in series to a
Bruker Compact quadrupole time-of-flight (QTOF) mass spectrometer
in ESI-mode (Bruker, Bremen, Germany). Optical rotations were de-
termined on a SGW-3 automatic polarimeter (Shanghai INESA Physico
optiacal instrument Co., Ltd, Shanghai, P. R. China). UV data were
obtained on a TU-1901 UV/Vis spectrophotometer (Beijing Purkinje
General Instrument Co. Ltd., Beijing, P. R. China). IR spectra were
27

H.-X. Li, et al.
PhytochemistryLetters34(2019)25–29
Fig. 3. Key HMBC, 1H-1H COSY and ROESY correlations of compounds 3 and 4.
(B1–B3). Subfraction B1 was subjected to repeated silica gel CC eluting
with PE-acetone (40:1, v/v) and CHCl3-MeOH (100:1, v/v) to yield
compounds 15 (12 mg) and 16 (5 mg). Subfraction B3 was purified by a
silica gel CC eluting with CHCl3-MeOH (50:1, v/v), a Sephadex LH-20
CC (CHCl3-MeOH, 1:1, v/v) and a preparative HPLC (MeOH-H2O, 4:6,
v/v) to yield compounds 2 (70 mg), 11 (7 mg), 17 (30 mg) and 19
(5 mg). Fraction C (200 g) was purified by a silica gel CC (CHCl3-
acetone, 20:1 to 0:1, v/v) and repeated preparative HPLC (MeOH-H2O,
25%–45%, v/v) to give compounds 1 (140 mg), 3 (19 mg), 4 (16 g), 5
(20 mg), 6 (24 mg) and 7 (40 mg). Fraction D (100 g) was purified by a
silica gel CC (CHCl3-acetone, 5:1 to 1:1, v/v) and repeated preparative
HPLC (MeOH-H2O, 25%–45%, v/v) to give compounds 8 (410 mg) and
9 (18 mg).
Table 2
In vivo analgesic activities of compounds 1–10 (n = 5).
Sample
Dose (mg/kg)
Number of writhes (
x
s)
Inhibition (%)
Control
–
39.5
15.7
15.3
13.7
26.0
21.0
20.0
20.3
17.3
20.0
17.7
16.5
17.7
1.3
–
1
50
50
50
50
50
50
50
50
50
50
50
200
3.5*
2.1*
1.2*
3.6*
1.8*
2.6*
1.0*
1.2*
3.2*
1.5*
6.6*
3.2*
60.2
61.2
65.3
34.1
46.8
49.3
48.6
56.2
49.3
55.3
58.2
55.3
2
3
4
5
6
7
8
9
10
Indomethacin
Aspirin
The PE soluble portion (180 g) was divided into eight fractions
(I–VIII) by silica gel CC eluting with gradient PE-EtOAc (1:0 to 0:1, v/
v). Fraction IV (24 g) was subjected to repeated silica gel CC with PE-
EtOAc (80:1 to 0:1, v/v) to afford compound 10 (310 mg). Fraction VI
(6.5 g) was chromatographed on a MCI gel column (MeOH-H2O, 1:1 to
1:0, v/v) and a Sephadex LH-20 column (CHCl3-MeOH, 1:1, v/v), suc-
cessively, then was further purified by a silica gel column with PE-
EtOAc (20:1, v/v) to give compounds 12 (16 mg), 13 (18 mg), 14
(19 mg) and 18 (60 mg).
Control: 0.5% CMC-Na. Compared with the control group, *P < 0.05.
recorded by a Nicolet 380 FT-IR spectrophotometer (Thermo Scientific,
Madison, WI, USA) with KBr pellets. Preparative HPLC analysis was
performed on a Shimadzu LC-20AP series instrument (Shimadzu, Kyoto,
Japan). Silica gel (Qingdao Marine Chemical Ltd., Qingdao, P. R.
China), Sephadex LH-20 (Amersham Biosciences, Uppsala, Sweden) and
MCI gel (75–150 μm; Mitsubishi Chemical Corp., Tokyo, Japan) were
used for open column chromatography. Silica gel GF254 plates (Qingdao
Marine Chemical Ltd.) were used for thin layer chromatography (TLC)
detection.
3.3.1. Terngymnoside A (1)
Brownish yellow solid; [ ]D22 -29.3 (c = 0.1, MeOH); UV (MeOH)
λmax (log ε): 223 (4.27), 291 (3.82) nm; IR (KBr) νmax: 3375, 2953,
1730, 1617, 1522, 1445, 1282, 1179, 1076, 862, 802, 628 cm−1; HR-
ESI-MS m/z: 615.1725 [M−H]− (calcd for C30H31O14, 615.1719). 1H
and 13C NMR spectroscopic data see Table 1.
3.2. Plant material
The aerial parts of Ternstroemia gymnanthera (Wight et Arn.)
Beddome were collected in Yangbi County, Yunnan Province, P. R.
China, in July 2017 and identified by Dr. De-Quan Zhang at the College
of pharmacy and chemistry, Dali University, P. R. China. A voucher
specimen (NO. 2015 1008-6) has been deposited at the Institute of
Materia Medica, Dali University.
3.3.2. Terngymnoside B (2)
Brownish yellow solid; [ ]D22 -27.6 (c = 0.1, MeOH); UV (MeOH)
λmax (log ε): 224 (4.33), 285 (3.75) nm; IR (KBr) νmax: 3374, 2932,
1730, 1609, 1519, 1445, 1365, 1273, 1166, 1077, 862, 810, 619 cm−1
;
HR-ESI-MS m/z: 599.1761 [M−H]− (calcd for C30H31O13, 599.1770).
1H and 13C NMR spectroscopic data see Table 1.
3.3. Extraction and isolation
3.3.3. Terngymnoside C (3)
Brownish yellow solid; [ ]D22 -26.0 (c = 0.1, MeOH); UV (MeOH)
λmax (log ε): 223 (4.22), 291 (3.84) nm; IR (KBr) νmax: 3404, 2946,
2824, 1728, 1607, 1519, 1445, 1362, 1282, 1168, 1078, 776,
The air-dried aerial parts (14 kg) of T. gymnanthera were crushed
and soaked with 95% ethanol (60 L ×12 h × 8) at room temperature.
The ethanol extracts were evaporated to dryness under reduced pres-
sure by a rotary evaporator, and the residue (2.3 kg) was suspended in
water and partitioned with petroleum ether (PE) and ethyl acetate,
successively. The EtOAc soluble portion (726 g) was divided into seven
fractions (A–G) by silica gel column chromatography (CC) eluting with
gradient chloroform-methanol (1:0 to 0:1, v/v). Fraction A (16 g) was
chromatographed on a MCI gel column (MeOH-H2O, 1:1 to 1:0, v/v) to
yield compound 20 (40 mg). Fraction B (200 g) was subjected to a silica
gel CC (CHCl3-acetone, 9:1 to 0:1, v/v) to yield three subfractions
614 cm−1; HR-ESI-MS m/z: 465.1405 [M−H]− (calcd for C22H25O11
,
465.1402). 1H and 13C NMR spectroscopic data see Table 1.
3.3.4. Terngymnoside D (4)
Brownish yellow solid; [ ]D22 −60.0 (c = 0.1, MeOH); UV (MeOH)
λmax (log ε): 220 (4.38), 292 (3.99) nm; IR (KBr) νmax: 3369, 1725,
1607, 1524, 1446, 1363, 1286, 1194, 1113, 1079, 1011, 964, 798,
608 cm−1; HR-ESI-MS m/z: 615.1702 [M−H]− (calcd for C30H31O14
,
615.1719). 1H and 13C NMR spectroscopic data see Table 1.
28

H.-X. Li, et al.
PhytochemistryLetters34(2019)25–29
3.4. Acid hydrolysis for identification of sugars
online version, at doi:https://doi.org/10.1016/j.phytol.2019.09.009.
The compounds 1 (20 mg) and 4 (50 mg) was dissolved in 5 or
10 mL 10% hydrochloric acid aqueous solution and heated at 60 °C for
10 h, respectively. The monosaccharide was separated and purified on
silica gel column (chloroform-methanol, 3:1 to 0:1), and analyzed by
co-TLC with the authentic sugar glucose (Rf 0.6; chloroform-methanol-
water, 5:5:1). The stereochemistry of the monosaccharide was de-
termined by comparing its specific rotation data with that of the au-
thentic sugar glucose.
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Declaration of Competing Interest
The authors declare that there is no conflict of interest.
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Appendix A. Supplementary data
Supplementary material related to this article can be found, in the
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