New furostanol glycosides from Polygonatum multiflorum (L.) All

Article abstract of DOI:10.1080/14786419.2018.1431628

The phytochemical investigation of the whole plant of Polygonatum multiflorum resulted in the isolation of two new steroidal glycosides, polmultoside A (4) and polmultoside B (5), along with three known glycosides protobioside (1), protodeltonin (2) and huangjiangsu A (3). The structures of the isolated compounds have been elucidated by extensive 1D (¹H, ¹³C) and 2D (COSY, HSQC, HMBC) NMR spectral data analysis, as well as high-resolution mass determinations.

Full text of DOI:10.1080/14786419.2018.1431628

Natural Product Research
Formerly Natural Product Letters
ISSN: 1478-6419 (Print) 1478-6427 (Online) Journal homepage: http://www.tandfonline.com/loi/gnpl20
New furostanol glycosides from Polygonatum
multiflorum (L.) All.
Lili Gvazava, Vazha Nebieridze, Markus Ganzera & Alexander Skhirtladze
To cite this article: Lili Gvazava, Vazha Nebieridze, Markus Ganzera & Alexander Skhirtladze
(2018): New furostanol glycosides from Polygonatum multiflorum (L.) All., Natural Product
Research, DOI: 10.1080/14786419.2018.1431628
To link to this article: https://doi.org/10.1080/14786419.2018.1431628
View supplementary material
Published online: 31 Jan 2018.
Submit your article to this journal
View related articles
View Crossmark data
Full Terms & Conditions of access and use can be found at
http://www.tandfonline.com/action/journalInformation?journalCode=gnpl20

Natural Product research, 2018
ꢁꢁpꢂ://ꢃꢄi.ꢄꢅg/10.1080/14786419.2018.1431628
ꢀ
New furostanol glycosides from Polygonatum multiflorum (L.)
All.
a
a
b
a
Lili Gvazava , Vazha Nebieridze , Markus Ganzera and Alexander Skhirtladze
ꢆ
dꢇpꢆꢅꢁmꢇnꢁ ꢄf Pꢀyꢁꢄꢈꢀꢇmiꢂꢁꢅy, Iꢄvꢇꢉ Kꢊꢁꢆꢁꢇꢉꢆꢃzꢇ Inꢂꢁiꢁꢊꢁꢇ ꢄf Pꢀꢆꢅmꢆꢈꢄꢈꢀꢇmiꢂꢁꢅy, tbiꢉiꢂi sꢁꢆꢁꢇ Mꢇꢃiꢈꢆꢉ
b
univꢇꢅꢂiꢁy, tbiꢉiꢂi, Gꢇꢄꢅgiꢆ; Inꢂꢁiꢁꢊꢁꢇ ꢄf Pꢀꢆꢅmꢆꢈy, Pꢀꢆꢅmꢆꢈꢄgnꢄꢂy, univꢇꢅꢂiꢁy ꢄf Innꢂbꢅꢊꢈk, Innꢂbꢅꢊꢈk, aꢊꢂꢁꢅiꢆ
ABSTRACT
ARTICLE HISTORY
The phytochemical investigation of the whole plant of Polygonatum
multiflorum resulted in the isolation of two new steroidal glycosides,
polmultoside A (4) and polmultoside B (5), along with three known
glycosides protobioside (1), protodeltonin (2) and huangjiangsu A (3).
The structures of the isolated compounds have been elucidated by
extensive 1D ( H, C) and 2D (COSY, HSQC, HMBC) NMR spectral data
analysis, as well as high-resolution mass determinations.
rꢇꢈꢇivꢇꢃ 10 oꢈꢁꢄbꢇꢅ 2017
aꢈꢈꢇpꢁꢇꢃ 19 Jꢆnꢊꢆꢅy 2018
KEYWORDS
Pylygonatum multiflorum;
cꢄnvꢆꢉꢉꢆꢅiꢆꢈꢇꢆꢇ; ꢂꢁꢇꢅꢄiꢃꢆꢉ
gꢉyꢈꢄꢂiꢃꢇꢂ; fꢊꢅꢄꢂꢁꢆnꢄꢉꢂ;
ꢃiꢄꢂgꢇnin; pꢄꢉmꢊꢉꢁꢄꢂiꢃꢇ a-B
1
13
1
. Introduction
The genus Polygonatum (Convallariaceae) contains about 60 species, most of which occur
in the North Temperate Zone, with 5 species in Georgia (Gagnidze 2011). Species of
Polygonatum showed aphrodisiac (Kazmi et al. 2012), antimicrobial (Khan et al. 2015), anti-
diabetic (Lu et al. 2016), antioxidant (Wang et al. 2013; Zhou et al. 2015), antifungal (Wang
et al. 2009), anti-viral, apoptosis-inducing and other activities (Li et al. 2011). Thus, the phy-
tochemical and pharmacological studies of Polygonatum species are very actually. In our
previous studies described isolation and characterisation of steroidal glycosides, flavonoids,
stilbenes and coumarins from Polygonatum glaberrimum, Polygonatum polyanthemum and
Polygonatum verticillatum (Gvazava and Kikoladze 2011a, 2011b, 2013, 2014; Gvazava and
CONTACT aꢉꢇxꢆnꢃꢇꢅ skꢀiꢅꢁꢉꢆꢃzꢇ
ꢆꢉꢇkꢂꢆnꢃꢅꢇ.ꢂkꢀiꢅꢁꢉꢆꢃzꢇ@yꢆꢀꢄꢄ.ꢈꢄm
sꢊppꢉꢇmꢇnꢁꢆꢉ ꢃꢆꢁꢆ fꢄꢅ ꢁꢀiꢂ ꢆꢅꢁiꢈꢉꢇ ꢈꢆn bꢇ ꢆꢈꢈꢇꢂꢂꢇꢃ ꢆꢁ ꢀꢁꢁpꢂ://ꢃꢄi.ꢄꢅg/10.1080/14786419.2018.1431628.
©
2018 Infꢄꢅmꢆ uK limiꢁꢇꢃ, ꢁꢅꢆꢃing ꢆꢂ tꢆyꢉꢄꢅ & Fꢅꢆnꢈiꢂ Gꢅꢄꢊp

2
L. GVAZAVA ET AL.
Skhirtladze 2016). Polygonatum multiflorum (L.) All. is a perennial herbaceous plant wide-
spread in Georgia. It is used mainly in the form of a poultice and is believed to prevent
excessive bruising and to stimulate tissue repair. The root is astringent, demulcent, emetic
and tonic. An infusion is healing and restorative, it is good in the treatment of stomach
inflammations, chronic dysentery etc. Continuing our investigation of secondary metabolites
in genus Polygonatum, our studies were focused on the isolation and identification of dios-
genin derivative furostanol glycosides from P. multiflorum. Two novel (4, 5) and three known
furostanols were isolated using combined column chromatography with Diaion HP-20,
Sephadex LH-20 and Silica gel, and structurally elucidated by various spectral analysis.
2
. Results and discussion
From the MeOH extract of P. multiflorum five furostanol glycosides (1–5) were isolated by
repeated chromatography on Diaion HP-20, Sephadex LH-20 followed on Silica gel column
to afford two new furostanol glycosides (4, 5), together with three known glycosides: pro-
tobioside (1) (Geng et al. 2004), protodeltonin (2) (Ori et al. 1992), huangjiangsu A (3) (Ali
et al. 2013) (Figure 1). The structures of isolated compounds were established by intensive
1
13
use of mono- and bi-dimensional NMR ( H, C, COSY, HSQC, HMBC), as well as mass-spec-
trometry (ESI/MS).
The molecular formula of 4 was unequivocally established to be C H105O33 by HRTOFMS
6
3
+
(
m/z 1389.4151 [M+H] ). The positive ESI/MS mass spectrum of 4 showed the major ion peak
at m/z 1389, which was assigned to [M+H] . The MS/MS of this ion showed peaks at m/z 1209
M+H−18−162] , due to the loss of water and sugar group, and at m/z 1047 [M+H−18−162×2] ,
m/z 885 [M+H−18−162×3] , m/z 723 [M+H−18−162×4] , m/z 577 [M+H−18−162×4−146] ,
+
+
+
[
+
+
+
+
m/z 415 [M+H−18−162×5−146] , corresponding to the loss of one, two, three hexose, doex
-
yhexose and then another hexose units, respectively.
1
The H NMR spectrum of 4 showed signals for two tertiary methyl groups at δ 0.84 (3H,
H
s, Me-18), 1.05 (3H, s, Me-19), two secondary methyl groups at δ 0.95 (3H, d, J = 6.7 Hz,
H
Me-27), 1.00 (3H, d, J = 6.7 Hz, Me-21), two methine proton signals at δ 3.59 (1H, m, H-3)
H
and 4.37 (1H, m, H-16) indicative of secondary alcoholic functions, two methylene proton
signals at δ 3.73 (1H, dd, J = 10.5, 6.0 Hz, H -26) and 3.44 (1H, m, H -26) ascribable to a pri-
H
a
b
mary alcoholic function, and six anomeric protons at δ 5.25 (1H, br s, H-1″), 4.52 (1H, d,
H
J = 7.8 Hz, H-1′), 4.49 (1H, d, J = 7.8 Hz, H-1″″), 4.48 (1H, d, J = 7.8 Hz, H-1″′), 4.33 (1H, d,
J = 7.7 Hz, H-1″″′) and 4.23 (1H, d, J = 7.9 Hz, H-1″″″), as well as a signal for an oleꢀnic proton
13
at δ 5.38 (1H, br d, J = 4.5 Hz, H-6). The C NMR spectrum displayed signals ascribable to a
H
double bond in position C-5(6) at δ 141.9 (C-5) and 122.3 (C-6), two secondary alcoholic
C
functions at δ 79.2 (C-3) and 82.1 (C-16), and one primary alcoholic function at δ 75.9 (C-26),
C
C
suggesting the occurrence of a glycosidic furostanol skeleton (Table S1). The C-25 configu-
ration was deduced to be R based on the difference of chemical shifts (Δab = δa−δb) of the
geminal protons at H -26 (Δab = 0.29 ppm). It was revealed that Δab was usually >0.57 ppm
a,b
in 25S compounds and <0.48 in 25R compounds (Skhirtladze et al. 2015). The HMBC spectrum
showed correlation peaks between the proton signal at δ 0.84 (3H, s, Me-18) and the carbon
H
resonance at δ 141.9 (C-5), between the proton signal at δ 1.65 (1H, m, H-8) and the carbon
C
H
resonance at δ 122.3 (C-6), confirming a double bond in position C-5(6). On the basis of the
C
HSQC and HMBC correlations, the aglycon moiety of compound 4 was identified as (25R)-
1
13
2
2α-hydroxyfurost-5-en-3β,26-diol (Matsuo et al. 2013). It was evident from the H and
C

NATURAL PRODUCT RESEARCH
3
Figure 1. sꢁꢅꢊꢈꢁꢊꢅꢇꢂ ꢄf ꢈꢄmpꢄꢊnꢃꢂ 1–5.
NMR data that the glycosidic moiety of 4 consisted of six sugar units. The chemical shifts of
all individual protons of the six sugar units were ascertained by a combination of HSQC,
13
HMBC and COSY spectral analysis, and the C NMR chemical shifts of their relative attached
carbons were determined unambiguously from the HSQC spectrum (Table S2). These data
showed the presence of five β-glucopyranosyl units at δ 4.52, 4.49, 4.48, 4.33 and 4.23, and
H

4
L. GVAZAVA ET AL.
one α-rhamnopyranosyl unit at δ 5.25 (Table S2). An unambiguous determination of the
H
sequence and linkage sites was obtained from the HMBC spectrum, which showed key cor-
relation peaks between the proton signal at δ 4.52 (H-1′) and the carbon resonance at δC
H
7
9.2 (C-3), the proton signal at δ 5.25 (H-1″) and the carbon resonance at δ 78.1 (C-2′), the
H
C
proton signal at δ 4.48 (H-1″′) and the carbon resonance at δ 81.0 (C-4′), the proton signal
H
C
at δ 4.49 (H-1″″) and the carbon resonance at δ 88.9 (C-3″′), the proton signal at δ 4.33
H
C
H
(
H-1′″″) and the carbon resonance at δ 70.0 (C-6″″), and the proton signal at δ 4.23 (H-1″″″)
C
H
and the carbon resonance at δ 75.9 (C-26). Based on these data, the structure of compound
C
4
was established as 26-O-β-D-glucopyranosyl (25R), 22α-hydroxyfurost-5-en-3β,26-diol
3-O-β-D-glucopyranosyl-(1→6)-O-β-D-glucopyranosyl-(1→3)-O-β-D-glucopyranosyl-(1→4)-
O-[α-L-rhamnopyranosyl-(1→2)]-O-β-D-glucopyranoside and named polmultoside A.
The molecular formula of 5 was established to be C H O by HRTOFMS (m/z 1371.3613
6
3
103 32
+
[
M+H] ). The positive ESIMS mass spectrum of 5 showed the major ion peak at m/z 1371,
which was assigned to [M+H] . The MS/MS of this ion showed peaks at m/z 1209 [M+H−162] ,
due to the loss of sugar group, and at m/z 1047 [M+H−162×2] , m/z 885 [M+H−162×3] , m/z
23 [M+H−162×4] , m/z 577 [M+H−162×4−146] , m/z 415 [M+H−162×5−146] , correspond-
+
+
+
+
+
+
+
7
ing to the loss of one, two, three hexose, doexyhexose and then another hexose units,
respectively.
1
The H NMR spectrum of 5 showed signals for one secondary methyl group at δ 0.94
H
(
3H, d, J = 6.6 Hz, Me-27) and three tertiary methyl groups at δ 0.73 (3H, s, Me-18), 1.04 (3H,
H
s, Me-19), 1.61 (3H, s, Me-21) indicating the presence of double bond in position C-20(22).
HMBC showed key correlation peaks between the proton signal at δ 1.61 (Me-21) and the
H
1
13
carbon resonances at δ 105.2 (C-20) and 152.8 (C-22). By comparing of H and C NMR
C
spectra (Table S1) with literature data the aglycon of 5 was identified as (25R)-furost-5,20(22)-
dien-3β,26-diol – pseudodiosgenin (Ali et al. 2013). The sugar part of 5 was identical to that
of 4 (Table S2). Thus, the structure of 5 was established as 26-O-β-D-glucopyranosyl (25R)-
furost-5,20(22)-dien-3β,26-diol 3-O-β-D-glucopyranosyl-(1→6)-O-β-D-glucopyranosyl-(1→3)-
O-β-D-glucopyranosyl-(1→4)-O-[α-L-rhamnopyranosyl-(1→2)]-O-β-D-glucopyranoside, and
named polmultoside B.
3
. Experimental
3
.1. General
Optical rotation was recorded on a Perkin-Elmer 192 polarimeter. GC: Agilent 7850B; HP-5
capillary column (28 m × 0.32 mm, i.d.); detection by FID; detector temp. 260 °C; column
temp. 180 °C; carrier gas N ; flow-rate 40 mL/min). IR spectrum was recorded on a Perkin-
2
Elmer 1600 spectrometer. NMR experiments were performed on an Avance II 600 MHz spec-
trometer (BrukerBioSpinGmbH, Rheinstetten, Germany) equipped with a Bruker TXI probe
head at 300 K. All 1D- and 2D NMR spectra were recorded in CD OD (99.95%, Euriso-Top),
3
and standard pulse sequences and phase cycling were used for DQF-COSY, HSQC and HMBC
spectra. The NMR data were processed using MestRe-C UXNMR software (Santiago de
Compostela, Spain). High-resolution masses were recorded on a microOTOF-QII (Bruker)
mass spectrometer in the positive electrospray ionisation (ESI) mode. Exact mass calibration
was performed on daily bases with the ESI-L low concentration tuning mix from Agilent
(
Santa Clara, U.S.A). Column chromatography was performed on Diaion HP-20 material

NATURAL PRODUCT RESEARCH
5
(
Sigma-Aldrich) and silica gel (0.063–0.100 mm, Merck). TLC was performed on silica gel
plates (Merck silica gel 60 F254), which were developed in the solvent system CHCl /MeOH/
3
H O (26:14:3). All solvents for extraction and chromatographic separation were of analytical
2
grade and purchased from Merck (Darmstadt, Germany).
3
.2. Plant material
The whole plant P. multiflorum was collected in July 2015 in Kartli region (Georgia). Samples
of P. multiflorum were identiꢀed by Dr. Jemal Aneli, Department of Pharmacobotany, Institute
of Pharmacochemistry, Tbilisi, Georgia, and a respective herbarium specimen (No. 7242) was
deposited at this department.
3
.3. Extraction and isolation
500 g of powdered P. multiflorum was extracted with 2.5 l of 80% MeOH for 1 h three times
at 60 °C. The collected extracts were dried under reduced pressure (27 g), and the concentrate
was partitioned between CHCl (6 g) and H O (21 g). A part of the H O extract (10 g) was
3
2
2
subjected to Diaion HP-20 column chromatography (50 × 4 cm) and eluted with a gradient
system of H O-MeOH 10:0 to 0:10 to yield four fractions (500 mL each): H O (2.3 g), 30%
2
2
MeOH (2.9 g), 80% MeOH (4.2 g) and MeOH (0.6 g). A part of the 80% MeOH fraction (3 g)
was separated on Sephadex LH-20 (100 g), eluted with MeOH, enriched fractions (1–6 and
7
–16) were then separated by CC (SiO ; 100 g, 500 × 25 mm), eluted with CHCl /MeOH/H O
2
3
2
26:14:3, to afford 5 individual furostanol glycosides. From the fractions 1–6 compounds 1
(
(
5.6 mg), 2 (7.6 mg), 3 (7.8 mg) and from the fractions 7–16 compounds 4 (7.4 mg) and 5
12.9 mg) were obtained.
Polmultoside A (4): 26-O-β-D-glucopyranosyl (25R), 22α-hydroxyfurost-5-en-3β,26-diol
3
-O-β-D-glucopyranosyl-(1→6)-O-β-D-glucopyranosyl-(1→3)-O-β-D-glucopyranosyl-(1→4)-
20
O-[α-L-rhamnopyranosyl-(1→2)]-O-β-D-glucopyranoside. White amorphous powder; [ꢀ]D :
58.3 (c 0.59, pyridine); Rf = 0.41 (solvent system: CHCl /MeOH/H O (26:14:3); IR ν (KBr)
−
3
1 1
2
max
−
3400 (OH), 2925 (CH), 1642, 1459, 1069, 920, 890 cm ; H NMR (600 MHz, CD OD) δ 1.88
3 H
(1H, m, Ha-1), 1.08 (1H, m, Hb-1), 1.92 (1H, m, Ha-2), 1.61 (1H, m, Hb-2), 3.59 (1H, m, H-3), 2.45
(
1H, dd, J = 13.0, 2.5 Hz, Ha-4), 2.30 (1H, t, J = 13.0 Hz, Hb-4), 5.38 (1H, br d, J = 4.5 Hz, H-6),
2
1
.01 (1H, m, Ha-7), 1.55 (1H, m, Hb-7), 1.65 (1H, m, H-8), 0.97 (1H, m, H-9), 1.55 (2H, m, Ha,b-
1), 1.77 (1H, m, Ha-12), 1.19 (1H, m, Hb-12), 1.13 (1H, m, H-14), 1.97 (1H, m, Ha-15), 1.28 (1H,
m, Hb-15), 4.37 (1H, m, H-16), 1.73 (1H, dd, J = 8.0, 6.5 Hz, H-17), 0.84 (3H, s, Me-18), 1.05 (3H,
s, Me-19), 2.18 (1H, m, H-20), 1.00 (3H, d, J = 6.7 Hz, Me-21), 1.82 (1H, m, Ha-23), 1.63 (1H, m,
Hb-23), 1.59 (1H, m, Ha-24), 1.16 (1H, m, Hb-24), 1.76 (1H, m, H-25), 3.73 (1H, dd, J = 10.5,
6
3
.0 Hz, Ha-26), 3.44 (1H, m, Hb-26), 0.95 (3H, d, J = 6.7 Hz, Me-27), 4.52 (1H, d, J = 7.8 Hz, H-1′),
.42 (1H, dd, J = 9.0, 7.8 Hz, H-2′), 3.66 (1H, dd, J = 9.0, 9.0 Hz, H-3′), 3.56 (1H, dd, J = 9.0, 9.0 Hz,
H-4′), 3.38 (1H, m, H-5′), 3.83 (1H, dd, J = 11.6, 2.4 Hz, Ha-6′), 3.66 (1H, dd, J = 11.6, 4.8 Hz,
Hb-6′), 5.25 (1H, br s, H-1″), 3.89 (1H, br d, J = 1.7 Hz, H-2″), 3.66 (1H, dd, J = 9.2, 3.0 Hz, H-3″),
3
4
.40 (1H, t, J = 9.1 Hz, H-4″), 4.12 (1H, dd, J = 9.0, 6.2 Hz, H-5″), 1.24 (1H, d, J = 6.2 Hz, H-6″),
.48 (1H, d, J = 7.8 Hz, H-1‴), 3.40 (1H, dd, J = 8.8, 7.8 Hz, H-2‴), 3.55 (1H, dd, J = 9.0, 8.8 Hz,
H-3‴), 3.44 (1H, dd, J = 8.8, 8.8 Hz, H-4‴), 3.38 (1H, m, H-5‴), 3.88 (1H, dd, J = 11.0, 2.1 Hz,
Ha-6‴), 3.65 (1H, dd, J = 11.0, 5.0 Hz, Hb-6‴), 4.49 (1H, d, J = 7.8 Hz, H-1″″), 3.30 (1H, dd, J = 9.0,
7.8 Hz, H-2″″), 3.27 (1H, dd, J = 9.0, 9.0 Hz, H-3″″), 3.29 (1H, dd, J = 9.0, 9.0 Hz, H-4″″), 3.58 (1H,

6
L. GVAZAVA ET AL.
m, H-5″″), 4.27 (1H, dd, J = 11.8, 2.1 Hz, Ha-6″″), 3.64 (1H, dd, J = 11.8, 4.8 Hz, Hb-6″″), 4.33
1H, d, J = 7.7 Hz, H-1″″′), 3.20 (1H, dd, J = 8.8, 7.7 Hz, H-2″″′), 3.25 (1H, dd, J = 9.0, 8.8 Hz,
(
H-3″″′), 3.28 (1H, dd, J = 9.0, 9.0 Hz, H-4″″′), 3.32 (1H, m, H-5″″′), 3.88 (1H, br d, J = 11.0, 2.5 Hz,
Ha-6″″′), 3.67 (1H, dd, J = 11.0, 4.9 Hz, Hb-6″″′), 4.23 (1H, d, J = 7.9 Hz, H-1″″″), 3.18 (1H, dd,
J = 9.0, 7.9 Hz, H-2″″″), 3.35 (1H, dd, J = 9.0, 9.0 Hz, H-3″″″), 3.29 (1H, dd, J = 9.0, 9.0 Hz, H-4″″″),
3
.26 (1H, m, H-5″″″), 3.84 (1H, dd, J = 11.6, 2.4 Hz, Ha-6″″″), 3.67 (1H, dd, J = 11.6, 4.8 Hz,
13
Hb-6″″″); C NMR (300 MHz, CD OD) δ 38.4 (C-1), 30.5 (C-2), 79.2 (C-3), 39.2 (C-4), 141.9 (C-5),
3
C
122.3 (C-6), 33.0 (C-7), 32.6 (C-8), 51.7 (C-9), 37.3 (C-10), 21.8 (C-11), 40.7 (C-12), 41.4 (C-13),
57.3 (C-14), 32.5 (C-15), 82.1 (C-16), 64.5 (C-17), 16.6 (C-18), 19.7 (C-19), 40.9 (C-20), 15.9 (C-21),
111.7 (C-22), 31.2 (C-23), 28.8 (C-24), 34.8 (C-25), 75.9 (C-26), 17.1 (C-27), 100.1 (C-1′), 78.1
(
(
C-2′), 77.6 (C-3′), 81.0 (C-4′), 76.9 (C-5′), 61.9 (C-6′), 101.7 (C-1″), 72.0 (C-2″), 72.2 (C-3″), 73.7
C-4″), 69.4 (C-5″), 17.8 (C-6″), 104.3 (C-1″′), 73.8 (C-2″′), 88.9 (C-3″′), 69.6 (C-4″′), 77.4 (C-5″′),
6
1.8 (C-6″′), 104.4 (C-1″″), 75.0 (C-2″″), 77.8 (C-3″″), 71.4 (C-4″″), 76.5 (C-5″″), 70.0 (C-6″″), 104.0
(
(
(
C-1″″′), 75.1 (C-2″″′), 77.7 (C-3″″′), 71.5 (C-4″″′), 77.8 (C-5″″′), 62.0 (C-6″″′), 104.3 (C-1″″″), 75.0
C-2″″″), 77.8 (C-3″″″), 71.4 (C-4″″″), 77.7 (C-5″″″), 62.3 (C-6″″″); HR-ESI-TOF-MS: 1389.4151
+
+
[M+H] , C H104O33 ; calcd for 1389.4139).
6
3
Polmultoside B (5): 26-O-β-D-glucopyranosyl (25R)-furost-5,20(22)-dien-3β,26-diol 3-O-β-
D-glucopyranosyl-(1→6)-O-β-D-glucopyranosyl-(1→3)-O-β-D-glucopyranosyl-(1→4)-O-[α-L-
2
0
rhamnopyranosyl-(1→2)]-O-β-D-glucopyranoside. White amorphous powder;[ꢀ]D : −45.8 (c
0
2
.62, pyridine); Rf = 0.49 (solvent system: CHCl /MeOH/H O (26:14:3); IR ν (KBr) 3412 (OH),
3 2 max
−1 1
923 (CH), 1644, 1460, 1069, 893 cm ; H NMR (600 MHz, CD OD) δ 1.87 (1H, m, Ha-1), 1.08
3
H
(
1H, m, Hb-1), 1.92 (1H, m, Ha-2), 1.61 (1H, m, Hb-2), 3.58 (1H, m, H-3), 2.44 (1H, dd, J = 13.0,
2
1
.5 Hz, Ha-4), 2.30 (1H, t, J = 13.0 Hz, Hb-4), 5.39 (1H, br d, J = 4.5 Hz, H-6), 1.99 (1H, m, Ha-7),
.55 (1H, m, Hb-7), 1.64 (1H, m, H-8), 0.97 (1H, m, H-9), 1.54 (2H, m, Ha,b-11), 1.78 (1H, m,
Ha-12), 1.19 (1H, m, Hb-12), 1.01 (1H, m, H-14), 2.17 (1H, m, Ha-15), 1.41 (1H, m, Hb-15), 4. 73
1H, m, H-16), 2.50 (1H, d, J = 10.0 Hz, H-17), 0.73 (3H, s, Me-18), 1.04 (3H, s, Me-19), 1.61 (3H,
s, Me-21), 2.13 (2H, m, Ha,b-23), 1.63 (1H, m, Ha-24), 1.26 (1H, m, Hb-24), 1.78 (1H, m, H-25),
.75 (1H, dd, J = 10.5, 6.0 Hz, Ha-26), 3.43 (1H, m, Hb-26), 0.94 (3H, d, J = 6.7 Hz, Me-27), 4.52
1H, d, J = 7.8 Hz, H-1′), 3.42 (1H, dd, J = 9.0, 7.8 Hz, H-2′), 3.66 (1H, dd, J = 9.0, 9.0 Hz, H-3′),
.56 (1H, dd, J = 9.0, 9.0 Hz, H-4′), 3.38 (1H, m, H-5′), 3.83 (1H, dd, J = 11.6, 2.4 Hz, Ha-6′), 3.66
1H, dd, J = 11.6, 4.8 Hz, Hb-6′), 5.25 (1H, br s, H-1″), 3.89 (1H, br d, J = 1.7 Hz, H-2″), 3.66 (1H,
dd, J = 9.2, 3.0 Hz, H-3″), 3.40 (1H, t, J = 9.1 Hz, H-4″), 4.12 (1H, dd, J = 9.0, 6.2 Hz, H-5″), 1.24
(
3
(
3
(
(
(
1H, d, J = 6.2 Hz, H-6″), 4.48 (1H, d, J = 7.8 Hz, H-1″′), 3.40 (1H, dd, J = 8.8, 7.8 Hz, H-2″′), 3.55
1H, dd, J = 9.0, 8.8 Hz, H-3″′), 3.44 (1H, dd, J = 8.8, 8.8 Hz, H-4″′), 3.38 (1H, m, H-5″′), 3.88 (1H,
dd, J = 11.0, 2.1 Hz, Ha-6″′), 3.65 (1H, dd, J = 11.0, 5.0 Hz, Hb-6″′), 4.49 (1H, d, J = 7.8 Hz, H-1″″),
3.30 (1H, dd, J = 9.0, 7.8 Hz, H-2″″), 3.27 (1H, dd, J = 9.0, 9.0 Hz, H-3″″), 3.29 (1H, dd, J = 9.0,
9.0 Hz, H-4″″), 3.58 (1H, m, H-5″″), 4.27 (1H, dd, J = 11.8, 2.1 Hz, Ha-6″″), 3.64 (1H, dd, J = 11.8,
4.8 Hz, Hb-6″″), 4.33 (1H, d, J = 7.7 Hz, H-1″″′), 3.20 (1H, dd, J = 8.8, 7.7 Hz, H-2″″′), 3.25 (1H,
dd, J = 9.0, 8.8 Hz, H-3″″′), 3.28 (1H, dd, J = 9.0, 9.0 Hz, H-4″″′), 3.32 (1H, m, H-5″″′), 3.88 (1H,
br d, J = 11.0, 2.5 Hz, Ha-6″″′), 3.67 (1H, dd, J = 11.0, 4.9 Hz, Hb-6″″′), 4.23 (1H, d, J = 7.9 Hz,
H-1″″″), 3.18 (1H, dd, J = 9.0, 7.9 Hz, H-2″″″), 3.35 (1H, dd, J = 9.0, 9.0 Hz, H-3″″″), 3.29 (1H, dd,
J = 9.0, 9.0 Hz, H-4″″″), 3.26 (1H, m, H-5″″″), 3.84 (1H, dd, J = 11.6, 2.4 Hz, Ha-6″″″), 3.67 (1H,
dd, J = 11.6, 4.8 Hz, Hb-6″″″); 13C NMR (300 MHz, CD OD) δ 38.4 (C-1), 30.4 (C-2), 79.1 (C-3),
3
C
39.3 (C-4), 142.0 (C-5), 122.3 (C-6), 32.9 (C-7), 32.6 (C-8), 51.6 (C-9), 37.4 (C-10), 21.9 (C-11),
40.5 (C-12), 44.2 (C-13), 56.2 (C-14), 34.8 (C-15), 85.4 (C-16), 65.3 (C-17), 14.1 (C-18), 19.7 (C-19),
105.2 (C-20), 11.8 (C-21), 152.8 (C-22), 24.0 (C-23), 31.8 (C-24), 34.0 (C-25), 75.8 (C-26), 17.2

NATURAL PRODUCT RESEARCH
7
(
(
C-27), 100.1 (C-1′), 78.1 (C-2′), 77.6 (C-3′), 81.0 (C-4′), 76.9 (C-5′), 61.9 (C-6′), 101.7 (C-1″), 72.0
C-2″), 72.2 (C-3″), 73.7 (C-4″), 69.4 (C-5″), 17.8 (C-6″), 104.3 (C-1″′), 73.8 (C-2″′), 88.9 (C-3″′),
69.6 (C-4″′), 77.4 (C-5″′), 61.8 (C-6″′), 104.4 (C-1″″), 75.0 (C-2″″), 77.8 (C-3″″), 71.4 (C-4″″), 76.5
(
(
C-5″″), 70.0 (C-6″″), 104.0 (C-1″″′), 75.1 (C-2″″′), 77.7 (C-3″″′), 71.5 (C-4″″′), 77.8 (C-5″″′), 62.0
C-6″″′), 104.3 (C-1″″″), 75.0 (C-2″″″), 77.8 (C-3″″″), 71.4 (C-4″″″), 77.7 (C-5″″″), 62.3 (C-6″″″);
+
+
HR-ESI-TOF-MS: 1371.3613 ([M+H] , C H102O32 ; calcd for 1371.3620).
6
3
3
.4. Acid hydrolysis
Compounds 4 and 5 (5–5 mg) were dissolved in aqueous MeOH (50%, 2 mL) containing
conc. H SO (0.2 mL), refluxed for 8 h, cooled and diluted with H O. The resulting precipitates
2
4
2
were filtered off and purified by recrystallization from MeOH to afford white crystals
1.8–2 mg, respectively) of the aglycons. The aglycons were identified as (25R)-spirrost-5-en-
β-ol (diosgenin) by comparing the NMR data with the literature. The aqueous layers were
(
3
neutralised with dilute NaOH and concentrated to dryness. The resultant residues were dis-
solved in pyridine (1 mL), then (CH ) SiNHSi (CH ) (1 mL) was added. After 10 min at room
3
3
3 3
temperature, the solutions were blown to dryness under a stream of nitrogen. The residues
were dissolved in diethyl ether and then subjected to chiral GC-MS analysis. Absolute
conꢀgurations of monosaccharides were found to be D-glucose and L-rhamnose by com-
parison of the retention times (T ) of their derivatives with those of L-rhamnose (T = 4.25 min)
R
R
and D-glucose (T = 10.53 min) derivatives prepared in the same way.
R
4
. Conclusion
Two new steroidal glycosides, polmultoside A (4) and polmultoside B (5), along with three
known glycosides protobioside (1), protodeltonin (2) and huangjiangsu A (3) were isolated
from P. multiflorum (L.) All. P. multiflorum can be considered as a rich source of steroidal
glycosides which characteristically shows Δ5 rings A and B, belonging to the diosgenin
furostanols.
Disclosure statement
No potential conflict of interest was reported by the authors.
References
Ali Z, Smillie TJ, Khan IA. 2013. Cholestane steroid glycosides from the rhizomes of Dioscorea villosa
(wild yam). Carbohydrate res. 370:86–91.
Gagnidze R. 2011. Flora Georgia. 2nd ed. Vol. XVI. Universaly, Tbilisi, Georgia.
Geng Y, Tan NH, Jun Z, Kong LY. 2004. Isolation and identification of steroid saponins from the fresh
rhizomes of Dioscorea panthaica. Chin J Nat Med. 2:25–27.
Gvazava LN, Kikoladze VS. 2011a. Flavonoids from the plants Polygonatum polyanthemum and P.
glaberimum. Chem Nat Compd. 47:818–819.
Gvazava LN, Kikoladze VS. 2011b. Stilbenes and coumarins from the plant Polygonatum polyanthemum.
Chem Nat Compd. 47:810–811.
Gvazava LN, Kikoladze VS. 2013. Steroidal saponins from Polygonatum polyanthemum rhizomes. Chem
Nat Compd. 49:173–174.

8
L. GVAZAVA ET AL.
Gvazava LN, Kikoladze VS. 2014. Steroidal saponins from Polygonatum glaberimum rhizomes. Chem
Nat Compd. 50:489–493.
Gvazava LN, Skhirtladze AV. 2016. Steroidal saponins from Polygonatum verticillatum. Chem Nat Compd.
52:1052–1055.
Kazmi I, Afzal M, Rahman M, Gupta G, Anwar F. 2012. Aphrodisiac properties of Polygonatum verticillatum
leaf extract. Asian Pac J Trop Diss. 2:S841–S845.
Khan H, Saeed M, Rauf A, Khan MA, Muhammad N. 2015. Antimicrobial and inhibition on heat-induced
protein denaturation of constituents isolated from Polygonatum verticillatum rhizomes. Nat Prod
Res. 29(22):2160–2163.
Li CY, Luo P, Liu JJ, Wang EQ, Li WW, Ding ZH, Mou L, Bao JK. 2011. Recombinant expression of
Polygonatum cytonema lectin with anti-viral, apoptosis-inducing and preliminary crystallization.
Proc Biochem. 46:533–542.
Lu JM, Wang YF, Yan HL, Lin P, Gu W. 2016. Antidiabetic effect of total saponins from Polygonatum
kingianum in streptozotocin-induced diabetic rats. J Ethnopharmacology. 179:291–300.
Matsuo Y, Shinoda D, Nakamaru A, Mimaki Y. 2013. Steroidal glycosides from the bulbs of Fritillaria
meleagris and their cytotoxic activities. Steroids. 78:670–682.
Ori K, Mimaki Y, Mito K, Sashida Y, Nikaido T, Ohmoto T, Masuko A. 1992. Jatropham derivatives and
steroidal saponins from the bulbs of Lilium hansonii. Phytochem. 31:2767–2775.
Skhirtladze A, Benidze M, Kemertelidze E, Grigolava B, Sturm S, Ganzera M. 2015. Steroid composition
of fruit from Yucca gloriosa introduced into Georgia. Chem Nat Compd. 51:283–288.
Wang D, Li D, Zhu W, Zhang J, Peng P. 2009. Steroidal saponins from the rhizomes of Polygonatum
odoratum. Nat Prod Res. 23(10):940–947.
Wang D, Zeng L, Li D, Pu W. 2013. Antioxidant activities of different extracts and homoisoflavanones
isolated from the Polygonatum odoratum. Nat Prod Res. 27(12):1111–1114.
Zhou X, Yuping Z, Zhao H, Liang J, Zhang Y. 2015. Antioxidant homoisoflavonoids from Polygonatum
odoratum. Food chem. 186:63–68.