Two new triterpenoid saponins from Pittosporum senacia Putterlick (Pittosporaceae)

Article abstract of DOI:10.1002/mrc.3876

From the branches of Pittosporum senacia Putterlick (Pittosporaceae), two new triterpenoid saponins, senaciapittosides A and B (1, 2), were isolated. Their structures were elucidated by extensive analysis of one- and two-dimensional nuclear magnetic reson

Full text of DOI:10.1002/mrc.3876

MRC Letters
Received: 6 April 2012
Revised: 5 July 2012
Accepted: 9 July 2012
Published online in Wiley Online Library
(wileyonlinelibrary.com) DOI 10.1002/mrc.3876
Two new triterpenoid saponins from
Pittosporum senacia Putterlick (Pittosporaceae)
Jens Linnek,a Anne-Claire Mitaine-Offer,a Thomas Paululatb and
Marie-Aleth Lacaille-Duboisa*
From the branches of Pittosporum senacia Putterlick (Pittosporaceae), two new triterpenoid saponins, senaciapittosides A and
B (1, 2), were isolated. Their structures were elucidated by extensive analysis of one- and two-dimensional nuclear magnetic
resonance spectroscopy, high-resolution electrospray ionization mass spectrometry (HR-ESIMS) and chemical evidence as
3-O-[b-D-glucopyranosyl-(1 ! 2)]-[a-L-arabinopyranosyl-(1! 3)]-[a-L-arabinofuranosyl-(1 ! 4)]-b-D-glucuronopyranosyl oleanolic
acid 28-O-b-D-glucopyranosyl ester (1) and 3-O-[b-D-glucopyranosyl-(1 ! 2)]-[a-L-arabinopyranosyl-(1 ! 3)]-[a-L-arabinofuranosyl-
(1 ! 4)]-b-D-glucuronopyranosyl-22-O-a-L-arabinopyranosyl-21-acetoxy R1-barrigenol (2). Compound 2 presents an unusual
glycosylation at C-22 of its aglycone. Copyright © 2012 John Wiley & Sons, Ltd.
1
Keywords: NMR; H; 13C; 2D NMR; triterpene saponins; Pittosporaceae; Pittosporum senacia
1220. Another fragment ion was observed at 1087 [(M–H)–132]ꢀ
Introduction
corresponding to the loss of one pentosyl moiety.
The 1H NMR spectrum of the aglycone part of 1 showed seven
The Pittosporaceae family is represented by nine genera,
which contains approximately 250 species, distributed in tropical
and subtropical regions.[1] The use for local traditional medicine
from species of the Pittosporaceae family has been reported in
multiple cases.[2–5] The presence of triterpene saponins in the
genus Pittosporum has been previously reported.[6–8] Some
different endemic subspecies of Pittosporum senacia are known
to have an indigenous medicinal role in tropical regions for
treatment of rheumatism and throat infections.[4,5] A previous work
on the volatile constituents of the leaves of P. senacia var. coursii
Cufodontis was reported,[9] whereas triterpene saponins have
never been characterized in this species. In this article, we report
about the isolation and characterization of two new saponins
from P. senacia Putterlick. The structures were elucidated mainly
by a 600-MHz nuclear magnetic resonance (NMR) analysis including
one-dimensional (1D) and two-dimensional (2D) NMR (1H, 13C NMR,
COSY, TOCSY, ROESY, HSQC and HMBC) spectroscopy and mass
spectrometry.
tertiary methyl groups as singlets at dH 0.69, 0.75, 0.86, 0.87, 0.88,
0.97 and 1.08, one olefinic proton at dH 5.16 (br s, H-12) and one
oxygen bearing methine proton at dH 2.99 (H-3), characteristic of
a triterpene skeleton. Furthermore, the shielded chemical shift in
the 13C NMR spectrum at dC 175.1 (C-28) in comparison with a free
carboxyl group (dC 180.0) suggested an ester function. The
extensive analysis of 1D and 2D NMR spectra (1H, 13C NMR, ROESY,
HSQC and HMBC) led to the identification of the aglycone part of 1
as oleanolic acid (=3-b-hydroxy-olean-12-en-28-oic acid) and was in
1
full agreement with literature data.[10] The H NMR spectrum of 1
showed five anomeric proton signals at dH 5.24 (d, J = 8.1 Hz), 5.06
(br s), 4.82 (d, J = 6.9Hz), 4.71 (d, J = 7.3Hz) and 4.28 (d, J = 6.9Hz),
which correlated in the HSQC spectrum with five anomeric carbons
at dC 94.0, 107.1, 101.7, 101.3 and 103.6, respectively, and indicated
the presence of five sugar moieties. The bidesmosidic structure
has been revealed by the chemical shift of C-3 (dC 88.4) and C-28
(dC 175.1). Complete assignments of each glycosidic proton
system were achieved by COSY, TOCSY and ROESY experiments
starting from the readily identifiable anomeric protons. Thus, one
b-glucuronopyranosyl acid (GlcA), two b-glucopyranosyl (Glc I and
Glc II), one a-arabinopyranosyl (Ara) and one a-arabinofuranosyl
(Araf) units were identified (see Table 2). The evidence of the sugar
linkages to the aglycone at C-28 and C-3 was in particular given by
Results and Discussion
The dried and powdered branches of P. senacia Putterlick
were refluxed in MeOH. After evaporation of the solvent, the dried
MeOH extract was submitted to several liquid chromatographic
steps [vacuum liquid chromatography (VLC), medium pressure
liquid chromatography (MPLC)] on normal-(SiO2) and reversed-
phase (RP-18 SiO2) affording two new triterpene saponins,
senaciapittosides A (1) and B (2).
*
Correspondence to: Marie-Aleth Lacaille-Dubois, EA 4267, FDE/UFC, Laboratoire
de Pharmacognosie, Faculté de Pharmacie, Université de Bourgogne, 7 bd. Jeanne
D’ Arc, BP 87900, 21079 Dijon cedex, France. E-mail: m-a.lacaille-dubois@u-
bourgogne.fr
The high-resolution electrospray ionization mass spectrometry
(HR-ESIMS, positive-ion mode) spectrum of 1 showed a pseudo-
molecular ion peak at m/z 1243.5729 [M + Na]+ (calculated
1243.5724) ascribable to the molecular formula C58H92O27. The
ESIMS spectrum (negative ion-mode) showed a quasi-molecular
ion peak at m/z 1219 [M–H]ꢀ, indicating a molecular weight of
a EA 4267, FDE/UFC, Laboratoire de Pharmacognosie, Faculté de Pharmacie,
Université de Bourgogne, 7 bd. Jeanne d’Arc, BP 87900, 21079, Dijon cedex,
France
b Organische Chemie II, Naturwissenschaftlich-Technische Fakultät, University of
Siegen, Adolf-Reichwein-Strasse 2, D-57068, Siegen, Germany
Magn. Reson. Chem. (2012)
Copyright © 2012 John Wiley & Sons, Ltd.

J. Linnek et al.
O
O
the HMBC cross peaks dH 5.24 (d, J= 8.1Hz, Glc II H-1)/dC 175.1 (Agly
C-28) and dH 4.28 (d, J = 6.9Hz, GlcA H-1)/dC 88.4 (Agly C-3),
respectively. Furthermore, the linkage of GlcA to the aglycone
at C-3 was confirmed by the ROESY cross peak at dH 4.28
(d, J = 6.9 Hz, GlcA H-1)/dH 2.99 (Agly H-3). The sequence of the
3-O-oligosaccharidic moiety was assigned by HMBC and ROESY
experiments. HMBC cross peaks at dH 3.73 (GlcA H-2)/dC 101.3
(Glc I C-1) and at dH 3.68 (GlcA H-3)/dC 101.7 (Ara C-1) suggested
a Glc I-(1! 2)-[Ara-(1 ! 3)]-GlcA sequence. This was confirmed by
the ROESY cross peaks at dH 4.71 (d, J = 7.3 Hz, Glc I H-1)/dH 3.73
(GlcA H-2) and dH 4.82 (d, J = 6.9Hz, Ara H-1)/dH 3.68 (GlcA H-3).
A strong ROESY cross peak dH 5.06 (br s, Araf H-1)/dH 3.55 (GlcA
H-4) showed the additional Araf-(1 ! 4)-GlcA linkage. The absolute
configurations of the sugars were determined to be D for glucose
and glucuronic acid and L for arabinose by gas chromatography
(GC) analysis (see Experimental section). On the basis of the
previous results, the structure of compound 1 was elucidated
as 3-O-[b-D-glucopyranosyl-(1 ! 2)]-[a-L-arabinopyranosyl-(1 ! 3)]-
[a-L-arabinofuranosyl-(1 ! 4)]-b-D-glucuronopyranosyl oleanolic acid
28-O-b-D-glucopyranosyl ester, named senaciapittoside A (Fig. 1).
Compound 2 was obtained as a white amorphous powder.
The molecular formula of compound 2 was deduced as C59H94O30
on the basis of HR-ESIMS (positive-ion mode) exhibiting the
pseudo-molecular peak at m/z 1305.5731 [M + Na]+ (calculated
1305.5728). The ESIMS (negative-ion mode) showed a quasi-
molecular ion peak at m/z 1281 [M–H]ꢀ, indicating a molecular
weight of 1282. The 1H NMR spectrum corresponding to the
aglycone part of 2 showed as for compound 1 seven methyl groups
as singlets at dH 0.74, 0.76, 0.88, 0.90, 0.91, 0.98 and 1.27 and
one olefinic proton at dH 5.32 (1H, br s, H-12), characteristic of an
oleanene-type skeleton. Furthermore, the presence of five oxygen
bearing methine protons at dH 3.02 (1H, H-3), 3.36 (1H, H-16),
3.58 (1H, H-15), 4.03 (1H, H-22) and 5.48 (1H, d J= 9.3 Hz, H-21),
and one primary alcoholic function at dH 3.12, 3.33 (2 H, H2-28)
suggested a hexahydroxy-3,15,16,21,22,28-oleanene-type skeleton.
Cross peaks in the HMBC spectrum at dH 5.48 (1H, d J= 9.3Hz, H-21)/
dC 19.6 (C-30), 29.1 (C-29), 35.1 (C-20), 78.5 (C-22) and 170.3, respec-
tively (Fig. 2), allowed us to determine the secondary alcoholic
function to be esterified at C-21 by an acetyl group. Furthermore,
the COSY cross peak at dH 5.48 (1H, d J = 9.3Hz, H-21)/dH 4.03
(1H, H-22) assigned unambiguously the location of a further
30
29
21
22
H
H
HMBC
ROESY
COSY
OH
28
O
16
OH
OH
O
HO
OH
Ara II
Figure 2. Correlations of the aglycone part of compound 2 characterized
by HMBC, ROESY and COSY.
secondary alcoholic function at C-22. The HMBC connectivities
dH 4.03 (1H, H-22)/dC 78.3 (C-21), 71.8 (C-16), 61.7 (C-28) and 47.8
(C-17) (Fig. 2) and HMBCs between the protons of the methyl group
at dH 1.27 (3 H, s, H3-27) and the carbon at dC 66.1 (C-15) confirmed
the position of the secondary alcoholic functions at C-15 and C-16.
The stereochemistry was verified by ROESY experiments. Namely,
correlations between dH 3.02 (1H, H-3) and 0.72 (1H, H-5) and dH
0.98 (3H, s, H3-23), between 3.58 (1H, H-15) and 0.91 (3H, s, H3-
26) and between 3.36 (1H, H-16) and 3.12 (1H, H2-28) suggested
the absolute configuration of the R1-barrigenol at C-3, C-15 and
C-16.[6] Furthermore, ROESY cross peaks at dH 0.74 (3H, s, H3-29)/
5.48 (1H, d J = 9.3 Hz, H-21) and 0.88 (3 H, s, H3-30)/4.03 (1H, H-22)
revealed the b-axial orientation of the H-22, and the multiplicity
as a doublet and the coupling constant of 9.3Hz suggested an
a-axial orientation of H-21. This extensive 2D NMR analysis
confirmed the identification of the aglycone of compound 2 as
the already known sapogenol called R1-barrigenol (olean-12-ene-
3b,15a,16a,21b,22a,28-hexol),[6,7,11–16] which is esterified at C-21
30
29
12
26
25
Araf
28
O
OH
GlcA
HO
OH
27
3
O
HOOC
O
O
O
HO
OH
HO
HO
O
O
O
OH
24
23
O
Glc II
HO
O
OH
OH
Ara
O
Glc I
HO
HO
OH
Figure 1. Structure of compound 1.
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Copyright © 2012 John Wiley & Sons, Ltd.
Magn. Reson. Chem. (2012)

Saponins from Pittosporum senacia
1
by an acetyl function. In the H NMR spectrum of compound 2,
Experimental
signals of five anomeric protons were shown at dH 5.06 (br s), 4.81
(d, J = 6.0Hz), 4.72 (d, J =7.3 Hz), 4.28 (d, J = 6.9Hz) and 4.14
(d, J = 5.7Hz), which correlated in the HSQC spectrum with five
carbons at dC 106.8, 101.9, 101.4, 103.6 and 104.6, respectively,
indicating the presence of five sugar moieties. Complete assign-
ments were achieved by 1D and 2D NMR spectra. The evaluation
of chemical shifts and spin-spin couplings allowed the identification
of one b-glucuronopyranosyl acid (GlcA), one b-glucopyranosyl (Glc),
two a-arabinopyranosyl (Ara I and Ara II) and one a-arabinofuranosyl
(Araf ) units (see Table 2). As for compound 1, we have here to deal
with a bidesmosidic structure, showing the sugar linkages to the
aglycone at C-22 and C-3, which were given by the HMBC cross
peaks at dH 4.14 (d, J= 5.7 Hz, Ara II H-1)/dC 78.5 (Agly C-22) and
dH 4.28 (d, J=6.9Hz, GlcA H-1)/dC 88.3 (Agly C-3), respectively.
Furthermore, the linkages of GlcA to C-3 and Ara II to C-22 were
confirmed by the ROESY cross peaks at dH 4.28 (d, J= 6.9Hz, GlcA
H-1)/dH 3.02 (Agly H-3) and dH 4.14 (d, J= 5.7 Hz, Ara II H-1)/dH 4.03
(d, J= 8.5 Hz, Agly H-22). The sugar sequence investigated by exten-
sive 2D NMR experiments revealed that the 1H and 13C NMR signals
of the oligosaccharide chain at C-3 were almost superimposable with
those of (3-O-[Glc-(1 ! 2)]-[Ara-(1 ! 3)]-[Araf-(1 ! 4)]-GlcA) charac-
terized in compound 1. The absolute configurations of the sugars
were determined to be D for glucose and glucuronic acid and L for
arabinose by GC analysis (see Experimental section). Throughout
this information, the structure of compound 2 was elucidated as
3-O-[b-D-glucopyranosyl-(1 ! 2)]-[a-L-arabinopyranosyl-(1 ! 3)]-[a-L-
arabinofuranosyl-(1 ! 4)]-b-D-glucuronopyranosyl-22-O-a-L-
arabinopyranosyl-21-acetoxy R1-barrigenol, named senaciapittoside
B (Fig. 3).
General
The materials and methods used were as follows: VLC, silica gel
60 (15–40mm; Merck); MPLC, silica gel RP-18 Spherical C18
(300Å, 75–200 mm; Silicycle); Pump Module C-601 (Büchi); fraction
collector, C-660 (Büchi); Büchi glass column (460ꢁ 25 mm and
460 ꢁ 15 mm); Büchi precolumn (110 ꢁ 15 mm); rotary evaporator,
Rotavapor R-210 (Büchi); GC, Termoquest gas chromatograph;
DB-1701 capillary column (30 m ꢁ 0.25mm) (J & W Scientific);
detection, Free Induction Decay (FID); detector temperature, 80 ꢂC
for 5 min and raised to 270 ꢂC at the at 15 ꢂC/min; carrier gas, He;
TLC, silica plate F-254 (60 Å, Silicycle); HPTLC, silica gel 60 F254
(Merck); solvent systems, CHCl3/MeOH/H2O 65:32:6.5; detection
with vanillin reagent, that is, 1% 4-hydroxy-3-methoxybenzaldehyd
in EtOH with 2% of a 96% H2SO4, followed by heating; optical rota-
tion, AA-OR automatic polarimeter. NMR spectra were measured on
a Varian VNMR-S 600 MHz spectrometer equipped with 3 mm triple
resonance inverse and 3 mm dual broadband probe heads. Spectra
are recorded in 150ml DMSO-d6. Solvent signals were used as inter-
nal standard (DMSO-d6: dH = 2.50, dC = 39.5 ppm); all spectra are
recorded at T = 35 ꢂC. Pulse sequences were taken from Varian
pulse sequence library and used with following conditions
(compounds 1 and 2): relaxation delay in 2D spectra have been
1.5 s (1) or 2 s (2); gCOSY with 1 (1) or 4 (2) transients per 512
increments, data were transformed using sinebell square
apodization in f1 and f2 to 4096 ꢁ 4096 points and spectral width
was 5656 ꢁ 5656 Hz (1) or 5952 ꢁ 5952 Hz (2); TOCSY spectra are
acquired using DIPSI spin lock and 150 ms missing time with 1 (1)
or 8 (2) transients per 1024 increments, data were transformed
using Gaussian apodization in f1 and f2 to 4096 ꢁ 4096 points
and spectral width was 5896 ꢁ 5896 Hz (1) or 5841ꢁ 5841 Hz (2);
ROESY acquisition with 300 ms mixing time and 16 (1) or 32 (2)
transients per 1024 (1) or 512 (2) increments, data transformation
with Gaussian apodization to 4096 ꢁ 4096 (1) or 8192 ꢁ 2048
(2) points and spectral width was 5981 ꢁ 5981 Hz (1) or
13298 ꢁ 13298 Hz (2); gHSQCAD with 2 (1) or 8 (2) transients per
1024 increments, data transformation with Gaussian apodization in
f1 and f2 to 4096 ꢁ 4096 points and f1 ꢁ f2 spectral width was
Esterified R1-barrigenol derivatives have already been charac-
terized as glycosylated compounds at C-3 from Pittosporum
tobira,[6] Pittosporum viridiflorum[7] and Pittosporum brevicalyx.[17]
Therefore, from a chemotaxonomic point of view, it was not
surprising to find a structural analog in another species of this
genus, P. senacia. Usually, the position at C-22 of R1-barrigenol
is substituted by an acyl moiety (tigloyl, angeloyl, 2-acetoxy-2-
methylbutanoyl, 3,3-dimethylacryloyl and 2-methylbutanoyl
units[6,7,12,14,15]). To the best of our knowledge, the glycosylation
at C-22 of R1-barrigenol is unusual.
O
30
29
O
21
22
12
26
25
Araf
OH
O
16
28
15
OH
GlcA
OH
OH
HO
27
3
HOOC
OH
O
O
O
HO
OH
O
O
HO
O
24
23
OH
O
Ara II
HO
O
OH
OH
Ara I
O
Glc
HO
HO
OH
Figure 3. Structure of compound 2.
Magn. Reson. Chem. (2012)
Copyright © 2012 John Wiley & Sons, Ltd.
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J. Linnek et al.
27137 ꢁ 5760 Hz (1) or 27137 ꢁ 5483 Hz (2); gHMBCAD with 8 (1) or
64 (2) transients per 1024 increments, data transformation with
Gaussian apodization in f1 and f2 to 4096 ꢁ 4096 points and f1 ꢁ f2
spectral width was 36199 ꢁ 5924 Hz (1) or 36199 ꢁ 5760 (2). Carbon
type (CH3, CH2 and CH): DEPT experiments. ESIꢀ (negative) and
HR-ESI-MS (positive): Q-TOF-1-Micromass spectrometer in m/z.
specimen (no. 26062010) was deposited in the Herbarium of the
Laboratory of Pharmacognosy, Burgundy University, Dijon, France.
Extraction and isolation
The dried and powdered branches of P. senacia (800g) were
refluxed with 70% MeOH for 1 h at 70–80ꢂC (2ꢁ 3 l). After filtration,
the solvent was removed by rotary evaporation yielding 30.12 g of
extract. An aliquot of this MeOH extract (8.00 g) was subjected to
VLC (silica gel, CHCl3/MeOH/H2O 65:32:6.5 and 100% MeOH) giving
19 main fractions: Fr.a-Fr.s. An aliquot of Fr.j (105.3mg) was
Plant material
The branches of P. senacia Putterlick were collected near Antsiranana
(Madagascar) in 2009 and identified by the botanist Natalie
Rafalimanana (University of Antsiranana, Madagascar). A voucher
Table 2. 13C NMR (150 MHz) and 1H NMR (600 MHz) data of sugar
moieties of compounds 1 and 2 in DMSO-d6 from 1D and 2D NMR
experiments (d in ppm, J in Hz)a
Table 1. 13C NMR (150 MHz) and 1H NMR (600 MHz) data of the
aglycones of compounds 1 and 2 in DMSO-d6 from 1D and 2D NMR
experiments (d in ppm, J in Hz)a
1
2
No.
dC
dH
dC
dH
1
2
GlcA
No.
dC
dH
dC
dH
1
103.6
78.5
77.9
75.1
77.7
171.6
4.28 d (6.9)
3.73 t (7.7)
3.68 m
103.6
78.5
77.7
74.8
77.0
171.8
4.28 d (6.9)
3.73 t (8.9)
3.68
1
38.1
25.4
88.4
38.5
55.0
17.6
32.2
38.9
47.0
36.4
22.8
121.6
143.3
41.2
27.1
22.4
47.7
41.2
0.88, 1.50
1.51, 1.78
2.99
38.3
25.6
88.3
38.5
54.7
17.8
35.5
40.5
46.1
36.2
23.0
124.0
143.2
46.4
66.1
71.8
47.8
41.8
0.89, 1.51
2
2
1.53, 1,81
3.02
3
3
4
3.55
3.57
4
—
—
5
3.38
3.42
5
0.70
0.72
6
6
1.27, 1.46
1.21, 1.38 m
—
1.44, ND
1.70 m, ND
—
Glc I
7
1
101.3
74.6
76.5
78.5
76.6
61.0
4.71 d (7.3)
3.00
101.4
74.6
76.3
71.8
76.5
61.2
4.72 d (7.3)
3.02
8
2
9
1.47
1.46
3
3.15
3.16
10
11
12
13
14
15
16
17
18
—
—
4
3.71
3.36
1.02 m, 1.79
5.16 br s
—
0.87, 1.81
5.32 br s
—
5
3.08 m
3.43, 3.65 m
3.08
6
3.45, 3.65
Glc II
—
—
1
94.0
72.3
76.7
69.5
77.7
60.6
5.24 d (8.2)
3.11
0.94, 1.71
1.58, 1.95
—
3.58
2
3.36
3
3.22 lt (8.5)
3.13
—
4
2.74 dd
(13.4, 3.6)
1.08, 1.62
—
2.41
5
3.14
6
3.44, 3.60
19
20
21
45.4
30.2
33.1
45.8
35.1
78.3
1.05, 2.41
—
Ara I
1
101.7
71.0
72.3
67.4
64.6
4.82 d (6.9)
3.43
101.9
70.5
72.6
67.6
65.0
4.81 d (6.0)
3.43
1.15 m,
1.33 m
1.49, 1.57
5.48 d (9.3)
2
3
3.35
3.32
22
31.5
4
3.62
3.62
22
78.5
27.2
16.1
15.4
16.8
20.1
61.7
29.1
19.6
4.03
5
3.37, 3.75 m
3.34, 3.76
23
27.5
16.0
15.1
16.6
25.4
175.1
32.6
23.2
0.97 s
0.75 s
0.86 s
0.69 s
1.08 s
—
0.98 s
0.76 s
0.89 s
0.91 s
1.27 s
3.12, 3.33
0.74 s
0.88 s
Ara II
24
1
104.6
70.9
72.4
67.6
65.4
4.14 d (5.7)
3.32
25
2
26
3
3.32
27
4
3.61
28
5
3.32, 3.63
29
0.88 s
0.87 s
Araf
1
30
107.1
80.2
76.9
84.8
61.7
5.06 br s
3.73
106.8
80.1
76.9
85.0
61.7
5.06 br s
3.78 m
at C-21
CH3CO
CH3CO
2
170.3
21.1
—
3
3.58
3.58
1.95 s
4
4.04 m
3.40, 3.48 m
4.05 m
5
3.42, 3.48 m
ND, not determined.
aOverlapped proton NMR signals are reported without designated
aOverlapped proton NMR signals are reported without designated
multiplicity.
multiplicity.
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Copyright © 2012 John Wiley & Sons, Ltd.
Magn. Reson. Chem. (2012)

Saponins from Pittosporum senacia
submitted to MPLC (RP-18 silica gel, H2O containing increasing
amounts of MeOH, 5 ml/min) to give eight subfractions, one of
them containing the pure compound 1, senaciapittoside A
(8.7 mg). In the same way, after another purification of an aliquot
of Fr.j (151.1 mg) by MPLC in the same conditions as previously
mentioned, 11 subfractions were obtained, one of them containing
the pure compound 2, senaciapittoside B (7.3mg).
References
[1] C. T. Sadashiva, P. Sharanappa, A. B. Remashree, A. V. Raghu, P. S.
Udayan, I. Balachandran. Advan. Biol. Res. 2010, 4, 301–304.
[2] N. J. Ferreira, I. G. Meireles de Sousa, Y. Cunha Luís, A. J. M. Currais, A.
C. Figueiredo, M. M. Cost, A. S. B. Lima, P. A. G. Sontos, J. G. Barroso, L.
G. Pedro, J. J. C. Scheffer. Flavour Fragr. J. 2007, 22, 1–9.
[3] C. Y. Ragasa, J. A. Rideout, D. S. Tierra, J. C. Coll. Phytochemistry 1997,
45, 545–547.
[4] A. Gurib-Fakim, F.-E. Demarne. Planta Med. 1994, 60, 584–585.
[5] M. F. Mahomoodally, A. Gurib-Fakim, A. H. Subratty. J. Food Sci. 2010,
75, M173–M177.
[6] I. D’Acquarica, M. C. Di Giovanni, F. Gasparrini, D. Misiti, C. D’Arrigo,
N. Fagnano, D. Guarnieri, G. Iacono, G. Bifulco, R. Riccio. Tetrahedron
2002, 58, 10127–10136.
Compound 1 (senaciapittoside A): white amorphous powder.
[a]2D5 –25.0 (c = 0.03, MeOH). For H NMR and 13C NMR data, see
1
Tables 1 and 2. HR-ESIMS m/z 1243.5729 [M + Na]+ (calculated
1243.5724). EISMS (negative mode) m/z 1219 [M–H]ꢀ and the
fragment 1087 [(M–H)–132]ꢀ.
[7] Y. Seo, J. M. Berger, J. Hoch, K. M. Neddermann, I. Bursuker, S. W.
Mamber, D. G. I. Kingston. J. Nat. Prod. 2002, 65, 65–68.
[8] R. Higuchi, T. Fujioka, M. Iwamoto, T. Komori, T. Kawasaki, E. V. Lassak.
Phytochemistry 1983, 22, 2565–2569.
[9] E. Mananjarasoa, M. Rakotovao, A. R. P. Ramanoelina, M. H.
Andriantsferana. J. Essent. Oil Res. 1998, 10, 459–462.
[10] A.-C. Mitaine-Offer, A. Marouf, B. Hanquet, N. Birlirakis, M.-A. Lacaille-
Dubois. Chem. Pharm. Bull. 2001, 49, 1492–1494.
[11] M. Kartal, A.-C. Mitaine-Offer, M. Abu-Asaker, T. Miyamoto, I. Calis, H.
Wagner, M.-A. Lacaille-Dubois. Chem. Pharm. Bull. 2005, 10,
1318–1320.
[12] G.-M. Fu, Y.-H. Wang, S. Gao, M.-J. Tang, S.-S. Yu. Planta Med. 2005,
71, 666–672.
[13] M. D. Greca, A. Fiorentino, P. Monaco, L. Previtera. Phytochemistry
1994, 35, 201–204.
[14] M. D. Greca, A. Fiorentino, P. Monaco, L. Previtera. Phytochemistry
1994, 36, 1479–1483.
[15] P. M. Gowri, S. V. S. Radhakrishnan, S. J. Basha, A. V. S. Sarma, J. M.
Rao. J. Nat. Prod. 2009, 72, 791–795.
[16] T. K. Tabopda, A.-C. Mitaine-Offer, T. Miyamoto, C. Tanaka, J.-F. Mirjolet,
O. Duchamp, B. T. Ngadjui, M.-A. Lacaille-Dubois. Phytochemistry 2012,
73, 142–147.
Compound 2 (senaciapittoside B): white amorphous powder.
1
[a]2D5 –9.5 (c = 0.05, MeOH). For H NMR and 13C NMR data, see
Tables 1 and 2. HR-ESIMS m/z 1305.5731 [M + Na]+ (calculated
1305.5728). EISMS (negative mode) m/z 1281 [M–H]ꢀ.
Acid hydrolysis and GC analysis
Each compound (3 mg) was hydrolyzed with 2 N aqueous TFA
(5 ml) for 3 h at 95 ꢂC. After extraction with CH2Cl2 (3 ꢁ 5 ml),
the aqueous layer was repeatedly evaporated to dryness with
MeOH until neutral and then analyzed by TLC over silica gel
(CHCl3/MeOH/H2O; 8:5:1) by comparison with authentic samples.
The trimethylsilyl thiazolidine derivatives of the sugar residue
of each compound were prepared and analyzed by GC.[18] The
absolute configuration was determined by comparing the
retention times with thiazolidine derivatives prepared in a similar
way from standard sugars (Sigma-Aldrich). L-Arabinose, D-glucose
and D-glucuronic acid were detected by coinjection of the deriva-
tives with standard silylated samples with tR 11.8, 18.6 and 15.2,
respectively, from compounds 1 and 2.
[17] C. Feng, B. G. Li, X. P. Gao, H. Y. Qi, G. L. Zhang. Arch. Pharm. Res.
2010, 33, 1927–1932.
[18] S. Hara, H. Okabe, K. Mihashi. Chem. Pharm. Bull. 1987, 35, 501–506.
Magn. Reson. Chem. (2012)
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