Other chemical constituents isolated from Solanum crinitum Lam. (Solanaceae)

Article abstract of DOI:10.1590/S0103-50532010001200007

The phytochemical investigation of Solanum crinitum Lam led to the isolation from the fruit trichomes of four flavonoids, tiliroside (1), astragalin (2), kaempferol (3), biochanin A-7-O-β-D-apiofuranosyl- (1→5)-β-D-apiofuranosyl-(1→6)-β-D-glucopyranoside (7), along with 4-hydroxybenzoic acid (12), and four cinnamic acid derivatives, cis-and trans-coumaric acids (10 and 11) and cis- and trans- ethyl coumarate (8 and 9). Three tri-glycosyl-steroidal alkaloids, solamargine (13), 20-epi-solamargine (14) and solasonine (16) were isolated from the methanolic extract of the green fruits. The derivatives 3,5,7,4′-tretra-O-methyl-kaempferol (4), 3,7,4′-tri- O-methyl-kaempferol (5), 3,7,4′-tri-O-methyl-5-O-acetyl- kaempferol (6), the peracetyl-episolamargine (15) and peracetyl-solasonine (17) were prepared. The structures were established through the analysis of their spectral data. The complete ¹H and ¹³C NMR data assignments of the new peracetyl derivatives of the alkaloids were made.

Full text of DOI:10.1590/S0103-50532010001200007

J. Braz. Chem. Soc., Vol. 21, No. 12, 2211-2219, 2010.
Printed in Brazil - ©2010 Sociedade Brasileira de Química
0103 - 5053 $6.00+0.00
Other Chemical Constituents Isolated from Solanum crinitum Lam. (Solanaceae)
Marli T. F. Cornelius,a Mário G. de Carvalho,*,a Tania M. S. da Silva,a
Cassia C. F. Alves,a Ana P. N. Siston,a Kelly Z. Alves,a Carlos M. R. Sant’Anna,a
Mario B. Neto,b Marcos N. Eberlinb andRaimundo Braz-Filhoc
aDepartamento de Química, ICE, Universidade Federal Rural do Janeiro, BR 465 km 07,
23890-000 Seropédica-RJ, Brazil
bInstituto de Química, Universidade Estadual de Campinas, 13084-862 Campinas-SP, Brazil
cSetor de Química de Produtos Naturais, LCQUI, CCT, Universidade Estadual do Norte Fluminense
Darcy Ribeiro, 28013-602 Campos dos Goytacazes-RJ, Brazil
O estudo fitoquímico de Solanum crinitum Lam forneceu quatro flavonóides: tilirosídeo (1),
astragalina (2), kaempferol (3) e biochanina A-7-O-b-D-apiofuranosil-(1→5)-b-D-apiofuranosil-
(1→6)-b-D-glucopiranosideo (7), ácido 4-hidroxibenzoico (12), e quatro derivados do ácido
cinâmico: cis- e trans- cumárico (10 e 11), cis- e trans-cumarato de etila (8 e 9), isolados
de tricomas do fruto. Do extrato metanólico de frutos verdes foram isolados três alcalóides
esteroidais glicosilados: solamargina (13), 20-epi-solamargina (14) e solasonina (16). Os derivados
3,5,7,4’-tetra-O-metil-kaempferol (4), 3,7,4’-tri-O-metil-kaempferol (5), 3,7,4’-tri-O-metil-5-O-
acetil-kaempferol (6), peracetil-epi-solamargina (15) e peracetil-solasonina (17) foram sintetizados
e estão sendo registrados pela primeira vez na literatura. As estruturas foram definidas através de
análise de dados espectrométricos.
The phytochemical investigation of Solanum crinitum Lam led to the isolation from the
fruit trichomes of four flavonoids, tiliroside (1), astragalin (2), kaempferol (3), biochanin A-7-
O-b-D-apiofuranosyl-(1→5)-b-D-apiofuranosyl-(1→6)-b-D-glucopyranoside (7), along with
4-hydroxybenzoic acid (12), and four cinnamic acid derivatives, cis- and trans-coumaric acids
(10 and 11) and cis- and trans- ethyl coumarate (8 and 9). Three tri-glycosyl-steroidal alkaloids,
solamargine (13), 20-epi-solamargine (14) and solasonine (16) were isolated from the methanolic
extract of the green fruits. The derivatives 3,5,7,4’-tretra-O-methyl-kaempferol (4), 3,7,4’-tri-
O-methyl-kaempferol (5), 3,7,4’-tri-O-methyl-5-O-acetyl-kaempferol (6), the peracetyl-epi-
solamargine (15) and peracetyl-solasonine (17) were prepared. The structures were established
through the analysis of their spectral data. The complete 1H and 13C NMR data assignments of the
new peracetyl derivatives of the alkaloids were made.
Keywords: Solanum crinitum, Solanaceae, steroidal glycoalkaloids, flavonoids, cinnamic acids
Introduction
against herbivores, against pathogenic microorganism2,3 and
molluscicidal activity,4 and are also of interest as starting
material for anabolic, anti-fertility, anti-inflammatory,
anti-allergic drugs.5 As the alkaloids, the flavonoids are
a frequent group of compounds in Solanum species, and
can be used as systematic markers for the family taxons.6-8
Solanum crinitum Lam (Solanaceae), popularly known
as “jurubeba” and “fruto-de-lobo”, is a shrub or small
tree in South America, including southern Brazil and
Colombia.9,10 In the course of our pharmacological and
phytochemical investigations of Solanum species,3,4,8,11-13
we have reported the cytotoxic and antitumoral activities
of flavonoids isolated from trichomes of young branches
Solanum (L) is the most representative genus of
Solanaceae, containing about 1,500 species, and 5,000
epithets. It is widespread in tropical and subtropical
regions of all the world, but its highest diversity occurs
in South America.1 Solanum species are a rich source of
steroidal alkaloids, flavonoids and their glycosides which
are known to possess a variety of biological activities.
The glycoalkaloids are natural toxins with ecological and
human health importance, such as the allelopathic effect
*e-mail: mgeraldo@ufrrj.br

2212
Other Chemical Constituents Isolated from Solanum crinitum Lam. (Solanaceae)
J. Braz. Chem. Soc.
and also of a rich glycoalkaloids total fraction from the
green fruits of Solanum crinitum Lam14 along with its
allelopathic activity.3
20-epi-solamargine (14) and solasonine (16) isolated
from the green fruits. The derivatives 3,5,7,4’-tetra-O-
methyl-kaempferol (4), 3,7,4’-tri-O-methyl-kaempferol
(5), 3,7,4’-tri-O-methyl-5-O-acetyl-kaempferol (6),
peracetyl-epi-solamargine (15) and peracetyl-solasonine
(17) were prepared and used to confirm the structures of
the natural compounds. The structures were established
through analysis of their spectral data, mainly NMR
(1D and 2D) and mass spectra. NMR data for the new
acetyl derivatives of the alkaloids are described herein
for the first time.
This work describes the isolation and structural
characterization of the flavonoids tiliroside (1),
astragalin (2), kaempferol (3) and biochanin A-7-O-b-
D-apiofuranosyl-(1→5)-b-D-apiofuranosyl-(1→6)-b-D-
glucopyranoside (7), cis- (10) and trans- (11) cumaric
acids and ethyl cis- (8) and trans- (9) cumarate and
4-hydroxybenzoic acid (12), which were isolated from
the trichomes, and the glycoalkaloids solamargine (13),

Vol. 21, No. 12, 2010
Cornelius et al.
2213
1
Results and Discussion
data.21-25 The integration of the H NMR signals allowed
us to calculate the approximate relative percentage of both
compounds: 36.16% of 8 and 63.84% of 9.
FromthetrichomesisolatedfromgreenfruitsofSolanum
crinitum Lam were isolated the flavonoids: tiliroside (1),
astragalin (2), kaempferol (3), and biochanin A-7-O-b-
D-apiofuranosyl-(1→5)-b-D-apiofuranosyl-(1→6)-b-D-
glucopyranoside (7), cis- (10) and trans- (11) cumaric
acids and ethyl cis- (8) and trans- (9) cumarate esters and
4-hydroxybenzoic acid (12); from green fruits methanolic
extracts were isolated three tri-glycosyl-steroidal alkaloids,
solamargine (13), 20-epi-solamargine (14) and solasonine
(16). The derivatives 3,5,7,4’-tetramethylkaempferol
(4), 3,7,4’-trimethylkaempferol (5), 3,7,4’-trimethyl-5-
acetylkaempferol (6), the peracetyl-20-epi-solamargine
(15) and peracetyl-solasonine (17) were prepared. The
structures were established on the basis of IR, NMR and
MS data analysis of the natural compounds and of the
derivatives 4-6, 15 and 17.
The fractions containing compounds 13, 14 and 16
showed a positive test for alkaloids. The detailed analysis
of the 1H and 13C NMR spectra of the isolated compounds
allowed to identify characteristic signals corresponding to
the same aglycone as that of the steroidal spirazolane-type
alkaloid in the three glycoalkaloids: four quaternary carbon
atoms, including one linked to oxygen and nitrogen atoms:
(dC 98.7 to 98.5 and one sp2 at dC 141.2 to 140.4), nine
methine groups (including two oxygenated atdC 78.5 to 78.0
and 81.1 to 78.6), ten methylene groups and four methyl
groups = (C)3(O-C-NH)(CH)7 (HC-O)2(CH2)10(CH3)4 =
C27H42NO3 = C27H42NO2 considering the presence of an
ether function (Table 1) having a trisaccharide moiety
(three anomeric carbon atoms: dC 100.6 to 100.3) attached
to the oxygen atom of carbon CH-3, with dC 78.5 to 78.0
(Table 1 and 2), which is significantly higher when
compared with the 13C chemical shift of the methyl carbon
CH-3 sustaining free hydroxyl group (about dC 71).
Compounds 1-3 were identified by the analysis of
13
1H and C NMR spectra, including HMQC and HMBC
experiments, and comparison with literature data for
tiliroside, kaempferol and astragalin.15-18 The first report
of 1 in Solanum species was made by Souza et al.14
Kaempferol and astragalin have been isolated from some
Solanum species.8 The treatment of 3 with diazomethane
yielded the methyl derivatives 4 and 5, described in the
literature,19 and the derivative 6 was obtained by treating
5 with Ac2O/Pyridine. The 2D NMR spectra, including
NOESY experiments, of these derivatives were used to
confirm the proposed structure of 3 and to carry out the
complete assignment of the 1H and 13C chemical shifts of
6 (see Experimental).
The 1H and 13C NMR spectra of compound 7 revealed
characteristic resonances of the isoflavonoid biochanin
A besides additional signals for three sugars unities, one
glucopyranoside and two apiofuranosides. Comparison
of these data with those of glycosides isolated from
Dalbergia nigra20 and Andira anthelmia,21 besides the
analysis of mass spectrum obtained by FAB-MS in
positive mode {m/z 733.20820 ([M+Na]+, C32H38O18+Na+,
calc.: m/z 733.19558), m/z 601.12300 [M+Na-132]+, m/z
451.0600 (M+Na-282)+, m/z 449.27710 [M+Na-284]+, m/z
431.16210 [M+Na-302]+, m/z 317.1338 [M+Na-416]+, m/z
287.0865 [M+Na-446]+}, led us to identify the compound
as biochanin A, 7-O-b-D-apiofuranosyl-(1→5)-b-D-
apiofuranosyl-(1→6)-b-D-glucopyranoside (7). This is
the first report of this apiofuranosyl derivative in Solanum
species.
The H and 13C chemical shifts of the trisaccharide
1
moieties of 13 and 14, had practically the same values
(Table 2), indicating identical the partial structure O-[a-
L-rhamnopyranosyl-(1→2)-O-[a-L-rhamnopyranosyl-
(1→4)]-b-D-glucopyranoside = C18H32O13 (three degrees
of unsaturation, C18H38O13 – C18H32O13 = H6). On the other
hand, the 1H and 13C signals observed in NMR spectra of
16 allowed to characterize the trisaccharide as O-[a-L-
rhamnopyranosyl-(1→2)-O-[b-glucopyranosyl-(1→3)]-
b-D-galactopyranoside = C18H32O14 (three degrees of
unsaturation, C18H38O14 – C18H32O14 = H6). All these NMR
spectral data allowed to deduce the same molecular formulas
C45H73NO15 to 13 and 14 and C45H73NO16 to 16, all with
ten degrees of unsaturation corresponding to one double
bond and six rings). In fact, the values of pseudomolecular
peaks in the positive HRMS spectra at m/z 868.5239
(M + H•+, 100%) of 13 (C45H74NO15 = m/z 868.5058) and
at m/z 868.5305 (M + H•+, 100%) of 14 (C45H74NO15 = m/z
868.5058), besides additional peaks compatible with loss
of sugar moieties, were used to confirm the molecular
formula of these isomeric compounds (13 and 14). The
1
hydrogen and carbon atoms signals observed in the H
and 13C NMR spectra of 13-17 (Tables 1 and 3) were also
assigned with aid of the homonuclear 2D 1H-1H-COSY and
heteronuclear 2D HMQC (1H-13C-COSY-1JCH) and HMBC
(1H-13C-COSY-nJCH, n = 2 and 3), allowing to identify the 1H
chemical shifts of the methyl group signals of the aglycone
and of the trisaccharide moieties (Table 3 and 4): dH 0.83
(d, 3H-27), 0.88 (s, 3H-18), 1.07 (s, 3H-19) and 1.09 (d,
3H-21) of 13, 0.79 (d, 3H-27), 0.85 (s, 3H-18), 1.06 (s,
1
Compounds 8-12 were identified by the H and
13C NMR spectral data analysis of the mixtures of 8+9,
10+11 and of 12 along with comparison with literature

2214
Other Chemical Constituents Isolated from Solanum crinitum Lam. (Solanaceae)
J. Braz. Chem. Soc.
3H-19) and 1.28 (d, 3H-21) of 14, and 0.76 (d, 3H-27), 0.82
(s, 3H-18), 1.03 (s, 3H-19) and 1.17 (d, 3H-21) of 16; signals
of the hydrogen H-6 linked to sp2 carbons of double bounds
at dH: 5.32 (13), 5.33 (14), 5.30 (16), besides the signals of
H-16 located in the spyrazolane ring (d, J 6 Hz) at dH 4.42
(13); 4.82 (14) and 4.21 (16). The additional doublets
(J 6 Hz) corresponding to methyl groups were revealed at
d3H 1.64 and d3H 1.78 in the spectra of 13 and 14 suggested
the presence of two rhamnose moieties in both compounds
and only one rhamnose for 16 as indicated by signal at
dH 1.78. The differences observed in the 1H NMR spectra
of 13 and 14, recorded in same apparatus (1H: 500 MHz;
13C: 125 MHz) and solvent pyridine-d5, involving mainly
the chemical shifts of the hydrogen atoms H-16 (dH 4.42
and 4.82), H-17 (dH 1.78 and 2.02), H-20 (dH 1.98 and 2.14),
2H-26 (dH 2.78 and 3.04/2.89) and 3H-21 (dH 1.09 and
1.28), which revealed correlation in the HMQC with the 13C
signals of the corresponding carbon atoms: CH-16 (dC 79.2
and 81.1), CH-17 (dC 63.9 and 63.0), CH-20 (dC 42.0 and
42.0), CH2-26 (dC 48.2 and 47.1) and 3H-21 (dC 16.1 and
15.7), as summarized in Figure 1. Comparative analysis
of these data was used to suggest the two stereoisomers
H-20b (13) and H-20a (14), since the 22aN and 22bN
possibilities were eliminated considering the absence of
13C signal corresponding to a methylene carbon CH2-23 at
about dC 27 which is consistent with the 13C NMR chemical
shift of the 22bN stereoisomer (Figure 1). The b-position
of the methyl Me-21 located at carbon atom 20 of 14 may
be used to justify the 13C chemical shifts of the CH-16
(dC 81.1, absence of the g-effect from Me-21) and CH2-23
(dC 34.0, presence of the g-effect from Me-21).26-30 In order
to compare the relative stabilities of the epimeric structures
13/14, a molecular modeling study was implemented using
the Spartan 06 for Linux program (Wavefunction, Inc.), see
Supplementary Information.31
The complete H and C chemical shift assignments
of the signals of CH3, CH2, CH and C observed in the 1H
(including 1H-1H-COSY) and 13C ({1H} and DEPT) NMR
spectra (including 2D experiments HMQC and HMBC)
(Tables 1, 2, 3 and 4) and comparison with values described
in the literature for solamargine (13)26-29 and solasonine
(16)28,31 led to the proposition of the structures (25R)
(20S)(16S)-3b-{O-a-L-rhamnopyranosyl-(1→2)-O-[a-L-
rhamnopyranosyl-(1→4)]-b-D-glucopyranosyl}-22aN-
spirosol-5-en (solamargine, 13), the new glycoalkaloid
(25R)(20R)(16S)-3b-{O-a-L-rhamnopyranosyl-(1→2)-
O-[a-L-rhamnopyranosyl-(1→4)]-b-D-glucopyranosyl}-
22aN-spirosol-5-en (16-epi-solamargine, 14), and (25R)
(20S)(16S)-3b-{O-a-L-rhamnopyranosyl-(1→2)-O-[b-
glucopyranosyl-(1→3)]-b-D-galactopyranosyl} -22aN-
spirosol-5-en (solasonine, 16) for the three steroidal
glycoalkaloids isolated from this plant (Figure 1).
1
13
Figure 1. Comparative analysis of the chemical shifts corresponding to the hydrogen (dH) and carbon (dC) atoms CH-16, CH-17, CH-20, CH3-21, C-22,
CH2-23 and CH2-26 of 13 and 14, including comparison of the dC these carbon atoms and of the CH2-24 to reveal the structural distinction between
solamargine and b-solamarine.

Vol. 21, No. 12, 2010
Cornelius et al.
2215
Table 1. 13C NMR spectral data for aglycone of 13, 14 and 16 in pyridine-d5
(125 MHz: 13 and 14; 50 MHz: 16) and of peracetyl derivatives 15 and
17 in CDCl3 (125 MHz)*
Experimental
General procedure
C
13
37.9
30.5
78.5
39.3
141.2
122.2
32.1
32.0
50.7
37.5
21.5
40.5
41.0
57.0
32.9
79.2
63.9
16.9
19.8
42.0
16.1
98.7
35.0
31.4
32.1
48.2
20.1
14
37.5
30.2
78.0
39.0
140.4
121.9
32.3
31.7
50.3
37.2
21.1
39.7
40.8
56.5
32.6
81.1
63.0
16.4
19.4
42.0
15.7
98.5
34.0
30.0
31.7
47.1
19.2
15a
37.3
29.7
79.3
38.4
140.4
122.2
31.8
31.6
50.3
36.9
21.0
40.0
42.9
53.0
31.6
75.9
62.1
12.8
19.4
40.0
21.1
99.7
35.2
30.1
29.1
45.5
18.5
16
37.6
30.2
78.4
38.8
141.0
121.8
32.3
31.8
50.3
37.2
21.2
39.9
40.8
56.6
32.6
78.6
63.2
16.5
19.5
41.9
15.8
98.5
34.3
30.5
30.8
47.5
19.5
17a
37.1
29.5
79.8
38.4
140.2
121.9
31.6
31.4
50.0
36.7
20.6
39.8
42.7
54.8
31.6
75.6
62.0
12.4
20.3
39.7
21.4
99.2
35.0
30.8
28.9
45.2
19.2
Melting points have not been corrected. IR, NMR and
mass spectra were recorded on the same equipments used
in previous papers.21,32 Column chromatography was carried
out with silica gel (Vetec and Aldrich 0.05-0.20 mm) and
Sephadex LH-20 (Sigma, USA); silica gel F254 G (Vetec)
was used for preparative TLC; aluminum backed (Sorbent)
silica gel plates W/UV254 were used for analytical TLC,
with visualization under UV (254 and 366 nm), with AlCl3-
ETOH (1%), Liebermann-Burchard and/or Godin reagents,
or exposure to iodine vapor.
1 (CH2)
2 (CH2)
3 (CH)
4 (CH2)
5 (C)
6 (CH)
7 (CH2)
8 (CH)
9 (CH)
Plant material
10 (C)
11 (CH2)
12 (CH2)
13 (C)
The green fruits of Solanum crinitum Lam. were
collected in September, 2001, in the campus of Universidade
Federal Rural do Rio de Janeiro (UFRRJ), Seropédica-RJ,
Brazil. They were collected by M.Sc. José Milton Alves
(Agronomy Institute, UFRRJ). The identification was made
by Dr. Maria de Fátima Agra, Laboratório de Tecnologia
Farmacêutica, Universidade Federal da Paraíba, João
Pessoa-PB. A voucher specimen (No. JP-28000) was
deposited at the Herbarium Prof. Lauro Pires Xavier,
Universidade Federal da Paraíba, João Pessoa-PB, Brazil.
14 (CH)
15 (CH2)
16 (CH)
17 (CH)
18 (CH3)
19 (CH3)
20 (CH)
21 (CH3)
22 (C)
Extraction and isolation
The trichomes (9.7 g) were isolated by scraping the
green fruits with a glass slide and were subsequently
extracted with CHCl3 and MeOH in an ultrasound bath to
furnish the CHCl3 extract (310.0 mg) and MeOH (3.0 g)
residues. The CHCl3 residue was chromatographed on a
sephadex CC, using MeOH as eluent, and 13 fractions were
collected and analyzed by TLC and 1H NMR spectroscopy.
Fractions 7-8 yielded astragaline (1, 10.0 mg) and fractions
9-10 yielded the tilirozide (2, 25.0 mg). The methanolic
extract was chromatographed on a silica gel column (col
A) and 20 fractions were collected and analyzed by TLC
plate. Fractions 5-8 (2.0 g) were dissolved in methanol and
addition of CH2Cl2 yielded a precipitate that was separated
by filtration to afford a solid (1.6 g) that was identified
23 (CH2)
24 (CH2)
25 (CH)
26 (CH2)
27 (CH3)
*Number of hydrogens bound to carbon atoms deduced by comparative
analysis of {1H}- and DEPT-13C NMR spectra. 2D H-1H-COSY and
1
1H-13C-nJCH (n = 2 and 3) NMR spectra were also used in these assignments;
a
d
COCH : 170.1-170.2 (CO), 23.8 (CH ).
3
3
The natural alkaloids 14 and 16 were treated with
Ac2O/pyridine to yield the peracetyl derivatives 15 and
17. The 1H and 13C NMR spectral data of these peracetyl
derivatives (15 and 17), obtained through the analysis of
extensive 1D and 2D NMR experiments (Tables 1-4), were
also used to confirm the postulated structures to 14 and 16.
The analysis of 1D and 2D NMR spectra was also used to
make the complete hydrogen and carbon-13 chemical shift
assignments for the alkaloid, the new 16-epi-solamargine
(14) and for the two peracetyl derivatives 15 and 17.
o
as kaempferol (3, mp 282-284 C) and the mother liquor
(AM-5-8). The reaction of kaempferol (65.0 mg) with
diazomethane yielded 16.1 mg of 4 (mp 128-130 oC) and
36.6 mg of 5 (mp 140-142 °C) that were separated by
preparative TLC. 20.3 mg of 5 were treated with Ac2O/
pyridine (1:1) to afford 6 (14.8 mg, mp 166-168 °C). The
mother liquor (AM 5-8, 360.0 mg) was submitted to silica

2216
Other Chemical Constituents Isolated from Solanum crinitum Lam. (Solanaceae)
J. Braz. Chem. Soc.
Table 2. 13C NMR spectral data for glycoside moieties of 13, 14 and 16 in pyridine-d5 (125 MHz: 13 and 14; 50 MHz: 16) and of peracetyl derivatives
15 and 17 in CDCl3 (125 MHz)*
C
trisaccharide of 13
100.6
78.2
trisaccharide of 14
100.3
77.8
trisaccharide of 15a
99.8
trisaccharide of 16
100.4
76.6
trisaccharide of 17a
99.6
1’ (CH)
2’ (CH)
3’ (CH)
4’ (CH)
5’ (CH)
6’ (CH2)
1” (CH)
2” (CH)
3” (CH)
4” (CH)
5” (CH)
6” (CH3)
1’’’ (CH)
2’’’ (CH)
3’’’ (CH)
4’’’ (CH)
5’’’ (CH)
6’’’ (CH3)
76.5
73.1
78.3
78.1
75.5
84.9
78.3
79.0
78.8
77.9
70.5
69.5
77.3
76.9
72.5
77.5
70.9
61.7
61.3
62.1
62.6
62.0
103.3
72.9
102.9
72.5
99.7
105.9
72.6
99.2
70.5
71.8
73.1
72.8
68.8
80.2
72.3
74.3
73.9
70.8
75.2
68.2
70.8
70.5
68.1
77.5
72.0
18.9
18.5
17.4
62.6
60.9
102.4
72.9
102.1
72.6
97.5
102.3
72.9
97.0
70.2
68.9
73.1
72.9
68.8
74.2
69.1
74.5
74.2
71.5
75.0
70.9
69.9
69.5
66.7
69.5
66.5
19.0
18.7
17.5
18.7
17.1
*Number of hydrogens bound to carbon atoms deduced by comparative analysis of {1H}- and DEPT-13C NMR spectra. 2D 1H-1H-COSY and 1H-13C-nJCH
(n = 2 and 3) NMR spectra were also used in these assignments; a dCOCH : 169.9-170.9 (CO), 20.9-21.7 (CH ).
3
3
gel CC and fractions 8-12 yielded a mixture of 8 and 9
(1.2 mg, gum). Fraction 10 (850 mg) from col A was
chromatographed on a silica gel column and 130 fractions
of 50 mL were collected. Fractions 32-40 yielded 35.5 mg
of a material corresponding to the mixture of 10, 11 and 12.
Fractions 94-114 (102.3 mg) were chromatographed on a
silica gel column and fractions 3-5 yielded 7 (9.5 mg) after
filtration on Sephadex LH20.
The green fruits (2.6 g) of Solanum crinitum Lam were
powdered and extracted with ethanol + acetic acid (2%) and
900 mL of solution was obtained. 900 mL of acetic acid
(10%) were added, and the solution was left to stand and
chill overnight.The solution was filtered under vacuum using
Hirsch funnel. NH4OH (pH 9-10) was added to the filtrate
which was allowed to stand overnight in the fridge affording
a precipitate (96.4 g) corresponding to the glycoalkaloids
fraction was obtained. 90 g of the glycoalkaloids was
adsorbed on silica gel and applied on a silica gel column,
eluted with hexane, dichloromethane, ethyl acetate, acetone
andmethanol.14fractionsof500mLwerecollected.Fraction
8 (1.0 g), collected with ethyl acetate, was chromatographed
in a silica gel CC and fractions analyzed by silica gel TLC
plate. Fraction 84 (52.6 mg) from this column was filtered
on a Sephadex LH20 column, eluted with methanol, and the
alkaloid solamargine 13 (268-270 °C) was obtained. Fraction
10 (8.67 g) was extracted with methanol and the alkaloids
were detected with Dragendorff and Libermann Burchard.
This fraction was chromatographed on a silica gel column
using CH2Cl2/MeOH (3:1) as initial eluent. 79 fractions were
collected and were analyzed by TLC plate and reunited in
group. Fractions 24-29 (1.2 g) were filtered on a Sephadex
LH-20 column and the epi-solamargine (14, 225.8 mg,
238-250 °C) was obtained. Acetic anhydride:pyridine (1:1)
treatment of 14 (44.5 mg) yielded 44.5 mg of the peracetyl
derivative 15 (mp 128-130 °C). Fractions 39-44 (1.3 g)
were filtered on Sephadex LH20 to isolate solasonine (16,
93.5 mg, mp 244-246 °C). Treatment of 16 (79.1 mg) with
aceticanhydride:pyridine(1:1) gavethe peracetyl-solasonine
(17, 27.6 mg, mp 148-150 °C).
3,7,4’-trimethoxy-5-acethoxyflavone (6)
Yellow powder, mp 166-168 °C; IR (KBr) nmax/cm-1:
1
1762, 1632, 1606, 1511; H NMR (CDCl3, 500 MHz) d
6.60 (d, J 2.5 Hz), 6.82 (d, J 2.5 Hz), 8.02 (dd, J 9.0 Hz),
7.02 (d, J 9.0 Hz), 3.78, 3.90 (s), 3.89 (s), 2.47 (s), 13C NMR
(CDCl3, 125 MHz) d154.8 (C-2), 140.8 (C-3), 173.2 (C-4),
150.5 (C-5), 108.0 (CH-6), 163.1 (C-7), 98.6 (CH-8), 157.8
(C-9), 111.4 (C-10), 122.9 (C-1’), 129.9 (CH-2’,6’), 114.0
(CH-3’,5’), 161.4 (C-4’), 59.9, 55.9, 55.3 (MeO-3,7,4’,
respectively), 169.7/21.2 (COCH3-5).

Vol. 21, No. 12, 2010
Cornelius et al.
2217
Table 3. 1H NMR spectral data for aglycone of 13, 14 and 16 in pyridine-d5 (500 MHz: 13 and 14; 200 MHz: 16) and of peracetyl derivatives 15 and 17
in CDCl3 (500 MHz). Chemical shifts in d (ppm) and coupling constants (J in parentheses) in Hz*
H
1
2
3
4
13
14
15a
16
17a
1.73/1.00
2.10/1.87
3.89 (m)
2.82/2.75
1.75/1.02
2.08/1.85
3.89 (m)
1.85/1.08
1.93/1.58
3.60
1.85/1.08
1.95/1.25
3.58 (m)
2.45/2.28
3.93 (m)
2.87/2.76
2.81 (dd, J 11.6; 3.1)
2.73 (t, J 11.6 )
2.40/2.25
6
5.32 (br s)
1.90
5.33 (br s)
1.85
5.36 (m)
1.94
5.30 (br s)
5.38 (m)
2.30/1.68
1.53
7
8
1.48
1.58
1.56
9
0.90
0.89(m)
1.42
0.95
0.95
11
12
14
15
16
17
18
19
20
21
23
24
25
26
27
1.45
1.55-1.45
2.08/1.25
0.98
1.55-1.45
2.08/1.25
0.98
1.63/1.05
1.10
1.68/1.09
1.09 (m)
2.08/1.48
4.82
2.08/1.45
4.42
2.20/1.70
5.06
1.95/1.85
5.08
4.21
1.78
2.02
1.53
1.53
0.88 (s)
1.07 (s)
1.98
0.85 (s)
0.87 (s)
1.01 (s)
2.10
0.82 (s)
1.03 (s)
0.91 (s)
1.04 (s)
2.08
1.06 (s)
2.14
1.09 (d, J 6.9)
1.90/1.65
1.52
1.28 (d, J 6.7)
1.92/1.78
1.65
1.26 (d, J 6.9)
2.36/1.04
2.28/1.68
2.02
1.17 (d)
1.26 (d, J 7.0)
2.35/1.03
1.70/1.65
2.02
1.65
1.58
2.78
3.04 (m)/2.89 (t, J 11.5)
3.20
2.83
3.16(m)
0.83 (d, J 4.8)
0.79 (d, J 6.5)
0.92 (d, J 6.6)
0.76(d)
0.93 (d, J 7.5)
*Number of hydrogens bound to carbon atoms deduced by comparative analysis of {1H}- and DEPT-13C NMR spectra. Chemical shifts and coupling
constants (J, in parentheses) obtained from 1D 1H NMR spectra. 2D 1H-1H-COSY and 1H-13C-nJCH (n = 2 and 3) NMR spectra were also used in these
assignments; a dCOCH : 2.16 (s).
3
Epi-solamargine (14)
C63H92NO24: 1246.600928), m/z 1204.5278 (calc. for
C61H90NO23: 1204.590364) [M+H-42]+, m/z 974.5256
(calc. for C51H76NO17: 974.511325) [M+H-rhamnose]+, m/z
932.5163 (calc. for C49H74NO16: 932.500760) [M-314]+, m/z
634.7963 (calc. for C39H56NO6: 634.410763) [M+H-612]+,
m/z 413.2715 (calc. for C27H43NO2: 413.329379)
[M-acetate-rhamnose-rhamnose-glucose]+. 13C RMN:
Tables 1 and 2. 1H RMN: Tables 3 and 4.
Colorless crystals, mp 238-250 °C; [a]25D – 98 (c 0.1,
MeOH); IR (KBr) nmax/cm-1: 3417, 2927, 1453, 1384,
1044 cm-1. HR-FABMS positive ions at m/z 868.5239
[M+H]+ (calc. for C45H74NO15: m/z 868.505846), 722.4643
(calc. for C39H64NO11: 722.447937) [M+H-rhamnose]+,
m/z 576.4037 (calc. for C33H54NO7: 576.390028) [M+H-
rhamnose-rhamnose]+, m/z 445.7553 (calc. for C21H33O10:
445.207372) [M-423]+, m/z 414.3467 (calc. for C27H44NO2:
414.337204) [M+H-rhamnose-rhamnose-glucose]+, m/z
413.2782 (calc. for C27H43NO2: 413.329379) [M-rhamnose-
rhamnose-glucose]+. 13C RMN: Tables 1 and 2. 1H RMN:
Tables 3 and 4.
Peracetyl-solasonine (17)
Crystal, mp 148-150 °C; 13C RMN: Tables 1 and 2;
1H RMN: Tables 3 and 4.
Supplementary Information
Peracetyl-epi-solamargine (15)
Crystal, mp 128-130 °C, [a]25D – 165 (c 0.1, MeOH);
IR (KBr) nmax/cm-1: 2943. 2728, 1751, 1438, 1374, 1045;
Positive ion FABMS m/z 1246.5522 [M+H]+ (calc. for
Supplementary data associated with this paper are
available free of charge at http://jbcs.sbq.org.br, as a PDF
file including a molecular modeling study in order to

2218
Other Chemical Constituents Isolated from Solanum crinitum Lam. (Solanaceae)
J. Braz. Chem. Soc.
Table 4. 1H NMR spectral data for glycoside moieties of 13, 14 and 16 in pyridine-d5 (500 MHz: 13 and 14; 200 MHz: 16) and of peracetyl derivatives
15 and 17 in CDCl3 (500 MHz). Chemical shifts in d (ppm) and coupling constants (J in parentheses) in Hz*
H
Glycoside unit of 13
4.95
Glycoside unit of 14
Glycoside unit of 15a
4.56 (d, J 7.7)
3.56
Glycoside unit of 16
Glycoside unit of 17a
4.46 (d, J 8.0)
1’
2’
3’
4’
5’
6’
4.96
4.22
4.95
3.98
4.24
3.84 (dd, J 8.0; 9.5)
3.91 (dd, J 9.5; 3.0)
5.26 (br s)
4.23 (t)
4.23
5.28 (t, J 9.4)
3.73 (t, J 9.4)
3.59 (m)
4.33 (m)
4.82
4.42 (t)
4.40
3.65 (br d, J 9.2)
3.66 (td, J 9.2)
3.79 (m)
4.22(br d, J 12.0)
4.10 (br d, J 12.0)
4.23 (dd, J 12.1; 3.2)n
4.10 (dd, J 12.1; 3.2)
4.44 (d, J 12.3)
4.28 (dd, J 12.3; 3.6)
4.12 (m)
4.06 (m)
1”
5.86 (s)
4.69 (sl)
5.87 (s)
4.69(d, J 1.4)
4.55 (dd, J 9.2; 3.2)
4.34 (t, J 9.2)
4.93 (m)
4.79 (d, J 1.5)
5.02 (m)
5.17(d)
4.71 (d, J 8.0)
4.85 (dd)
5.22 (t)
2”
4.68 (sl)
3”
4.55 (dd, J 9.1; 2.9)
4.38
5.16 (dd, J 10.2; 3.1)
5.03 (t, J 10.2)
3.85 (m)
4”
5.08
5”
4.92
3.69 (m)
6”
1.64 (d, J 5.9)
6.41 (s)
1.64 (d, J 6.1)
6.41 (s)
1.14 (d, J 6.1)
4.90 (d, J 1.5)
5.00 (m)
4.38 (br d, J 10.5)/4.10
5.02 (sl)
1’’’
2’’’
3’’’
4’’’
5’’’
6’’’
6.28 (s)
4.90 (d)
4.61
4.84 (br s)
4.64 (dd, J 9.1; 2.9
4.34
4.84 (d, J 1.8)
4.64 (dd, J 9.2; 3.3)
4.32 (t, J 9.2)
4.98 (m)
5.22
5.23 (dd, J 10.1 Hz; 3.1)
5.05 (t, J 10.1)
4.36 (m)
5.22
5.02(m)
4.96
4.88 (br s)
1.78 (d)
4.43 (m)
1.78 (d, J 6.0)
1.78 (d, J 6.2)
1.17 (d, J 6.2)
1.19 (d, J 6.0 )
*Number of hydrogen bound to carbon atoms deduced by comparative analysis of {1H}- and DEPT-13C NMR spectra. Chemical shifts and coupling
constants (J, in parentheses) obtained from 1D 1H NMR spectra. 2D 1H-1H-COSY and 1H-13C-nJCH (n = 2 and 3) NMR spectra were also used in these
assignments; a dCOCH : 1.98-2.15 (8×CH ).
3
3
compare the relative stabilities of the epimeric structures
13 and 14.
5. Mola, J. L.;Araujo, E. R.; Magalhães, G. C.; Quim. Nova 1997,
20, 460.
6. D’arcy, W. G. In The Biology and Taxonomy of the Solanaceae;
Hawkes, J. G.; Lester, R. N.; Skelding,A., eds.,Academic Press:
London, 1979, pp. 3-49.
Acknowledgments
The authors are grateful to CNPq, FAPERJ and
CAPES for research fellowships and financial support.
They also thank CENAUREMN for the 500 MHz NMR
spectra, Laboratório Plataforma de Métodos Analíticos de
Farmanguinhos/FIOCRUZ for the 400 MHz NMR spectra.
They thank MSc J.M. Alves and Dr. M. de F. Agra for the
collection and classification of plant material.
7. Steinharter, T. P.; Cooper-Driver, G. A.; Anderson, G. J.;
Biochem. Syst. Ecol. 1986, 14, 299.
8. Silva, T. M. S.; Carvalho, M. G. de; Braz-Filho, R.; Agra, M.
F.; Quim. Nova 2003, 26, 517.
9. Agra,M.F.;PhDThesis,UniversidadedeSãoPaulo,Brazil,2000.
10. Dias-Filho, M. B.; Pesq. Agropec. Bras. 1997, 32, 789.
11. Silva, T. M. S.; Nascimento, R. J. B.; Câmara, C. A.; Castro, R.
N.; Braz-Filho, R.;Agra, M. de F.; Carvalho, M. G. de; Biochem.
Syst. Ecol. 2004, 32, 513.
References
12. Silva, T. M. S. da; Braz-Filho, R.; Carvalho, M. G. de; Agra,
M. de F.; Biochem. Syst. Ecol. 2002, 30, 1083.
1. Agra, M. F.; Novon 1999, 9, 292.
2. Fukuhara, K.; Shimizu, K.; Kubo, I.; Phytochemistry 2004, 65,
1283.
13. Silva, T. M. S. da; Silva, C. de C.; Braz-Filho, R.; Carvalho, M.
G. de; Agra, M. de F.; Rev. Bras. Farmacogn. 2002, 12, 85.
14. Souza, A. E.; Silva, T. M. S. da; Alves, C. C. F.; Carvalho, M.
G. de; Braz-Filho, R.; Echevarria,A.; J. Braz. Chem. Soc. 2002,
13, 838.
3. Alves, C. C. F.; Alves, J. M.; Silva, T. M. S.; Carvalho, M. G.
de; Neto, J. J.; Floresta e Ambiente 2003, 10, 93.
4. Silva, T. M. S.; Câmara, C. A.; Agra, M. F.; Carvalho, M. G.
de; Frana, M. T.; Brandoline, S. V. P. B.; Paschoal, L. S.; Braz-
Filho, R.; Fitoterapia 2006, 77, 449.
15. Markham, K. R. In Methods in Plant Biochemistry; Harborne,
J. B.; Mabry, T. J., eds.,Academic Press: London, 1989, p. 197.

Vol. 21, No. 12, 2010
Cornelius et al.
2219
16. Kaouadji, M.; Phytochemistry 1990, 29, 2295.
25. Scott, K. N.; J. Am. Chem. Soc. 1972, 94, 8564.
26. Ripperger, H.; Phytochemistry 1995, 39, 1475.
17. Agrawal, P. K.; Bansal, M. C. In Carbon-13 NMR of Flavonoids,
StudiesinOrganicChemistry;Elsevier:Amsterdam,1989,p.362.
18. Markham, K. R.; Geiger, H. In The Flavonoids Advances in
Research Since 1986; Harborne, J. B., ed., Chapman & Hall:
London, 1994, pp. 441.
27. Lorey, S.; Porzel, A.; Ripperger, H.; Phytochemistry 1996, 41,
1633.
28. Wanyonyi, A. W.; Chiabra, S. C.; Mkoji, G.; Eilert, U.; Njue,
W. M.; Phytochemistry 2002, 59, 79.
19. Dong, H.Y.; Gou, L.; Cao, S. G.; Chen, S. X.; Sim, K.Y.; Goh,
S. H.; Kini, R. M.; Phytochemistry 1999, 50, 899.
20. Mathias, L.; Vieira, I. J. C.; Braz-Filho, R.; Rodrigues-Filho,
E. A.; J. Nat. Prod. 1998, 61, 1158.
29. Weissenberg, M.; Phytochemistry 2001, 58, 501.
30. Usubillaga, A.; Aziz, I.; Tettamanzi, M. C.; Waibel, R.;
Phytochemistry 1997, 44, 537.
31. Lee, C.; Yang, W.; Parr, R. G.; Phys. Rev. B: Condens. Matter
Mater. Phys. 1988, 37, 785.
21. Silva, V. C. da; Carvalho, M. G. de; Silva, S. L. da C. e; Rev.
Latinoam. Quim. 2007, 35, 13.
32. Carvalho, M. G. de; Silva, V. C. da; Silva, T. M. S. da; Câmara,
C. A.; Braz-Filho, R.; An. Acad. Bras. Cienc. 2009, 81, 21.
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Submitted: March 23, 2009
24. Silva, A. M. S.; Alkorta, I.; Elguero, J.; Silva, V. L. M.; J. Mol.
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Published online: August 12, 2010
FAPESP has sponsored the publication of this article.

J. Braz. Chem. Soc., Vol. 21, No. 12, S1-S9, 2010.
Printed in Brazil - ©2010 Sociedade Brasileira de Química
0103 - 5053 $6.00+0.00
Other Chemical Constituents Isolated from Solanum crinitum Lam. (Solanaceae)
Marli T. F. Cornelius,a Mário G. de Carvalho,*,a Tania M. S. da Silva,a
Cassia C. F. Alves,a Ana P. N. Siston,a Kelly Z. Alves,a Carlos M. R. Sant’Anna,a
Mario B. Neto,b Marcos N. Eberlinb andRaimundo Braz-Filhoc
aDepartamento de Química, ICE, Universidade Federal Rural do Janeiro, BR 465 km 07,
23890-000 Seropédica-RJ, Brazil
bInstituto de Química, Universidade Estadual de Campinas, 13084-862 Campinas-SP, Brazil
cSetor de Química de Produtos Naturais, LCQUI, CCT, Universidade Estadual do Norte Fluminense
Darcy Ribeiro, 28013-602 Campos dos Goytacazes-RJ, Brazil
Figure S1. 1H NMR spectrum of compound 1.
*e-mail: mgeraldo@ufrrj.br

S2
Other Chemical Constituents Isolated from Solanum crinitum Lam. (Solanaceae)
J. Braz. Chem. Soc.
Figure S2. 1H NMR spectrum of compound 2.
Figure S3. 1H NMR spectrum of compound 3.

Vol. 21, No. 12, 2010
Cornelius et al.
S3
Figure S4. 13C NMR spectrum of compound 4.
Figure S5. 1H NMR spectrum of compound 5.

S4
Other Chemical Constituents Isolated from Solanum crinitum Lam. (Solanaceae)
J. Braz. Chem. Soc.
Figure S6. 1H NMR spectrum of compound 6.
Figure S7. 1H NMR (500 MHz) of compound 7.

Vol. 21, No. 12, 2010
Cornelius et al.
S5
Figure S8. 1H NMR spectrum(500 MHz) of compounds 8+9.
Figure S9. 1H NMR spectrum (500 MHz) of compounds 10+11+12.

S6
Other Chemical Constituents Isolated from Solanum crinitum Lam. (Solanaceae)
J. Braz. Chem. Soc.
Figure S10. 1H NMR spectrum (500 MHz, Pyridine-d5) of the compound 13.
Figure S11. 13C NMR (125 MHz) of compound 14.

Vol. 21, No. 12, 2010
Cornelius et al.
S7
Figure S12. 13C NMR spectrum (125 MHz) of compound 15.
Figure S13. 13C NMR spectrum BBD (A) and DEPT-135 ((B and C), 50 MHz, pyridine-d6) of 16.

S8
Other Chemical Constituents Isolated from Solanum crinitum Lam. (Solanaceae)
J. Braz. Chem. Soc.
Figure S14. 1H NMR spectrum (a) and expansion (b) (400 MHz, CDCl3) of 17.
Molecular Modeling
MP calculations, and is more efficient than conventional
ab initio correlated methods for larger-scale calculations.
The possibility of existence of this epimer as a stable
species was verified by a molecular modeling study at the
DFT B3LYP/6-31G* level with models of both epimers.
The b-epimer model at C-20 (14a) is less stable than the
a-epimer one (13a), but the energy difference between
both structures is only 4.85 kcal mol-1 (20.30 kJ mol-1), as
calculated with the B3LYP/6-31G* DFT method. Because
of this small energy difference, the b-epimer is expected to
exist in appreciable amount in an equilibrium mixture with
the more stable a-epimer. The main reason for the lower
stability of the b-epimer should be the proximity between
the C-21 methyl group and carbons C-23 and C-18. The
corresponding C-C distances, which are equal to 4.22 Å
and 3.47Å, respectively, in 13a, are considerably shorter in
14a, 3.43Å and 2.98Å, respectively. This closer proximity
would raise more unfavorable steric interactions in 14a than
in 13a (Figure S15).
In order to compare the relative stabilities of the
epimeric structures 13/14, a molecular modeling study
was implemented using the Spartan 06 for Linux program
(Wavefunction, Inc.). Because the long chain attached to
C-3 should only have a small influence on the relative
stabilities, it was replaced by a methoxy group to reduce
the computational cost for the calculations. The conformer
distribution of the resulting alpha and beta epimer models
(13a and 14a) was determined with the Monte Carlo
approach using the MMFF molecular mechanics force
field. The most stable conformers of 13a and 14a were
submitted to a previous energy minimization with the PM3
semi empirical method.31 The PM3 optimized structures
were then submitted to a complete energy minimization
with the B3LYP/6-31G* DFT method. The B3LYP method
was chosen because it usually yields results for many
properties in close agreement with those obtained from

Vol. 21, No. 12, 2010
Cornelius et al.
S9
Figure S15. 3D representation of models 13a and 14a after optimization with the B3LYP/6-31G* DFT method.