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

Article
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 and Raimundo Braz-Filhoc
a
Departamento de Química, ICE, Universidade Federal Rural do Janeiro, BR 465 km 07,
23890-000 Seropédica-RJ, Brazil
Instituto de Química, Universidade Estadual de Campinas, 13084-862 Campinas-SP, Brazil
b

c
Setor 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
Solanum (L) is the most representative genus of material for anabolic, anti-fertility, anti-inflammatory,
Solanaceae, containing about 1,500 species, and 5,000 anti-allergic drugs.5 As the alkaloids, the flavonoids are
epithets. It is widespread in tropical and subtropical a frequent group of compounds in Solanum species, and
regions of all the world, but its highest diversity occurs can be used as systematic markers for the family taxons.6-8
in South America.1 Solanum species are a rich source of Solanum crinitum Lam (Solanaceae), popularly known
steroidal alkaloids, flavonoids and their glycosides which as “jurubeba” and “fruto-de-lobo”, is a shrub or small
are known to possess a variety of biological activities. tree in South America, including southern Brazil and
The glycoalkaloids are natural toxins with ecological and Colombia.9,10 In the course of our pharmacological and
human health importance, such as the allelopathic effect phytochemical investigations of Solanum species,3,4,8,11-13
we have reported the cytotoxic and antitumoral activities
*e-mail: [email protected] of flavonoids isolated from trichomes of young branches
2212 Other Chemical Constituents Isolated from Solanum crinitum Lam. (Solanaceae) J. Braz. Chem. Soc.

and also of a rich glycoalkaloids total fraction from the 20-epi-solamargine (14) and solasonine (16) isolated
green fruits of Solanum crinitum Lam14 along with its from the green fruits. The derivatives 3,5,7,4’-tetra-O-
allelopathic activity.3 methyl-kaempferol (4), 3,7,4’-tri-O-methyl-kaempferol
This work describes the isolation and structural (5), 3,7,4’-tri-O-methyl-5-O-acetyl-kaempferol (6),
characterization of the flavonoids tiliroside (1), peracetyl-epi-solamargine (15) and peracetyl-solasonine
astragalin (2), kaempferol (3) and biochanin A-7-O-b- (17) were prepared and used to confirm the structures of
D-apiofuranosyl-(1→5)-b-D-apiofuranosyl-(1→6)-b-D- the natural compounds. The structures were established
glucopyranoside (7), cis- (10) and trans- (11) cumaric through analysis of their spectral data, mainly NMR
acids and ethyl cis- (8) and trans- (9) cumarate and (1D and 2D) and mass spectra. NMR data for the new
4-hydroxybenzoic acid (12), which were isolated from acetyl derivatives of the alkaloids are described herein
the trichomes, and the glycoalkaloids solamargine (13), for the first time.
Vol. 21, No. 12, 2010 Cornelius et al. 2213

Results and Discussion data.21-25 The integration of the 1H NMR signals allowed
us to calculate the approximate relative percentage of both
From the trichomes isolated from green fruits of Solanum compounds: 36.16% of 8 and 63.84% of 9.
crinitum Lam were isolated the flavonoids: tiliroside (1), The fractions containing compounds 13, 14 and 16
astragalin (2), kaempferol (3), and biochanin A-7-O-b- showed a positive test for alkaloids. The detailed analysis
D -apiofuranosyl-(1→5)-b-D -apiofuranosyl-(1→6)-b-D - of the 1H and 13C NMR spectra of the isolated compounds
glucopyranoside (7), cis- (10) and trans- (11) cumaric allowed to identify characteristic signals corresponding to
acids and ethyl cis- (8) and trans- (9) cumarate esters and the same aglycone as that of the steroidal spirazolane-type
4-hydroxybenzoic acid (12); from green fruits methanolic alkaloid in the three glycoalkaloids: four quaternary carbon
extracts were isolated three tri-glycosyl-steroidal alkaloids, atoms, including one linked to oxygen and nitrogen atoms:
solamargine (13), 20-epi-solamargine (14) and solasonine (dC 98.7 to 98.5 and one sp2 at dC 141.2 to 140.4), nine
(16). The derivatives 3,5,7,4’-tetramethylkaempferol methine groups (including two oxygenated at dC 78.5 to 78.0
(4), 3,7,4’-trimethylkaempferol (5), 3,7,4’-trimethyl-5- and 81.1 to 78.6), ten methylene groups and four methyl
acetylkaempferol (6), the peracetyl-20-epi-solamargine groups = (C)3(O-C-NH)(CH)7 (HC-O)2(CH2)10(CH3)4 =
(15) and peracetyl-solasonine (17) were prepared. The C27H42NO3 = C27H42NO2 considering the presence of an
structures were established on the basis of IR, NMR and ether function (Table 1) having a trisaccharide moiety
MS data analysis of the natural compounds and of the (three anomeric carbon atoms: dC 100.6 to 100.3) attached
derivatives 4-6, 15 and 17. to the oxygen atom of carbon CH-3, with dC 78.5 to 78.0
Compounds 1-3 were identified by the analysis of (Table 1 and 2), which is significantly higher when
1
H and 13C NMR spectra, including HMQC and HMBC compared with the 13C chemical shift of the methyl carbon
experiments, and comparison with literature data for CH-3 sustaining free hydroxyl group (about d C 71).
tiliroside, kaempferol and astragalin.15-18 The first report The 1H and 13C chemical shifts of the trisaccharide
of 1 in Solanum species was made by Souza et al.14 moieties of 13 and 14, had practically the same values
Kaempferol and astragalin have been isolated from some (Table 2), indicating identical the partial structure O-[a-
Solanum species.8 The treatment of 3 with diazomethane L-rhamnopyranosyl-(1→2)-O-[a-L-rhamnopyranosyl-
yielded the methyl derivatives 4 and 5, described in the (1→4)]-b-D-glucopyranoside = C18H32O13 (three degrees
literature,19 and the derivative 6 was obtained by treating of unsaturation, C18H38O13 – C18H32O13 = H6). On the other
5 with Ac2O/Pyridine. The 2D NMR spectra, including hand, the 1H and 13C signals observed in NMR spectra of
NOESY experiments, of these derivatives were used to 16 allowed to characterize the trisaccharide as O-[a-L-
confirm the proposed structure of 3 and to carry out the rhamnopyranosyl-(1→2)-O-[b-glucopyranosyl-(1→3)]-
complete assignment of the 1H and 13C chemical shifts of b-D -galactopyranoside = C18H32O14 (three degrees of
6 (see Experimental). unsaturation, C18H38O14 – C18H32O14 = H6). All these NMR
The 1H and 13C NMR spectra of compound 7 revealed spectral data allowed to deduce the same molecular formulas
characteristic resonances of the isoflavonoid biochanin C45H73NO15 to 13 and 14 and C45H73NO16 to 16, all with
A besides additional signals for three sugars unities, one ten degrees of unsaturation corresponding to one double
glucopyranoside and two apiofuranosides. Comparison bond and six rings). In fact, the values of pseudomolecular
of these data with those of glycosides isolated from peaks in the positive HRMS spectra at m/z 868.5239
Dalbergia nigra20 and Andira anthelmia,21 besides the (M + H•+, 100%) of 13 (C45H74NO15 = m/z 868.5058) and
analysis of mass spectrum obtained by FAB-MS in at m/z 868.5305 (M + H•+, 100%) of 14 (C45H74NO15 = m/z
positive mode {m/z 733.20820 ([M+Na]+, C32H38O18+Na+, 868.5058), besides additional peaks compatible with loss
calc.: m/z 733.19558), m/z 601.12300 [M+Na-132]+, m/z of sugar moieties, were used to confirm the molecular
451.0600 (M+Na-282)+, m/z 449.27710 [M+Na-284]+, m/z formula of these isomeric compounds (13 and 14). The
431.16210 [M+Na-302]+, m/z 317.1338 [M+Na-416]+, m/z hydrogen and carbon atoms signals observed in the 1H
287.0865 [M+Na-446]+}, led us to identify the compound and 13C NMR spectra of 13-17 (Tables 1 and 3) were also
as biochanin A, 7-O-b- D -apiofuranosyl-(1→5)-b- D - assigned with aid of the homonuclear 2D 1H-1H-COSY and
apiofuranosyl-(1→6)-b-D-glucopyranoside (7). This is heteronuclear 2D HMQC (1H-13C-COSY-1JCH) and HMBC
the first report of this apiofuranosyl derivative in Solanum (1H-13C-COSY-nJCH, n = 2 and 3), allowing to identify the 1H
species. chemical shifts of the methyl group signals of the aglycone
Compounds 8-12 were identified by the 1H and and of the trisaccharide moieties (Table 3 and 4): dH 0.83
13
C NMR spectral data analysis of the mixtures of 8+9, (d, 3H‑27), 0.88 (s, 3H-18), 1.07 (s, 3H-19) and 1.09 (d,
10+11 and of 12 along with comparison with literature 3H‑21) of 13, 0.79 (d, 3H-27), 0.85 (s, 3H-18), 1.06 (s,
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 shift of the 22bN stereoisomer (Figure 1). The b-position
(s, 3H-18), 1.03 (s, 3H-19) and 1.17 (d, 3H-21) of 16; signals of the methyl Me-21 located at carbon atom 20 of 14 may
of the hydrogen H-6 linked to sp2 carbons of double bounds be used to justify the 13C chemical shifts of the CH-16
at dH: 5.32 (13), 5.33 (14), 5.30 (16), besides the signals of (dC 81.1, absence of the g-effect from Me-21) and CH2-23
H-16 located in the spyrazolane ring (d, J 6 Hz) at dH 4.42 (dC 34.0, presence of the g-effect from Me-21).26-30 In order
(13); 4.82 (14) and 4.21 (16). The additional doublets to compare the relative stabilities of the epimeric structures
(J 6 Hz) corresponding to methyl groups were revealed at 13/14, a molecular modeling study was implemented using
d3H 1.64 and d3H 1.78 in the spectra of 13 and 14 suggested the Spartan 06 for Linux program (Wavefunction, Inc.), see
the presence of two rhamnose moieties in both compounds Supplementary Information.31
and only one rhamnose for 16 as indicated by signal at The complete 1H and 13C chemical shift assignments
dH 1.78. The differences observed in the 1H NMR spectra of the signals of CH3, CH2, CH and C observed in the 1H
of 13 and 14, recorded in same apparatus (1H: 500 MHz; (including 1H-1H-COSY) and 13C ({1H} and DEPT) NMR
13
C: 125 MHz) and solvent pyridine-d5, involving mainly spectra (including 2D experiments HMQC and HMBC)
the chemical shifts of the hydrogen atoms H-16 (dH 4.42 (Tables 1, 2, 3 and 4) and comparison with values described
and 4.82), H-17 (dH 1.78 and 2.02), H-20 (dH 1.98 and 2.14), in the literature for solamargine (13)26-29 and solasonine
2H-26 (dH 2.78 and 3.04/2.89) and 3H-21 (dH 1.09 and (16)28,31 led to the proposition of the structures (25R)
1.28), which revealed correlation in the HMQC with the 13C (20S)(16S)-3b-{O-a-L-rhamnopyranosyl-(1→2)-O-[a-L-
signals of the corresponding carbon atoms: CH-16 (dC 79.2 rhamnopyranosyl-(1→4)]-b-D -glucopyranosyl}-22aN-
and 81.1), CH-17 (dC 63.9 and 63.0), CH-20 (dC 42.0 and spirosol-5-en (solamargine, 13), the new glycoalkaloid
42.0), CH2-26 (dC 48.2 and 47.1) and 3H-21 (dC 16.1 and (25R)(20R)(16S)-3b-{O-a-L-rhamnopyranosyl-(1→2)-
15.7), as summarized in Figure 1. Comparative analysis O-[a-L-rhamnopyranosyl-(1→4)]-b-D-glucopyranosyl}-
of these data was used to suggest the two stereoisomers 22aN-spirosol-5-en (16-epi-solamargine, 14), and (25R)
H-20b (13) and H-20a (14), since the 22aN and 22bN (20S)(16S)-3b-{O-a-L-rhamnopyranosyl-(1→2)-O-[b-
possibilities were eliminated considering the absence of glucopyranosyl-(1→3)]-b-D-galactopyranosyl} -22aN-
13
C signal corresponding to a methylene carbon CH2-23 at spirosol-5-en (solasonine, 16) for the three steroidal
about dC 27 which is consistent with the 13C NMR chemical glycoalkaloids isolated from this plant (Figure 1).

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 Experimental
(125 MHz: 13 and 14; 50 MHz: 16) and of peracetyl derivatives 15 and
17 in CDCl3 (125 MHz)*
General procedure
C 13 14 15a 16 17a
Melting points have not been corrected. IR, NMR and
1 (CH2) 37.9 37.5 37.3 37.6 37.1
mass spectra were recorded on the same equipments used
2 (CH2) 30.5 30.2 29.7 30.2 29.5 in previous papers.21,32 Column chromatography was carried
3 (CH) 78.5 78.0 79.3 78.4 79.8 out with silica gel (Vetec and Aldrich 0.05-0.20 mm) and
4 (CH2) 39.3 39.0 38.4 38.8 38.4 Sephadex LH-20 (Sigma, USA); silica gel F254 G (Vetec)
5 (C) 141.2 140.4 140.4 141.0 140.2
was used for preparative TLC; aluminum backed (Sorbent)
silica gel plates W/UV254 were used for analytical TLC,
6 (CH) 122.2 121.9 122.2 121.8 121.9
with visualization under UV (254 and 366 nm), with AlCl3-
7 (CH2) 32.1 32.3 31.8 32.3 31.6 ETOH (1%), Liebermann-Burchard and/or Godin reagents,
8 (CH) 32.0 31.7 31.6 31.8 31.4 or exposure to iodine vapor.
9 (CH) 50.7 50.3 50.3 50.3 50.0
10 (C) 37.5 37.2 36.9 37.2 36.7 Plant material
11 (CH2) 21.5 21.1 21.0 21.2 20.6
The green fruits of Solanum crinitum Lam. were
12 (CH2) 40.5 39.7 40.0 39.9 39.8
collected in September, 2001, in the campus of Universidade
13 (C) 41.0 40.8 42.9 40.8 42.7 Federal Rural do Rio de Janeiro (UFRRJ), Seropédica-RJ,
14 (CH) 57.0 56.5 53.0 56.6 54.8 Brazil. They were collected by M.Sc. José Milton Alves
15 (CH2) 32.9 32.6 31.6 32.6 31.6 (Agronomy Institute, UFRRJ). The identification was made
16 (CH) 79.2 81.1 75.9 78.6 75.6
by Dr. Maria de Fátima Agra, Laboratório de Tecnologia
Farmacêutica, Universidade Federal da Paraíba, João
17 (CH) 63.9 63.0 62.1 63.2 62.0
Pessoa-PB. A voucher specimen (No. JP-28000) was
18 (CH3) 16.9 16.4 12.8 16.5 12.4
deposited at the Herbarium Prof. Lauro Pires Xavier,
19 (CH3) 19.8 19.4 19.4 19.5 20.3 Universidade Federal da Paraíba, João Pessoa-PB, Brazil.
20 (CH) 42.0 42.0 40.0 41.9 39.7
21 (CH3) 16.1 15.7 21.1 15.8 21.4 Extraction and isolation
22 (C) 98.7 98.5 99.7 98.5 99.2
The trichomes (9.7 g) were isolated by scraping the
23 (CH2) 35.0 34.0 35.2 34.3 35.0
green fruits with a glass slide and were subsequently
24 (CH2) 31.4 30.0 30.1 30.5 30.8 extracted with CHCl3 and MeOH in an ultrasound bath to
25 (CH) 32.1 31.7 29.1 30.8 28.9 furnish the CHCl3 extract (310.0 mg) and MeOH (3.0 g)
26 (CH2) 48.2 47.1 45.5 47.5 45.2 residues. The CHCl3 residue was chromatographed on a
27 (CH3) 20.1 19.2 18.5 19.5 19.2
sephadex CC, using MeOH as eluent, and 13 fractions were
collected and analyzed by TLC and 1H NMR spectroscopy.
* Number of hydrogens bound to carbon atoms deduced by comparative
analysis of {1H}- and DEPT-13C NMR spectra. 2D 1H-1H-COSY and Fractions 7-8 yielded astragaline (1, 10.0 mg) and fractions
1
H-13C-nJCH (n = 2 and 3) NMR spectra were also used in these assignments; 9-10 yielded the tilirozide (2, 25.0 mg). The methanolic
a
dCOCH3: 170.1-170.2 (CO), 23.8 (CH3). extract was chromatographed on a silica gel column (col
A) and 20 fractions were collected and analyzed by TLC
The natural alkaloids 14 and 16 were treated with plate. Fractions 5-8 (2.0 g) were dissolved in methanol and
Ac2O/pyridine to yield the peracetyl derivatives 15 and addition of CH2Cl2 yielded a precipitate that was separated
17. The 1H and 13C NMR spectral data of these peracetyl by filtration to afford a solid (1.6 g) that was identified
derivatives (15 and 17), obtained through the analysis of as kaempferol (3, mp 282-284 oC) and the mother liquor
extensive 1D and 2D NMR experiments (Tables 1-4), were (AM-5-8). The reaction of kaempferol (65.0 mg) with
also used to confirm the postulated structures to 14 and 16. diazomethane yielded 16.1 mg of 4 (mp 128-130 oC) and
The analysis of 1D and 2D NMR spectra was also used to 36.6 mg of 5 (mp 140-142 °C) that were separated by
make the complete hydrogen and carbon-13 chemical shift preparative TLC. 20.3 mg of 5 were treated with Ac2O/
assignments for the alkaloid, the new 16-epi-solamargine pyridine (1:1) to afford 6 (14.8 mg, mp 166-168 °C). The
(14) and for the two peracetyl derivatives 15 and 17. 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 trisaccharide of 14 trisaccharide of 15a trisaccharide of 16 trisaccharide of 17a

1’ (CH) 100.6 100.3 99.8 100.4 99.6
2’ (CH) 78.2 77.8 76.5 76.6 73.1
3’ (CH) 78.3 78.1 75.5 84.9 78.3
4’ (CH) 79.0 78.8 77.9 70.5 69.5
5’ (CH) 77.3 76.9 72.5 77.5 70.9
6’ (CH2) 61.7 61.3 62.1 62.6 62.0
1” (CH) 103.3 102.9 99.7 105.9 99.2
2” (CH) 72.9 72.5 70.5 72.6 71.8
3” (CH) 73.1 72.8 68.8 80.2 72.3
4” (CH) 74.3 73.9 70.8 75.2 68.2
5” (CH) 70.8 70.5 68.1 77.5 72.0
6” (CH3) 18.9 18.5 17.4 62.6 60.9
1’’’ (CH) 102.4 102.1 97.5 102.3 97.0
2’’’ (CH) 72.9 72.6 70.2 72.9 68.9
3’’’ (CH) 73.1 72.9 68.8 74.2 69.1
4’’’ (CH) 74.5 74.2 71.5 75.0 70.9
5’’’ (CH) 69.9 69.5 66.7 69.5 66.5
6’’’ (CH3) 19.0 18.7 17.5 18.7 17.1
* Number of hydrogens bound to carbon atoms deduced by comparative analysis of { H}- and DEPT- C NMR spectra. 2D H- H-COSY and 1H-13C-nJCH
1 13 1 1

(n = 2 and 3) NMR spectra were also used in these assignments; a dCOCH3: 169.9-170.9 (CO), 20.9-21.7 (CH3).

gel CC and fractions 8-12 yielded a mixture of 8 and 9 10 (8.67 g) was extracted with methanol and the alkaloids
(1.2 mg, gum). Fraction 10 (850 mg) from col A was were detected with Dragendorff and Libermann Burchard.
chromatographed on a silica gel column and 130 fractions This fraction was chromatographed on a silica gel column
of 50 mL were collected. Fractions 32-40 yielded 35.5 mg using CH2Cl2/MeOH (3:1) as initial eluent. 79 fractions were
of a material corresponding to the mixture of 10, 11 and 12. collected and were analyzed by TLC plate and reunited in
Fractions 94-114 (102.3 mg) were chromatographed on a group. Fractions 24-29 (1.2 g) were filtered on a Sephadex
silica gel column and fractions 3-5 yielded 7 (9.5 mg) after LH-20 column and the epi-solamargine (14, 225.8 mg,
filtration on Sephadex LH20. 238-250 °C) was obtained. Acetic anhydride:pyridine (1:1)
The green fruits (2.6 g) of Solanum crinitum Lam were treatment of 14 (44.5 mg) yielded 44.5 mg of the peracetyl
powdered and extracted with ethanol + acetic acid (2%) and derivative 15 (mp 128-130 °C). Fractions 39-44 (1.3 g)
900 mL of solution was obtained. 900 mL of acetic acid were filtered on Sephadex LH20 to isolate solasonine (16,
(10%) were added, and the solution was left to stand and 93.5 mg, mp 244-246 °C). Treatment of 16 (79.1 mg) with
chill overnight. The solution was filtered under vacuum using acetic anhydride:pyridine (1:1) gave the peracetyl-solasonine
Hirsch funnel. NH4OH (pH 9-10) was added to the filtrate (17, 27.6 mg, mp 148-150 °C).
which was allowed to stand overnight in the fridge affording
a precipitate (96.4 g) corresponding to the glycoalkaloids 3,7,4’-trimethoxy-5-acethoxyflavone (6)
fraction was obtained. 90 g of the glycoalkaloids was Yellow powder, mp 166-168 °C; IR (KBr) nmax/cm-1:
adsorbed on silica gel and applied on a silica gel column, 1762, 1632, 1606, 1511; 1H NMR (CDCl3, 500 MHz) d
eluted with hexane, dichloromethane, ethyl acetate, acetone 6.60 (d, J 2.5 Hz), 6.82 (d, J 2.5 Hz), 8.02 (dd, J 9.0 Hz),
and methanol. 14 fractions of 500 mL were collected. Fraction 7.02 (d, J 9.0 Hz), 3.78, 3.90 (s), 3.89 (s), 2.47 (s), 13C NMR
8 (1.0 g), collected with ethyl acetate, was chromatographed (CDCl3, 125 MHz) d 154.8 (C-2), 140.8 (C-3), 173.2 (C-4),
in a silica gel CC and fractions analyzed by silica gel TLC 150.5 (C-5), 108.0 (CH-6), 163.1 (C-7), 98.6 (CH-8), 157.8
plate. Fraction 84 (52.6 mg) from this column was filtered (C-9), 111.4 (C-10), 122.9 (C-1’), 129.9 (CH-2’,6’), 114.0
on a Sephadex LH20 column, eluted with methanol, and the (CH-3’,5’), 161.4 (C-4’), 59.9, 55.9, 55.3 (MeO-3,7,4’,
alkaloid solamargine 13 (268-270 °C) was obtained. Fraction 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 13 14 15a 16 17a
1 1.73/1.00 1.75/1.02 1.85/1.08 1.85/1.08
2 2.10/1.87 2.08/1.85 1.93/1.58 1.95/1.25
3 3.89 (m) 3.89 (m) 3.60 3.93 (m) 3.58 (m)
4 2.82/2.75 2.81 (dd, J 11.6; 3.1) 2.40/2.25 2.87/2.76 2.45/2.28
2.73 (t, J 11.6 )
6 5.32 (br s) 5.33 (br s) 5.36 (m) 5.30 (br s) 5.38 (m)
7 1.90 1.85 1.94 2.30/1.68
8 1.48 1.58 1.56 1.53
9 0.90 0.89(m) 0.95 0.95
11 1.45 1.42 1.55-1.45 1.55-1.45
12 1.63/1.05 1.68/1.09 2.08/1.25 2.08/1.25
14 1.10 1.09 (m) 0.98 0.98
15 2.08/1.45 2.08/1.48 2.20/1.70 1.95/1.85
16 4.42 4.82 5.06 4.21 5.08
17 1.78 2.02 1.53 1.53
18 0.88 (s) 0.85 (s) 0.87 (s) 0.82 (s) 0.91 (s)
19 1.07 (s) 1.06 (s) 1.01 (s) 1.03 (s) 1.04 (s)
20 1.98 2.14 2.10 2.08
21 1.09 (d, J 6.9) 1.28 (d, J 6.7) 1.26 (d, J 6.9) 1.17 (d) 1.26 (d, J 7.0)
23 1.90/1.65 1.92/1.78 2.36/1.04 2.35/1.03
24 1.52 1.65 2.28/1.68 1.70/1.65
25 1.65 1.58 2.02 2.02
26 2.78 3.04 (m)/2.89 (t, J 11.5) 3.20 2.83 3.16(m)
27 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 dCOCH3: 2.16 (s).

Epi-solamargine (14) C63H92NO24: 1246.600928), m/z 1204.5278 (calc. for

Colorless crystals, mp 238-250 °C; [a]25D – 98 (c 0.1, C61H90NO23: 1204.590364) [M+H-42]+, m/z 974.5256
MeOH); IR (KBr) nmax/cm-1: 3417, 2927, 1453, 1384, (calc. for C51H76NO17: 974.511325) [M+H-rhamnose]+, m/z
1044 cm-1. HR-FABMS positive ions at m/z 868.5239 932.5163 (calc. for C49H74NO16: 932.500760) [M-314]+, m/z
[M+H]+ (calc. for C45H74NO15: m/z 868.505846), 722.4643 634.7963 (calc. for C39H56NO6: 634.410763) [M+H‑612]+,
(calc. for C39H64NO11: 722.447937) [M+H-rhamnose]+, m/z 413.2715 (calc. for C 27 H 43 NO 2 : 413.329379)
m/z 576.4037 (calc. for C33H54NO7: 576.390028) [M+H- [M-acetate-rhamnose-rhamnose-glucose] +. 13C RMN:
rhamnose-rhamnose]+, m/z 445.7553 (calc. for C21H33O10: Tables 1 and 2. 1H RMN: Tables 3 and 4.
445.207372) [M-423]+, m/z 414.3467 (calc. for C27H44NO2:
414.337204) [M+H-rhamnose-rhamnose-glucose]+, m/z Peracetyl-solasonine (17)
413.2782 (calc. for C27H43NO2: 413.329379) [M-rhamnose- Crystal, mp 148-150 °C; 13C RMN: Tables 1 and 2;
rhamnose-glucose]+. 13C RMN: Tables 1 and 2. 1H RMN: 1
H RMN: Tables 3 and 4.
Tables 3 and 4.
Supplementary Information
Peracetyl-epi-solamargine (15)
Crystal, mp 128-130 °C, [a]25D – 165 (c 0.1, MeOH); Supplementary data associated with this paper are
IR (KBr) nmax/cm-1: 2943. 2728, 1751, 1438, 1374, 1045; available free of charge at http://jbcs.sbq.org.br, as a PDF
Positive ion FABMS m/z 1246.5522 [M+H]+ (calc. for 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 Glycoside unit of 14 Glycoside unit of 15a Glycoside unit of 16 Glycoside unit of 17a
1’ 4.95 4.96 4.56 (d, J 7.7) 4.95 4.46 (d, J 8.0)
2’ 4.24 4.22 3.56 3.98 3.84 (dd, J 8.0; 9.5)
3’ 4.23 (t) 4.23 5.28 (t, J 9.4) 4.33 (m) 3.91 (dd, J 9.5; 3.0)
4’ 4.42 (t) 4.40 3.73 (t, J 9.4) 4.82 5.26 (br s)
5’ 3.65 (br d, J 9.2) 3.66 (td, J 9.2) 3.59 (m) 3.79 (m)
6’ 4.22(br d, J 12.0) 4.23 (dd, J 12.1; 3.2)n 4.44 (d, J 12.3) 4.12 (m)
4.10 (br d, J 12.0) 4.10 (dd, J 12.1; 3.2) 4.28 (dd, J 12.3; 3.6) 4.06 (m)
1” 5.86 (s) 5.87 (s) 4.79 (d, J 1.5) 5.17(d) 4.71 (d, J 8.0)
2” 4.69 (sl) 4.69(d, J 1.4) 5.02 (m) 4.68 (sl) 4.85 (dd)
3” 4.55 (dd, J 9.1; 2.9) 4.55 (dd, J 9.2; 3.2) 5.16 (dd, J 10.2; 3.1) 5.22 (t)
4” 4.38 4.34 (t, J 9.2) 5.03 (t, J 10.2) 5.08
5” 4.92 4.93 (m) 3.85 (m) 3.69 (m)
6” 1.64 (d, J 5.9) 1.64 (d, J 6.1) 1.14 (d, J 6.1) 4.38 (br d, J 10.5)/4.10
1’’’ 6.41 (s) 6.41 (s) 4.90 (d, J 1.5) 6.28 (s) 5.02 (sl)
2’’’ 4.84 (br s) 4.84 (d, J 1.8) 5.00 (m) 4.90 (d) 5.22
3’’’ 4.64 (dd, J 9.1; 2.9 4.64 (dd, J 9.2; 3.3) 5.23 (dd, J 10.1 Hz; 3.1) 4.61 5.22
4’’’ 4.34 4.32 (t, J 9.2) 5.05 (t, J 10.1) 5.02(m)
5’’’ 4.96 4.98 (m) 4.36 (m) 4.88 (br s) 4.43 (m)
6’’’ 1.78 (d, J 6.0) 1.78 (d, J 6.2) 1.17 (d, J 6.2) 1.78 (d) 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 dCOCH3: 1.98-2.15 (8×CH3).

compare the relative stabilities of the epimeric structures 5. Mola, J. L.; Araujo, E. R.; Magalhães, G. C.; Quim. Nova 1997,
13 and 14. 20, 460.
6. D’arcy, W. G. In The Biology and Taxonomy of the Solanaceae;
Acknowledgments Hawkes, J. G.; Lester, R. N.; Skelding, A., eds., Academic Press:
London, 1979, pp. 3-49.
The authors are grateful to CNPq, FAPERJ and 7. Steinharter, T. P.; Cooper-Driver, G. A.; Anderson, G. J.;
CAPES for research fellowships and financial support. Biochem. Syst. Ecol. 1986, 14, 299.
They also thank CENAUREMN for the 500 MHz NMR 8. Silva, T. M. S.; Carvalho, M. G. de; Braz-Filho, R.; Agra, M.
spectra, Laboratório Plataforma de Métodos Analíticos de F.; Quim. Nova 2003, 26, 517.
Farmanguinhos/FIOCRUZ for the 400 MHz NMR spectra. 9. Agra, M. F.; PhD Thesis, Universidade de São Paulo, Brazil, 2000.
They thank MSc J.M. Alves and Dr. M. de F. Agra for the 10. Dias-Filho, M. B.; Pesq. Agropec. Bras. 1997, 32, 789.
collection and classification of plant material. 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.
References Syst. Ecol. 2004, 32, 513.
12. Silva, T. M. S. da; Braz-Filho, R.; Carvalho, M. G. de; Agra,
1. Agra, M. F.; Novon 1999, 9, 292. M. de F.; Biochem. Syst. Ecol. 2002, 30, 1083.
2. Fukuhara, K.; Shimizu, K.; Kubo, I.; Phytochemistry 2004, 65, 13. Silva, T. M. S. da; Silva, C. de C.; Braz-Filho, R.; Carvalho, M.
1283. G. de; Agra, M. de F.; Rev. Bras. Farmacogn. 2002, 12, 85.
3. Alves, C. C. F.; Alves, J. M.; Silva, T. M. S.; Carvalho, M. G. 14. Souza, A. E.; Silva, T. M. S. da; Alves, C. C. F.; Carvalho, M.
de; Neto, J. J.; Floresta e Ambiente 2003, 10, 93. G. de; Braz-Filho, R.; Echevarria, A.; J. Braz. Chem. Soc. 2002,
4. Silva, T. M. S.; Câmara, C. A.; Agra, M. F.; Carvalho, M. G. 13, 838.
de; Frana, M. T.; Brandoline, S. V. P. B.; Paschoal, L. S.; Braz- 15. Markham, K. R. In Methods in Plant Biochemistry; Harborne,
Filho, R.; Fitoterapia 2006, 77, 449. 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.
17. Agrawal, P. K.; Bansal, M. C. In Carbon-13 NMR of Flavonoids, 26. Ripperger, H.; Phytochemistry 1995, 39, 1475.
Studies in Organic Chemistry; Elsevier: Amsterdam, 1989, p. 362. 27. Lorey, S.; Porzel, A.; Ripperger, H.; Phytochemistry 1996, 41,
18. Markham, K. R.; Geiger, H. In The Flavonoids Advances in 1633.
Research Since 1986; Harborne, J. B., ed., Chapman & Hall: 28. Wanyonyi, A. W.; Chiabra, S. C.; Mkoji, G.; Eilert, U.; Njue,
London, 1994, pp. 441. W. M.; Phytochemistry 2002, 59, 79.
19. Dong, H. Y.; Gou, L.; Cao, S. G.; Chen, S. X.; Sim, K. Y.; Goh, 29. Weissenberg, M.; Phytochemistry 2001, 58, 501.
S. H.; Kini, R. M.; Phytochemistry 1999, 50, 899. 30. Usubillaga, A.; Aziz, I.; Tettamanzi, M. C.; Waibel, R.;
20. Mathias, L.; Vieira, I. J. C.; Braz-Filho, R.; Rodrigues-Filho, Phytochemistry 1997, 44, 537.
E. A.; J. Nat. Prod. 1998, 61, 1158. 31. Lee, C.; Yang, W.; Parr, R. G.; Phys. Rev. B: Condens. Matter
21. Silva, V. C. da; Carvalho, M. G. de; Silva, S. L. da C. e; Rev. Mater. Phys. 1988, 37, 785.
Latinoam. Quim. 2007, 35, 13. 32. Carvalho, M. G. de; Silva, V. C. da; Silva, T. M. S. da; Câmara,
22. Marin, J. C.; Torres, F.; Rev. Lationoam. Quim. 2001, 29, 100. C. A.; Braz-Filho, R.; An. Acad. Bras. Cienc. 2009, 81, 21.
23. Rasmussen, S.; Wolff, C.; Rudolph, H.; Phytochemistry 1996,
42, 81. Submitted: March 23, 2009
24. Silva, A. M. S.; Alkorta, I.; Elguero, J.; Silva, V. L. M.; J. Mol. Published online: August 12, 2010
Struct. 2001, 595, 1.
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

Supplementary Information
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 and Raimundo Braz-Filhoc
a
Departamento de Química, ICE, Universidade Federal Rural do Janeiro, BR 465 km 07,
23890-000 Seropédica-RJ, Brazil
Instituto de Química, Universidade Estadual de Campinas, 13084-862 Campinas-SP, Brazil
b

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