J. Braz. Chem. Soc., Vol. 18, No. 6, 1132-1135, 2007. Printed in Brazil - ©2007 Sociedade Brasileira de Química 0103 - 5053 $6.00+0.00
ArticleArticleArticleArticleArticle
*e-mail: [email protected]
New Flavone from the Leaves of
Neea theifera
(Nyctaginaceae)
Daniel Rinaldo, Clenilson M. Rodrigues, Juliana Rodrigues, Miriam Sannomiya,
Lourdes C. dos Santos and Wagner Vilegas*
Instituto de Química, Universidade Estadual Paulista, CP 355, 14801-900 Araraquara-SP, Brazil
Neea theifera Oerst. (Nyctaginaceae) é amplamente utilizada na medicina popular brasileira
para tratamento de úlceras gástricas e inflamação. A investigação fitoquímica das folhas de
Neea theifera permitiu o isolamento e identificação da nova flavona luteolina-7-O-[2''-O
-(5'''-O-feruloil)-β-D-apiofuranosil]-β-D-glucopiranosídeo (1) além dos oito compostos conhecidos vitexina, isovitexina, isoorientina, orientina, vicenina-2, crisoeriol, apigenina e luteolina. A identificação química foi realizada por métodos espectroscópicos incluindo RMN-2D, bem como análises no UV e IES-EM.
Neea theifera Oerst. (Nyctaginaceae) is widely used in Brazilian folk medicine for the
treatment of gastric ulcers and inflammation. Phytochemical investigation of the leaves of
Neea theifera afforded the isolation of the new flavone luteolin-7-O-[2''-O-(5'''-O-feruloyl)-β
-D-apiofuranosyl]-β-D-glucopyranoside (1) besides the eight-known compounds vitexin, isovitexin, isoorientin, orientin, vicenin-2, chrysoeriol, apigenin and luteolin. Their chemical identification was established by NMR spectroscopic methods including 2D-NMR, as well as UV and ESI-MS analyses.
Keywords: Neea theifera, Nyctaginaceae, flavone, luteolin-7-O-[2''-O-(5'''-O-feruloyl)-β -D-apiofuranosyl]-β-D-glucopyranoside
Introduction
The Nyctaginaceae family comprises around 300 species in 30 genera.1 Phytochemical investigation with plants from this family is still scarce. Previous phytochemical studies of Boerhavia coccinea and B. erecta led to the isolation of tannins and saponins,2 while B. diffusa yielded dihydroisofuranoxanthone,3 rotenoids4 and lignans.5Betacianins and flavonoids were isolated from Bougainvillea glabra,6 already flavonoids and phenolic compounds were isolated from B. spectabillis.7,8 Saponins were isolated from Colignonia scandens Benth.9 and from Pisonia umbellifera.10
In our search for bioactive natural products from São Paulo State, Brazil, we have examined the leaves of Neea theiferaOerst. This species is popularly known as ‘Capa-rosa-do-campo’ that grow wild in cerrado lands of Brazil.1 They are used in folk medicine for the treatment of gastric ulcers and inflammation.11 To the best of our knowledge, we could not find any previous phytochemical investigation with plants belonging to this genus.
The present paper describes the isolation, purification and structure elucidation of the nine compounds from the leaves of N. theifera.
Experimental
Plant material
Neea theifera Oerst. was collected in March 2005, at Corumbataí, Itirapina city, São Paulo State, Brazil, and authenticated by Prof. Dr. Jorge Yoshio Tamashiro from the Instituto de Biologia, Unicamp, São Paulo. A voucher specimen (HUEC 1396) is on file of the Herbarium of the Universidade Estadual de Campinas, Brazil.
Extraction and isolation
The dried leaves of N. theifera (953.5 g) were
1133
Rinaldoet al.
Vol. 18, No. 6, 2007
respectively. The fr. 1:1 was chromatographed on
Sephadex LH-20 with H2O:MeOH (1:1) as eluent.
Fractions (4.0 mL) were collected and checked by TLC [Si gel plates, CHCl3:MeOH:n-PrOH:H2O (5:6:1:4), organic phase] giving 426 fractions. Fractions 53-58, 59-61, 66-68, 162-170, 190-206, 233-254 and 284-301 were identified as vitexin (12 mg), isovitexin (26 mg), orientin (16 mg), vicenin-2 (8 mg), chrysoeriol (5 mg), apigenin (4 mg) and luteolin (4 mg), respectively. Fraction 74-82 (60 mg) was further purified by semi-preparative HPLC and afforded to obtain pure isoorientin (6 mg) and pure compound (1) (8 mg).
General experimental procedures
Melting point was measured on a digital MQ APF-301 (Microquímica®, Brazil) apparatus. UV spectrum was recorded on a HACH UV-Vis DR/4000 spectrophotometer in MeOH. IR spectrum was obtained using Shimadzu FT-IR 8300 spectrophotometer in KBr disk. NMR analyses
and 2D experiments were run on Varian® INOVA 500
operating at 500 MHz for 1H and 125 MHz for 13C (11.7 T), using TMS as internal standard. The ESI-MS spectra were obtained with a Fisons Platform spectrometer in negative mode (70 V). The samples were dissolved in MeOH and injected directly. HREIMS was performed by using an ultrOTOFQ-ESI-TOF Mass Spectrometer Bruker Daltonics® instrument. The compound (1) was dissolved in MeOH, mixed with the internal calibrant, and introduced directly into the ion source by direct infusion.
TLC analyses were performed on silica gel 200 µm
(Sorbent Technologies®) and visualized using UV light (254 and 365 nm).
Semi-preparative HPLC analysis
Semi-preparative HPLC analysis was obtained on a HPLC Varian®ProStar 210 system equipped with a Varian® 330 photodiode array detector with a Phenomenex® Luna(2) RP18 column (40 × 2.5 cm , 10 µg) and Reodyne injector of 100 µL.
A binary gradient elution system with solvent A (0.05% TFA in H2O) and solvent B (0.05% TFA in CH3CN) was applied with linear gradient formation initially with 68:32 (A:B) at 35 min, and then it was changed to 55:45 (A:B) at 5 min, and finally 55:45 (A:B) at 30 min. The flow-rate was 4.7 mL min–1.
Luteolin-7-O-[2''-O-(5'''-O-feruloyl)-β-D -apiofuranosyl]-β-D-glucopyranoside
Yellow amorphous powder. mp 250-252 °C. UV
(MeOH)λmax/nm: 206, 249 and 334. IR (KBr) νmax/cm-1: 3433, 1655 and 1605. ESI-MS m/z: 755 [M-H]–, 579 [M-E-feruloyl-H]–, 561 [M-E-feruloyl-H
2O-H]
–, 447 [M-E
-feruloyl-apiose-H]–, 285 [M-E -feruloyl-apiose-glucose-H]–. HRESIMS [M-H]–m/z 755.1902 (calculated for C36H36O18 –H, 755.1823). For 1H and 13C NMR analyses see Table 1.
Results and Discussion
The methanolic extract of the dried powdered leaves of N. theifera was purified by column chromatography (CC) on a Sephadex LH-20 column followed by purification by HPLC-DAD, afforded the isolation of the new compound (1). In addition, eight-known compounds were identified by comparison of their spectroscopic properties with published data: vitexin, isovitexin, isoorientin, orientin, vicenin-2, chrysoeriol, apigenin and luteolin.12-14
Compound (1) was isolated as a yellow solid (mp 250-252 °C). The UV spectral data showed absorption bands at 249 nm and 334 nm. The IR spectrum presented bands at 3433 cm–1 (OH), at 1655 cm–1 (C=O) and 1605 cm–1 (C=C).
The molecular formula of compound (1) was calculated as C36H36O18 from its HRESIMS, which
showed a [M-H]– at m/z 755.1902 (calculated for
C36H36O18-H, 755.1823). The negative ESI-MS (70 V) exhibited the pseudomolecular ion [M-H]–at m/z 755. Key fragmentation ions occurred at m/z 579 [M-E -feruloyl-H]–, m/z 561 [M-E-feruloyl-H
2O-H]
–, m/z 447
[M-E-feruloyl-apiose-H]– and m/z 285 [M-E -feruloyl-apiose-glucose-H]–.
The1H NMR spectrum (Table 1) showed signals at G 7.34 (1H, d, J 8.0 Hz), G 7.35 (1H, brs) and at G 6.88 (1H, d,J 8.0 Hz) assigned to H-6', H-2' and H-5' respectively, two doublets at G 6.68 (1H, d, J 2.0 Hz) and G 6.36 (1H, d,J 2.0 Hz), attributed to H-8 and H-6 of the A-ring, and one singlet at G 6.54 (1H, s) attributed to H-3 typical of a luteolin derivative. Signals at G 6.17 (1H, d, J 16 Hz, H-α),G 7.30 (1H, d, J 16 Hz, H-β),G 7.07 (1H, d, J 1.5 Hz, H-2''''),G 6.69 (1H, d, J 8.0 Hz, H-5'''') and G 6.87 (1H, d, J 8.0 and 1.5 Hz, H-6'''') suggested the presence of an E -feruloyl unit.15 A signal at G 3.74 (3H, s) indicates the
presence of a methoxyl group.13NOESY experiment
showed correlation between signal at G 3.74 (OMe) and atG 7.07, thus establishing the methoxyl group at position 3'''' of the E-feruloyl unit.
1134 New Flavone from the Leaves of Neea theifera (Nyctaginaceae) J. Braz. Chem. Soc.
units. The TOCSY experiment with irradiation at G 5.20 displayed the spin system of the β-D-glucopyranoside unit, whereas irradiation at G 5.38 resulted only in the singlet atG 3.75 (1H, s) suggesting an apiofuranosyl unit. The coupling constant of the anomeric proton at G 5.20 (1H, d,J 7.5 Hz) indicated that the present glucose unit has a β-configuration.13
The13C NMR experiment presented 35 signals, from which 15 were attributed to the aglycone, 9 to the E-feruloyl unit, 6 to the β-D-glucopyranosyl unit, and with 5 was possible determined a apiofuranosyl unit.14
The apiose unit was characterized through 1H and 13C NMR experiments compared to the literature data. In the 1H NMR spectrum, apiose unit with OH linked to C-1'''
and OH linked to C-2''' in trans configuration presents constant coupling J1,2 0-1 Hz, whereas cisconfiguration is characterized by J1,2 3-4 Hz.16,17 The chemical shift of C-1 in 13C NMR experiments in pyridine-d
5 to the α-D
-apiofuranoside is G 105 and G112 to β-D-apiofuranoside,18 whereas in DMSO-d6 these isomers produce signals at G 108 and G109, respectively.19,20,21 Thus, the apiose unit in (1) was identified as being aβ-D-apiofuranoside.
The structure and bonds of these units on compound
(1) was established from gHMQC and gHMBC
experiments. gHMQC experiment showed direct
correlations between carbons and the respective hydrogens
(Table 1). gHMBC experiments showed long-range
correlations between the hydrogen signal at G 5.20 (H-1'’ glucose) and the carbon signal at G 162.4 (C-7 aglycone), and between the hydrogen signal at G 3.52 (H-2'’ glucose) and the carbon signal at G 108.2 (C-1''' apiose). Besides, the chemical shift of the C-2'' of the glucose (G 75.8) unit was clearly dishielded (+3) compared to the chemical shift of the analogous carbon resonance of a non-substituted glucose unit (G 72.9), supporting the glucose (1→2) apiose
linkage.14 The gHMBC experiment also showed
Table 1. NMR spectral data of compound 1 (11.7 T, DMSO-d6)a
Position 1H NMR (G
H)
13-C NMR (G
C) gHMBC (GH)
2 164.4 H-3
3 6.54 s 103.0
4 181.7
5 12.91 161.2 H-6
6 6.36 d (2.0) 99.1 H-8
7 162.4 H-1''; H-6; H-8
8 6.68 d (2.0) 94.4 H-6
9 156.9 H-8
10 105.4 H-6; H-3; H-8
1' 121.6
2' 7.35 brs 113.4 H-6'
3' 149.3 H-5'
4' 145.0 H-2'; H-6'
5' 6.88 d (8.0) 115.9
6' 7.34 d (8.0) 119.1
Glucose
1'' 5.20 d (7.5) 97.8 H-2''
2'' 3.52 t (7.5; 7.5) 75.8
3'' 3.45 t (7.5; 7.5) 76.7 H-2''
4'' 3.20 t (7.5; 7.5) 69.8 H-2''
5'' 3.48 m 77.0 H-4''; H-3''
6'' a) 3.50 m b) 3.71 d (10.0) 60.5
Apiose
1''' 5.38 s 108.2 H-2'''; H-2''
2''' 3.75 s 76.6
3''' 77.5 H-2'''; H-4'''
4''' a) 3.78 d (9.5) b) 4.04 d (9.5) 73.8 H-5'''; H-2'''
5''' 4.06 brs 66.6 H-2'''; H-4'''
E-Feruloyl
α 6.17 d (16.0) 113.7 H-β
β 7.30 d (16.0) 144.9 H-6''''; H-2''''
1'''' 125.4 H-α; H-5''''
2'''' 7.07 d (1.5) 110.9 H-6''''; H-β
3'''' 147.8 -OCH3; H-5''''
4'''' 149.3 H-6''''; H-2''''
5'''' 6.69 d (8.0) 115.4 H-6''''
6'''' 6.87 dd (8.0; 1.5) 122.9 H-2''''; H-β
-OCH3 3.74 s 55.5
C=O 166.3 H-5'''; H-β
aChemical shifts (G) are in ppm, and coupling constants (J in Hz) are
given in parentheses.
Figure 1. Structure of compound (1).
α β
1 4
2 3 5
6
7 8 9
10
1' 2'
3' 4'
5' 6' 1’’
2’’ 3’’
4’’ 5''
6''
1’’’ 2’’’ 3’’’ 4’’’
5’’’
1’’’’ 2’’’’ 3’’’’ 4’’’’ 5’’’’
6’’’’
O
O OH
OH
O
OH O
O O
H O H
OH Ha Hb
Ha O
O
H OH
O O
O O H
C
H3
Hb
α β
1 4
2 3 5
6
7 8 9
10
1' 2'
3' 4'
5' 6' 1’’
2’’ 3’’
4’’ 5''
6''
1’’’ 2’’’ 3’’’ 4’’’
5’’’
1’’’’ 2’’’’ 3’’’’ 4’’’’ 5’’’’
6’’’’
O
O OH
OH
O
OH O
O O
H O H
OH Ha Hb
Ha O
O
H OH
O O
O O H
C
H3
1135
Rinaldoet al.
Vol. 18, No. 6, 2007
correlation between the hydrogen signal at G 4.06 (H-5''' apiose) and the carbon signal at G 166.3 (E-feruloyl, C=O) thus evidencing the esterification at this position. The foregoing evidences in combination with the downfield shift of the C-5''' apiofuranosyl (G 66.6) when compared to a non-acylated analogue (G 62.4) also supported this conclusion. Therefore, compound (1) was identified as luteolin-7-O-[2''-O-(5'''-O-feruloyl)-β-D-apiofuranosyl]-β -D-glucopyranoside (Figure 1).
A few reports were observed describing the presence of flavonoids in the Nyctaginaceae family. Until now only flavonols were related, such as kaempferol and quercetin from the Bougainvillea glabra6 and B. spectabillis.7 Being thus,Neea theifera is the unique species of Nyctaginaceae, which produces flavones. These data are important because reveal to be able in the future to establish a taxonomic marker in the genus or in this species.
Acknowledgments
We thank Dr. Nivaldo Boralle from Instituto de Química UNESP de Araraquara-SP for recording the NMR spectra, to Dr. Norberto Peporine Lopes from Faculdade de Filosofia Ciências e Letras USP de Ribeirão Preto-SP for HRESIMS analysis, to Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) for funding and fellowships to D. Rinaldo and M. Sannomiya, to Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for grants to W. Vilegas.
Supplementary Information
Supplementary data are available free of charge at http://jbcs.sbq.org.br, as PDF file.
References
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2. Edeoga, H. O.; Ikem, C. I.; S. Afr. J. Bot. 2002,68, 382. 3. Ahmed, B.; Yu, C. P.; Phytochemistry1992,31, 4382. 4. Lami, N.; Kadota, S.; Kikuchi, T.; Chem. Pharm. Bull.1991,
39, 1863.
5. Lami, N.; Kadota, S.; Kikuchi, T.; Momose, Y.;Chem. Pharm. Bull.1991,39, 1551.
6. Heuer, S.; Richter, S.; Metzger, J. W.; Wray, V.; Nimtz, M.; Strack, D.; Phytochemistry1994,37, 761.
7. Chang, W. S.; Lee, Y. J.; Lu, F. J.; Chiang, H. C.; Anti-Cancer Res.1993,13, 2165.
8. Chang, W. S.; Chang, Y. H.; Lu, F. J.; Chiang, H.; C. Anti-Cancer Res.1994,14, 501.
9. De Feo, V.; Piacente, S.; Pizza, C.; Soria, R. U.; Biochem. Syst. Ecol.1998,26, 251.
10. Lavaud, C.; Beauviere, S.; Massiot, G.; Lemenolivier, L.; Bourdy, G.; Phytochemistry1996,43, 189.
11. Duke, J.; Vasquez, R.; Amazonian Ethnobotanical Dictionary, CRC Press: Boca Raton, 1994.
12. Mabry, T. J.; Markham, K. R.; Thomas, M. B.; The Systematic Identification of Flavonoids, Springer: New York, 1970. 13. Harborne, J. B.; The Flavonoids: Advances in Research Since
1986, Chapman and Hall: London, 1996.
14. Agrawal, P. K.;Carbon 13 of Flavonoids, Elsevier: New York, 1989.
15. dos Santos, L.C.; Piacente, S.; Pizza, C.; Toro, R.; Sano, P. T.; Vilegas, W.; Biochem. Syst. Ecol.2002,30, 451.
16. Angyal, S. J.; Bodkin, C. L.; Mills, J. A.; Pojer, P. M.; Aust. J. Chem. 1977,30, 1259.
17. Tronchet, J. M. J.; Tronchet, J.; Carbohydr. Res.1974,34, 263. 18. Kitagawa, I.; Sakagami, M.; Hashiuchi, F.; Zhou, J. L.; Yoshikawa, M.; Ren, J.; Chem. Pharm. Bull.1989,37, 551. 19. Jung, M. J.; Kang, S. S.; Jung, Y. J.; Choi, J. S.; Chem. Pharm.
Bull.2004,52, 1501.
20. Mathias, L.; Vieira, I. J. C.; Braz-Filho, R.; Rodrigues-Filho, E. A.; J. Nat. Prod.1998,61, 1158.
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Received: December 20, 2006 Web Release Date: September 4, 2007
J. Braz. Chem. Soc., Vol. 18, No. 6, S1-S17, 2007. Printed in Brazil - ©2007 Sociedade Brasileira de Química 0103 - 5053 $6.00+0.00
Supplementary InformationSupplementary InformationSupplementary InformationSupplementary InformationSupplementary Information
*e-mail: [email protected]
New Flavone from the Leaves of
Neea theifera
(Nyctaginaceae)
Daniel Rinaldo, Clenilson M. Rodrigues, Juliana Rodrigues, Miriam Sannomiya,
Lourdes C. dos Santos and Wagner Vilegas*
Instituto de Química, Universidade Estadual Paulista, CP 355, 14801-900 Araraquara-SP, Brazil
Figure S1. UV spectrum of (1) (MeOH). 1.00
Wavelength / nm
Intensity
/
au
1.25
400 0.75
250 0.25
0.50
200
206.20 249.49
334.19
300 350
Figure S2. IR spectrum of (1) (KBr disk).
29
25.
84
343
3.0
9
4000 3000 2000 1000
594.
04
819
.70
1031.
86
10
74.
29
1122
.50
1
176.
51
12
59.
44
1332
.73
1
380.
95
145
2.3
1
1514.
03
1
604.
68
165
4.8
3
1
683.
76
170
3.0
4
23
72.
30
29
25.
84
343
3.0
9
29
25.
84
343
3.0
9
4000 3000 2000 1000
Wavenumber / cm-1
T
ransmittance
/
%
0 10 20 30 40 50 60 70 80 90 100
594.
04
819
.70
1031.
86
10
74.
29
1122
.50
1
176.
51
12
59.
44
1332
.73
1
380.
95
145
2.3
1
1514.
03
1
604.
68
165
4.8
3
1
683.
76
170
3.0
4
23
72.
30
29
25.
84
343
3.0
S2 New Flavone from the Leaves of Neea theifera (Nyctaginaceae) J. Braz. Chem. Soc.
Figure S3.1H NMR spectrum of (1) (DMSO-d
6, 11.7 T, TMS, ppm).
5.5 5.0 4.5 4.0 3.5
6.17 6.10 1.65 1.60 3. 7 3 6 3. 7 5 1 3. 76 1 3. 78 0 4. 0 22 4. 04 1 4. 0 5 6 5. 19 0 5. 20 5 5. 37 6
7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1
4.01 3.65
3.651.70 1.77 1.79 1.83 1.71
6. 15 5 6. 1 8 7 6. 3 53 6. 3 5 8 6. 54 5 6. 67 2 6. 6 8 1 6. 6 85 6. 6 8 8 6. 8 4 8 6. 85 1 6. 86 1 6. 87 7 7. 0 6 4 7. 06 7 7. 28 2 7. 3 1 4 7. 33 4 7. 35 1 6. 8 6 8
5.5 5.0 4.5 4.0 3.5
6.17 6.10 1.65 1.60 3. 7 3 6 3. 7 5 1 3. 76 1 3. 78 0 4. 0 22 4. 04 1 4. 0 5 6 5. 19 0 5. 20 5 5. 37 6
5.5 5.0 4.5 4.0 3.5
6.17 6.10 1.65 1.60 3. 7 3 6 3. 7 5 1 3. 76 1 3. 78 0 4. 0 22 4. 04 1 4. 0 5 6 5. 19 0 5. 20 5 5. 37 6
7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1
4.01 3.65
3.651.70 1.77 1.79 1.83 1.71
6. 15 5 6. 1 8 7 6. 3 53 6. 3 5 8 6. 54 5 6. 67 2 6. 6 8 1 6. 6 85 6. 6 8 8 6. 8 4 8 6. 85 1 6. 86 1 6. 87 7 7. 0 6 4 7. 06 7 7. 28 2 7. 3 1 4 7. 33 4 7. 35 1 6. 8 6 8
7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1
4.01 3.65
3.651.70 1.77 1.79 1.83 1.71
6. 15 5 6. 1 8 7 6. 3 53 6. 3 5 8 6. 54 5 6. 67 2 6. 6 8 1 6. 6 85 6. 6 8 8 6. 8 4 8 6. 85 1 6. 86 1 6. 87 7 7. 0 6 4 7. 06 7 7. 28 2 7. 3 1 4 7. 33 4 7. 35 1 6. 8 6 8
δ/ ppm
δ/ ppm
5.5 5.0 4.5 4.0 3.5
6.17 6.10 1.65 1.60 3. 7 3 6 3. 7 5 1 3. 76 1 3. 78 0 4. 0 22 4. 04 1 4. 0 5 6 5. 19 0 5. 20 5 5. 37 6
7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1
4.01 3.65
3.651.70 1.77 1.79 1.83 1.71
6. 15 5 6. 1 8 7 6. 3 53 6. 3 5 8 6. 54 5 6. 67 2 6. 6 8 1 6. 6 85 6. 6 8 8 6. 8 4 8 6. 85 1 6. 86 1 6. 87 7 7. 0 6 4 7. 06 7 7. 28 2 7. 3 1 4 7. 33 4 7. 35 1 6. 8 6 8
5.5 5.0 4.5 4.0 3.5
6.17 6.10 1.65 1.60 3. 7 3 6 3. 7 5 1 3. 76 1 3. 78 0 4. 0 22 4. 04 1 4. 0 5 6 5. 19 0 5. 20 5 5. 37 6
5.5 5.0 4.5 4.0 3.5
6.17 6.10 1.65 1.60 3. 7 3 6 3. 7 5 1 3. 76 1 3. 78 0 4. 0 22 4. 04 1 4. 0 5 6 5. 19 0 5. 20 5 5. 37 6
7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1
4.01 3.65
3.651.70 1.77 1.79 1.83 1.71
6. 15 5 6. 1 8 7 6. 3 53 6. 3 5 8 6. 54 5 6. 67 2 6. 6 8 1 6. 6 85 6. 6 8 8 6. 8 4 8 6. 85 1 6. 86 1 6. 87 7 7. 0 6 4 7. 06 7 7. 28 2 7. 3 1 4 7. 33 4 7. 35 1 6. 8 6 8
7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1
4.01 3.65
3.651.70 1.77 1.79 1.83 1.71
6. 15 5 6. 1 8 7 6. 3 53 6. 3 5 8 6. 54 5 6. 67 2 6. 6 8 1 6. 6 85 6. 6 8 8 6. 8 4 8 6. 85 1 6. 86 1 6. 87 7 7. 0 6 4 7. 06 7 7. 28 2 7. 3 1 4 7. 33 4 7. 35 1 6. 8 6 8
δ/ ppm
δ/ ppm
14 13 12 11 10 9 8 7 6 5 4 3 2 1 0
6.17 4.01
3.65 1.71 1.65 1.70 DMSO Água 2. 4 99 3.44 8 3.73 6 TMS 0.00 12.9 13
14 13 12 11 10 9 8 7 6 5 4 3 2 1 0
6.17 4.01
3.65 1.71 1.65 1.70 DMSO Água 2. 4 99 3.44 8 3.73 6 TMS 0.00 12.9 13
14 13 12 11 10 9 8 7 6 5 4 3 2 1 0
6.17 4.01
3.65 1.71 1.65 1.70 DMSO Água 2. 4 99 3.44 8 3.73 6 TMS 0.00 12.9 13
14 13 12 11 10 9 8 7 6 5 4 3 2 1 0
6.17 4.01
3.65 1.71 1.65 1.70 DMSO Água 2. 4 99 3.44 8 3.73 6 TMS 0.00 12.9 13
δ/ ppm
S3
Rinaldoet al.
Vol. 18, No. 6, 2007
Figure S4.1H NMR 1D-NOESY of (1) (DMSO-d
6, 11.7 T, ppm).
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
7
.067 3.736
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
7
.067 3.736
δ/ ppm
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
7
.067 3.736
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
7
.067 3.736
S4
New Flavone from the Leaves of
Neea theifera
(Nyctaginaceae)
J. Braz. Chem. Soc.
Figur
e S5.
13
C NMR spectrum of (
1
)
(DMSO-d
6, 1
S5
Rinaldo
et al.
V
ol. 18, No. 6, 2007
Figur
e S6.
T
O
CSY
spectrum of (
1
)
(DMSO-d
6, 1
1.7
T
, ppm): a)
Apiose; b) Glucose.
5.0
4.5
4.0
3.5
3.180 3.198
3.215
3.439 3.453 3.477
3.508 3.525 3.539
3.558 3.686 3.706
5.1905.205
3.489
3.468
6.0
5.5
5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
3.751
5.376
b)
a)
5.0
4.5
4.0
3.5
3.180 3.198
3.215
3.439 3.453 3.477
3.508 3.525 3.539
3.558 3.686 3.706
5.1905.205
3.489
3.468
6.0
5.5
5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
3.751
5.376
5.0
4.5
4.0
3.5
3.180 3.198
3.215
3.439 3.453 3.477
3.508 3.525 3.539
3.558 3.686 3.706
5.1905.205
3.489
3.468
5.0
4.5
4.0
3.5
3.180 3.198
3.215
3.439 3.453 3.477
3.508 3.525 3.539
3.558 3.686 3.706
5.1905.205
3.489
3.468
6.0
5.5
5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
3.751
5.376
6.0
5.5
5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
3.751
5.376
b)
a)
δ
/
ppm
δ
/
ppm
5.0
4.5
4.0
3.5
3.180 3.198
3.215
3.439 3.453 3.477
3.508 3.525 3.539
3.558 3.686 3.706
5.1905.205
3.489
3.468
6.0
5.5
5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
3.751
5.376
b)
a)
5.0
4.5
4.0
3.5
3.180 3.198
3.215
3.439 3.453 3.477
3.508 3.525 3.539
3.558 3.686 3.706
5.1905.205
3.489
3.468
6.0
5.5
5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
3.751
5.376
5.0
4.5
4.0
3.5
3.180 3.198
3.215
3.439 3.453 3.477
3.508 3.525 3.539
3.558 3.686 3.706
5.1905.205
3.489
3.468
5.0
4.5
4.0
3.5
3.180 3.198
3.215
3.439 3.453 3.477
3.508 3.525 3.539
3.558 3.686 3.706
5.1905.205
3.489
3.468
6.0
5.5
5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
3.751
5.376
6.0
5.5
5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
3.751
5.376
b)
a)
δ
/
ppm
δ
/
S6 New Flavone from the Leaves of Neea theifera (Nyctaginaceae) J. Braz. Chem. Soc.
δ/ ppm (F1)
180 160 140 120 100 80.0 60.0 40.0 20.0
δ
/p
pm
(F
2
)
13.0 12.0 11.0 10.0 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 -1.0
200
δ/ ppm (F1)
180 160 140 120 100 80.0 60.0 40.0 20.0
δ
/p
pm
(F
2
)
13.0 12.0 11.0 10.0 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 -1.0
200
Figure S7.gHMQC spectrum of (1) (DMSO-d6, 11.7 T, TMS, ppm).
Figure S8.gHMBC spectrum of (1) (DMSO-d6, 11.7 T, TMS, ppm).
200 180 160 140 120 100 80.0 60.0 40.0 20.0
13.0 12.0 11.0 10.0 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 -1.0
δ/ ppm (F1)
200 180 160 140 120 100 80.0 60.0 40.0 20.0
δ
/p
pm
(F
2
)
13.0 12.0 11.0 10.0 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 -1.0
200 180 160 140 120 100 80.0 60.0 40.0 20.0
13.0 12.0 11.0 10.0 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 -1.0
δ/ ppm (F1)
200 180 160 140 120 100 80.0 60.0 40.0 20.0
δ
/p
pm
(F
2
)
S7
Rinaldoet al.
Vol. 18, No. 6, 2007
Figure S9. ESI-MS spectrum of (1) (negative mode, 70 V).
755 285
284
283
561
286 299
447
365 448 543
579
562
563 580
581595
753 756
836 757
835 771
801820
837
200 250 300 350 400 450 500 550 600 650 700 750 800 850
m/z 0
100
A
b
und
anc
e
/
%
365
595 771
Figure S10.1H NMR spectrum of vitexin (DMSO-d
6, 4.7 T, TMS, ppm).
14 13 12 11 10 9 8 7 6 5 4 3 2 1 0
2.25
2.20 1.01 1.09
1.00
DMSO
4.
7
0
4.75
6.21
6.71
6.90
7.9
6
8.
0
0
13
.1
3
6.86
TMS
8.0 7.5 7.0 6.5 6.0 5.5 5.0
2.25
2.20 1.02 1.01 1.09
4.70
4.
75
6.
21
6.7
1
6.
86
6.90
7.96
8.
00
2.49
0.00
14 13 12 11 10 9 8 7 6 5 4 3 2 1 0
2.25
2.20 1.01 1.09
1.00
DMSO
4.
7
0
4.75
6.21
6.71
6.90
7.9
6
8.
0
0
13
.1
3
6.86
TMS
8.0 7.5 7.0 6.5 6.0 5.5 5.0
2.25
2.20 1.02 1.01 1.09
4.70
4.
75
6.
21
6.7
1
6.
86
6.90
7.96
8.
00
2.49
S8
New Flavone from the Leaves of
Neea theifera
(Nyctaginaceae)
J. Braz. Chem. Soc.
Figur
e S1
1.
13
C NMR spectrum of vitexin (DMSO-d6, 4.7
T
, ppm).
200
150
100
50
0
DMSO
39.50 61.30
70.5570.91 73.46
78.71 81.82 98.35102.44
104.64
115.89 121.62 128.93 156.01 160.44161.23
163.85 182.01
180
170
160
150
140
130
120
110
100
90
80
70
60 61.30
70.5570.91 73.46
78.71 81.82 98.35
102.44 104.64
115.89 121.62 128.93 156.01
160.44161.23 163.85
182.01
163.29
200
150
100
50
0
DMSO
39.50 61.30
70.5570.91 73.46
78.71 81.82 98.35102.44
104.64
115.89 121.62 128.93 156.01 160.44161.23
163.85 182.01
180
170
160
150
140
130
120
110
100
90
80
70
60 61.30
70.5570.91 73.46
78.71 81.82 98.35
102.44 104.64
115.89 121.62 128.93 156.01
160.44161.23 163.85
182.01
163.29
Figur
e S12.
ESI-MS spectrum of vitexin (negative mode, 70
S9
Rinaldoet al.
Vol. 18, No. 6, 2007
Figure S13.1H NMR spectrum of the mixture of isovitexin and vitexin (DMSO-d
6, 4.7 T, TMS, ppm).
15 14 13 12 11 10 9 8 7 6 5 4 3 2 1 0
6.05
4.08 2.05
2.02 1.04
1.00
TMS DMSO
0.
0
0
2.
49
4.5
7
4.
6
2
4.7
0
4.
7
5
6.
21
6.41
6.
71
6.
8
6
6.
90
7.
86
7.
90
7.
96
8.0
0
13
.1
2
13
.5
6
2.08
8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5
6.05
4.082.04 3.04 1.04 2.051.01
4.
57
4.
62
4.
70
4.
75
6.
21
6.
41
6.
71
6.
86
6.
90
7.
86
7.
90
7.
96
8.
00
2.08
15 14 13 12 11 10 9 8 7 6 5 4 3 2 1 0
6.05
4.08 2.05
2.02 1.04
1.00
TMS DMSO
0.
0
0
2.
49
4.5
7
4.
6
2
4.7
0
4.
7
5
6.
21
6.41
6.
71
6.
8
6
6.
90
7.
86
7.
90
7.
96
8.0
0
13
.1
2
13
.5
6
2.08
15 14 13 12 11 10 9 8 7 6 5 4 3 2 1 0
6.05
4.08 2.05
2.02 1.04
1.00
TMS DMSO
0.
0
0
2.
49
4.5
7
4.
6
2
4.7
0
4.
7
5
6.
21
6.41
6.
71
6.
8
6
6.
90
7.
86
7.
90
7.
96
8.0
0
13
.1
2
13
.5
6
2.08
8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5
6.05
4.082.04 3.04 1.04 2.051.01
4.
57
4.
62
4.
70
4.
75
6.
21
6.
41
6.
71
6.
86
6.
90
7.
86
7.
90
7.
96
8.
00
2.08
S10
New Flavone from the Leaves of
Neea theifera
(Nyctaginaceae)
J. Braz. Chem. Soc.
Figur
e S15.
1
H NMR spectrum of isoorientin
(DMSO-d
6,
4.7 T
, TMS,
ppm).
15
14
13
12
11
10
9
8
7
6
5
4
3
2
1
2.10
1.34
1.25
1.00
DMSO
Água
2.49
3.45 4.54
4.59 6.47 6.66 6.89
7.38 7.42 13.57
1.05
0 TMS
0.00
7.5
7.0
6.5
6.0
5.5
5.0
4.5
2.10
1.34
1.25
1.05
1.03
4.54 4.59
6.47 6.66 6.85
6.89
7.38 7.42
15
14
13
12
11
10
9
8
7
6
5
4
3
2
1
2.10
1.34
1.25
1.00
DMSO
Água
2.49
3.45 4.54
4.59 6.47 6.66 6.89
7.38 7.42 13.57
1.05
0 TMS
0.00
7.5
7.0
6.5
6.0
5.5
5.0
4.5
2.10
1.34
1.25
1.05
1.03
4.54 4.59
6.47 6.66 6.85
6.89
7.38 7.42
Figur
e S16.
13
C NMR spectrum of isoorientin
(DMSO-d
6, 4.7
T
, ppm).
200
150
100
50
0
DMSO
39.50 60.48
70.3870.81 73.2378.48
81.76 93.79 103.00103.59
109.03 113.43 119.26121.62 146.01149.98
156.51160.90 163.95 182.18
180
170
160
150
140
130
120
110
100
90
80
70
60
60.48 70.3870.81
73.23 78.48 81.76 93.79 103.00103.59
109.03 113.43 116.31
119.26121.62 146.01
149.98 156.51
160.90 163.95 182.18
200
150
100
50
0
DMSO
39.50 60.48
70.3870.81 73.2378.48
81.76 93.79 103.00103.59
109.03 113.43 119.26121.62 146.01149.98
156.51160.90 163.95 182.18
180
170
160
150
140
130
120
110
100
90
80
70
60
60.48 70.3870.81
73.23 78.48 81.76 93.79 103.00103.59
109.03 113.43 116.31
119.26121.62 146.01
149.98 156.51
S11
Rinaldoet al.
Vol. 18, No. 6, 2007
Figure S17. ESI-MS spectrum of isoorientin (negative mode, 70 V).
Figure S18.1H NMR spectrum of orientin (DMSO-d
6, 4.7 T, TMS, ppm).
4.7 4.6
1.24
4.
663
4.
682
13 12 11 10 9 8 7 6 5 4 3 2 1 0
1.24 1.05
1.00 0.94 0.80
TMS DMSO
Água
0.
00
0
2.
48
7
3.
33
3
4.
66
3
4.
68
2
6.
25
4
6.
61
9
6.
86
1
7.
46
3
7.
50
7
7.
52
4
13.
143
7.5 7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1
1.05
0.94 0.79 0.82 0.80
6.
2
54
6.
619
6.
845
6.
861
7.
463
7.
50
7
7.
524
4.7 4.6
1.24
4.
663
4.
682
4.7 4.6
1.24
4.
663
4.
682
4.7 4.6
1.24
4.
663
4.
682
13 12 11 10 9 8 7 6 5 4 3 2 1 0
1.24 1.05
1.00 0.94 0.80
TMS DMSO
Água
0.
00
0
2.
48
7
3.
33
3
4.
66
3
4.
68
2
6.
25
4
6.
61
9
6.
86
1
7.
46
3
7.
50
7
7.
52
4
13.
143
13 12 11 10 9 8 7 6 5 4 3 2 1 0
1.24 1.05
1.00 0.94 0.80
TMS DMSO
Água
0.
00
0
2.
48
7
3.
33
3
4.
66
3
4.
68
2
6.
25
4
6.
61
9
6.
86
1
7.
46
3
7.
50
7
7.
52
4
13.
143
7.5 7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1
1.05
0.94 0.79 0.82 0.80
6.
2
54
6.
619
6.
845
6.
861
7.
463
7.
50
7
7.
524
7.5 7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1
1.05
0.94 0.79 0.82 0.80
6.
2
54
6.
619
6.
845
6.
861
7.
463
7.
50
7
7.
S12 New Flavone from the Leaves of Neea theifera (Nyctaginaceae) J. Braz. Chem. Soc.
Figure S19.13C NMR spectrum of orientin (DMSO-d
6, 4.7 T, ppm).
4.7 4.6
1.24
4.
663
4.
682
13 12 11 10 9 8 7 6 5 4 3 2 1 0
1.24 1.05
1.00 0.94 0.80
TMS DMSO
Água
0.
00
0
2.
48
7
3.
33
3
4.
66
3
4.
68
2
6.
25
4
6.
61
9
6.
86
1
7.
46
3
7.
50
7
7.
52
4
13.
143
7.5 7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1
1.05
0.94 0.79 0.82 0.80
6.
2
54
6.
619
6.
845
6.
861
7.
463
7.
50
7
7.
524
4.7 4.6
1.24
4.
663
4.
682
4.7 4.6
1.24
4.
663
4.
682
4.7 4.6
1.24
4.
663
4.
682
13 12 11 10 9 8 7 6 5 4 3 2 1 0
1.24 1.05
1.00 0.94 0.80
TMS DMSO
Água
0.
00
0
2.
48
7
3.
33
3
4.
66
3
4.
68
2
6.
25
4
6.
61
9
6.
86
1
7.
46
3
7.
50
7
7.
52
4
13.
143
13 12 11 10 9 8 7 6 5 4 3 2 1 0
1.24 1.05
1.00 0.94 0.80
TMS DMSO
Água
0.
00
0
2.
48
7
3.
33
3
4.
66
3
4.
68
2
6.
25
4
6.
61
9
6.
86
1
7.
46
3
7.
50
7
7.
52
4
13.
143
7.5 7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1
1.05
0.94 0.79 0.82 0.80
6.
2
54
6.
619
6.
845
6.
861
7.
463
7.
50
7
7.
524
7.5 7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1
1.05
0.94 0.79 0.82 0.80
6.
2
54
6.
619
6.
845
6.
861
7.
463
7.
50
7
7.
524
S13
Rinaldo
et al.
V
ol. 18, No. 6, 2007
Figur
e S21.
1
H NMR spectrum of vicenin-2
(DMSO-d
6,
11.7 T
, TMS,
ppm).
14
13
12
11
10
9
8
7
6
5
4
3
2
1
0
2.08
2.27
2.25
1.00
TMS
DMSO
Á
gua
0.000
2.503 3.411 4.755
4.7754.799 4.819
6.793 6.913 8.0148.030 13.735
8.0
7.5
7.0
6.5
6.0
5.5
5.0
2.08
2.27
2.25
1.08
4.7554.775
4.799 4.819
6.793
6.8976.913
8.0148.030
14
13
12
11
10
9
8
7
6
5
4
3
2
1
0
2.08
2.27
2.25
1.00
TMS
DMSO
Á
gua
0.000
2.503 3.411 4.755
4.7754.799 4.819
6.793 6.913 8.0148.030 13.735
8.0
7.5
7.0
6.5
6.0
5.5
5.0
2.08
2.27
2.25
1.08
4.7554.775
4.799 4.819
6.793
6.8976.913
8.0148.030
Figur
e S22.
13
C NMR spectrum of vicenin-2
(DMSO-d
6, 1
1.7
T
, TMS, ppm).
200
150
100
50
0
DMSO
-d
6
39.50
59.86 60.28
61.33 69.04
70.61 71.96
72.28 77.89
78.84 80.91 81.96
102.61 105.30
107.59 115.89
121.53 129.03 155.13
158.67 161.26
164.05 182.31
180
170
160
150
140
130
120
110
100
90
80
70
60
59.86 60.28
69.04 70.61 72.28 73.43
77.8978.84
80.91 81.96
102.61 105.30
107.59
115.89 121.53
129.03 155.13
158.67
161.26 164.05
182.31
74.15 103.66
200
150
100
50
0
DMSO
-d
6
39.50
59.86 60.28
61.33 69.04
70.61 71.96
72.28 77.89
78.84 80.91 81.96
102.61 105.30
107.59 115.89
121.53 129.03 155.13
158.67 161.26
164.05 182.31
200
150
100
50
0
DMSO
-d
6
39.50
59.86 60.28
61.33 69.04
70.61 71.96
72.28 77.89
78.84 80.91 81.96
102.61 105.30
107.59 115.89
121.53 129.03 155.13
158.67 161.26
164.05 182.31
180
170
160
150
140
130
120
110
100
90
80
70
60
59.86 60.28
69.04 70.61 72.28 73.43
77.8978.84
80.91 81.96
102.61 105.30
107.59
115.89 121.53
129.03 155.13
158.67
161.26 164.05
182.31
S14 New Flavone from the Leaves of Neea theifera (Nyctaginaceae) J. Braz. Chem. Soc.
Figure S23. TOCSY spectrum of vicenin-2 (DMSO-d6, 11.7 T, ppm, irradiation of G4.809).
5.0 4.5 4.0 3.5 3.0
3.2
76
3.
29
5
3.3
19
3.
3
39
3.3
67
3.
3
86
3.
4
05
3.4
22
3
.508
3.5
15
3.5
33
3.6
46
3.7
59
3.7
77
3.8
70
3.
88
8
3.
90
6
4.7
55
4
.775
4
.799
4
.819
5.0 4.5 4.0 3.5 3.0
3.2
76
3.
29
5
3.3
19
3.
3
39
3.3
67
3.
3
86
3.
4
05
3.4
22
3
.508
3.5
15
3.5
33
3.6
46
3.7
59
3.7
77
3.8
70
3.
88
8
3.
90
6
4.7
55
4
.775
4
.799
4
.819
S15
Rinaldoet al.
Vol. 18, No. 6, 2007
Figure S25.1H NMR spectrum of chrysoeriol (DMSO-d
6, 11.7 T, ppm).
8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
3.22 2.25 1.09 0.90 0.96
DMSO Água
2.
50
0
3.
3
58
3.89
0
6.
1
89
6.
19
3
6.5
01
6.87
5
6.
9
28
6.94
6
7.
5
49
7.56
1
TMS
0.
0
00
6.95 6.80 6.65 6.50 6.35 6.15
1.09 1.00 0.90 0.96
6.
18
9
6.
193
6.
501
6.
505
6.
875
6.
928
6.9
45
7.59 7.50
2.25
7.
548
7.56
1
7.
56
5
8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
3.22 2.25 1.09 0.90 0.96
DMSO Água
2.
50
0
3.
3
58
3.89
0
6.
1
89
6.
19
3
6.5
01
6.87
5
6.
9
28
6.94
6
7.
5
49
7.56
1
TMS
0.
0
00
6.95 6.80 6.65 6.50 6.35 6.15
1.09 1.00 0.90 0.96
6.
18
9
6.
193
6.
501
6.
505
6.
875
6.
928
6.9
45
7.59 7.50
2.25
7.
548
7.56
1
7.
56
5
Figure S26. UV spectrum of chrysoeriol (MeOH). 50
100 150
200 250 300 350 nm
206
.4
5
25
0.
50
3
46.
19
50 100 150
mAU
200 250 300 350
wavelenght / nm
206
.4
5
25
0.
50
3
46.
S16 New Flavone from the Leaves of Neea theifera (Nyctaginaceae) J. Braz. Chem. Soc.
Figure S27.1H NMR 1D-NOESY of chrysoeriol (DMSO-d
6, 11.7 T, ppm, irradiation of G3.890).
9 8 7 6 5 4 3 2 1 0
3.
8
90
7
.548
9 8 7 6 5 4 3 2 1 0
3.
8
90
7
.548
S17
Rinaldoet al.
Vol. 18, No. 6, 2007
Figure S29. Cromatograms of the co-injections of a) apigenin isolated + apgenin Sigma® (1:1 m/m) and b) luteolin isolated + luteolin Sigma® (1:1 m/m)
(C18, 250 × 4.60 mm i. d. × 5 µm). A binary gradient elution system with solvent A (0.05% TFA in H2O) and solvent B (0.05% TFA in CH3CN) was used,
with linear gradient starting from 68:32 (A:B) at 20 min and then changed to 75:35 (A:B) for 40 min. The flow-rate was 1.0 mL min-1.λ 254 nm.
10 20 30 40 50 60
0.0 0.5 1.0 1.5
0 50 100 150 200 250
100 200 300 400
200 250 300 350 nm
20
9
.13 26
6.46
33
5.89
100 200 300 400 500 600 700
200 250 300 350 nm
207.86
2
52.
78
3
47.18
10 20 30 40 50 60
0.0 0.5 1.0 1.5
0 50 100 150 200 250
100 200 300 400
200 250 300 350 nm
20
9
.13 26
6.46
33
5.89
100 200 300 400 500 600 700
200 250 300 350 nm
207.86
2
52.
78
3
47.18
10 20 30 40 50 60
time / min 0.0
0.5 1.0 1.5
AU
0 50 100 150 200 250
mAU
100 200 300 400
200 250 300 350 nm
20
9
.13 26
6.46
33
5.89
100 200 300 400
mAU
200 250 300 350 nm
20
9
.13 26
6.46
33
5.89
100 200 300 400 500 600 700
200 250 300 350 nm
207.86
2
52.
78
3
47.18
100 200 300 400 500 600 700
mAU
200 250 300 350 nm
207.86
2
52.
78
3
47.18
a)
