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Universidade Federal Rural do Rio de Janeiro, Departamento de Qumica, Br 465, Km. 07, CEP: 23980-000; Seropedica-RJ,
Brazil
Grupo Metabolomica-LCQUI-CCT,
Universidade Estadual do Norte Fluminense, Av. Alberto Lamego, 2000, CEP: 28015-620, Campos-RJ, Brazil
The 1 H and 13 C NMR spectra of 5-acetyl-7,4 -dimethoxyflavone-(68 )-5 -acetyl-7 ,4 -dimethoxyflavone,
a new agathisflavone derivative, were completely assigned on the basis of 1D and 2D NMR techniques.
Copyright 2005 John Wiley & Sons, Ltd.
INTRODUCTION
In a previous report, the structure determination of a
new flavone dimer isolated from the leaves of Ouratea
hexasperma (Ochnaceae) was described.1 Continuing the
phytochemical and pharmacological investigation of Ouratea
species,2 5 we have isolated a reasonable amount of
700 -methyl-agathisflavone, an abundant constituent from
O. hexasperma, collected both in the Amazon cerrado, Amapa
state,1 and in the Mata Atlanica4 region near Joao Pessoa,
Paraiba state, Brazil. The NMR spectral investigation of the
new agathisflavone derivative permitted the confirmation of
the expected structure, the assignment of the 1 H and 13 C
chemical shifts and the proposal of a conformer to explain
some signal distortion in the 1 H NMR spectra.
EXPERIMENTAL
General procedures
Fourier transform NMR spectra, 13 C (BroadBand Decoupling (BBD) and Attached Proton Test (APT)), 1 H 1 H-COSY,
HMQC, HMBC and NOESY, were performed using standard
pulse sequences from the Delta NMR software (version 4.3)
of the JEOL Eclipse C 400 spectrometer operating at 400 MHz
for 1 H and 100 MHz for 13 C. Typical NMR parameters for
the 1 H NMR were pulse angle of 45 , spectral size of 16 K,
and a spectral width of 15 ppm. Heteronuclear 1 H 13 CCOSY (HMQC and HMBC) experiments were modulated
with 1 JCH D 140 Hz and n JCH (n D 2 and 3 D 9 Hz. The
Nuclear Overhauser Effect (NOE)-difference spectroscopy
experiment was recorded on a Bruker AC 200 spectrometer operating at 200 MHz for 1 H using the Bruker program
Correspondence to: Mario Geraldo de Carvalho, Universidade
Federal Rural do Rio de Janeiro, Departamento de Qumica, Br 465,
Km. 07, CEP: 23980-000; Seropedica-RJ, Brazil.
E-mail: mgeraldo@ufrrj.br
Contract/grant sponsor: CNPq.
Contract/grant sponsor: FAPERJ.
Contract/grant sponsor: CAPES.
36
M. G. de Carvalho et al.
of AA0 BB0 systems with CH 127.8 (C-20 ,60 ), 127.7 (C-2000 ,6000 ),
114.5 (C-30 ,50 ), 114.4 ppm (C-3000 ,5000 ) and of H-3, 300 ,600 and
8 with CH 107.4 (C-3), 106.4 (C-300 ), 103.7 (C-600 ) and
97.0 ppm (C-8), respectively. This spectrum showed signals
of methylic carbons at CH3 56.4 (H3 C O-700 ), 56.3 (H3 CO-7),
55.5 (H3 CO-4000 ), 55.4 (H3 CO-40 ), 20.9 (H3 CCOO-5) and
21.2 ppm (H3 CCOO-500 ), which were attributed to the cross
peaks revealed in the HMQC experiment with H 3.84, 3.82,
3.78, 3.91, 2.15 and 2.49 ppm, respectively. The signals due
to 2 JCH observed in the HMBC experiment of the methyl
group at 2.15 (br s) and 2.49 ppm (s) with CO 168.7 and
169.6 ppm, respectively, confirmed the acetyl groups. The
chemical shifts of the quaternary carbons of the flavone
moieties of 3 were established by 1 H 13 C COSY (HMBC)
spectral analysis. The cross peaks of H-3 with C-2, C-4 (2 JCH ),
C-10, and C-10 (3 JCH ), H-300 with C-200 , C-400 (2 JCH ), C-1000 , and
C-1000 (3 JCH ), H-600 with C-500 , C-700 (2 JCH ), C-1000 , and C-800
(3 JCH ), and H-8 with C-9, C-7 (2 JCH ), C-10, and C-6 (3 JCH ) were
used to attribute the chemical shifts of C-800 with 107.1,
C-1000 with 111.0, C-10 with 111.3, C-6 with 107.1, C-10 ,1000
with 123.7 and C-500 with 150.9, C-9 with 158.8, C-700 with
2,6
2,6
3,5,8
3,5
6
3 3
MeO-4/H-3,5
1
6.0
MeO-7/H-6
Hx1H-COSY
MeO-7/H-8
MeO-4/H-3,5
6
7.0
JH/H
3,5
3/2,6
3/2,6 NOE
8.0
8.0
7.0
6.0
5.0
4.0
3.0
2.0 1
Figure 3. Conformers of 3.
3.82* (H3 CO-7) and 3.78 ppm (H3 CO-4000 ). The 1 Hf1 HgNOE-difference spectra performed with irradiation at H-300
(showed an NOE at the H-2000 ,6000 signal), H-3 (showed NOE
at H-20 ,60 ) and at the methoxyl groups at 3.91 and 3.78 ppm
(showed NOE at H-30 , 50 and 3000 ,5000 , respectively) were used
to confirm those assignments. The assigned signals in the
NOESY expectrum (Fig. 2) confirmed those experiments
and were also used in the 1 H signal assignments. The
broadening of the H-2000 ,6000 , H3 CO-7 and 700 signals assigned
by (*), besides the signal of H3 CCOO-5 (br s, Fig. 2), can
be justified by magnetic anisotropy of the groups according
to the conformer (Fig. 3) because of the restricted rotation
about the bond connecting both the flavone moieties. The
analysis of the 13 C NMR spectra (BBD and APT) was
used to identify the number of bonded hydrogens for each
carbon signal. The heteronuclear correlations via one bond
(1 H 13 C COSY, HMQC) showed cross peaks of hydrogens
HMQC (1 JCH )
13
C/1 H
H (mult,
J in Hz)
CH
3
300
600
8
20 ,60
2000 ,6000
30 ,50
3000 ,5000
H3 CO-40
H3 CO-4000
H3 CO-7
H3 CO-700
H3 CCO2 -5
H3 CCO2 -5
H3 CCO2 -500
H3 CCO2 -500
107.4
106.4
103.7
97.0
127.8b
127.7b
114.5c
114.4c
55.4d
55.5d
56.3
56.4
20.9
168.7
21.2
169.6
HMBC
( 2 JH C
6.61 (s)
6.50 (s)
6.73 (s)
7.03 (s)
7.89 (d, 8,8)
7.45 (br d, 8)e
7.05 (d, 8,8)
6.84 (d, 8,8)
3.91 (s)
3.78 (s)
3.82 (br s)e
3.84 (br s)e
2.15 (br s)e
C-2,C-4
C-200 ,C-400
C-500 ,C-700
C-9,C-7
C-30 , 50
C-3000 ,5000
C-40
C-4000
2.49 (s)
H3 CCO2 -500
(3 JH C
C-10,C-10
C-1000 ,C-1000
C-1000 ,C-800
C-10,C-6
C-40
C-4000
C-10
C-1000
C-40
C-4000
C-7
C-700
H3 CCO2 -5
2
200
4
400
5
500
161.9
162.2
176.5
176.9
155.7
150.9
6
7
700
800
9
900
107.1
161.7
161.0
107.1
158.8
158.8
10
1000
10
1000
40
4000
111.3b
111.0b
123.7
123.7
162.4
162.2
161.0, C-2 with 161.9, C-200 with 162.2, C-4 with 176.5 and
C-400 with 176.9 ppm (Table 1). The cross peaks of H3 CO
with C 161.0 (C-700 ), 161.7 ppm (C-7) and with 162.4 and
162.2 ppm were used to confirm the chemical shifts of the
carbons that support these groups defining the C of 40 and 4000
as 162.4 and 162.2 ppm, respectively. The HMBC spectrum
did not show cross peaks with the signal of C 155.7 ppm,
corroborating that C-5 carries the protected acetyl group.
Acknowledgements
The authors are grateful to CNPq, FAPERJ and CAPES for a research
fellowship and for financial support.
REFERENCES
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