Supporting Material Antioxidant Flavonoids from Asteriscus maritimus (L.) Less. Habiba Daroui-Mokaddem a,b, Ahmed Kabouche a, Naima Boutaghane a, Claude Alain Calliste c, Jean-Luc Duroux c, Zahia Kabouche a,* a
Université des frères Mentouri-Constantine, Département de chimie, Laboratoire
d’Obtention de Substances Thérapeutiques (LOST), Campus Chaabet Ersas, 25000 Constantine, Algeria b
Département de Médecine, Faculté de Médecine, Université Badji-Mokhtar-Annaba,
23000 Annaba, Algeria c
Laboratoire de Biophysique, UPRES EA-1085, Biomolécules et cibles cellulaires
tumorales Faculté de Pharmacie, 2 rue du Dr. Marcland, 87025 Limoges Cedex, France
[email protected] (Z. KABOUCHE)
One new flavonol glycoside, patuletin-7-O-[6-O-caffeoyl-2-O-[(S)-3hydroxyisobutanoyl)]glucopyranoside] (Astermaritimoside) (1) and four known flavonoids, patuletin-7-O-β-[-6-O-[(S)-3-hydroxyisobutanoyl)glucopyranoside] (2) patuletin-7-O-[6-O-caffeoylglucopyranoside] (3), patuletin-7-O-β-D-glucopyranoside (4) and quercetin-3-O-β-rutinoside (5) have been isolated from Asteriscus maritimus (L.) Less. Chemical structures were elucidated by mass spectrometry, 1H NMR and 13 C NMR, COSY, HMQC and HMBC techniques. The antioxidative effect was evaluated by the Electronic Spin Resonance (ESR) method in order to visualize the inhibition of the DPPH radical. The n-butanol fraction was found to possess a remarkable antioxidant activity that was correlated with the total amount of phenolics. The isolated compounds exhibited a good antioxidant activity. Key words: Asteriscus maritimus (L.) Less., Asteraceae, flavonoid, antioxidant activity, ESR.
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Table of Contents
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Figure S1
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MS and NMR Supplementary data S1. HRESI-MS Spectrum of Compound 1
3
S2. 1H-NMR (400 MHz, CD3OD) Spectrum of Compound 1
4
S3. Expansion of the 1H-NMR (400 MHz) Spectrum of Compound 1
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S4. 13C RMN (100 MHz, CD3OD) Spectrum of Compound 1
5
S5. Expansion of the 13C RMN (100 MHz) Spectrum of compound 1
5
S6. COSY (400 MHz) Spectrum of Compound 1
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S7. Expansion of the COSY (400 MHz) Spectrum of Compound 1
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S8. Expansion of the HMQC (400 MHz) Spectrum of compound 1
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S9. HMBC (400 MHz) Spectrum of compound 1
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S10. HMBC (400 MHz) Spectrum of Compound 1
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S11. Expansion of the HMBC (400 MHz) Spectrum of Compound 1
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S12. Expansion of the HMBC (400 MHz) Spectrum of compound 1
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Antioxidant activity
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1
Figure S1. Structures of compounds (2-5) isolated from Asteriscus maritimus
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Fédération de Recherche Physique et Chimie du Vivant (FR2708 : CBM/ICOA)
HRMS
Plate-forme de Spectrométrie de Masse Haute Résolution Analysis Info Sample Name Analysis Name Method
Acquisition Date 21/06/2012 11:49:56 H2 X004366CYC.d Positif.m
Laboratory Instrument / Ser# maXis
255552.00 086
Acquisition Parameter Source Type Focus Scan Begin Scan End
ESI Not active 50 m/z 3000 m/z
Ion Polarity Set Capillary Set End Plate Offset Set Collision Cell RF
Positive 4500 V -500 V 1000.0 Vpp
Set Nebulizer Set Dry Heater Set Dry Gas Set Divert Valve
Intens. x10
0.6 Bar 200 °C 7.0 l/min Waste
+MS, 0.16-0.93min #9-55, -Peak Bkgrnd
4
1+ 657.1440
4 2
1+ 743.1803 163.0391
0 100 Intens. 1.
321.2767
200
495.1128
300
400
500
600
743.1803
700
800 m/z
+MS, 0.16-0.93min #9-55, -Peak Bkgrnd
4
x10 0.8 1+ 744.1839
0.6 0.4
1+ 745.1863
0.2 0.0
1+ 743.1818
4
x10 1.
1+ 746.1895 C₃₃ H₃₃ O₃₃, 743.1823
1.0 0.8 1+ 744.1852
0.6 0.4
1+ 745.1877
0.2 0.0
743
Meas. m/z
# Ion Formula
657.143962
1 2 1 1 1 2 1 1
679.126096 743.180267 765.162614
C31H29O16 C44H21N2O5 C31H28NaO16 C44H20N2NaO5 C35H35O18 C48H27N2O7 C35H34NaO18 C48H26N2NaO7
744
745
1+ 746.1903 746
m/z
err [ppm]
mSigma
657.145011 657.144498 679.126956 679.126442 743.181791 743.181278 765.163735 765.163222
1.6 0.8 1.3 0.5 2.0 1.4 1.5 0.8
2.4 75.2 8.8 65.0 19.8 47.2 6.0 62.6
747
rdb e¯ Conf 17.5 35.5 17.5 35.5 18.5 36.5 18.5 36.5
S1. HRESI-MS Spectrum of Compound 1
3
even even even even even even even even
748
N-Rule ok ok ok ok ok ok ok ok
m/z
S2. 1H-NMR (400 MHz, CD3OD) Spectrum of Compound 1
S3. 1H-NMR (400 MHz, CD3OD) Spectrum of Compound 1
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S4. 13C RMN (100 MHz, CD3OD) Spectrum of Compound 1
C6’C6 ’
S5. 13C RMN (100 MHz, CD3OD) Spectrum of compound 1
5
S6. COSY (400 MHz) Spectrum of Compound
S7. COSY (400 MHz) Spectrum of Compound 1
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S8. HMQC (400 MHz) Spectrum of compound 1
S9. HMBC (400 MHz) Spectrum of compound 1
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S10. HMBC (400 MHz) Spectrum of Compound 1
S11. HMBC (400 MHz) Spectrum of Compound 1
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S12. HMBC (400 MHz) Spectrum of compound 1
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Antioxidant activity: The potential antioxidant activity of plant extracts was assessed on the basis of the scavenging activity of the stable DPPH free radical. Reaction mixtures contained 100 µL test samples (0.5 mg/mL dissolved in each solvent) and 100 µL of DPPH ethanolic solution (5×10-4m). Due to its paramagnetic properties DPPH exhibits an ESR signal. ESR spectra were obtained with a bruker esp 300e spectrometer using micro-sampling pipets at room temperature under the following conditions: modulation frequency, 100 kHz; microwave frequency, 9.78 GHz; microwave power, 2 mw; modulation amplitude, 1.97 g; time constant, 10.24 ms. All spectra were recorded 3 min after agitation. The inhibition ratio was calculated using:
Inhibition ratio:
(ref - extract) ____________ (ref - bg)
Where ref is the double integral of the reference signal (DPPH plus solvent), extract is the double integral of the test signal (DPPH plus solvent plus plant extract), and bg is the back-ground signal. The data were the average of four measurements.
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