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In vitro synergistic antioxidant activity and identification of antioxidant components from Astragalus membranaceus and Paeonia lactiflora.

Xu X, Li F, Zhang X, Li P, Zhang X, Wu Z, Li D - PLoS ONE (2014)

Bottom Line: Many traditionally used herbs demonstrate significantly better pharmacological effects when used in combination than when used alone.Among them, E1 exhibited the strongest synergistic effect in scavenging DPPH radicals and reducing ferric ions (P<0.05).A strong correlation between the increment of total phenolic/flavonoid and synergistic antioxidant activity, especially between the increment of total flavonoid and the increase in ferric reducing power was observed.

View Article: PubMed Central - PubMed

Affiliation: Department of Food Science, Shandong Agricultural University, Taian, China.

ABSTRACT
Many traditionally used herbs demonstrate significantly better pharmacological effects when used in combination than when used alone. However, the mechanism underlying this synergism is still poorly understood. This study aimed to investigate the synergistic antioxidant activity of Astragalus membranaceus (AME) and Paeonia Lactiflora (PL), and identify the potential antioxidant components by 1,1-diphenyl-2-picrylhydrazine (DPPH) radical spiking test followed by a high performance liquid chromatography separation combined with diode array detection and tandem mass spectrometry analysis (DPPH-HPLC-DAD-MS/MS). Eight AME-PL combined extracts (E1-E8) were prepared based on bioactivity-guided fractionation. Among them, E1 exhibited the strongest synergistic effect in scavenging DPPH radicals and reducing ferric ions (P<0.05). Moreover, E1 presented strong cytoprotection against H2O2-induced oxidative damage in MRC-5 cells by suppressing the decrease of the superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) activities. A strong correlation between the increment of total phenolic/flavonoid and synergistic antioxidant activity, especially between the increment of total flavonoid and the increase in ferric reducing power was observed. Finally, seven antioxidant substances were identified in E1 as oxypaeoniflora, catechin, calycosin-7-O-β-D-glucopyranoside, fomononetin-7-O-β-D-glucopyranoside, 9,10-dimethoxy-pterocarpan-3-O-β-D-glucopyranoside, quercetin and 2'-dihydroxy-3',4'-dimethyl-isoflavan-7-O-β-D-glucopyranoside.

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HPLC-DAD chromatograms of E1 before and after reaction with DPPH radicals.
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pone-0096780-g004: HPLC-DAD chromatograms of E1 before and after reaction with DPPH radicals.

Mentions: The on-line DPPH-HPLC method has been widely used for screening of the potential radical-scavenging components from various food extracts [30]. After reaction with DPPH, the peak areas (PAs) of the radical scavenging compounds would obviously disappear or decrease in UV chromatograph, while for those without antioxidant activity, there was almost no change in their PAs. Herein, this approach was employed to screen the potential antioxidant compounds from E1. HPLC profiles of E1 with or without DPPH treatment are illustrated in Fig. 4. It was found that eight peaks (1, 3, 4, 6, 7, 9, 10 and 20) were decreased in the PA after spiking with the DPPH free radicals, suggesting these components could be regarded as the potential antioxidant candidates of E1. Among the identified peaks, peaks 1, 3 and 4 were present in PL, while the others (6, 7, 9, 10 and 20) were from AME. Further characterization of these potential antioxidant components was carried out by HPLC-MS/MS, owing to it can provide affluent multistage fragment information for compounds with collision-induced dissociation [31]. As shown in Table 3, seven antioxidant components were unambiguously identified as Oxypaeoniflora (1), Catechin (3), Calycosin-7-O-β-D-glucopyranoside (6), Fomononetin-7-O-β-D-glucopyranoside (7), 9,10-dimethoxy-pterocarpan-3-O-β-D-glucopyranoside (9), 2′-dihydroxy-3′,4′-dimethyl–isoflavan-7-O-β-D-glucopyranoside (10), Quercetin (20), by comparing their UV maximum absorption wavelength, MS and MS/MS data with those of the standard references, as well as the previous reports [32], [33]. It is not surprising that most of these components belonged to the flavonoids family, which has been confirmed to be excellent antioxidants.


In vitro synergistic antioxidant activity and identification of antioxidant components from Astragalus membranaceus and Paeonia lactiflora.

Xu X, Li F, Zhang X, Li P, Zhang X, Wu Z, Li D - PLoS ONE (2014)

HPLC-DAD chromatograms of E1 before and after reaction with DPPH radicals.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4016014&req=5

pone-0096780-g004: HPLC-DAD chromatograms of E1 before and after reaction with DPPH radicals.
Mentions: The on-line DPPH-HPLC method has been widely used for screening of the potential radical-scavenging components from various food extracts [30]. After reaction with DPPH, the peak areas (PAs) of the radical scavenging compounds would obviously disappear or decrease in UV chromatograph, while for those without antioxidant activity, there was almost no change in their PAs. Herein, this approach was employed to screen the potential antioxidant compounds from E1. HPLC profiles of E1 with or without DPPH treatment are illustrated in Fig. 4. It was found that eight peaks (1, 3, 4, 6, 7, 9, 10 and 20) were decreased in the PA after spiking with the DPPH free radicals, suggesting these components could be regarded as the potential antioxidant candidates of E1. Among the identified peaks, peaks 1, 3 and 4 were present in PL, while the others (6, 7, 9, 10 and 20) were from AME. Further characterization of these potential antioxidant components was carried out by HPLC-MS/MS, owing to it can provide affluent multistage fragment information for compounds with collision-induced dissociation [31]. As shown in Table 3, seven antioxidant components were unambiguously identified as Oxypaeoniflora (1), Catechin (3), Calycosin-7-O-β-D-glucopyranoside (6), Fomononetin-7-O-β-D-glucopyranoside (7), 9,10-dimethoxy-pterocarpan-3-O-β-D-glucopyranoside (9), 2′-dihydroxy-3′,4′-dimethyl–isoflavan-7-O-β-D-glucopyranoside (10), Quercetin (20), by comparing their UV maximum absorption wavelength, MS and MS/MS data with those of the standard references, as well as the previous reports [32], [33]. It is not surprising that most of these components belonged to the flavonoids family, which has been confirmed to be excellent antioxidants.

Bottom Line: Many traditionally used herbs demonstrate significantly better pharmacological effects when used in combination than when used alone.Among them, E1 exhibited the strongest synergistic effect in scavenging DPPH radicals and reducing ferric ions (P<0.05).A strong correlation between the increment of total phenolic/flavonoid and synergistic antioxidant activity, especially between the increment of total flavonoid and the increase in ferric reducing power was observed.

View Article: PubMed Central - PubMed

Affiliation: Department of Food Science, Shandong Agricultural University, Taian, China.

ABSTRACT
Many traditionally used herbs demonstrate significantly better pharmacological effects when used in combination than when used alone. However, the mechanism underlying this synergism is still poorly understood. This study aimed to investigate the synergistic antioxidant activity of Astragalus membranaceus (AME) and Paeonia Lactiflora (PL), and identify the potential antioxidant components by 1,1-diphenyl-2-picrylhydrazine (DPPH) radical spiking test followed by a high performance liquid chromatography separation combined with diode array detection and tandem mass spectrometry analysis (DPPH-HPLC-DAD-MS/MS). Eight AME-PL combined extracts (E1-E8) were prepared based on bioactivity-guided fractionation. Among them, E1 exhibited the strongest synergistic effect in scavenging DPPH radicals and reducing ferric ions (P<0.05). Moreover, E1 presented strong cytoprotection against H2O2-induced oxidative damage in MRC-5 cells by suppressing the decrease of the superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) activities. A strong correlation between the increment of total phenolic/flavonoid and synergistic antioxidant activity, especially between the increment of total flavonoid and the increase in ferric reducing power was observed. Finally, seven antioxidant substances were identified in E1 as oxypaeoniflora, catechin, calycosin-7-O-β-D-glucopyranoside, fomononetin-7-O-β-D-glucopyranoside, 9,10-dimethoxy-pterocarpan-3-O-β-D-glucopyranoside, quercetin and 2'-dihydroxy-3',4'-dimethyl-isoflavan-7-O-β-D-glucopyranoside.

Show MeSH
Related in: MedlinePlus