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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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Ferric reducing antioxidant power of eight combinations of CF-AME and chromatographic fractions from EA-PL (a), and correlation between the rate of increase in total phenolic/flavonoid (RIFC/RIPC), CIs and RIFrapV values (b).(a-A) FRAP value; (a-B) RIFrapV is defined as the rate of increase in FRAP value; (b-A) Correlation between the RIFC/RIPC and CI values; (b-B) Correlation between RIFC/RIPC and RIFrapV values. Solid symbols represent the correlation between RIFC and CI or RIFrapV values, while open symbols represent the correlation between RIPC and CI or RIFrapV values.
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pone-0096780-g003: Ferric reducing antioxidant power of eight combinations of CF-AME and chromatographic fractions from EA-PL (a), and correlation between the rate of increase in total phenolic/flavonoid (RIFC/RIPC), CIs and RIFrapV values (b).(a-A) FRAP value; (a-B) RIFrapV is defined as the rate of increase in FRAP value; (b-A) Correlation between the RIFC/RIPC and CI values; (b-B) Correlation between RIFC/RIPC and RIFrapV values. Solid symbols represent the correlation between RIFC and CI or RIFrapV values, while open symbols represent the correlation between RIPC and CI or RIFrapV values.

Mentions: The antioxidant activity of the eight combined extracts (E1–E8) was estimated by the FRAP assay, in which the antioxidants present in the sample reduce the Fe(III)/tripyridyltriazine (TPTZ) complex to the blue ferrous form, with an increase in absorbance at 593 nm. In order to obtain FRAP values, a calibration curve generated by analyzing standard FeSO4 solutions was derived from the absorbance versus concentration plot (R2 = 0.9979). RIFrapV was also calculated as the rate of increase in FRAP value in the combined extracts as compared to theoretical sum of those of the respective herbs extract. As shown in Fig.3a-A, the highest FRAP value was observed for E1 (3.392±0.015 mmol Fe2+/g DW), suggesting that it had the strongest ferric reducing ability. The FRAP values for other samples decreased in the order: E4>E5>E6>E2>E3, E7, E8. As to RIFrapV, E1 exhibited a highest RIFrapV value of 229.68% (Fig.3a-B), showing that there was a synergistic enhancement in ferric reducing ability in E1. Additionally, the antioxidant activity of eight combined extracts, based on FRAP assay, was different from that based on the DPPH scavenging test. It might be explained by considering that these determinations were performed under different experimental conditions which were based on different reaction mechanisms. DPPH assay is believed to be one of the methods utilizing both hydrogen atom transfer and single electron transfer mechanism. The FRAP assay takes advantage of electron-transfer reaction and takes place at a faster rate than that for the DPPH test, whose degree of discoloration is attributed to the hydrogen donating ability of the test compounds [25].


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)

Ferric reducing antioxidant power of eight combinations of CF-AME and chromatographic fractions from EA-PL (a), and correlation between the rate of increase in total phenolic/flavonoid (RIFC/RIPC), CIs and RIFrapV values (b).(a-A) FRAP value; (a-B) RIFrapV is defined as the rate of increase in FRAP value; (b-A) Correlation between the RIFC/RIPC and CI values; (b-B) Correlation between RIFC/RIPC and RIFrapV values. Solid symbols represent the correlation between RIFC and CI or RIFrapV values, while open symbols represent the correlation between RIPC and CI or RIFrapV values.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0096780-g003: Ferric reducing antioxidant power of eight combinations of CF-AME and chromatographic fractions from EA-PL (a), and correlation between the rate of increase in total phenolic/flavonoid (RIFC/RIPC), CIs and RIFrapV values (b).(a-A) FRAP value; (a-B) RIFrapV is defined as the rate of increase in FRAP value; (b-A) Correlation between the RIFC/RIPC and CI values; (b-B) Correlation between RIFC/RIPC and RIFrapV values. Solid symbols represent the correlation between RIFC and CI or RIFrapV values, while open symbols represent the correlation between RIPC and CI or RIFrapV values.
Mentions: The antioxidant activity of the eight combined extracts (E1–E8) was estimated by the FRAP assay, in which the antioxidants present in the sample reduce the Fe(III)/tripyridyltriazine (TPTZ) complex to the blue ferrous form, with an increase in absorbance at 593 nm. In order to obtain FRAP values, a calibration curve generated by analyzing standard FeSO4 solutions was derived from the absorbance versus concentration plot (R2 = 0.9979). RIFrapV was also calculated as the rate of increase in FRAP value in the combined extracts as compared to theoretical sum of those of the respective herbs extract. As shown in Fig.3a-A, the highest FRAP value was observed for E1 (3.392±0.015 mmol Fe2+/g DW), suggesting that it had the strongest ferric reducing ability. The FRAP values for other samples decreased in the order: E4>E5>E6>E2>E3, E7, E8. As to RIFrapV, E1 exhibited a highest RIFrapV value of 229.68% (Fig.3a-B), showing that there was a synergistic enhancement in ferric reducing ability in E1. Additionally, the antioxidant activity of eight combined extracts, based on FRAP assay, was different from that based on the DPPH scavenging test. It might be explained by considering that these determinations were performed under different experimental conditions which were based on different reaction mechanisms. DPPH assay is believed to be one of the methods utilizing both hydrogen atom transfer and single electron transfer mechanism. The FRAP assay takes advantage of electron-transfer reaction and takes place at a faster rate than that for the DPPH test, whose degree of discoloration is attributed to the hydrogen donating ability of the test compounds [25].

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