A flavonoid-rich extract of Hypericum perforatum L. (FEHP) was prepared by adsorption on macroporous resin and desorption by ethanol. Total flavonoid content of FEHP was determined by a colorimetric method. The major constituents of FEHP, including rutin, hyperoside, isoquercitrin, avicularin, quercitrin, and quercetin, were determined by HPLC analysis and confirmed by LC-MS. Different antioxidant assays were utilized to evaluate free radical scavenging activity and antioxidant activity of FEHP. FEHP was an effective scavenger in quenching DPPH and superoxide radical with IC50 of 10.63 microg/mL and 54.3 microg/mL, respectively. A linear correlation between concentration of FEHP and reducing power was observed with a coefficient of r2 = 0.9991. Addition of 150 microg of FEHP obviously decreased the peroxidation of linoleic acid during 84 h incubation, but the amount of FEHP over 150 microg did not show statistically significant inhibitory effect of peroxidation of linoliec acid (p > 0.05). FEHP exhibited inhibitory effect of peroxidation of liposome induced both by hydroxyl radical generated with iron-ascorbic acid system and peroxyl radical and showed prominent inhibitory effect of deoxyribose degradation in a concentration-dependent manner in site-specific assay but poor effect in non-site-specific assay, which suggested that chelation of metal ion was the main antioxidant action. According to the results obtained in the present study, the antioxidant mechanism of FEHP might be attributed to its free radical scavenging activity, metal-chelation activity, and reactive oxygen quenching activity.
Phenolic compounds were extracted from Morton lentils using acidified aqueous acetone. The crude Morton extract (CME) was applied onto a macroresin column and desorbed by aqueous methanol to obtain a semi-purified Morton extract (SPME). The SPME was further fractionated over Sephadex LH-20 column into five main fractions (Fr I – Fr V). The phytochemical contents such as total phenolic content (TPC), total flavonoid content (TFC), and condensed tannin content (CTC) of the CME, SPME, and its fractions were examined by colorimetric methods. Antioxidant activity of extracts and fractions were screened by DPPH scavenging activity, trolox equivalent antioxidant capacity (TEAC), ferric reduced antioxidant power (FRAP), and oxygen radical absorbing capacity (ORAC) methods. In addition, the compositions of active fractions were determined by HPLC-DAD and HPLC-MS methods. Results showed that fraction enriched in condensed tannins (Fr V) exhibited significantly higher value of TPC, CTC and higher antioxidant activity as compared to the crude extract, SPME and low-molecular-weight fractions (Fr I – IV). Eighteen compounds existed in those fractions, and seventeen were tentatively identified by UV and MS spectra. HPLC-MS analysis revealed Fr II contained mainly kaempferol glycoside, Fr III and Fr IV mainly contained flavonoid glycosides, and Fr V was composed of condensed tannins. The results suggested that extract of Morton lentils is a promising source of antioxidant phenolics, and may be used as a dietary supplement for health promotion.
Germination is considered to be an effective process for improving the nutritional quality and functionality of cereals. In this study, changes of nutritional ingredients, antinutritional components, chemical composition, and antioxidant activities of buckwheat seeds over 72 h of germination were investigated, and the reasons for these changes are discussed. With the prolonged germination time, the contents of crude protein, reducing sugar, total phenolics, total flavonoids, and condensed tannins increased significantly, while the levels of crude fat, phytic acid, and the activity of trypsin inhibitor decreased. Phenolic compounds, such as rutin, vitexin, isovitexin, orientin, isoorientin, chlorogenic acid, trans-3-hydroxycinnamic acid, and p-hydroxybenzoic acid increased significantly during the germination process, which may be due to the activation of phenylalanine ammonialyase. The improvement of flavonoids led to significant enhancement of the antioxidant activities of germinated buckwheat. Germinated buckwheat had better nutritional value and antioxidant activities than ungerminated buckwheat, and it represented an excellent natural source of flavonoids and phenolic compounds, especially rutin and C-glycosylflavones. Therefore, germinated buckwheat could be used as a promising functional food for health promotion.
Three phlorotannins, including phloroglucinol, diphlorethohydroxycarmalol, and 6,6'-bieckol, were isolated from Ishige okamurae by column chromatography. The structures of the phlorotannins were determined on the basis of spectroscopic analysis, including NMR and mass spectrometry (MS) techniques. Antioxidant effects of phlorotannins were measured by direct free radical scavenging activities using the electron spin resonance spectrometry (ESR) technique and cellular systems in vitro. The results indicated that diphlorethohydroxycarmalol and 6,6'-bieckol showed potential radical scavenging activities against the 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl, alkyl, and superoxide radicals. Moreover, no cytotoxicities of the phlorotannins on human fetal lung fibroblasts cell line (MRC-5), mouse macrophages cell line (RAW264.7), and human leukemic cell line (HL-60) were observed. In addition, diphlorethohydroxycarmalol and 6,6'-bieckol significantly reduced the intracellular reactive oxygen species level assessed by 2',7'-dichlorofluorescein diacetate (DCFH-DA) assay in RAW264.7 cells, and myeloperoxide (MPO) activity in HL-60 cells and radical-mediated oxidation of cell membrane proteins in RAW264.7 cells were dose-dependently inhibited in the presence of diphlorethohydroxycarmalol and 6,6'-bieckol. In conclusion, these results suggested that phlorotannins could be used as novel functional foodstuffs or antioxidants in the cosmetic and drug industries.
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