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      Isolation of Strong Antioxidants from Paeonia Officinalis Roots and Leaves and Evaluation of Their Bioactivities

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          Abstract

          Paeonia officinalis extracts from leaves and roots were tested for their antioxidant potential using in vitro chemical (Folin-Ciocalteu, 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), 2,2′-azino- bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), oxygen radical absorbance capacity (ORAC), hydroxyl radical antioxidant capacity (HORAC), hydroxyl radical scavenging capacity HOSC)) and cellular antioxidant activity (CAA) assays. Leaf extracts were stronger antioxidants than root extracts, while methanol was a more effective solvent than water in chemical assays. However, the selected water extract of leaves was a stronger antioxidant in CAA than the methanol extract (0.106 vs. 0.046 µmol quercetin equivalents/mg). Twenty compounds were identified by ultra performance liquid chromatography-quadrupole-time-of-flight (UPLC-Q-TOF) mass spectrometer, while on-line screening of their antioxidant capacity by high performance liquid chromatography (HPLC) with a DPPH -scavenging detector revealed that gallic acid derivatives are the major peony antioxidants. Root water and leaf methanol extracts inhibited α-amylase in a dose dependent manner. The IC 50 value for the strongest inhibitor, the methanol extract of leaves, was 1.67 mg/mL. In addition, the cytotoxicity assessment of extracts using human Caco-2 cells demonstrated that none of them possessed cytotoxic effects.

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          Most cited references43

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          The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics.

          The human intestinal Caco-2 cell line has been extensively used over the last twenty years as a model of the intestinal barrier. The parental cell line, originally obtained from a human colon adenocarcinoma, undergoes in culture a process of spontaneous differentiation that leads to the formation of a monolayer of cells, expressing several morphological and functional characteristics of the mature enterocyte. Culture-related conditions were shown to influence the expression of these characteristics, in part due to the intrinsic heterogeneity of the parental cell line, leading to selection of sub-populations of cells becoming prominent in the culture. In addition, several clonal cell lines have been isolated from the parental line, exhibiting in general a more homogeneous expression of differentiation traits, while not always expressing all characteristics of the parental line. Culture-related conditions, as well as the different Caco-2 cell lines utilized in different laboratories, often make it extremely difficult to compare results in the literature. This review is aimed at summarizing recent, or previously unreviewed, data from the literature on the effects of culture-related factors and the influence of line sub-types (parental vs. different clonal lines) on the expression of differentiation traits important for the use of Caco-2 cells as a model of the absorptive and defensive properties of the intestinal mucosa. Since the use of Caco-2 cells has grown exponentially in recent years, it is particularly important to highlight these methodological aspects in order to promote the standardization and optimisation of this intestinal model.
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            Cellular antioxidant activity (CAA) assay for assessing antioxidants, foods, and dietary supplements.

            A cellular antioxidant activity (CAA) assay for quantifying the antioxidant activity of phytochemicals, food extracts, and dietary supplements has been developed. Dichlorofluorescin is a probe that is trapped within cells and is easily oxidized to fluorescent dichlorofluorescein (DCF). The method measures the ability of compounds to prevent the formation of DCF by 2,2'-azobis(2-amidinopropane) dihydrochloride (ABAP)-generated peroxyl radicals in human hepatocarcinoma HepG2 cells. The decrease in cellular fluorescence when compared to the control cells indicates the antioxidant capacity of the compounds. The antioxidant activities of selected phytochemicals and fruit extracts were evaluated using the CAA assay, and the results were expressed in micromoles of quercetin equivalents per 100 micromol of phytochemical or micromoles of quercetin equivalents per 100 g of fresh fruit. Quercetin had the highest CAA value, followed by kaempferol, epigallocatechin gallate (EGCG), myricetin, and luteolin among the pure compounds tested. Among the selected fruits tested, blueberry had the highest CAA value, followed by cranberry > apple = red grape > green grape. The CAA assay is a more biologically relevant method than the popular chemistry antioxidant activity assays because it accounts for some aspects of uptake, metabolism, and location of antioxidant compounds within cells.
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              Analysis of antioxidant activities of common vegetables employing oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays: a comparative study.

              A total of 927 freeze-dried vegetable samples, including 111 white cabbages, 59 carrots, 51 snap beans, 57 cauliflower, 33 white onions, 48 purple onions, 130 broccoli, 169 tomatoes, 25 beets, 88 peas, 88 spinach, 18 red peppers, and 50 green peppers, were analyzed using the oxygen radical absorption capacity (ORAC) and ferric reducing antioxidant capacity (FRAP) methods. The data show that the ORAC and FRAP values of vegetable are not only dependent on species, but also highly dependent on geographical origin and harvest time. The two antioxidant assay methods, ORAC and FRAP, also give different antioxidant activity trends. The discrepancy is extensively discussed based on the chemistry principles upon which these methods are built, and it is concluded that the ORAC method is chemically more relevant to chain-breaking antioxidants activity, while the FRAP has some drawbacks such as interference, reaction kinetics, and quantitation methods. On the basis of the ORAC results, green pepper, spinach, purple onion, broccoli, beet, and cauliflower are the leading sources of antioxidant activities against the peroxyl radicals.
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                Author and article information

                Journal
                Antioxidants (Basel)
                Antioxidants (Basel)
                antioxidants
                Antioxidants
                MDPI
                2076-3921
                27 July 2019
                August 2019
                : 8
                : 8
                : 249
                Affiliations
                [1 ]Department of Food Science and Technology, Kaunas University of Technology, Radvilėnų˛ pl. 19, LT-50254 Kaunas, Lithuania
                [2 ]IBET—Instituto de Biologia Experimental e Tecnológica, Food & Health Division Apartado 12, 2780-901 Oeiras, Portugal
                Author notes
                [* ]Correspondence: rimas.venskutonis@ 123456ktu.lt ; Tel.: +370-699-40978 or +370-37-456647
                Author information
                https://orcid.org/0000-0003-4792-7340
                https://orcid.org/0000-0002-4888-9414
                https://orcid.org/0000-0002-6063-8132
                Article
                antioxidants-08-00249
                10.3390/antiox8080249
                6721766
                31357649
                70e7e258-7530-49c0-a9b2-d094ee8b0ca8
                © 2019 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 27 June 2019
                : 25 July 2019
                Categories
                Article

                p. officinalis,phytochemicals,antioxidant capacity,cytotoxicity,α-amylase inhibition

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