Dehydroascorbic acid dha

Manufactured by Merck Group

Sourced in United States, Sao Tome and Principe

Dehydroascorbic acid (DHA) is a chemical compound that is the oxidized form of ascorbic acid (vitamin C). DHA is a key intermediate in the biosynthesis and catabolism of ascorbic acid. It serves as a sensitive indicator of oxidative stress and redox status in biological systems.

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5 protocols using dehydroascorbic acid dha

Carotenoid and Phenolic Analysis

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Acetone (HPLC grade) from Sigma–Aldrich (Poland, Warsaw); diethyl ether (HPLC grade) from Merck; hexane (HPLC grade) from Sigma–Aldrich (Poland, Warsaw); methanol (HPLC grade) from Sigma–Aldrich; carotenoid standards (HPLC grade 99.5–99.9% pure) from Sigma–Aldrich; β-carotene, lutein, zeaxanthin and magnesium carbonate (pure) from ChemPur; sodium carbonate (analytical grade) from ChemPur; sodium chloride (analytical grade) from Sigma–Aldrich; and sodium peroxide (analytical grade) from ChemPur were used, with phenolic standards (purity 99.0–99.9%) from Sigma–Aldrich, and L-ascorbic acid (L-Asc) and dehydroascorbic acid (DHA) standards with 99% purity (Sigma–Aldrich, Poznan, Poland).

Ponder A., Jariené E, & Hallmann E. (2022). The Effect of Storage Conditions on the Content of Molecules in Malus domestica ‘Chopin’ cv. and Their In Vitro Antioxidant Activity. Molecules, 27(20), 6979.

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Foliar Application of Chemical Treatments

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The following chemicals with respective concentrations were used as: reduced ascorbic acid (AA; Sigma-Aldrich, Missouri, United States) at 20 mM (Singh et al., 2020b (link)), ascorbate oxidase (AO; Sigma-Aldrich, Missouri, United States) at 20 U/ml (Singh et al., 2020b (link)), dehydroascorbic acid (DHA, Sigma-Aldrich, Missouri, United States) at 20 mM (Singh et al., 2020b (link)), methyl jasmonate (MeJA, Sigma-Aldrich) at 100 μM (Nahar et al., 2011 (link)), and AOPP, an inhibitor of PAL activity, at 100 μM (Ji et al., 2015 (link); Khanam et al., 2018 (link)). These concentrations have been optimized in the previous publications with chemical concentrations tested for bio-efficacy and lack of phytotoxicity. AOPP was dissolved in 1 ml of EtOH before diluting further in water. All other chemicals were dissolved in water. To allow efficient uptake, all solutions were supplemented with 0.02% (v/v) of Tween20 (Nahar et al., 2011 (link)) and were then administered via foliar spraying using Fantasea spray bottles (Jojoli, Netherlands). This allows spraying a fine mist without clogging. In each experiment, 14-day-old plants were sprayed with 6.25 ml of solution until runoff. Foliar spraying was done 24 h prior to nematode or mock inoculation. Plants in the control group were mock sprayed with distilled water containing 0.02% (v/v) of Tween20.

Singh R.R., Pajar J.A., Audenaert K, & Kyndt T. (2021). Induced Resistance by Ascorbate Oxidation Involves Potentiating of the Phenylpropanoid Pathway and Improved Rice Tolerance to Parasitic Nematodes. Frontiers in Plant Science, 12, 713870.

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Piperonylic Acid and Dehydroascorbic Acid Treatment on Rice Plants

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Piperonylic acid (PA; Sigma-Aldrich) was prepared as a 100 mM stock solution in DMSO, and diluted with distilled water containing 0.1 vol% Tween 20 surfactant to a final concentration of 300 µM. Dehydroascorbic acid (DHA; Sigma-Aldrich) was prepared by dissolving frozen DHA in distilled water with 0.1 vol% Tween 20 to a concentration of 20 mM. Mock-treated plants were sprayed with distilled water containing the same concentration of Tween 20 and DMSO.
Rice plants were sprayed with an atomizer until run-off (approximately 2 ml per seedling).

Desmedt W., Kudjordjie E.N., Chavan S.N., Zhang J., Li R., Yang B., Nicolaisen M., Mori M., Peters R.J., Vanholme B., Vestergård M, & Kyndt T. (2022). Rice diterpenoid phytoalexins are involved in defence against parasitic nematodes and shape rhizosphere nematode communities. The New phytologist, 235(3).

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Characterization of Recombinant GSTO2 Enzyme

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rGSTO2 activities were determined by spectrophotometry using a panel of substrates [21 ]. The GST activity of 1-chloro-2, 4-dinitrobenzene (CDNB, Sigma-Aldrich) on rGSTO2 protein was measured at 340 nm absorbance. The reaction system was composed of 0.6 mM CDNB, 6 mM glutathione (GSH, Sigma-Aldrich) and 10 μg rGSTO2 protein, and the reaction was carried out in 100 mM phosphate buffer (pH 7.4). Furthermore, the activity of dehydroascorbate reductase (DHAR) was assayed in potassium phosphate buffer (50 mM, pH 7.4) containing 1 mM GSH, 0.25 mM dehydroascorbic acid (DHA, Sigma-Aldrich) and 10 μg rGSTO2 protein [39 (link)]. Thioltransferase activity was analyzed in potassium phosphate buffer (50 mM, pH 7.4) containing 0.2 mM NADPH, 0.5 mM GSH, 2 mM hydroxyethyl disulfide (HED) and 10 μg rGSTO2 protein [3 (link)]. The maximum reaction rate (V_max) and Michaelis constant (K_m) at the same temperature and pH were calculated according to the Michaelis–Menten equation [36 (link)].

Wang X., Zhao C., Zhang G., Zhang K., Li Z., Shang Y., Ning C., Ji C., Xia X., Cai X., Qiao J, & Meng Q. (2022). Molecular characterization of a novel GSTO2 of Fasciola hepatica and its roles in modulating murine macrophages. Parasite, 29, 16.

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Quantification of Ascorbic and Dehydroascorbic Acids

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Reagents Crystalline ascorbic acid (AA) (99%), dehydroascorbic acid (DHA) 99%, DL-dithiothreitol 99%, and ethyl acetate 99.8% were purchased from Sigma-Aldrich (St. Louis, MO, USE). Malic acid, metaphosphoric acid (40-50%), tris(hydroxymethyl)aminomethane, and di-hydrated monobasic sodium phosphate were obtained from VWR Prolabo chemicals (Luther Worth, England). Sodium hydroxide (97%), di-hydrated dibasic sodium phosphate and hydrochloric acid 37% came from Carlo Erba (Val de Reuil, France). Water used to prepare the solutions and dilutions was purified using an ELIX system (Millipore, Bedford, MA).

Gómez Ruiz B., Roux S., Courtois F, & Bonazzi C. (2018). Kinetic modelling of ascorbic and dehydroascorbic acids concentrations in a model solution at different temperatures and oxygen contents. Food research international (Ottawa, Ont.), 106.

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