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At least 19 recordsLinked to original sources

Separation of ascorbic acid, dehydroascorbic acid, diketogulonic acid and glucose by isocratic elution from a column of a hydrophilic gel.

High-performance liquid chromatography on an Asahipak GS-320 hydrophilic gel column with tartrate buffer (0.015 M, pH 3.0) containing 2 mM ethylenediaminetetraacetate and 0.05% beta-thiodiglycol as the eluent allowed the separation of glucose, diketogulonic acid, dehydroascorbic acid and ascorbic acid within 30 min. Fluorimetric monitoring of these compounds in the eluate with benzamidine at alkaline pH and at 90 degrees C in the presence of potassium sulphite allowed the determination of nanogram amounts of ascorbic acid, dehydroascorbic acid and diketogulonic acid. This method was applied to the determination of ascorbic acid in fruit juice.

2,3-Diketogulonic Acid↗

Ascorbic acid, dehydroascorbic acid and glutathione in liver disease.

Controlled studies were conducted to find out the plasma values of ascorbic acid, dehydroascorbic acid (DHA), urinary excretion of ascorbic acid and blood levels of glutathione in patients with viral hepatitis, alcoholic hepatitis, cirrhosis of liver and carcinoma of liver. Leucocyte ascorbic acid and DHA/AA index were also determined in order to assess the ascorbic acid status of these patients. It was observed that the plasma and leucocytes contents of ascorbic acid were significantly subnormal with markedly decreased urinary excretion in these patients. Decreased level of glutathione and significantly higher level of DHA reflect an over all reducing status of the body is markedly deranged in these conditions. Further it was observed that the DHA/AA ratios were significantly altered in these groups of patients.

Ascorbic Acid↗

Intestinal transport and metabolism of oxidized ascorbic acid (dehydroascorbic acid).

The transport and metabolic properties of dehydroascorbic acid have been evaluated in the small intestine of one ascorbic acid-dependent animal species (guinea pig) and one ascorbic acid-synthesizing animal species (rat). Loops of guinea pig jejunum in vitro brought about net transepithelial absorption of L-[14C]dehydroascorbic acid with most of the absorbed substrate appearing in the reduced form (ascorbic acid). Saturation of brush-border transport occurs at a lower concentration than saturation of the enzymatic reduction. Rat intestine did not bring about net transepithelial transport. The intestines of both guinea pigs and rats took up substantial dehydroascorbic acid from the serosal bathing solution, with the majority appearing in the mucosa as ascorbic acid. It is proposed that in guinea pig intestinal mucosa, dehydroascorbic acid reductase has a nutritional role in extracting the vitamin from chyme and reducing it for subsequent use in the body. The intestines of both guinea pig and rat appear to take up dehydroascorbic acid across the basolateral surface and reduce it, thus maintaining a substantial endogenous level of ascorbic acid in the mucosa. The transport and metabolic properties described might play an important role as a protective antioxidant mechanism in intestinal mucosa.

Animals↗

Antioxidant and prooxidant effects of ascorbic acid, dehydroascorbic acid and flavonoids on LDL submitted to different degrees of oxidation.

UNLABELLED: Although a high intake of antioxidants may decrease the risk of developing cardiovascular diseases, under certain circumstances they may promote free radical generation and lipid peroxidation. The objectives of the present study were to determine the antioxidant effects of ascorbic acid (AA), dehydroascorbic acid (DHA) and flavonoids on LDL submitted to different degrees of oxidation. LDL was submitted to oxidation with CuCl2 (2.4 microM). Before or at different times after the propagation of the oxidation process, 28 microM (5 micrograms/ml) of either AA or DHA or 5 micrograms/mL flavonoids extract were added. Alpha-tocopherol, conjugated dienes, thiobarbituric acid reacting substances (TBARS) and LDL electrophoretic mobility were determined as indices of LDL oxidation. The presence of any of the three antioxidants from the onset of the incubation delayed the oxidation process. However, the addition of both DHA and flavonoids to the oxidation process when it was already initiated and alpha-tocopherol consumed, accelerated the oxidation. In contrast, AA delayed the oxidation process even when added after alpha-tocopherol was consumed. Nevertheless, it also accelerated LDL oxidation when added during the propagation phase of the oxidation process. IN CONCLUSION: although AA, DHA and flavonoids delay LDL oxidation when added before the initiation of the process, they accelerate the process if added to minimally oxidized LDL.

Adult↗

Applications of a simultaneous assay of ascorbic acid, dehydroascorbic acid and ascorbic sulphate in biological materials.

A modified spectrophotometric assay for ascorbic acid and its derivatives based on their reaction with 2,4-dinitrophenylhydrazine (DNPH) is described. Using standard ascorbic acid or ascorbic sulphate solutions, together with animal tissue or compound diet extracts, the conditions for ascorbic acid degradation were determined. For the differential measurement of reduced ascorbic acid (AA), dehydroascorbic acid (dAA) and ascorbic sulphate (AS), five series of simultaneous determinations were performed. These included the use of (1) KBrO3 for the hydrolysis of AS, (2) 2,6-dichlorophenolindophenol as an oxidant, (3) DNPH to form a hydrazone derivative with dAA and (4 and 5) two blanks (where ascorbate was degraded) to correct for interfering substances. A variety of vertebrate and invertebrate tissues were examined for their ascorbate content, and the advantages of the modified procedure over currently available assays are discussed. The results suggest that the Artemia cyst is a unique material in which ascorbic sulphate is present in large amounts whereas fish tissues do not contain this form of vitamin C.

Animals↗

Spontaneous conversion of L-dehydroascorbic acid to L-ascorbic acid and L-erythroascorbic acid.

Dehydroascorbic acid, an oxidation product of ascorbic acid (vitamin C), spontaneously decomposed at neutral and higher pH levels to form three products that could be quantitated by HPLC-electrochemical analysis. One of the products was ascorbic acid, suggesting that dehydroascorbic acid was reduced to ascorbic acid without adding an exogenous reductant. The major newly produced compound was almost identical to ascorbic acid by UV spectroscopy, which therefore potentially interfered in the study of ascorbic acid metabolism. The ascorbic acid-like compound was isolated by reversed-phase HPLC and identified as L-erythroascorbic acid by mass spectrometry. Fe(II) and Cu(I) increased, whereas desferrioxamine, a potent iron chelator, inhibited L-erythroascorbic acid production. Phosphate, used as buffer, and cyanide greatly enhanced dehydroascorbic acid conversion to L-erythroascorbic acid. The identification of L-erythroascorbic acid and its quantitation by an electrochemical method provides a useful tool for future study of dehydroascorbic acid metabolism.

Ascorbic Acid↗

Ascorbic acid and dehydroascorbic acid interconversion without net oxidation or reduction.

The interconversion of ascorbic acid and dehydroascorbic acid was examined in aqueous solution using unlabeled dehydroascorbic acid and nonlabile, stable isotope-labeled ascorbic acid by gas chromatographic/mass spectrometric analysis. Although the formation of unlabeled ascorbic acid from unlabeled dehydroascorbic acid or labeled dehydroascorbic acid from labeled ascorbic acid did not occur to any significant extent when either solutions of unlabeled dehydroascorbic acid or labeled ascorbic acid were incubated alone, significant amounts of both labeled dehydroascorbic acid and unlabeled ascorbic acid formed when unlabeled dehydroascorbic acid was incubated with labeled ascorbic acid at acid pH. At alkaline pH, interconversion did not occur to any appreciable extent. Likewise, interconversion did not appear to occur in plasma at physiologic concentrations of ascorbic acid, but did occur with pharmacologic concentrations. These data show that ascorbic acid and dehydroascorbic acid interconvert in acidic solution, suggesting the reducing hydrogen atoms are delocalized when ascorbic acid is paired with dehydroascorbic acid under these circumstances. Alkaline pH and plasma inhibit the interconversion.

Ascorbic Acid↗

Physiological role of dehydroascorbic acid.

Dehydroascorbic acid is present in insignificant amounts in plant and animal tissue but appears in considerable amounts under various physiological and pathological conditions. It is found increased: in blood of patients suffering from infectious diseases; in blood and tissues of thyrotoxic patients; in blood after injection of thyroxin, corticotropin and cortisone. In all the above conditions there is concomitant decrease in L-ascorbic acid and glutathione values of blood and tissues. Dehydroascorbic acid, however, disappears after continued administration of a high dose of ascorbic acid. The accumulation of dehydroascorbic acid seems to be an indication of ascorbic acid deficiency. The extreme sensitivity of the ascorbate system to physiological changes is suggestive of a major biochemical role for this redox system. Accumulated evidences indicate that dehydroascorbic acid possible control cell division.

Animals↗

Determination of ascorbic acid and dehydroascorbic acid in plasma by high-performance liquid chromatography with coulometric detection--are they reliable biomarkers of oxidative stress?

The concentrations of the hydrophilic antioxidants ascorbic acid and dehydroascorbic acid in plasma for some time have been considered possible biomarkers of oxidative stress. However, several problems are associated with the accurate measurement of these two compounds. We have developed and validated a selective and reproducible high-performance liquid chromatographic method for the quantification of ascorbic acid and dehydroascorbic acid in plasma. The method meets the requirements of a reliable routine analysis. The plasma samples are stabilized with 5 mM metaphosphoric acid, centrifugated at 4 degrees C before HPLC analysis. For ascorbic acid analysis, the sample pH is adjusted to 2.6, whereas for total ascorbic acid measurement dehydroascorbic acid is reduced to ascorbic acid using dithiothreitol for 5 min at pH 6.2 after which the sample pH is adjusted to 2.6. The samples are analyzed on a reversed-phase system using coulometric detection. Dehydroascorbic acid concentrations ae calculated by subtraction. Within- and between-day coefficients of variation for the complete assay were in the range of 4-8 and 3-6% for ascorbic acid and total ascorbic acid, respectively. The stability of ascorbic acid was monitored under various conditions including storage and the implications as well as the reliability of ascorbic acid as a biomarker are discussed.

Ascorbic Acid↗