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PubMed1986

Ascorbate in plasma as measured by liquid chromatography and by dichlorophenolindophenol colorimetry.

Ascorbic acid was measured in 125 plasma samples by an automated colorimetric method involving dichlorophenolindophenol and by a "high-performance" liquid-chromatographic procedure with electrochemical detection. The two methods gave comparable results for samples with ascorbate concentrations of 1 to 20 mg/L (r = 0.97). We also measured the amount of total ascorbate (ascorbic acid + dehydroascorbic acid) in the same samples by a liquid-chromatographic procedure with precolumn derivitization of ascorbic acid. We confirmed that plasma contains little dehydroascorbic acid.

2,6-Dichloroindophenol↗
PubMed1992

Sex variation in ascorbic acid catabolism.

Male and female albino rats of same age and body weight were pair fed with laboratory stock diet and ascorbic acid, dehydroascorbic acid and diketogulonic acid were determined in the liver and urine, while in blood only ascorbic acid was estimated. Male rats had concentration higher of ascorbic acid in liver and urine as compared with females, while there were no significant variations in the contents of dehydroascorbic acid and diketogulonic acid. Hepatic and renal 2, 3-diketoaldonate decarboxylase, and hepatic dehydroascorbatase were also found to be significantly higher in male rats. Similar sex variations were also observed in ascorbic acid catabolism in guinea pigs without any differences in urinary ascorbic acid contents.

Animals↗
PubMed1987

In vitro oxidation of ascorbic acid and its prevention by GSH.

The interaction of glutathione (GSH) with ascorbic acid and dehydroascorbic acid was examined in in-vitro experiments in order to examine the role of GSH in protecting against the autoxidation of ascorbic acid and in regenerating ascorbic acid by reaction with dehydroascorbic acid. If a buffered solution (pH 7.4) containing 1.0 mM ascorbic acid was incubated at 37 degrees C, there was a rapid loss of ascorbic acid in the presence of oxygen. When GSH was added to this solution, ascorbic acid did not disappear. Maximum protection against ascorbic acid autoxidation was achieved with as little as 0.1 mM GSH. Cupric ions (0.01 mM) greatly accelerated the rate of autoxidation of ascorbic acid, an effect that was inhibited by 0.1 mM GSH. Other experiments showed that GSH complexes with cupric ions, resulting in in a lowering of the amount of GSH in solution as measured in GSH standard curves. These results suggest that the inhibition of ascorbic acid autoxidation by GSH involves complexation with cupric ions that catalyze the reaction. When ascorbic acid was allowed to autoxidize at 37 degrees C the subsequent addition of GSH (up to 10 mM) did not lead to the regeneration of ascorbic acid. This failure to detect a direct reaction between GSH and the dehydroascorbic acid formed by oxidation of ascorbic acid under this condition was presumably due to the rapid hydrolysis of dehydroascorbic acid. When conditions were chosen, i.e., low temperature, that promote stability of dehydroascorbic acid, the direct reaction between GSH and dehydroascorbic acid to form ascorbic acid was readily detected. The marked instability of dehydroascorbic acid at 37 degrees C raises questions regarding the efficiency of the redox couple between GSH and dehydroascorbic acid in maintaining the concentration of ascorbic acid in mammalian cells exposed to an oxidative challenge.

Ascorbic Acid↗
PubMed1986

Vitamin stability in a TPN mixture stored in an EVA plastic bag.

In order to examine the stability of vitamins in a TPN admixture stored in 3-litre plastic (EVA) bags, two different stability studies were performed. In the first experiment the TPN admixture was stored in darkness at 2-8 degrees C for 96 h and the stability of vitamins determined. The vitamins examined were retinyl palmitate, alpha-tocopherol, thiamine mononitrate, sodium ascorbate (analysed as reduced ascorbic acid and dehydroascorbic acid), sodium riboflavin-5'-phosphate, pyridoxine hydrochloride, nicotinamide, folic acid, biotin, sodium pantothenate and cyanocobalamin. In the second test the stability of vitamins was determined during simulated infusion from the bag containing the admixture. The vitamins examined were retinyl palmitate, alpha-tocopherol, sodium riboflavin-5'-phosphate and sodium ascorbate (analysed as reduced ascorbic acid and dehydroascorbic acid). The vitamin stability was found to be acceptable for all vitamins except ascorbic acid and folic acid. Total ascorbic acid is the sum of reduced ascorbic acid and dehydroascorbic acid (DHA). It is important to estimate the total ascorbic acid concentration because DHA is also biological active. About 50% of the nominal total ascorbic acid remained after 96 h of storage at 2-8 degrees C in darkness, or after 24 h of simulated infusion initiated immediately after mixing. With folic acid there appears to be assay interference which requires further investigation.

Drug Packaging↗
PubMed1989

Selected indices of micronutrient status in adult patients with sickle cell anemia (SCA).

In 24 adults with hemoglobin SS followed at the Duke University Comprehensive Sickle Cell Center, we have studied the following nutritional parameters: reduced ascorbic acid; dehydroascorbic acid; alpha and beta carotenes; cryptoxanthin; and alpha and gamma tocopherols in whole blood, washed red blood cells, plasma, or serum. In the same population we also examined reduced glutathione (GSH) and oxidized glutathione (GSSG). Fifteen of these 24 patients also were interviewed for usual dietary intakes using a 28-day dietary history. Data obtained from patients with hemoglobin SS, sickle cell anemia (SCA) were compared to those found for seven healthy normal black adults of similar age. Plasma alpha tocopherol levels were significantly lower in SCA individuals than those of the controls (P less than 0.004). Alpha and gamma tocopherol levels in sickle RBCs were significantly higher than those from RBC suspensions of control subjects (P less than 0.007, and P less than 0.001, respectively). All serum values for carotenoids examined, specifically, beta carotene, alpha carotene, and cryptoxanthin were also markedly depressed when compared to those of healthy controls (P less than 0.001, P less than 0.002, and P less than 0.001, respectively). No other statistically significant differences were found between the two groups for any of the remaining variables, including dietary estimates. Dietary analyses suggest that dietary intakes of SCA individuals exceeded the recommended daily allowances (RDA) of all macro- and micronutrients measured, and intakes of most nutrients exceeded those of black controls interviewed. These results suggest that in individuals with SCA, several micronutrients vital to maintaining reducing capacity are present in diminished quantities in plasma/serum. These anomalies exist in SCA patients even though their intake of these micronutrients are similar to those of healthy black men and women.

Adult↗
PubMed1987

[Quality changes in the storage of vegetable peas (Pisum sativum L.). 2. Nutritional physiologic quality].

In tests extending over several years the typical changes in the nutritional quality as occur during storage at defined temperatures (0 ... 18 degrees C) were determined in dependence on the duration of storage (chi in days). Due to the nutrient transfer from the pods to the seeds, a temporary major rise of the dry matter content (DM) is recorded in the latter together with a strongly inhibited reduction of the mono- and disaccharide content. Green peas stored without pods show a linear decrease in the DM content, degressively increasing DM losses and a more distinct reduction of saccharides. When stored with the pods, the typical decline of the vitamin C (ascorbic acid + dehydroascorbic acid) [changes in the content according to the equation y = alpha 0 + beta 1e-c theta chi and losses according to y = beta 1(1 - e-c theta chi)] goes back, mainly owing to an essentially lower coefficient for c theta. In addition, there are smaller losses in the total carotene content of such crops in the first stage of storage (losses of y = beta 1 chi 2). When storing peas without pods, we observe a a faster accumulation of the titratable total acid (up to 160%). The greatest influence on range and degree of the changes in the chemical constituents is exerted by the storage temperature. The form of the harvested crop, varietal characteristics and annual growing conditions have less bearing. Relationships and physiological causes are discussed.

Fabaceae↗
PubMed1989

Manganese catalysis of dopamine oxidation.

Manganese catalysis of the oxidation of dopamine by air was studied as part of an investigation of possible manganese intoxication amongst a group of Aborigines living on manganese-rich soil on Groote Eylandt, in the Northern Territory of Australia. Manganese significantly increased the oxidation rate of dopamine, and the manganese complexes with some purines were especially efficient catalysts. An oxidation mechanism, involving a manganese(II)/(III) redox couple and a semiquinone free radical intermediate, is proposed. Stoichiometric hydrogen peroxide was produced by the oxidation, and the oxidation products of dopamine were highly toxic to the marine diatom Nitzschia closterium. Hydrogen peroxide and the superoxide radical did not oxidize dopamine at physiological pH. Some electrophilic compounds, including ascorbic acid, dehydroascorbic acid and thiamine, effectively inhibited dopamine oxidation. The Groote Eylandt Aborigines are likely to be deficient in ascorbic acid (vitamin C) and thiamine (vitamin B1), and these deficiencies, as well as their lifestyle, may predispose them to manganese intoxication.

Catalysis↗
PubMed1987

Determination of ascorbic acid in human urine by high-performance liquid chromatography coupled with fluorimetry after post-column derivatization with benzamidine.

High-performance liquid chromatography on two Asahipak GS-320 hydrophilic gel columns (50 X 0.76 cm I.D.), connected in series, with 0.015 M tartrate buffer (pH 3.0), containing 2 mM ethylenediaminetetraacetate and 0.05% beta-thiodiglycol as eluent allowed the separation of glucose, diketogulonic acid + diketogluconic acid, dehydroisoascorbic acid, dehydroascorbic acid, ascorbic acid, and isoascorbic acid within 55 min. Ascorbic acid in a urine sample was stabilized by the addition of an equal volume of 5% metaphosphoric acid solution, containing 0.5% of beta-thiodiglycol. Filtration of the mixture through a column of Dowex 50W-X8 (H+) facilitated the determination of ascorbic acid and isoascorbic acid in human urine. Samples could be analyzed every 20 min.

Ascorbic Acid↗