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[Reevaluation of optimum extraction condition for urinary organic acids using a stable isotope dilution technique].

We reevaluated the optimum conditions for analysis of 13 urinary organic acids using solvent extraction and GC/MS by the stable isotope dilution technique. The acids analyzed were uracil, and lactic, oxalic, 3-hydroxybutyric, succinic, fumaric, glutaric, adipic, pyroglutamic, 2-ketoglutaric, orotic, sebacic, and citric acids. Analytical recovery and accuracy for 13 organic acids ranged from 90 to 107% and from 1.6 to 13.7%, respectively. The optimal pH for most organic acids was 1-2, while that for oxalic and citric acids was 0.25, and that for 2-ketoglutaric and orotic acids was 0.5. The presence of urinary albumin decreased the extraction rates of organic acids, especially of orotic and citric acids; in slight albuminuria (0.5 g/l) the extraction recovery (the extraction rate with albumin/that without albumin) of orotic and citric acid was 38% and 67%, respectively, and in more marked albuminuria (5 g/l), 6% and 26%. Membrane pretreatment with Centricon-3 improved these extraction rates under the condition of albuminuria. Dehydration with the desiccant agent decreased urinary acid extraction rates, especially of uracil and orotic and citric acids. The extraction rates of these three organic acids was decreased in albuminuria and by the desiccant agent. Accurate quantitative analysis of urinary organic acids by the stable isotope dilution technique is necessary for routine examination conducted in clinical laboratories.

Acids↗

Monitoring bacterial metabolites in cultured buttermilk by high performance liquid chromatography and headspace gas chromatography.

A commercial buttermilk culture was added to Grade a skim milk. Samples were analyzed hourly for a total of eighteen hours. Headspace gas chromatography measured changes in acetaldehyde, acetone, ethanol, and diacetyl concentrations. High performance liquid chromatography measured changes in orotic, citric, pyruvic, lactic, uric, acetic, and propionic acid concentrations. Consumption of orotic acid and citric acid substrates was noted. Production of diacetyl, ethanol, pyruvic, lactic, acetic, and propionic acids was observed. Acetaldehyde concentration increased initially and then decreased as ethanol was formed. Acetone and uric acid concentrations remained constant during fermentation. In addition, chemical changes occurring during refrigeration were measured after seven days. Acetaldehyde, ethanol, and acetic acid increased in concentration; diacetyl, orotic, citric, pyruvic, lactic, and propionic acids decreased in concentration. Also, a headspace gas chromatographic test was done for the quantitation of acetoin.

Animals↗

Inhibitory action of orotate, 2-thioorotate and isoorotate on nucleotide metabolism and nucleic acid synthesis in hepatoma cells.

1. The specificity of the action of orotate on hepatoma cells was investigated. 2. Orotic acid and its methyl ester had similar inhibitory effects on the incorporation of [3H]thymidine into DNA of hepatoma cells. 3. In contrast to previous studies in vivo, incubation of rat kidney cells with orotate caused an increase in the ratio of UTP/ATP concentrations that was similar to effects on hepatic cells. 4. Inhibitory effects of 2-thioorotate and isoorotate on metabolism were found to be less selective and required higher concentrations than with orotate.

Animals↗