Sample preparation for chromatography of amino acids.
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We measured acetate concentrations in whole blood, serum, and plasma by a modification of a previously described method involving vacuum distillation and gas chromatography. The mean acetate concentration of fresh venous plasma from 27 normal subjects was 51 +/- 5 mumol/L (95% confidence limits ranged from 0 to 103 mumol/L). The acetate concentrations of serum and plasma incubated for 2 h at either 4 degrees C or 27 degrees C were the same. The acetate concentration of whole blood incubated at 27 degrees C was significantly greater than that of blood incubated at 4 degrees C. This change may have resulted from the production of acetate by erythrocytes or from the hydrolysis of acetate esters. Storage of plasma at -20 degrees C for 24 h significantly increased acetate concentrations from 26 +/- 6 mumol/L to 63 +/- 4 mumol/L. After the subjects consumed a standard breakfast, venous plasma acetate concentrations increased from 58 to 97 mumol/L at 30 min. Acetate concentrations in arterial plasma exceeded those in venous plasma. Plasma acetate concentrations were not significantly altered in patients with malignancy or diabetes mellitus, but severe liver disease and severe acidosis were both associated with increased acetate concentrations. These preliminary observations suggest that plasma acetate concentrations may be altered in several disease states.
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We describe and evaluate a method for determining nickel in plasma and urine by atomic absorption spectrometry. Proteins are precipitated with trichloroacetic acid and sulfuric acid; ammonium pyrrolidinedithlocarbamate is used as the chelating agent for nickel, and methyl isobutyl ketone as extraction solvent. The results were compared with results obtained by the acid-digestion technique for removing proteins and other organic substances. Analyses for both plasma and urine were better by the present procedure. The mean and standard deviation for nickel in plasma from 15 healthy individuals was 2.13 +/- 0.58 microgram/liter by this method. For nickel in urine from 15 healthy men the mean and standard deviation was 4.45 +/- 1.9 microgram/liter. The coefficient of variation for plasma was 11.9%, and for urine 12.2% in 10 analyses of the same plasma and urine with the protein-precipitation procedure, as compared with 26.0 and 38.2%, respectively, by the acid-digestion technique.
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Medicated feeds (pelleted or mash) containing guarantees of carbadox, furazolidone, nitrofurazone, and ethopabate are pretreated with water, extracted with 95% dimethylformamide overnight at room temperature, cleaned up on a column of alumina, and injected into a high pressure liquid chromatograph for quantitative measurement. Carbadox, nitrofurazone, and furazolidone can be separated; chromatograms show excellent baseline resolution, and results are in good agreement with colorimetric methods. The same extraction and cleanup can be used to improve colorimetric methods for furazolidone and nitrofurazone.
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