Deproteinization is Necessary for the Accurate Determination of Ammonia Levels by Glutamate Dehydrogenase Assay in Blood Plasma From Subjects With Liver Injury
Language English Country England, Great Britain Media print
Document type Journal Article
PubMed
29126300
DOI
10.1093/labmed/lmx053
PII: 4372133
Knihovny.cz E-resources
- Keywords
- NADH oxidation, NADPH oxidation, ammonia, glutamate dehydrogenase, liver enzymes, liver injury,
- MeSH
- Ammonia blood MeSH
- Blood Chemical Analysis methods standards MeSH
- Glutamate Dehydrogenase metabolism MeSH
- Blood Proteins isolation & purification MeSH
- Rats MeSH
- Humans MeSH
- Linear Models MeSH
- NAD metabolism MeSH
- NADP metabolism MeSH
- Liver Diseases blood metabolism MeSH
- Oxidation-Reduction MeSH
- Rats, Wistar MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Humans MeSH
- Male MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Ammonia MeSH
- Glutamate Dehydrogenase MeSH
- Blood Proteins MeSH
- NAD MeSH
- NADP MeSH
OBJECTIVE: To determine the effect of presence of high concentrations of nicotinamide adenine dinucleotide (NADH)- and nicotinamide adenine dinucleotide phosphate (NADPH)-consuming enzymes on the accuracy of glutamate dehydrogenase (GLDH) assay for ammonia. METHODS: We measured ammonia concentrations using GLDH and NADH or NADPH in blood-plasma specimens and specimens deproteinized by sulfosalicylic acid from CCl4-treated or control rats. The nonspecific oxidation of NADH and NADPH was measured in mixtures without GLDH. RESULTS: We observed a gradual decrease (~0.5%) in absorbance in the plasma of controls after the addition of NADH but not after adding NADPH. The decrease in absorbance in plasma of CCl4-treated animals was 13.2% and 5.2% after the addition of NADH and NADPH, respectively. The decrease in absorbance was not detected in deproteinized specimens. The values of ammonia concentration were higher in the plasma specimens compared with the deproteinized ones. CONCLUSION: Deproteinization is necessary for accurate measurement of ammonia using GLDH assay in the blood plasma of subjects with liver injury.
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