Abstract
Determination of ammonia level in blood is important, especially in the diagnosis of hepatic disorders. An indigenously purified enzyme was used in the standardisation of the assay. The assay is a two reagent system, requires five minutes for completion and can be performed at temperature between 25–27°C. Performance of the assay was assessed by linearity, imprecision, functional sensitivity and interference studies. Lyophilised reagent I and reagent II were found stable for at least one year. The plasma level of ammonia for the controls was 13.7±7.3 μMol/L, whereas for subjects of hepatic disorders, it was 69.1±32.4 μMol/L (P<0.001). The functional sensitivity was between 2–1000 μMol/L. Within-run coefficient of variation was between 1.1–2.0% and between-run coefficient of variation was between 1.9–3.7%. The mean recovery after dilution was 99.6%. The present method can estimate ammonia up to 1000 μMol/L without dilution of sample. Assay time of five minute may be shortened to one minute. This method is suited for routine clinical use in treatment of hepatic disorders.
Key Words: Ammonia, enzymatic assay, glutamate dehydrogenase, deaminase inhibitor
Full Text
The Full Text of this article is available as a PDF (467.2 KB).
References
- 1.Ballistreri W.F., Rej R. Liver Function. In: Burtis C.A., Ashwood E.R., editors. Tietz Textbook of Clinical Chemistry. 2nd edition. Philadelphia, USA: WB Saunders; 1994. pp. 1449–1513. [Google Scholar]
- 2.Diaz J., Tornel P.L., Martinez P. Reference intervals for blood ammonia in healthy subjects, determined by microdiffusion. Clin. Chem. 1974;41:1048–1048. [PubMed] [Google Scholar]
- 3.Cabral Joao P.S. Comparison of methods to assay ammonia in bacterial suspensions. J. Microbiol. Methods. 1974;19:207–213. doi: 10.1016/0167-7012(94)90071-X. [DOI] [Google Scholar]
- 4.Huizenga J.R., Tangerman A., Gips C.H. Determination of ammonia in biological fluids. Ann. Clin. Biochem. 1994;31:529–543. doi: 10.1177/000456329403100602. [DOI] [PubMed] [Google Scholar]
- 5.Ratcliff C.R., Hall F.F. Ammonia in plasma, enzymatic procedure. Sel. Methods. Clin. Chem. 1982;9:85–90. [Google Scholar]
- 6.Anken H.C., Schiphorst M.E. Kinetic determination of ammonia in plasma. Clin. Chim. Acta. 1974;56:151–157. doi: 10.1016/0009-8981(74)90223-X. [DOI] [PubMed] [Google Scholar]
- 7.Fonseca-Wollheim F., Heinze K.G. Which is the appropriate coenzyme for measurement of ammonia with glutamate dehydrogenase? Eur. J. Clin. Chem. Clin. Biochem. 1992;30:537–540. [PubMed] [Google Scholar]
- 8.Pesh-Imam M., Kumar S., Willis C.E. Enzymatic determination of plasma ammonia: evaluation of Sigma and BMC kits. Clin. Chem. 1978;24:2044–2046. [PubMed] [Google Scholar]
- 9.Kurahasi K., Shihara A., Uehara H. Determination of ammonia in blood plasma by ion exchange method. Clin. Chim. Acta. 1972;42:141–146. doi: 10.1016/0009-8981(72)90387-7. [DOI] [PubMed] [Google Scholar]
- 10.Solberg H.E. Establishment and use of reference values. In: Burtis C.A., Ashwood E.R., editors. Tietz Textbook of Clinical Chemistry. 2nd edition. Philadelphia, USA: WB Saunders; 1994. pp. 454–484. [Google Scholar]
- 11.Howanowitz J.H., Howanowitz P.J., Skrodzki C.A., Iwanski J.A. Influence of specimen processing and storage conditions on results of plasma ammonia. Clin. Chem. 1984;30:906–908. [PubMed] [Google Scholar]
- 12.Linder A., Bauer S. Effect of temperature, duration of storage and sampling procedure on ammonia concentration in equine blood plasma. Eur. J. Clin. Biochem. 1993;31:473–476. doi: 10.1515/cclm.1993.31.7.473. [DOI] [PubMed] [Google Scholar]
