A rapid method for plasma ammonia estimation using an indigenously purified enzyme

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.