New enzymatic assay with urea amidolyase for determining potassium in serum.
We developed a new simple assay for potassium ion in serum using urea amidolyase (UAL) from yeast sp. The method is based on activation of the enzyme by potassium ion. We eliminated endogenous ammonium ion by use of glutamate dehydrogenase (GLDH), and then monitored the production of ammonium ion by UAL, urea, ATP, bicarbonate and magnesium ions. Ammonium ion was produced proportional to the potassium ion concentration and was determined by adding GLDH to produce NADP+ in the presence of 2-oxoglutarate and NADPH. We monitored the change of absorbance at 340 nm. The inhibitory effect of calcium ion to this assay was eliminated by adding glycoletherdiamine-N, N, N', N'-tetraacetic acid to the reaction. The within-assay coefficients of variation (CV) of this method were 0.9-1.55% (n = 10) at 3.32-6.18 mmol/L. Day-to-day CVs ranged from 1.49% to 2.46%. The analytical recovery was 96-108%. The correlation coefficient between the values obtained by our method (y) and those by the ion-selective electrode (ISE) method (x) was 0.994 (y = 1.032x-0.166 mmol/L, Syx = 0.110, n = 100). The presence of bilirubin, haemoglobin or other ions did not affect this assay, confirming the usefulness of this assay for clinical purposes.
Kimura
S
S
Central Laboratory for Clinical Investigation, Osaka University Hospital, Japan.
Iyama
S
S
Yamaguchi
Y
Y
Hayashi
S
S
Fushimi
R
R
Amino
N
N
D000085:Acetates; D000255:Adenosine Triphosphate / Q000378:metabolism; D001774:Blood Chemical Analysis / Q000379:methods*; D002118:Calcium / Q000097:blood; D002138:Calibration; D019731:Carbon-Nitrogen Ligases; D002614:Chelating Agents; D004789:Enzyme Activation; D005969:Glutamate Dehydrogenase; D006801:Humans; D006863:Hydrogen-Ion Concentration; D017736:Ion-Selective Electrodes; D007700:Kinetics; D008025:Ligases / Q000378:metabolism*; D011188:Potassium / Q000097:blood*; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D012680:Sensitivity and Specificity; D013053:Spectrophotometry
D000085:Acetates; D002614:Chelating Agents; D000644:Quaternary Ammonium Compounds; D000255:Adenosine Triphosphate; D005969:Glutamate Dehydrogenase; D008025:Ligases; D019731:Carbon-Nitrogen Ligases; C023270:urea carboxylase (hydrolyzing); D011188:Potassium; D002118:Calcium