title: Kinetic assay of serum and urine for urea with use of urease and leucine dehydrogenase.
abstract: We describe a new kinetic assay for determining urea in serum or urine with use of urease (EC 3.5.1.5) and leucine dehydrogenase (EC 1.4.1.9). The latter enzyme is suitable for the kinetic assay of NH4+ because its Km value for NH4+ at pH 8.75 is large (approximately 500 mmol/L). Interference from endogenous NH4+ in serum or urine is obviated by subtraction of the assayed endogenous NH4+ value in a sample blank. For serum, within-assay CVs (n = 10) were 0.39-0.58%; day-to-day CVs (n = 10) were 1.56-2.30%. In urine, within-assay CVs (n = 10) were 0.86-1.15%. Analytical recovery of urea (0.893-71.4 mmol/L) added to patients' sera (urea 6.14 mmol/L) was 99.2-105.2%. The calibration curve for serum was linear through zero for urea concentrations up to 142.9 mmol/L and for urine up to 714.3 mmol/L. No influences of added ammonium ion, bilirubin, hemoglobin, ascorbic acid, or Intralipid were observed. The regression equations for this method (y) and conventional methods (x = Determiner-LUN for serum assays, Serotec UUR-R for urine) were: y = 1.016x - 0.12 mmol/L (r = 0.999, S(y/x) = 0.34 mmol/L, n = 100) for sera, and y = 1.070x - 12.6 mmol/L (r = 0.998, S(y/x) = 7.41 mmol/L, n = 100) for urine.
authors.affiliation: Department of Clinical Laboratory, Nagoya University Hospital, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Morishita
authors.forename: K
authors.initials: K
authors.lastname: Nakane
authors.forename: T
authors.initials: T
authors.lastname: Fukatsu
authors.forename: N
authors.initials: N
authors.lastname: Nakashima
authors.forename: K
authors.initials: K
authors.lastname: Tsuji
authors.forename: Y
authors.initials: Y
authors.lastname: Soya
authors.forename: K
authors.initials: K
authors.lastname: Yoneda
authors.forename: S
authors.initials: S
authors.lastname: Asano
authors.forename: Y
authors.initials: Y
authors.lastname: Kawamura
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000378:metabolism*; D016477:Artifacts; D001806:Blood Urea Nitrogen; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D050803:Leucine Dehydrogenase; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014510:Urease / Q000378:metabolism*
chemical_list: D000594:Amino Acid Oxidoreductases; D050803:Leucine Dehydrogenase; D014510:Urease