Kinetic assay of serum and urine for urea with use of urease and leucine dehydrogenase
1997; American Association for Clinical Chemistry; Volume: 43; Issue: 10 Linguagem: Inglês
10.1093/clinchem/43.10.1932
ISSN1530-8561
AutoresYoshitaka Morishita, Kiyoshi Nakane, Toshiaki Fukatsu, Nobuo Nakashima, Katsumi Tsuji, Yoshihiro Soya, Keizo Yoneda, Shigeki Asano, Yoshihisa Kawamura,
Tópico(s)Hyperglycemia and glycemic control in critically ill and hospitalized patients
ResumoAbstract 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 itsKmvalue for NH4+ at pH 8.75 is large (∼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, Sy|x = 0.34 mmol/L, n = 100) for sera, and y = 1.070x − 12.6 mmol/L (r = 0.998, Sy|x = 7.41 mmol/L, n = 100) for urine.
Referência(s)