Uricase- version 1

UNIT DEFINITION

One unit (U) is defined as the amount of enzyme which consumes 1 μmol of uric acid per min under the conditions described below.

REAGENTS PREPARATION

Reagent I: 0.001% uric acid.

Enzyme diluent: Add 3.09 g boric acid, 0.37 g EDTA-Na2·2H2O and 0.01 g Triton X-100 in 800 ml pure water, adjust to pH 8.5 by NaOH and set to 1000 mL by pure water.

Sample: The enzyme was diluted to 0.05-0.1 U/mL with enzyme diluent.

PROCEDURE

1. Preincubate the Reagent I at 37 ℃.
2. Add 2 mL of Reagent I, 0.5 mL of pure water, and then add the diluted enzyme to a cuvette.

3. Record the ΔAs at 290 nm in 1 minute in a spectrophotometer thermo stated at 37 ℃.

At the same time, measure the blank rate ΔAb by using the same method as the test except that the enzyme diluent is added instead of the enzyme solution.

∆A = ∆As – ∆Ab