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Uric Acid
Uric acid is a metabolite of nucleic acids, purines, and nucleoproteins and is the end product of
protein (purine) metabolism in man. In most mammals, uric acid is oxidized in the liver to
allantoin. Higher apes and man appear incapable of destroying uric acid efficiently in this
manner. As a result, uric acid accumulates in the plasma (as sodium urate) and is excreted in
the urine. Occasionally some persons are afflicted with extensive deposits of uric acid (urates)
in their tissues, causing a disorder termed gout. Increased levels of uric acid have been
observed in renal failure, chronic lead poisoning, polycythemia, some leukemias, and toxemia
of pregnancy.
Reaction
Uricase >
Uric acid + 2H2 O + O2 Allantoin + CO2 + H2 O2
Peroxidase >
H2 O2 + 4 aminophenazone + DCHB N-(4-antipyryl)-3-chloro-5-sulfonate-
p-benzo-quinonemoine Dye + H2 O
(absorbance maximum at 520 nm)
Supplies and Equipment
Reagents
1. Uricase – enzyme prepared from Candida utilis.
2. Horseradish Peroxidase
3. 4-Aminoantipyrine
4. 3,5-Dichloro-2hydroxybenzenesulfonic acid
5. Phosphate Buffer, pH 7.5
6. Uric Acid Standard Solution, 8mg/dL Cat. No. 1044 - an aqueous solution of uric
acid with solubilizer and stablizer added.
*DANGER / PRECAUTIONS:
1. Uric acid reagents are for in-vitro diagnostic use only. Normal precautions should be
used as for handling laboratory reagents, and all federal, state, and local laws for
handling and disposal followed.
2. Avoid contact and inhalation
3. Consider all blood derivatives as potentially hazardous.
Specimens
1. Serum or plasma (heparinized or EDTA). UA is stable for 2-3 days in serum at room
temperature, 3-7days at 2-8°C, and 6-12 months frozen. Slight hemolysis has no effect
upon UA level.
Interfering substances
Steps
NOTE: Procedure can be performed on 24 hour urine collections. Measure and record urine
volume. Mix well. A 10-fold dilution (1 + 9) of the urine sample is required for the
procedure. Once the uric acid concentration of the dilution is determined, multiply
by the x 10 dilution factor.
Calculation (Serum)
1. Calculate control and patient results using the following formula.
(1)
Quality Control
Two levels of assayed serum controls.
Linearity
When performed as directed, procedure is linear to 20 mg/dL
Expected Values (It is recommended that each laboratory establish a normal range.)
Serum Male 3.6 - 7.7 mg/dL
Female 2.5 - 6.8 mg/dL
Urine 250-750 mg/24 hr (dependent on diet content of purines)
Name
Date
Uric Acid
Wavelength _____________
Linearity _____________ Spectrophotometer Used ______________________
Identification Absorbance Concentration (units)
Standard _____________________
Standard _____________________
Standard _____________________
Control 1 _____________________
Control 2 _____________________
Level 1 ID______________
Level 2ID_______________