Name: Medical Biochemistry 2

Group: Date:

DETERMINATION OF URIC ACID IN SERUM

Principle
Uric acid (2,6,8-trihydroxypurine) reduces a phosphotungstate reagent (containing W+VI) to blue
compound in the presence of Na2CO3. Determination is made in deproteinated serum and the density
of blue color is measured photometrically at wavelength of 610 nm.

Reagents
• Serum
• Standard solution of uric acid 200 µmol/l
• Phosphotungstate reagent
• Deproteinating reagent (1.2 mol/l trichloroacetic acid, TCA)
• Na2CO3 0.1 mol/l
• Distilled water

Procedure
Prepare sample, standard and blank solution in centrifugation test tubes according to Table:

Sample (ml) Standard (ml) Blank (ml)

Serum 1 - -
Standard - 1 -
Distilled water 4 4 5
TCA 1 1 1
Mix and incubate 10 min at laboratory temperature. Sample centrifugation for 10
min at 300g at 20°C. Then pipette into glass test tubes
Supernatant 2 2 2
Phosphotungstate 0.1 0.1 0.1
reagent
Na2CO3 0.9 0.9 0.9
Mix and incubate 10 min at laboratory temperature

Measure the absorbances of sample and standard against blank at wavelength of 610 nm.

Results
Asample =
Astandard =
Name: Medical Biochemistry 2
Group: Date:
Calculation
Calculate the concentration of uric acid according to formula:

𝐴𝑠𝑎𝑚𝑝𝑙𝑒
Uric acid (μmol/l) = × 𝑐𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑
𝐴𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑

Reference values
Men: 208 - 428 μmol/l
Women: 155 - 357 μmol/l

Conclusion