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Uric Acid Test Procedure

Uric acid is the end product of purine metabolism. Uric acid is excreted to a large degree by the kidneys and to a smaller degree in the intestinal tract by microbial degradation. Increased levels are found in Gout, arthritis, impaired renal functions.and starvation. Decreased levels are found in Wilson’s disease, Fanconis syndrome and yellow atrophy of the liver.

Uric Acid Test Procedure
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Principle:

Uricase converts uric acid to allantoin and hydrogen peroxide. The hydrogen peroxide formed further reacts with a phenolic compound and 4 aminoantipyrine by the catalytic action of peroxidase to form a red coloured quinoneimine dye complex. Intensity of the colour formed is directly proportional to the amount of uric acid present in the sample.

Uric Acid Principle

Normal reference values

Normal Values mg/dl
Male3.4 – 7.0 mg/dl
Female2.5 – 6.0 mg/dl
Normal Values

Contents:

ContentVolume
Pack Sizes25 ml, 75 ml, 2 x 75 ml, 2 x 150 ml
L1 : Buffer Reagent20 ml, 60 ml, 2 x 60 ml, 2 x 120 ml
L2 : Enzyme Reagent5 ml, 15 ml, 2 x 15 ml, 2 x 30 ml
S : Uric Acid Standard (8 mg/di)5 ml, 5 ml, 5 ml, 5 ml
Kit Components

Reagent Preparation:

All Reagents are ready to use.

Reagents Storage/ stability:

Contents are stable at 2-8°C till the expiry mentioned on the labels.

Sample :

Serum, plasma. Uric Acid is reported to be stable in the sample for 3 -5 days when stored at 2-8°C.

System Parameters:

ReactionEnd Point
Wavelength520 nm (Hg 546 nm)
FilterYellow Green
Zero SettingReagent Blank
lncub. Temp.37°C I R.T
lncub. Time5 min./ 15 min.
Delay Time—-
Read Time—-
No. of read.—-
Interval—-
Sample Vol.0.02 ml
Reagent Vol.1.00 ml
Standard8 mg/di
Factor—-
React. SlopeIncreasing
Linearity20 mg/di
Unitsmg/di
Uric Acid System Parameters

Test Procedure:

Pipette into clean dry test tubes labelled as Blank (B), Standard (S), and Test (T):

Addition SequenceBlank (B)Standard (S)Test (T)
L1 : Buffer Reagent800 μl800 μl800 μl
L2 : Enzyme Reagent200 μl200 μl200 μl
Distelled Water20 μl
Standard20 μl
Sample20 μl

Mix well and incubate at 37°C for 5 min. or at R.T. (25°C) for 15 min. Measure the absorbance of the Standard (Abs.S), and Test Sample (Abs.T) against the Blank, within 30 Min.

Calculations:

Uric Acid Test Calculations

Linearity:

This procedure is linear upto 20 mg/di. If values exceed this limit, dilute the serum with normal saline (NaCl 0.9%) and repeat the assay. Calculate the value using the proper dilution factor.

Related Articles:



Possible References Used

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