Uric acid

Introduction
The uric acid is the end product of purine
metabolism.
The purines adenine and guanine are
constituents of both types of nucleic acid (DNA &
RNA).
Uric acid is the end product of nuclear
materials.
(1)Diet (food containing spleen , liver , red meat)
rich in nuclear materials.
(2)Tissues contain DNA , RNA (Catabolism).
(3)Free nucleotides (ATP ,GTP , AMP).

Sources of purines

(4)Hypoxanthine and Xanthine.

Uric acid is synthesized in the liver, in a reaction

requiring the enzyme xanthine oxidase.
From the liver , uric acid is transported to the
kidneys where it is filtered by the glomeruli.
Nearly all the filtered uric acid is reabsorbed in
the proximal tubules ; and small amounts are
then secreted by the distal tubules and ultimately
appear in the urine.
Nearly all of the uric acid in the plasma is in the
form of monosodium urate.
Urates in this form are relatively insoluble.
At high levels (>6.4mg/dl) , the plasma is
saturated ; urate crystals may form and
precipitate in the tissues.

Clinical significant of uric

acidranges
Normal
Adult male:-2-7.2mg/dl.
Adult female:-0.5-6mg/dl.

Hyperuricaemia
Primary

Secondar
y

(A)Primary Hyperuricaemia
(1)Chronic renal failure.
(2)Gout.
(2)Increase breakdown of cells as in:(a)Leukaemia.
(b)Treatment of malignancy (Chemotherapy &
(B)Secondary Hyperuricaemia
radiotherapy).
(1)Polycythaemia.
(2)Acidosis.
(3)Pneumonia.
(4)Toxins.
(5)Drugs.
(6)Post-menopausal women.

Complication of hyperuricaemia
Serious consequences of high plasma uric acid
is due to the insolubility of uric acid and sodium
mono-urate and precipitated in the:-

(A)Kidney
Caused:-(1)Renal stones.
(2)Renal failure.

(B)Cartilage and around the joints

Causing arthritis.

Hypouricaemia
(1)Liver disease.
(2)Defective tubular reabsorption (Fanconi
syndrome).
(3)Chemotherapy with azathioprine or 6mercaptopurine.
(4)Overtreatment with allopurinol.

Methods used for

estimation of uric acid

(1)Phosphotungestic acid
method
Principle
Phosphotungstic acid is reduced by uric acid in
the presence of sodium cyanide or carbonate to
give blue complex which can be measured
colorimetrically.
Carbonate has advantages over cyanide:(1)Gives lower blanks.
(2)More stable colours.
(3)More reproducible absorbances from batch to
batch.
(4)It is not poisonous.
It has the disadvantages of being less sensitive.

Urea was added to the cyanide in an attempt to

prevent this ; however , more recently , the
addition of lithium in the reaction mixture
together with the use of a reliable commercially
prepared phosphotungstate reagent have almost
brought it under control.
Serum should preferably be stored overnight at
4C before analysis ; this will eliminate the
possible error due to ascorbic acid , which is
probably the main , but fortunately very labile ,
non-uric acid reducing substance contributing in
this reaction.
Therapeutic levels of paracetamol have also
been found to interfere with phosphotungstate

(2)Enzymatic methods
(uricase method)
(A)End point method
Principle
Enzyme uricase converts uric acid to allantonin
and hydrogen peroxide.
In the presence of peroxidase , hydrogen
peroxide reacts with phenolic chromogens to form
red coloured compound.
The intensity of red colour is proportional to the
amount of uric acid in the sample.

Uric acid H 2O O 2

Allantonin H 2O 2

H 2 O Phenolic chromogens

Uricase

Red colour compound

peroxidase

(B)Kinetic method
Uric acid H 2 O O 2

Allantonin
Uricase

The allontonin is flow-up (monitored)

spectrophotometrically at 293nm.
The amount of which is proportional to the
concentration of uric acid in the sample.