Comparative In vitro Hipouricemia Activity of

Sida rhombifolia Linn Leaves Extracts used

Xanthine Oxidase Enzyme



Background: Sidaguri plants traditionally used as urolithiasis or for treating kidney stones.
Generally, kidney stones are calcium oxalate stones, but can also come from a mixture of uric
acid and calcium oxalate.
Objective: To know the characteristics of sidaguris leaves and to determine the effects of
sidaguris leaves to decrease of uric acid.
Urikostatic drug class inhibits the enzyme xanthine oxidase which converts hypoxanthine
into xanthine and xanthine into uric acid. Thus the production of uric acid and xanthin
production decreases and increases hipoxanthin. Examples of the medicine is Allopurinol.
Plant sidaguri (Sida rhombifolia L) has been used empirically as a natural medicine by the
people in the treatment of gout. Flavonoids contained from sidaguri leaf extract in vitro inhibitor
of xanthine oxidase have effect so as to reduce the production of excess uric acid.

Extraction (Maceration Methods)
500 gram of dried Sidaguri leaves (crushed/cut small/moderately coarse powder)

Placed in closed vessel

Etanol 70 % solvent (menstruum) added

Allowed to stand for 3x24 hours

Liquid strained of

Solid residues (mark) pressed (recover as much as occluded solution)

(Strained and expressed liquids mixed)

Clarified by subsidence or filtration

Evaporation and concentration


Principle: the enzyme xanthine oxidase catalyzes the oxidation of hypoxanthine and xanthine
into uric acid that plays a role in the formation of uric acid that causes gout. During reoxidation
xanthine oxidase, molecular oxygen acts as an electron acceptor, generating superoxide and
hydrogen peroxide. The reaction is as follows:
Xanthine + 2 H
2
O Gout + 2 O
2
+ 2 H
+

Xanthine + 2 O
2
+ 2 H
2
O Gout + H
2
O
2

In vitro Testing Procedure

A total of 1 mL of sample extract with various concentrations prepared

added 3 mL of 0.05 M phosphate buffer pH 7.5 and 2 mL of xanthine substrate solution

incubated for 10 min at 37 C

added 0.1 mL of xanthine oxidase

incubated for 30 min at 37 C

The reaction was stopped with adding 6 ml of HCl 1 N

Testing blanks as much as 3 mL phosphate buffer and 2 mL of xanthine substrate solution mixed
in the vial then incubation respectively at 37 C for 10 minutes

followed by the addition of 0.1 mL of xanthine oxidase

The solution was then homogenized using a vortex mixer, incubated at 37 C for 30 minutes

The reaction was stopped by adding 1 mL of 1 N HCl

solution was measured at a wavelength of 291 nm with a spectrophotometer


Dried sampel weight : 500 Gr
Cup (without handle) empty weight : 72,81 Gr
Cup with extract : ?
Solvent used : 2,8 L
Organoleptic of viscous extract : brown, bitter taste, smell like jamu