ANTIHYPERURICEMIC EFFECT OF AJWA DATE FRUIT (Phoenix

dactylifera L.) EXTRACT IN POTASSIUM OXONATE-INDUCED

HYPERURICEMIC MICE
Veramita Augusta Arisandy1, Tsamara Nurwina Nugroho1, Sanubari Kukuh Samahita1, Ainun Rahmasari
Gumay2*
1
Medical Student, Faculty of Medicine, Diponegoro University, Semarang, Indonesia
2
Lecturer, Department of Physiology, Faculty of Medicine, Diponegoro University, Semarang, Indonesia

*)
Corresponding author: Ainun Rahmasari Gumay. Address: Faculty of Medicine Diponegoro University, Jl. Prof.
Soedharto SH Tembalang Semarang, Central Java, Indonesia, 50275. Email: ainungumay@fk.undip.ac.id

ABSTRACT

Introduction. Hyperuricemia can lead to serious complication such as cardiovascular disease,

which is the first cause of death in the world. Ajwa date fruit (Phoenix dactylifera L.) have been
shown to contain high flavonoid content. It has the ability to reduce the formation of uric acid by
inhibiting xanthine oxidase enzyme. This study aims to prove the effect of Ajwa date fruit extract
on uric acid formation in potassium oxonate-induced hyperuricemic mice.
Method. An experimental study with post-test only controlled group design. The samples were
30 male Balb/c mice, divided into 6 groups. Group C1 was normal control group only treated
with aquadest. Group C2 was induced by potassium oxonate and treated with 10 mg/kgBW of
allopurinol. Group C3 were induced by single intraperitoneally injection of 250 mg/kgBW of
potassium oxonate. Group AE 1, 2, and 3, each was induced by potassium oxonate and orally
administered by Ajwa date fruit extract with the dose 3,64; 7,28; 14,56 mg/kgBW. Blood
samples in each group will be collected in 1 hour after giving the treatment. Uric acid level will
be measured using uric acid FS* TBHBA reagent.
Results. The uric acid level in AE1, AE2, ad AE2 group was lower than C3 group. The uric acid
level in C2 group was lower than AE1, AE2, and AE3, and C2 group. The uric acid level in AE1
group was significantly lower than AE2 and AE3 group.
Conclusion. Ajwa date fruit extract may decrease the uric acid level in potassium oxonate-
induced hyperuricemic mice.

Keywords : Ajwa date fruit, hyperuricemia, potassium oxonate


INTRODUCTION
Gout arthritis is a disease characterized by
gout and accumulation of urate crystals in
the joints.(1) According to the WHO in
2013, as many as 81% of gout patients in
Indonesia, only 24% went to the doctor,
while 71% used pain reliever. Based on
2013 data from Riset Kesehatan Dasar
(Riskesdas), 11.9% of diagnoses of gout
were obtained from health workers, while
24.7% were obtained from the results of
the research based on detected symptoms.
Arthritis is caused by high uric
acid levels and followed by pain and
swelling in the joints. In addition, high
uric acid levels can cause complications
such as cardiovascular disease, the
number-one cause of death in the world
(WHO, 2016). This is related to the
formation of atherosclerotic plaques
which can be triggered by the process of
gout formation. In this process,
hypoxanthine will be converted to
xanthine, which then later becomes uric
acid with the help of xanthine oxidase and
produce reactive oxygen that has a
stimulating effect on the walls of blood
vessels so that it becomes a factor that
causes atherosclerosis.(2)
Uric acid presents in normal
condition of the body, but excessive
amount of it can cause accumulation of
crystals in the joints and become a
disease. Increased uric acid levels beyond
normal range are called hyperuricemia.
Hyperuricemia occurs when uric acid
levels in the serum are >7.0 mg / dl in
men and >6.0 mg / dl in women.(3) In the
case of hyperuricemia, reactive oxygen
will be produced and it will increase the
atherosclerotic plaque formation.
Ajwa dates or commonly known
as Prophet’s dates are the types of dates
that grow in Saudi Arabia. The Prophet
2
(peace and blessings of Allah be upon
him) said: “Whoever eats seven Ajwa
dates in the morning, will not be harmed
by any poison or witchcraft that day.“
(Muttafaqun‘alaih. Bukhari 5779 and Mus
lim no.2047). Ajwa dates extracts have the
highest concentration of polyphenols
compared to other types.(4) One
polyphenol element in Ajwa dates are
flavonoid.(5) Flavonoid has the ability to
inhibit xanthine oxidase enzyme, thus
reducing the formation of uric acid.(6)
In the present times, allopurinol is
the standard therapy in gout during the
period between acute attacks, which
reduces total uric acid in the body by
inhibiting xanthine oxidase. However,
allopurinol has several side effects such as
nausea, vomiting, diarrhea, bone marrow
suppression, and allergic skin reactions.(7)
This study aims to utilize the potential of
Ajwa dates in reducing uric acid levels, so
that it can help reduce the side effects
resulting from uric acid-lowering drugs.
METHOD
Ajwa Dates Extraction
500 g of Ajwa dates was dried in an oven
at 80oC for 48 hours. Dried Ajwa dates
were mashed by using blender, then
macerated using ethanol 70% with a ratio
of 1:2 for 48 hours and stirred for the first
24-hour.(8) Afterwards, the filtrate was
evaporated using rotatory evaporator at
±75oC until dense filtrate obtained.
Treatment of Potassium Oxonate-
Induced Hyperuricemic Mice
The study conducted for three weeks and
carried out at the Animal Laboratory and
Biology Laboratory, Faculty of
Mathematics and Natural Sciences,
Universitas Negeri Semarang (UNNES).
This research was an experimental study

with a posttest only control group design
and used animals as research objects.
This study used 2-3-month-old
male Balb/C mice divided into 6 groups
with 5 mice in each group. One hour
before giving the treatment, group C2, C3,
AE1, AE2, and AE3 were induced by
single intraperitoneally injection of 250
mg/kgBW of potassium oxonate Group
C1 or normal control group only treated
with aquadest. Group C2 was treated with
10 mg/kgBW of allopurinol. Group C3
was only induced by potassium oxonate.
Group AE 1, 2, and 3, each was orally
administered by Ajwa date fruit extract
with the dose 3,64; 7,28; 14,56
mg/kgBW.(9)
Uric Acid Level Measurement
One hour after giving the
treatment, blood samples were taken via
ophthalmicus vein from each group. The
blood samples were centrifuged to get its
serum. Then, uric acid levels were
measured using uric acid FS* TBHBA
reagent.
RESULTS
The Effects of Giving Potassium
Oxonate in Enhancing Uric Acid Levels
The research data on the effect of giving
Ajwa date fruit extract to mice that have
been obtained was analyzed using One
Way ANOVA method on computerized
statistical program. This experiment was
initiated by inducing potassium oxonate in
3
several groups of mice to increase uric
acid levels. Subjects in the normal group
(group C1) were not induced with
hyperuricemia but were only given
aquadest. The subjects in the positive
control group (group C2) was induced by
potassium oxonate and treated with 10
mg/kgBW of allopurinol, the negative
control group (group C3) was induced by
single intraperitoneally injection of 250
mg/kgBW of potassium oxonate, the
treatment groups (group AE1, AE2, and
AE3) were induced with hyperuricemia
via intraperitoneal potassium oxonate
administration with the dose of 250
mg/kgBW.
Statistical test showed that the uric
acid levels in group C1 and group C3
showed a significant difference in uric
acid levels with a significance value of
0.000 (p <0.05). Group C1 and group C2
showed an insignificant difference in uric
acid levels with a significance value of
0.157 (p> 0.05). Whereas, group C2 and
group C3 showed a significant difference
in uric acid levels with a significance
value of 0.000 (p <0.05). This shows that
the administration of allopurinol at a dose
of 10 mg/kgBW in group C2 gave a
significant decrease in uric acid levels
compared to group C3 induced by
hyperuricemia without allopurinol
administration. Comparisons of uric acid
levels in each group are shown in Graph
1.
 4

Graph 1. Comparison of Mean of Uric Acid Levels of Subject in Each Group

Effects of Giving Ajwa Dates Extract insignificant result with a significance

(Phoenix dactylifera L.) value of 0.103 (p> 0.05).
The administration of Ajwa date fruit The mean of uric acid levels in
extract at a dose of 3.64 mg/kgBW (group each treatment group (group AE1, AE2,
AE1) showed an insignificant result when and AE3) are shown in Table 1.
compared to group C1 with a significance
value of 0.335 (p> 0.05). Whereas,
compared to group C2 and group C3, it
showed significant results of uric acid
level differences with significance values
of 0.024 and 0.000 consecutively (p<0,05).
The administration of Ajwa date
fruit extract at a dose of 7.28 mg/kgBW
(group AE2) showed significant results
compared to group C1, C2, and C3 with a
significance values of 0.013, 0.001, and
0.013 consecutively (p <0.05).
The administration of Ajwa date
fruit extract with a dose of 14.56
mg/kgBW (group AE3) showed significant
results compared to groups C1 and C2
with significance values of 0.001 and
0.000 consecutively (p <0.05). Whereas,
compared to group C3, it showed an

Table 1. Mean of uric acid levels in each groups
No. Groups
1. Normal (C1)
2. Positive control (C2)
3. Negative control (C3)
4. Ajwa Date Fruit Extract 3,64 mg/kgBW (AE1)
5. Ajwa Date Fruit Extract 7,28 mg/kgBW (AE2)
6. Ajwa Date Fruit Extract 14,56 mg/kgBW (AE3)
Based on the table above, it was found that
the administration of Ajwa dates extract of
3.64 mg/kgBB (group AE1) showed the
largest decrease in uric acid levels compared
DISCUSSIONS
In the positive control group (group C2), one
hour after potassium oxonate was induced,
allopurinol was given 10 mg/kgBW and
showed a significant effect compared to
group C2. This is in accordance with the
benefit of allopurinol which is used as a
therapy to reduce uric acid levels by
inhibiting the formation of purine through
the action of inhibited xanthine oxidase
enzymes.(10) Inhibition of xanthine oxidase
causes hypoxanthine and xanthine to be
excreted in urine so that uric acid levels in
the blood and urine decreases.(11)
In the negative control group (C3
group), which was induced hyperuricemia
by administering potassium oxonate at a
dose of 250 mg/kgBW, it showed higher
uric acid levels compared to other groups.
This is consistent with the statement that
potassium oxonate is used as an induction of
hyperuricemia because it is a competitive
uricase inhibitor to increase uric acid levels
5
Uric Acid Level (mg/dL)
0.8200±0.90233
0.3025±0.14891
2.7525±0.01702
1.1675±0.29489
1.7825±0.31904
2.1500±0.14018
to group AE2 and group AE3 with an
average value of uric acid levels of 1.1675
mg/dL.
by preventing changes in uric acid to
allantoin.(12)
In group AE1 given Ajwa date fruit
extract 3.64 mg/kgBW showed a decrease in
uric acid levels greater than uric acid levels
produced in group AE2 (7.28 mg/kgBB) and
group AE3 (14.56 mg / kgBB). Meanwhile,
the highest dose of extract extract in this
study, namely the dose of 14.56 mg/kgBB
(group AE3) showed a lower decrease in
uric acid levels compared to groups AE1
and AE2. In this case, the increase in drug
dosage should be proportional to the
increase in response, however, by increasing
doses, the response will eventually decrease
because a maximum dose has been reached
which no longer can increase response.(13)
This often occurs in natural materials
because the components of the compounds
they contain consist of several chemical
compounds that can interact to cause certain
effects. With the increase in dosage, the
number of components of chemical

compounds contained increased, so that
adverse interactions can occur and cause a
decrease in the effect of substances. The
limited number of receptors can also reduce
the effect because not all drugs can bind to
the receptor, so even if the dose is increased,
the response will not increase.(13) Ajwa
date fruit extract also contains flavonoids
which can inhibit the action of the xanthine
oxidase enzyme in the formation of uric
acid, so that uric acid levels in mice can
decrease.(6)
REFERENCES
1. Saigal R, Agrawal A. Pathogenesis and
clinical management of gouty arthritis.
J Assoc Physicians India; 2015
2. Kuwabara M. Hyperuricemia,
Cardiovascular Disease, and
Hypertension. Pulse; 2016
3. Hidayat R. Gout dan Hiperurisemia.
Medicinus; 2009
4. Rahmani AH, Aly SM, Ali H, Babiker
AY, Suikar S, Khan AA. Therapeutic
effects of date fruits (Phoenix
dactylifera) in the prevention of
diseases via modulation of anti-
inflammatory, anti-oxidant and anti-
tumour activity. Int J Clin Exp Med;
2014
5. Yasin BR, El-Fawal HAN, Mousa SA.
Date (Phoenix dactylifera)
polyphenolics and other bioactive
compounds: A traditional islamic
remedy’s potential in prevention of cell
damage, cancer therapeutics and
beyond. Int J Mol Sci; 2015
6. Alnajjar, BOM. Computational studies
of natural flavonoids towards the
discovery of a potential xanthine
oxidase inhibitor. Univ Sains Malaysia;
2008
7. Katzung BG. Farmakologi Dasar dan
Klinik. In: Basic and clinical
6
Pharmacology; 2012.
8. Hidayat NT, Perbawani B. Best Method
for the Extraction of Egg Carotenoid
Pigments Golden Egg Snails (Pomacea
canaliculata Lamarck). Int J Chem
Biomol Sci; 2016
9. Ulya S. Pengaruh Pemberian Ekstrak
Daging Buah Kurma Ajwa (Phoenix
Dactylifera L.) Terhadap Kadar
Hemoglobin Pada Mencit (Mus
Musculus) Bunting. Skripsi Oleh :
Syahidatul Ulya Program Studi Biologi;
2018
10. Schunack, W., Mayer, and K., Manfred
H. Senyawa Obat Kimia Farmasi.
Gajah Mada Univ Press Yogyakarta;
1990
11. Mutschler E. Dinamika Obat, Buku
Ajar Farmakologi dan Toksikologi
Edisi Kelima. ITB, Bandung.
1990;217–21.
12. Suhendi A, Nurcahyanti, Muhtadi,
Sutrisna E. Antihyperurisemia activity
of water extract of black seed (Coleus
ambonicus Lour) in balb-c mice and its
standardization. Maj Farm Indones;
2011
13. Zastrow V., Bourne R. Zastrow, V. M.,
Bourne, R. H (2001). Reseptor dan
Farmakodinamika Obat. In: Katzung
BG, editor. Farmakologi Dasar dan
Klinik. 1st ed. Jakarta: Salemba
Medika; 2001. p. 53.