Our Lady of Fatima University

120 McArthur Highway, Marulas, Valenzuela City

College of Medicine
Department of Pharmacology

Protective Effects of Tamarindus indica Ethanolic Extract against

Potassium bromated-induced Hyperuricemia in Male Albino Mice

IN PARTIAL FULFILLMENT OF THE REQUIREMENTS

FOR THE COMPLETION OF THE COURSE
PHARMACOLOGY I

OF THE
COLLEGE OF MEDICINE
OF
OUR LADY OF FATIMA UNIVERSITY

CLUSTERS A & B
April 2018
 CHAPTER I
THE PROBLEM AND ITS BACKGROUND

I. BACKGROUND OF THE STUDY

Gout is a systemic rheumatic condition which manifests as acute and recurring attacks of
severe pain and inflammation affecting peripheral joints (Bardin, 2010). A state of excessive
circulating concentrations of urate, or hyperuricemia, plays a major role in the pathogenesis of this
condition (Schumacher, 2008). As the level of serum uric acid rises and exceeds the physiologic
threshold for uric acid, monosodium urate crystals are precipitated and are deposited around joints.
The deposition of the crystals is responsible for the clinical manifestations that lead to limitations in
the activity of patients suffering from the disease. Numerous risk factors are linked to the development
of gout in men and women have been established. These include genetic factors, dietary factors,
alcohol consumption, metabolic syndrome, hypertension, obesity, diuretic use and renal disease
(Roddy, 2010). According to the Philippine Rheumatology Association (PRA), there are now some 1.6
million Filipinos with gout and the figure is expected to increase in the coming years due to unhealthy
lifestyle of people (Philippine Star, 2015).
Tamarind (Tamarindus indica L.) is a plant native to Africa, but is also found in all tropical
countries including the Philippines. Most of the plant is being used for nutritional and medicinal
purposes, with the pulp being highly valued (Caluwe et al., 2010). The fruit pulp of tamarind has a
sweet acidic taste, mainly due to tartaric acid. Because of this, it is frequently used for seasoning,
sauces, jams, and juices (El-Siddig, 2006).
Aside from its flavor and nutritional value, tamarind has also been used as an herbal medicine
in various countries to treat different diseases. Almost all parts are ideal sources of essential amino
acids and phenolic compounds giving the plant to possess anti-inflammatory, cardio-protective, anti-
diabetic, anti-microbial and anti-oxidant properties, which are just a few of the many medicinal effects
of the T.indica (Kuru, 2014). In various regions in Southeast Asia, the pulp is used as a poultice
applied to the forehead for relief of fever and as a juice for scurvy and cough. In the Philippines, the
leaves are used as an herbal infusion for malarial fever. (Ugoh & Haruna, 2013).
Hence, in line with it being anti-inflammatory, among others, this fruit may be potentially used
in inflammatory disorders, such as gout. Its anti-oxidant property along with its anti-inflammatory
properties may be particularly important in not just counteracting the effects of gout but also
preventing the development of the disease.
II. STATEMENT OF THE PROBLEM
This research aims to determine the protective effects of Tamarindus indica ethanolic extract
against potassium-bromate treated on male albino mice.

Specifically, the study aims to answer the question:

1. Which concentrations of the extract will exhibit a protective effect?
2. Is the tamarind extract comparable to the standard drug, Allopurinol? Is it better than
placebo?

III. HYPOTHESIS

Null Hypothesis (Ho):

The Tamarindicus indica pulp extract will not exhibit protective effects against potassium
bromate-induced hyperuricemia in male albino mice.

Alternative Hypothesis (Ha):

The Tamarindicus indica pulp extract will exhibit protective effects against potassium
bromate-induced hyperuricemia in male albino mice.

IV. SIGNIFICANCE OF THE STUDY

Gout is a common disease that greatly limits the quality of lives patients suffering from it. The
swelling and pain patients experience restricts them from being able to perform even daily activities.
Moreover, this also has long term complications such as nephrolithiasis, erosion of joints, and even
renal failure, to name some. This condition is eventually brought about by an increased production or
decreased excretion of uric acid. It is then important to tackle the root cause of the problem to improve
the lives of people.

Patients with Hyperuricemia

Researches about hyperuricemia may lead to new treatment options that may improve the lives
of patients with the condition. Apart from that, prophylactic measures may also be identified in order
to decrease further incidence of the disease.
Medical Institution
 The study serves as a new related source of information for the continuing experimental
research for hyperuricemia. The research gives information about the protective effects of Tamarindus
indica ethanolic pulp extract against hyperuricemia. Doctors can further continue the study and
provide additional information for the continuity and success of more research studies.
General Public
This research study aims to inform the public regarding the negative effect of hyperuricemia in
the body. This will also inform the public of possible beneficial effects of the sampaloc.
Future Researchers
The research study encourages future researchers to continue and improve the value of this
study.

V. SCOPE AND LIMITATIONS

This study will cover the determination of the protective effect of Tamarindicus indica pulp
ethanolic extract against potassium bromated-induced hyperuricemia in male albino rats. It will also
cover at which concentration will the extract exhibit maximal inhibition. Furthermore, the extract will
also be tested against placebo and Allopurinol. This will not cover the use of other solvents for the
extraction. It will also be limited to the pre-determined concentrations.

VI. DEFINITION OF TERMS

Hyperuricemia - defined operationally as a serum uric acid concentration of >420 umol/L in adult
men and >350 umol/L in adult women

Gout – episodic, severely painful form of inflammatory arthritis due to increased uric acid.
 CHAPTER II
REVIEW OF RELATED LITERATURE

Epidemiology of Gout and Hyperuricemia

Gout is defined as transient attacks of acute arthritis initiated by crystallization of monosodium
urate within and around joints. Almost all types of gout share the common feature of increased levels
of uric acid in the blood, or hyperuricemia (Kumar et al., 2015). The worldwide prevalence of gout
varies around the world. The prevalence of gout is >1% in most developed countries, especially in
Europe and North America. However, it has been noted that prevalence of gout are most common in
Pacific areas, where it was noted to have a higher prevalence in locals living in Pacific countries as
compared to those living in European countries. (Kuo et al., 2015). A data analysis study of the US
National Ambulatory Medical Survey shows that there is an increased likelihood that people of Asian
descent would have higher odds of developing gout as compared to people of white descent (Krishnan
et al., 2014).
In the Philippines, data from the Nutrition and Health Survey yielded a 1.6% occurrence of
gout among adult Filipinos (Li-Yu, 2008). Prevalence studies suggest that gout is more common in
males, and with females affected only after the post-menopausal stage of their lives. (Hernandez, Tee,
and Salido, 2016). In the clinical setting, evidence-based guidelines for management of gout are
available, but it is still far from ideal. A study conducted by Li-Yu and Edar (2017) described various
ways of treatment plans and protocols performed by general practitioners in Metro Manila. It has been
concluded that Gout management has significantly improved in some aspects in accordance to the
2008 Philippine Clinical Practice Guidelines, but still lacks in consistency and heterogeneous patterns
of practice in the community.

Recent Studies in Hyperuricemic Drugs

Plants have increasingly been used for gout and antihyperuricemic studies over the past couple
of years. A study by Wang et al. (2016) showed that the ethanolic and aqueous extract of walnut
(Juglans regia L.) significantly decreased serum uric acid levels in vivo and had high xanthine oxidase
inhibitory action. Sunflower (Helianthus annuus) head extract was found by Li et al. in 2017 to reduce
the inflammatory cells and suppress the swelling of the ankles in rats with acute gout induced by
monosodium urate crystals.
In the Philippines, a recent study by Calderon et al. (2016) showed that an aqueous leaf extract
from papaya (Carica papaya L.) may have a protective influence against hyperuricemia. Three
experimental groups were given the extract with the following dosages: 1, 2, and 3 g/kg BW. Their
results showed that serum uric acid levels were not significantly increased in the experimental groups
compared to the positive control group. In addition to this, renal histopathology performed showed
significant attenuation of acute renal tissue damage in the experimental groups.
The flavonoids found in plants are possible candidates for antihyperuricemic potential. In a
study conducted by Al-Azzawie and Abd (2015), crude flavonoids from ginger (Zingiber officinale)
were given in different doses to male Wistar rats and were found to significantly reduce serum uric
acid levels in hyperuricemic rats in a time-dependent manner. A study by Cheng et al. (2015) on the
antihyperuricemic effect of Lippia nodiflora concluded that flavonoids might be responsible for its uric
acid lowering effect through liver xanthine oxidase/xanthine dehydrogenase inhibition.

Allopurinol
Xanthine Oxidase (XO) inhibitor allopurinol has been the keystone of the clinical management
of gout associated with hyperuricemia for several decades. (Pacher et al., 2006) It is a 1,5-dihydro-4H-
pyrazolo[3,4-d]pyrimidin-4-one which was one of the crown jewels of the respected drug discovery
program at Burroughs Wellcome which also yielded to drugs such as acyclovir, trimethoprim, and the
early antineoplastic compounds thioguanidine and 6-mercaptopurine (6-MP). (Hitching et al., 1963)
XO was recognized to be involved in the formation of uric acid from xanthine and hypoxanthine, and
XO inhibition was thought to inhibit oxidation of 6-MP and potentiate antitumor properties. Thus,
succeeding experiments presented effective regulation of serum and urinary acid. Allopurinol is a
substrate for and competitive inhibitor of the enzyme at lower concentrations and noncompetitive
inhibitor at high concentrations. Oxypurinol on the other hand is responsible for much of its
pharmacological activity as the enzyme’s noncompetitive inhibitor. It is approved by the Food and
Drug Administration in 1966. (Pacher, et al., 2006)

As stated above, the generic treatment for gout as it is the most commonly used management
to lower blood uric acid that has been available for more than 50 years (Walker, 2013). Among heart
failure patients, it is beneficial in reducing uric acid levels, preventing acute gout and act as an
antioxidant (Thanassoulis et. al., 2010). It is also used as medication for patients who have renal uric
acid stones, patients with extreme tophus deposition (> 1,200 mg in 24 hours), patients that are
unresponsive to uricosuric drugs and patients with blood disorders (polycythemia vera, leukemia).
Allopurinol prevents reperfusion injury as well on transplanted organs or organs with blocked blood
perfusion (Craig & Stitzel, 2004). Overall, it showed considerable capacity in the treatment of various
forms of ischemic and other types of tissue and vascular injuries, inflammatory diseases, and chronic
heart failure with its active metabolite oxypurinol, both in experimental animals and in scales of
human clinical trials. (Pacher et al., 2006)

Allopurinol is widely used and generally well-tolerated. However, chronic use of allopurinol is
associated with transient and minor liver test abnormalities and has been linked to acute liver injury
that is typically associated with immunoallergic manifestations. (Chawla et.al, 2005) It is a purine
analogue of xanthine oxidase inhibitor, Xanthine oxidase (XO) is a rate- limiting enzyme that catalyzes
uric acid production and is link to non- alcohol fatty liver disease (NAFLD) and hyperuricemia. XO
inhibitors (allopurinol) significantly decreases uric acid production and attenuates free fatty acid
induced fat accumulation in HepG2 cells causing NAFLD. (Chengfu et. al, 2015).
Moreover, allopurinol, when used in patients with renal insufficiency may have life
threatening toxic effects, such as vasculitis, toxic epidermal necrolysis (TEN), hepatitis, reduced renal
function and bone marrow suppression, known as allopurinol hypersensitivity syndrome (AHS).
Allopurinol is metabolized to oxypurinol, an active metabolite, which is excreted via the kidney into
urine in the same manner as uric acid. Oxypurinol tends to accumulate in patients with renal
prerequisite for the occurrence of AHS. (Horiuchi et. al, 2000)

Experimental Induction of Hyperuricemia using Potassium Bromate

Numerous studies of hyperuricemic induction in an animal experimentation setting are widely
available due to a conso. In a study conducted by Fatimah, C. (2017), potassium bromate has been
linked to increased serum uric acid in albino mice, via gastric gavage. The mechanism of the increase
in serum uric acid levels is believed to be linked to the nature of potassium bromate increasing
oxidative reactions in the test animal, causing the formation of excessive uric acid from the breakdown
of protein compounds. Furthermore, induction of potassium bromate has been linked to an increase in
the activity of xanthine oxidase, the rate-limiting enzyme in the uric acid synthesis pathway. This has
been backed up in a study conducted by Azma et.al (2012), where hyperuricemia on potassium
bromate-induced albino rats have been appeared to be a response to oxidative stress, which may have
reduced catalase activity and increased xanthine oxidase activity, leading to an increase in the serum
uric acid levels of the test animal.
A study by Calderon et al. (2005) was shown to have utilized potassium bromate as the study’s
negative control for the induction of hyperuricemia, and was used as a basis of comparison to
treatment groups via induction at a dose of 200 mg/kg BW dissolved in sterile water per orem by
gastric gavage. The negative control yielded an elevation in the serum uric acid levels of the test
animals, and was compared to the study’s treatment group serum uric acid levels.
Tamarindus indicus and its pharmacologic potentials
Tamarind (Tamarindus indica) from family Fabaceae, sub-family Caesalpiniaceae is a tropical
fruit found in Africa and Asia. In a study of Kumar & Bhattacharya, 2008 it was said that, it is a
multipurpose tree of which almost every part finds at least some use either nutritional or medicinal. Its
various parts such as seeds, root, leaves, bark and fruits have been extremely used in traditional India
and African medication.

Tamarindus indica L. or in Filipino known as Sampalok is a member of Leguminosae family has

always been part of our Filipino cuisine such as sinigang. Tamarind is a woody perennial fruity species
native to Asia and Africa and has been known to be adaptive in different climate, making it a valuable
asset in different food productions, such as jams and beverages (Mohamed, Mohamed, and Ahmed,
2015).

For years, the pulp of the tamarind has been used as an antiscorbutic, laxative, and
carminative. Tamarind has proven to be particularly useful for treating liver and bile disorders. Used in
a gargle for sore throat, as a liniment for rheumatism, applied on inflammations, administered to
alleviate sunstroke and alcoholic intoxication. It’s a part of Vermifuge ointment used to aid the
restoration of sensation in cases of paralysis.

In their admirable effort to consider folk remedies, for so many people who can’t afford
modern prescription drugs, TRAMIL notes that ethanolic extracts are bactericidal, and that the diuretic
pulp inhibits the gram(-) bacteria responsible for urinary infections. This suggests a food pharmacy
approach to cystitis combining the urinary antiseptic properties of cranberry and tamarind in a pleasant
tart beverage. TRAMIL also reports antilipoperoxidative and hepatotropic properties, for the aqueous
leaf extract, approved by TRAMIL for use in jaundice. They note antibacterial, antispasmodic, and
vasodilator properties of the alcoholic extract (DAD, LEG, TRA, WOI).

Khan and Balick (2001) note human studies on tamarind for pain and worms. Tamarind
increased bioavailability of other drugs and decreased pain (JAC7:405). Also, it can kill Bacillus
subtilis, Escherichia coli, and Saccharomyces cerevisiae (De et al., 1999). Sambaiah and Srinivasan
(1989) note that tamarind stimulates N-demethylase activity. It also stimulated liver microsomal
cytochrome p450 dependent aryl hydroxylase.
Chemical compounds with anti-hyperuricemic action
Tamarindus indica is a widely valued fruit with known medicinal uses from leaves to roots but its
fruit pulp is the most used part of the plant. The most outstanding characteristic of tamarind is its sweet
acidic taste, the acid due to mostly tartaric acid (10%) (El-Siddig et al., 1999).
Tartaric is an unusual plant acid formed from the primary carbohydrate products of
photosynthesis, and once formed, it is not metabolically used by the plant (El-Siddig et al., 2006). The
content of tartaric acid does not decrease during fruit ripening, suggesting it is not utilized in fruit
development. At this same time of fruit development; reducing sugars increase to 30-40% giving the
sour fruit a sweeter taste (El-Siddig et al., 2006). As a result, tamarind is known to be simultaneously
the most acidic and sweetest fruit (Lewi & Neelakantan (1964a) and Coronel (1991) both cited in El-
Siddig et al., 2006).
Tamarind pulp typically contains 20.6% water, 3.1% protein, 0.4% fat, 70.8% carbohydrates,
3.0% bre and 2.1% ash (El-Siddig et al., 1999), thus the pulp has a low water content and a high level
of protein, carbohydrates and minerals. Nevertheless, the proximate composition of the tamarind fruit
depends on locality (El-Siddig et al., 2006).
In addition, studies have shown that Tamarindus indica contains various phytochemical contents.
In a study by by Atawodi, Liman, Ottu and Iliemene (2014) the analysis showed that the leaves, fruit
pulp, fruit bark, seeds, roots and stems of the plant are rich in polyphenols and flavonoids. Thus, the
high levels of polyphenols and flavonoids in the fruit pulp of the plant suggest that the therapeutic
efficacy of this pulp in traditional medicine might be through free radical scavenging mechanism.
Furthermore, the ethanolic extract of the tamarind pulp showed presence of more number of
compounds compared to aqueous and methanolic extracts (Y. Jadhav, Dipali & Sahoo, A & Ghosh, Jai
& Ranveer, Dr. Rahul & Mali, Aruna, 2010).
Nwodo, Obiiyeke, Chigor and Okoh (2011) revealed that carbohydrates, reducing sugars, tannins
and saponins were detected in the fruit pulp extract. Flavonoids and Cyanogenic glycosides were also
present in the extract. Anthroquinone was detected in cold water extract of the fruit pulp in addition to
all the ethanolic extracts. Alkaloids were present in the ethanolic extract as well as the cold and hot
water extracts of the fruit pulp. Steroles were not found in the extract and terpenes occurred only in the
ethanolic extract of the fruit pulp.
In another phytochemicals analysis, it contain alkaloids, flavonoids, saponins and tannins,
compounds that have been reported to inhibit bacteria growth and are capable of protecting certain
plants against bacterial infection. Some of these are E. coli, P. aeruginosa, K. pneumoniae and S.
paratyphi A. (Daniyan et al, 2008) It has numerous active constituents, such as phenolic compounds,
cardiac glycosides, L-(-)mallic acid, tartaric acid, the mucilage and pectin, arabinose, xylose,
galactose, glucose, and uronic acid. The ethanolic extract of T. indica exhibited presence of fatty acids
and various essential elements like arsenic, calcium, cadmium, copper, iron, sodium, manganese,
magnesium, potassium, phosphorus, lead, and zinc. Specifically, the pulp contains organic acids, such
as tartaric acid, acetic acid, citric acid, formic acid, malic acid, and succinic acid; amino acids; invert
sugar (25-30%); pectin; protein; fat; some pyrazines (trans-2-hexenal); and some thiazoles (2-
ethylthiazole, 2-methylthiazole) as fragrant; and the seed polysaccharides are found with a main chain
consisting of β-1,4-connected glucose molecules together with xylose (alpha-1,6) and galactose; total
protein; lipids with fatty oils; and some keto acids. In the leaves of the plant, two triterpenes, lupanone
and lupeol were found. (Bhadoriya, 2011)

OTHER PHARMACOLOGICAL ACTIVITY

ANTIOXIDANT
A study by Sudjaroen et al., shown that the seed and pericarp of Tamarindus indica contain
phenolic antioxidant compound Four anti-oxidative compounds were isolated and identified from the
seed coats: phenolic antioxidants, such as 2-hydroxy-3’, 4’-dihydroxyacetophenone, methyl 3,4-
dihydroxybenzoate, 3,4-dihydroxyphenyl acetate and epicatechin (Tsuda et al., 2004). These
antioxidants may be used for increasing shelf life of food products and improving the stability of lipids
and lipid-containing foods by preventing loss of sensory and nutritional quality by preventing lipid
peroxidation.

ANTI-INFLAMMATORY
A serine proteinase inhibitor, providing high activity against HNE was detected, isolated and
purified from tamarind seeds (Fooket al., 2005). According to Shaikh et al., World J Pharm Sci 2017;
5(2): 121-133,result showed that the plant exhibit anti-inflammatory property from aqueous ethanol
and chloroform extracts of T. indica thsat were evaluated in mice (ear edema induced by arachidonic
acid) and rats(subplantar oedema induced by carrageenan) after topical or i.p.
administration,respectively.

ANTI-DIABETIC ACTIVITY
Frequent research on aqueous extracts of seeds has shown a strong anti-diabetic effect in rats
(Maitin et al. (2004) cited in El-Siddig et al., 2006). Similarly, hyperlipidemia was found to be
reduced, measured by different contents of cholesterol. This rat model may shed some light on the
basis of ancient herbal
therapy in India.

ANTIMICROBIAL ACTIVITY
According to Al-Fatimi and collaborators (2007), in an agar diffusion assay, extracts from T.
indica flowers showed antibacterial activity against four bacteria tested (Staphylococcus aureus,
Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa). Vaghasiya et al., studied that the
Methanol and acetone extracts have shown significant antimicrobial activity against Klebsiella
pneumonia by agar disk diffusion method. Extracts from the fruit appear promising as a potential
fungicidal agent against cultures of Aspergillus niger and Candida albicans (El-Siddig et al., 1999; El-
Siddig et al., 2006). The extract also exhibited anti-parasitic activity against Bursaphelenchus
xylphilusm and Pheretima posthuma (Mackeen et al. (1997) cited in El-Siddig et al., 1999; Mute et.
al.).

Preparation of Tamarindus indicus Extract

Proper preparation of T. indicus extract is important in doing this study. From storage up to the
extraction of significant compounds from it is strictly monitored. According to Obelesu, M., et.al
(2011), ripening of T. indicus gives a significant increase in reducing sugars and lysine which hinder
the enzymatic browning known as Maillard Reaction. This reaction can lead to the formation of
melanoidins and destruction of amino acids.
For the extraction of active compounds of T. indicus pulp, the use of aqueous ethanol (50%) was
found to be the best solvent.
 CHAPTER III

Research Methods

This chapter presents the materials and test procedures that will be used in the study of anti-
hyperuricemic potential of T.indica L. fruit pulp in Potassium Bromate induced Albino mice.

A. Plant Extract
1. Collection and Authentication
T.indica L. fruit pulp were collected from Balintawak Market, Quezon City. The plant
sample was submitted to Bureau of Plant Industry for Identification and Authentication.
2. Extraction
The fruit was washed to remove dirt and impurities. The peel or skin of the fruit was
removed and the pulp was removed and seeds were discarded. The fruit pulp was dried in an oven for
1 day under 40 degrees Celsius and was subjected to extraction with 50% ethanol with a ratio of 1:3. A
kilogram of fruit pulp was extracted with 3 liters of 50% ethanol by maceration guided by magnetic
stirring for 12 hours. After collecting the extract, the solution underwent filtration and excess solvent
was removed under reduced pressure in a rotary evaporator. The final extract was collected and
preserved to prevent contamination and early degradation until used for further studies.

• The tamarind was purchased and fruit pulp was collected

• The fruit pulp was obtained and oven dried for 48 hours under 40 degrees celcius

• After drying, Fruit pulp was subjected to extraction by maceration using 50%
ethanol with a ratio of 1:3

• After extraction, the solution was filtrated

• The excess solvent was removed using a Rotary evaporator under reduced
pressure after collecting the extract

• Final extract was stored in a sealed glass container

Figure 1. Schematic Diagram of Methodology

B. Materials
1. Test Animals
Healthy Male Albino mice, weighing in between 150- 220 grams, were purchased from
the Food and Drug Administration – Muntinlupa City. The animals were housed in an
accredited animal handling facility in Marulas, Valenzuela. The rats were acclimatized for
seven days at a temperature of 18 degrees celcius to 26 degrees celcius, with 12 hours of
light and dark cycle. The animals were fed a standard pellet diet and have free access to
water.
Thirty albino mice were selected and divided further to five groups:.
Table 1. Six groups of test animals
Group Day 1 - 14 Day 14
Group 1 Negative Control Water None
Group 2 Placebo Group Water KBrO3
Group 3 Positive Control Allopurinol KBrO3
Group 4 (200 mg/kg BW) Tamarind extract KBrO3
Group 5 (300 mg/kg BW) Tamarind extract KBrO3
Group 6 (400 mg/kg BW) Tamarind extract KBrO3

Table 1 shows the treatment for the different groups. Group 1 (Negative Control) was only
given water for the first 14 days and on the 14th day, nothing will be given. Group 2
(Placebo) was only given water for first fourteen days and KBrO3 on the 14th day. Group 3
(Positive Control) was given Allopurinol for the first 14 days and on the 14th day KBrO3.
All the next succeeding groups will be given the tamarind extract on the first fourteen days
with varying doses from low, medium and high dose and on the 14th day the KBrO3 will
also be given

C. Evaluation of anti – hyper uricemic potential of plant extract

1. Induction of Hyperuricemia using Potassium Bromate
Seventy-five minutes after the final administration of plant extract, potassium bromate was
given at a dose of 200mg/kg BW, dissolved in distilled water per orem by gastric lavage.
2. Blood Uric Acid Determination
Blood was collected from the tail of the rat on Day 0, intraocularly on Day 7 and from the
Jugular vein at day 14 post induction of KBrO3. Blood uric acid levels were determined
immediately before and three hours post induction of KBrO3.The samples were brought to Hi
– Precision Laboratories.
 D. Statistical Analysis
Paired t-test was used to detect if there was a significant difference in the blood uric acid levels per
point in time within every group. In order to determine different treatment groups, uric acid
measurements were analyzed using two-way ANOVA. If there is a significant difference, this will be
followed up by Tukey’s test.
 REFERENCES
1. Al-Azzawie, H. F., & Abd, S. A. (2015). Effects of Crude Flavonoids from Ginger (Zingiber
officinale), on Serum Uric Acid Levels, Biomarkers of Oxidative Stress and Xanthine Oxidase
Activity in Oxonate-Induced Hyperuricemic Rats. International Journal of Advanced
Research,3(10), 1033-1039.
2. Atawodi, S. E., Liman, M. L., Ottu, J. O. and Iliemene, U.D. (2014). Total Polyphenols,
Flavonoids and Antioxidant Properties of Different Parts of Tamarindus indica Linn of
Nigerian Origin. Annual Research & Review in Biology 4(24): 4273-4283, 2014
3. Calderon, P. E., Juan, C. S., Pedro, M. G. S., Reyes, A. M., Salom, P. J., Sanchez, A. R., ... &
Santos, D. (2016, June). Protective influence of Carica papaya L. aqueous leaf extract against
hyperuricemia and acute renal injury in a murine model. In AIP Conference Proceedings (Vol.
1744, No. 1, p. 020043). AIP Publishing.
4. Caluwé, E. D., Halamová, K., & Damme, P. V. (2010). Tamarindus indica L. – A review of
traditional uses, phytochemistry and pharmacology. Afrika Focus, 23(1), 53-83.
5. Cheng, L., Murugaiyah, V., & Chan, K. (2015). Flavonoids and phenylethanoid glycosides
from Lippia nodiflora as promising antihyperuricemic agents and elucidation of their
mechanism of action. Journal of Ethnopharmacology,176, 485-493.
doi:10.1016/j.jep.2015.11.025
6. Edar, M. and Li-Yu (2017), J.: Clinical Practice in Gout Management Among Filipino General
Care Practitioners, JMUST Journal of Medicine UST 10.1000/2546-1621.017.0052
7. El-Siddig, K., Ebert, G., Lüdders, P. (1999). Tamarind (Tamarindus indica L.): a
Review on a Multipurpose Tree with Promising Future in the Sudan. Journal of
Applied Botany – Angewandte Botanik, 73, 202-205.
8. El-Siddig, K. (2006). Tamarind: Tamarindus indica L. Southampton: Internat. Centre for
Underutilised Crops, Univ. of Southampton, p. 24 – 26.
9. Fatimah, Cut(2017): The Effectiveness of Bandotan Herb (Ageratum Conyzoides L.) as Blood
Uric Acid Levels Reduction in Mencit with Allopurinol Comparison, International Journal of
Phytopharmacy May-Jun 2017; Vol. 7 (3): pp. 23-29. 23 IJPP | Volume 7 | Issue 3 | 2017
https://doi.org/10.7439/ijpp
10. Garcia, L. L., et al. (2003). Philippine vegetable drugs in common use: their chemical
constants part II. Philippine Journal of Science, 132 (2), 103-108.
11. Hernandez, A.T.,Tee, K. and Salido, E (2016).: Clinical Profile of Filipino Female Patients
with Gout Seen in Adult Rheumatology Clinics in the National Capital Region, Philippines,
Philippine Journal of Internal Medicine, Volume 54 Number 3 July-Sept., 2016
12. Kumar, et. al: Pathologic Basis of Disease, pg. 1214-1217, 9th Edition, 2015, ISBN:
9781455726134, Elsevier
13. Kuo CF, Grainge MJ, Zhang W, Doherty M (2015): Global epidemiology of gout: prevalence,
incidence and risk factors. Nat Rev Rheumatol. 2015 Nov;11(11):649-62. doi:
10.1038/nrrheum.2015.91. Epub 2015 Jul 7.
14. Li, L., Teng, M., Liu, Y., Qu, Y., Zhang, Y., Lin, F., & Wang, D. (2017). Anti-Gouty Arthritis
and Antihyperuricemia Effects of Sunflower (Helianthus annuus) Head Extract in Gouty and
Hyperuricemia Animal Models. BioMed Research International,2017, 1-9.
doi:10.1155/2017/5852076
15. Li-Yu, JT (2008): Relationship of Gout and Hyperuricemia in Cardiovascular and Renal
Morbidity: 2003 National Nutrition and Health Survey (NNHeS). Phil Journal of Internal
Medicine, 46: 279-285, Nov-Dec, 2008
16. Mohamed HA, Mohamed BE, Ahmed KE.(2015).Physicochemical Properties of Tamarind
(Tamarindus indica) Seed Polysaccharides. J Food Process Technol 6:452. doi:10.4172/2157-
7110.1000452
17. Muzaffar, K., & Kumar, P. (2018). Effect of Process Parameters on Extraction of Pulp from
Tamarind Fruit. Department of Food Engineering & Technology, Sant Longowal, Sangrur,
Punjab, India. Conceptual Frame Work & Innovations in Agroecology and Food Sciences
ISBN: 978-81-930585-1-0
18. Prasad P, Krishnan E. Filipino Gout(2014): A Review. Arthritis Care and Research.
2014;66(3):337-343. doi:10.1002/acr.22118.
19. Wang, X., Zhao, M., Su, G., Cai, M., Sun-Waterhouse, D., Zhou, C., & Lin, L. (2015).
Antihyperuricemic activities of an ethanolic and aqueous extract of Walnut (Juglans regia L.)
shell and a new aldehyde xanthine oxidase inhibitor. International Journal of Food Science &
Technology,51(2), 453-460. doi:10.1111/ijfs.12995
20. Ugoh, S. C., Haruna, I. M. (2013). Phytochemical screening and antibacterial activity of the
fruit and leaf extracts of Tamarindus indica (Linn.). Report and Opinion 2013 5(8), 20-27.
21. Chong,U.R., et al. (2013). Effects of Tamarindus indica Fruit Pulp Extract on Abundance of
HepG2 Cell Lysate Proteins and Their Possible Consequential Impact on Metabolism and
Inflammation. BioMed Research International, Article ID 459017
22. Bandawane, D., et al. (2013). Evaluation of anti-inflammatory and analgesic activity of
tamarind (Tamarindus indica L.) seeds. International Journal of Pharmacy and Pharmaceutical
Sciences 5(4), 623-629
23. Kuru, Pinar., (2014). Tamarindus indica and its health related effects. Asian Pacific Journal of
Tropical Biomedicine 2014 4(9), 676-681
24. Sole, S.S., Srinivasan, B.P., & Akarte, A.S. (2013). Anti-inflammatory action of Tamarind
seeds reduces hyperglycemic excursion by repressing pancreatic β-cell damage and
normalizing SREBP-1c concentration. New Delhi, India.
25. Sundaram, M.S., et.al., (2015). Tamarind Seed (Tamarindus indica) Extract Ameliorates
Adjuvant-Induced Arthritis via Regulating the Mediators of Cartilage/Bone Degeneration,
Inflammation and Oxidative Stress. Mysore, India.
26. Richette P, Bardin T. Gout. Lancet 2010;375:318–28. Schumacher HR Jr. The pathogenesis of
gout. Cleve Clin J Med 2008;75(Suppl 5):S2–4.
27. Edward Roddy and Michael Doherty, Arthritis Res Ther. 2010; 12(6): 223. Published online
2010 Dec 21. doi: 10.1186/ar3199
28. Y. Jadhav, Dipali & Sahoo, A & Ghosh, Jai & Ranveer, Dr. Rahul & Mali, Aruna.
(2010). Phytochemical Detection and in vitro Evaluation of Tamarind Fruit Pulp
for Potential Antimicrobial Activity. International Journal of Tropical Medicine. 5. 68-72.
10.3923/ijtmed.2010.68.72.
29. Nwodo , U.N., Obiiyeke , G.E., Chigor, V.N. , and Okoh A.I.. (2011). Assessment of
Tamarindus indica Extracts for Antibacterial Activity. Int. J. Mol. Sci. 2011, 12, 6385-
6396; doi:10.3390/ijms12106385
30. Pacher P., Nivorozhkin A., Szabó C. (2006). Therapeutic Effects of Xanthine Oxidase
Inhibitors: Renaissance Half a Century after the Discovery of Allopurinol. Pharmacological
Reviews. 58(1):87-114
31. Thannassoulis, G., Brophy, J., Richard, H. and Pilote, L. (2010) Gout Allopurinol Use, and
Heart Failure Outcomes. Arch Intern Med. 170(15):1358-1364
32. Walker, T. (2013) Allopurinol Alone Found Inadequate for Gout Control. UBM Medica
33. Craig C.R. & Stitzel, R.E. (2004). Modern Pharmacology with Clinical Applications (6th ed).
USA: Maryland.