Thesis 2 (Word)

ARELLANO UNIVERSITY - COLLEGE OF PHARMACY PAGE 1
GLUCOSE-LOWERING PROPERTY OF Citrofortunella microcarpa
FRUIT PEEL USING ALLOXAN-INDUCED HYPERGYCEMIC RATS
Cuntapay, Geneece O.
Ms. Regine Gonzales, RPh
Arellano University – College of Pharmacy
ABSTRACT
The study was to determine the glucose-lowering property of petroleum ether extract
from the peel of Citrofortunella microcarpa fruit peel in alloxan induced hyperglycemic
rats. First, the researcher used Alloxan monohydrate to intraperitonially induce
hyperglycemia on twenty sprague-dawley rats. Then, the rats were randomly grouped
into 4. For Group A, the researcher used Normal saline solution (NSS) as the negative
control; For Groups B and C, the researcher used Citrofortunella microcarpa fruit peel
extract (250mg/kg for Group B and 500 mg/kg for group C) and is orally
administered; Lastly, for Group D, the researcher used Metformin, a common anti-
diabetic agent as the positive control. During the research, it was found out that
locally grown Citrofortunella microcarpa is slightly miscible with ethanol and distilled
water, and completely miscible with petroleum ether, chloroform and acetone. As for
the Organoleptic test, the mixture resulted into dark green color, sour taste, semi-solid
appearance and pungent odor; and the percentage yield is 2.52%. Based on the
pytochemical testing extract of Citrofortunella microcarpa contains flavonoids.And as
for the solubility testing Citrofortunella microcarpa is soluble in ethyl alcohol,petroleum
ether, chloroform, and acetone while insoluble in distilled water. The petroleum ether
extract of Citrofortunella microcarpa fruit peel has a glucose lowering property at
(p<0.05). It was found out in the research that the glucose lowering property of
Citrofortunella microcarpawas more effective at 250mg/kg as compared to 500mg/kg.
Keywords: glucose-lowering, alloxan,Citrofortunella microcarpa, hyperglycemia,

diabetes
INTRODUCTION
Diabetes mellitus is a chronic disease which is caused by either inherited illness or
acquired deficiency in production of the hormone insulin and its subsequent inability
to regulate blood glucose level. (Sriparna, et. al., 2013) The chronic hyperglycemia of
diabetes is associated with long-term damage, dysfunction and failure to various
organs, especially the eyes, nerves, heart, and blood vessels. (Kalaivanan, K. et. al.
2011). In modern medicine, there are many treatment options that attempt to cure
diabetes like insulin and insulin analogs, alpha-glucosidase inhibitors, sulfonylureas,
biguanides and amylin analogs, however, aside from the fact that they are not cost-
friendly, certain adverse effects like hypoglycemia, liver damage, lactic acidosis and
diarrhea were also observed. Therefore, it is a necessity to look for newer agents that
are economically viable and locally available to lower blood glucose like Citrofortunella
microcarpa.
Nature has been a good source of medicinal substances for thousands of years, and
natural products continue to play a vital role in the primary care of almost 80% of the
world’s population especially in underdeveloped or developing country like the
Philippines. Citrofortunella microcarpa also known as calamondin comes from the
family Rutaceae. The Citrofortunella microcarpa bears a small citrus fruit that is used
to flavor foods and drinks. Citrofortunella microcarpa, is a smooth and slightly spiny
plant, growing to a height of 3 to 5 meters. Its leaflets are eliptic to oblongeliptic, 4 to 8
centimeters long. Its petioles are very narrowly or scarcely winged and about
1centimeter long. Its flowers are axillary, solitary, rarely in pairs, white, and short-
stalked. Its fruits are yellow whenripe, nearly spherical, 2 to 3.5 centimeters diameter,
6 to 7 celled, and thin skinned. The fruit’s skin or peel is yellowish green or yellow,
loosely adhering to the fresh. The flesh contains a few light orange seeds. Globally,
Citrus fruits have the highest production within the fruit category and are
commercially grown in more than 50 countries (Ladaniya, 2008). It is also
commercially available in any countries. It is accessible and inexpensive.
Citrofortunella microcarpa is said to be a wonder fruit that help lower blood glucose
level in our body and is traditionally believed to be a remedy for high blood glucose
level. Metformin, a commonly prescribed oral hypoglycemic agent (OHA) is widely used
in the management of type 2 diabetes mellitus. It can lower blood glucose
concentration without causing hypoglycemia. (Lee, M. G., et. al., 2008)
The health-promoting properties of citrus fruits have been ascribed to their inherent
phenolic compounds including coumarins, flavanoids, lignins, phenolic acids and
tannins (Xu, Ye, Liu, Ma, & Chen, 2008). Several studies on phenolic acids (e.g.,
caffeic, p-coumaric, ferulic and sinapic acids) in citrus fruits have been confined to the
understanding of their distributions among the cultivars during maturation and
exploration of their nutritional properties (Kelebek, 2010; Kelebek, Canbas, &Selli,
2008; Peleg, Naim, Rouseff, & Zehavi1991; Rapisarda, Carollo, Fallico, Tomaselli,
&Maccarone, 1998). To
date, there is noreport of phenolic acids content in locally available Citrofortunella

microcarpa peel.
According to previous studies, the peels of citrus fruits are rich in pectin, which are
known to possess blood sugar lowering, and cholesterol lowering properties. On basis
of the preliminary phytochemical tests conducted on fresh fruit peels of Citrofortunella
limetta, the methanol extract was found to be rich in flavonoids (well-known
antioxidants) and therefore was selected for the hypoglycemic activity (Sriparna,
KunduSen et al., 2011)
OBJECTIVE
Specifically, the main objective of this study is to determine the glucose lowering
property of Citrofortunella microcarpa L. fruit peel in alloxan induced diabetic rats:
Determine the percentage yield that will obtained from the sample that will be used in
the research study
Determine the solubility of Citrofortunella microcarpa peel extract.
Determine the concentration of Citrofortunella microcarpa peel extract exhibited

hypoglycemic effect
Determine if there is no significant difference on the hypoglycemic effect of
Citrofortunella microcarpa with standard drug Metfromin HCl.
STATEMENT OF THE PROBLEM
Moreover, to establish concrete evidence for such claims, the study ought to answer
the following questions:
What is the percentage yield of Citrofortunella microcarpa?
Is flavonoid present in Citrofortunella microcarpa peel extract?
What concentration of Citrofortunella microcarpa peel extract exhibits a hypoglycemic

effect?
Is there a significant difference between the hypoglycemic activity of Citrofortunella

microcarpa and the standard drug Metformin HCl?
REVIEW OF THE RELATED LITERATURE
This chapter discusses about the different related literature and studies which are
related to the present study that serves as a guide for the research questions.
Taxonomical Classification
The botanical classification of Anamirta cocculus is classified under family

Menispermaceae of the order Ranunculales, (Figure 2.1) Kingdom Plantae – Plants
Subkingdom Tracheobionta – Vascular plants Superdivision Spermatophyta – Seed plants
Division Magnoliophyta – Flowering plants
Class Magnoliopsida – Dicotyledons
Subclass Rosidae Order
Sapindales
Family Rutaceae– Rue family

Botanical Description
Citrofortunella microcarpa is a smooth and slightly spiny plant, growing to a height of 3

to 5 meters. Leaflets are eliptic to oblong-eliptic, 4 to 8 centimeters long, Its petioles
are very narrowly or scarcely winged about 1 centimeter long, while is flowers are
axillary, solitary, rarely in pairs, white,
and short-stalked. Its fruit is yellow when ripe, nearly spherical measuring 2 to 3.5
centimeters in diameter, 6 to 7 celled, and thin skinned. The skin or peel is yellowish
green or yellow, loosely adhering to the fresh. The flesh contains a few light orange
seeds. Citrus fruits globally have the highest production within the fruit category and
are commercially grown in more than 50 countries (Ladaniya, 2008)
Figure 1. Citrofortunella microcarpa (Calamansi)

Folkloric Uses
Citrofortunella microcarpa has many traditional and medicinal uses and nowadays
people are still adapting it, the most common use of the fruit’s extract is stain
remover, an aromatic bath, juice mixed with gogo, warm kalamansi-ade drunk for
cough, cold and sore throat, for nausea and fainting, rind is squeezed near nostril to
inhale it also applied externally for itching.
In Higanoon, a tribe of Mindanao use decoction of leaves to lower hypertension. Juice

from partly roasted fruits used for coughs and colds. Fruits crushed with bark of
Entadaphaseoloides used as hair shampoo, for itching and to stimulate hair growth.
Juice of fruit used for Acne vulgaris and Pruritis vulvae. In Malaysia, it is used as an
antidote for poison. Poultice of pandanus leaves, mixed with salt and juice of citrus
microcarpa, for abscesses.
In Malaya, combined with pepper to help expel phlegm. Root
used at childbirth. Leaf oil used as carminative, with an effect stronger than
peppermint oil. Fruit crushed with bark of Entadaphaseoloides used as hair shampoo,
for itching and to stimulate hair growth.
Chemical Constituents
A study on the constituents of Citrofortunella microcarpa or whole fruit essential oil

yielded 54 compounds, including flavonoids ,13 monoterpenes, 7 monoterpene
alcohols, 1 monoterpene oxide, 4 monoterpene aldehydes, 2 monoterpene ketones, 4
monoterpene esters, 12 sesquiterpenes, 3 alipathic alcohols, 6 aliphatic aldehydes,
and 2 alipathic esters, with limonene and Bmyrcene as major compounds. Leaves:
volatile oil; 0.90-1.06% rind: Aldehydes sesquiterpenes B-pinene linalooi linely1
acetate tannin glycosides and cyanogenetic substances.
Pharmacological Uses
Citrofortunella microcarpa has been demonstrated to exhibit antioxidant function and

tyrosinase inhibitory activity, which might be attributed to its flavonoid compounds.
To improve their application, the flavonoid compositions and antioxidant activity of
calamansi extract, prepared different solvent were investigated. The results showed
that total phenolic and flavonoid contents extract from peel of calamansi were higher
than from pulp. Studies have also shown that it has antimicrobial activity (Philippine
Dermatological Society, et al., 2008).
It also has anti- inflammatory activity to relieve pain in arthritis and hemorrhoids. A
heavy metal poisoning chelator. It also lowers cholesterol. These pharmacologic
activities are attributed to its pectin. Antianxiety or Antidepressant; Study provides
evidence that the smelling of essential oils of Citrofortunella hystrix and Citrofortunella
microcarpa confer anxiolytic effect. It concludes that essential oils of the Citrus family
may affect behavior. (Che Rugayah et., al)
Hepatoprotective, peel extract exhibited hepatoprotective activity against

Acetaminophen-induced liver disease in male Sprague Dawley rats, comparable to
commercially available silymarin preparation (International Journal of Chemical and
Environmental Engineering, December 2010). Found that rind of calamansi plant
contains tannins, which ca be extracted by
maceration with water. Anti-inflammatory property is effective in increasing dose of

250, 500 and 1000 mg/kg. (Donya Alinejhad et. al. 2015) Citrofortunella microcarpa
peel extract exhibits both antiangiogenic and antioxidant properties. They recommend
development of anti-cancer therapy envisaged, antiangiogenic and antioxidant
bioassays of the extract using tumor-positive animal. (Mary Jane G. Barluado et. al.
(2013).
Related Literature
Diabetes a condition where the pancreas produce insufficient amount of insulin or

when the body unable efficiently use the insulin or both (WHO et al., 2012)
Types of Diabetes
Type I Diabetes
It occurs when the pancreas cannot make insulin. Everyone with type 1 diabetes
requires insulin injections.
Type II Diabetes
It occurs when the pancreas does not make enough insulin or the body does not use
insulin properly. It usually occurs in adults, although in some cases children may be
affected. People with type 2 diabetes usually have a family history of this condition and
are most often overweight. People with type 2 diabetes may eventually need insulin
injections. This condition occurs most
commonly in people of First Nations descent, Hispanics, and North Americans of

African descent.
Causes of Type I Diabetes
Animal models for type 1 diabetes range from animals with spontaneously developing
autoimmune diabetes to chemical ablation of the pancreatic beta cells.
Causes of Prediabetes and Type II Diabetes
Type 2 diabetes is characterized by insulin resistance and the inability of the beta cell
to sufficiently compensate. Therefore, animal models of type 2 diabetes tend to include
models of insulin resistance and/or models of beta cell failure. In this study, albino
rats type 2 diabetes are obese, reflecting the human condition where obesity is closely
linked to type 2 diabetes development. (Yoshida et al., 2010; Gault et al., 2011; Park et
al., 2011).
Sign and Symptoms
There are many ways of determining that you have a Diabetes, here are the signs and
symptoms of the disease, first is increased thirst, increased hunger (especially after
eating) frequent urination or urine infections, unexplained weight loss, fatigue (weak,
tired feeling) blurred vision, headaches, loss of consciousness.
Diagnosis and Treatment
In fasting, blood glucose is between 6.1 mmol/L and 6.9 mmol/L it may have a
condition known as impaired fasting glucose or prediabetes, If the level of glucose in
blood after 8 hours of fasting is 7.0 mmol/L or higher this can be diagnose as diabetes
which may later develop into diabetes. It can also be diagnosed with a random blood
glucose level. This is a blood glucose level taken any time of the day without regard to
meals.
Alloxan monohydrate – Inducer
The mechanism of alloxan action has been thoroughly studied which currently can be
characterized quite well. Several experimental studies have demonstrated that alloxan
evokes a sudden rise in insulin secretion in the presence or absence of glucose which
appeared just after alloxan treatment. 32-33 This particular alloxan-induced insulin
release occurs for short duration followed by the complete suppression of the islet
response to glucose even when high concentrations of glucose were used.21 Further,
the alloxan action in the pancreas is preceded by its rapid uptake by pancreatic beta
cells that have been proposed to be one of the important features determining alloxan
diabetogenicity. Moreover, in pancreatic beta cells, the reduction process occurs in the
presence of different reducing agents like reduced glutathione (GSH), cysteine,
ascorbate and protein-bound sulfhydryl (-SH) groups. 34-35 Alloxan reacts with two
-SH groups in the sugar binding site of glucokinase resulting in the formation of the
disulfide bond and inactivation of the enzyme. As a result of alloxan reduction, dialuric
acid is formed which is then re-
oxidized back to alloxanestablishing a redox cycle for the generation of reactive oxygen
species (ROS) and superoxide radicals. The superoxide radicals liberate ferric ions from
ferritin and reduce them to ferrous and ferric ions. In addition, superoxide radicals
undergo dismutation to yield hydrogen peroxide (H2O2) in the presence of superoxide
dismutase. As a result, highly reactive hydroxyl radicals are formed according to the
Fenton reaction in the presence of ferrous and H2O2. Another mechanism that has
been reported is the effect of ROS on the DNA of pancreatic islets. The fragmentation of
DNA takes place in the beta cells exposed to alloxan that causes DNA damage, which
stimulates poly ADP ribosylation, a process participating in DNA repair. In addition,
the disturbance in intracellular calcium homeostasis has also been reported to
constitute an important step in the diabetogenic action of alloxan. It has been noted
that alloxan elevates cytosolic free Ca2+ concentration in the beta cells of pancreatic
islets.40 The calcium influx is resulted from the ability of alloxan to depolarize
pancreatic beta cells that further opens voltage dependent calcium channels and
enhances calcium entry into pancreatic cells. The increased concentration of Ca2+ ion
further contributes to supraphysiological insulin release that along with ROS has been
noted to ultimately cause damage of beta cells of pancreatic islets.
( Depomed Inc. et,. Al 2016)

Figure 2. Alloxan Monohydrate Chemical Structure, (Pubchem, 2005)
Metformin Hydrochloride - Standard Drug
Metformin is an antihyperglycemic agent, which improves glucose tolerance in patients

with type 2 diabetes, lowering both basal and postprandial plasma glucose. Its
pharmacologic mechanisms of action are different from other classes of oral
antihyperglycemic agents. Metformin decreases hepatic glucose production, decreases
intestinal absorption of glucose, and improves insulin sensitivity by increasing
peripheral glucose uptake and utilization. Unlike sulfonylureas, metformin does not
produce hypoglycemia in either patients with type 2 diabetes or normal subjects and
does not cause hyperinsulinemia. With metformin therapy, insulin secretion remains
unchanged while fasting insulin levels and daylong plasma insulin response may
actually decrease. ( Depomed Inc. et,. Al 2016)
Figure 3. Metformin Chemical Structure, (Pubchem, 2005)

Related Studies
Citrofortunella microcarpa or better known as Calamansi/Kalamansi is a commercially

available citrus fruit in the Philippines. Although it is easily accessible and
inexpensive, studies on its ability to lower blood glucose levels in the body are
insufficient. Blood glucose levels are vital in the prevalence of certain diseases due to
its role in the mechanisms of the body. The risk of diabetes mellitus (DM) is heightened
when there is hyperglycemia or uncontrollable increase of blood glucose level in the
body and the insulin in the body is unable to balance out the sugar levels. Higher
glucose levels also increase the risks of liver disease, obesity and dementia. (Crane et
al., 2013).
Citrofortunella microcarpa also contains lignin, a fiber-like component. This tends to

promote satiety and may reduce the rate of glucose uptake followingconsumption of
glycemic (available) carbohydrate, thus helping to prevent a surge in blood glucose
levels. (Whitney &Rolfes, 1999).
Citrus extracts not only slow glucose uptake, but also inhibit the movement or
transport of glucose through the intestines and liver (Day, 2015). Citrus also contains
lignin, a fiber-like component. Citrus extracts not only slow glucose uptake, but also
inhibit the movement or transport of glucose through the intestines and liver (Day,
2015).
Citrus fruits also contain Vitamin C which is an important for boosting the immune
system against other diseases. Vitamin C helps improve the absorption of calcium and
iron by the body which is helpful for diabetics because of their bodies inability to
absorb nutrients. An article from Philippine Star 2014 stated that citrus fruits alleviate
inflammatory conditions like asthma, osteoarthritis and rheumatoid arthritis. It also
protects the heart and boosts the immune system. That is why it is good in preventing
diseases.
Quisumbing recorded the following constituents: Leaves: volatile oil; 0.90-1.06% rind:
Aldehydes sesquiterpense â-pinene linalooi linely acetate tannin glycosides and
cyanogenetic substances. He reported that the juice of the fruit is used in removing ink
stains from garments and for washing the hair of ladies and can be utilized for
blanching spots and as an accessible medication for itching. (Sriparna, KunduSen et
al., 2011)
Citrtrofortunella microcarpa or Citrus mitis, also known as 'limaukasturi' is an edible

plant and have potential as antibacterial agent. The presence of bioactive compound in
limaukasturi peels extract have possessed antibacterial property by killing or inhibits
the growth of pathogenic bacterial. In this research, limaukasturi peels were dried and
extracted using soxhlet extraction method. The solvents used for the extraction were
methanol, ethanol, acetone, hexane and water. The antibacterial effect of limaukasturi
peels extract was evaluated on Gram-positive bacteria like Bacillus subtilis
and Streptococcus spp., and Gram-negative bacteria such as Escherichia coli and
Salmonella spp. The extracted products were subjected to antibacterial susceptibility
assay by agar well-diffusion method with measurement diameter of inhibition zone
around the extracts. From the result, methanol extracts showed the highest inhibition
zone compared to other solvent extracts. The zone of inhibition of methanol extracts on
E. coli, Salmonella spp., B. subtilis, and Streptococcus spp. were 19 mm, 17 mm, 13
mm and 12 mm, respectively. Overall, the bacterial are susceptible and moderately
susceptible to methanol extracts whereas the other extracts of ethanol, acetone,
hexane and water showed low inhibition and no inhibition effects towards four
bacterial tested. Thus, the results obtained in the present study suggest that methanol
extracts of limaukasturi peels can be used for further study in treating diseases caused
by the test organisms. Keywords: Limaukasturi, peels, antibacterial properties, soxhlet
extraction, agar-well diffusion method. (Rubiatul, A.S., et al. )
Collection of Citrofortunella microcarpa
Authentication, Pre paration and Drying
of the f ruit peel
Extraction
Petroleum Ether Extractives

Physical Te sts
Presence of Fl avonoids

Organo leptic
Alkaline r eagent
evaluat ion
test
Solubili ty
Lead acet ate test


Ferric chl oride test

Biological Testing
Acclimatization of experimental animals
Induction of Hype rglycemia
Testing of Pharma cological effect

Statistical Treatment
and Interpretation
Figure 4. Paradigm of the Study

METHODOLOGY
To determine if Citrofortunella microcarpa, L. fruit peel has a glucose lowering property,

the researcher used this experimental design of research. The researcher, used alloxan
monohydrate induced diabetic rats. The test animals of the study were subjected to
some type of treatment and condition. The control and treatment groups differ in terms
of the experimental treatment they received.
Collection and Preparation of Plant Material
Citrofortunella microcarpa fruits were obtained from the province of Nueva Vizcaya in
the months of July-September 2017. Fresh fruit peels of Citrofortunella microcarpa
were washed and dried for 2-3 days. 500g of plant material were cut into small pieces
and macerated for 3 days in petroleum ether to further extract the desired
phytochemical constituents. The mixtures were filtered using Whatman filter paper.
The filtrate were concentrated using rotary evaporator at 45 degrees Celsius before
water bath to remove the remaining solvent of extract and get the flavonoidal content.
ARELLANO UNIVERSITY - COLLEGE OF PHARMACY
PAGE
20
Determination of % Yield of Crude Extract
After the crude extract was concentrated into
a syrupy
concentration, percentage yield was determined using the formula:
%Yield = weightweight ofof thethe crudedried sampleextract x100
Physicochemical Test
Physical Test: Determination of physical test according to color , odor, and appearance.
Solubility Test: The solubility in the extract in water and in different organic solvents was determined
in the following procedures as specified in the United States Pharmacopeia (USP). Measure 1g of the
petroleum ether extract of Citrofortunella microcarpa and determine its solubility of each of the following
solvent: Ethanol, Petroleum ether, Chloroform, Acetone and Distilled water.
Table 1. Solubility Testing According to USP Standard.
Descriptive Term
Parts of Solvent Required for 1 Part of
Solute
Very Soluble
Less than 1
Freely Soluble
From 1 to 10
Soluble
From 10 to 30
Sparingly Soluble
From 30 to 100
Slightly Soluble
From 100 to 1,000
Very Slightly Soluble
From 1,000 to 10,000
Practically insoluble or
Greater than or equal to 10,000
Insoluble
Confirmatory test
To determine the flavonoidal content of Citrofortunella microcarpa using the different

reagents, the researcher preformed the following tests:
For Alkaline Reagent Test, petroleum ether extract was treated with few drops of
sodium hydroxide solution and the mixture created a formation of intense yellow color.
On further addition of dilute acid, colorless thus indicating the presence of flavonoids.
For the Lead acetate test, petroleum ether extract was treated with few drops of lead
acetate solution, and the mixture of yellow precipitate – again indicating the presence
of flavanoids.
Lastly, for Ferric Chloride test, petroleum ether extract was treated with a few drops of
ferric chloride forms green color which also indicates the presence of flavonoids.
Reagents and Materials
All reagent tests solutions were provided by the College of Pharmacy laboratory stock
rooms. Metformin hydrochloride were obtained from Animal Life Veterinary Clinic
located in Burgos St., Quezon, Solano, Nueva Vizcaya. The alloxan monohydrate were
purchased from Chemline Scientific Corporation in Novaliches, Quezon City. Normal
Saline solution was purchased from Watson’s Philippines, Savemore Mezza Residences.
Experimental Animals
Twenty (20) Sprague Dawley rats were purchased from the Biology Department and
Animal Care Center of the University of the Philippines - Diliman. Random sexes of
rats were used aging from 10-12 weeks old. Four groups of five (5) rats were prepared
and labeled as follows: Group A Negative control; Group B 250 mg/Kg of Citrofortunella
microcarpa peel extract; Group C 500 mg/Kg of Citrofortunella microcarpa peel extract
and Group D 500mg/Kg Metformin HCl, the standard drug.
Rats were kept for seven days for acclimatization. Given sufficient supply of water and
the lights were constantly being turned on and off every
12 hours. After the seventh -day acclimatization, initial blood sugar level was obtained
to determine the normal glucose level of the test animals.
Induction of Hyperglycemia
After the baseline glucose levels were obtained, test animals were given 2.5g/kg
Alloxan monohydrate intraperitonially to induce hyperglycemia. Thirty minutes after
the induction, blood glucose levels were measured.
Biological Testing
During the treatment, the rats were fed on a normal diet and they were given distilled
water ad libitum; Group B and C were the treatment groups and received 250 mg/kg
and 500 mg/kg of Citrofortunella microcarpapeel extract respectively. Group D which
served as the positive control were given 500 mg/kg Metformin Hydrochloride. On day
0, 4th, 8th and 15th day, blood samples were obtained and blood sugar levels were
recorded.
Statistical analysis
Oneway ANOVA was conducted to test the statistical significance of the results. It is
applied for samples with more than two populations to test whether the are differences
between the means of
several groups, based on samples taken from each population. It is also used to
compare the means of 3 or more populations in the study.
RESULTS AND DISCUSSION
Organoleptic evaluation refers to means of the organs of senses and includes

appearance, color, odour and senses
Table 2. Results of Solubility Test
SOLVENT
Expected Result
Ethyl Alcohol
Soluble
Petroleum ether
Soluble
Chloroform
Soluble
Acetone
Soluble
Distilled Water
Insoluble
The Table 2 shows the solubility of the petroleum ether extract of Citrofortunella
microcarpa peels in different solvents with varying polarity. As the result indicates,
Citrofortunella microcarpa is soluble in ethyl alcohol, petroleum ether, chloroform, and
acetone while insoluble in distilled water.
PAGE
25
Table 3. Results in Organoleptic Evaluation
Color
Dark green in color

Odor
Pungent
Taste
Sour taste
Appearance
Semi-solid
As for the result of the organoleptic test presented in Table 3, the extract turned to dark green color,
sour taste, semi-solid in appearance and has pungent odor.
Table 4. Percentage Yield
TRIAL 1
TRIAL 2
Weight of Citrofortunella
500 g
500 g
microcarpa peel
Weight of Empty Evaporating
47.83 g
51.42 g
Dish
Weight of Evaporating Dish
61.18 g
63.27 g
with Citrofortunella
microcarpa peel extract

Weight of Citrofortunella
13.35 g
11.85 g
microcarpa peel extract
Percentage Yield
2.67 %
2.37 %
AVERAGE
2.52
The chart shows the result of percentage yield two trials have been done for trial 1 the total percentage
yield is 2.67%, for trial 2 2.37% and has an average percentage yield of 2.52%.
Table 5. Confirmatory Test for Flavonoids

For the chemical test in Alkaline reagent, the actual results has shown a formation of
yellow solution which indicates the presence of flavonoid. In Lead acetate, the actual
result has shown a formation of yellow precipitate which indicates the presence of
flavonoids. In Ferric Chloride, the actual result has shown a formation of green
coloration indicating the presence of flavonoids.
Table 6. Mean Blood Glucose Levels in the Different Groups of
Rats at different Days
Day
Group A
Group B
Group C
Group D
Alloxan +
(Alloxan +
(Alloxan +
(Alloxan + 500
NSS
250mg/kg
500 mg
mg/kg
extract)
extract)
Metformin HCl)
After
6.2
6.1
5.5
5.6
Induction
0 Day
6.1
5.8
5.3
5.4
4th Day
5.8
5.6
5.3
5.1
8th Day
5.5
5.3
3.8
4.9
15th Day
3.8
4.1
4.9
5.3
7
6
After Induction 0 Day 4th Day 8th Day 15th Day
Group A Alloxan + NSS
Group B (Alloxan + 250mg/kg extract)
Group C (Alloxan + 500 mg extract)
Group D (Alloxan + 500 mg/kg Metformin HCl)

Figure 5. Mean Blood Glucose Levels in the Different Groups of
Rats at different Days

The treatment procedures that have been conducted for 15 days in Table 6, starting
after induction, a change in glucose level of the rats was already observed. For group A
as the negative control there was a change of glucose level after induction to 0 day with
the initial decreased of 0.1mmol/L, on 4th day it decreased to 0.3mmol/L, on the8th day
it decreased 0.3mmol/L, on the 15th day decreased to 1.7mmol/L. For Group B which
was given 250mg/kg extract after induction to 0 day it decreased 0.3mmol/L, 4 th day
decreased 0.2mmol/, 8th day 0.3 mmol/L, 15th day it decreased 1.2mmol/L. For Group
C which was given 500mg/kg extract after induction to 0 day it decreased up to
0.2mmol/L, 4th day no changes in blood glucoses level. 8th day 1.5mmol/L, 15th day it
increased to 1.1mmol/L. For Group D as the positive control, after induction to 0day
initial level decreased at 0.2mmol/L, 4th day it decreased 1.3mmol/L, 8th day it
decreased 0.2mmol/L, 15th day it increased 0.4mmol/L.
Table7. Percent Decline in Blood glucose level
(based on previous reading)
Day
Group A
Group B
Group C
Group D
Alloxan +
(Alloxan +
(Alloxan +
(Alloxan + 500
NSS
250mg/kg
500 mg
mg/kg
extract)
extract)
Metformin
HCl)
After
1.61%
4.91%
3.63%
3.57%
induction
vs. Day 0
Day 0 vs.
4.91%
3.44%
5.56%
Day 4
Day 4 vs.
5.17%
5.36%
28.30%
3.92%
Day 8
Day 8 vs.
30.90%
22.64%
-28.94%
-8.16%
Day 15
40.00%
30.00%
20.00%
10.00%
0.00%
Group A
Group B
Group C
Group D
-10.00%
-20.00%
-30.00%
-40.00%
After induction vs. Day 0
Day 0 vs. Day 4

Day 4 vs. Day 8
Day 8 vs. Day 15

Figure 6. Percent Decline in Blood glucose level
(based on previous reading)

Table 7 showed different blood glucose levels for the different changes in day to day
treatment were computed according to their percentage declined from after induction
day up to day 15. For Group A as the Alloxan + NSS after induction to Day 0 a change
of glucose of 1.61%, Day 0 to Day 4 with a percent decline of 4.91%, Day 4 to Day 8 a
percent decline of 5.17%, Day 8 to Day 15 has a percent decline of 30.90%. And for the
Group B as the 250 mg/kg extract after induction to Day 0 has a percent decline of
4.91%, Day 0 to day 4 has 3.44% in percent decline while in Day 4 to Day 8 a change
of 5.36% and Day 8 to Day 15 has a 22.64%. For Group C as the 500mg/kg extract
after induction to Day 0 has a 3.63% decline compared to day 0 to Day 4 there was no
percentage decline or any changes according to these respective days and Day 4 to Day
8 28.30% was declined and for the Day 8 to Day 15 -28.94% was declined means it
posessed a hypoglycemic activity in this dose. And lastly for group D after induction to
day 0 3.57% was declined, for day 0 to day 4 5.56%. Day 4 to Day 8 3.92% and for 8
day up to 15 day -816% was declined means there is a high decreased in blood glucose
level.
Statistical Analysis
The significance of differences between the mean of the treated and control value must
be less than 0.05 to be considered significant.
PAGE
31
Table8. Comparison of Alloxan+NSS in Day to day Treatment
After Induction
Day 0
0.9994
Not Significant
Day 0
Day 4
0.9714
Not Significant
Day 4
Day 8
0.9929
Not Significant
Day 8
Day 15
0.0209
Significant
Table 8 showed that Alloxan in NSS in Day 8 to Day 15 which has a value of 0.0209 has significant
difference compared to different days. It exhibited hypoglycemic effect and considered to be effective.
Table 9. Comparison of Alloxan + 250mg/kg extract in respective days

After Induction
Day 0
0.9973
Not Significant
Day 0
Day 4
Not Significant
Day 4
Day 8
0.9959
Not Significant
Day 8
Day 15
0.794
Not Significant
Table 9 shows that Alloxan+250 mg/kg extract in different respective days did not shows significant
differences as mention above all did not show hypoglycemic effect means the dose of 250/kg extract is
not that as effective.
Table 10. Comparison of Alloxan+500 mg/kg extract in respective days
After Induction
Day 0
0.9991
Not Significant
Day 0
Day 4
Not Significant
Day 4
Day 8
0.9489
Not Significant
Day 8
Day 15
Not Significant
Table 10 shows in different respective days did not shows significant differences as
mention above all did not show hypoglycemic effect means the dose of 500/kg extract
is not that as effective.
Table 11. Comparison of Alloxan+500 mg/kg Metformin HCL in respective days
After Induction
Day 0
0.9416
Not Significant
Day 0
Day 4
0.9967
Not Significant
Day 4
Day 8
0.9999
Not Significant
Day 8
Day 15
0.995
Not Significant
Table 11 showed that Alloxan+500 mg/kg Metformin HCL has no significant
differences in comparison in respective days. The values of mean that among the 2
different dose levels has the same results to standard drug Metformin HCl. It did not
reach the required values to considered comparable.
CONCLUSION
Paired difference test was used to compare the mean of the baseline and final blood
glucose levels. Post-Hoc analysis Dunnett’s comparison testing was used to determine
if there is a significant difference between groups. It can be concluded that there is no
significant difference(p>0.05) between the effectiveness of Citrofortunella microcarpa
extracts 250mg/KBW and 500mg/KBW and the standard drug Metformin HCl. The
data presented in this research proves that Citrofortunella microcarpa possesses a
property that can potentially lower blood glucose level and the dosage of 500mg/kg
was found to be more effective as compared to 250 mg/kg. Therefore, the researcher
claims that petroleum ether extract of Citrofortunella microcarpafruit peel has a glucose
lowering property that could potentially manage, if not cure, diabetes.
RECOMMENDATION
The results of the study could be further explored by the following recommendations:
Use other solvents and methods of extraction in order to maximize the full potential of
Citrofortunella microcarpa as a glucose lowering agent;
Use instruments such as HPLC, GC-MS, NMR and other apparatus to elucidate the
specific flavonoid responsible for the glucose lowering property of Citrofortunella
microcarpa fruit peel;
Conduct toxicity test on the samples to be used;
Formulate a suitable pharmaceutical dosage form that is economically feasible;
Make a further study on the hypoglycemic and antidiabetic potential of other parts of
the plant Citrofortunella microcarpa; and
Histophatologically evaluate the pancreas of the experimental
animals.
APPENDICES
Day to day Activity
Collection of Citrofortunella microcarpa
II
Authentication
III
Extraction
IV
Physicochemical testing
Acclimatization of rats
12hours dark
12hours light
Normal animal pellet

Distilled water was given ad libtium
VI
Obtaining 1 drop of blood for the normal glucose level using
glucometer
VII
Inducing alloxan monohydrate and getting the initial levels
increase in blood sugar level and preparing for the treatment
(0th day) treated with extracts and standard drug before

obtaining blood glucose level
VIII
Treatment and normal diet
IX
Treatment and Glucose level obtained (4th day)
Treatment
XI

XII
Treatment
XIII

Group C
Weight
Rat 1
99.2 grams
Rat 2
102.3 grams
Rat 3
100.5 grams
Rat 4
100.7 grams
Rat 5
97.6 grams
Average= 100.6 grams
Group D
Weight
Rat 1
98.8 grams
Rat 2
99.1 grams
Rat 3
108.7 grams
Rat 4
102.4 grams
Rat 5
105.2 grams
Average= 102.84 grams

PAGE
37
Normal Blood glucose Level before Alloxan induction

GROUP A
GROUP B
GROUP C
GROUP D
Rat 1= 3.3mmol/L
Rat 1= 3.2mmol/L
Rat 1= 5.0mmol/L
Rat 1= 4.7mmol/L
Rat 2= 4.6mmol/L
Rat 2= 3.8mmol/L
Rat 2= 4.7mmol/L
Rat 2= 3.6mmol/L
Rat 3= 3.9mmol/L
Rat 3= 4.1mmol/L
Rat 3= 5.3mmol/L
Rat 3= 3.3mmol/L
Rat 4= 4.1mmol/L
Rat 4= 5.0mmol/L
Rat 4=6.3mmol/L
Rat 4=3.3mmol/L
Rat 5= 5.0mmol/L
Rat 5= 3.8mmo/L
Rat 5= 4.0mmol/L
Rat 5=4.1mmol/L
Ave= 4.2mmol/L
Ave= 3.9mmol/L
Ave= 5.4mmol/L
Ave= 3.1mmol/L
Blood Glucose level each Rat in day-day treatment
Day
Group A
Group B
Group C
Group D
Alloxan + NSS
(Alloxan +
(Alloxan + 500 mg
(Alloxan+500mg/kg
250mg/kg extract)
extract)
Metformin HCl)
After
Rat 1= 7.2mmol/L
Rat 1= 3.3mmol/L
Rat 1= 5.9mmol/L
Rat 1= 7.7mmol/L
Induction
Rat 2= 6.6mmol/L
Rat 2= 5.9mmol/L
Rat 2= 4.9mmol/L
Rat 2= 4.3mmol/L
Rat 3= 5.8mmol/L
Rat 3= 8.0mmol/L
Rat 3= 6.0mmol/L
Rat 3= 3.9mmol/L
Rat 4= 4.9mmol/L
Rat 4= 5.2mmol/L
Rat 4=6.4mmol/L
Rat 4=6.0mmol/L
Rat 5=6.6mmol/L
Rat 5= 8.1mmo/L
Rat 5= 4.5mmol/L
Rat 5=9.0mmol/L
Average:6.2
Average:6.1
Average: 5.5
Average: 5.6
0 Day
Rat 1= 7mmol/L
Rat 1= 3.3mmol/L
Rat 1= 5.7mmol/L
Rat 1= 4.7mmol/L
Rat 2= 6.5mmol/L
Rat 2= 5.1mmol/L
Rat 2=4.7mmol/L
Rat 2= 4.5mmol/L
Rat 3= 5.7mmol/L
Rat 3= 7.1mmol/L
Rat 3=5.7mmol/L
Rat 3= 3.9mmol/L
Rat 4= 4.7mmol/L
Rat 4= 5.3mmol/L
Rat 4=6.4mmol/L
Rat 4= 6.1mmol/L
Rat 5= 6.6mmol/L
Rat 5= 8mmol/L
Rat 5= 4.4mmol/L
Rat 5= 8mmol/L
Average: 6.1
Average:5.8
Average:5.3
Average:5.4
th
4 Day
Rat 1= 6.5mmol/L
Rat 1= 3.3 mmol/L
Rat 1=5.8 mmol/L
Rat 1= 4.5 mmol/L
Rat 2= 6.2mmol/L
Rat 2=5.1mmol/L
Rat 2= 4.6 mmol/L
Rat 2= 4.5 mmol/L

Rat 3= 5.4mmol/L
Rat3= 7mmol/L
Rat 3= 5.7mmol/L
Rat 3= 3.5 mmol/L
Rat 4= 4.6mmol/L
Rat 4=5 mmol/L
Rat 4= 6.1 mmol/L
Rat 4= 6 mmol/L
Rat 5= 6.2mmol/L
Rat 5=7.8mmol/L
Rat 5= 4.4 mmol/L
Rat5= 7 mmol/L
Average: 5.8
Average: 5.6
Average: 5.3
Average:5.1
8th Day
Rat 1= 6.2mmol/L
Rat 1= 3 mmol/L
Rat 1= 5.2mmol/L
Rat 1= 4.5mmol/L
Rat 2= 6 mmol/L
Rat 2= 5 mmol/L
Rat 2= 4mmol/L
Rat 2= 4.6mmol/L
Rat 3= 5.1mmol/L
Rat 3= 6.6mmol/L
Rat 3= 5mmol/L
Rat 3= 3.2 mmol/L
Rat 4= 4.5mmol/L
Rat 4= 4.7mmol/L
Rat 4= 5.8mmol/L
Rat 4= 5.8 mmol/L
Rat 5= 6mmol/L
Rat 5= 7 mmol/L
Rat 5= 4.3mmol/L
Rat 5=6.7 mmol/L

Average:5.5
Average: 5.3
Average: 3.8
Average:4.9
15th Day
Rat 1= 3.3 mmol/L
Rat 1= 4.6 mmol/L
Rat 1= 3.3mmol/L
Rat 1= 6.6mmol/L
Rat 2= 4.2mmol/L
Rat 2= 3.6 mmol/L
Rat 2= 4.4mmol/L
Rat 2= 4.8mmol/L
Rat 3= 3.3mmol/L
Rat 3= 4mmol/L
Rat 3= 6.8mmol/L
Rat 3= 4.1 mmol/L

Rat 4= 5mmol/L
Rat 4= 3.8mmol/L
Rat 4= 6.4 mmol/L
Rat 4= 5.2 mmol/L
Rat 5= 3.1mmol/L
Rat 5= 4.5 mmol/L
Rat 5= 3.5mmol/L
Rat 5= 6mmol/L
Average: 3.8
Average: 4.1
Average= 4.9
Average: 5.3
PAGE
38
Amount and volume of Alloxan administered based on their body
weight
Weight(g)
Amount of alloxan
Volume (mL)
(mg)
Rat 1
100.4 grams
15.06mg
0.60 mL
Rat 2
98.2 grams
14.73 mg
0.59 mL
Rat 3
100.1 grams
15.02 mg
0.60 mL
Rat 4
97.5 grams
14.63mg
0.59 mL
Rat 5
100.8 grams
15.12mg
0.60 mL
Average
99.4 grams
14.912mg
0.60 mL
Group B
Weight(g)
Amount of alloxan
Volume (mL)
(mg)
Rat 1
101.9 grams
15.29 mg
0.61mL
Rat 2
100.2 grams
15.03 mg
0.60mL
Rat 3
111.6 grams
16.74 mg
0.67 mL
Rat 4
103.8 grams
15.57 mg
0.62mL
Rat 5
102.7 grams
15.41 mg
0.62mL
Average
104.4 grams
15.61mg
0.62mL
Group C
Weight(g)
Amount of
Volume
alloxan (mg)
(mL)
Rat 1
99.2 grams
14.88 mg
0.60 mL
Rat 2
102.3 grams
15.35 mg
0.61 mL
Rat 3
100.5 grams
15.08 mg
0.60 mL
Rat 4
100.7 grams
15.11 mg
0.60 mL
Rat 5
97.6 grams
14.6 mg
0.59 mL
Average
100.6 grams
15.01 mg
0.6mL
Group D
Weight(g)
Amount of alloxan
Volume (mL)
(mg)
Rat 1
98.8 grams
14.82 mg
0.59 mL
Rat 2
99.1 grams
14.87 mg
0.59mL
Rat 3
108.7 grams
16.31 mg
0.65 mL
Rat 4
102.4 grams
15.36 mg
0.61 mL
Rat 5
105.2 grams
15.78 mg
0.63 mL
Average
102.84 grams
15.43 mg
0.61 mL
Amount and volume of Metformin HCl to administer in Positive control based on

their body weight
500 mg dissolve in 10mL aqueous water
Group D
Weight(g)
Amount of
Volume (mL)
metformin (mg)
Rat 1
98.8 grams
49.4mg
0.99mL
Rat 2
99.1 grams
49.55mg
0.99mL
Rat 3
108.7 grams
54.35mg
1.09mL
Rat 4
102.4 grams
51.2mg
1.024mL
Rat 5
105.2 grams
52.6mg
1.052mL
Average
102.84 grams
51.42mg
1.03mL
Computation for getting the %mean difference of the decreasing initial values day to day
Formula:
%decline in glucose after induction to 0 day
=
−0
x 100
%decline in 0 day to 4th day
=
0
x100
−4 ℎ
0
%decline in 4th day to 8th day = −

4 ℎ
8 ℎ−
15 ℎ
15 100
ℎ
%decline in 8th day to 15th day = 100

Group A: Alloxan+ NSS 6.2−6.1
6.2
x 100 = 1.61%
after induction to 0 day
0 day to 4th day

6.1
4thday to 8th day
5.8
3.8
8th day to 15 day

Group B: Alloxan+ 250mg/kg
x 100
=
6.1
6.1−5.8
= 4.91%
5.8
x 100 =
0 day to 4th day

5.8−5.6
x 100
3.44%
5.6
4thday to 8th day
5.6−5.3
= 5.36%
8th day to 15 day
5.3
Group C: Alloxan + 500mg/kg
5.5
x 100
5.5−5.3
= 3.63%
5.3
x 100 =
0 day to 4th day
5.3−5.3
x 100
0%
=
5.3
4thday to 8th day
5.3−3.8
= 28.30%
3.8
x 100 =
8th day to 15 day
3.8−4.9
-28.94%
PAGE
41
Group D: Alloxan + 500mg/kg
=
5.6
x 100
5.6−5.4
= 3.57%
5.4
x 100 =
0 day to 4th day
5.4−5.1
x 100
5.56%
5.1
4thday to 8th day

5.1−4.9
= 3.92%
5.3
x 100 =
8th day to 15 day
4.9−5.3
-8.16%
Confirmatory Test
FerricChloride Test
(green coloration)
Alkaline Reagent Test
(yellow coloration)
LeadAcetate Test (yellow precipitate)

Solubility testing
Ethanol
Chloroform
Acetone
(slightly soluble)
(soluble)
Petroleum Ether
Water
(soluble)
Materials and Methods
Steam
Rotary
evaporator
Flavonoid content
after steam bath

Dried Citrofortunella
Microcarpa
Oral gavage
Maceration
Inducing of Alloxan
Blood Collection
Intraperitoneal
Acclimatization
AUTHENTICATION
PURCHASING
CURRICULUM VITAE
Cuntapay, Geneece O.
Address: Tower 2 3702 Mezza Residences Quezon
City, Manila
Tel./cellphone number: 09361874381
E-mail address: geneece_c@yahoo.com
Birthday: November 26, 1996
EDUCATION
Primary
Dupax del Norte Central School

2003-2009
Dupax del Norte Nueva Vizcaya
Secondary
Saint Mary’s School of Dupax
2009-2013
Dupax del Sur Nueva Vizcaya
Tertiary
Centro Escolar University Manila
San Miguel St. Mendiola Manila
2013-2017
ACTIVITIES AND ACHIEVEMENTS
4th Honorable mention S.Y 2003-2009
8th Honor S.Y 2009-2013

Member
Junior Pharmacist Association

ROVILYN DAYAG DELA CRUZ
42 Panlingalingan St., Domang,
Dupax del Sur, Nueva Vizcaya
Contact No: 0915-4925-593
E-mail: Delacruzrovilyn48@gmail.com
LICENSE/CERTIFICATE
Professional Teacher License (August 2015)
TESOL Certificate (2014)
120-hour TEFL certificate (January 2016)
EDUCATIONAL BACKGROUND:
Tertiary Philippine Normal University - Manila

Taft Avenue, Manila
BSE English
June 2011 – March 2015
Honors’ Class
Secondary Saint Mary’s Dupax
Honorable Mention
Primary Dupax Central School

Honorable Mention
PROFESSIONAL SKILLS:
Good understanding of all teaching methods and learning styles
Able to convey materials to students in an easy and understandable manner
Strong teaching presentation, verbal and written communication
Open to criticisms and suggestions for self-improvement
Possesses good Scholastic Record RELEVANT
EXPERIENCES:
ENGLISH INSTRUCTOR at HITUTOR (July 2016-December 2017)
Conducted English classes with Taiwanese students based on the given textbook
appropriate to their level.
Constructed relevant questions that engage students in meaningful discussions.

Noted student’s errors and put it on student’s documentation for review.
ENGLISH INSTRUCTOR at EFLG, Inc. (December 2015-July 2016)
Conducted English classes with Korean students.
Constructed relevant questions that engage students in meaningful discussions.
Corrected students’ errors by the end of the class.
Submitted daily news articles which are being put in the textbook and being used in
daily discussions
Performed Level Test to gauge students’ English fluency and assign appropriate
textbook.
PRACTICE TEACHING at Magsaysay High School, Espana, Sampaloc Manila
Planned daily lesson and activities for Grade 7 students during the first quarter of S.Y.
2014-2015.
Motivated the students to participate in class discussion through interactive activities

and active discussion.
Resolved class conflicts through open house confessions.
Graded students’ homework, outputs, and class participation.
FIELD STUDY at PNU, Institute for Teaching and Learning (ITL)
Observed school and classroom set-up that affects the quality of learning of students.
Conducted case study about certain students and resolved problematic aspects.
Planned and demonstrated a writing lesson.

SEMINARS ATTENDED
•International NeuroELT Conference: “Innovationas in
Language Teaching Psychology, Education and Neuroscience” at Philippine Normal

University (January 22-
24, 2015)
ELT and the Science of Happiness and DIY: NeuroELT - Making Your Class More
Brain-friendly at Philippine Normal University (October 8, 2014)
English Fortnight and Reading: “My Life as a Woman Writer” at Philippine Normal
University (November 27, 2014)
BOS: “Basic Orientation Seminar-Workshop on Debate” at Philippine Normal

University (November 19, 2014)
•Asian International TESOL Conference; Global Trends In Language Teaching And

Learning at Henry Sy Building, De La Salle University –Manila (January 30 – February
2, 2014)
SMI-IC Ethics Bowl at St. Scholastica’s College (February 15, 2013)
E-lecture Series 1: Broadcast Journalism and Newspaper Writing at Philippine Normal

University (July 25, 2012)
•E-lecture Series 3: A Lecture Forum on 21st Century Skills: A Challenge to the

English Teachers at Philippine
Normal University ( November 15, 2012)

AFFILIATION
HONORS’ CLASS COMMITTEE MEMBER at Philippine Normal University, S.Y. 2012-

2015
ENGLISH CLUB MEMBER at Philippine Normal University,
S.Y. 2012-2015
DABATE MEMBER at Philippine Normal University, S.Y.

2012-2013
•PIO OF THE STUDENT SUPREME GOVERNMENT at Saint Mary’s Dupax, S.Y. 2010-
2011
•CLASS PRESIDENT at Saint Mary’s Dupax, S.Y. 2008-2009
PERSONAL INFORMATIONS:
Age:22
Civil Status: Single
Gender: Female
Nationality:Filipino
Father: Romeo B. Dela Cruz

Occupation:Teaching
Mother:Vicenta D. Dela Cruz
Occupation: Teaching

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ARELLANO UNIVERSITY - COLLEGE OF PHARMACY PAGE 1

GLUCOSE-LOWERING PROPERTY OF Citrofortunella microcarpa

FRUIT PEEL USING ALLOXAN-INDUCED HYPERGYCEMIC RATS

Ms. Regine Gonzales, RPh

Arellano University – College of Pharmacy

Keywords: glucose-lowering, alloxan,Citrofortunella microcarpa, hyperglycemia,

date, there is noreport of phenolic acids content in locally available Citrofortunella

Determine the solubility of Citrofortunella microcarpa peel extract.

Determine the concentration of Citrofortunella microcarpa peel extract exhibited

STATEMENT OF THE PROBLEM

What is the percentage yield of Citrofortunella microcarpa?

Is flavonoid present in Citrofortunella microcarpa peel extract?

What concentration of Citrofortunella microcarpa peel extract exhibits a hypoglycemic

Is there a significant difference between the hypoglycemic activity of Citrofortunella

REVIEW OF THE RELATED LITERATURE

The botanical classification of Anamirta cocculus is classified under family

Subkingdom Tracheobionta – Vascular plants Superdivision Spermatophyta – Seed plants

Division Magnoliophyta – Flowering plants

Class Magnoliopsida – Dicotyledons

Subclass Rosidae Order

Family Rutaceae– Rue family

Citrofortunella microcarpa is a smooth and slightly spiny plant, growing to a height of 3

Figure 1. Citrofortunella microcarpa (Calamansi)

In Higanoon, a tribe of Mindanao use decoction of leaves to lower hypertension. Juice

In Malaya, combined with pepper to help expel phlegm. Root

A study on the constituents of Citrofortunella microcarpa or whole fruit essential oil

Citrofortunella microcarpa has been demonstrated to exhibit antioxidant function and

Hepatoprotective, peel extract exhibited hepatoprotective activity against

maceration with water. Anti-inflammatory property is effective in increasing dose of

Diabetes a condition where the pancreas produce insufficient amount of insulin or

commonly in people of First Nations descent, Hispanics, and North Americans of

Causes of Type I Diabetes

Causes of Prediabetes and Type II Diabetes

Sign and Symptoms

Diagnosis and Treatment

Alloxan monohydrate – Inducer

( Depomed Inc. et,. Al 2016)

Figure 2. Alloxan Monohydrate Chemical Structure, (Pubchem, 2005)

Metformin Hydrochloride - Standard Drug

Metformin is an antihyperglycemic agent, which improves glucose tolerance in patients

Figure 3. Metformin Chemical Structure, (Pubchem, 2005)

Citrofortunella microcarpa or better known as Calamansi/Kalamansi is a commercially

Citrofortunella microcarpa also contains lignin, a fiber-like component. This tends to

Citrtrofortunella microcarpa or Citrus mitis, also known as 'limaukasturi' is an edible

Collection of Citrofortunella microcarpa

Authentication, Pre paration and Drying

of the f ruit peel

Petroleum Ether Extractives

Lead acet ate test

Ferric chl oride test

Acclimatization of experimental animals

Induction of Hype rglycemia

Testing of Pharma cological effect

Figure 4. Paradigm of the Study

To determine if Citrofortunella microcarpa, L. fruit peel has a glucose lowering property,

Collection and Preparation of Plant Material

Determination of % Yield of Crude Extract

After the crude extract was concentrated into

concentration, percentage yield was determined using the formula:

%Yield = weightweight ofof thethe crudedried sampleextract x100

Table 1. Solubility Testing According to USP Standard.

Parts of Solvent Required for 1 Part of

From 100 to 1,000