The Potenial of Eleanor Roosevelt Croton (Codiaeum variegatum) Leaves Extract
as Blood Anticoagulant
Shiela V. Pilapil
Chloe Marie S. Urot
Sharmaine B. Nono
ABSTRACT
This study aimed to explore the potential
of C.variegatum as blood anticoagulant.
The C.variegatum leaves were air-dried
and filtered after it was soaked in
ethanol for 48 hours and after that,
crude plant extract was obtained
through modified oil bath technique. The
effectivity of 100% concentration of plant
extract was tested to 50µl of blood
plasma using Activated Partial
Thromboplastin Time Test (APTT)
together with the control in three trials.
Result showed that C.variegatum
effectively delayed the coagulation of
blood. Therefore, there is no significant
difference in the mean coagulation rate
of blood plasma in different age bracket
of male and female blood samples. The
researchers conclude that C.
variegatum has a potential to be a blood
anticoagulant drug because of the
compound flavonoids and alkaloids
present in this plant.
Keywords: Anticoagulant, Activated
Partial Thromboplastin Time Test
INTRODUCTION
For the past 15 years, heart
disease continues to be the top cause of
death among the Filipinos. In 2016, an
average of 1,591 persons died daily.
This translates to 66 deaths per hour or
one (1) per minute. The number of
deaths from 2006 to 2016 showed an
increasing trend. The increase during
the ten-year period is almost one-third,
or 32.0 percent, from 441,036 in 2006 to
582,183 in 2016. The reason for the
increase may have been due to
ischaemic heart disease that was the
top leading cause of death with 74, 134
or 12.7 percent of the total. (PSA, 2020)
A recent data showed a staggering
122,950 deaths and a death rate of
191.79 per 100,000 of population, in
which the Philippines took the number
28 rank of the highest heart disease
death in the world. (WHO, 2017).
Platelet activation in
atherosclerotic arteries is central to the
development of arterial thrombosis;
therefore, a precise control of platelet
function is imperative in preventing
thrombotic events (Jungln, 2017).
Thrombotic disorders increases the risk
of cardiovascular diseases and
represents a major health problem in
world wide. Those anticoagulants which
block the clotting factors or coagulation
factors and in turn inhibit the clot
formation are extensively used in the
treatment of thrombotic disorder. The
thrombotic disorders includes arterial
thrombosis, atrial fibrillation, myocardial
infarction, unstable angina, deep vein
thrombosis, pulmonary embolism and
cerebral stroke. Thus, anticoagulants
play an effective role in the prevention
and treatment of thrombotic disorders
(Ashwini, 2017). Blood thinners is also
known as anticoagulants and antiplatelet
agents but they can’t thin the blood.
 They are used to reduce
excessive blood clot formation. Too
much of clotting of blood can cause
thrombus which can stop the normal
circulation of blood flow. This is known
as thrombosis. Thrombus travel to larger
blood vessels and block the smaller
blood vessels (Kelson, 2015). Common
anticoagulants include warfarin and
heparin. The usage of anticoagulants in
modern medicine is vitally important and
needed. Patients suffering from recent
heart attacks, with atrial fibrillation
(irregular heartbeat), with artificial heart
valves, or those who have heart disease
such as cardiomyopathy benefits from
taking these anticoagulant drugs.
Recent medical developments have
made the anticoagulant drugs very
much accessible and available to the
wide consumers, one example of it is
the drug warfarin. Warfarin works by
slowing the production of some of the
chemicals in your body that are needed
for blood clotting. It is the only and old
drug that has been used for 50 years
with proven; highly effective if used
correctly. However, warfarin is
expensive and may not be readily
available in drug stores especially in the
far flung areas and it also Warfarin is an
anticoagulant medicine (blood thinner).
Before the discovery of the new
anticoagulant drugs, warfarin is the first
drugs that was widely used to treat and
to prevent the thromboembolic diseases
that caused by the abnormal coagulation
of blood inside the body .In the patients
who have liver disease or renal
insufficiency, New Oral Anticoagulant
(NOACs) is not advisable to be taken.
Even though warfarin has a
disadvantage, many people use it
because of its affordable compared to
commercials anti-coagulant drugs.
Thrombus happens mostly in obese
patients and premenopausal women at
the age of 50 to 70 years old (Ritva,
2016).
Due to the rising cases of heart
disease in the country, the need of an
anticoagulant drug that is cheap and
readily available to the patients is of
utmost need. The researchers studied
the anticoagulant properties of the
Eleanor Roosevelt croton (Codiaeum
variegatum) leaf extract and its potential
as a substitute natural anticoagulant
drug that can be made available to the
especially to the people living in far flung
areas in which the access to the
market’s available anticoagulant drugs is
still an issue. Moreover, the usage of the
plants anticoagulant properties could
potentially minimizes or eliminates the
risk of the side effects caused by the
market’s available blood anticoagulant
drugs.
The search of an alternative
anticoagulant that is at least as effective
or much better surpassing it while
eliminating most of its serious side
effects formulated the idea of identifying
the potential anticoagulant properties of
Eleanor Roosevelt croton (Codiaeum
variegatum) leaf extract. The plant is
readily accessible, easily used and
prepared and can be obtained in the
locality making it cheap and efficient
substitute to the anticoagulant drugs
available in the market. The Philippines
being one of the top countries in the
world suffering from heart disease could
benefit hugely from developing natural
anticoagulant drugs that can be readily
made and cheap. The study of the
Eleanor Roosevelt croton (Codiaeum
variegatum) leaf extract anticoagulant
properties could help in developing an
alternative natural heart disease
medicine. The notable problems of using
the available anticoagulant drugs in the
market is that it is expensive, needs
sample knowledge in dosage and
monitoring and may not be readily
accessible to far flung areas. Also, the
study of the Eleanor Roosevelt croton
leaf extract as an anticoagulant might
shift the demand of the patients from
using the market’s anticoagulant drugs,
which has notable and serious side
effects, to a natural substitute which
may minimize the said side effects.
Eleanor Roosevelt leaf extract
anticoagulant activities were observed
and recorded. In conclusion, the plant
extract did exhibit anticoagulant
properties and therefore could be
potentially studied and developed as a
natural and cheaper alternative to
modern anticoagulant drugs. Most of the
people do not have the ability to buy
drugs in the store that can help them to
ease their risk suffering diseases cause
by blood anticoagulant. This study aims
to find a new organic plant that is a
better source of blood anticoagulant that
anyone of us can afford and has the
ability to give cure on the said diseases.
This study is also expected to decrease
human blood coagulation rate.
MATERIALS AND METHODS
The experimentation was
conducted with three phases: Phase I:
Preparation of Materials; Phase II:
Experimentation; Phase III: Data
Collection and Analysis. The
Preparation of materials and plant
samples are assembled in the laboratory
of Nabunturan National Comprehensive
High School and the collection of blood
samples and testing the plant extract
was done in Mawab, Tagum City
laboratory. The results were computed
and analyzed.
Phase I. Preparation of Materials
Collection of Plant Samples
Eleanor Roosevelt Croton leaves
were collected from P-3, Linda
Nabunturan, Davao de Oro. The
researcher were gathered for about 1kg
of Eleanor Roosevelt Croton leaves.
Preparation of Plant Samples
The collected Eleanor Roosevelt
croton leaves were washed thoroughly
using running water. The mass of the
leaves was recorded first before
washing. The leaves were air dried at
the laboratory of Nabunturan National
Comprehensive High School for at least
2 days. After the drying process, the
mass of the leaves was recorded again.
The leaves were homogenized using a
blender in preparation for the next
process.
Ethanol Leaves Extract Preparation
The homogenized Eleanor
Roosevelt croton plant were soak in the
ethanol for at least 48 hour with the ratio
of 1:1 (1 ml of ethanol and 1 gram of
plant extract). After the soaking process,
the extract was filtered using filter paper
no.1 in order to remove the solid
residue. The filtered solution was
evaporated using Modified Evaporation
Method.
Phase II. Experimentation
Collection of Blood Samples
The blood plasma was collected
from the 2 groups of both men and
women with age ranging from 20-30, 31-
40, 41-50 years old human blood was Table 1:
sampled into a 4 ml test tube containing APTT Test applied in 26 years old male
3.2% sodium citrate. The blood were and 24 years old female.
mixed and then centrifuged for 15 min.
Obtained plasma were analyze on Numbe
Mal Femal
Mean
r of Differenc
coagulation analyzer at 37°C (Kemal, Trials
e e
e
2015).
Trial 1 X X X
Testing the Extract 100%
The extracts were tested using Pure
Trial 2 X X X
Prothrombin Time (PT) test. Three trials extract
were conducted and each trial must Trial 3 X X X
have 2 dilutions (100% and control) .The Negativ
37 39
optimal volume ratio of anticoagulant to e Trial 4 38 sec.
sec. sec.
blood was 1:9. This ratio were control
maintained when obtaining blood Legend:
samples for Prothrombin Time (PT) test X=no coagulation formed
and Activated Partial Thromboplastin
Time (APTT) tests. The activated partial Table 1. Shows the result of
thromboplastin time test (APTT) was clotting time factor of male (37 sec.) with
used in determining the coagulation rate the age of 26 years old which was lesser
of blood in 3 male and 3 female blood than the clotting time of female (39 sec.)
donors with 3 different age bracket with the age of 25 years old.
measured in minutes. Table 1, 2 and 3
shows the effect of 100% concentration Table 2:
of San Francisco leaves extract and APTT Test applied in 32 years old male
control to the coagulation rate /clotting and 38 years old female.
time of blood.
Numbe
Femal Mean
Phase III. Data Collection and r of Male
e Difference
Trials
Analysis
The data of the study were Trial 1 X X X
based on the time (in minutes) of the 100%
application of 100% of Eleanor Pure Trial 2 X X X
extract
Roosevelt leaves extract and the control Trial 3 X X X
(no treatment) on the blood of the 6
donor’s with different sex and age Negativ
Trial 4 36.2 32.6 34.4 sec.
bracket. e control sec. sec.

Statistical Analysis Legend:

The researcher created a table to
show the result of male and female with
3 different age bracket with the use of
One-Way ANOVA.
X=no coagulation formed
Table 2. Shows the result of
clotting time factor of male (36.2 sec.)
with the age of 32 years old which is
greater than the clotting time of female
(32.6 sec.) with the age of 38 years old.
 Table 3: Table 4:
APTT Test applied in 42 years old male ANOVA of 100% ratio of the San
and 45 years old female. Francisco extract in different age
bracket of both male and female blood
Numbe
samples.
Femal Mean
r of Male
e Difference sum of df Mean F Sig.
Trials squares square
Between .109 2 .054 .000 1.000
Trial 1 X X X Groups
MALE Within 2674.460 6 445.743
100% groups
Pure Trial 2 X X X Total 2674.569 8
extract Between 6.907 2 3.453 .008 .992
Trial 3 X X X FEMALE Groups
Within 2605.813 6 434.302
Negativ groups
e Trial 4 36.5 36.4 36.45 sec. Total 2612.730 8
control sec. sec.
α = 0.01
Legend: Presented in table 4 are the
X=no coagulation formed critical F value for the df (2,6) in male
was 5.14 and the computed F value was
Table 3. Shows the result of .000, while the critical F value for the df
clotting time factor of male (36.5 sec.) (2,6) in female was 5.14 and the
with the age of 42 years old which was compound F value was .008. Since the
greater than the clotting time of female computed F value of male was lesser
(36.4 sec.) with the age of 45 years old. than its significance level 1.00 and the
computed F value of female was lesser
Presented in Table 1, 2 and 3 are than its significance level .992. The
the Activated Partial Thromboplastin significance level of both male and
Time Test result which shows the female were greater than the alpha level
coagulation rate (in minutes) of human (α=0.01), therefore there is no significant
blood treated with 100% concentration difference in the mean coagulation rate
of Eleanor Roosevelt croton (Codiaeum of blood in different age bracket of male
variegatum) leaves extract and the and female blood samples.
negative control (no treatment). In table
of 1, male clotting time was lower DISCUSSION
compared to female clotting time treated Based in the result on Table 1 2
with negative control, unlike in table 2 and 3, the researchers observed that the
and 3, male clotting time was higher APTT testing showed no coagulation
than the female clotting time treated with formed in Trial 1, 2 and 3 containing
the same concentration (negative Eleanor Roosevelt croton leaves extract
control). But when Eleanor Roosevelt except to the trial 4 containing negative
Croton (Codiaeum variegatum) leaves control (no treatment) which shows
extract was applied, clotting time of male different clotting time results in test tube
and female in table 1, 2 and 3 shows no containing blood plasma. After the
coagulation formed in blood samples. observation, analyzing and computing
the data result has been done. In the
result presented in Table 4, there is no
significant difference between the male
and female significant difference with
the alpha level (α=0.01). Blood samples
was taken from the 3 male and 3 female
blood donors with the age ranging from
20-30, 31-40 and 41-50. The test tube
were prepared for the experimentation.
The test tube with blood sample is
centrifuged to separate the blood
plasma to the red blood cell. The
collected blood plasma is placed in the
test tube. The plant extract is placed to
the test tube for the preparation for the
next process. The medical technologies
prepared a 4 test tube for the 3 trial and
for the control. The amount of blood
plasma and the plant extract is the
same. The researchers observed that
the test tubes with plant extract doesn’t
coagulate than the control (no
treatment). The blood samples that
treated with 100% extract concentration
showed the highest percent of delaying
the clotting time of the blood plasma
than the control using pure water.
The result of experimentation
showed that the clotting time factor of
male and female blood plasma has no
difference and this statement was
supported by the study of Saadi (2015)
that there was no significant difference
between male and female sex using
(p>0.01). Flavanoids is commonly used
compound for blood anticoagulant
because it inhibits the coagulation of
blood plasma. The study shows that the
flavanoids has a natural ability to affect
the distinct functions of platelets.
(Vallance et. al, 2019). Based on the
study of Flaumenhaft, the Eleanor
Roosevelt croton leaves extract is truly
effective because of the compound like
alkaloids and flavanoids that can inhibit
platelet aggregation or it can coagulate
the blood, also in the study of A.indica
showed that the increase of its extract
can strongly inhibits the coagulation
process and increases prothrombin
time. This may be attributed to the
presence of phytochemical compound
that have been noted in the leaves
extracts Flavonoids are proven to be the
most potently anti-thrombotic compound
that we can use. Meanwhile, alkaloids
have anticoagulant and anti-platelet
activities. This compound was found to
produce the strongest inhibition of the
platelet aggregation (Ain et al., 2016).
The delayed on blood coagulation
rate can be ascribed to the presence of
flavonoids and alkaloids on Eleanor
Roosevelt croton (Codiaeum
variegatum) leaves extract based on
phytochemical analysis. Croton,
(Codiaeum variegatum), is a common
houseplant grown for its striking foliage.
It is one of six species of broadleaf
evergreen perennials, shrubs, and small
trees in this genus in the euphorbia
family (Euphorbiaceae) which are native
to tropical Asia and the western Pacific
region (not to be confused with Croton,
another genus of more than 700 species
in the same family, in which it was
formerly incorrectly classified as Croton
(Codiaeum variegatum). In subtropical
and tropical climates they are frequently
used as landscape shrubs for dramatic
hedges, bold focal points in gardens, or
potted specimens around buildings.
(Susan, 2019).
Coagulation also called as
clotting is a process that changes the
blood from liquid to a viscous gel that
makes the blood to slowly clot. The
stopping of blood loss from a destroy
vessel, followed by restoration is called
hemostasis. Problems and diseases
takes place when the coagulation of
blood happened inside our body. If the
coagulation of blood would be detected
early, it can be treated in order to avoid
serious damaged and problems that can
cause stroke and even death. (American
Academy of Family Physicians, 2020).
Taking commercial drugs like
warfarin has side effects to our body.
Using warfarin drug gives affectivity
when used as blood anticoagulant but
other than its affectivity, it also has
complications such as dizziness,
vomiting, headache and death of skin
tissue also known as skin necrosis when
taken improperly. The main risk is
bleeding. But even though the major
bleeding is low, still, possible
complications must be notice.
CONCLUSION
Based on the results, the
researchers found out that the Eleanor
Roosevelt (Codiaeum variegatum)
leaves extract can delayed the
coagulation time rate of the blood
plasma. Therefore, the Eleanor
Roosevelt (Codiaeum variegatum)
leaves extract can be a potential source
for blood anticoagulant. This is because
of the presence of the compound
namely alkaloids and flavonoids.
RECOMMENDATION
The researchers recommend
exploring more about the Eleanor
Roosevelt croton (Codiaeum
variegatum) plant especially its
possibilities to become an antiseptic.
The researchers would also like to
recommend for this plant to make a
capsule out of its extract to have a
medicine that is more effective and more
useful to everyone.
ACKNOWLEDGEMENT
The researchers would like to
express their heartfelt gratitude to God
Almighty for giving them the strength to
make this study successful, and also to
Mrs. Leah R. Guirigay for sharing her
knowledge and guiding the researchers
to improve their study. The researchers
would like to thank Mrs. Candelaria
Bolonos for her effort and time during
their laboratory experiments. They
would like to thank Mr. Kade Aquiatan,
Mr. Tito Quiring, Mr. Jose Joemar Nono,
Ms. Laudermae Pido, Mrs. Jasmine
Nono, and Mrs. Mimi Evelia Urot for
donating their blood that the researchers
use for their research. And lastly, to their
parents who continue to support them in
financial needs of their project. To all
who help to make the study success, the
researchers would like to give their
greatest gratitude. To God be the Glory!
REFERENCES
Ashwini, U., & Asha, S. (2017). A
Preliminary Novel Findings on In
vitro Anti-coagulant Activity of
Acalypha indica linn leaf extract.
International Journal of
Pharmaceutical and Clinical
Research, 9(5), pg. 425-429.
http://impactfactor.org
Ain, Q., Khan, H., Mubarak, S. and et al
(2016). Plant alkaloids as
antiplatelet agent:drugs of the
future in the light of recent
developments.
https://www.ncbi.nlm.nih.gov/pmc
/articles/PMC5032615/
Azwanida, N. (2015). A Review on the
Extraction Methods use in
Medicinal plants, principle,
strength and limitation. Medicinal
& Aromatic plants, 3(4), pg. 1-6.
https://www.longdom.org
Duric, K., Besovic, E., Niksic, H., clots/bloodclottreatment/directoral
Muratovic, S. & Sofic, E. (2015) anticoagulants/
Anticoagulant Activity of some Wang, Z., Dou, M., Du, X., Ma, L., Sun,
Artemisia dracunculus leaf P., Cao, H. & Li, C. (2017).
extract .Bosnian Journal of Basic Influences of ABO blood group,
Medical Sciences, 15(2) pg.9-15. age, and gender on Plasma
https://www.ncbi.nlm.nih.gov Coagulation factor VII,
Junia, A. (2019). Heart failure a growing Fibrinogen, von Willebrand
threat to Filipinos. Popco factor and ADAMTS13 ss
Philippines. Weaving , G., Batstone,G.,& Jones, R.
http://business.inquirernet/20875 (2016).Age and Sex Variation in
9/ Serum Albumin Concentration:
Lee, J., Lee, W., Kim, M. et al (2017) An Observation Study . Annals of
Inhibition of platelet aggregation Clinical Biochemistry, 53(1), pg.
and thrombosis by indole 106-111.
alkaloids isolated from the edible https://www.ncbi.nlm.nih. Gov
insect protaetia brevitarsis World Health Organiaztion (2017). Sex
seulensis. Specific Association of Oral
https://www.ncbi.nlm.nih.gov/pub Anticoagulant use and
med/27997749 Cardiovascular outcomes in
Mahr, S. (2019). Croton, Codiaeum patients with Atrial Fribriliation.
variegatum. University of Journal of the American Heart
Wincosin Mandison. Association6(8).
https://wimastergardener.org/file icoagulants/icoagulants/nlm.nih.g
s/209/03/croton.pdf ov
Ramya, D., Barathi, A.,
Thirunavukkarasu P., & Asha, S.
(2017). In vitro Anticoagulant
activity of Nelumbo nucifera leaf
extract on normal healthy Blood
Plasma International Journal of
Green Medicine, 11(3).
https://www.researchgate.net
Shantha, G., Mentias, A., Inamudi, C.,
Kumar, A., Bhise, V., Patel, N.
https://www.cdc.gov/ncbddd/dvt/d
ata.html
Susan, M. (2019). Croton, Codiaeum
variegatum. A Horiculture
Information article.
https://wimaster gardener.org
Tu’akoi, K. (2015). Anticoagulants and
Antiplatelet agents
https://www.stoptheclot.org/about
 APPENDICES

Collecting of San Air drying of San Filtration of Rotary evaporation

Francisco leaves Francisco leaves at soaked San of the filtered San
at Purok-3, Linda NNCHS laboratory Francisco leaves Francisco leaves for
Nabunturan, for 3 days pure extract
Davao de Oro
 Collection of Collected blood.
blood from a
donor.

Homogenizing of Measuring the

Air dried San amount of Ethanol
Francisco leaves used for soaking

Preparation of the Blood was

test tubes for the centrifuged to
experimentations get bloodSan
Soaked plasma Soaking of San
Francisco leaves Francisco leaves
in preparation for on Ethanol
filtration

Collecting of Collecting of
blood
Measuringplasma
the plantSan
Pure extract
experimentations
amount of pure from the reagent
Francisco
San Francisco Leaves bottle
extract
leaves extract