abstract
TRANSCRIPT
ABSTRACT
Antigonon leptopus flower extract was tested for its anticoagulant effect on
human blood. Five hundred (500.00) grams of A.leptopus flowers were macerated using a
blender and the juice were squeezed to obtain the extract. Control and experimental set-
ups with three replications each were prepared. Each replication in the control set-up was
prepared by mixing 2.00 mL of distilled water with 2.00 mL of fresh blood in a vacuum
tube while each replication in the experimental set-up was prepared by mixing 2.00 mL
of the prepared extract with 2.00 mL of fresh blood in a vacuum tube. Microscopic
observation and direct vacuum tube observations were done to note the time of blood to
visibly coagulate in each trial in each set-up. In the microscopic observation the
experimental and control set-ups had an average time elapsed for the presence of blood
coagulation of 90.67 and 26.67 minutes respectively. The experimental and control set-
ups had an average time elapsed for blood clots to be visible of 38.34 and 0.83 hours
respectively in the vacuum tube observation. Two-way ANOVA at 0.01 level of
significance, degrees of freedom: (2,5) for treatment showed that there is a significant
difference between the set-ups in both microscopic and vacuum tube observation in the
time elapsed for blood coagulation to be visible. Based on the results, the A.leptopus
flower extract can be used as an effective anticoagulant on human blood.
CADENA DE AMOR(Antigonon leptopus) FLOWER EXTRACT AS
BLOOD ANTICOAGULANT
Science Investigatory Project presented in partial fulfillment of the requirements for
graduation of the Revised Basic Education Curriculum (RBEC) to the panelists and
concerned staff of Bantayan Science High School, Bantayan, Cebu. Prepared under the
direction and guidance of Mr. Ulysis Tisado.
INTRODUCTION
Nature, Importance and Rationale of the Study
The shelf-life of human blood is just a short period of time due to the rapid
coagulation of the said body fluid. Certain medical purposes need a longer period of time
for the usage of blood (Cabatit, 1985). So, it is necessary to lengthen the shelf-life of
blood by delaying its coagulation or in other words applying a blood anticoagulant for
longer storage and time of usefulness. Anticoagulant decreases the normal range of
blood’s coagulation time; hence, blood can be stored at a longer time maintaining its
physical state and fluidity. Because of this, a study, which concerns about the possibility
of delaying the coagulation time of blood by making blood anticoagulants from plant
parts specifically A. leptopus flowers, was conducted.
A.leptopus locally known as Cadena de Amor is a climbing, perennial vine. Its
stems attain a length of 10 meters and leaves are alternate up to 10 cm long. Flowers are
white or pale to deep pink up to 2 cm long, with a 5-parted and persisted perianth. Fruit is
an ovoid achene and is about 1 cm long and broad at the base, narrowing towards the tip,
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and is loosely surrounded by the persistent lobes of the flower.
Problem and Objectives of the Study
The study aimed to answer the following problems:
1. What is the effect of A.leptopus flower extract to the coagulation time of blood as
compared to that of the distilled water?
2. Is A.leptopus flower extract a possible blood anticoagulant?
The study aimed to test if A.leptopus flower extract has a blood anticoagulant
effect. It target delaying the time of blood coagulation and lengthening the shelf-life of
blood samples. It focused on evaluating the effect of A.leptopus flower extract on the
coagulation time of blood as compared to that of the distilled water.
Limitations of the Study
The study was limited only to the extraction of A. leptopus flower, the possibility
of the said extract as blood anticoagulant and the quantitative comparison of coagulation
time of blood under both microscopic and vacuum-tube observation of the extract and
distilled water.
Chemical tests and analysis of the extract was not conducted. A two-way
ANOVA was employed for the time elapsed for blood to coagulate to be visible under
both vacuum-tube and microscopic observations.
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REVIEW OF RELATED LITERATURE
Coagulation
Coagulation is a complex process by which blood forms clots. It involves a
cellular (platelet) and proteins (coagulation factors) (http://medical-
dictionary.thefreedictionary.com/Coagulation+Disorders). Blood coagulation is the
process of transformation of a liquid into a soft, semisolid, or a solid mass. Blood
coagulation is characterized by thrombosis-the formation, presence, or development of a
thrombus, a blood clot formed (Cabatit, 1985).
Coagulation Time
The coagulation time is the time by which the blood takes a clot. Normally in an
ordinary skin puncture, blood coagulation takes place in 2-6 minutes. It is affected by
such factors as temperature, size of the drop, and smoothness and cleanliness of the
instrument. But the normal coagulation time when blood is taken from a vein may reach
twenty minutes. (Cabatit, 1985)
Cadena de Amor
Chain of love, Antigonon leptopus, locally known as Cadena de Amor is a
climbing, perennial vine. Its stems attain a length of 10 meters and leaves are alternate up
to 10 cm long. Flowers are white or pale to deep pink up to 2 cm long, with a 5-parted
and persisted perianth. Fruit is an ovoid achene and is about 1 cm long and broad at the
base, narrowing towards the tip, and is loosely surrounded by the persistent lobes of the
flower. Studies have shown that it has anti-thrombin, analgesic, anti-inflammatory, ant-
diabetic and lipid peroxidation inhibitory properties. The anti-thrombin property was
observed under a methanol extract of the plant.
A.leptopus is found to be edible and practiced by some as medical remedies for
certain ailments and diseases. In some parts of the world, the tubers and flowers are
consumed as food; on the other hand, in Thailand, leaves are fried, coated with flour and
served with noodles. Ifugao in the Philippines, use Cadena de amor for wound closure. In
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Trinidad and Tobago, the plant is used for diabetes, low blood pressure, and as a heart
tonic. In Jamaica, decoction of flowers and aerial parts are used for cold remedies.
(http://stuartxchange.com/CadenaDeAmor.html)
Coagulation Components
There are many factors and substances that make up the coagulation process
namely platelets, platelet aggregators, and fibrin-formed from fibrinogen. The
coagulation system is comprised by coagulation factors: Factor XII, Factor XI, Factor X,
Factor VII and Factor V.
On the other hand, coagulation is prevented and delayed by application of blood
anticoagulants, vasoconstrictors, and platelet aggregator inhibitors. Blood anticoagulants
include heparin, coumadin and many more.
(http://www.naturdoctor.com/Chapters/Research/thrombosis_prevention.html)
Coagulation as a Complex Process
The sequence of events involving the process of coagulation (after tissue injury
and platelet destruction) are:
1. Platelet factors + Plasma factors →→→→→→→ Plasma thromboplastin
(AHG, PTC, PTA)
2. Plasma thromboplastin + (LF, SF, Ca) →→→→→→→ thromboplastin complex
3. Thromboplastin complex + Prothrombin →→→→→→→ thrombin
Platetelet acc. 1
4. Thrombin + fibrinogen →→→→→→→ Fibrin (insoluble clot)
Platelet acc.2
In the conversion of fibrinogen to fibrin, the 3 steps are:
1. proteolysis of fibrinogen
2. soft clot formation
3. hard clot formation
The clot formation is affected by the pH because the fibrin monomer aggregation of the
soft clot formation is affected by the strength of the hydrogen bonding and pH.
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(Cabatit, 1985)
Blood anticoagulants
Here are some inhibitors of blood coagulation or anticoagulants:
1. Heparin- a complex carbohydrate and is believed to be formed by the mast cells
and rapidly destroyed by heparinase. It inhibits activatiuon of prothrombin and
inactivates thrombin.
2. Antithrombin- present in small amount in plasma and inactivates thrombin.
3. Antithromboplastin- a lipid that inactivates thromboplastin.
4. Dicumarol- prevents the body from utilizing vitamin K: thus, it depresses the
production of proconvertin which is needed for blood coagulation.
5. Fibrinolytic substances- present in blood and tissues of normal persons. It limits
intravascular clotting.
Water as Amphoteric
At pH 7 water is neutral.But...water is amphoteric- it can act as an acid or a base
if the pH of 7 is disrupted. Its acidic or base like properties depend on whether it is
receiving or donating a proton. (acids-proton donors, bases-proton acceptors).
(http://wiki.answers.com/Q/Is_water_a_base_or_an_acid)
Statistical Parameter
A two-way Analysis of Variance is a statistical parameter to determine if there is
a significant difference between the existing treatments and replications. It is used if 2 or
more variables are involved.
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METHODOLOGY
Materials:
Blender
pH paper
wire gauze
125.00 mL Erlenmeyer flask
106.00 mL Distilled water
Dropper
12.00 mL Fresh blood samples
12 vaccuum tubes
3 microscopes
slides and cover slips
250.00 mL beaker
alcohol lamp
iron rings
iron stands
100.00 mL graduated cylinder
Cheese cloth
Triple beam balance
500.00 g A.leptopus flowers
GENERAL PROCEDURE:
*Collection of flowers
Pink-colored flowers of A.leptopus were collected and weighed. Before weighing
the fruits were separated from the flowers.
*Extraction A. leptopus flowers
Five hundred (500.00) grams of collected A.leptopus flowers were macerated with
100.00 mL of distilled water using a blender. The macerated flowers were decanted to a
cheese cloth and were squeezed to obtain the extract. The extract was transferred into an
e-flask. The pH, volume, color and odor of the extract were noted.
*Cleaning of vacuum tubes
The vacuum tubes were sterilized by subjecting them to boiling water placed in a
beaker.
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*Preparation of the Reagents
Three vacuum tubes were filled with 2 mL each of the prepared extract. This
composed the experimental set-up. Another three vacuum tubes were filled with 2 mL
each of distilled water, which composed the control set-up.
*Collection of Blood Samples
Blood was freshly extracted from two persons, labeled as A and B, by the Medical
Technician of Bantayan District Hospital. Six vacuum tubes were filled by 2 mL each of
the freshly extracted blood.
*Preparation of the Set-ups
Right after the blood was decanted into the vacuum tubes, they were immediately
mixed with the prepared reagents namely the A. leptopus flower extract and the distilled
water. The following set-ups were made:
Set-ups Trial 1 Trial 2 Trial 3
Experimental 2 mL blood(A) + 2
mL A. leptopus
flower extract
2 mL blood(B) + 2
mL A. leptopus
flower extract
2 mL blood(A) + 2
mL A. leptopus
flower extract
Control 2 mL blood(A) + 2
mL distilled water
2 mL blood(B) + 2
mL distilled water
2 mL blood(B) + 2
mL distilled water
*A and B are the source of the blood
*Microscopic Observation
The set-ups were continuously observed under the microscope (400x) by getting a
small amount of each sample using a dropper and putting them into the slides. After the
slide was used it was subjected to boiling water for sterilization for future use.
Supposedly, the observation interval must be one minute, but, considering that getting the
specimens, putting them into the slides, and focusing them clearly under the microscope
takes more than two minutes, a fixed interval was already neglected. Instead of fixing the
interval, an immediate continuous microscopic observation was made. It means that after
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a few minutes of observing under the microscope for determining the presence of blood
coagulation, new samples were immediately obtained and subjected to the said
observation. One researcher was responsible for taking the notes of every microscopic
observation finished and was the one taking the exact time for the said confirmation of
the presence of blood coagulation as viewed microscopically for every trial in each set-
up. Each time the blood coagulation was present and observed in a certain blood sample,
further microscopic observation to that certain sample was already being stopped.
*Vacuum tube observation
Simultaneous to the microscopic observation, a vacuum-tube observation was also
conducted by viewing from the image formed as being viewed at the outside of the
vacuum-tube if it had visibly coagulated already. The time and date for insoluble blood
clot to be visible under vacuum tube observation for each of the trials of every set-up was
noted.
*Data Processing
The data obtained on both microscopic and vacuum tube observation cannot be
used as a statistical data; hence, the obtained data were processed for presentation of
results and statistical use.
The time (in minutes) elapsed for the presence of blood coagulation under
microscopic observation was obtained by using the equation below.
(HH°MM’)f - (HH°MM’)i = HH° x + MM’
where:
(HH°MM’)f - noted time for presence of blood coagulation
(HH°MM’)i - time for blood sample to be freshly obtained
HH° x + MM’ – time elapsed in minutes
HH° - hour time
MM’ - minute time
To obtain the time elapsed, we subtract the noted time for the presence of blood
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( )
60’
1°
( )
60’
1°( )
( )
( )
coagulation by the time for the blood sample to be obtained. The equation neglected the
date and time position (AM/ PM) because the data obtained had the same date, February
3, 2012, and time position, PM. The elapsed hour time was converted into minutes using
a conversion factor and then added with the elapsed minute time to obtain the time
elapsed in minutes for the presence of blood coagulation under microscopic observation.
The time (in hours) elapsed for insoluble blood clots to be visible was obtained by
using the equation below.
[ (DDf- DDt) x X + (HH°MM’)f] - (HH°MM’)i = HH° + MM’ x
where:
(HH°MM’)f - noted time for presence of blood coagulation
(HH°MM’)i - time for blood sample to be freshly obtained
HH° - hour time in 24- hour time format
MM’ - minute time
DDf - noted date for insoluble blood clot to be visible
DDt - date for blood sample to be freshly obtained
X - 12° if (HH°MM’)f is less than (HH°MM’)i
- 24° if (HH°MM’)f greater than (HH°MM’)i
[ (DDf- DDt) x X + (HH°MM’)f] – modified noted time for insoluble blood clot
to be visible
HH° + MM’ x - time elapsed in hours
To obtain the time elapsed, first, the date for blood sample to be freshly obtained
subtracted from the noted date for insoluble blood clot to be visible. And then, multiplied
the obtained value with the corresponding hour equivalent and added it to the noted time
for insoluble blood clots to be visible. The obtained value was already the modified noted
time. We subtract the modified noted time for insoluble blood clots to be visible by the
time for the blood sample to be obtained. The elapsed minute time was converted into
hours using a conversion factor and then added with the elapsed hour time to obtain the
time elapsed in hours for insoluble blood clot to be visible under vacuum tube
observation.
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1°
60’( )
( ))1°
60’
( )
*Incineration of Blood Samples
After the blood samples were already observed by all the data and information
needed, they were incinerated and the vacuum tubes were disposed for safety and sanitary
purposes.
*Statistical Parameter and Analysis
Since the statistical parameter used is two-way ANOVA, we need to define our
hypotheses for replication and treatment to interpret our results in the given parameter.
The hypotheses for replication are the following:
Null hypothesis: There is no significant difference between the time (in minutes) elapsed
for blood to coagulate under microscopic observation or time (in hours) elapsed for
insoluble blood clots to be visible under vacuum tube observation in every replication or
trial; therefore,
Ho is R1 = R2 = R3 where: R- replication or trial
Alternative hypothesis: There is a significant difference between the time (in minutes)
elapsed for blood to coagulate under microscopic observation or time (in hours) elapsed
for insoluble blood clots to be visible under vacuum tube observation in every replication
or trial; therefore,
Hi is R1 ≠ R2 ≠ R3 where: R- replication or trial
The hypotheses for treatment are the following:
Null hypothesis: There is no significant difference between the time (in minutes) elapsed
for blood to coagulate under microscopic observation or time (in hours) elapsed for
insoluble blood clots to be visible under vacuum tube observation in every treatment-
blood with A. leptopus extract and blood with distilled water; therefore,
Ho is T1 = T2 where: T- treatment
Alternative hypothesis: There is a significant difference between the time (in minutes)
elapsed for blood to coagulate under microscopic observation or time (in hours) elapsed
for insoluble blood clots to be visible under vacuum tube observation in every treatment-
blood with A. leptopus extract and blood with distilled water; therefore,
10
( )( )( )
Hi is T1 ≠ T2 where: T- treatment
If the obtained F value is greater than or equal to the tabular value at 0.01 level of
significance, degrees of freedom: (2,5) for treatment and (1,5) for replication, null
hypothesis(Ho) was rejected and alternative hypothesis(Hi) was accepted.
The results of this two-way ANOVA are favorable to the study if there is no
significant difference between the existing replications. And if there is a significant
difference between the treatments, since the comparison is between the A. leptopus
flower extract- the tested variable as blood anticoagulant- with the distilled water- an
unconsidered anticoagulant.
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RESULTS AND DISCUSSIONS
Physical Properties of the A. leptopus Flower Extract
Part of the study was the description of the A. leptopus flower extract’s physical
properties and characteristics. The macerated 500.00 g A. leptopus flowers yielded an
extract with the following physical characteristics:
Table 1.1. Physical properties of the A. leptopus flower extract
Characteristic Description
Color Pale-brown
Odor Strong, woody odor
pH 4
Volume 70 mL
The A. leptopus flower extract had a pale-brown color, strong, woody odor, and a
pH of 4. The obtained extract from the macerated flowers with 100.00 mL distilled water
was 70 mL. The extract was acidic based on its pH value. The acidic property of the
extract may possibly be caused by the organic materials and other substances contained
by the A. leptopus flowers. Those organic materials and substances were obtained by the
extract upon squeezing the macerated flowers. The low pH value or the acidic nature may
possibly affected the time of clot formation or blood coagulation as it is cited on the
Review of Related Literature (see Review; Coagulation as a Complex Process). The
extracted volume, 70 mL, was less than that of the volume of the added water, 100 mL,
may be attributed by: a. the minimal amount of water which remained in the flowers in
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the blender during decantation ( as it is observed that the A. leptopus flowers absorbed
water); b. the existing amount of water absorbed by the flowers in the residue after the
squeezing ( as it is considered that squeezing is not enough to extract the whole amount
of water that is contained by the macerated flowers).
Microscopic Observation
The presence of blood coagulation can be observed microscopically by viewing if
there is already a presence of cells that have already colonized or grouped or clumped,
and if viewed at technically high microscopes, there is already the presence of a clear
thrombus. The time by which this presence of coagulation occurs is important because it
will lead to the determination of the coagulation time of a certain blood sample. Table 2.1
below shows the elapsed time of blood for its coagulation as it was viewed
microscopically.
Table 2.1. Time (in minutes) elapsed for the presence of blood coagulation under
microscopic observation
Set-up Trial 1 Trial 2 Trial 3 Average
Experimental 98 88 86 90.67
Control 36 18 26 26.67
Table 2.1 shows the time difference of the time which the blood samples was
observed to have the presence of blood coagulation from the time by which the blood
sample was freshly obtained and was mixed by the reagents. The control set-up trial 2,
obtained the earliest time which is 18 minutes for blood coagulation to be observed,
13
followed by control set-up trial 3 which coagulated 26 minutes after it was obtained, then,
followed by control set-up trial 1 which had a time elapsed of 36 minutes. The
experimental set-up trial 3 coagulated in the 86 th minute, and then experimental set-up
trial 2 coagulated after 88 minutes and lastly experimental set-up trial 1 was observed to
have the presence of blood coagulation 98 minutes after the blood was freshly obtained.
The coagulation time of the blood in control set-up trials 1 and 3, which were 36
and 26 respectively, were more than the normal time of coagulation, which was 20
minutes, as presented in the review. This few minutes of delay might be caused by the
presence of distilled water, which may sometimes act as an acid and sometimes as a base.
The nature of the distilled water of its basicity and acidity may also have effected the
coagulation time of blood. ( see Review; Water as Amphoteric).
*Statistical Analysis
The two –way Analysis of Variance (ANOVA) is essential to determine whether
or not there is a significant difference between the tested variables. We are also able to
determine if there is a significant difference between the replications as presented on the
data utilized. This two-way ANOVA will be able to lead us to an interpretation if the A.
leptopus flower extract can be used as a blood anticoagulant. Table 2.2 shows the two-
way ANOVA of the time elapsed for the presence of blood coagulation under the
microscope.
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Table 2.2. Two-way Analysis of Variance (ANOVA) for the time (in minutes) elapsed
for the presence of blood coagulation under microscopic observation
Source of
Variance
Degrees of
freedom
Sum of
Squares
Mean
SquaresF value
Table value
(0.01)
Replication 2 217.33 108.67 15.52* 18.00
`Treatment 1 6144 6144 877.71** 21.20
Error 4 28 7 N/A N/A
**-significant *- not significant
As stated above in the table, there are three sources of variance: the replication,
treatment and error. The degrees of freedom of replication and treatment, 2 and 1
respectively, were obtained from the total number of replication of treatments decreased
by 1. The degrees of freedom of error, 4, was from the sum of the total number of
replications and treatments decreased by 1. The sum of squares for replication, treatment,
error were 217.33, 6144, and 28 respectively. The mean squares for replication,
treatment, error were 108.87, 6144, and 7 respectively obtained by dividing the sum of
squares by their respective degrees of freedom. The F value for replication and treatment,
15.52 and 877.71 respectively, were obtained by dividing each mean square by the mean
square of error. The obtained F value for replication, 15.52, is less than the critical value
which is 18.00 at 0.01 level of significance; therefore, the Ho is accepted which states
that there is no significant difference between the time (in minutes) elapsed for the
presence of blood coagulation under microscopic observation in the three trials or
replications. It can then be interpreted that the 3 trials or replications has obtained
15
acceptable data from the accepted Ho that the trials have insignificant differences with
each other; hence the three trials had a just and fair data based from the statistical result.
The F value for treatment, 877.71, is greater than the critical value, 21.20, at 0.01
level of significance. So, there is a significant difference between the the time (in
minutes) elapsed for the presence of blood coagulation under microscopic observation of
the experimental set-up, the blood with A. leptopus flower extract, and the control set-up,
the blood with distilled water. It can then be concluded that the A. leptopus flower extract
has an effect on the delay in the the time (in minutes) elapsed for the presence of blood
coagulation under microscopic observation.
Vacuum tube Observation
The naked eye observes and detects the presence of blood clot as the blood
sample is seen on a transparent container like a vacuum tube. The time of the presence
the blood clot had been observed will identify the time by which the blood sample
produces an observable, insoluble blood clot, which is a product of blood coagulation.
Table 3.1 shows the time (in hours) elapsed for insoluble blood clots to be observed as
observed from the outside of the vacuum tube.
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Table 3.1. Time (in hours) elapsed for insoluble blood clots to be visible under vacuum
tube observation
Set-up Trial 1 Trial 2 Trial 3 Average
Experimental 49.18 49.15 16.70 38.34
Control 0.85 0.82 0.82 0.83
Table 3.1 shows the time difference of the time which the blood samples was
observed to have the presence of blood coagulation from the time by which the blood
sample was freshly obtained and was mixed by the reagents. The control set-up trials 2
and 3 obtained the earliest time elapsed which is 0.82 hours for blood clots to be
observed. The remaining trial, trial 3, of the control set-up obtained the next time
difference with 0.85 hours. The trials of the experimental set-up garnered the longest time
that elapsed. Trial 3 of the said set-up had a 16.7 time elapsed in hours. Next with 49.15
hours of elapsed time, trial 2 was observed with blood clots. And lastly, trial 1 had the
longest time difference with 49.18 hours of elapsed time for insoluble blood clots to be
observed.
The very early coagulation of trial 3 of the experimental set-up as compared to the
other trials in the same set-up may possibly be caused by the following events which
occurred during the experiment:
In taking samples for the blood to be used for microscopic observation, a dropper
was just utilized. Because the instrument is not that appropriate for taking blood
specimen, spills, decantation errors and other wasteful scenarios had occurred. So, after
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the microscopic observation was already done, trial 3 was observed to have the least
volume remaining in the tube as compared with the other trials. Since the extract was less
dense than the blood (as it is observed that the extract floats over the blood in the mixing
process), it was more prone to waste and spills. Since the A. leptopus extract is one of the
independent variables, which causes the change in the dependent variable- the time for
blood clots to be visible under vacuum tube observation- it is possible that the less
volume of the extract that remained in the trial caused the early delay of blood clots to be
visible as compared to that of the other trials in the experimental set-up.
* Statistical Analysis
The two –way Analysis of Variance (ANOVA) is essential to determine whether
or not there is a significant difference between the tested variables. We are also able to
determine if there is a significant difference between the replications as presented on the
data utilized. This two-way ANOVA will be able to lead us to an interpretation if the
blood with A. leptopus extract can delay the time elapsed for blood clots to be visible.
Table 3.2 shows the two-way ANOVA of the time elapsed for the presence of blood
coagulation as it is observed in the vacuum tube.
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Table 3.2.Two-way Analysis of Variance (ANOVA) for the time (in hours) elapsed for
insoluble of blood clots to be visible under microscopic observation
Source of
Variance
Degrees of
freedom
Sum of
Squares
Mean
SquaresF value
Table value
(0.01)
Replication 2 351.65 175.83 2.00* 18.00
`Treatment 1 2110.87 2110.87 24.06** 21.20
Error 4 351 87.75 N/A N/A
**-significant *- not significant
As stated above in the table, there are three sources of variance: the replication,
treatment and error. The degrees of freedom of replication and treatment, 2 and 1
respectively, were obtained from the total number of replication of treatments decreased
by 1. The degrees of freedom of error, 4, was from the sum of the total number of
replications and treatments decreased by 1. The sums of squares for replication,
treatment, and error were 351.65, 2110.87, and 351 respectively. The mean squares for
replication, treatment, and error were 108.87, 6144, and 7 respectively obtained by
dividing the sum of squares by their respective degrees of freedom. The F value for
replication and treatment, 2.00 and 24.06 respectively, were obtained by dividing each
mean square by the mean square of error. The obtained F value for replication, 2.00, is
less than the critical value which is 18.00 at 0.01 level of significance; therefore, the Ho
is accepted which states that there is no significant difference between the time (in hours)
elapsed for blood to coagulate under vacuum tube observation in the three trials or
replications. It can
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then be interpreted that the 3 trials or replications had obtained acceptable data from the
accepted Ho that the trials have insignificant differences with each other; hence the three
trials had a just and fair data based from the statistical results.
The F value for treatment, 877.71, is greater than the critical value, 21.20, at 0.01
level of significance. So, there is a significant difference between the the time (in hours)
elapsed for blood to coagulate under vacuum tube observation of the experimental set-up,
the blood with A. leptopus flower extract, and the control set-up, the blood with distilled
water. It can then be concluded that the A. leptopus flower extract has an effect on the
delay in the time (in hours) elapsed for blood to coagulate under vacuum tube
observation..
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CONCLUSIONS AND RECOMMENDATIONS
CONCLUSIONS
Based from the interpretations of the statistical parameter on both time elapsed for
visible blood coagulation under microscopic and vacuum tube observations, there is a
delay in the coagulation perion of blood with A. leptopus flower extract as compared to
that of the distilled water. So, by delaying the coagulation time , it will lengthen the shelf-
life of blood. In summary, Cadena de Amor (Antigonon leptopus) flower extract can be
used as a blood anticoagulant.
RECOMMENDATIONS
Conduct a chemical test on the A. leptopus flower extract to know what
component of the extract causes the anticoagulant activity on human blood
Compare the effectiveness of the extract as blood anticoagulant with existing and
commonly used blood anticoagulants
Let the treated blood undergo in a centrifuge machine to clearly view the size and
presence of the insoluble blood clots
Utilize a more appropriate device or instrument for taking the specimen to be used
in microscopic observation for better accuracy and fairness to each trial
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Conduct a study if the A. leptopus flower extract has an effect to the physical and
chemical characteristics and composition of blood other than coagulation time
which may affect the needed blood components for medical and other purposes
Use a blood sample from one person only throughout the experiment for a more
just experiment
Utilize a more highly powered and technically high microscopes for easy and fast
viewing of blood coagulation; in this case, the microscopic determination would
be easier and faster; thus, it would take a little time and a fixed small time interval
can be utilized in the microscopic observation, which produces a more accurate
result
Conduct another study which focuses on the effectiveness of A. leptopus flower
extract with respect to the ratio of the volume of the extract and the volume of the
blood sample treated.
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BIBLIOGRAPHY
Cabatit (Espino, Belen). Biochemistry. Quezon City: NA, 1985
http://medical-dictionary.thefreedictionary.com/Coagulation+Disorders
http://stuartxchange.com/CadenaDeAmor.html
http://www.naturdoctor.com/Chapters/Research/thrombosis_prevention.html
http://wiki.answers.com/Q/Is_water_a_base_or_an_acid
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