ANTI-THROMBOLYTIC EFFECT OF Tridax procumbens AND Cosmos

caudatus EXTRACTS AS INFERRED FROM HUMAN BLOOD

COAGULATION TIME

J.F. MARIANO, A.R. MENDOZA AND A.J. SAMANIEGO

CATEGORY: LIFE SCIENCE
SUBCATEGORY: BOTANY
REGION, CITY/MUNICIPALITY: CALABARZON, CAVITE CITY
CORRESPONDING AUTHOR’S EMAIL: jfm467@ymail.com

ABSTRACT

Blood loss is one of the common problems encountered during accidents, and the time it takes to cease the
blood flow, takes so much time causing life to be at risk. This study was conducted to determine the response of
platelets to pure extracts of Tridax procumbens and Cosmos caudatus on the basis of human blood coagulation.
Disinfected fingertip was punctured with a sterile lancet. The blood was plunged into a glass slide and then mixed
with the extracts. After which, the clock was started. For every 25 seconds, one end of a toothpick is passed to the
blood drop. Once filamentous materials are seen adhering on the tip of the toothpick, the clock was stopped.

The highest mean coagulation time was observed with the untreated blood sample. On the other hand, T.
procumbens showed higher mean coagulation time compared to C. caudatus. One-Way Analysis of Variance
(ANOVA), at 5% level of significance, revealed that there was a significant difference in the mean coagulation time
of the treatments. Tukey’s pairwise comparison showed that untreated blood has significantly longer coagulation
time compared to blood treated with asteraceous plant extracts. However, blood treated with T. procumbens and C.
caudatus did not exhibit any significant difference in blood thickening time. Thus, the extracts of the two asteraceous
plants promoted the formation of fibrin strands faster than untreated blood strengthening the platelet plug during
blood clotting.

Results of the study validated that the two asteraceous plants can be powerful source of precursor of blood
factors necessary for coagulation.

INTRODUCTION Tridax procumbens (Linn.) is a spreading

annual herb grows up to 20 cm in height. This can be
Blood loss is one of the common problems found in fields, meadows, croplands, disturbed areas,
encountered during accidents. In order for them to lawns, and roadsides of tropical or semi-tropical
stop the bleeding, they use first aid treatments such as areas and are well known for their medicinal
applying direct pressure, cold applications and properties among local natives. T. procumbens is
tourniquet. But using those methods are very reputed for its wound healing activities. This plant is
complicated and may lead to more serious damage if also traditionally known for its insecticidal and anti-
not properly handled. Alternative medication can be inflammatory activities. In some tribal areas, the leaf
used to prevent blood loss caused by wounds, and juice is used to cure fresh wounds, stop bleeding, as a
this is by using plant extracts. Plants have great hair tonic (Suseela et al., 2002). In Nigeria, T.
potential of managing and treating wounds due to the procumbens is traditionally used in the treatment of
presence of wound healing properties such as typhoid fever, cough, asthma, epilepsy, diarrhea and
flavonoids, terpenoids and other phytochemical hypertension.
constituents (Thakur, et al., 2011). Those properties
are evident in the plants like asters, which are Cosmos caudatus Kunth is considered as a
traditionally used by the Native Americans to treat medicinal herb which is believed to possess the
arrow wounds (Nuffler, 2008). ability to cleanse the blood and strengthen bones due
to its high calcium content. It contains 0.3% of
proteins, 0.4% of fats and carbohydrates, it also rich

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in calcium and vitamin A. Its leaf has high the testing is specified. Objectives of the study were
antioxidant property, each 100 grams of the fresh also discussed together with the safety and
leaves have the same antioxidant property to the 2400 confidentiality of the testing procedures. From the
mg of ascorbic L-acid (Atie, 2011). participants who voluntarily joined, 30 were
randomly selected as the blood donor.
With existing information on the ability of
plants to be used as a remedy for blood coagulation, Extract Application. Disinfected fingertip
very few literature cites and compares the different was punctured with a sterile lancet. The blood was
weeds and plants of the Philippines as a potential plunged into a glass slide and then mixed with the
alternative. extracts. After which, the clock was started. For
every 25 seconds, one end of a toothpick is passed to
The general purpose of this study is to the blood drop. Once filamentous materials (fibrins)
determine the blood coagulation time in different are seen adhering on the tip of the toothpick, the
plant extracts from Family Asteraceae such as T. clock was stopped and data was recorded. The above
procumbens and C. caudatus. Specifically, it seeks to procedure was done to the 30 randomly selected
find answers to the following question: participants, taking three blood samples, which is as
big as pinhead, each.
 Do T. procumbens and C. caudatus have the
ability to hasten the formation of fibrin in blood? Statistical Analyses. The data was entered in
the computer program PAST ver. 1.42 (Hammer, et
 Is there significant difference on the blood al.,2001). Analyses of Variance (ANOVA) were
coagulation time in these plant extracts? conducted for all measured quantitative parameters.

Through this study, members of the

household can have an easy access to plants that can
be used as a remedy to minor accidents such as
abrasions and cuts. This can also serve as a
benchmark in developing medicines, utilizing the A B
extracts of asters that can be used as first aid for
blood loss.

In the field of taxonomy, this information on

the ability of the surveyed plants can be used to
effectively identify the species of Family Asteraceae
that are economically important. C D

The study was conducted from June-

August 2012 at Cavite National High School, Cavite
City.

METHODOLOGY E F

Preparation of the Materials. T.

procumbens and C. caudatus were collected from
open fields in Cavite City, washed and dried
overnight. After drying the plants, 400 grams of it
were weighed and cut into strips. The cut dried leaves
were pounded using a mortar and pestle. To get the G H
pure extract, the pounded plant leaves were filtered
by squeezing it in a clean cloth. 25 mL of the Figure 1. Procedure Used in Determining the Platelet
produced extracts were used for the experiment. Response of Blood to Aster Plants on the Basis of
Coagulation Time (A-B) Collection of Plant
Permits and Waivers. Permits and waivers Specimen; (C-D) Preparation of Plant Extracts; (E-
were given to students of IV- Alexandrite, Cavite F) Collection of Blood Sample; (G-H) Application of
National High School, School Year 2012- 2013. Aster Extracts and Documentation of Coagulation
Willingness of the students to be a donor of blood for Time

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RESULTS AND DISCUSSION Comparing the average blood coagulation
time of the three treatments, blood sample treated
Extracts from two asteraceous plants (Tridax with C. caudatus showed faster reaction, followed by
procumbens and Cosmos caudatus) were applied to T. procumbens treated blood and lastly by the
blood drops from 30 samples. The time of appearance untreated blood sample (Figure 3). This suggests that
of the fibrin strands, indication that blood coagulation there is an increased platelet response-time in the
happened, was recorded for every treatment. Results blood samples treated with the aster extracts
are summarized in Table 1. compared to untreated one.

Table 1. Human Blood Coagulation time in The extracts of the two asteraceous plants
Different Treatments promoted the formation of fibrin strands faster than
untreated blood strengthening the platelet plug during
Coagulation Time (min) blood clotting.
Treatment
Min Max Mean

T. procumbens 2.15 4.55 2.94 ± 0.53 a

Coagulation time (s)

C. caudatus 1.97 4.08 2.63 ± 0.55 ab
Control 2.67 4.33 3.73 ± 0.39 b

*mean ± standard deviation computed from 30

replicates; mean followed by the same letter are not
significant at Tukey’s pairwise comparison

Based on the results of the experiment,

coagulation time in blood samples treated with aster
extracts ranged from 1.97 to 4.55 minutes. On the
other hand, untreated blood sample formed fibrin
strands between 2.67 and 4.33 minutes, which have a
shorter range compared to the treated blood samples.
The fastest time of blood coagulation was noted on Treatment
the blood sample treated with C. caudatus while the Figure 2. Average Coagulation Time of Blood,
slowest time was noted on the blood sample applied Treated and Untreated, from 30 Samples
with T. procumbens (Figure 2).
Coagulation time (s)

Sample

Figure 1. Human Blood Coagulation Time of Samples Treated with T. procumbens and C. caudatus

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 Data were subjected to one way analysis of reaction of the juice to the blood exerts inflammatory
variance (ANOVA) and Tukey’s pairwise reaction (Yaduvanshi et al., 2011).
comparisons at 5% level of significance. The null
hypothesis subjected is that the blood coagulation CONCLUSION
times of the three treatments are not significantly
different. Results of the statistical analysis are From the data gathered from the study, it
summarized in Table 2. was concluded that species of Family Asteraceae is a
potential pro-coagulant. Cosmos caudatus and Tridax
Table 2. Summary of the Analyses of Variance at 5 procumbens showed significant effect on the blood
% Level of Significance coagulation time over the control group, therefore it
proves that the two extracts hastened the coagulation
time of blood. C. caudatus showed faster effect on
Sources of Sum of Mean F
dF the blood coagulation than T. procumbens, but as the
Variation Square Square value
data shows, there is no significant difference between
their mean coagulation. Therefore, they are both
Between groups 70017.50 2 35008.70 39.86
considered a potent pro-coagulant.
Within groups 76409.60 87 878.27
Total 146427.00 89
One-Way Analysis of Variance (ANOVA),
at 5% level of significance, revealed that there was a
Since the computed F value (39.86) is higher significant difference in the mean coagulation time of
than the tabulated F value (3.10) the null hypothesis the treatments. Tukey’s pairwise comparison showed
is rejected. Thus, coagulation time of blood treated that untreated blood has significantly longer
with T. procumbens, C. caudatus and control (no coagulation time compared to blood treated with
treatment) are significantly different at 5% level of asteraceous plant extracts. However, blood treated
significance. Tukey’s pairwise comparison further with T. procumbens and C. caudatus did not exhibit
showed that the coagulation time of the control group any significant difference in blood thickening time.
is significantly different from treatments with T. Thus, the extracts of the two asteraceous plants
procumbens and C. caudatus but no significant promoted the formation of fibrin strands faster than
difference was seen among the mean coagulation untreated blood strengthening the platelet plug during
time of T. procumbens and C. caudatus. blood clotting.

From the results of the study, it shows that Results of the study validated that the two
C. caudatus has the greatest ability of being a potent asteraceous plants can be powerful source of
pro-coagulant because of its high calcium content precursor of blood factors necessary for coagulation.
which is required in the coagulation factors. Calcium Based from human blood coagulation time
works together with vitamin K and a protein called documented, the two aster plants revealed the
fibrinogen in the clotting cascade (Parr, 2011). Just presence of anti-thrombolytic activity, however, clot
like C. caudatus, T. procumbens is also a potent pro- formation property and active components are yet to
coagulant because of its different wound healing be established out.
activities reported (Suseela et al., 2002).
Since it was proven that C. caudatus and T.
Faster formation of fibrin strands, procumbens are a potential pro-coagulant, the
strengthening the platelet plug during blood clotting, researchers would like to offer the following
in C. caudatus- treated blood can be attributed to the recommendations:
antioxidative compounds present in the plant extracts
(Huda-Faujan et al., 2009). Antioxidants are 1. determine through chemical analysis the
postulated to help control wound oxidative stress and composition and properties present in the
thereby accelerate wound healing, including blood extracts of C. caudatus and T. procumbens
clotting (Fitzmaurice, 2011). which contributed to the coagulation of
blood;
On the other hand, T. procumbens juice was
proven to have a direct stimulatory effect in the initial 2. investigate the coagulation process through
stage of blood release in wound healing due to an microscopic analysis; and
increased oxidation activity (Udupa et al., 1991).
However, blood clotting with this plant extract may 3. know other species of Family Asteraceae
be inhibited in the soon period since increased that can be also a potent pro-coagulant.

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