Hemostatic Effects of Peperomia Pellucid
Hemostatic Effects of Peperomia Pellucid
Hemostatic Effects of Peperomia Pellucid
In Partial Fulfilment
Of the Requirement for the Degree
Bachelor of Science in Biology
Norlainie C. Omar
March 2013
Table of Contents
Title Page i
Approval Sheet ii
Acknowledgment iii
Dedication iv
Table of Contents v
Appendices vi
List of Tables vii
List of Plates ix
Abstract x
CHAPTER I: Introduction
Background of the Study 1
Statement of the Problem 2
Research Hypothesis 3
Significance of the Study 4
Objectives of the Study 4
Scope and Limitations 5
v
Duncan’s Multiple Range Tests 18
Mean Values 19
Literature Cited 39
Appendices
List of Tables
List of Figures
List of Plates
vi
APPENDICES
vii
LIST OF TABLES
viii
LIST OF PLATES
ix
ABSTRACT
Omar, N. March 2013. “Hemostatic effects of Peperomia pellucida plant crude extract, in
vitro.” Biology Department, College of Natural Sciences and Mathemetics, Mindanao State
University, Marawi City.
overwhelming consumer response seeking remedies devoid for unfavorable side effects has
prompted the execution of this study. The study was conducted to determine the coagulation
known for its traditional use. The study is helpful to the people in the community because this
Blood samples from the donors were tested for coagulation through clotting time assay.
To test the coagulation properties of the Pansit-pansitan (Peperomia pellucida) plant, four
treatments were evaluated. The first treatment serves as the control variable with 0.2ml of whole
blood. The second treatment was a mixture of 0.2ml of 50% crude extract and 0.2ml of whole
blood. The third treatment was a mixture of 0.2ml of 75% crude extract and 0.2ml of whole
blood. And the fourth treatment was a mixture of 0.2ml of 100% crude extract and 0.2ml whole
blood. Each treatment had five replicates. The resulting mixtures were scrutinized within a
certain period of time to determine the effectiveness of the crude extract for blood coagulation by
The results of the study showed that Pansit-pansitan (Peperomia pellucida) plant had
coagulation properties that can induce blood clotting and augmentation of thrombocytes
production.
x
Chapter I
INTRODUCTION
Plants have played a significant role in maintaining human health (Craig, 1999) and
improving the quality of human life for thousands of years and have served humans well as
Piperaceae (Ghani, 1998), commonly known as shiny bush (Sio, 2012) or pansit-pansitan (Cao,
2012). It is widely distributed in most South American and Asian countries (Bayma, 2008).
According to Ethno-botanical studies, the whole plant has been in medicinal use since long
treat abdominal pain, abscesses, acne, boils, colic, fatigue, gout, headache, renal disorder and
rheumatic joint pains (Khan, 2002; Ragasa, 1998; Omoloso, 2002). The roots are used to treat
fevers and the aerial parts are used as dressing for wounds (Muñoz, 2000). The whole plant is
used to stop hemorrhages (Egwuche, 2011); it is crushed and mixed with water to form a
mixture, heated and administered orally or applied topically to arrest wound bleeding or
pathological of the vessels or by traumatic rupture of one or more vessels (Wagman, 2000). The
1
blood has the ability to change from a fluid to a solid and back to a fluid again. The change to a
Hemostasis, when a small blood vessel is transected or damaged; the injury initiates a
series of events that leads to the formation of a clot (Ganong, 2000). It leads to the sealing off
blood vessel preventing further blood loss (Ganong, 2000). A wound or cut on blood vessels
causes vasoconstriction and thrombin activation which are then accompanied by adhesion and
platelet activation, fibrin formation and coagulation activation mechanism. Thrombosis is the
pathological formation of hemostatic plug within the vasculature in the absence of bleeding
(Rang, 1999).
blood clot conversion process. Medical treatment includes administration of drugs either
locally (topical) or systemically (oral or parenteral) with the aim to hasten the time required for
blood clotting. Medicinal plants have generated much interest for treatment of wound bleeding
as they are affordable and purportedly safe from hypersensitive reactions (Khan, 2010).
hemorrhage, which is cost-efficient, effective, and safe conceived the idea of doing an
interventional study to determine the effects of Peperomia pellucida plant extract to the normal
2
Statement of the Problem
Peperomia pellucida is one of the most studied plants in terms of its ethno-medicinal
values (Ytable, 2012). However, studies on the hemostatic activity of this plant are rather
limited. Despite its wide range of folk medicinal use, there is insufficient scientific data
(Majumder, 2011) on the efficacy of this plant in arresting wound bleeding or hemorrhage.
Most of the reports are anecdotal therefore, an investigation must be made to verify these claims.
The present aim of the study is to provide answer to the following questions:
1. Will Peperomia pellucida plant extract induce an effect on the normal clotting time
of human blood?
2. Will Peperomia pellucida plant extract cause to hasten the normal clotting time of
human blood?
3. Will Peperomia pellucida plant extract cause to prolong the normal clotting time of
human blood?
Research Hypotheses
Null: Induction of Peperomia pellucida plant crude extract has no effect on clotting of
3
Significance of the Study
hemorrhage will be scientifically justified. This study is also a significant attempt on combating
major morbidity and mortality rate caused by hemorrhage (Goker, 2008) by obtaining
hastening the normal blood clotting or thrombin time. Thus, a positive result of this study will
serve as an identification of a novel, effective hemostatic agent that improves the management of
bleeding in a wide range of patients from all disciplines of clinical medicine (Goker, 2008). This
will be inexpensive and readily available since the test plant is locally found and grown in this
country (Mustapha, 2012). This could be a great help in developing countries in utilizing
properly their environmental resources. Furthermore, the research work would give an idea to
the people to cultivate the commonly found plants such as Pansit-pansitan in our country with
valuable medicinal use and would serve as an avenue for further discovery in research and for
The main objective of the study is to determine whether the plant extract of Peperomia
pellucida contain hemostatic components. Specifically, the researcher aims to accomplish the
following:
1. To determine whether or not the plant extract of Peperomia pellucida induce an effect
4
2. To determine whether or not the plant extract of Peperomia pellucida will cause to
3. To determine whether or not the plant extract of Peperomia pellucida will cause to
This study is concerned in knowing the possible effects of Peperomia pellucida plant
extract on the normal clotting time of human blood. This study utilized only locally available
Peperomia pellucida plant. The entire plant was utilized for extraction by pounding, squeezing
and filtration method. The experimental subjects used were human blood drawn from donors via
The experimental design used in this study was Random Complete Block Design
(RCBD). There were three blocks with four treatments, each treatment with five replicates
(n=60). All Treatments 1 received no crude extract, all Treatments 2 received 50% crude extract,
all Treatments 3 received 75% and all Treatments 4 received 100% crude extract. The notable
effects of plant crude extract on hemostasis was determined by noting blood clotting time on
normal human blood. Appropriate units of plant extract were added immediately to normal
human plasma respectively, the time it took for blood to clot was noted in minutes. Descriptive
statistics of the data results were expressed as Mean ± Standard Deviation (S. D.) using Two-
way Analysis of Variance (ANOVA) and Duncan’s Multiple Range Test (DMRT).
Phytochemical analyses of the nutrient component of the test plant are beyond the scope
of this study. The isolation of active components that may have influence alteration of normal
5
clotting time in human blood were not determined in this study. Other basic mechanisms for
action of hemostatic activity of the plant extract are excluded in this study.
6
Chapter II
A. Experimental Plant
TAXONOMIC CLASSIFICATION
Kingdom: Plantae
Division: Magnoliophyta
Class: Magnoliopsida
Order: Piperales
Family: Piperaceae
Genus: Peperomia
Species: P. pellucida
Piperaceae (Ghani, 1998). It is commonly known as shiny bush. It is found in various shaded,
damp habitats all over Asia and America, growing in clumps, thriving in loose, humid soils,
characterized by succulent stems, shiny, heart-shaped, fleshy leaves and tiny, dot-like seeds
As ethno-medicinal uses of this plant Peperomia pellucida has been applied for treating
abdominal pain, abscesses, acne, boils, colic, fatigue, gout, headache, renal disorders and
rheumatic joint pain (Majumder, 2011). The roots are used to treat fevers and the aerial parts are
used as dressing for wounds (Muñoz, 2000). The plants have been used as a hypocholesteremic
7
agent (Bayma, 2000). It is a popular cough suppressant, emollient and diuretic as well as
effective in the treatment of proteinuria (Theresa, 2012 as cited by Blank, 2002). The decoction
of the plant is used in the Philippines to decrease uric acid levels and to treat renal problems. It
is also used topically for skin disorders such as acne and boils (Egwuche, 2011). The plant is
described to passify vitiated cough, pitta, constipation, kidney diseases, urinary retention,
dysuria, urinary tract infections, emaciation, edema and general weakness. Infusion and
decoction of leaves and stems of fresh plant are eaten as salad for the treatment of gout and
arthritis (Cao, 2012). According to ethno-botanical studies, the whole plant has been in
medicinal use since long. It is crushed and mixed with water to form a mixture, heated and
administered orally to cure hemorrhage (Egwuche, 2011; Majumder, 2011; Theresa, 2012).
Literature of many researchers prove that the plant contain alkaloids, saponins, tannins
and cardenolides (Khan, 2010) flavonoids, essential oils and carotol (Khan, 2002).
Plants have long played a significant role in maintaining human health and have served as
food for humans. WHO estimated that over 80% of the earth’s inhabitants rely on traditional
medicine for their primary health care needs, and most of this therapy involves the use of plant
Plants evolved the ability to synthesize chemical compounds that help them defend
against attack from a wide variety of predators. By chance, some of these compounds, while
being toxic to plant predators, turn out to have beneficial effects when used to treat human
diseases (Ytable, 2012). Traditional use of medicines is recognized as a way to learn about
8
potential future medicine. In 2001, researchers identified 122 compounds used in mainstream
medicine that were derived from “ethno-medical” plant sources; 80% of these compounds were
Both in modern and traditional medicine, medicinal plants continue to provide valuable
therapeutic agents in the cure of disease and aliments (Davison, 2006). Doubts about the
efficacy and safety of the oral hypoglycemic agents have prompted a search for safer and more
effective drugs in the treatment of many diseases (Davison, 2006 as cited by Bannerman, 1993).
Many herbs have remained as an alternative to conventional therapy especially in poor areas
where insulin is not readily available; due to the high cost and the lack of medical aid (Davison,
Medicinal plants are the oldest known health-care products, where renewed interest is
growing based on the ethnological, medical and historical background of each country (Davison,
2006). Medicinal plants have always been important for pharmacological research and drug
development, where the plant constituents are used directly as therapeutic agents, for the
synthesis of drugs or as models for pharmacologically active compounds (Davison, 2006 as cited
by Levetin, 1999).
The plant based on indigenous knowledge was passed down from generations in various
parts of the world throughout its history and has significantly contributed to the development of
different traditional systems of medicine. More recently, drug discovery techniques have been
(Jachak, 2007).
9
The use traditional medicine has increased in developed countries also, mainly due to the
failure of modern medicine to provide effective treatment for chronic diseases and emergence of
multi-drug resistant bacteria and parasites. The current emphasis of new drug discovery
processes from plants is the development of products with new pharmacological modes of
C. Hemorrhage
Blood loss, while minor in every day cuts and bruises, is one of the main causes of
mortality. Hemorrhage threatens the life safety of patients and the wounded in trauma care and
surgical intervention. Hemorrhage is the main reason in the causes of death in 48 h after trauma,
which accounts for 80% in all trauma accident (Wang, 2011). Early control of hemorrhage
remains the most effective strategy for treating combat casualties. Catastrophic blood loss often
and save patient’s life in the trauma is of medical importance (Wang, 2011).
Controlling hemorrhage will always remain a top priority in trauma care, and the
development of materials to achieve this goal more effectively is of obvious benefit. In response
to the changing combat and trauma casualty care, there has been an increase in efforts to develop
better hemostatic agents. An ideal agent should be effective, easy to use, safe, logistically
10
D. Hemostasis
The ability of the body to control the flow of blood following vascular injury is
paramount to continued survival (unknown). Hemostasis is the process of forming clots in the
walls of damaged blood vessels and preventing blood loss while maintain blood in a fluid state
within the vascular system. A collection of complex interrelated systemic mechanisms operates
Events in hemostasis includes (1) contraction of the smooth muscles in the blood vessel
wall. This reduces the flow of blood and loss from the defect in the vessel wall. The term for
The process of blood clotting and then the subsequent dissolution of the clot, following
repair of the injured tissue, is termed hemostasis consisting of (1) the constriction of blood
vessels (vasoconstrictive phase), (2) the clumping together (aggregation) of platelets (platelet
E. Related Studies
The study of Wang (2011) showed that Artemisia annua L. extract and C12 have obvious
pro-coagulant effect in-vitro. C12 is the part of 20% methanol fraction after column
chromatography of MCI gel is the hemostatic active fraction of Artemisia annua L. The crude
extract of Artemisia annua L. has the hemostatic activity, and the R-value is 19.85%, while the
R-value of positive control is 8.54%. Since the ethyl acetate extract isnot completely dissolved
and there is granular which can accelerate the solidification of the plasma in the physic liquor,
the R-value of ethyl acetate extract is a little high than the n-butanol extract. And the result
11
showed that the R-value of n-butanol extract is higher. There was significant effect of the crude
According to the study of Rajasekaran (2010), the significant reduction in bleeding time
suggest that the Eupatorium ayapana leaf extract and fresh juice have positive effect on
hemostatic phase of wound healing and may possibly act on the integrity of blood vessel or
involvement of platelets forming the hemostatic plug. Platelets are the blood cells involved in
coagulation or it may inhibit the formation of prostaglandin by the vessel walls during injury.
Prostaglandin released during injury is responsible for vessel relaxation, which leads to increase
Goker (2008) worked on the hemostatic actions of the folkloric medicinal plant extract
Ankaferd Blood Stopper. It was observed that the addition of ABS to normal plasma and serum
resulted in the very rapid (< 1s) formation of a protein network. ABS- induced protein network
was capable of regulating further coagulation and hemostatic reactions. Routine hemostatic and
biochemical tests have revealed that the ABS-induced network formation depended upon
interactions between ABS and blood proteins, mainly fibrinogen and other proteins possibly via
agglutination of these molecules. The ABS-induced network formation isrelated to the functions
12
CHAPTER III
METHODOLOGY
A. Collection of Plant
Mindanao State University, Marawi City. The whole plant was washed with clean water and
the damaged parts were discarded. Distilled water was used for the final rinse. The plant was
identified using the Flora of Manila book and Pulak Majumder’s Review Article on
The whole plant of Peperomia pellucida was utilized in this study. It was air-dried
for three days, then chopped and pounded using wooden mortar and pestle. The extraction
was done through squeezing method using sterilized cheese cloth. The collected extract was
then filtered twice using filter paper. The brownish liquid filtrate was transferred into
sterilized glass vials and stored in the refrigerator, to prevent contamination, until required.
Appropriate units of solution extracts were made. Distilled water was used as medium
for dilution. There were three solution extracts prepared, 50%, 75% and 100% crude extracts.
Fifty percent solution extract was prepared by dilution of 0.5ml pure extract into 1ml distilled
water. Seventy-five percent solution extract was obtained by diluting 0.75ml pure extract
into 1ml distilled water. One hundred percent extract was a pure concentrated extract. Final
solution extracts were kept in pre-labeled sterilized vials and stored in the refrigerator (Plate
2).
13
C. Screening of Blood Donors
Blood donors were drawn from healthy volunteer donors (n=15) of both genders
(ages 18- 25 years old). The donors were told the purpose of the work and written donor
consent forms (Appendix 4) for blood collection were given to all respondents and signed.
Blood sample was drawn from donor via venipuncture (Appendix 6) method using
syringe at the antecubital area of the forearm (Plate 3). Only approximate 1mL of blood was
drawn from the donor using 1ml sterile disposable syringe with 25-gauge needle size. The
blood drawn was then transferred into pre-warmed glass tubes immediately.
For blood transfer, glass tubes used were sterilized and pre-warmed in a 37o C water
bath. There were four glass tubes designated accordingly for normal plasma, for 50%
extract, for 75% extract and for 100% extract; tubes were pre-labeled as A, B, C, and D
From the collected blood of 1ml, about 0.2ml aliquots of blood were distributed into
four prepared glass tubes. Immediately, appropriate crude extract solutions were added into
respective tubes. Upon immediate addition of crude extracts, stopwatch was started. Clotting
time determination on glass tubes were observed simultaneously by tilting test tubes to a 45o
angle at intervals of 30 seconds until the test tube can be completely inverted without spilling
the contents, which is blood, was completely clotted (Plate 4). Glass tube A contained 0.2ml
14
whole blood (control) and observed for any trace of blood clot. Glass tube B contained 0.2ml
whole blood and received 0.2ml of 50% crude extract and observed for any trace of blood
clot. Glass tube C contained 0.2ml whole blood and received 0.2ml 75% crude extract and
observed for any trace of blood clot. Glass tube D contained 0.2ml whole blood and received
0.2ml 100% crude extract and observed for any trace of blood clot. Observed clotting time
F. Experimental Design
This study used Randomized Complete Block Design (RCBD). The experimental
treatments were basically whole blood distributed into glass tubes. The design had three
Blocks (I, II, and III) with four Treatments (A, B, C, and D) each; and each Treatment had
five replicates totaling to 60 treatments (n=60). All Treatments A were treatments with no
crude extract received (control). All Treatments B received 50% solution extract. All
Treatments C received 75% solution extract. All Treatments D received 100% crude
extract.
(control) (50% crude extract) (75% crude extract) (100% crude extract)
BLOCK I
T1 T1 T1 T1
T2 T2 T2 T2
T3 T3 T3 T3
T4 T4 T4 T4
T5 T5 T5 T5
15
H. Statistical Tool
Results will be expressed as Mean ± Standard Deviation (S. D.). Statistical analysis
Range Test (DMRT) for determination of differences in mean. A value P less than 0.05
16
Chapter IV
evaluated in vitro on human blood as being mixed with its crude extract. This study involved 15
respondents and treatments totaling to 60 (n= 60). There were three appropriate units of crude
extract tested on treatments: 50%, 75% and 100% crude extracts. Raw data on the blood clotting
Table 1 shows the hemostatic effects of the Peperomia pellucida plant crude extracts on
the normal blood clotting time of human plasma using two-way ANOVA.
Table 1. Two-way ANOVA on the effect of P. pellucida on human whole blood clotting time
Total 195.300 59
* Significant
ns-Not Significant
17
Results reveal that the change in the mean scores between different blocks and treatments
on human blood clotting time is significant that not only due to chance (P> 0.05). This shows
that the P. pellucida plant crude extract did induce an effect on the normal clotting time of the
human blood treatments. However, the interaction between the block and the treatment was not
Table 2. DMRT on the effect of P. pellucida on human whole blood clotting time
Subset*
Treatment N 1 2 3
15 a
C- 75% Crude extract 5.9867
15 ab bc
B- 50% Crude extract 7.0067 7.0067
15 c
A- Control Group 7.2200
15 d
D- 100% Crude extract 8.5867
DMRT was used to determine which pairs of treatment means on the clotting time of
human blood were different. Table 2 shows that the 75% Crude extract treatment and the 50%
Crude extract treatment were in the same subset. That is, the clotting time rate of 75% Crude
extract was not significantly different from 50% Crude extract. This subset had the fastest rate
18
of clotting time based on observed means. Similarly, 50% Crude extract was followed by the
Control group in the next subset having the same clotting time range. Significant interval in the
clotting time range in this subset can be attributed to the diluted concentration of 50% Crude
extract that gives a clotting time near equivalent with the Control group. Also, this suggests that
even the lesser concentration of the Crude extract at 75% can still induce blood clotting.
On the other hand, only the 100% Crude extract belongs to the last subset. This implies
that 100% Crude extract was significantly different from the other three treatments. This subset
has the slowest rate of clotting time, in other words it prolongs the normal clotting time human
blood usually takes. Probably, there are chemical components in this P. peperomia plant that
when used at higher concentrations can cause prolongation on the clotting mechanism.
Table 3. Mean values on the effects of P. pellucida on human blood clotting time
Table 3 illustrates the mean values of blood samples between different treatments.
Control standard samples were run parallel and comparatively with the test samples on blood
19
human volunteers as shown in Appendix 2. Control group clotting time was 7.22 minutes. It
was distant to P. pellucida that showed 6.00 minutes for 75%Crude extract/0.2ml of blood; and
near equivalent of 7.00 minutes for 50%Crude extract/0.2ml of blood. Meanwhile, 100%Crude
extract showed anticoagulation for 8.59 minutes before it clotted. The normal average clotting
Results showed imply that the greater the value of the mean, the more clotting time is
prolonged. Thus, it demonstrates that 75% crude extract established appropriate concentration
for coagulation. On the other hand, pure concentration of extract slows down blood coagulation.
20
Chapter V
blood was investigated in vitro. There were three different crude extract concentrations tested,
50%, 75% and 100%. Initially, blood was drawn from fifteen volunteer donors. Hemostatic
effects were measured by determination of blood clotting time using glass tubes noted in
minutes. Blood samples were grouped into three blocks and then divided into four treatments.
Treatment A is the standard control of whole blood and no crude extract received. Treatment B
received 50% crude extract. Treatment C received 75% crude extract and Treatment D received
100% crude extract. All experiments in the control were replicated five times with one blood
donor per replicate. A novel average clotting time of human blood was established locally.
Blood clotting time under different blocks and treatments were compared and analyzed using
Results showed that Peperomia pellucida plant crude extract induced an effect on the
clotting time of human blood at 50% and 75% crude extract concentrations. Moreover, the 75%
crude extract revealed to hasten the normal clotting time while 50% crude extract showed near
equal results with the control group. However, 100% crude extract induced prolongation on the
21
1. Investigate and isolate the potential components of the plant that induces hemostatic
mechanisms in blood.
2. Utilize other blood clotting assay methods such as Dale’s method and Duke’s method.
3. Apply other hemostasis measurement assays like bleeding time assay, prothrombin time (PT)
5. Increase number of replicates in the study to rule out variations caused by genetic and
22
LITERATURE CITED
BOOKS
Rang, H.P., Dale, M.M., Ritter, J.M., (1999). Pharmacology (4th ed.). Churchill Livingstone,
UK:Churchill Livingstone Publishing Company.
Ganong, W.F., (2000). Review of Medical Physiology (10th ed.).Kuala Lumpur, Malaysia:
Khapu Publishing Company.
Ghani A. (1998). Medicinal Plants of Bangladesh (1st ed.). Dhaka, Bangladesh: Asiatic Society
of Bangladesh.
Wagman, R.J. (2000). The New Complete Medical and Health Encyclopedia (Vol. 2, pp. 408
409). New York: Ferguson Publishing Company.
JOURNALS
Bayma, J., Arruda, M., Muller, A., Canto, W. (2008). A Dimeric ArC 2 compound from
Peperomia pellucida. Phytochemistry, 55, 779- 782.
Craig, W.J., (1999). Health promoting properties of common herbs. Am J Clin Nutr, 70, 491
499.
Davison, C., (2006). A biochemical study of the antidiabetic and anticoagulant effects of
Tulbaghia violacea. Nelson Mandela Metropolitan university, 6, 387-393.
Egwuche, R.U., Odetola, A., Erukrainure, O. (2011). Preliminary investigation into the chemical
properties of Peperomia pellucida. Research J. of Phytochemistry, 5, 48-53.
Muñoz, V., Sauvain, M., Bourgy G., Arrazola S., Callapa, J. (2000). A search for natural
bioactive compunds in Bolivia through a multidisciplinary approach: Part III. Evaluation
23
of the antimalarial activity of plants used by Altenos Indians. Journal
Ethnopharmacology, 71, 123-131.
Goker, H., Haznedaroglu, I.C., Ercetin, S., Kirazli, S., Akman, U., Ozturk, Y., Firat, H.C. (2008).
Haemostatic actions of the folkloric medicinal plant extract Ankaferd Blood Stopper.
The Journal of International Medical Research, 36, 163-170.
Khan, A., Rahman, M., Islam, M. (2010). Isolation and bioactivity of a Xanthone Glycoside
from Peperomia pellucida. Life Sciences and Medicine Research, 1, 1-10.
Khan, M.R., Omoloso, A.D., (2002). Antibacterial activity of Hygrophilia stricta and
Peperomia pellucida. Fitoterapia, 73, 251-254.
Omoloso, A.D. (2002). Antibacterial activity of Peperomia pellucida. Fitoterapia, 72, 250-255.
Ragasa, C.Y., Dumato, M., Rideout, J.A. (1998). Antifungal compounds from Peperomia
pellucida. ACGC Chem, Res. Commun., 7, 54-61.
Rajasekaran, A., Kalavani, M., Arlharasivakumar, G., (2010). Hemostatic effect of fresh juice
and methanolic extract of Eupatorium ayapana leaves in rat model. International Journal
of Biological and Medical Research, 1 (3), 85-87.
Sio, O., Cortes, N., Sia, I. (2012). Acute oral toxicity of the freeze-dried aqueous extract of
Peperomia pellucida in mice. Acta Medica Philippina, 5, 43-48.
Wang, B., Sui, J., Yu, Z., (2011). Screening the hemostatic active fraction of Artemisia annua
in vitro. Iranian Journa Pharm,acological research, 1, 243-236.
Winston, J.C. (1999). Health-promoting properties of common herbs. Am. J. Clin. Nutr., 70,
491-499.
24
Unpublished Undergraduate Theses
Ytable, J. (2012). Antibacterial effect of Peperomia pellucida ethanolic extract and its
synergistic effect with standard antibiotics against selected bacteria. Undergraduate
thesis, Mindanao State University, Marawi City.
ELECTRONIC SOURCE
Cao, H.J. (2012). Philippine Medicinal Plants. Retrieved 4 August 2012, from
http://www.stuartexchange.org/index.html
25
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Appendix 1. Raw data for blood clotting time on treatments per blocks
Clotting time
Treatment A Treatment B Treatment C Treatment D
(control) (50% crude extract) (75% crude extract) (100% crude extract)
mins. mins. mins. mins.
BLOCK I
Clotting time
Treatment A Treatment B Treatment C Treatment D
(control) (50% crude extract) (75% crude extract) (100% crude extract)
mins. mins. mins. mins.
BLOCK II
Clotting time
Treatment A Treatment B Treatment C Treatment D
(control) (50% crude extract) (75% crude extract) (100% crude extract)
mins. mins. mins. mins.
BLOCK II
26
Appendix 2. Mean values on effect of P. pellucida on human whole clotting time
A 6.5800 0.54498 5
B 7.0600 0.60663 5
C 5.5200 0.92574 5
I
D 8.9200 1.3976 5
Total 7.0200 1.52371 20
A 8.8400 3.78920 5
B 7.7200 0.97314 5
C 7.0600 1.16962 5
II
D 9.1200 1.78241 5
Total 8.1850 2.21627 20
A 6.2400 0.60663 5
B 6.2400 0.71972 5
C 5.3800 0.69785 5
III
D 7.7200 1.13886 5
Total 6.3950 1.14362 20
27
MINDANAO STATE UNIVERSITY
College of Natural Sciences and Mathematics
DEPARTMENT OF BIOLOGY
Marawi City
January 2013
Ma’am/Sir,
Greetings!
I, Norlainie Castro Omar, a proponent of the research study “Hemostasis effects of Peperomia pellucida
plant extract, in vitro”, would like to indicate my interest on requesting for your participation as a
respondent (Blood donor) in this study. The study concerns the investigation on the usefulness of the
said plant extract against hemorrhage and its effects on the management of blood clotting during
hemostasis.
In line with this, the study shall utilize human blood samples as its experimental variables, which will be
drawn from donors using Syringe Method Venipuncture. Attached herein are the details on how the
proponent shall perform the procedures on blood collection as based from non-surgical procedure
descriptions and guidance. An informed written consent will be given for you to affix your insignia of
volunteer participation.
Rest assured that the proponent would be liable on any unfortunate circumstances during the blood
collection.
Thank you.
Yours truly,
NORLAINIE C. OMAR
Thesis Proponent
Recommended by:
28
INFORMED WRITTEN CONSENT
Date
The study has been well explained to my understanding by the proponent, affixed
herein is my signature that may serve as my voluntary willingness to participate.
Witness Respondent
Date
The study has been well explained to my understanding by the proponent, affixed
herein is my signature that may serve as my voluntary willingness to participate.
Witness Respondent
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MINDANAO STATE UNIVERSITY
College of Natural Sciences and Mathematics
DEPARTMENT OF BIOLOGY
Marawi City
October 9, 2012
Sir:
Greetings!
The undersigned senior student from the Department of Biology is a proponent of the proposed thesis
study entitled “Hemostatic Effects of Peperomia pellucida Plant Crude Extract, in vitro”. She would like
to indicate her interest on requesting you to be the proponent’s thesis adviser. Thus, you shall be
notified for consultations needed and undertakings of the proponent regarding to the said thesis study.
The proponent would be appreciative to have your honor supervising her study. Your expertise and
knowledge in Medical Physiology studies would be of great assistance on the success of this study.
The proponent of the study is hoping for a positive response on this matter.
Thank you.
Respectfully yours,
NORLAINIE C. OMAR
IV BS Biology
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Plate1. Experimental plant Pansit-pansitan (Peperomia pellucida)
Plate 2. Crude extracts of experimental plant. (A) 50% crude extract; (B) 75% Crude extract;
(100%) Crude extract.
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Plate 3. Blood collection via Venipuncture Syringe Method
Plate 4. Blood clots on tubes. (A) Control; (B) 50% crude extract; (C) 75% crude extract; (D)
100% crude extract.
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