EFFECTIVENESS OF CRUDE EXTRACT OF MAYANA LEAVES FOR BLOOD COAGULATION PROCESS An Undergraduate Thesis Presented to the Faculty of the College of Allied Health Sciences Medical Technology Department Cagayan State University Andrew’s Campus, Tuguegarao City In Partial Fulfillment Of the Requirements for the Degree Bachelor of Science in Medical Technology By Rhoe Anne C. Calabazaron Randolph Ryan G. Gasmin Abigail C. Guinucay Jocelyn P. Oledan Niki Valencia D. Pamittan Rachelle I. Tugad March 2012
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EFFECTIVENESS OF CRUDE EXTRACT OF MAYANA LEAVES FOR BLOOD COAGULATION PROCESS
An Undergraduate ThesisPresented to the Faculty of the
College of Allied Health SciencesMedical Technology Department
Cagayan State UniversityAndrew’s Campus, Tuguegarao City
In Partial Fulfillment Of the Requirements for the Degree
Bachelor of Science in Medical Technology
ByRhoe Anne C. CalabazaronRandolph Ryan G. Gasmin
Abigail C. GuinucayJocelyn P. Oledan
Niki Valencia D. PamittanRachelle I. Tugad
March 2012
Chapter I
THE PROBLEM AND ITS BACKGROUND
Introduction
Use of herbs and medicinal plants as the first medicines is a universal phenomenon.
Every culture on Earth, through written or oral tradition, has relied on the vast variety of
healing plants for their therapeutic properties. The majority of herbal products available
today originated from the same traditional formulas or ingredients. The use of herbal
medicines predates human history. Herbal medicine creates a more potent, effective and
efficient treatment to ensure quicker responses to re-establish health and balance.
Most people used herbal medicines to treat their wounds, especially in rural areas.
Some people use herbal perching for the leaves to produce extract and put the leaves on
the wound. They believed that herbal plants are effective in treating wounds especially in
minimizing its bleeding. Not knowing that these herbal plants are also effective in
minimizing the bleeds on wounds which can be used as an immediate treatment for
hemostasis. Herbal medicine can provide a better quality of life
Figure 1. Shows the Independent, Intervening and Dependent Variables on the Effectual Use of the Crude Extract of Mayana Leaves (Coeus Blumei benth) for Blood Coagulation Time in Different Blood Types.
Mayana (Coeus Blumei benth)
- Temperature- Blood type- pH
Blood Coagulation
Cephalin(Positive Control)
Statement of the Problem
The study is designed to find out the effectual use of mayana leaves extract for
blood coagulation.
Specifically, it answers the following questions:
1. What is the medicinal composition of mayana leaves that is contributory to blood
coagulation?
2. What is the extent of effectiveness of mayana leaves extract to hasten blood
coagulation time.
3. What health risk possibilities does mayana leaves extract have to human body
system?
4. Is there a significant difference in blood coagulation time between the four
different blood types using mayana leaves extract?
Assumptions
The assumptions of the study are presented in this from:
1.
Significance of the Study
The study is conducted to find out the effectiveness of mayana leaves extract as to
the blood coagulation. The results of the study are expected to benefit the following:
Remote areas. The researcher’s findings in this study would help the people inform in
remote areas where there is no access to hospitals or clinic that herbal plant is essential to
speed up the blood clot of the lesion. Furthermore, this study would provide additional
option aside from the cephaline or any other commercially available medicine for wound
in cases of emergency situations.
Doctors. The results of this research will aid the doctors to advise what kind of herbal
plant is efficient in treating an abrasion in case of urgent situation.
Public health nurse. The findings of this research will endow additional knowledge
regarding health information given or trained by public health nurse about the
effectiveness of mayana leaves extract to the blood coagulation process.
Researchers. The study would also serve as a proposal and a guide to the future
researchers on what variables they would embrace or focus as they carry out another
study associated to this subject matter.
Scope and Limitation of the Study
The scope of the study is on the effectiveness of crude extract of mayana leaves in
aiding the blood coagulation process. It will be conducted at the laboratory of Cagayan
State University, College of Allied Health Science. The blood types to be used in this
study are decided to be blood type A, B, AB and O. This is to determine the significant
difference of the different blood types on their blood coagulation time when the crude
extract of mayana leaves is used in comparable with cephalin known for its
thromboplastic agent. Since there is some sort of sensitivity in this experimental study,
the accuracy of the result of experimentation varies on the capacity of the researchers,
condition of the blood samples and the materials and instruments that are available to use.
Definition of Terms
The following terms are defined to assist a clear understanding of the study:
Blood clotting. The conversion of fluid blood into a coagulum that involves shedding of
blood, release of thromboplastin, inactivation of heparin, conversion of prothrombin to
thrombin, interaction of thrombin with fibrinogen to form an insoluble fibrin network,
and contraction of the network to squeeze excess fluid.
Blood coagulation. A complex process by which blood forms clots.
Cephalin. A thromboplastic substance which initiates the process of blood coagulation.
Clotting factor. Any substance in the blood that is essential for blood to coagulate.
Crude Extracts. The liquid form of the mayana made by purging of the leaves.
Effectiveness. The fulfilling result of the herbal plants used in the clotting time.
Poller L. et.al.(1972) – Retrieved from http://www.ncbi.nlm.nih.gov/pmc/ articles/PMC477616/ “The Partial Thromboplastin (cephalin) Time Test
Ruggiero F M et. al. (1981)- Retrieved from http://www.ncbi.nlm.nih.gov/ pubmed/7259878 “Comparative Study of Lecithin and Cephalin Exchange between Liposomes and Mitochondria”
Samuel A. Guttman (1944)– Retrieved from http://www.ncbi.nlm.nih.gov/pmc/ articles/PMC435458/pdf/jcinvest00592-0046.pdf “Study on Cephalin-Cholesterol-Flocculation Reaction”
Wadsworth, Augustus et. al (1934) – Retrieved from http://www.jimmunol.org/ content/26/1/25.abstract “Study of the Antigenic Properties of Lecithin and Cephalin”
This chapter will present the research methodology of the study. It includes the
research methodology, materials, equipment and apparatus, procedure training of the
panelists, evaluation of the products and statistical treatment of data.
Research Method
This study uses the descriptive-comparative as experimental design. It describes
the effectiveness of the crude extract of mayana leaves when used in blood coagulation
process. It also correlates the accurate blood clotting time using cephalin as positive
control, the crude extract of mayana leaves as the experimental specimen and the normal
blood clotting time as the negative control. The experimental research controls the
condition of the study. Parallel-group design is the type of experimental design used.
Wherein the normal coagulation time of blood serves as control in comparing the
effectiveness of the crude extract of mayana leaves with the effect of cephalin when used
by different blood types which are A, B, AB and O in coagulation process.
Materials
The materials to be used in this proposed study will be 70% alcohol, crude extract of
mayana, cephalin, A, B, AB, O blood types. Table 1 presents the materials used in the
extraction of crude extract from the mayana leaves (Coeus Blumei benth) and used in the
coagulation time.
Table 1
Materials Quantity
70% alcohol
crude extract of mayana
cephalin
Blood types:
A (3 samples)
B (3 samples)
AB (3 samples)
O (3 samples)
60ml
240μL
240μL
0.18cc
0.18cc
0.18cc
0.18cc
Table 1. Materials used in the extraction of crude extract from the mayana leaves (Coeus
Blumei benth) and used in the coagulation time.
Apparatus
The apparatuses to be used in this proposed study will be clean glass slides, cotton,
disposable lancet, needle, sterile gauze, stopwatch, mortar and pestle, beaker and knife.
Table 2 presents the apparatus used in the extraction of crude extract from the mayana
leaves (Coeus Blumei benth) and used in the coagulation time.
Table 2
Apparatus Quantity
Clean glass slides
Cotton
Disposable lancet (sterile)
Needle
Sterile gauze
Stopwatch
Mortar and pestle
Beaker
Knife
36 pieces
1 pack
36 pieces
36 pieces
1 piece
3 units
1 set
1 piece
1 piece
Table 2. Apparatus used in the extraction of crude extract from the mayana leaves (Coeus
Blumei benth) and used in the coagulation time.
Procedures
The procedure that will be used in conducting the study for coagulation time or
clotting time when the crude extract of mayana leaves is used in different blood types is
called the Slide Method or Drop Method. The steps or procedures in this study will be
done following in this order: (1) Disinfect site of puncture with 70% alcohol sponge and
dry. (2) Puncture to a depth of 3mm. (3) Wipe off first drop of blood. (4) Start the timer
as soon as the second drop of blood appears. (5) Transfer the second drop of blood onto
the center of glass slide. (6) Pass the tip of needle through the drop of blood every thirty
seconds and note for the formation of fibrin strands. (7) Stop the timer as soon as fibrin
strands are seen clinging at the tip of the needle.
The normal value of blood coagulation time in this test is said to be two to four (2-
4) minutes. After conducting the test to the four different blood types, the result will be
noted weather there are differences in the coagulation time of blood types A, B, AB and
O. The time noted will be used as a negative control on the next procedure where testing
the effectivity of mayana leaves in coagulation process while cephalin will be used as
positive control.
Repeat procedure one to five. (6) When the second drop of blood is onto the center
of the glass slide, get a 10μL of crude extract of mayana leaves and mix it with the
specimen. (7) Pass the tip of needle through the drop of blood with a crude extract every
thirty seconds and note for the formation of fibrin strands. (8) Stop the timer as soon as
fibrin strands are seen clinging at the tip of the needle. Then repeat the same procedure
for test of cephalin.
After noting all results in every procedures of this experiment. Tabulate the data
for better comparison. The researchers will write their conclusion on the effectiveness of
crude extract of mayana leaves in blood coagulation time based from the experiment they
conducted.
To obtain a crude extract from mayana leaves. The researchers will do these
following procedures. (1) Collect the mayana leaves early in the morning, before the heat
of the day. Choose mature leaves that are fully formed and mature but are not aged or
damaged. (2) Wash the mayana leaves with clean water. (3) Finely chop the leaves with
the knife to easily release all the components from the plant. (4) Pound the leaves using a
mortar and pestle to make extract. (5) Place the crude extract in the beaker. The container
should be cleaned, dried and sterilized with boiling water before use for a pure extract.
Figure 2.1
Crude extract of mayana leaves
Figure 2.1 Process flow sheet for the extraction of crude extract from the mayana leaves (Coeus Blumei benth) that will be used as experimental material for the study.
Wash the mayana leaves with clean water
Pound the leaves using a mortar and pestle to make extract.
Collect the mayana leaves early in the morning. Choose mature leaves that are fully formed.
Finely chop the leaves with the knife.
Place the crude extract in the beaker.
Figure 2.2
Blood Type A Blood Type B Blood Type AB Blood Type O
Figure 2.2 Process flow sheet for the comparison of the four different blood types (A, B, AB, O) as the crude extract of mayana leaves and cephalin are used in the coagulation time, comparing the result with the natural clotting time of the blood.
Natural Blood Clotting time
(- control)
Crude extract of mayana leaves(Experimental)
Cephalin(+ control)
Disinfect site of puncture with 70% alcohol sponge and dry
Puncture to a depth of 3mm
Wipe off first drop of blood
Start the timer as soon as the second drop of blood appears
Transfer the second drop of blood onto the center of glass slide
Pass the tip of needle through the drop of blood every thirty seconds and note
for the formation of fibrin strands
Stop the timer as soon as fibrin strands are seen clinging at the tip of the