Mark Andrew Maray
$6/hr
Virtual Assistant
- Reply rate:
- 50.0%
- Availability:
- Full-time (40 hrs/wk)
- Age:
- 24 years old
- Location:
- San Fernando City, Pampanga, Philippines
- Experience:
- 0 years
Angeles University Foundation
Bachelor of Science in Medical Technology
In Vitro Determination of the
Anticoagulant Activity of
Terminalia catappa (Talisay)
Leaf Extract in Human Blood
Presentation of Research Proposal
GROUP 2 - MT3B
RESEARCH ADVISER
Maray, Mark Andrew M.
Villanueva, Ella Mae D.
Ramos, Maura Gale G.
Canlapan, Mary Shane M.
Gorospe, Fatima Bianca M.
Bayani, Amiela Grace G.
Ungcad, Francis Dave D.
Dr. Annalyn Navarro
MARAY
I. INTRODUCTION
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Increase in mortality rate due to cardiovascular diseases, venous thromboembolism, heart
failure, and embolic stroke -- Worldwide concern
Compelling therapy continues to be a challenge and presents the need for new forms of
management to improve the impacts of hemorrhage
Coagulation - Blood supply is blocked and the flow of blood is reduced
At present, the control of the blood clotting process involves drug interventions to
prevent further tissue impairment
1
GOROSPE
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
I. INTRODUCTION
Anticoagulants - Interact with the natural blood coagulation system of the body,
manage abnormal blood clotting mechanisms, reduce fibrin formation
Aspirin, Prozac, Paxil, Heparin, and Warfarin
Adverse effect: Bleeding
2
GOROSPE
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
I. INTRODUCTION
Medicinal plants
Alternative to drug anticoagulants
Proven to be sources of anticoagulant molecules (Coumarin, Flavonoids)
Influence the blood vessels and the circulation of blood
Therefore, the prospects of obtaining a novel compound have become higher.
3
GOROSPE
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
I. INTRODUCTION
Terminalia catappa
Tropical almond or Talisay
The leaves possess vitamins
A, B, and C, as well as
carbohydrates, proteins, fat,
and fiber.
Phytochemical
properties
:
flavonoids, tannins, saponins,
phytosterols,
glycosides,
cyanogenic,
Appendix A.
T. catappa's tree and leaf
alkaloids,
phenols, and steroids
4
VILLANUEVA
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
I. INTRODUCTION
Terminalia catappa
Hence, this study would like to challenge that Talisay leaf extract will also
exhibit anticoagulation properties in human blood.
5
VILLANUEVA
I. INTRODUCTION
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Significance:
This study aims to determine if Terminalia catappa has anticoagulant
activity in human blood. If T. catappa is proven to have anticoagulant
activity, it could be used as an alternative for existing anticoagulant
medications.
6
VILLANUEVA
II. OBJECTIVES
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
General Objective
This research will be conducted to determine the anticoagulant activity of Terminalia catappa
(Talisay) leaf extract in human blood
Specific Objective
To determine the anticoagulant activity of Terminalia catappa leaf extract in different
concentrations (25%, 50%, 75%, 100%)
7
GOROSPE
III. MATERIALS AND METHODS
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Research Design and Locale
Research Design
True Experimental Research
Research Locale
Room 107, Medical Technology Laboratory, Saint Lorenzo Building,
Angeles University Foundation
8
CANLAPAN
III. MATERIALS AND METHODS
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Conceptual Framework
9
CANLAPAN
III. MATERIALS AND METHODS
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Study Participants
Exclusion Criteria
Refusal to give informed consent
Sample Size
Active smoker or currently using any
15 participants (Ayodele, Onajobi,
tobacco products and e-cigarette
& Osoniyi, 2019)
History of major bleeding or trauma
within 6 months prior to signing informed
Inclusion Criteria
consent
18-35 years old without any disease
Propensity to bleeding
Free from dyslipidemic disorders
Allergic reaction to latex
No history of non-steroidal antiinflammatory drug (NSAID) use
Have not taken any medication that
can alter blood coagulation one (1)
week prior to the conduct of
research
10
MARAY MARAY
III. MATERIALS AND METHODS
Research Procedure
Plant Leaves Collection
T. catappa leaves - Km 77 McArthur
Highway, City of San Fernando, Pampanga
Healthy and mature leaves - largest w/o
visible morphological abnormalities
Certification of leaves at the Bureau of
Plant Industry, Department of Agriculture,
City of San Fernando, Pampanga
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Leaf Extraction
1. Leaves were oven-dried at 75°C for 4 hours and ground into fine
powder.
2. Soaked in 70% ethanol for 48 hours @ RT.
3. Filtered using Whatman No.1 filter paper.
4. Rotary evaporation to further dissolve solvent.
5. Waterbath at 40°C to concentrate.
6. Dried extract is refrigerated in air-tight container at 20°C until use.
7. To reconstitute, the ff. values were used:
Leaf Extract
Concentration
Volume of NSS
Weight of Plant
Extract
25 g/mL (%)
10 mL
2.5 grams
50 g/mL (%)
10 mL
5 grams
75 g/mL (%)
10 mL
7.5 grams
100 g/mL (%)
0 mL
10 grams
11
RAMOS
III. MATERIALS AND METHODS
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
20-minute WHOLE BLOOD CLOTTING TEST (WBCT20)
Bedside and widely used in operations to
monitor
reversal
of
heparinization
after
cardiac surgery
Simple and effective by utilizing a small
amount of blood to define clot formation
observed directly and is stable within 20
minutes
12
RAMOS RAMOS
III. MATERIALS AND METHODS
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Research Procedure
Preparation of the Test Samples and Performance of WBCT20
1mL of human blood for each AC concentration (0.11mL):
25% (25g/mL)
50% (50g/mL)
75% (75g/mL)
100% (100g/mL)
The 0.11mL:1mL (leaf extract to human blood) is computed using
the standard 1:9 ratio of AC to blood.
Inverted five (5) times to allow proper mixing of sample and leaf extract &
left to sit at room temperature for 20 minutes
Lamb et al. (2021)
PRESENCE OF CLOT = NEGATIVE AC ACTIVITY
NO CLOT = POSITIVE AC ACTIVITY
13
RAMOS RAMOS
III. MATERIALS AND METHODS
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Research Procedure
Preparation of Blank, Positive and Negative Controls
For positive control, the researcher used vacutainer tube with light blue
top containing sodium citrate as anticoagulant
Precise amount of blood needed depends on the vacutainer used
Inversion of three to four times
For the negative control, 1mL of blood is transferred to a glass tube
without any reagent or leaf extract
For the blank, 0.11mL of NSS is transferred to a glass tube and 1mL of
blood is then added
All controls are left to sit at room temperature for 20 minutes
Controls are used to test if the participants' blood have a normal clotting
activity
14
MARAY MARAY
III. MATERIALS AND METHODS
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Informed
Consent
Process,
Duration of Participation, and
Withdrawal Criteria
Ethical Considerations
Angeles University Foundation – Office of
Interview > Blood collection > Analysis
the Vice President for Research and
Purely voluntary so subjects can withdraw at any time
during this study
Innovation Ethics Review Committee
15
UNGCAD
III. MATERIALS AND METHODS
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Risks and Inconveniences
Privacy, Confidentiality, and
Data Management
Pain from venipuncture
Needle phobia; profuse sweating, paleness,
nausea, allergic reaction from latex allergy, and
fainting
The researchers are Medical Technology students
supervised by the research adviser
Proper patient aftercare will be performed
Collected informations shall only be used for research
purposes and kept confidential
Practices are approved by the Ethics Review Committee
(ERC)
Data collected identifying the participants will not be
utilized outside this research
Authorization statement must be signed
Before publication, all identifying information is eliminated
and authorization from the participants is acquired
Research information of the participants data shall be
disposed or destroyed after examined and investigated
Benefits of the Study
For the participants:
Future reference for the potential of the leaf
extract
as
an
alternative
choice
for
anticoagulants
Findings will be conveyed to the participants
16
UNGCAD
IV. DATA ANALYSIS
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Statistical Tool
Interval data
Simple frequency and percentage
Results and Interpretation
Clot formation after 20 minutes: Negative
No clot formation after 20 minutes: Positive
17
BAYANI
V. RESULTS
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Table 1. Comparison of Anticoagulant Activity of Ethanolic
Extract of Talisay Leaves With Different Concentrations in
Human Blood
25g/mL
50g/mL
75g/mL
100g/m
L
Frequency and
Percentage (With Clot)
7
(46.67%)
2
(13.33%)
2
(13.33%)
0 (0%)
Frequency and
Percentage (No clot)
8
(53.33%)
13
(86.67%)
13
(86.67%)
15
(100%)
Total
15
(100%)
15 (100%)
15
(100%)
15
(100%)
GOROSPE
V. RESULTS
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Table 2. Comparison of Anticoagulant Activity of the Control
Group in Human Blood
Positive Control
Negative Control
Blank
Frequency and Percentage
(With Clot)
0 (0%)
15 (100%)
15 (100%)
Frequency and Percentage
(No clot)
15 (100%)
0 (0%)
0 (0%)
Total
15 (100%)
15 (100%)
15 (100%)
BAYANI
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
VI. DISCUSSION
25g/mL
50g/mL
75g/mL
100g/mL
Frequency and Percentage (With Clot)
7 (46.67%)
2
(13.33%)
2 (13.33%)
0 (0%)
Frequency and Percentage (No clot)
8 (53.33%)
13 (86.67%)
13 (86.67%)
15 (100%)
Total
15 (100%)
15 (100%)
15 (100%)
15 (100%)
CANLAPAN
VI. DISCUSSION
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Phytochemical Components of Terminalia catappa
Tannins and Flavonoids present in the leaf extracts can
interfere with platelet aggregation, clotting factor activation
Mainly Clotting factors (FXa FIIa) - Bijak et al. (2014) (29)
UNGCAD
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
VII. CONCLUSION
25 g/mL
+/-
Thus, Talisay ethanolic extract has
anticoagulant activity, and different
50 g/mL
minimal +/-
75 g/mL
minimal +/-
anticoagulant activities but a 100
100 g/mL
+
g/mL concentration yields complete
Control group exhibited the expected results
concentrations showed significant
no clot formation
Negative = clot formation
Positive = no clot formation
VILLANUEVA
VII. RECOMMENDATION
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
Study is limited to:
WBCT20
one of the basic methods in defining clot formation
Researchers recommend:
Medical Clearance
Other Hematological Methods
Prothrombin Time
Activated Partial Thromboplastin Time
Use other parts of the Talisay Tree
bark
branch
limb
root
Other solvent
Ethyl acetate
Methanol
MARAY
MARAY
V. REFERENCES
In Vitro Determination of the Anticoagulant Activity of
Terminalia catappa (Talisay) Leaf Extract in Human Blood
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Life Sciences. 2021 May;10(2):231.
2. Ayodele OO, Onajobi FD, Osoniyi O. In vitro anticoagulant effect of Crassocephalum crepidioides leaf methanol extract and fractions on human blood. Journal of experimental pharmacology. 2019;11:99.
3. Borela VT, Balunsat KA, Briones AJ, Briones PN. In-Vitro study of the anticoagulant property of Terminalia catappa (Talisay) leaf extract using Gallus gallus (Chicken) Blood. Journal LA Lifesci. 2020;01(03): 025029. doi:-/journallalifesci.v1i3.158
4. Pawlaczyk I, Czerchawski L, Kuliczkowski W, Karolko B, Pilecki W, Witkiewicz W, Gancarz R. Anticoagulant and anti-platelet activity of polyphenolic-polysaccharide preparation isolated from the medicinal
plant Erigeron canadensis L. Thrombosis Research. 2011;127(4):328-40.
5. Dayaganon AJ, Sarpong SM, Sapilan JB, Fernandez AM. Anticoagulant activity of Horseradish Moringa oleifera and Oregano Coleus aromaticus leaf extracts and Kamias Averrhoa bilimbi fruit juice on human
blood samples. Optima. 2013;1:11-.
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7. Castillo RR, Atilano AA, David-Ona DI, Napiza-Granada C, Cruz-Sevilla MR, Torreblanca H, et al. May Measurement Month 2017: An analysis of blood pressure screening in the Philippines—south-east asia and
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10. Abebe W. Review of herbal medications with the potential to cause bleeding: dental implications, and risk prediction and prevention avenues. The EPMA Journal [Internet]. 2019 Jan 8;10(1):51–64. Available from:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC-/
11. Thomson L, Evans B. Terminalia catappa (Tropical Almond). Species Profiles for Pacific Island Agroforestry [Internet]. 2006 Jan;2(2):1–20. Available from:
https://www.researchgate.net/publication/-_Terminalia_catappa_Tropical_Almond
12. Fraenkel JR, Wallen NE, Hyun HH. How to design and evaluate research in education. New York: Mcgraw-Hill Humanities/Social Sciences/Languages; 2012.
13. Abdel-Razek, TT, Binnaser, HAH. Smoking and platelet function. International Journal of Advances in Pharmacy, Biology and Chemistry [Internet]. 2014 Jan. Available from:
https://www.researchgate.net/profile/Tarek-Abdel-Razek/publication/-_Smoking_and_Platelet_Function/links/56284a8b08ae04c2aead8664/Smoking-and-Platelet-Function.pdf
14. Benjamin JM, Chippaux J-P, Sambo BT, Massougbodji A. Delayed double reading of whole blood clotting test (WBCT) results at 20 and 30 minutes enhances diagnosis and treatment of viper envenomation.
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https://statistics.laerd.com/statistical-guides/one-way-anovastatistical-guide.ph
16. Lamb T, Abouyannis M, De Oliveira SS, Shenoy K R, Geevar T, Zachariah A, Sharma SK, Bhatt N, Mukaka M, Harriss E, Lalloo DG. The 20-minute whole blood clotting test (20WBCT) for snakebite coagulopathy—
A systematic review and meta-analysis of diagnostic test accuracy. PLoS neglected tropical diseases. 2021 Aug 10;15(8):e-. Patten ML, Newhart M. Understanding Research Methods. 10th ed. New York: Routledge Taylor & Francis Group; 2018
17
Angeles University Foundation
Bachelor of Science in Medical Technology
THANK YOU FOR
LISTENING!
GROUP 2 - MT3B
Maray, Mark Andrew M.
Villanueva, Ella Mae D.
Ramos, Maura Gale G.
Canlapan, Mary Shane M.
Gorospe, Fatima Bianca M.
Bayani, Amiela Grace G.
Ungcad, Francis Dave D.
RESEARCH ADVISER
Dr. Annalyn Navarro
