UNIVERSITY OF SANTO TOMAS GRADUATE SCHOOL

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PROXIMATE AND PHYTOCHEMICAL COMPOSITION OF

METHANOLIC LEAF EXTRACT OF PIPER BETLE L. (IKMO)

A Thesis Proposal
Presented to the
Graduate School
University of Santo Tomas

In Partial Fulfillment
Of the Requirements of the Course
Pharmaceutical Chemistry 2

by
WYNZI MALLEN; JASON MONTESA; RIZZIEL NEMES; PETER
MICHAEL NOCON; YANCEE OLIVA
October 2014

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ABSTRACT

Phytochemical screening of the methanol extract of Piper betle

leaves revealed that the leaf extract contains alkaloids, terpenes,
anthraquinones, flavonoids, tannins, saponins and steroids.

Keywords: Piper betle, solar systems, universe, aliens,

space

UNIVERSITY OF SANTO TOMAS GRADUATE SCHOOL

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TABLE OF CONTENTS
Page
Prefatory Note (Optional)

1.0 The Problem Rationale

4
Introduction
4
Research Impediments

2.0 The Research Questions

2.1 Literature Review
7
2.2 Research Questions
22
3.0.

The Research Methods

3.1 Methods and Materials
25
3.1.1 Standards, Reagents and Chemicals
26
3.1.2 Extraction (or Sampling) Procedure
27
3.1.3 Ethical Considerations (If Applicable!)
32
3.2 Analysis
3.2.1 Instrumental Analysis
28
3.2.2 Data Interpretation and Calculations
29

References
33
Examples only! (remove those not
applicable)
Appendix I:
Permit to Conduct the Interviews
37
Appendix II:Request for Pertinent Data
38

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Appendix III:
Informed Consent for Research
Participants
39
Appendix IV:
Informed Consent for Research
Participation
Recruited Concept Mapping Participants
40
Appendix V:Survey Questionnaires
41
Appendix VI:
The Interview Protocol
44
Appendix VII:
The Interview Protocol for
Tutors/Reviewers 46
Appendix VIII:
The Interview Protocol
47
Appendix IX:
Concept Mapping Instrument
48
Appendix X:
Documentary Analysis
49
Appendix XI:
Timetable for Research
50
Appendix XII:
Budget Proposal
51
Curriculum Vitae
52

UNIVERSITY OF SANTO TOMAS GRADUATE SCHOOL

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CHAPTER 1
THE PROBLEM RATIONALE

This chapter must be clear and logical in describing succinctly

the trends in the field that render the chosen topic problematic. This
chapter must end with the delineation of the research barriers or
limitations

of

the

study,

(if

possible,

substantial,

ethical,

methodological, and practical) that the candidate may come across in

the conduct of the study. The last two paragraphs must be devoted to
research impediments (barriers) and limitations of the study.
The first paragraph of the paper begins with the first line of the
paragraph indented one-half inch (5-7 spaces). There are no
additional lines added between paragraphsjust the regular double
space that should be maintained throughout the paper and the
References page, if there is one. Do not use block paragraphing
(extra lines between paragraphs, with new paragraphs beginning
flush left without indentation) for academic papers; such format is
increasingly used these days for business letters, but not for
academic or scholarly papers.

UNIVERSITY OF SANTO TOMAS GRADUATE SCHOOL

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The PAGE NUMBER of the first page of EACH CHAPTER must not
appear (and in this template, was blocked with an opaque square).
The succeeding pages MUST show proper page numbers.

UNIVERSITY OF SANTO TOMAS GRADUATE SCHOOL

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CHAPTER 2
THE RESEARCH QUESTIONS

This chapter includes all types of materials reviewed,

conceptual literature which came from books. Related studies both
local and foreign consisting mainly of peer reviewed journals. Your
aim is to show how the present study relates to the existing
knowledge and previous studies in terms of both similarities and
differences. The formal sub-sections for this chapter are;
Be more judicious in making Section headings that are
unnecessary and inappropriate. If it necessitates headings, or you
are writing a long paper in which headings are needed by
convention or to help your reader, follow APA format requirements
as explained in the American Psychological Association [APA]
(2001) Publication Manual.
Conservative scholarly standards in most disciplines require
strict third-person usage in reports of experiments and studies. APA
rules allow for the use of the first person when appropriate, to
avoid the awkwardness of exclusive third-person usage. A detailed
discussion of this issue, maybe found on section 3.09 (Precision
and Clarity, APA (2010) Publication Manual, p.68, 6th edition). The

UNIVERSITY OF SANTO TOMAS GRADUATE SCHOOL

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subsection on Attribution, list examples of undesirable attribution

in third-person usage when describing steps taken in the
Methodology section.
2.1 Review of the Literature
Must be comprehensive in the light of the chosen variables
with a view to surfacing the research blankspots (unexplored area)
and blindspots (conflicting areas in the literature).
2.2 Research Question/s (or Hypotheses)
A research hypothesis is the statement you created when you
speculate upon the outcome of a research or experiment. Every true
experimental design must have this statement at the core of its
structure, as the ultimate aim of any experiment.

UNIVERSITY OF SANTO TOMAS GRADUATE SCHOOL

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CHAPTER 3
THE RESEARCH METHODS

This

chapter

must

include

details

methodology. Points to highlight includes;

of

your

experimental

Description of the

following; (1) research design, (2) study site, (3) selection of samples,
(4) data/outcome measures , data collection procedure, (5) ethical
consideration and (6) data/mode of analysis; and Must be
strongly supported by the appendices that contain the instruments
or corpus of data to be used. There are two formal sub-sections (3.1
and 3.2) which are;
3.1 Methods and Materials
3.1.1 Standards, Reagents, and Chemicals
This section must have detailed description of materials used.
3.1.2 Extraction (or Sampling or Synthesis) Procedure
This section can have either of the 3 titles depending on your
research.
3.2 Analysis
3.2.1 Instrumental Analysis
This section must have detailed description of materials used.
3.2.2 Data Interpretation and Calculations

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In this section you may include your statistical analysis,

description of softwares etc. For example (NOTE: this was taken
from a journal. since you are writing a proposal, the grammar
form MUST BE IN THE FUTURE TENSE)

MS and MS/MS data

searches were performed using a local Mascot server (Mascot 2.2.0,

Matrix Science, London, UK) against the SwissProt database (version
56.8,

410,

518

sequences).

For

MALDIMS

peptide

mass

fingerprinting, a mass tolerance of 50 ppm was allowed. (taken

from Fouillen et al., Analytical Biochemistry 407 (2010) 3443)

10

UNIVERSITY OF SANTO TOMAS GRADUATE SCHOOL

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BIBLIOGRAPHY
Include all literature actually cited in chapters 1 to 3, arrange
entries ALPHABETICALLY (NOT NUMBERED); strictly use APA
style entries. Sample entries for books, journals, book edition
and internet source are shown below in that order;
Baxter, C. (1997). Race Equality in Health Care and Education.
Philadelphia: Ballire Tindall.
Gaudio, J. L., & Snowdon, C. T. (2008). Spatial Cues More Salient Than
Color Cues in Cotton-Top Tamarins (Saguinus oedipus) reversal
learning.

Journal

of

Comparative

Psychology,

122,

441-444.

doi:10.1037/0735-7036.122.4.441
Hyde, J. S.,& Delamater, J. (2008). Human Sexuality (10th ed.) New
York: McGraw-Hill.
Research Initiatives. (n.d.). Retrieved January 11, 2007, from MIT,
Comparative

Media

http://cms.mit.edu/research/index.php

Studies

website,

11

UNIVERSITY OF SANTO TOMAS GRADUATE SCHOOL

Appendix I

Research
Objectives

Timetable for Research

2010
Ju
n

1. Writing of Proposal
2. Collection of Algal
Materials
3. Fractionation of
Polysaccharides
4. Collection of Blood
and
Isolation of PBMCs
5. Cell Viability Assay
6. ELISA Assay for
Cytokines
7. MRNA extraction
8. Chemopreventive
assays
9. Data
Collation/Analysis
10.
Colloquium/Defense

PAGE

Ju
l

2011
Au
g

Se
pt

Oc
t

No
v

De
c

Ja
n

Fe
b

Ma
r

Ap
r

Ma
y

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UNIVERSITY OF SANTO TOMAS GRADUATE SCHOOL

Appendix II

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Budgetary Requirements

SOURCES OF EXPENSES

Quantit
y

Amount
(PhP)

Collection and Fractionation of

Polysaccharides
Collection of Sargassum
80% Ethanol
0.1M HCl
1.25M NaOH
cetyl pyridinium chloride
99% acetic acid
99% ETOH
Lyophilization

1
10 L
2L
2L
100 g
1L
5L
1

10,000
5,765.00
20
210
6000
390
3600
2500
28,485

Blood Collection and Isolation of

PBMCs
vacutainer tubes
Ficoll-Hypaque gradient soln

1 pack
2 packs

700
15,000
15700

100 ml
100 ml
20 ml
100 ml
100 ml
1g
1L
100 ml
2 packs
2 packs
2 packs
1L
IL
100 ml
1L
500 ml
1g

2,126.79
1,745.54
1,600.89
859.82
977.68
7,366.07
3,682
9,511.61
4,625
4, 875
4,968.64
2, 417
5,372
1,449
6,484
902
7,238
35,000
93,909.0
4

Tissue Culture Reagents

L-glutamine
Penicillin-Streptomycin
Hepes Buffer
Sodium bicarbonate
Trypsin 0.25% EDTA
MTT
DMEM
FBS qualified H1
Std. TC flask 50 ml canted neck
15 ml conical tube w cap poly
50 ml conical tube w/ cap pol
Dulbeccos Phosphate Buffered
RPMI 1640
Trypan Blue Reagents
Freezing Medium DMSO Serum free
Sodium Pyruvate Soln
DPPH reagent
Hep G2 Cell Lines

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Budgetary Requirements, continued

RNA Isolation and RT-PCR
reagents
Trizol reagent
RNAse Out Recom. Inhibitor
RT-PCR Mix Reactions
Agarose electro reagents
M Tris Buffer solution
Gel Pilot DNA loading dye
Pipet tips yellow
PCR tube
gloves
Tissues and cleaning reagents
ELISA KIT Reagents
IL-6 Human antibody
IL-1 Human antibody
TNF- Human Antibody

Quantity
100 ml
1 kit
1 set
1
1
1
1

kit
pack
pack
pack

1 kit
1 kit
1 kit

Amount
(PhP)
14,241.07
12,473.21
53,000.00
5,400
7,168
2,880
680
2, 518
600
500
99,460.2
8
48,000
48,000
48,000
144,000

TOTAL
381,554.32

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UNIVERSITY OF SANTO TOMAS GRADUATE SCHOOL

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Curriculum Vitae

Insert
your
picture
here

Limit to one (1) page !

15