1
Antioxidant Activity and Phytochemical Content of Cooked and
Uncooked Forms of Allium ascalonicum L. (shallot)
A Research Presented to the
Special Health Sciences Senior High School
De La Salle Medical & Health Sciences Institute
City of Dasmarinas City, Cavite
In Partial Fulfillment of the Requirements
for the course Practical Research 1
Ababao, Sophia Mikaela. M
Ancheta, Ma. Patricia Eunice T.
Arana, Alliyah Mariel G.
Arraz, Pauline Therese F.
Javier, Regine Viviene R.
July 2019
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THESIS/RESEARCH OUTPUT OPEN ACCESS PERMISSION
I/We, Sophia Mikaela M. Ababao, Ma. Patricia Eunice T. Ancheta, Alliyah Mariel G.
Araña, Pauline Therese F. Arraz and Regine Viviene R. Javier, author(s) of the
thesis/research output entitled “Antioxidant Activity and Phytochemical Content of
Cooked and Uncooked Forms of Allium ascalonicum L. (shallot)”, submitted to De La
Salle Medical and Health Sciences Institute Special Health Sciences Senior High school
Department in partial fulfilment of the degree/year level/residency in Grade 12, do
hereby grant DLSMHSI the right to reproduce, publish and distribute copies of the said
thesis or research output in whatever form deemed appropriate by DLSMHSI subject to
applicable laws, institutional policies and contracts. Specifically, the following rights are
granted to DLSMHSI:
1. To upload a copy of this thesis or research output and be made accessible online
through the institutional repository or database,
2. To publish a copy of this thesis or research in the Compendium of Research
Abstracts or similar publications, whether print or online,
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is/are properly acknowledged and the work is cited.
Permission is given for the following people to have access to this thesis or research
output:
Available to the general public
YES
Available only after consultation with author/thesis adviser/Dean or Director
YES
Available only to those bound by confidentiality agreement
YES
SOPHIA MIKAELA M. ABABAO
PAULINE THERESE F. ARRAZ
Researcher
Researcher
MA. PATRICIA EUNICE T. ANCHETA
REGINE VIVIENE R. JAVIER
Researcher
Researcher
ALLIYAH MARIEL G. ARAÑA
EVELYN D. SEGOVIA, LPT, MA
Researcher
Class Adviser
VIVIAN B. RAMIREZ, MSc
Director, SHSSHS
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ABSTRACT
Allium ascalonicum L. (shallot), a member of the family Liliaceae, has
been generally used as a food ingredient like Allium cepa and Allium sativum L..
But apart from being a staple ingredient in the kitchen, the bulb of the shallot is
also
being
used
as
an
antihelminthic,
anti-inflammatory,
antiseptic,
antispasmodic, and carminative herbal herbal medication. One of the goal of the
researchers is to determine specific phytochemicals in Allium ascalonicum L. that
may be responsible for its medicinal property. The ethanolic extract from the
sample will be subjected to flavonoids and phenol testing. Another objective in
conducting this study is to determine the possible antioxidant activity of Allium
ascalonicum L. in both cooked and uncooked forms and to identify which among
the two forms contain the highest flavonoids and phenolics content using DPPH
Assay. In determining the significant difference of the different concentrations of
cooked and uncooked forms of Allium ascalonicum L., results showed that the
cooked sample with a concentration of 1000µg/mL and uncooked sample with a
concentration of 500µg/mL contain the highest amount of antioxidant activity
wherein the cooked sample with a concentration of 250µg/mL and uncooked
sample with a concentration of 1000µg/mL showed the lowest antioxidant
activity.
Keywords: Allium ascalonicum L., phytochemicals, ethanolic extract,
antioxidant activity, highest flavonoids and phenolics content, DPPH assay,
cooked and uncooked forms
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APPROVAL SHEET
The capstone research entitle “Antioxidant Activity and Phytochemical
Content of Cooked and Uncooked Forms of Allium ascalonicum L. (shallot)”,
prepared and submitted by Sophia Mikaela M. Ababao, Ma. Patricia Eunice T.
Ancheta, Alliyah Mariel G. Araña, Pauline Therese F. Arraz and Regine
Viviene R. Javier, in partial fulfilment of the requirements for Practical Research
I, has been examined and recommended for acceptance and approval for Research
Colloqium.
VIVIAN B. RAMIREZ, MSc
Thesis Adviser
Research Colloqium Panel
Approved by the committee on Research Colloqium with a grade of
EVELYN D. SEGOVIA, LPT, MA
Member, RRP
MYRA MICHELLE M. MOJICA
Member, RRP
Approved and accepted in partial fulfilment of the requirements for Special
Health Sciences Senior High School Program.
VIVIAN B. RAMIREZ, MSc
Director, Special Health Sciences Senior High School Program
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ACKNOWLEDGMENT
The proponents would like to express their warmest and sincerest gratitude
to the following people who brought a big contributions to this research, thus
making it possible and successful.
Miss Vivian B. Ramirez, their research adviser for her dedication in giving
support and guidance as they gradually accomplish this research.
Sir John Paul Pascua from DLSMHSI’s faculty, for his patience and
understanding as he provided the researchers his effort through his guidance and
supervision.
Sir Lance Tan, from where they obtained major guidance from, for his
enthusiasm and immense knowledge on the field of biochemistry, along with his
enormous effort in making our research possible.
Lastly, to the Almighty Father for always giving them strength especially
through the hardships in making this research.
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TABLE OF CONTENTS
Title Page
1
Thesis/Research Open Access Permission
2
Abstract
3
Approval Sheet
4
Acknowledgement
5
Table of Contents
6
CHAPTER 1 INTRODUCTION
Background of the Study
9
Conceptual Framework
13
Statement of the Problem
14
Objectives of the Study
15
Scope and Limitations
16
Significance of the Study
17
Definition of Terms
19
CHAPTER 2 LITERATURE REVIEW
21
CHAPTER 3 METHODOLOGY
Research Design
26
Research Setting
27
Materials and Equipment
28
7
Research Procedure
29
Data Gathering Procedure
31
Statistical Treatment of Data
33
Data Analysis
34
CHAPTER 4 RESULTS AND DISCUSSION
35
CHAPTER 5 SUMMARY, CONCLUSION AND
RECOMMENDATION
Summary
46
Conclusion
48
Recommendation
50
Cited References
52
Appendices
A. Letter to the OIC Director of Bureau of Plant Industry
57
B. Certificate of Verification of Plant Sample
58
C. English Editing Certification
60
D. Time Table of Activities (Gantt Chart)
62
E. Budgetary Requirements
63
F. Photo Documentation
64
Curriculum Vitae
71
8
List of Tables
Table 1. Materials, Subjects, Reagents, Equipment
28
Table 2. Antioxidant activity of uncooked samples
39
Table 3. Antioxidant activity of cooked samples
40
Table 4. Summary of Results
44
List of Figures
Figure 1. Framework of the Study
13
Figure 2. Qualitative Phytochemical Screening for Flavonoids
35
Figure 3. Qualitative Phytochemical Screening for Phenols
37
Figure 4. Free radical scavenging activity of samples
41
Figure 5. Total Flavonoids Content of Cooked and
42
Uncooked samples
Figure 6. Total Phenolics Content of Cooked and
Uncooked samples
43
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INTRODUCTION
Background of the Study
Plants are rich in phytochemical components. Phytochemicals occur naturally
in medicinal plants, leaves, vegetables and roots. These phytochemicals, also known as
phytoconstituents, are natural bioactive compounds that are present in plants which plays
a significant role in the defense mechanism of humans. The reason why plants,
specifically fruits and vegetables, are recommended to its fullest consumption is that it
helps in making the body strong and free from diseases. (Wadood, 2013)
Phytochemicals are divided into two: (1) Primary Constituents which includes
sugars, amino acids, proteins and the chlorophyll. (2) Secondary Constituents which
includes alkaloids, flavonoids, phenolics, saponins and more. Alkaloids are present in
morphine and caffeine wherein it acts as an analgesic substance. Flavonoids are widely
known for its anti-allergic, anti-inflammatory, antioxidant, and antimicrobial effect.
These are found on almost all fruits and vegetables which also reduce the risk for heart
diseases and cancer. Phenolics are present in some of medicinal products like lozenges
and ointments. Saponins, on the other hand, are present in beans and legumes which help
protect the body from cancers and it also helps in lowering the blood cholesterol level.
(Afolabi, 2013)
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Phytochemical analysis is a process used to separate and identify the
medicinally active portions of plant tissues using solvents and reagents. In the process of
extraction, the solvents will reach the solid part of the plant and will eventually dissolve
with compounds of similar polarity. The compounds present in plants are approximately
composed of complex mixtures of plant metabolites, either in liquid, semisolid or powder
as the solid form which are intentional for oral or external use. (Kumar, 2011)
There are three (3) basic indicators that affects the quality of an extract:
1. Plant part used
2. Solvent or reagent used for extraction
3. Extraction procedure
Plant parts can vary from leaves, bark, roots, fruits, flowers, seeds and such.
Any part of a plant may contain bioactive compounds. The plant part sample can either
be fresh or dried for the extraction of some additional plant components. But dried plants
are better for the reason that several plant tissues differ in water content.
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Choosing of solvents is a crucial part, because the success of the determination
of the active compounds greatly depends on this. These are the properties to consider a
solvent a good one: low toxicity, ease of evaporation at low heat, rapid physiologic
absorption of the extract, preservative action, and inability of the extract to disintegrate.
There are five (5) common solvents that are widely used in extraction: Water, Acetone,
Alcohol, Chloroform and Ether. (Tiwari, 2011)
Variations in plant extraction may arise due to the following: duration of the
extraction period, choice of solvent, pH of the solvent, temperature, and the solvent-tosample ratio. The basic principle is to grind the plant material (wet or dry) as pure as it
can be, which causes the surface area for extraction to expand as well as its rate. Previous
studies recorded that the ideal solvent-to-sample ratio is 10:1. Some of the different
variations of extraction methods or techniques have been already mentioned earlier.
(Tiwari, 2011)
Allium ascalonicum L., also known as shallot, is a member of the family
Liliaceae. It has been generally used as a spice or as a food ingredient just like Allium
cepa and A. sativum L. during the prehistoric times. The bulb of the shallot has an
antihelminthic, anti-inflammatory, antiseptic, antipasmodic, carminative, diuretic,
expectorant, febrifuge, hypoglycemic and hypotensive properties. It helps in lowering the
risk of heart attack and accumulation of plaque in the arteries. Just like Allium cepa, it is
also used in speeding up the healing process of wounds.
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Shallot are rich in minerals, vitamins, and trace elements. They also contain
potassium, phosphorus, manganese, magnesium, copper and iron. They are also rich in
vitamin B6 (pyridoxine), which plays an integral role in functioning of the nervous and
immune systems. Shallots are a source of vitamin B9 or folic acid as well, which is
essential to the cell division and DNA synthesis and contributes to red cell formation.
Shallots also provide vitamin C and vitamin A, the two major vitamins that have multiple
health benefits. They are also an outstanding source of selenium, a trace element that
defend cells against aging and contributes to healthy skin and hair. Together with
Vitamin E, also found in shallots, selenium contributes to the formation of antibodies that
protect the body against microbes and viruses.
The aim of this study is to determine the phytochemical content present in both
cooked and uncooked forms of Allium ascalonicum L. (shallot) using various test to
analyse and compare which is more prevalent in amount between these two most
commonly found phytochemicals. This will also help in determine the antioxidant
activity of cooked and uncooked forms of Allium ascalonicum L. (shallot) and how it is
beneficial to the human body.
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Conceptual Framework
Cooked and Uncooked Allium
ascalonicum L. (shallot) extract
Phytochemical screening that will determine the compounds present
in Allium ascalonicum L.
Rate of Antioxidant activity
present in Cooked and Uncooked
Allium ascalonicum L. (shallot)
Phytochemical Content present
in Cooked and Uncooked Allium
ascalonicum L. (shallot) extract
extract
Figure 1. Framework of the Study
Figure 1 will be adapted from the procedures that were described by Trease and Evans
(1989) which shows the variable considered in the study. The Allium ascalonicum L.
(shallot) extract, both cooked and uncooked, constitute the input which is the problem
regulated in this study. To determine the compounds, present in the Allium specie,
phytochemical screening will serve as the method for collecting the data. And lastly, the
output of the study is the rate of antioxidant activity and the phytochemical content found
in cooked and uncooked forms of Allium ascalonicum L. (shallot).
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Statement of the Problem
This study aims to determine the antioxidant activity and the phytochemical
content present in cooked and uncooked forms of Allium ascalonicum L. (shallot) through
phytochemical screening and analysis.
General Problem: What is the antioxidant activity and phytochemical content of cooked
and uncooked forms of Allium ascalonicum L. (shallot) extract?
Specific Problem:
1. What is the antioxidant activity of Allium ascalonicum L. (shallot) in:
a. cooked form?
b. uncooked form?
2. Which among both cooked and uncooked forms of Allium ascalonicum L.
(shallot) contain the highest:
a. flavonoids content?
b. phenols content?
3. Is there a significant difference among the varying concentrations of cooked
and uncooked Allium ascalonicum L. (shallot) ethanolic extracts as an
antioxidant in terms of:
a. 2,2 Diphenyl-1-picrylhydrazyl (DPPH) Assay?
b. Total Flavonoids Content?
c. Total Phenolics Content?
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Objectives of the Study

To determine the antioxidant activity of Allium ascalonicum L. (shallot) in both
cooked and uncooked forms.

To determine which among the cooked and uncooked forms of Allium
ascalonicum L. (shallot) contain the highest flavonoids and phenols content.

To determine if there is a significant difference among the varying
concentrations of cooked and uncooked Allium ascalonicum L. (shallot)
ethanolic extracts and ascorbic acid as an antioxidant in terms of 2,2 Diphenyl1-picrylhydrazyl (DPPH) Assay, Total Flavonoids Content and Total Phenolics
Content.
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Scope and Limitations
The scope of this study mainly focuses on finding the antioxidant activity of
cooked and uncooked forms of Allium ascalonicum L. (shallot). The proponents are also
determined to find out the phytochemical content but will only give emphasis on its
flavonoids amount and phenolic content as they are of simpler structure compared to
other chemicals.
The genus Allium is composed of onion and garlic scented herbs that are
mostly found in almost all parts of the world. This concludes that the selection of the
researchers’ species will be used in the experiment is primarily because of its availability.
This study is only limited to using Allium ascalonicum L. (shallot) primarily because it is
one of the most commonly grown species here in the Philippines. There are various tests
which could help determine the chemical substances found in Allium ascalonicum L.
(shallot), but researchers decided to use the most available, low-cost, least time
consuming and accurate chemical substances to achieve the objectives set.
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Significance of the Study
Allium ascalonicum L. (shallot) in different ways like for industrial purposes
and household uses. It is a significant ingredient with multiple purposes. Thus, knowing
the rate of its antioxidant activity as well as the phytochemical content present through
phytochemical analysis and DPPH Assay would help in identifying the certain health
benefits and nutrients an individual could get.
Medical Field. This study will contribute to the existing knowledge on the medical field
and will also inform certain medical professionals regarding the difference on the rate of
antioxidant activity of cooked and uncooked forms of Allium ascalonicum L. (shallot).
Knowing what is more beneficial could help them inform their patients properly.
Nutritionists and Dietitians. This study will inform nutritionists and dietitians on the
components found in Allium ascalonicum L. (shallot) most especially the antioxidant
activity of cooked and uncooked shallots. This could help them promote healthy eating
habits and organize a nutrition plan that is rich with the antioxidants leading to prevention
of certain health complications of patients.
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Teachers. This study will inform the teachers about the existence of the abundant
phytochemicals and what are their beneficial contributions to individual health. Teachers
could be an instrument on instilling awareness and influence their students in prioritizing
foods that are rich in phytochemicals such as the allium species. This would help them
decrease the risks in acquiring diseases such as cardiovascular diseases and cancer.
Parents. This study will help parents in considering allium species as an abundant part of
the meals they prepare for their child/children. Knowing the phytochemical components
would let them prioritize the right intake appropriate for their child’s health. This could
also inform them that phytochemicals could be an alternative remedy if their child has a
certain deficiency.
Future Researchers. This will help the future researchers to have an idea and a plan
towards establishing the research. Furthermore, they can adapt the ideas given by the
proponents of the study. This will serve as guide to be able to fill-in information that the
researchers were not able to provide.
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Definition of Terms
Abundant. A substance or a given object that has an excess value or is available in a
large quantity.
Bioactive Compounds. These are the extra-nutritional constituents in small quantities of
food that has a corresponding effect.
Chemical Substance. Possesses a constant composition and cannot be separated into
components; can be simple or compound; can be seen in nutrients such as protein,
carbohydrates, and fat which is present in food.
Constituents. An element that is considered as portions of something that makes up a
whole; a part of something whole.
Distinct. Distinguished as different and is easily identified that is present from a similar
type.
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Nutritional Value. Defines what are the contents of the food and the impacts of
constituents in the body relating to carbohydrates, protein, cholesterol, fat, etc.
Phytochemicals. Compound of nutrients that are produced by plants (can be found in
fruits, vegetables, etc.) in order to produce or carry out biochemical activity when
consumed which is believed to protects cells from being damaged that could lead to
cancer.
Reagents. A substance or a compound that is being added to a food sample in order to
determine the chemical composition by observing the reaction and identifying the present
nutrients.
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REVIEW OF RELATED LITERATURE
In this chapter, related studies about phytochemicals, benefits and various tests
of allium species are presented which came from the in-depth search done by the
researchers. To broaden and improve this study, previous researches were read and
reviewed.
A research published by Block (2010) entitled “The chemistry of garlic and
onions” states that the Allium sativum L. (garlic) that belong to the Lilly family which has
a strong odor and taste that makes it a unique flavoring agent on household use. Its
regular consumption could lead to health benefits such as its antiatherosclerotic effect
which stabilizes good cardiovascular conditions through lowering cholesterol levels and
blood pressure. However, high amount of intake or exposure to skin could lead to burning
and irritation due to its strong, burning effect. Second, another study published by
Galeone & Tavani, et al. (2009) entitled “Allium vegetable intake and risk of acute
myocardial infarction in Italy” states that the Allium cepa L. (onion) also belongs to the
Lilly family, which is one of the major vegetable crops produced by marginal farmers. It
is one of the main source of household and medicinal applications. High consumption
could lead to various benefits such as the decrease in blood sugar levels and decrease the
risk of heart complications. Lastly, a study established by Cragg & Newman (2008)
entitled “Plants as a source of anti-cancer agents.”. The Allium ascalonicum L. (shallot) is
a member of the Liliaceae family and a variety of Allium cepa (onion) which has been
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used mainly as a spice traditionally from the ancient times. It has many benefits including
bacterial and fungal prevention. It could also be a potential anti-inflammatory and anticancer treatment such as preventing the growth of malignant tumors.
In a research conducted by Arora & Sharma, et al. (2017) entitled
“Phytochemical analysis and evaluation of antioxidant potential of ethanol extract of
Allium cepa and ultra-high homeopathic dilutions available in the market: A comparative
study”, qualitative phytochemical analysis requires reagents to react with the bioactive
chemical substances in plants. Different tests were studied by various researchers in order
to find the most accurate and efficient way of phytochemical analysis. The following are
the tests used in the said research:
Salkowski reaction is used to determine if there are steroids present in the plant.
Chloroform is added, followed by concentrated H2SO4. The solution of red-brown color
tests positive for steroids.
Alkaloids can be tested in four ways: Dragendorff’s test, Hager’s test using picric
acid, Mayer’s test using Potassium mercury iodide in H2O, and Wagner’s test. HCl will
be used to dissolve the plant samples then the following reagents will be added.
Dragendroff’s test will form red-brown depositions of solids in the solution, Mayer’s test
will form a cream-colored precipitate, Hager’s test will form yellow solids, and Wagner’s
test will form a reddish brown precipitate.
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Tannins can be tested in two ways: Ferric chloride test which will show blue and
green colors and Lead acetate test which will form white depositions of solids in the
solution which measures positive for Tannins.
Flavonoids can be tested using two ways: Alkaline reagent test that will form an
intense yellow color to near transparent after dilution of HCl, and the Shinoda test which
will show a crimson red color as indication of flavonoid presence.
Saponin glycosides can be tested by the Froth Formation test in which the sample
will be diluted to 20mL of distilled water then be shaken vigorously. If a centimeter layer
of foam appears stable for ten minutes, then it indicates the presence of saponin
glycosides.
Amino acids are tested by the Ninhydrin test in which a solution of ninhydrin in
ethanol will be added to the sample. Purple color will be shown as indication of presence
if amino acids.
Testing Cardiac Glycosides uses the Keller-Killiani test. The sample solutions are
added with glacial acetic acid, FeCl solution, and pure H2SO4. The solutions will create a
blue color which indicates the presence of Cardiac Glycosides.
Hydroxyanthraquinone test is used to test for Anthraquinone Glycosides wherein
potassium hydroxide will be combined to the sample. A red color from the mixed
solutions indicates positive for Anthraquinone Glycosides.
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Proteins will be tested using the Biuret test. Copper sulfate and Sodium
hydroxide are added to the sample solution and are mixed thoroughly. The indication of
proteins shows a purple color from the solution.
In a research produced by Suru (2018) entitled “Onion and garlic extracts lessen
cadmium-induced nephrotoxicity in rats”, states that because of the doubts about the
safety and efficacy of chelating agents, a search for alternative, affordable and safer
therapy in medicinal plants were made which drew attention to the functional health
effects of Allium cepa L. (onion) and Allium sativum L. (garlic). Onion and garlic are
widely accepted, versatile vegetables because of their medicinal and culinary properties.
Onions are rich in flavonoid while garlic is primarily subsidized with organosulfur
compounds.
A study established by Dixon & Blumenthal (2013) entitled “Principles and
Practice of Phytotherapy”, states that the phytochemical compound flavonoids are
widespread and are very common in the plant kingdom. It functions as plant pigments
and is responsible for the colors of other flowers and fruits. Flavonoids are commonly
consumed in human diet because of its abundancy in plants. On the other hand, tannins
are defined as vegetable substances capable of tanning animal hides to produce leather.
They are polyphenolic compounds like Flavonoids that have an affinity for proteins.
Another research by Gazuwa & Makanjuola (2013) entitled “The Phytochemical
Composition of Allium cepa / Allium sativum and the Effects of Their Aqueous Extracts
(Cooked and Raw Forms) on The Lipid Profile and the other Hepatic Biochemical
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Parameters in Female Albino Wistar Rats” stated that there is a higher level of cholesterol
in both forms of aqueous extracts of garlic and onions. However, the aqueous extracts
coming from the cooked form of the vegetables showed a higher magnitude than raw
extracts. Triglycerides are composed of three fatty acids esterified to a glycerol backbone
and are transported in the blood as core constituents of all lipoproteins. Measuring the
tryglyceride level is very useful in the identification of primary and secondary
hyperlipoproteiemia, dyslipidemia and tryglyceridimia.
Also, it was also stated that both Allium cepa L. (onion) and Allium sativum L.
(garlic) contain very important medicinal values. Onions are comprised of the most
important flavonoids namely quercetin derivatives which help modify the diabetic status
in cells and animal models. It is reported that regular intake of onions can lower the
cholesterol levels and thus prevent the incidence of atherosclerosis and diabetic heart
disease. This was speculated to be effected by the presence of the organosulfur
compounds which lower the homocysteine levels and risk factors for heart attacks and
strokes. On the other hand, garlic constitutes of sulfur containing compounds. The
chemical components in each plant is said to be active principles in drugs such as allicin
(a component of garlic) in antibiotics, organo-sulfur compounds in onion believed to
confer antioxidant, anti-inflammatory, anti-allergic and antithrombic. However, these
active principles can be affected by the various temperature leading them to lose their
medicinal significance.
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METHODOLOGY
In this chapter, specific procedures and techniques with slight modifications
were used as a research strategy to analyse the data and information of cooked and
uncooked forms of Allium ascalonicum L. (shallot).
3.1 Research Design
The researchers employed an experimental research method.
According to Blakstad (2008), experimental research is a type of research which
uses manipulation and controlled testing to fully comprehend causal actions. It is usually
conducted with a scientific approach, and a set of variables are kept constant while the
other set of variables are being manipulated by the researchers. It is also known to be the
one of the founding quantitative research methods.
In this study, experimental research method sought to determine a relationship
between the rate of antioxidant activity and the phytochemical content found in cooked
and uncooked forms of Allium ascalonicum L. (shallot). The use of various chemicals and
tests determined the rate of antioxidant activity and the phytochemical content in cooked
and uncooked forms of Allium ascalonicum L. (shallot).
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3.2 Research Setting
The verification of Allium ascalonicum L. (shallot) was conducted at the Crop
Research & Production Support Division of the Bureau of Plant Industry in Malate,
Manila, Metro Manila.
The phytochemical extraction and analysis of Allium ascalonicum L. (shallot) was
conducted at the Chemistry Laboratory in Lourdes E. Campos Building of De La Salle
Medical and Health Sciences Institute in Dasmariñas City, Cavite.
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3.3 Materials and Equipment
Cooked and uncooked extracts of the specimens was dried using the Heidolph
rotary evaporator.
Materials, Subjects, Reagents and Equipment
Consumables
Equipment
Allium ascalonicum L. (shallot)
Beakers
Aluminum chloride
Flasks
Ascorbic Acid
Funnels
95% Ethanol
Heidolph rotary evaporator
1% Ferric Chloride solution
Lab-grade drying oven
Folin-Ciocalteu’s phenol reagent
Muslin cloth
Sodium acetate
Pipettes and pipette tips
7% Sodium carbonate
Test tubes
Sodium hydroxide
Test tube rack
Sterile distilled water
UV-Visible Spectrophotometer
Table 1. Materials, Subjects, Reagents and Equipment
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3.4 Research Procedure
1. Allium ascalonicum L. (shallot) was collected from SM Molino in Bacoor City on
March 2019.
2. Sample of the specimen was verified in Bureau of Plant Industry on March 2019.
3. The bulb of the sample was washed with sterile distilled water.
4. The outer covering of the bulb was manually removed and peeled off, then
rewashed with sterile distilled water.
5. The bulb was weighed then cut into small parts and squashed.
6. Sample of raw shallots was cooked for 5 minutes and then stored at room
temperature.
6. 95% Ethanol was added to the squashed preparation and macerated for 24 hours
with interval shaking.
7. Muslin cloth was used to filter the samples.
8. The filtrated extract was then collected using the Heidolph rotary evaporator.
9. Test for flavonoids was done by conducting the Alkaline Reagent test in which the
ethanolic extracts of the samples was mixed with a few drops of Sodium Hydroxide
solution.
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10. Test for phenols was done by conducting the Ferric Chloride test in which the
ethanolic extracts of each samples was treated with 1% Ferric Chloride solution.
11. Tests of each of the chemical constituents from all the samples was then observed
to conclude the qualitative phytochemical analysis of Allium ascalonicum L. (shallot).
12. The cooked and uncooked ethanolic extracts was dried using a lab-grade drying
oven.
13. To determining the rate of antioxidant activity, 2,2 Diphenyl-1-picrylhydrazyl
(DPPH) Assay, Aluminum chloride colorimetric method and Folin-Ciocalteu’s
method was used.
14. Data obtained from the experiment was recorded and verified.
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3.5 Data Gathering Procedure
After all variables was verified and extracted, the researchers proceeded to
their data gathering procedure. Several tests were done in order for them to gain the
data they need.
3.5.1 Screening for flavonoids
The Alkaline Reagent test was done by treating 3mL of ethanolic extract with
a few drops of Sodium Hydroxide solution. The formation of an intense yellow color,
which became colorless with the addition of the dilute Hydrochloric acid, indicated the
presence of flavonoids.
3.5.2 Screening for phenols
The Ferric Chloride test was done adding a few drops of Ferric Chloride in
3mL of ethanolic extract with 3mL of water in a test tube. The formation of a darkcolored fluid indicated the presence of phenols.
32
3.5.3 Determination of Rate of Antioxidant Activity
3.5.3.a 2,2 Diphenyl-1-picrylhydrazyl (DPPH) Assay
The antioxidant activity of Allium ascalonicum l. (shallot) was determined
using the DPPH free – radical scavenging method. 0.5mL of extract was mixed with
1.5mL of 0.1 milliMolar DPPH solution in absolute ethanol and was set aside for 1
hour. The reduction of the DPPH radical was determined by measuring the absorbance
of the solution at 517 nm using a UV-Visible Spectrophotometer.
The radical
scavenging was calculated as the percentage inhibition of DPPH discoloration using
the equation:
% 𝐼𝑛ℎ𝑖𝑏𝑖𝑡𝑖𝑜𝑛 = [−1 (
|𝐴𝑠𝑎𝑚𝑝𝑙𝑒 − 𝐴𝑣𝑖𝑡𝑎𝑚𝑖𝑛 𝑐 |
)] 𝑥 100
𝐴𝑣𝑖𝑡𝑎𝑚𝑖𝑛 𝑐
3.5.3.b Total Flavonoids Content
The total flavonoid content was conducted using the aluminium chloride
colorimetric method in which the 0.2mL of ethanolic extract was diluted to
concentrations of 0µg/mL, 40µg/mL, 80µg/mL, 120µg/mL, 160µg/mL and 200µg/mL.
The samples were further diluted with 0.6mL ethanol and was treated with 0.04mL
aluminium chloride and 0.04mL of sodium acetate. The solutions were mixed with
1.12mL of sterile distilled water. The flavonoids content was read using the UVVisible spectrophotometer at 415nm and was expressed using mg Quercetin
equivalence per gram.
33
3.5.6.c Total Phenolics Content
The total phenolics content was determined using the Folin-Ciocalteu's
method. Part of the 0.1mL cooked and uncooked samples was diluted to
concentrations of 0µg/mL, 40µg/mL, 80µg/mL, 120µg/mL, 160µg/mL and 200µg/mL.
Then, it was further diluted with 1.5mL of sterile distilled water. The solutions were
treated with 0.1mL Folin-Ciocalteu's phenol reagent and was shaken. After 10
minutes, 0.3mL of 20% Na2CO3 solution was added to each mixture. The mixtures
were incubated at 40° Celsius for 20 minutes. The total phenolic content was read
using the UV-Visible spectrophotometer at 755nm and was expressed as Gallic Acid
Equivalents (GAE)/g of crude.
3.6 Statistical Treatment of Data
Statistical analysis was performed using Statistical Tool for Agricultural
Research (STAR). After determining the antioxidant activity, total flavonoids and
phenolics content, each sample was analysed using one-way Analysis of Variance
(ANOVA) and Tukeys’ Honest Significant Difference. This was done with a 95%
level of confidence.
34
3.7 Data Analysis
The results for the qualitative phytochemical screening was arranged based
on the presence of phytochemicals found Allium ascalonicum L. (shallot). Each extract
was mixed with a respective reagent under room temperature with different procedures
and time duration. As for the analysis of results, a change in the color of the solution
indicated that the cooked and uncooked samples have a positive result. Whereas,
absence of change in color indicated that the cooked and uncooked samples have a
negative result.
Also, the laboratory test results for determining the rate of antioxidant
activity as well as the total flavonoids and phenolics content of cooked and uncooked
forms of Allium ascalonicum L. (shallot) was tabulated then graphed for further
analysis. (see Chapter 4)
35
RESULTS AND DISCUSSION
This chapter presents the analysis of the results and data collected from the
phytochemical screening and determination of the rate of antioxidant activity of
the crude ethanolic extract of cooked and uncooked forms of Allium ascalonicum
L. (shallot).
4.1 Phytochemical Screening
The presence of the selected phytochemicals, namely flavonoids and
phenolics were determined using the ethanolic extract of the cooked and
uncooked samples of Allium ascalonicum L. (shallot).
Cooked
Uncooked
Figure 2. Qualitative Phytochemical Screening for Flavonoids
Figure 2 shows that the cooked and uncooked samples of Allium
ascalonicum L. (shallot) has an indication of the presence of a certain metabolite,
flavonoids.
36
The Alkaline Reagent test, when performed, will give an intense yellow
color which will become colorless with the addition of the acid. Correspondingly,
the results showed an intense yellow color which then became transparent after
adding dilute Hydrochloric acid. The results indicated that there is a presence of
flavonoids in the cooked and uncooked extract.
The presence of flavonoids in the crude ethanolic extract of both cooked
and uncooked forms of Allium ascalonicum L. (shallot) is an indicator of the
potential antioxidant activity of the extract. As elaborated by Banjarnahor (2014),
the best-described antioxidant property of flavonoids is derived from their
capability to chelate free radicals immediately by donating a hydrogen atom or by
single-electron transfer by reactive oxygen species.
37
Cooked
Uncooked
Figure 3. Qualitative Phytochemical Screening for Phenols
The cooked and uncooked samples of Allium ascalonicum L. (shallot)
were screened using the Ferric Chloride test. A formation of a dark colored liquid
indicates the presence of phenols in an extract. Results of the presence of phenolic
compounds acquired a positive result in both the cooked and uncooked samples
by turning into dark colored liquids.
The presence of phenols in both cooked and uncooked forms of Allium
ascalonicum L. (shallot) is also an indicator of the potential antioxidant activity of
the extract. A research stated by Minatel & Borges et.al (2017), phenolic
compounds consists of one or more aromatic rings that are attached to hydroxyl
groups. The capability of these compounds to contain antioxidant properties are
related to its hydroxyl groups and phenolic rings. Even with the presence of
antioxidant activity, they have many other beneficial effects such as that is
reduces the risk of having infectious and degenerative diseases.
38
Statement of the Problem no. 1
What is the antioxidant activity of Allium ascalonicum L. (shallot) in cooked and
uncooked form?
4.2 Antioxidant Activity
The antioxidant activity of Allium ascalonicum L. (shallot) was
determined on each of the cooked and uncooked samples with the use of DPPH
radical scavenging assay. Ascorbic acid was used as a positive control of the
DPPH Assay since it is relatively stable. Four variations of concentrations were
used in order to determine if different concentrations of the extract is relevant to
its antioxidant activity. 250µg/mL, 500µg/mL, 750µg/mL, and 1000µg/mL were
the concentrations used for the cooked and uncooked samples, with each of the
concentrations conducted in a triplicate analysis. The results of the DPPH Assay
for each samples were near the rate of antioxidant activity as compared to
ascorbic acid at 0.1mg/mL.
39
Concentrations
Yield (%)
Average yield (%)
93.55957
250µg/mL
93.93114
93.95591
94.37701
94.07976
93.93114
500µg/mL
94.10453
94.3027
93.93114
93.85682
750µg/mL
93.5348
92.81645
93.03939
1000µg/mL
91.9247
92.64305
92.96507
Table 2. Antioxidant activity of uncooked sample
Table 2 shows the antioxidant activity of uncooked Allium ascalonicum L.
(shallot). The average from the triplicate analysis of uncooked samples with
250µg/mL concentrations were 93.95% with an error of 0.409%; 500µg/mL were
94.10% with an error of 0.187%; 750µg/mL were 93.53% with an error of
0.623%; and 1000µg/mL were 92.64% with an error of 0.623%.
40
Concentrations
Yield (%)
Average yield (%)
82.93287
81.89249
250µg/mL
80.55487
82.18974
92.44488
90.51276
500µg/mL
88.72925
90.36413
93.33664
92.71736
750µg/mL
92.22195
92.59351
92.81645
1000µg/mL
92.5192
92.69259
92.74214
Table 3. Antioxidant activity of cooked samples
The mean of the triplicate analysis of the cooked samples with 250µg/mL
concentrations were 81.89% with an error of 1.21%; 500µg/mL were 90.51%
with an error of 1.86%; 750µg/mL were 92.72% with an error of 0.567%; and
1000µg/mL were 92.69% with an error of 0.154%. The cooked extracts only
showed high antioxidant activity when the concentrations were set to 500µg/mL
or higher whilst using a lower concentration of the extracts yield a significantly
lower antioxidant activity.
41
%Antioxidant Activity Relative to
0.1mg/mL Ascorbic Acid
Free Radical Scavenging Activity of
Samples
100
95
a
a
a
ab
b
ab
ab
7
8
90
85
c
80
75
70
1
2
3
Uncooked Allium ascalonicum L.
4
5
6
Cooked Allium ascalonicum L.
Figure 4. Free radical scavenging activity of samples
Figure 4 shows the radical scavenging activity against DPPH in uncooked
samples showed a significantly higher antioxidant capacity as a result compared
to the cooked samples of Allium ascalonicum L. (shallot). To interpret, adding a
significant amount of heat, such as cooking, to Allium ascalonicum L. (shallot)
destroys the antioxidants of the said plant.
42
Statement of the Problem no. 2
Which among both cooked and uncooked forms of Allium ascalonicum L.
(shallot) contain the highest flavonoids and phenols content?
Total Flavonoid Content of Samples
Flavonoid Content in Quercitin
Equivalents (μg/mL )
160
135.36
140
120
100.06
100
80
60
40
20
0
1
Uncooked Allium ascalonicum L.
2
Cooked Allium ascalonicum L.
Figure 5. Total flavonoid content of cooked and uncooked samples
The total flavonoid content of the cooked and uncooked samples were
determined using a Quercetin equivalent (QE). The average yield of the raw
samples for the total flavonoid content was 100.06µg/mL with an error of 7.42
whereas the average yield of the cooked samples was 135.36µg/mL with an error
of 8.84. Shown in the bar graph above, the average yield of cooked samples were
higher than that of the raw samples. This suggests that cooking may allow for the
release of flavonoids that are trapped within the plant’s cells but the heat was not
able to degrade the flavonoids.
43
Total Phenolic Content of Samples
Phenolic Content in Gallic Acid
Equivalents (μg/mL )
300
267.47
250
200
150
109.69
100
50
0
1
Uncooked Allium ascalonicum L.
2
Cooked Allium ascalonicum L.
Figure 6. Total phenolic content of cooked and uncooked samples
The total phenolic content of each samples were identified by triplicate
analysis by using the standard curve of Gallic acid and the amount was expressed
as Gallic Acid Equivalent (GAE) in µg/mL. The average yield of the uncooked
samples for the phenolic compounds was 267.46µg/mL with an error of 4.61
whereas the average yield for the cooked samples was 109.69µg/mL with an error
of 2.77. As seen in the figure above, the total phenolic content of the uncooked
samples yield a higher value than that of the cooked samples. Similar to the
DPPH radical scavenging assay, adding a significant amount of heat, like
cooking, to Allium ascalonicum L. (shallot) destroys the phenolic compounds in
the plant.
44
Statement of the Problem no. 3:
Is there a significant difference among the varying concentrations of cooked and
uncooked Allium ascalonicum L. (shallot) ethanolic extracts as antioxidant in terms of
2,2 Diphenyl-1picrylhydrazyl (DPPH) Assay, Total Flavonoids Content and Total
Phenolics Content?
Concentrations
Mean
N group
Cooked 250µg/mL
81.89
c
Cooked 500µg/mL
90.51
b
Cooked 750µg/mL
92.22
ab
Cooked 1000µg/mL
92.69
ab
Uncooked 250µg/mL
93.69
a
Uncooked 500µg/mL
94.10
a
Uncooked 750µg/mL
93.53
a
Uncooked 1000µg/mL
92.64
ab
Table 4. Summary of Results
Table 4 shows the summary of results for the varying concentrations of cooked
and uncooked samples. Samples with the same letters are not significantly different
from each other. However, the cooked sample with a concentration of 250µg/mL is
evident to be of significant difference among other samples, whether cooked or
uncooked. The table also presents how the cooked sample with a concentration of
45
1000µg/mL and uncooked sample with a concentration of 500µg/mL contain the
highest amount of antioxidant activity. Whereas the cooked sample with a
concentration of 250µg/mL and uncooked sample with a concentration of 1000µg/mL
showed the lowest antioxidant activity.
46
SUMMARY, CONCLUSION AND RECOMMENDATION
SUMMARY
In this study, the Antioxidant Activity and the Phytochemical Content of
Cooked and Uncooked forms of Allium ascalonicum L. (shallot) were screened,
measured, and examined. Through the use of screening for Flavonoids and
Phenols, the researchers were able to determine the total Flavonoids and total
Phenols content of the ethanolic extract of the cooked and uncooked samples
using the Alkaline Reagent test for the presence of Flavonoids and Ferric Chloride
test for the presence of Phenolics.
The rate of antioxidant activity was performed after the phytochemical
analysis of the plant Allium ascalonicum L. (shallot), conducted in accordance to
standard protocol. Through the use of the free radical assays DPPH (2,2
Diphenyl-1-picrylhydrazyl), the researchers were able to determine the
antioxidant activity in cooked and uncooked Allium ascalonicum L. (shallot).
Ascorbic acid was used as a positive control of the DPPH assay and four
variations of concentrations were used in order to determine if different
concentrations of the extract is relevant to its antioxidant activity, with each of the
concentrations conducted in a triplicate analysis.
47
Lastly, the researchers were then able to apply the data they have collected
for statistical analysis. Through the use of Quercetin equivalent (QE), they
determined that the average yield of cooked samples were higher than that of the
raw samples. This suggests that cooking may release of flavonoids that are
trapped within the plant’s cells but the heat was not able to degrade the
flavonoids. Furthermore, through the use of Gallic Acid Equivalent (GAE), they
also found out that the total phenolic content of the raw samples yield a higher
value than that of the cooked samples.
Different factors such as shallot’s phytochemical constituents, free radical
scavenging activity contributed to the conclusion that Allium ascalonicum L.
(shallot), has the potential to be an effective antioxidant, whether cooked or
uncooked, but most specifically when it is uncooked.
48
CONCLUSION
The researchers were able to address the research questions based upon the
data that were gathered and analysed. The following conclusions were achieved:
1. The phytochemical screening for flavonoids through the Alkaline Reagent test
showed that both cooked and uncooked forms of Allium ascalonicum L. (shallot)
was positive for the presence of Flavonoids. Thus, researchers concluded that
both forms contain a certain antioxidant property.
2. The phytochemical screening for Phenols through the Ferric Chloride test
shows that both cooked and uncooked forms of Allium ascalonicum L. (Shallot)
was positive for the presence of Phenols. Thus, similar to the interpretation for
Flavonoid screening, researchers concluded that both forms contain a potential
antioxidant activity.
3. The concentration of 500 µg/mL for uncooked samples have the highest
average yield for antioxidant activity which is 94.10% with an error of 0.187%.
On the other hand, the concentration of 750 µg/mL has the highest average yield
for antioxidant activity which is 92.72% with an error of 0.567%.
49
4. Based on the percentage yield of the four concentrations for cooked and
uncooked, it is conclude that the uncooked forms of Allium ascalonicum L.
possess higher antioxidant activity since its percent yield is higher than the
cooked. Therefore, cooking or exposure to heat could alter and destroy the
antioxidant property of the plant.
5. For the total flavonoid content, the average yield of the raw samples was
100.06µg/mL with an error of 7.42. Whereas, the average yield of the cooked
samples was 135.36µg/mL with an error of 8.84. To conclude, cooking or
exposure to high temperature triggers the presence of flavonoids in Allium
ascalonicum L. (shallot).
6. For the total phenolic content, the average yield of the raw samples was
267.46µg/mL with an error of 4.61. Whereas, the average yield for the cooked
samples was 109.69µg/mL with an error of 2.77. In summary, it is concluded that
phenols are highly present in normal temperatures rather than when the plant is
cooked or exposed to high temperature.
50
RECOMMENDATION
In consideration to the results gathered and analysed by the researchers,
the following recommendations have been formulated to improve and expand the
field of research regarding the phytochemical compounds.
Further research may be conducted regarding the use of the other parts of
the plant Allium ascalonicum L. (shallot), particularly in the stem and roots, for
these may contain other phytochemical compounds not present or detected in the
bulb. Additional investigation of related plants from the family Liliaceae
regarding their antioxidant activities may also prove fruitful, as well as the
determination of the exact components that contribute to the antioxidant
properties of Allium ascalonicum L. (shallot). The relationships between plants
from the same family may be expounded and explored. Changes to the
experimental design such as the utilization of different assay protocols such as
FRAP, ABTS and other established or developing antioxidant assays may also be
useful, while deeper analysis of Allium ascalonicum L. (shallot)’s total phenolic
and flavonoid content, may contribute synergistically with other compounds in
enhancing antioxidant activity. This would help in expanding the field of
knowledge regarding this plant and its capabilities. In addition, the correlation of
the amount of phenols and flavonoids in relation to the antioxidant activity of
both cooked and uncooked forms of Allium ascalonicum L. (shallot) can also be
added in order to further explain the relationship of these variables.
51
Conducting these types of tests is very costly and time-consuming to say
the least. Majority of the chemicals and equipment required to conduct the tests
are expensive and inaccessible. Also, research on phytochemical analysis and
antioxidant activity of a plant require higher level of knowledge that were not yet
fully attained by Senior High School students.
52
References
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(4) Blakstad, O. (2008). Experimental Research. Retrieved Jan 17, 2019 from
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(5) Block, E. (2010). The chemistry of garlic and onions. Scientific American; 252
(March): 114119
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(6) Cragg, G.M. & Newman D.J. (2005). Plants as a source of anti-cancer agents.
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(7) F, A. F. (2013). Phytochemical Constituents of Some Medicinal Plants in South
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(8) Ferrari. “Curcumin: A Culinary Herb and Its Health Benefits.” Journal of Modern
Medicinal Chemistry, 2013, doi:10.12970/2308-8044.2013.01.01.1.
(9) Galeone, C. & Tavani, A., et al. (2009). Allium vegetable intake and risk of acute
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Nutr. 2009
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doi:
10.1007/s00394-008-0771-2. Epub 2009 Jan 13
(10) Gazuwa, S.Y. & Makanjuola, K.H., et al. (2013). The Phytochemical
Composition of Allium Cepa / Allium Sativum and the Effects of Their Aqueous
Extracts (Cooked and Raw Forms) on The Lipid Profile and the other Hepatic
Biochemical Parameters in Female Albino Wistar Rats. Asian J. Exp. Biol. Sci. Vol
4 (3) 2013: 406-410
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(11) Leedy,P. & Ormrod, J. (2016) Practical Research planning and design. Boston:
Pearson.
(12) Minatel, I., & Borges, C., et.al. (2017). Phenolic Compounds: Functional
Properties, Impact of Processing and Bioavailability. IntechOpen, doi: 10.5772/66368
(13) Meriga, B., Mopuri, R., & Muralikrishna, T. (2012). Insecticidal, antimicrobial
and antioxidant activities of bulb extracts of Allium sativum. Asian Pacific Journal of
Tropical Medicine, 5(5), 391-395. doi:10.1016/s1995-7645(12)60065-0
(14) Suru, S. M. (2008). Onion and garlic extracts lessen cadmium-induced
nephrotoxicity in rats. BioMetals, 21(6), 623-633. doi:10.1007/s10534-008-9148-5
(15) Tiwari, P. & Kumar, B., et. al (2011). Phytochemical screening and Extraction: A
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(16) Wadood, A. (2013). Phytochemical Analysis of Medicinal Plants Occurring in
Local Area of Mardan. Biochemistry & Analytical Biochemistry, 02(04).
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(17) Experimental Research- Definition, Types of Designs and Advantages. (2018).
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from
56
APPENDICES
57
APPENDIX A
Letter to the OIC Director of Bureau of Plant Industry
58
APPENDIX B
Certificate of Verification of Plant Sample
59
60
APPENDIX C
English Editing Certification
61
62
APPENDIX D
Time Table of Activities (Gantt Chart)
APPENDIX E
63
BUDGETARY REQUIREMENTS
Items
Total
Allium ascalonicum L. (shallot)
₱ 150.00
Equipment
₱ 410.00
Reagents
₱ 6,140.00
Total Amount
₱ 6,700.00
APPENDIX F
64
Photo Documentation
Preparation of Squashed Samples
Squashed Uncooked Allium ascalonicum l. (shallot)
65
Ethanolic Extraction of Samples using a Rotary Evaporator
Standard Ascorbic Acid for DPPH Assay
66
Cooked 250µg/mL
Cooked 500µg/mL
67
Cooked 1000µg/mL
Uncooked 250µg/mL
68
Uncooked 500µg/mL
Uncooked 750µg/mL
69
Uncooked 1000µg/mL
Quercetin Concentrations
70
Gallic Acid Concentrations
Oven Drying of Cooked and Uncooked Ethanolic Extracts of Allium ascalonicum
L. (shallot)
71
SOPHIA MIKAELA M. ABABAO
Block 8 Lot 6, Regency Executive Townhomes
Dasmarinas City, Cavite
+639089872588
sophia.ababao16@gmail.com
I.
II.
III.
IV.
Educational Background
Year/Level
School
Pre-School
Jollikiddie Montessori
Elementary
Philippine Christian University
Junior High School
Rogationist College
Senior High School De La Salle Medical and Health
Sciences Institute
Year
2004-2007
2007-2013
2013-2017
2017-2019
Awards and Honours Received
Awards/Honours
School
Year
Spelling Bee Champion Philippine Christian University 2012-2013
2nd in Diligence (Grade 7)
Rogationist College
2013-2014
rd
3 in Academics (Grade 10)
Rogationist College
2016-2017
Director’s A-List 1st and 2nd De La Salle Medical and Health 2017-2018
Semester
Sciences Institute
Character Reference
Name
Evelyn Segovia
Address
De La Salle Medical and Health
Sciences Institute
Position
Adviser
Profile
Age: 17 years old
Gender: Female
Birthdate: August 16, 2001
Religion: Roman Catholic
Citizenship: Filipino
72
MA. PATRICIA EUNICE T. ANCHETA
Block 6 Lot 37, Springville South II
Molino IV, Bacoor City, Cavite
+639260680334
mapatriciaeunicetancheta@gmail.com
I.
II.
III.
IV.
Educational Background
Year/Level
School
Pre-School
Statefields School, Inc.
Elementary
Statefields School, Inc.
Junior High School
Statefields School, Inc.
Senior High School De La Salle Medical and Health
Sciences Institute
Year
2004-2007
2007-2013
2013-2017
2017-2019
Awards and Honours Received
Awards/Honours
School
Year
Graduated with Honors
Statefields School, Inc.
2016-2017
st
nd
Director’s A-List 1 and 2 De La Salle Medical and Health 2017-2018
Semester
Sciences Institute
Character Reference
Name
Evelyn Segovia
Address
De La Salle Medical and Health
Sciences Institute
Profile
Age: 18 years old
Gender: Female
Birthdate: December 26, 2000
Religion: Roman Catholic
Citizenship: Filipino
Position
Adviser
73
ALLIYAH MARIEL G. ARAÑA
Block 4 Lot 4, Phase 8, Citta Italia
Molino IV, Bacoor City, Cavite
+639750579505
alliyahmariel.arana@yahoo.com
I.
II.
III.
IV.
Educational Background
Year/Level
School
Pre-School
Mother Care Academy Inc.
Pre-School
The Palmridge School
Elementary
The Palmridge School
Junior High School Divine Light Academy
Junior High School Statefields School, Inc.
Junior High School De La Salle University Dasmariñas
Senior High School De La Salle Medical and Health
Sciences Institute
Awards and Honours Received
Awards/Honours
School
Graduated with Honors
De La Salle University –
Dasmariñas
Character Reference
Name
Evelyn Segovia
Address
De La Salle Medical and Health
Sciences Institute
Profile
Age: 18 years old
Birthdate: August 22, 2000
Citizenship: Filipino
Gender: Female
Religion: Roman Catholic
Year
2004-2006
2006-2007
2007-2013
2013-2014
2014-2015
2015-2017
2017-2019
Year
2016-2017
Position
Adviser
74
PAULINE THERESE F. ARRAZ
Block 4 Lot 13, Ville de Soleil, Langkaan I
Dasmariñas City, Cavite
+639476201380
paulinearraz03@gmail.com
I.
II.
III.
Educational Background
Year/Level
School
Pre-School Little House School of Learning
Elementary Immaculate Concepcion Academy of Manila
Elementary Saint Francis Academy – La Salle Supervised
Elementary
Immaculate Concepcion Academy –
South Campus
Junior High School Immaculate Concepcion Academy South Campus
Senior High School De La Salle Medical and Health
Sciences Institute
Awards and Honours Received
Awards/Honours
School
Graduated with Honors Immaculate Concepcion Academy
Director’s A-List 2nd De La Salle Medical and Health
semester
Sciences Institute
Character Reference
Name
Evelyn Segovia
Year
2004-2007
2007-2009
2009-2011
2011-2013
2013-2017
2017-2019
Year
2016-2017
2017-2018
Address
De La Salle Medical and Health
Sciences Institute
IV. Profile
Age: 17 years old
Gender: Female
Birthdate: February 03, 2001
Religion: Roman Catholic
Citizenship: Filipino
Position
Adviser
75
REGINE VIVIENE R. JAVIER
32 A. Monark Subdivision
Pamplona Uno, Las Piñas City
+639565864244
javier.regine@ess.edu.ph
I.
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III.
IV.
Educational Background
Year/Level
Pre-School
Elementary
Junior High School
Senior High School
School
Wishbone Kiddie Camp
Elizabeth Seton School - Main
Elizabeth Seton School – Main
De La Salle Medical and Health
Sciences Institute
Awards and Honours Received
Awards/Honours
School
Graduated with Honors
Wishbone Kiddie Camp
Merit Card holder
Elizabeth Seton School – Main
st
nd
Director’s A-List 1 and 2 De La Salle Medical and Health
Semester
Sciences Institute
Character Reference
Name
Evelyn Segovia
Address
De La Salle Medical and Health
Sciences Institute
Year
2004-2006
2006-2013
2013-2017
2017-2019
Year
2004-2006
2009-2011
2017-2018
Position
Adviser
Profile
Age: 18 years old
Gender: Female
Birthdate: September 2, 2000
Religion: Roman Catholic
Citizenship: Filipino