Feasibility of Phenolic Activity of Guava (Psidium Guavaja) Peel Extract in Delaying Lipid

Oxidation in Frozen Meat Products

In partial fulfillment of the requirements in

UCSP, RaW, Filipino 11, Statistics and Media

We present this research paper

Submitted by:

MERCADO, Orange Allen

JULIAN, Patricia

LACHICA, Myra Venzel

LUGO, Franchesca Nicole

MONSALAUD, Caurrine

ONGNGAD, Althea

Submitted to:

Mr. Mark Erik Menzi

Ms. Abigail Tangalin

Ms. Daphne Raymundo

Mrs. Marcia Viray

Ms. Geraldyn Bambao

March 10, 2017

ABSTRACT
 CHAPTER I

BACKGROUND OF THE STUDY

Meat is a nutritious, protein-rich food which is highly perishable and has a short shelf-life

unless preservation methods are used. Recently, meat preservation became necessary to prevent

contamination by pathogenic organisms or chemicals and losing of optimum qualities of colour,

texture, flavor and nutritive value. Meat preservation makes the food available throughout the

year, establishes prices of food, as there is less scope of shortage of supply to demand, decreases

wastage of food by preventing decay and spoilage of food thus improves the nutrition of the

population thereby decreasing nutritional inadequacies.

In our present time, consumers are increasingly interested about health oriented meat

products. The three sensory properties by which consumers most readily judge meat quality are

appearance, texture and flavor. Appearance of meat products is one of the major determinants of

their appeal to consumers and as a result, sales of the product. (Clydesdale, 1998) The bright,

cherry-red color of meat is an indicator of quality freshness and wholesomeness. Consumers

discriminate against meat cuts that have lost their fresh appearance and meat that becomes

discolored is often ground and marketed in a reduced-value form. Consumers more and more

believe that foods contribute directly to their health (Mollet and Rowland, 2002; Young, 2000)

and are not intended to only satisfy hunger and to provide necessary nutrients but also to prevent

nutrition-related diseases and improve physical and mental well-being of the consumers

(Menrad, 2003; Roberfroid, 2000b).

 Fat, protein, minerals, carbohydrate and water are the constituents of meat (Heinz and

Hautzinger, 2007). The quality of meat and meat products degrade as a result of digestive

enzymes, microbial spoilage and fat oxidation). Lipid oxidation, protein degradation and the loss

of other valuable molecules are the consequence of meat spoilage process. Proteins and lipids

can break down resulting in the production of new compounds causing changes in meat flavor,

tenderness, juiciness, odor and texture. It is therefore, important to understand the causes of

spoilage of meat and meat product in order to develop optimum preservation techniques to

maintain the freshness of these food products (Berkel et al., 2004).

Lipid oxidation, leading to rancidity, is one of the major reasons of meat products' quality

deterioration, mainly because of their increased fat content, the highly comminuted nature of the

raw materials, as well as the possibility of long term frozen storage of the latter before use.

(Frankel 1998) The products of lipid oxidation are also considered as compounds harmful to

human health. Oral intake of fatty acid auto-oxidation products stimulated lipid oxidation in

living organs. Thus, lipid stability is one of the important factors for maintaining meat quality

during storage. Many studies have indicated that lipid oxidation in meat can be effectively

controlled or at least minimized by adding antioxidants.

Cho et al. (1998) ; Jayathilakan et al. (2007a) and McBride et al. (2007) suggested lipid

oxidation in turn deterioration of appearance and microbial growth in meat products can be

controlled, minimized or delayed by using antioxidants. In order to use any antioxidant in food, it

must be safe, easy to incorporate, effective at low concentrations, with no undesirable odor,

flavor or colour, heat stable, nonvolatile and with good carry through properties and cost-

effective (McBride et al. 2007).

 According to Waste & Resources Action Programme (WRAP, 2008), nearly half (46%)

of the avoidable food consumers throw away is in a fresh, raw or minimally processed state, with

an additional 27% thrown away having been cooked or prepared in some way and 20% ready to

consume when purchased. Most of the food consumers throw away (4.1 million tonnes or 61%)

is avoidable and could have been eaten if it had been managed better. In the Philippines, imports

of poultry meat increased by 45 percent from 41 percent in 2008 and it indicates that poultry

meat is part of the processed meat products available in the market. (Ang, 2009).

Antioxidants are chemical compounds that inhibit or retard the lipid oxidation process

and consequently delay the development of rancidity. Hundreds of compounds, both natural and

synthesized have been reported to possess antioxidant properties. Their use in food however is

limited by certain obvious requirements, not the least of which is adequate proof of safety.

Guava (Psidium guajava) is an important tropical fruit, mostly consumed fresh. The fruit

is a berry, which consists of a fleshy pericarp and seed cavity with fleshy pulp and numerous

small seeds. As in many other fruits and vegetables, is also rich in antioxidants that help to

reduce the incidence of degenerative diseases such as arthritis, arteriosclerosis, cancer, heart

disease, and inflammation, and brain dysfunction.

The phenolic compounds are also rich in guava. Miean and Mohamed (2001) claimed

that myricetin, apigenin, anthocyanins and ellagic acid are the examples of phenolic compounds

that contains at high levels in guava. Other polyphenol found in guava in small amount including

quercetin, galic acid, guaijaverin and 3-arabopyranoside guaijaverin. Which turns out guava is a

top among tropical fruits when it comes to disease fighting antioxidants. Consumption of fruits

and vegetables has been associated with reduced risk of chronic diseases such as cardiovascular
disease and cancer. Phytochemicals, especially phenolics, in fruits and vegetables are suggested

to be the major bioactive compounds for the health benefits.

The study will aim to prove that guava peels extract can be effective in delaying the lipid

oxidation in meat. The study also sought to determine the acceptability of frozen meat

incorporated with guava peel extracts.

 Theoretical Framework

This study has the following theories to support the variables of the study: Lipid

Oxidative Theory, Polar Paradox Theory, and Free Radical Theory.

The lipid oxidative theory for atherosclerosis proposes that lipid and/or protein oxidation

products are responsible for lesion formation/development. The lipid oxidative theory suggests

that LDL oxidation contributes to atherogenesis by causing cell injury that leads to arterial

disease. However, a causative link between LDL oxidation and atherogenesis is not firmly

established. It is clear that oxidatively modified LDL particles show chemotactic, cytotoxic and

immunogenic properties that may be exerted prior to interactions in the vascular wall. The

inflammatory and immunogenic nature of atherosclerosis is confirmed by the colocalisation of

macrophages/monocytes and T-lymphocytes in all phases of atherosclerosis, the production of

inflammatory cytokines within intimal thickenings, and the expression of MHC II antigens in the

intima prior to lipid accumulation.

Lipid oxidative theory predicts that appropriate antioxidants will protect against

atherosclerosis. Guava peels has a high quantity of antioxidant such as phenols. Guava also has

high primary antioxidant potential when compared to other local fruits.

Polar paradox theory defines the paradoxical behavior of antioxidants in different media

and justifies the case that polar antioxidants are more effective in less polar media. While

nonpolar antioxidants are more effective in relatively more polar media. It means that polar

antioxidants are more active in bulk lipids than their nonpolar counterparts, whereas nonpolar

antioxidants are more effective in oil-in-water emulsion than their polar homologs. However,

recent results, showing that not all antioxidants behave in a manner proposed by this hypothesis
in oil and emulsion, lead us to revisit the polar paradox and to put forward new concepts,

hypotheses, and theories.

In bulk oil, new evidences have been brought to demonstrate that the crucial site of

oxidation is not the air-oil interface, as postulated by the polar paradox, but association colloids

formed with traces of water and surface active molecules such as phospholipids. The role of

these association colloids on lipid oxidation and its inhibition by antioxidant is also addressed as

well as the complex influence of the hydrophobicity on the ability of antioxidants to protect

lipids from oxidation. In oil-in water emulsion, we have covered the recently discovered non-

linear (or cut-off) influence of the hydrophobicity on antioxidant capacity.

Free radical theory can be defined as reactive chemical species having a single unpaired

electron in an outer orbit. Free radicals induce oxidative stress, which is balanced by the bodys

endogenous antioxidant systems with an input from co-factors, and by the ingestion of

exogenous antioxidants. If the generation of free radicals exceeds the protective effects of

antioxidants, and some co-factors, this can cause oxidative damage which accumulates during

the life cycle, and has been implicated in aging, and age dependent diseases such as

cardiovascular disease, cancer, neurodegenerative disorders, and other chronic conditions.

Oxidative damage to proteins, lipids, and DNA accumulates and increases with age, and

is associated with age-related diseases which can be compared to the shelf life of the meat

product. The meats are being damaged through the lipid oxidation. Using antioxidants such as

guava can help delay the process of aging meat.

 Review of Related Literature

Foreign Literature

Guava (Psidium guajava) is an exotic fruit member of the fruit family Myrtacea. Guava,

goiaba or guayaba are some of the names given to the apple of the tropics, popular for its

penetrating aroma and flavor. The guava has been cultivated and distributed by man and animals

for so long that its place of origin is uncertain, but it is believed to be an area extending from

southern Mexico to Central America. It was soon adopted as a crop in Asia and in warm parts of

Africa. Currently, its cultivation has been extended to many tropical and subtropical parts of the

world, where it also thrives well in the wild. In India, guava cultivation has been estimated at

125,327 acres yielding 27,319 tons annually. It is common found in warm climates area because

it can survive only a few degrees of frost.

The peel and seed fractions of some fruits possess higher antioxidant activity than the

pulp fractions. Guo et al (2003) states that most of fruit peel and seed fractions were stronger

than the pulp fractions in antioxidant activity based on their FRAP values. The contribution of

vitamin C to the FRAP value of fruit pulps varied greatly from fruit to fruit as calculated.

Therefore, the peel and seed fractions of fruits may potentially contain more antioxidants

quantitatively or qualitatively than the pulp fractions. Besides, Nurliyana et al., (2010) stated that

previous studies have determined the antioxidant levels and activities in tropical fruits such as

mangosteen, papaya and star fruit, covering various parts of fruits. However, investigations on

the antioxidant activities for fruit peels and seeds are lacking due to their low popularity and

commercial application (Soong and Barlow, 2004). According to Caro and Piga (2007), seeds

and peels of Italian fresh fig fruits cultivars showed higher antioxidant capacity and phenolic
content than the edible portions. That is why, guava, like other tropical fruits, is believed to be

rich in antioxidants but information specifically on antioxidant levels and properties in its peels

are still lacking.

A growing demand in developed countries for safer foods and cleaner production

processes has been observed in recent times. The processing of agro product results the

formation of waste materials in high amount (Martin et al. 2012) and commonly are in the form

of peels, seeds and oilseed meals (Stanikova et al. 2005) and accumulation of the wastes pose a

problem to the environment as they are prone to microbial spoilage (Garcia et al. 2005). In

addition, the cost of drying and storage or transportation concern with financial limitation for

waste utilization. All such factors and lack of proper management and non-availability of

technology, these wastes are either disposed of for decomposition and compost production or

incinerated (Ajila et al. 2010). This has led to a drive, by the agroindustry, for waste reduction

and upgrading techniques to reduce costs and achieve new sources of income. In fact, agro waste

may be a source of high-added value products potentially useful as beneficial food constituents,

food flavors and antioxidants or as cosmetics, chemo preventive agents or drugs.

According to Shodhganga (2013), antioxidant compounds in natural products play an

important role as a health protecting factor. Scientific evidence suggests that antioxidants reduce

the risk for chronic diseases. Most of the antioxidant compounds are derived from plant sources

and belong to various classes of compounds with a wide variety of physical and chemical

properties. The main characteristic of an antioxidant is its ability to trap free radicals. Highly

reactive free radicals and oxygen species are present from a wide variety of sources. Antioxidant

compounds like phenolic acids, polyphenols and flavonoids scavenge free radicals such as
peroxide, hydroperoxide or lipid peroxyl and thus inhibit the oxidative mechanisms in nucleic

acids, proteins, lipids or DNA and can initiate degenerative disease.

Due to the chemical diversity of phenolic compounds and the complexity of composition

in plant samples, it is difficult and inexpensive to separate each phenolic antioxidant and study it

individually. Moreover, study of total antioxidant power of a complex sample is often more

valuable because of the cooperative action of antioxidants. Therefore, it is desirable to

standardize convenient screening methods for quick quantification of antioxidant property of

phenolics in natural products. A variety of antioxidant assays such as ferric ion reducing

antioxidant power (FRAP) has been widely used for quantification of antioxidant capacity of

phenolic samples from fruits and vegetables. The Folin-Ciocalteu antioxidant capacity assay (F-

C assay, or total phenolics assay) is also considered as another antioxidant capacity assay

because its basic mechanism is as oxidation/reduction reaction though it is used as a

measurement of total phenolics content (Huang et al. 2005).

When the body's natural defenses fail to protect against oxidative stress, balance may be

restored by the consumption of nutraceuticals that have antioxidant properties. According to

Wildman (2001), nutraceuticals include isoprenoid derivatives (i.e.carotenoids and tocopherols);

phenolic compounds (tannins, anthocyanins, flavonoids); amino acid-based (isothiocyanates,

folate, choline); carbohydrates (oligosaccharides,ascorbic acid); fatty acids (polyunsaturated fatty

acids, lecithin, and conjugated linoleicacid); minerals (Ca, K, Zn); and microbiols (pre- and pro-

biotics). In particular, the phenolic compounds have been shown to exert potent antioxidant

capacities, particularly the flavonoids, (Wildman, 2001; Biesalski, 2001) to protect against many

diseases (Ishige et al., 2001; Casaburi et al., 2013). As secondary metabolites in plants, phenolic

compounds are not directly involved with respiration, photosynthesis, or nutrient assimilation
(Blumberg and Block, 1994) but rather play important roles in protecting a plant from insects,

disease, and herbivores. Phenols also attract pollinators and thus help to ensure the propagation

of the next generation in plant to plant competition (Wildman, 2001).

From the book Lipid Oxidation, the kinetic stability of a radical is largely controlled by

steric factors. When the radical center is crowded, the radical becomes less reactive and can have

a longer life-time. Aromatic com-pounds that can form allylic radicals show similar benzylic

stabilization. It is also said that, if the radical center is sterically crowded by bulky tertiary butyl

substituents, the allylic radical intermediates formed by hydrogen transfer have kinetic stability

that imparts important antioxidant properties. Hence, when phenolic compounds contain three

bulky tertiary butyl substituents, they form persistent radicals after hydrogen donation and inhibit

lipid oxidation by interrupting the propagation of free radicals. Rancidity of food is a major

problem in meat industry, and it is because of increasing emphasis on the use of polyunsaturated

fish oils and vegetables, discontinuing the use of synthetic antioxidants, and fortification of

cereals in adults and infant foods with iron. The oxidation in lipids does not only produces

undesirable rancid flavors in foods but can also decrease the nutritional quality. Effective control

methods against lipid oxidation include the use of metal inactivators (or chelators), minimizing

the loss of natural tocopherols and exposure to air and light during processing, hydrogenation

and the use of antioxidants.

Foreign Study

Phenols are present in all plant based systems with approximately 8000 different types

categorized by their structure (Pescarmona, 2007), which in all categories include the aglycones
(phenols that are not bonded to another molecule) or the conjugated phenols (e.g., glycosylated

or esterified with a single or multiple sugar units or other phenols, etc.).

Phenols protect against oxidation in both plants and humans by various mechanisms.

First, a phenol is able to transfer its H-atom to a radical directly, which is termed the H-atom

transfer. Second, phenols transfer an electron to deactivate a free radical followed by proton.

Third, phenols transfer an electron to deactivate a free radical followed by proton abstraction via

proton-coupled electron transfer. Fourth, the sequential proton- loss electron transfer can occur

where deprotonation of phenol is followed by electron transfer from its phenoxyl anion to a free

radical (Klein and Lukes, 2006). Phenolic compounds become resonance- stabilized radicals

after donation making the free radical stable (Shahidi et al., 1992) (Figure 4). The quantity and

the type of phenols present control the efficiency of phenolic rich natural systems to act as potent

free radical scavengers (Musialik et al., 2009).

FRAP assay is commonly used to study the antioxidant capacity of plant materials. The

antioxidant capacity of fruits extracts is determined by the ability of the antioxidants in these

extracts to reduce ferric iron to ferrous in FRAP reagent, which consists of2, 4, 6-tris (1-pyridyl)-

5-triazine (TPTZ) prepared in sodium acetate buffer, pH 3.6. The reduction of ferric iron in

FRAP reagent will result in the formation of a blue product (ferrous TPTZ complex) whose

absorbance can be read at 593 nm (Alothman et al., 2009). Guo et al., (2003) states that there are

many different antioxidants contained in fruits and it is very difficult to measure each antioxidant

component separately. Therefore, several methods have been developed to evaluate the total

antioxidant activity of fruits or other plants and animal tissues. Among them, Trolox equivalent

antioxidant capacity, total radical absorption potentials, oxygen radical absorption capacity

assays are the representative methods frequently used in various investigations. However, none
of the methods mentioned above can be treated as a total antioxidant capacity assay because

what they really measure is the capacity of antioxidants in scavenging specific radicals,

inhibiting lipid peroxidation or chelating metal ions. The commonly used method for

determining antioxidant activity is FRAP assay. This is based on several reasons that were first,

the FRAP assay treats the antioxidants in the samples as reductants in a redox-linked

colorimetric reaction. Second, the procedure of FRAP assay is relatively simple and easy to be

standardized. One possible disadvantage with FRAP assay is the fact that this assay does not

react fast with some antioxidants, such as glutathione. However, we consider that FRAP assay is

still suitable for assessment of antioxidant activity of fruit samples because only limited amounts

of plant glutathione are absorbed by humans. (Guo et al., 2003)

Thaipong, 2006 affirms in his study that the FRAP technique showed high

reproducibility, was simple, rapidly performed and showed the highest correlation with both

ascorbic acid and total phenolics compared to the other methods. Therefore, it would be an

appropriate technique for determining antioxidant in guava fruit extract.

In the study entitled Effects of Mango (Mangifera indica L.) Extract on Frozen Chicken

Meat Balls Storage Quality the researchers determined the FRAP according to Oyaizu (1986).

Different concentrations of mango (1.6-9.6 mg) in 1.0ml of distilled water were mixed with 2.5

ml phosphate buffer (0.2 M, pH 6.6) and 2.5 ml potassium ferricyanide [K3Fe(CN)6] (1%). The

mixtures were incubated at 50C for 20 min. Aliquots (2.5 ml) of trichloroacetic acid (10%) were

added to the mixtures, which were then centrifuged for 10 min at 3000 rpm. The supernatant (2.5

ml) was mixed with 2.5 ml distilled water and 0.5 ml of 1% FeCl3. The absorbance of the

reaction mixtures was measured at 700 nm. Increased absorbance of the reaction mixture

indicates increased reducing power. While the total phenolic content were determined through
the help of Waterman and Mole (1994) using tannic acid as a standard phenolic compound.

Briefly 0.05 mg extract was diluted with 10 ml of distilled water in a volumetric flask. Then 1.0

ml of solution was taken out into a volumetric flask and 0.5 ml of Folin-Ciocalteu reagent was

added. The solution was stirred so that the content of the flask mixed thoroughly. Three minutes

later, 1.0 ml Na2CO3 (2%) was added and the solution marked up to 10 ml with distilled water.

The mixture was then stored for 1-hr in dark condition. The absorbance of the react mixture was

measured at 760. Lastly, the peroxide value was determined based on PORIM (1995) method.

1.0 ml of fat is weighed in 250 ml volumetric flask. 30 ml of acetic acid-chloroform (1:3) was

added and the flask was swirled until the sample was dissolved in the solution. Saturated

potassium iodide (0.5 ml) was added and the flask was swirled again before the addition of 30 ml

of distilled water. A few drops of starch solution were added as the indicator and the mixture was

titrated with 0.01N sodium thiosulphate until the blue color disappears.

According to the study entitled Antioxidant Properties of Guava Fruit: Comparison with

Some Local Fruits, ss the fruit ripens, TPC decreases but AAC (Ascorbic acid content)

increases. According to Taylor (1993), the decrease in polyphenol content of guava fruit causes a

loss in astringency during ripening of the fruit. The increase in AAC as the fruit matures is due to

the breakdown of starch to glucose which is used in the biosynthesis of ascorbic acid. The DPPH

scavenging activity of the unripe guava as measured by the AEAC (ascorbic acid equivalent

antioxidant capacity) value is primarily due to its higher TPC relative to AAC. Decrease of

AEAC during ripening suggests that the antioxidant activities of guava fruit declined during fruit

ripening. Increase in AAC was observed during storage of ripe and unripe guava. The trend for

TPC appeared to increase in unripe samples but decrease in ripe samples. However due to the
small changes observed and the limited number of samples, a definite conclusion on TPC trend

during storage cannot be reached and more studies are required.

Local Literature

With the high demand for chicken meat, stemming from growing household

consumption, a rapid expansion of institutional buyers like fast-food chain owners as well as its

potential prospect as an export product, it is no surprise that the poultry sector has showed strong

growth (in value and volume) over the last 14 years (2000-2013). Poultry has eroded shares of

other meats with this trend evident also in the global meat market.

Poultry is mostly defined by chicken production, generating both meat and eggs. Chicken

meat is the most utilized species of poultry, mainly for food production. In the Philippines,

chicken ranks first in terms of economic importance as source of meat and eggs
 Assumption of the Study

The researchers assumptions are as follows:

1. Guava is a tropical fruit. It is an ever-green fruit rich in high-profile nutrients. The health

benefits of guava include the treatment of diarrhea, diabetes, concentration, cancer

prevention, cough and colds.

2. Meat is an animal flesh that is eaten as food.

3. Lipid Oxidation is an oxidative deterioration of lipids containing any number of carbon

double bonds (fatty acids, cholesterol). It is also a major cause of deterioration in the

quality of meat and meat products.

 Definition of Terms

The following terms are operationally and conceptually defined as utilized in the study

for letter understanding and clarity of the direction of the study.

Delay/Delaying. It is a process that is expected to affect the lipid oxidation in frozen

meat products. It means to keep the food for a longer time.

Ferric Reducing Antioxidant Power (FRAP). It is an automated test measuring the

ferric reducing ability of plasma.

Frozen Meat Products. These are the products that will be used in the study i.e. chicken

breasts. It is commonly found in farm animals like chicken, cows, or pigs.

Guava (Psidium guajava) Peel Extract. The independent variable in the study. It is the

fruit that will be used to retard the lipid oxidation of meat products.

Lipid oxidation. It is the dependent variable in the study. It is a process when the meat

lose its properties and nutritional values.

Peroxide Value. It is used to assess the concentration of peroxide in oil and to know the

extent of spoilage has occurred.

Total Phenolic Content. It is the total measure of the phenols in the guava.

Microbial Growth. It is the process of increase in the number of cells, cell activity, and

cell mass of bacteria.

 Statement of the Problem

This study will determine the effectiveness of guava peel extract in delaying lipid

oxidation in frozen meat products.

Specifically, it will aim to respond to the following questions:

1. Is guava peel extract effective in delaying the lipid oxidation in frozen meat products in terms

of:

1.1 Total Phenolic Content;

1.2 Ferric Reducing Antioxidant Power; and,

1.3 Peroxide Value?

2. Is there a significant difference between the experimental groups in terms of:

2.1 Total Phenolic Content;

2.2 Ferric Reducing Antioxidant Power; and,

2.3 Peroxide Value?

3. Is there a significant difference between the experimental group and controlled group in terms

of:

3.1 Total Phenolic Content;

3.2 Ferric Reducing Antioxidant Power; and,

3.3 Peroxide Value?

4. What is the implication of the study towards the delaying lipid oxidation in frozen meat

products?

5. Is the study beneficial and practical to the community and environment?

 Hypothesis

The hypotheses will be tested for further analysis of data to answer the major problem:

1. The guava peel extract is not effective in delaying the lipid oxidation in frozen meat

products in terms of total phenolic content, ferric reducing antioxidant power, and

peroxide value

2. There is no significant difference between the experimental groups in terms of total

phenolic content, ferric reducing antioxidant power, and peroxide value

3. There is no significant difference between the experimental group and controlled group in

terms of total phenolic content, ferric reducing antioxidant power, and peroxide value

4. There is no effect in prolonging the shelf life of the frozen meat products.
 Scope and Limitation

The study will evaluate the phenolic activity of guava peel extract in delaying lipid

oxidation in frozen meat products. The researchers will use chicken breasts in conducting the

experiment, which will be bought in the Baguio City Public Market. The guavas will be bought

also in the Baguio City Public Market. The peels will be the only part to be extracted. Using

Complete Randomized Design, there will be three treatments and three replications per

treatment: Treatment A (50 grams guava peel extract), Treatment B (100 grams guava peel

extract), and Treatment C (no guava peel extract). The meats for the lipid oxidation test will be

stored in the refrigerator for 30 days, giving the treatments and performing lipid oxidation for 10

days. The samples will undergo phytochemical screening at the Department of Science and

Technology Cordillera Administrative Region (DOST-CAR). The TPC, FRAP, and Peroxide

value will be tested in research institute in DOST-CAR as well. The experimentation will be

conducted from the period of March to April 2017.

The study is limited in using only the peels of unripe guavas. The researchers will only

test the total phenolic content, ferric reducing antioxidant power, and peroxide value of the meat.
 Significance of the Study

This study will be beneficial in evaluating the phenolic activity of guava peels to the

following:

Community. This will be helpful to the community in a way that they could delay lipid

oxidation in meat by giving them the proper knowledge and methods about this process. In

addition to certain food storage and deterioration concerns in the community, in meat products, it

may involve various technological improvements influencing the final quality of the products.

Lipid oxidation must be highlighted due to the modification of both the sensory characteristics

and the shelf-life of meat products. It will also be helpful to the community particularly for the

increasing rate of household and institutional consumption in fast-food and small-time

businesses. Lastly, the study can also be beneficial for exportation of meat products by

increasing the period of its shelf-life and quality during the transportation.

Health. This study will be beneficial for helping in reducing the impact of chemical changes

and promote oxidative stability; with the use of natural antioxidants which has gained ground

owing to the health and safety advantages linked to its effectiveness at reducing or delaying lipid

oxidation. This study will also be helpful to prevent foodborne and spoilage bacteria in fresh,

meat products to decrease the chances of acquiring certain types of diseases.

Education. This study will help future students to acquire new knowledge in the field of

nutritional and agricultural sciences. It can provide them information about the antioxidants

produced, to describe the use of different compounds to enhance lipid oxidative stability.

Environment. This study will help lessen the rotten meats in the dumpster and can help diminish

animals to be slaughtered. It will also lessen the emission of carbon dioxide, methane (from
anaerobic decomposition) , amines (from the decomposition of protein), and hydrogen sulphide

(again, from protein decomposition) which are contributors to pollution in the atmosphere due to

the decaying food reaction.

Research. This study will benefit future researchers for this can contribute new knowledge and

idea to others to help improve the said industry. Furthermore, this study can help researchers

acquire ideas on further delay of lipid oxidation to frozen meat products.

 CHAPTER II

PARADIGM OF THE STUDY

This chapter contains the concepts regarding the methodology of the study which is

divided into parts: the research design, research instrument, subjects of the study sampling

technique, sampling procedure, research procedure, and statistical treatment used in the study. It

also presented the strategies for instituting the credibility of the data and the ethical consideration

to be scrutinized in the conduct of the research.

 Conceptual Framework

Input Process Output

Extraction through
blender
Gravity Filtration
Chicken breasts
Complete
Randomized Design
Three Treatments
Guava Effective
Three Replications
Peel Refrigerated storage in
Total Phenolic
Extract Content Delaying
Ferric Reducing
Lipid Oxidation
Antioxidant Power
Peroxide value
ANOVA
Paired t-test

Figure 1. Paradigm of the Study

Phenolic Activity of Guava (Psidium Guajava) Peel Extract in Delaying Lipid Oxidation in
Frozen Meat Products
 Conceptual Paradigm

Figure 1 shows the conceptual framework of the study Phenolic Activity of Guava Peel

Extract (Psidium Guajava) in Delaying Lipid Oxidation in Frozen Meat Products. The guava

peel extract, which is the input and independent variable, undergo the process of extraction

through blender and gravity filtration method. Using Complete Randomized Design, the study

will have three (3) treatments: Treatment A (50 grams guava peels extract), Treatment B (100

grams guava peels extract), and the Treatment C (controlled group), with three (3) replications

each. The treatments will undergo three (3) testings in terms of total phenolic content, ferric

reducing antioxidant power, and peroxide value. Data and results will undergo an Analysis of

Variance (ANOVA) and the paired t-test as its statistical hypothesis testing. Thus, the output of

the study is effective in delaying lipid oxidation.

 Research Design

The researchers will use the complete randomized design (CRD). In complete

randomized design type of research, the researchers will randomly divide the experimental units

and randomly assigned to the treatment to see the outcome of the treatment and whether the

desired outcome is achieved. In order to discuss the problem, background information and

previous studies done will be collected, and the hypothesis, which answers the problem, will be

drawn. In order to prove the precision of the hypothesis, empirical data will be collected via

observation. All procedures will be duly recorded.

 Research Instrument

The testing that will be used in this study are the total phenolic content, ferric reducing

antioxidant power, and peroxide value in order to gather results from the study.

A simple, automated test measuring the ferric reducing ability of plasma, the FRAP

assay, is presented as a novel method for assessing antioxidant power. Ferric to ferrous ion

reduction at low pH causes a colored ferrous-tripyridyltriazine complex to form. FRAP values

are obtained by comparing the absorbance change at 593 nm in test reaction mixtures with those

containing ferrous ions in known concentration.

The peroxide value is a parameter specifying the content of oxygen as peroxide,

especially hydro peroxides in a substance. The peroxide value is a measure of the oxidation

present. Peroxides and similar products which oxidize potassium iodide under the conditions of

the test will contribute to the peroxide value. Variations in procedure may affect the results.

Peroxide values are expressed either in milliequivalents of peroxide/kg or millimoles of

peroxide/l.

The Guava peels extract will be crushed using a blender. It will be weighed by using

calibrated weighing scale and the water is measured by a measuring cup. The researchers will

use an injection to insert the extract in the chicken breasts.

 Subjects of the Study

The experimentation will be conducted at St. Louis School, Inc. Center High School

Department Science Laboratory. Therefore the researchers will buy chicken breast in order to

gather results. The researchers need nine (9) chicken breasts.

 Sampling Technique

The study used a Purposive Sampling, wherein the sampling is under Non-probability

Sampling. In the said technique, wherein a researcher chooses specific sample within the

population to use for a particular study. Unlike random studies, which deliberately include a

diverse cross section of ages, backgrounds and cultures, the idea behind purposive sampling is to

concentrate on samples with particular characteristics who will better be able to assist with the

relevant research.
 Sampling Procedure

There are three steps in selecting a sample using purposive sampling:

1. The researchers will go to the Baguio City Public Market to search for chicken breast.

2. The chicken breast must be fresh before the experimentation.

3. The researchers will assign three chicken breasts in each treatment.

There are also steps in acquiring a sample using simple random sampling in

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 Research Procedure

A. Materials

1. To carry out the experiment, the following will be used in the study: Nine (9) chicken

breasts, four (4) 10cc syringes, and refrigerator. These are for the main experimentation.

Members of the group will be assigned to buy the chicken breasts. The syringes will be

gathered on the laboratory.

2. In extracting the guava peel, the following materials and instruments that will be used

are: eight (8) unripe guava, knives, two (2) reagent bottles, one (1) blender, one (1) iron

stand, one (1) iron ring, one (1) funnel, one (1) 500 mL beaker, two (2) filter paper, and

distilled water. The materials will be borrowed by the team in the St. Joseph College-

High School Department Chemical Laboratory. The guava will be brought by members

of the group.

3. Observation will be conducted and results of the experiment will be tallied, analyzed

and interpreted.

B. General Procedure

1. Extraction of Unripe Guava Peels

a) The materials will be borrowed in the laboratory and the guava will be

gathered and brought by the team members.

b) The researchers will peel the unripe guava to make it easier to extract.
 c) The guava peels will be chopped for a finer bit of the peel.

d) The researchers will blenderize the gathered bits of peel using a blender until

all peels are fine.

e) The blenderized peels will be poured into the gravity filtration set-up.

e) The extract will be sealed into two reagent bottles with different measurements

of the extract: 50 grams and 100 grams.

f) The bottles will be then stored in a refrigerator for preservation.

2. Evaluation of the phenolic activity of guava peel extract in delaying lipid oxidation in

frozen meat products.

a) The chicken breasts will be bought on the new market at Olongapo City.

b) For the pretest, the researchers, will test the total phenolic content (TPC) using

gallic acid as the standard phenolic compound, ferric reducing antioxidant power

(FRAP) using Oyaizus method, and peroxide value (PV) by using the PORIM

method of the nine (9) chicken breasts by cutting a small portion of the meat.

c) The results will be gathered by the team members.

d) The chicken breasts will be stored for 30 days

e) Every 10 days, the chicken breasts will be tested for the TPC, FRAP, and PV

and afterwards the chicken breasts will be smeared by the extract using the 10cc

syringe.

f) Results will be observed and data will be gathered, statistically tallied,

analyzed, and interpreted. The Analysis of Variance (ANOVA) and paired t-test

will be used to determine whether there is a significant difference between the

experimental groups and controlled groups in terms of phenolic compound, ferric

reducing power, antioxidant activity, and peroxide value.