CHAPTER I

THE PROBLEM AND REVIEW OF RELATED LITERATURE

Introduction

Our bodies are battlegrounds against infection and diseases.

Normal body functions, such as breathing or physical activity, and

other lifestyle habits (such as smoking) produce substances called

free radicals that attack healthy cells. When these healthy cells

are weakened, they are more susceptible to cardiovascular disease

and certain types of cancers. Antioxidants — such as vitamins C

and E and carotenoids, which include beta-carotene, lycopene and

lutein — help protect healthy cells from damage caused by free

radicals.

The human body naturally produces free radicals and the

antioxidants to counteract their damaging effects. Free radicals

might occur either by the accidents of chemistry or due to specific

metabolic purpose in the body. The free radicals produced by

either way have different reactivity with some leading to damage

to biomolecules such as DNA, lipids and proteins. However, in most

cases, free radicals far outnumber the naturally occurring

antioxidants. In order to maintain the balance, a continual supply

of external sources of antioxidants is necessary in order to obtain

the maximum benefits of antioxidants. Antioxidants benefit the

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body by neutralizing and removing the free radicals from the

bloodstream. Antioxidants can react with free radicals during the

oxidation process by acting as a reactive species scavenger and

liberating catalysts, so antioxidants can be used to reduce the

oxidative process.

In addition, Goyal and Brahma (2014) said that in the process

of economic development, with the increase in income, human society

tends to care more about their health. Therefore, demand for

healthy herbal organic foods developed from various plants has

also increased. Production of more efficient and productive food

items by the researchers are on demand.

One such plant with multiple qualities is bamboo. Bamboo has

been used over centuries by the humans both in daily life and for

medicinal purpose in China and other Asian countries. The earliest

scientific evidence of use of bamboo in traditional medicine dates

back to 1963. This marked the beginning of the use of bamboo as

medicine which was followed by series of research carried out by

different workers since then. Bamboo has attracted attention world

over due to its high antioxidant content and therapeutic effects

on inflammation, fatigue, cancer, hyperlipidemia, diabetes, aging

and hypertension.

A study by Gong et al., (2016) explored that the wide range

of antioxidant activities of bamboo extracts. His study indicated

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the potential of bamboo shavings as a source of natural

antioxidants or nutraceuticals with potential application in

reducing oxidative stress with consequent health benefits.

The research of, Sangeetha et al. (2015) aimed to provide a

safe and effective test drug that can eradicate the chain reaction

of free radicals, specifically the DPPH (2,2-diphenyl-1-

picrylhydrazyl) which contains a dark-colored crystalline powder

composed of stable free-radical molecules. The main of the test

drug were the leaves of Schizostachyum brachycladum (Golden Buho).

The bamboo is a less explored plant with high therapeutic

potential. With the presence of several species of this plant in

the Philippines, the researchers are interested to study its other

uses, aside from its use in food and craft making.

Review of the Related Literature

This part deals with the literature and studies related to

the problem under investigation. Related literature and studies

were included not only to enrich background for the development of

the study but also to support the discussions, analysis, and

interpretation of the findings.

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A. Conceptual Literature

Botanical Description of Golden Buho (Schizostachyum brachycladum)

Schizostachyum brachycladum is an

evergreen bamboo, forming dense clumps of

culms 10 - 15 metres tall from short, woody

rhizomes. The thin-walled, woody culms can

Figure 1: Golden
be erect or arching; they are 4-8cm in
Buho
diameter with internodes 25 - 50cm or longer.

This species is one of the economically important bamboos in

the Philippines, providing material for a wide range of uses. It

grows extensively in large, natural stands but there are no large-

scale plantations. Its exploitation is generally unregulated and

no economic or production statistics are available. Consumption

and trade are mainly local in rural areas, whilst in the northern

Philippines a processing plant has been established to make ply

bamboo from natural stands. Usually harvested from large, natural

stands, the plant is also cultivated occasionally

It is a densely tufted, sympodial bamboo. The culm is erect

to ascending, 10-15 m tall, 4-8 cm in diameter, wall 4-5(-10) mm

thick. Internodes are 25-50(-80) cm long, glabrous, and green. The

nodes are oblique. Branches are several to too numerous at the

upper nodes. Culm sheath 24-26 cm long, up to 33 cm wide at the

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base, persistent, covered with yellowish, sharp hairs. Blade is

lanceolate, 9 cm × 1.9 cm, reflexed, shortly pubescent on both

surfaces, hairs mostly deciduous. Ligule very short, minutely

ciliate; auricles not distinct. Flowering branches arise at the

upper nodes, bearing groups of pseudospikelets at their nodes.

Spikelet are linear-lanceolate, about 15 mm × 1.5 mm, glabrous,

comprising 2 empty glumes and one fertile floret. Caryopsis is

oblongoid, 6-8 mm × 1-1.5 mm, brown.

Young shoots emerge during the rainy season and develop to

their full height in 4-6 months. Culms become mature in 1-2 years.

They reach their maximum diameter at 5 m height. A healthy clump

produces several young shoots annually, up to about 10% of the

number of mature culms. The number of culms per hectares in natural

stands averages about 9000, but can be as high as 25 000 in dense

stands. The dry weight rate of the above ground parts of a culm is

approximately 89% for the culm, 7% for the branches, 4% for the

leaves. In the Philippines, flowering is from January to May,

fruiting from June to July.

Origins and Geographic Distribution

Schizostachyum brachycladum is native to the Philippines and

occurs extensively in the Provinces of Pangasinan, La Union, Ilocos

Norte, Ilocos Sur, Leyte and on the Islands of Panay and Basilan.

It is cultivated occasionally.

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 Grows extensively in large, natural stands throughout the

country in thickets and secondary forests, sometimes nearly

exclusive occupying large areas at elevations up to 1,500 meters

above sea level.

Taxonomic Classification of Golden Buho (Schizostachyum

brachycladum)

Kingdom : Plantae

Phylum/Division : Tracheophyta

Class : Liliopsida

Order : Poales

Family : Poaceae

Genus : Schizostachyum

Species : Schizostachyum Brachycladum

Scientific Name : Schizostachyum Brachycladum

Common Name : Golden Buho

About DPPH (2,2-diphenyl-1-picrylhydrazyl)

DPPH is a common abbreviation for the organic chemical

compound 2,2-diphenyl-1-picrylhydrazyl. It is a dark-colored

crystalline powder composed of stable free-radicalmolecules. DPPH

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has two major applications, both in laboratory research: one is a

monitor of chemical reactions involving radicals, most notably it

is a common antioxidant assay, and another is a standard of the

position and intensity of electron paramagnetic resonancesignals.

DPPH is a free radical which shows hydrogen acceptor ability

towards antioxidants. It is commonly used in DPPH assay for

measuring the antioxidant activity of different natural samples

such as wine, fruits, herbal tea, etc.

DPPH is a well-known radical and a trap ("scavenger") for

other radicals. Therefore, rate reduction of a chemical reaction

upon addition of DPPH is used as an indicator of the radical nature

of that reaction. Because of a strong absorption band centered at

about 520 nm, the DPPH radical has a deep violet color in solution,

and it becomes colorless or pale yellow when neutralized. This

property allows visual monitoring of the reaction, and the number

of initial radicals can be counted from the change in the optical

absorption at 520 nm or in the EPR signal of the DPPH.

As a stable and well-characterized solid radical source, DPPH

is the traditional and perhaps the most popular standard of the

position (g-marker) and intensity of electron paramagnetic

resonance (EPR) signals – the number of radicals for a freshly

prepared sample can be determined by weighing and the EPR splitting

factor for DPPH is calibrated at g = 2.0036. DPPH signal is

7
convenient by that it is normally concentrated in a single line,

whose intensity increases linearly with the square root of

microwave power in the wider power range. The dilute nature of the

DPPH radicals (one unpaired spin per 41 atoms) results in a

relatively small linewidth (1.5–4.7 Gauss). The linewidth may

however increase if solvent molecules remain in the crystal and if

measurements are performed with a high-frequency EPR setup (~200

GHz), where the slight g-anisotropy of DPPH becomes detectable.

Whereas DPPH is normally a paramagnetic solid, it transforms

into an antiferromagneticstate upon cooling to very low

temperatures of the order 0.3 K. This phenomenon was first reported

by Alexander Prokhorov in 1963.

Antioxidants

Antioxidants are man-made or natural substances that may

prevent or delay some types of cell damage.

Antioxidants come up frequently in discussions about good

health and preventing diseases. These powerful substances, which

mostly come from the fresh fruits and vegetables we eat, prohibit

(and in some cases even prevent), the oxidation of other molecules

in the body. The benefits of antioxidants are very important to

good health, because if free radicals are left unchallenged, they

can cause a wide range of illnesses and chronic diseases. Examples

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of antioxidants include: Beta-carotene, Lutein, Lycopene,

Selenium, Vitamin A, Vitamin C, Vitamin E.

Tripathi et al. (2015) mentioned that excessive free radical

production generally leads to oxidative stress which is

responsible for the onset and progression of several diseases.

Natural anti-oxidants have the ability to protect organisms from

damage caused by free radical-induced oxidative stress and

therefore can be used to fight a variety of ailments and diseases.

Bambusa vulgaris has been traditionally used in folkloric medicine

and most of the medicinal properties have been attributed to its

antioxidant activities. Since, this study revealed that Bambusa

nutans has higher polyphenolic content and antioxidant activity

than Bambusa vulgaris. Therefore, it can be speculated that Bambusa

nutans can also be potentially useful as a natural source of

antioxidant or in medicine.

Mere large doses of diet-derived antibody were thought to be

important to stay healthier for long time, but with the passage of

time and development of science and technology the supply of “pro-

oxidants‟ is thought to be a better option. Bioactive compounds

like ascorbic acid, carotenoids, tocochromanols and phenols are

antioxidants. The bamboo leaf extract (BLE) is thought to be good

source of natural antioxidants and also have great pharmaceutical

potential. BLE is mainly composed of flavonoids, lactones and

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phenolic acid. The flavonoids are represented mainly by the

flavones C- glycosides which include homoorientin, isovitexin,

orientin and vitexin. Apart from this quercetin, luteolin, rutin,

caffeic acid, p-coumaric acid, chlorogenic acid and tricin are

also present. The flavonoid content was recorded to be 3.44% in

different bamboo leaves species.

Commercial Antioxidants

L-Ascorbic Acid

Pure Vitamin C is known as L-Ascorbic Acid. The “L” in front

of Ascorbic is a reference to how the molecule itself rotates to

light and refers to its source. L-Ascorbic Acid comes from natural

sources such as oranges.

L-Ascorbic Acid found in topical skincare products carries a

concentration between 5 percent to 15 percent. A higher

concentration can irritate the skin. L-Ascorbic Acid with a

concentration between 5 percent to 15 percent usually has a lower

pH, making it a serum that's unsuitable for extremely sensitive

skin types.

L-Ascorbic Acid products penetrate the skin tissue and are

more active in collagen production. This, in turn, increases

firmness in the skin. L-Ascorbic Acid is a rejuvenating vitamin

ingredient that also plays a part in the reduction of photo-damage

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caused by harmful UVB rays. Because L-Ascorbic Acid plays a major

role in anti-aging treatments, skin firmness, overall repair and

brightening, it also serves to rejuvenate the skin.

Conditions such as aging skin, hyperpigmentation and post-

inflammation hyperpigmentation benefit greatly from L-Ascorbic

Acid skin care products. People with rosacea and sensitive skin

conditions should avoid this Vitamin C ingredient as it may cause

irritation. Oily, dry and combination skin types can benefit

greatly from L-Ascorbic Acid products.

Other Uses of Golden Buho

Bamboo has been used for eons for many applications, from a

food source to a building material. But with the age of modern

materials, many people don’t understand the scope of uses for

bamboo. The shoots can be picked early for eating, and the wood of

older canes can be treated and used as anything from decoration to

instruments. Thankfully, many manufacturers have seen all the

products that can be made from this highly renewable resource and

have begun to utilize bamboo in some fascinating ways.

Decorations: From picture frames to room dividing screens,

bamboo can make some elegant and exotic decorations for the home.

Depending on the manufacturer, bamboo decorations can be the rough

finish of natural bamboo that reminds people of tropical getaways,

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or the sleek, lacquered finish that creates a modern elegance that

many people remember. Bamboo can also be colored so that it can

fit into any décor.

Building materials: More and more furniture, flooring, and

even homes are being built with bamboo. Whether people like the

look of the bamboo, or the way it holds up, it is becoming a more

popular building material that many people are recognizing. The

smooth floors hold up well in kitchens and other rooms, and the

furniture, bound attractively with rattan or leather, gives any

room a modern look.

Fabrics and clothing: A fabulous trend right now is bamboo

fibers being used in fabrics and clothing. Bedding made of bamboo

fibers is as soft as or softer than most cotton beddings, and

drapes with the look of silk without the expense. It is becoming

a mainstream trend to have bamboo fabric products or clothing,

populating many major chain retail stores.

Cooking: Cooking with bamboo is nothing new in Asian culture.

Bamboo shoots are a common food in that part of the world, and

have also migrated into cooking utensils. Bamboo cutting boards

are notoriously good for not dulling blades on knives as quickly,

while bamboo utensils like wooden spoons are excellent for not

scratching the bottoms of expensive non-stick cookware.

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 Agriculture: Bamboo started out as a natural plant in most

places, but has become a large part of agriculture. From being the

main crop of a farm to be harvested for other uses, or as the

channel linings for irrigation systems, bamboo fits naturally into

agriculture. Of course, bamboo is also grown as a food source, and

as a garden plant as well, the woody grass being an excellent

addition to any garden.

Weapons: While this is rarely seen any more, bamboo was once

used to make many different types of weapons. From blow guns to

archery bows and arrows, bamboo made light but strong weapons for

many centuries. Though they aren’t used as frequently any more,

even gunpowder guns have been made with the hollow tubes.

Instruments: Hollow tubes make excellent instruments, whether

it is a flute or a drum, and bamboo is one of the best bases for

instruments. The light, durable quality of the bamboo is coupled

with its musical potential, and creates some of the most beautiful

sounds that music has ever heard.

B. Related Studies

Foreign Literature

Current research reveals the different potential application

of antioxidant/free radical manipulations in prevention or control

of diseases. Natural products from dietary components such as

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Indian species and medicinal plants are known to possess

antioxidant activity. Increasing intake of dietary antioxidants

may help to maintain an adequate antioxidant status and, therefore,

the normal physiological function of a living system. To protect

the cells and organ systems of the body against reactive oxygen

species, humans have evolved a highly sophisticated and complex

antioxidant protection system.

A study conducted by Choi et. al (2018) entitled “Comparison

of Quality Characteristic and Antioxidant Potential of Cultivated

Pu-erh and Gushu Pu-erh Tea Extracts at Two Temperatures”, compared

the pH, titratable acidity, color value, antioxidant potential and

free amino acid contents of cultivated Pu-erh and Gushu Pu-erh tea

extracted at 90 and 100°C. The results of their present study

showed that extraction temperature significantly affect the

chemical and functional values of tea. Gushu Pu-erh showed lower

pH but higher antioxidant potentials and free amino acid content

at both extraction temperatures. The antioxidant potentials and

free amino acid content were high in the tea extracted at 100°C

compared to at 90°C. Based on the nutritional and functional

parameters investigated in the present study, Gushu Pu-erh tea

extracted at 100°C for 3 min with 30 sec of shaking provides high

amounts of total polyphenol, flavonoid and free amino acid.

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 The study by R. Muchacka et al. (2018) entitled “Activity of

Antioxidant Enzymes and Levels of GSH (Gulatathione) and MDA

(Methylenedianiline)in Heritage Breed and Commercial Crosses Hen

Eggs”, determined the activity of antioxidant enzymes and the level

of GSH (Glutathione) and MDA (Methylenedianiline) in yolk and

albumen of eggs laid by 48 and 50-week-old hens. In their

experiment 180 hens of three heritage breeds (Yellowleg Partridge,

Sussex, Leghorn) and 180 commercial crosses of laying hens (Hy-

Line, ISA Brown, Lohmann) were used. Layers were reared on litter

and had no outdoor access. Birds were assigned to treatment groups

I to VI (Yellowleg Partridge, Sussex, Leghorn, and commercial

layers Hy-Line, ISA Brown and Lohmann, respectively). Birds were

fed standard diets based on concentrates for laying hens and

libitum and had free access to water throughout the experiment.

All the groups were managed under uniform environmental (air

humidity and temperature, lighting programme) and feeding

conditions. At 48 and 50 weeks of age, 6 eggs from each group were

collected. In the samples of egg yolks and albumens the

concentration of GSH (Gulatathione) and MDA (Methylenedianiline),

and activity of Superoxide Dismutase (SOD), GPx (glutathione

peroxidase) and CAT (catalase) were estimated. The egg yolks and

egg albumens showed statistically significant differences in SOD

activity, GSH (Gulatathione) and MDA (Methylenedianiline) levels

in both studied periods. At 48 and 50 weeks of age, the highest

15
activity of SOD (superoxide dismutase) and the lowest level of GSH

(Gulatathione) and MDA (Methylenedianiline) were observed in eggs

from Sussex hens (heritage breeds) and in eggs from Hy-Line

(commercial crosses).

In conclusion, it can be stated that the genetic background

of laying hens influences the activity of antioxidant enzymes and

levels of GSH (Gulatathione) and MDA (Methylenedianiline) in eggs,

which may influence their quality. The differences between the

treatment groups in the measurements were maintained in both

studied periods.

The dragon fruit is one of the fruits cultivated in the

tropics. The fruit flesh of the dragon fruit has been widely

consumed, and the fruit peel of the dragon fruit has also been

extensively utilized. But the leaves of the dragon fruit have not

been utilized and tend to be waste in agriculture. The study by

Manurung et. al (2018) entitled “Spectrophotometric Method for

Antioxidant Activity Test and Total Phenolic Determination of Red

Dragon Fruit Leaves and White Dragon Fruit Leaves”, aimed to

utilize waste dragon fruit leaves with the test of antioxidant

activity and the determination of total phenolic of red dragon

fruit leaves extract and white dragon fruit leaves extract by

spectrophotometric method. Methods performed for antioxidant

activity test was by 1,1- diphenyl-2-picrylhydrazyl (DPPH) with

16
ascorbic acid as the comparator and total phenolic determination

by Folin Ciocalteu (FC) with gallic acid as the comparator.

Measurements were done with a spectrophotometer. Antioxidant

activity test results of red dragon fruit leaves extract and white

dragon fruit leaves extract obtained scavenging concentration 50%

(SC50) 135.00 µg/mL and 142.47 µg/mL. Total phenolic determination

results of red dragon fruit leaves extract and white dragon fruit

leaves extract obtained value 756.75 mg/g and 707.07 mg/g. Both

red dragon fruit leaves extract and white dragon fruit leaves

extract to have moderate antioxidant activity. The antioxidant

activity of red dragon fruit leaves is higher than the antioxidant

activity of white dragon fruit leaves. This is because the total

phenolic in red dragon fruit leaves is higher than total phenolic

in white dragon fruit leaves.

In a study by Longe et. al (2014) entitled “In Vitro Studies

on the Antioxidant Property and Inhibition of a-Amylase, a-

Glucosidase and Angiotensin I-Converting Enzyme by Polyphenol-Rich

Extracts from Cocoa (Theobroma cacao) Bean”, the studies of cocoa

and their related products have become an area of interest owing

to their health-promoting properties. In recent years, cocoa and

cocoa products, namely, cocoa powder, dark chocolate, and cocoa

liquor, have been shown to suppress atherosclerosis and reduce the

risk of heart disease increase dermal blood circulation, and

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decrease platelet activation, adhesion, and function, as well as

function as cancer protective agent by inhibiting the

proliferation of human cancer cells and also exerted hypoglycemic

properties owing to the presence of the phenolic compounds in it.

Polyphenols have been researched for decades, mostly because of

their antioxidant properties.

Furthermore, the total phenol and flavonoid contents of the

water extractable phytochemicals from the powdered cocoa bean were

determined and the effects of the extract on α-amylase, α-

glucosidase, and angiotensin-1 converting enzyme activities were

investigated in vitro. In addition, the radicals [1,1-diphenyl-2

picrylhydrazyl (DPPH), 2,2’-azino-bis(3-ethylbenzthiazoline-6-

sulphonic acid) or ABTS, hydroxyl (OH), and nitric oxide (NO)]

scavenging ability and ferric reducing antioxidant property of the

extract were assessed. The results revealed that the extract

inhibited α-amylase (1.81 ± 0.22 mg/mL), α-glucosidase (1.84 ±

0.17 mg/mL), and angiotensin-1 converting enzyme (0.674 ± 0.06 mg/mL

[lungs], 1.006 ± 0.08 mg/mL [heart]) activities in a dose-dependent

manner and also showed dose-dependent radicals [DPPH (16.94 ±

1.34 mg/mL), NO (6.98 ± 0.886 mg/mL), OH (3.72 ± 0.26 mg/mL), and

ABTS (15.7 ± 1.06 mmol/TEAC·g] scavenging ability. Conclusion. The

inhibition of α-amylase, α-glucosidase, and angiotensin-1

converting enzyme activities by the cocoa bean extract could be

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part of the possible mechanism by which the extract could manage

and/or prevent type-2 diabetes and hypertension.

A study conducted by Park et. al (2014) entitled “Antioxidant

Effects of Spinach (Spinacia oleracea L.) Supplementation in

Hyperlipidemic Rats”, aimed to evaluate the antioxidant effects of

spinach in vitro and in vivo in hyperlipidemic rats. For

measurement of in vitro antioxidant activity, spinach was

subjected to hot water extraction (WE) or ethanol extraction (EE)

and examined for total polyphenol content (TPC), oxygen radical

absorbance capacity (ORAC), cellular antioxidant activity (CAA),

and antigenotoxic activity. The in vivo antioxidant activity of

spinach was assessed using blood and liver lipid profiles and

antioxidant status in rats fed a high fat-cholesterol diet (HFCD)

for 6 weeks. The TPC of WE and EE were shown as 1.5±0.0 and 0.5±0.0

mg GAE/g, respectively. Increasing the concentration of the

extracts resulted in increased ORAC value, CAA, and antigenotoxic

activity for all extracts tested. HFCD-fed rats displayed

hyperlipidemia and increased oxidative stress, as indicated by a

significant rise in blood and liver lipid profiles, an increase in

plasma conjugated diene concentration, an increase in liver

thiobarbituric acid reactive substances (TBARS) level, and a

significant decrease in manganese superoxide dismutase (Mn-SOD)

activity compared with rats fed normal diet. However,

19
administration of 5% spinach showed a beneficial effect in HFCD

rats, as indicated by decreased liver TBARS level and DNA damage

in leukocyte and increased plasma conjugated dienes and Mn-SOD

activity. Thus, the antioxidant activity of spinach may be an

effective way to ameliorate high fat and cholesterol diet-induced

oxidative stress.

An investigation by Jegede et. al (2018) in their study

entitled “Evaluation of Antioxidant and Cytotoxic Properties of

Vernonia amygdalina”, was carried out to evaluate the antioxidant

activity and cytotoxic properties of Vernonia amygdalina. The free

radical scavenging activity using a stable radical; 2, 2-Diphenyl-

1-picryl hydrazyl, lipid peroxidation assay (DPPH), and nitric

oxide inhibitory assay gave the highest percentage inhibition as

74.55 ± 1.07 %; IC50 = 1.831, 60.42±0.11; IC50 = 3.84 ± 1.03 and

71.26±0.48; IC50 = 0.99 mg/ml, respectively. This is comparable to

the standards quercetin used (P > 0.05). In addition; total phenol,

total flavoniods, anthocyanin and proanthocyanidine of the extract

were determined using established methods. The results obtained

justify the scavenging activity of the extracts. Furthermore, the

extracts possessed very low cytotoxicity to brine-shrimp lethality

test, when compared with the reference standard (Potassium

dichromate, LC50 = 0.003 ± μg/ml). The results obtained in theie

study indicate that V. amygdalina can be a safe potential source

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of natural antioxidant agent; used as a neutralcetical/functional

food.

Yi-ZhongCai et. al (2013) stated in their study entitled

“Antioxidant Activity and Nutritional Quality of Traditional Red-

Grained Rice Varieties Containing Proanthocyanidins”, that

proanthocyanidin-containing rice varieties have been rarely

reported. Antioxidant capacity, major antioxidant components, and

nutritional parameters of eight traditional red-grained rice

varieties containing proanthocyanidins grown in Sri Lanka were

investigated. The tested traditional red varieties, on the

average, had over sevenfold higher both total antioxidant capacity

and phenolic content than three light brown-grained new-improved

rice varieties. Major antioxidant phenolic compounds identified in

this study included proanthocyanidins, phenolic acids and γ-

oryzanols (ferulic acid derivatives). Proanthocyanidins were

detected only in the traditional red varieties, but not found in

new-improved ones. Most traditional red varieties also contained

significantly higher levels of protein with well balanced amino

acids and higher contents of fat, fibre and vitamin E (tocopherols

and tocotrienols) than the new-improved ones. Great variations in

antioxidant capacity, major phenolics, and nutritional parameters

were observed among different rice varieties.

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 A study by Omede (2016) entitled “Total Polyphenolic Content

and Antioxidant Properties of Moringga oliefera Leaf Extracts”,

was carried out to evaluate the relative antioxidant properties

and polyphenol contents of partially purified fractions of Moringa

oleifera leaves extracts. The total phenolic, total flavonoid,

anthocyanin, proanthocyanidine and tannin contents of the crude

methanolic extract, aqueous fraction and ethyl acetate fraction

were determined using established methods, while the antioxidant

properties of the test fractions were evaluated using five in vitro

radical scavenging assays: 2,2-diphenyl- 1-picrylhydrazyl (DPPH)

radical scavenging assay, nitric oxide inhibitory assay, lipid

peroxidation assay, reductive potential assay, and the ferric

reducing ability of plasma (FRAP) assay. The highest radical

scavenging effect and polyphenol contents were observed in the

ethyl acetate fractions than the other fractions: the order of

activity for all the assays was ethyl acetate reaction > crude

extract > aqueous extract. The results obtained in the present

study indicates that M. oleifera could be a potential source of

natural antioxidant and could be applied as a functional food as

regard its relatively low tannin content.

In a study by Hwang et. al (2014) entitled “Phenol Content,

Antioxidant and Tyrosine Inhibitory Activity of Mangrove Plants in

Micronesia”, mangrove samples were harvested at the shoreline on

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the island of Weno, Chuuk State in Micronesia. The phenol content,

antioxidant activity (based on DPPH-free radical scavenging) and

tyrosinase inhibitory activity in different tissues (leaves, barks

and roots) of Rhizophora stylosa (R. stylosa) and Sonneratia alba

(S. alba), collected from the island of Weno. They compared the in

vitro antioxidant and tyrosinase inhibitory activities of two

species of mangrove plants. Total phenol content ranged from 4.87

to 11.96 mg per g of freeze dried samples. The highest antioxidant

activity was observed in R. stylosa bark (85.5%). The highest

tyrosinase inhibitory activity was found in S. alba bark. Also,

total phenol content and antioxidant activity were higher in

methanol extracts than in aqueous extracts. Taken together, the

results of their study proved that mangroves can be excellent

sources of antioxidant compounds.

In a study conducted by Kingsley et. al (2016) entitled

“Antioxidant and Antidiabetic Activities of the Seed and Leaf

Extracts of Chrysophyllum albidum”, they investigated the

antioxidant and antidiabetic activities of the seed and leaf

extracts of Chrysophyllum albidum (C. albidum). After assessing

the in vitro ferric reducing power and hydrogen peroxide scavenging

activities as well as the flavonoid and flavanol contents, the

seed and leaf extracts were administered to diabetic rats for 7

days. The animals were sacrificed and serum was obtained for the

23
determination of blood glucose level while liver sample was used

for the quantification of glycogen level as well as lipidic

peroxidation and catalase activity. Seed and leaf extracts of C.

albidum showed ferric reducing activity and very high hydrogen

peroxide scavenging potential. After the administration of

treatment in diabetic rats, there was a significant decrease (P

<0.05) in blood sugar level and a significant increase (P<0.05) in

liver glycogen level in Groups 3 and 4 animals administered the

leaf and seed extracts respectively compared to Group 1 (the

negative control). Also, catalase activity and malondialdehyde

levels increased in Groups 3 and 4 administered the extracts

compared to Group 1 animals (the negative control). Flavonoids and

flavanol were present and significantly higher (P<0.05) in the

leaf than seed extract. In all, the leaf extract showed the

greatest activities. These results suggest that the leaf and seed

extracts of C. albidum possess both in vitro and in vivo

antioxidant activities in scavenging free radicals as well as

antidiabetic activity, and as such, a potentially important

compound in antidiabetic drug discovery.

Local Literature

According to a study by Geronimo et. al (2013) entitled

“Philippine Yam (Dioscorea spp.) Tubers Phenolic Content and

Antioxidant Capacity”, five Philippine varieties of purple yam

24
or ube (Dioscorea alata) — Daking, Kimabajo, Rapang-rapang,

Sampero, and Shiket, and two varieties of lesser yam or tugui

(Dioscorea esculenta)— Highland and Lowland, were analyzed in

their study for phenolic content and antioxidant activity. The

total phenolic content of the samples ranged from 69.9 to 421.8 mg

gallic acid equivalent (GAE)/100 g dry weight. EC50 values were

1.7-14.8, 6.2-31.7, and 17.5-35.1 mg/mL for radical scavenging

activity, reducing power and iron chelating capacity,

respectively. Total antioxidant activity by ferric thiocyanate

method at 50 mg/mL was between 92.0-95.6%. All samples had better

radical scavenging activity and reducing power on a µg analyte

basis than α-tocopherol. Significant correlation was observed

between total phenolic content and DPPH radical scavenging

activity (R=-0.7664, p<0.05) and reducing power (R=-

0.8083, p<0.05) but none between total antioxidant activity and

phenolic content (0.1378, p>0.05), for both purple yam and tugui.

Significant correlation between total phenolic content and iron-

chelating capacity was observed only for the tugui varieties (R=-

0.9859, p<0.05).

According to a study by Cariscal et. al (2017) entitled

“Selected Philippine Plant Extracts as Alternative Preservatives

for a Pharmaceutical Liquid Preparation”, synthetic or chemical

preservatives, such as bisulfites, nitrates, butylated

25
hydroxytoluene (BHT), benzoic acid, methylparaben and

propylparaben, have been used for decades in pharmaceutical

preparations (Abdel-Azi et al. 2016) and thus, are readily

available. Their mechanisms of action are based on their

antioxidant, antimicrobial or anti-enzymatic properties. However,

various studies have been conducted which demonstrated their

ability to cause undesirable effects such as allergic reactions

(Wilson and Bahna 2005), carcinogenicity (Gharavi et al. 2007),

behavioral changes (McCann et al. 2010) and even mental

performance.

Preservatives play an essential role in enhancing quality and

prolonging shelf-life of pharmaceutical products by improving

their antimicrobial stability or reducing the amounts of oxidative

degradation products. Persistent use of synthetic compounds as

preservatives resulted in several reports of undesirable effects.

Hence, development of alternatives is necessary to maintain their

vital function while minimizing adverse effects. In their study,

ethanolic extracts of five plants with known antimicrobial

activities, Psidium guajava, Premna odorata, Mimosa pudica, Allium

sativum and Zingiber officinale, were formulated into suspensions

and evaluated for preservative activity using the United States

Pharmacopeia (USP) (2015) guidelines. Phytochemical test,

antioxidant activity and compatibility test were also conducted on

26
the extracts. Premna odorata (p=0.999) and Mimosa pudica (p=0.054)

at 5.00 mg/mL concentration exhibited comparable antioxidant

activity against the standard antioxidant preservative, butylated

hydroxytoluene, using ferric reduction antioxidant power assay.

Based on the criteria for product category 4 of the USP,

suspensions of Premna odorata and Psidium guajava demonstrated

acceptable preservative activity against selected microorganisms,

Escherichia coli and Staphylococcus aureus. These bioactivities

can be attributed to the phytochemicals present in the extracts

such as glycosides, reducing substances, flavonoids and alkaloids.

In conclusion, for the USP category 4 products such as antacid

suspensions, Psidium guajava can be utilized as an alternative

source of antimicrobial preservative, Mimosa pudica as an

alternative source of antioxidant preservative, and Premna odorata

as an alternative source of preservative with both antimicrobial

and antioxidant efficacy.

A study by Uy et. al (2016) entitled “Screening of the

Antioxidant Properties of the Leaf Extracts of Philippine

Medicinal Plants Ficus nota ( Blanco ) Merr. Metroxylon sagu

Rottb., Mussaenda philippica A. Rich., Inocarpus fagifer, and

Cinnamomum mercadoi Vidal”, continued search for natural

antioxidants from the decoction and ethanol extracts of the leaves

of Ficus nota (Blanco) Merr., Metroxylon sagu Rottb., Mussaenda

27
philippica A. Rich, Inocarpus fagifer, and Cinnamomum mercadoi

Vidal. The C. mercadoi ethanolic (CmE) extract demonstrated the

highest amount of total phenolics (570.58 mg GAE) and it correlates

well to its strong radical scavenging activity (91.97% at 500 ppm)

against 1,1Diphenyl-2-picrylhydrazyl (DPPH) and high antioxidant

capacities (149.91 AAE, 209.98 BHTE). The M. sagu decoction (MsD),

(403.00 mg GAE, 96.39 AAE, 139.61 BHTE) and ethanolic (MsE),

(310.58 mg GAE/g, 73.49 AAE, 102.36 BHTE) extracts are the second

and third that showed high amount of phenolics, strong free radical

scavenging against DPPH and high antioxidant capacities,

respectively. These are followed by decoction extracts of F. nota

(FnD) (88.19% at 500 ppm against DPPH, 216.64 mg GAE, 55.52 AAE,

77.05 BHTE), I. fagifer (IfD) (83.64% at 500 ppm against DPPH,

194.48 mg GAE, 63.13 AAE, 88.89 BHTE), C. mercadoi (CmD) (83.64%

at 500 ppm against DPPH, 166.03 mg GAE, 80.88 AAE, 88.31 BHTE),

and ethanolic extract of I. fagifer (IfE) (89.76% at 500 ppm

against DPPH, 131.48 mg GAE, 59.48 AAE, 82.63 BHTE). Their findings

may support their traditional/ethno- medicinal claims. Their study

further indicates that the extracts from C. mercadoi, M. sagu, F.

nota, and I. fagifer can be used as important sources of natural

antioxidants that may offer protection from the harmful effects

caused by overproduction of radicals in the body.

28
 In a study conducted by Palmes et. al (2015) entitled

“Phytochemical Profiles and Antioxidant Activity of Selected

Indigenous Vegetables in Northern Mindanao, Philippines”, the

crude methanol extracts of five indigenous vegetables namely,

Amarathus tricolor, Basella rubra L., Chochurus olitorius L.,

Ipomea batatas, and Momordica chuchinensis L., were examined for

their phytochemical profile and antioxidant activity using 1,1-

diphenyl-2-picrylhydrazyl (DPPH) free radical. The values for DPPH

radical scavenging activity ranged from 7.6-89.53% with B. rubra

and I. batatas having the lowest and highest values, respectively.

The total flavonoid content of all five indigenous vegetables

ranged from 74.65-277.3 mg quercetin equivalent per gram of dried

vegetable material while the total phenolic content ranged from

1.93-6.15 mg gallic acid equivalent per gram dried material.

Phytochemical screening revealed the presence of steroids,

flavonoids, saponins, tannins, carbohydrates and reducing sugars,

which may also be associated with the antioxidant activity shown

by these indigenous vegetables.

According to a study conducted by Rivera et. al (2015)

entitled “Free-Radical Scavenging Activity and Bioactive Secondary

Metabolites from Various Extracts of Glinus oppositifolius (L.)

Aug. DC. (Molluginaceae) Roots, Stems and Leaves”, the plant

belonging to the family Molluginaceae, is a slender or ascending,

29
smooth, branched, annual herb, with branches 10-40 centimeters

long. A well-renowned academic communication platform for the rest

of the world on tropical medicine and other related fields,

reported that G. oppositifolius has therapeutic benefits in

traditional medicine. Among them include its analgesic,

antidiabetic, anti-hyperlipidemic, antihelminthic,

antidiarrhoeal, diuretic, antimalarial, antiviral, antimicrobial

and antioxidant properties. The shoot of G. oppositifolius is eaten

occasionally as a vegetable even though it is bitter on account of

its stomachic, aperient, and antiseptic properties. The whole

plant, without the roots, is used as a cooked cataplasm in

dyspepsia in children and as an infusion to promote the menstrual

discharge in women. It is used as a blood purifier and liver

stimulant. It can also improve digestion and can cure burning

sensation, itchiness and other skin ailments. The experts from UP

reported that the antioxidant activity of various solvent

(ethanol, methanol, chloroform) extracts come from different parts

of G. oppositifolius, including its roots, stems, and leaves.

In a study conducted by Bungihan et. al (2013) entitled

“Determination of the Antioxidant Phytochemical and Antibacterial

Profiles of Flowers from Selected Ornamental Plants in Neuva

Vizcaya, Philippines”, stated that the determination of the

Antioxidant, Phytochemical and Antibacterial Profiles of Flowers

30
from Selected Ornamental Plants in Nueva Vizcaya, Philippines

Different flowers can be utilized for their antibacterial and

antioxidant values. They have the potential for pharmaceutical and

nutraceutical development and in cosmetics. However, there is a

need to assess their toxicological and pharmacological effects and

establish their safety. The results of their study are important

as preliminary for future studies. Furthermore, there are still a

lot of flowers which could be given consideration. This also

established scientific basis of the folkloric claims about the

uses of the flowers of some of the plants.

With the increasing interest on natural products discovery,

utilization of plant parts which are commonly disregarded is

another breakthrough. Their study sought to determine the

secondary metabolites present, phenolic contents, radical

scavenging activities and antibacterial properties of 10 selected

ornamental flowers which were Adenium obesum, Allamanda

cathartica, Bougainvillea glabra, Catharanthus roseus, Caesalpinia

pulcherrima, Heliconia subulata, Ixora coccinea, Mussaenda

philippica, Tecoma stans and Torenia fournieri, from Nueva

Vizcaya, Philippines. The ethanoliccrude extracts were subjected

to thin layer chromatography (TLC) phytochemical screening. Total

phenolics expressed as mg AAE/g (Ascorbic Acid per gram) sample

were determined through Folin-Ciocalteu method. Radical scavenging

31
activities were measured using DPPH assay. Antibacterial

activities were assessed through TLC bioautography. To quantify

bacterial susceptibility of C. pulcherrima, Minimum Inhibitory

(MIC) and Minimum Bacterial Concentration (MBC) were employed in

the ethyl acetate sub-fraction. TLC showed different types of

phenolics, alkaloids, essential oils and terpenes. T. fournieri

showed the highest total phenolics with 155 mg AAE/g (Ascorbic

Acid per gram) while H. subulatahad showed the lowest with 3.50 mg

AAE/g (Ascorbic Acid per gram). Measurement of IC50 showed that H.

subulata had the highest antioxidant activity followed by T.

fournieri with 303 ppm and 320 ppm, respectively. H. subulata

having the least amount of total phenolics had greatest radical

scavenging activity. This indicates that other than phenolics,

other metabolites can pose great antioxidant activities. The

antibacterial testing showed that M. philippica, I. coccinea, H.

subulata, C. pulcherrima and A. catharticaare were bioactive

against Staphylococcus aureus and Escherichia coli. MIC and MBC of

C. roseus.

According to a study by Barcelo (2015) entitled

“Phytochemical Screening and Antoxidant Activity of Edible Wild

Fruits in Benguet, Cordillera Administrative Region, Philippines”,

it identified the secondary metabolites present and determined the

antioxidant activity of 31 edible wild fruits grown in Benguet

32
province, Cordillera Administrative Region, Philippines. Total

polyphenol and flavonoid content were estimated using Folin-

Ciocalteu and aluminium chloride method respectively. Antioxidant

activity was measured through diphenyl-1-picrylhydrazyl assay.

Based on the results of the study the following bioactive

constituents are present in the fruits: alkaloids, steroid

glycosides, saponins, flavonoids, polyphenols and tannins. The

fruits contain more polyphenols than flavonoids. All the fruits

except Physalis peruviana (Solanaceae) exhibited higher

antioxidant activity than Vitamin E (Myra E), ascorbic acid (50

ug/mL), and trolox (1000 uM). Dillenia philippinensis

(Dilleniaceae) exhibited the highest antioxidant activity. The

antioxidant activity of the fruits and controls is significantly

different (ρ ≤ 0.05). Post-hoc Tukey analysis of data reveals that

several fruits have equal activity. Finally, there is a positive

moderate correlation (r=0.50) between the total polyphenol content

and antioxidant activity of the fruits.

Statement of the Problem

This study aimed to determine the antioxidant activity of

Golden Buho (Schizostachyum brachycladum) leaves extract in 2,2-

diphenyl-1-picrylhydrazyl(DPPH).

33
 Specifically, it sought to answer the following questions:

1. What are the secondary metabolites present in the leaves of

Golden Buho (Schizostachyum brachycladum)?

2. Is there an antioxidant activity in the test drug that reacts

in the DPPH in a concentration level of 200 ppm (parts per

million) to 1000 ppm (parts per million)?

3. Is there a significance difference in the antioxidant

activity of the test drug and the positive control?

Hypothesis

Null Hypothesis:

1. There is no antioxidant activity in the test drug that reacts

in the DPPH in a concentration level of 200 ppm (parts per

million) to 1000 ppm (parts per million).

2. There is no significant difference in the antioxidant

activity of the test drug and the positive control.

Conceptual Framework

INDEPENDENT VARIABLE DEPENDENT VARIABLE

• Schizostachyum • Reduction of
Brachycladum DPPH
bamboo leaves
extract
Figure 1: Research Paradigm

34
 The study’s conceptual framework shows the relationship of

variables in this study. The independent variable in the study is

the extract from Golden Buho (Schizostachyum brachycladum) leaves

and as our experimental group to determine our dependent variable

which is the reduction of DPPH (2,2-diphenyl-1-picrylhydrazyl).

Significance of the Study

The research work aimed to determine the effectiveness of the

Golden Buho (Schizostachyum brachycladum) leaves extract in

reducing the free radical level of the body.

The findings of the research work are beneficial for the

following:

Patients. This study will be beneficial for the patients

because it provides natural and cheap medicine that can help

minimize the multiplication of cancer cells, and age dependent

diseases. This also help patients in minimizing their expenses

used on buying commercial drugs.

Health Sectors. This study will be beneficial for the health

sectors for they can study and widen their ideas and gain new

knowledge about antioxidants activity of Golden Buho

(Schizostachyum brachycladum) leaves extract as a medicine which

is effective and inexpensive.

35
 Community. This study will be benefical in a community, in

such a way that it gives solution to the problem of the people in

aging, and age dependent diseases such as cardiovascular disease,

cancer, neurodegenerative disorders, and other chronic conditions.

The Researchers. It will be also beneficial to the researchers

not only in gaining intellectual advancements about the

effectiveness and applicability of the Golden Buho (Schizostachyun

brachycladum) leaves extract to their health. This study will also

help the researchers about the other health benefits that they may

do in a related situation that many reserchers were not be able to

explore or know.

Future Researchers. This study will be beneficial for the

future researchers who are interested to conduct similar study.

This study will serve as their reference that will give them ideas

for further information to create new research study. They can

also use this study as an additional literature for their future

research works.

Scope and Delimitation

This study focused on antioxidant activity of Golden Buho

(Schizostachyum brachycladum) leaves extract in 2,2-diphenyl-1-

picrylhydrazyl(DPPH). The DPPH was bought from the Sigma Aldrich

(Tanza, Quezon City). The Golden Buho leaves was harvested in

36
Solana, Cagayan. In connection to this, plant authentication was

done at Department of Environment and Natural Resources Region II.

Definition of Terms

This study entitled on “Antioxidant Activity of Golden Buho

(Schizostachyum brachycladum) Leaves Extract in 2,2-diphenyl-1-

picrylhydrazyl(DPPH)” used the following terminologies:

Free radical – an uncharged molecule (typically highly reactive

and short-lived) having an unpaired valence electron which

scavenge the body to seek out other electrons so they can become

a pair which causes damage to cells, proteins and Deoxyribonucleic

acid (DNA).

Antioxidant – a substance that inhibits oxidation which prevents

the chain reaction of free radicals in the body.

DPPH - a kind of free radical that will be measured in this study.

Schizostachyun brachycladum – type of a plant that is being analyze

to determine its antioxidant activity.

UV-VIS Spectrophotometer - device used to determine the

antioxidant activity of the test samples.

37
 CHAPTER II

METHODOLOGY

This chapter deals with the methods and procedure to determine

the antioxidant activity of golden buho (Schizostachyum

brachycladum) leaves extract in 2,2-diphenyl-1-

picrylhydrazyl(DPPH).

Research Design

The design that was used in this study was the experimental

research design.

A. Chemicals

DPPP were purchased from Sigma Aldrich (Tanza, Quezon City).

It served as the reagent of this study.

Ascorbic acid, which was provided by the Central Analytical

Laboratory of Cagayan State University as part of the laboratory

service, was used to reduce the DPPH and served as the positive

control of this study.

Ethanol was also used to dilute the DPPH, Ascorbic acid, and

the Golden Buho. It was also provided by the Central Analytical

Laboratory of Cagayan State University as part of the laboratory

service. Ethanol also served as the negative control of this study.

38
B. Plant Materials

The leaves of Golden Buho (Schizostachyum brachycladum) were

gathered from Centro, Solana, Cagayan. A plant sample was brought

to the National Plant Quarantine Services Division for

authentication.

C. Phytochemical Analysis

Golden Buho leaves extract was brought to the Department of

Science and Technology, Regional Standard and Testing Laboratory

in Bagay, Tuguegarao City for Phytochemical screening to determine

the secondary metabolites present in the extract.

D. Preparation of DPPH, Ascorbic acid, and test drug samples

1. DPPH was stored in a cool dry place to maintain its

temperature.

2. A stock solution of 1.0 mM (millimolar) was prepared. 0.1

gram of DPPH was weighed then it was diluted to 250 ml with

ethanol, making 0.1 mM (millimolar) of DPPH.

3. From stock, 25ml of 1.0 mM (millimolar) DPPH solution was

measured and diluted to 250 ml of ethanol.

4. 200 mg of sample was weighed and diluted to 250 ml ethanol

producing 1000 ppm (parts per million) of the test drug

sample.

39
5. Same preparations of Golden Buho leaves extract and Ascorbic

Acid were as follows:

a. 800 ppm (parts per million): an aliquot of 40 ml from

the 1000 ppm (parts per million) of the test sample was

diluted to 50ml ethanol.

b. 600 ppm (parts per million): an aliquot of 30 ml from

the 1000 ppm (parts per million) of the test sample was

diluted to 50ml ethanol.

c. 400 ppm (parts per million): an aliquot of 20 ml from

the 1000 ppm (parts per million) of the test sample was

diluted to 50ml ethanol.

d. 200 ppm (parts per million): an aliquot of 10 ml from

the 1000 ppm (parts per million) of the test sample was

diluted to 50ml ethanol.

6. A 1 ml of 0.1 mM DPPH solution plus 3 ml of each test sample

and ascorbic acid was added at different concentration. It

was protected from light by covering the test tubes with

aluminum foil and stored in dark area for 30 mins. The reading

of absorbance was done to a device called UV-Vis

Spectrophotometer at a wavelength of 517 nm (nanometer).

40
E. Plant Extraction

Golden Buho leaves were gathered, washed, and air dried

thoroughly. The ethanolic extraction was done by soaking 200 grams

of Golden Buho leaves in 1930 ml of ethyl alcohol for 24 hours.

After soaking, the leaves of Golden Buho were filtered using a

filter paper. The collected ethanol extract undergone Rotary

Evaporation. And the ethanol extract was placed in a clean

container.

F. Statistical Tool

The data were subjected to Paired T-test with a 5% level of

significance to compare the antioxidant activity of the test drug

and positive control.

41
 CHAPTER III

RESULTS AND DISCUSSION

This chapter presents the results, data gathered, in

accordance to the antioxidant activity of Golden Buho

(Schizostachyum brachycladum) leaves extract in 2,2-diphenyl-1-

picrylhydrazyl(DPPH). The data is discussed through tables and

line graphs.

Table 1. Result of the Phytochemical Analysis of Golden Buho

(Schizostachyum brachycladum) Leaves Extract

Sample
Parameter Result Method Used
Description

Phytochemical
Screening:

Alkaloids -

Anthocyanins -

Flavonoids +

Phenolics +
Golden Buho Guevarra et al
Leaves extract (2005)
Reducing
-
Sugars

Saponins +

Steroids -

Tannins +

Terpenoids +

42
 As seen in the table above, the phytochemical analysis of

Golden Buho leaves extract showed the presence of Flavonoids,

Phenolics, Saponins, Tannins and Terpenoids.

Figure 1. Comparison of Golden Buho (Schizostachyum brachycladum)

and Ascorbic Acid in their percent inhibition in different

concentration

Percent Inhibition in Different Concentration

50
6.25 9.375 12.5 14.0625
3.125
0
200 400 600 800 1000
-50 -32.8125
-42.1875
%RSA

-100
-96.875
-114.0625
-150

-200
-190.625

-250
Concentration (ppm)

Golden Buho Ascorbic Acid

The figure above shows the comparison of Golden Buho

(Schizostachyum brachycladum) and Ascorbic Acid in their percent

inhibition in different concentration. In getting the %RSA the

𝑨𝒐−𝑨𝒔
formula %𝑹𝑺𝑨 = 𝒙 𝟏𝟎𝟎 (where: Ao = Absorbance of blank/negative
𝑨𝒐

control (0.064); As = Absorbance of sample) was used in this study.

43
 In Golden Buho (Schizostachyum brachycladum), at 200 ppm

(parts per million) the %RSA (Radical Scavenging Activity) is -

190.625%. At 400 ppm, the equivalent %RSA is -114.0625%. At 600

ppm, the equivalent %RSA is -96.875%. At 800ppm, the equivalent

%RSA is -42.1875%. At 1000ppm, the equivalent %RSA is -32.8125%.

Which shows that Golden Buho has negative inhibition in different

concentration ranging from 200 ppm to 1000 ppm.

To sum up, there was no antioxidant activity found in Golden

Buho leaves extract.

Table 2. Comparison of %RSA (Radical Scavenging Activity) by using

the Paired T-test.

Treatment Std. T-Test

Mean P-value Decision
Group Deviation Value
Golden Buho -95.312500 63.5747438
Reject Ho
-3.942 0.017
Ascorbic Acid 9.062500 4.4743191 (Significant)

*Significant at 0.05

Table 2 reveals that the null hypothesis is rejected because

the p-value, 0.017, is less than the level of significance, 0.05,

which means that there is a significant difference in the

antioxidant activity of Golden Buho and Ascorbic Acid.

44
 CHAPTER IV

SUMMARY, CONCLUSIONS AND RECOMMENDATIONS

This chapter presents the summary of the findings, the

conclusions that can be derived from the data sets and the

recommendations that the researchers formulates during the finding

of antioxidant activity of Golden Buho (Schizostachyum

brachycladum) leaves extract in 2,2-diphenyl-1-

picrylhydrazyl(DPPH).

A. Summary

This study aimed to determine the antioxidant activity of Golden

Buho (Schizostachyum brachycladum) leaves extract in 2,2-diphenyl-

1-picrylhydrazyl(DPPH). The DPPH (2,2-diphenyl-1-picrylhydrazyl)

and Golden Buho (Schizostachyum brachycladum) leaves extract were

diluted in an ethanol. The DPPH solution was mixed in Golden Buho

(Schizostachyum brachycladum) leaves extract solution with varying

concentration ranging from 200 ppm to 1000 ppm. The results were

done through a device called “UV-Vis Spectrophotometer”. It was

found out that the Golden Buho (Schizostachyum brachycladum)

leaves extract has no antioxidant activity at a concentration level

of 200 ppm to 1000 ppm.

45
B. Conclusion

Based on the findings presented on the above line graphs, the

researchers now can conclude that the null hypothesis will be

accepted. This means that there is no antioxidant activity in

Golden Buho (Schizostachyum brachycladum) leaves extract that

reacts in the DPPH basing on the %RSA where the results were all

negative starting from 200 ppm up to 1000 ppm.

C. Recommendations

In the light of the findings and conclusion of this research

the following recommendations are given.

1. A similar study should be conducted using other parts of

Golden Buho (Schizostachyum brachycladum) (e.g. bamboo shoots

or the bamboo culms).

2. Consider searching for other protocols of DPPH (2,2-diphenyl-

1-picrylhydrazyl) and Golden Buho (Schizostachyum

brachycladum) which concentration is higher than 1000 ppm for

best result.

3. Try to find an alternative method for determining the DPPH

absorbance.

4. Consider using another type of reagent.

46
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