Extracts of Citrofortunella microcarpa(calamansi) as hand sanitizer against

Hand Bacterias

A Research Proposal Presented to the

Science, Technology, Engineering and, Mathematics (STEM) Department

Central Philippine University Senior High School

Iloilo City

In Partial Fulfillment

Of the requirement for the subject

Research II

Ralfh Lourenze Juelar

Joshua Emmanuel Huervana

Dennis Earl Oriemo

Gabriel Shian Llenares

Jan Karsten Macoco

Researchers
 Chapter 1

Introduction to the Study

Chapter one has five main parts:(1) significance of the study, (2) statement of the

problem and hypothesis, (3) significance of the study,(4) Definition of Terms, and (5) Scope and

limitations of the Study.

Part One, Background of the Study and Conceptual Framework of the Study, presents

an introduction or overview of the present investigation. It explains the rationale of the study,

theories formerly conducted by scholars from whom the research study is based and discussed

the reason why there is a need for the study to be conducted.

Part Two, Statement of the Problem and Hypothesis, describes the main purpose for

conducting the study and list down specific objectives which the study seeks to answer. It also

states the hypothesis that is to be tested in the study.

Part Three, Significance of the Study, states the benefits that could be derived from the

findings of the study.

Part Four, Definition of Terms, list down alphabetically the difficult terms included in the

study and provides the conceptual and operational definition for the purpose of clarity and

understanding.

Part Five, Delimitations of the Study, specifies the coverage of the investigation in terms

of purpose, variables, subjects, research design and methodologies, research instrument used,

and the statistical tools and analytical scheme.

Citrofortunella microcarpa ,Bunge 1832 (Calamansi) is a significant citrofortunella, implying

thatit is an intergenetic half breed between an individual from the variety citrus (for this situation

most likely the mandarin orange) and the kumquat having a place with Fortunella. It has an

impressive measure of unadulterated concentrate put away in the skin. The most widely

recognized techniques for extraction are the cool press strategy, maceration strategy, and

outward extraction. The calamansi unadulterated concentrate is an all-characteristic personal

stench eliminator and antiperspirant. Likewise, calamansi concentrate improves the excellence

and nature of skin and hair.

A hand germ-free, hand disinfectant, hand sanitizer or hand sanitizer is an enhancement or

choice to hand washing with cleanser and water. Numerous arrangements are accessible,

including gel, froth, and fluid arrangements. The dynamic fixing close by sanitizers might be

isopropyl liquor (isopropanol), ethanol, n-propanol, or povidone-iodine. Idle fixings in liquor rub

regularly incorporate a thickening operator, for example, polyacrylic corrosive for liquor gels,

humectants, for example, glycerin for fluid rubs, propylene glycol, and basic oils of plants.

Liquor-based hand sanitizers are more successful at slaughtering microorganisms than

cleansers and don't dry out hands to such an extent.

The reason for making an antibacterial hand sanitizer from calamansi (citrofortunella microcarpa

,Bunge 1832) unadulterated concentrate is to make another substance and to find if this works,

to help clean the general population's hands and to be without germ.

Calamansi fruit/Calamansi

Calamansi extract

Citric Acid

Compound that can be used in

making Santizers
 Figure1.1 Flowchart showing the potential of the fruit extract of the calamansi as a

sanitizer. Calamansi( citrofortunella microcarpa ,Bunge 1832) is a citrus fruit, with a

sour flavour said to resemble a cross between mandarin and lime. As seen on

Figure1.1 its fruit extract has citric acid which is a common ingredient in sanitizers as an

anti oxidant..

Statement of The Problem

The primary motivation behind this examination is to use Calamansi Pure Extract as an

antibacterial hand sanitizer.

In particular, it means to address the accompanying inquiries:

1. Is there a huge distinction between the hand sanitizer in fluid structure with Calamansi Pure

Extract, hand sanitizer in gel structure with Calamansi Pure Extract and popularized hand

sanitizer as far as the accompanying:

1.1 Texture;

1.2 Smell; and

1.3 Appearance?

2. Is there a huge connection between the grouping of Calamansi Pure Extract and the

estimation of zone restraint of Propionibacteria?

3. Is there a huge connection between the grouping of Calamansi Pure Extract and the

estimation of zone restraint of Dermobacteria?

1. There is no noteworthy distinction between the hand sanitizer with Calamansi Pure Extract as

far as the accompanying:

1.1 Texture;

1.2 Smell; and

1.3 Appearance?

2.There is no noteworthy connection between the measure of Calamansi Pure Extract on the

hand sanitizer as far as the accompanying:

2.1 Texture;

2.2 Smell; and

2.3 Appearance?

3. There is no significant relationship between the amount of Pure Extract, propionibacteria and

the dermobacteria bacteria zone of inhibition.

Definition of Terms

Antibacterial properties – is what the researchers will test after making the product.

Calamansi pure extract– to be used for the product.

Calamansi rind – is where the reasearchers will get the calamnsi pure extract.

Hand sanitizer – is what the researchers will make with the calamansi essential oil.

Maceration – is a method pretains to the preperation of an extract by solvent extraction.

This investigation intends to deliver an antibacterial hand sanitizer from calamansi

unadulterated concentrate. The item will contain humectants (glycerin), emollients, emulsifiers,

sterilizers (isopropyl liquor) and calamansi unadulterated concentrate. The examination will take

about a month or something like that, because of different antibacterial tests and testing. The

investigation will happen at the researchers houses and Central Philippine University.

The researchers will get the calamansi unadulterated concentrate carefully from the organic

product's peelings, and not from the natural product's seeds, leaves or tissue. The specialists

will get the calamansi unadulterated concentrate through methods for maceration.
 CHAPTER II

REVIEW OF RELATED LITERATURE

This chapter presents the review of related literature on the , following topics :

Calamansi(Citrofortunella microcarpa, Bunge 1832), hand sanitizers and, hand bacteria a

synthesis of the review is also presented.

The researchers are creating a more effective hand sanitizer with ingredients that are

usually found locally. Calamansi is an example of this local ingredient that is needed for hand

sanitizer. Calamondin (Citrofortunella microcarpa or Citrofortunella mitis) is an important

citrofortunella, meaning that the Calamondin is an intergenetic hybrid between a member of the

genus citrus (in this case probably the mandarin orange) and the kumquat belong to Fortunella.

(Potonia, 2014)

Calamondin (Citrofortunella microcarpa, Bunge 1832) is a shrub or small tree growing to

3–6 meters (9.8–19.7 ft). The plant is characterized by wing-like appendages on the leaf stalks

and white or purplish flowers. The fruit of the calamondin resembles a small, round lime, usually

25-35 mm in diameter, but sometimes up to 45 mm. The center pulp and juice is the orange

color of a tangerine with a very thin orange peel when ripe. Each fruit contains 8 to 12 seeds.

(Potonia, 2014)

Calamansi extract is composed of ascorbic acid, phosphorus and calcium – the extract

is derived from a fruit tree that is native to the Philippines. Today, women in the Philippines (and

around the world) use Calamansi extract to improve the beauty and quality of skin and hair. The

extract may be found in skin lotions, such as body lotions or cold creams, or added to

shampoos and conditioners. Many women in the Philippines find the extract of Calamansi an

important element of staying youthful looking and growing long, shiny hair that is both strong

and glossy.(Kingston. 2011)

such as Papaya extract. The combination of Calamansi and other fruit acids are believed to help

the skin shed dead skin cells and reveal a smoother, rosier and less wrinkled surface. Soaps

made with Calamansi may be designed for use on the body, or especially for the facial area.

Sometimes, these soaps will be used alongside toners and moisturizers that also contain

Calamansi.(Rowland, 2014)

Shampoos, conditioners and hair products containing Calamansi may be used to clean,

condition and groom the hair. These products will use small percentages of Calamansi extract,

alongside larger concentrations of typical shampoo, hair conditioner and hair product

ingredients, such as aloe vera extract and mild detergents.(Rownland, 2014)

Lotions containing Calamansi may be applied to the face and body in the morning and at

night. Twice daily application of these ingredients allows for smoother skin, due to the acidic

nature of Calamansi, which helps to overturn dead skin cells. Sometimes, extra sunscreen with

UVA.UVB protection should be used, to counteract the extra sensitivity of the complexion after

exposure to fruit acids, such as Calamansi.(Hayes, 2011)

This Filipino beauty secret is found in the splendid tropical regions of the Southeast

Asian island; Calamansi trees grow in abundance, offering Filipino men and women easy

access to this healthful extract (Hayes, 2011)

The Calamansi Plant in the Philippines is as ubiquitous as the Calamansi Plant is

common. The Calamansi Plant is often used to season dishes, preserve food, remove stains in

clothing, deodorize and whiten the skin, and of course, make into juice. A prevalent ingredient

found in most local commercial products, The Calamansi Plant is definitive and preferred in

daily cleansing supplies. Ironically, what is not as common and certainly not ubiquitous is the

Calamansi (Citrofortunella microcarpa, Bunge 1832) Essential Oil. (Hill, 2015,)

kitchens and homes, and has evolved to a hundred uses in a household. The Calamansi can be

used as a bleaching agent on laundry, a skin whitener, deodorizer, follicle and hair growth

stimulator and many more. The essential oil comes from the rind of the Calamansi which are

extracted either by centrifuge or steam distillation.(Lorenzo, 2012)

The scent of Calamansi has been proven to help alleviate depression and anxiety. The

Calamansi is also among the few aromatics that not just masks bad smell, but completely

neutralizes them, making the essential oil a great additive to cleansing products. (Casa de

Lorenzo, 2012)

The fruit is used in local recipes in northern Indonesia, especially around the North

Sulawesi region. Fish are spritzed with the juice prior to cooking to eliminate the "fishy" smell.

Kuah asang ("sour soup") is a regional clear fish broth made with calamondin juice. (Worren,

2014)

In Florida, the fruit is used in its fully ripe form with a more mature flavor profile than the

unripe version. Tasters note elements of apricot, tangerine, lemon, pineapple and guava. The

peel is so thin, each fruit must be hand snipped from the tree to avoid tearing. The entire fruit

minus the stems and seeds can be used. The Fruit is hand processed and pureed or juiced and

used in various products such as Calamondin cake, coulis and jam. The peels can be

dehydrated and used as gourmet flavoring with salt and sugar. The fruit was popular with

Floridian home bakers in cake form from the 1920s to 1950s. Once women joined the

workforce, the labor-intensive nature of processing the fruit caused the fruit’s diminished use.

Now that commercially made products and fruit are available, the fruit is experiencing a

resurgence. (Potonia, 2014)

of lemonade or limeade, as mentioned above, and, left a bit sour, lemonade cuts thirst with the

distinctive calamondin flavor. Also Calamondin can be used on fish and seafood, or wherever

any other sour citrus would be used.(Worren, 2014)

Some say that the plant is native to China and is one of the most cold hardy citrus. The

fruits’ cultivation has spread throughout Southeast Asia, India, Hawaii, the West Indies, and

Central and North America.(Worren, 2014)

In North America, Citrofortunella microcarp , Bunge 1832 is grown primarily as

an Ornamental plant in gardens, and in pots and container gardens on terraces and patios. The

plant is especially attractive when the fruits are present.(Worren, 2014)

The plant is frost sensitive and therefore limited outdoors to frost-free climates (such as

Florida, coastal California, South Texas, and Hawaii in the United States). Potted plants are

brought into a greenhouse, conservatory, or indoors as a houseplant during the winter periods

in regions with cooler climates. (Worren, 2014)

However, in the native homeland in Southeast Asia, the Calamondin is easy to cultivate.

The plant grows well in cool and elevated areas and in sandy soils rich in organic matter.

Waterlogged areas are not suitable for cultivation because calamansi plants cannot tolerate too

much moisture. Calamondin can be propagated by seeds using the vegetative parts. To

produce big, luscious fruits, applying fertilizer, such as Ammonium sulfate or urea, around each

tree one month after planting is essential. The trees will start to bear fruit one or two years after

planting. Trees have an average life span of five years. Florida natives, however, report growing

Calamondin trees with life spans in excess of 20 years in some cases, throughout 1950 to the

70's, and throughout the 70's to 90's. So the actual lifespan is an open issue and may depend
on climatic factors, whether the plant is potted or free grown as a citrus yard tree. (Morton, J,

2009)

The Calamondin bears a small citrus fruit that is used to flavor foods and drinks. Despite

the outer appearance and its aroma, the taste of the fruit itself is quite sour, although the peel is

sweet. Eating a whole fruit has a surprise with the combination of sweet and sour.

Calamondin marmalade can be made in the same way as orange marmalade. Like the citrus

fruits, the calamondin is high in vitamin C. (Jacob, 2014)

The fruit can be frozen whole and used as ice cubes in beverages such as tea, soft

drinks, water, and cocktails. The juice can be used in place of that of the common Persian lime

(also called Bears lime). The juice is extracted by crushing the whole fruit, and makes a flavorful

drink similar to lemonade. A liqueur can be made from the whole fruits, in combination

with vodka and sugar.( Lyle, 2006)

Calamansi is a smooth and slightly spiny plant, growing to a height of 3 to 5 meters.

Leaflets are elliptic to oblong-elliptic, 4 to 8 centimeters long. Petioles are very narrowly or

scarcely winged, about 1 centimeter long. Flowers are axillary, solitary, rarely in pairs, white,

and short-stalked. Fruit is yellow when ripe, nearly spherical, 2 to 3.5 centimeters diameter, 6- to

7-celled, and thin-skinned. The skin or peel is green to yellowish green or yellow, loosely

adhering to the flesh. The flesh contains a few light orange seeds. (Hades, 2014)

The Calamansi (citrofortunella microcarpa, Bunge 1832) is considered as one of the

major fruit crops in the Philippines, which is indigenous to the country. This plant is

characterized by wing-like appendages on the leaf stalks and white or purplish flowers. The fruit

has either a spongy or leathery rind with a juicy pulp that is divided into sections. Calamansi is

said to be a good source of vitamin C that that fruit has been processed into syrups, juices,
concentrates, and purees. The juice is used as flavorings or as an additive in various food

preparations to enhance iron absorption. The juice can also be a preserve ingredient for sweet

pickles or marmalade. Calamansi is easy to cultivate. The plant grows well in cool and elevated

areas and in sandy soils rich in organic matter. Waterlogged areas are not suitable for

cultivation because calamansi plants cannot tolerate too much moisture. (Potonia, 2014)

Citrus is a common term and genus of flowering plants in the rue family, Rutaceae. These

plants are large shrubs or small trees, reaching 5-15 m tall, with spiny shoots and alternately

arranged evergreen leaves with an entire margin. The flowers are solitary or in small corymbs,

each flower 2-4 cm diameter, with five (rarely four) white petals and numerous stamens; they

are often very strongly scented. (Rika, 2015)

Citrus fruits are notable for their fragrance. The juice contains a high quality of citric acid

giving them their characteristic sharp flavor. The genius is commercially important as many

species are cultivated for their fruit, which is eaten fresh pressed for juice, or preserved in

marmalades and pickles. ( Muljadi,2014)

Hand sanitizers have a form of alcohol, such as ethyl alcohol, as an active ingredient. It

works as an antiseptic. Other ingredients may include water, fragrance, and glycerin.

Alcohol-based hand sanitizers are clearly a very useful and important method to prevent

most bacterial and viral infections, with rare exceptions. (Doheny, 2013)

A typical, store-bought hand sanitizer contains the active ingredients isopropanol or

ethanol; these are both forms of alcohol. These types of alcohol have the ability to kill many

germs on contact, without damaging the skin’s tissue. Isopropanol and ethanol kill germs by

dissolving their constituent proteins, causing the germs to die.

the hands, increased convenience, quick-acting, can cause less irritation than soap and may

improve the skin’s condition. Using a hand sanitizer can protect your health and help prevent

the spread of bacteria. (Eden, 2015)

The invention of hand sanitizer was from Lupe Hernandez of Bakersfield, California. In

1996, Lupe Hernandez called an invention hotline off of the TV, and ordered a package for an

idea that Lupe had on how Lupe would create system in hospitals, to where all sinks would have

an alcohol solutions, at the bottom of the sink connected to the pipe, outgoing water reaching

the hands with cleaning solution and a sanitizer gel making the system easier for hospital staff

and quicker to get patients. This idea came to mind because Lupe was attending school to

become a RN. (Chelsea, 2014)

These normal microflora such as Staphylococcus aureus resident in the human skin can therefore be

passed from one individual to another. An easy way by which microbes that are not resident in the

hands are contacted is by contact with surfaces such as doorknobs or handles, toilet handles and taps in

restrooms. Microbes carried on the human skin are of two types i.e. resident and transient.The resident

types are the normal flora while the transient are contacted from the environment. Teaching

appropriate hand hygienic practice have numerous benefits in various environments such as learning

institution including creches, kidergatens, high schools and college as well as promote general

wellness.(Sandra, 2015)

In a study entitled ”Antimicrobial activity of essential oil and various extracts of fruits of

greater cardamom” conducted by Agnihotri S and Wakode S, Greater cardamom (Amomum

subulatum Roxb. Zingiberaceae) commonly known as "Bari ilaichi" is a well-known plant used in

Ayurvedic and Unani medicine. Cardamom has been used for the treatment of various diseases
and disorders like gastric ulcer. Therefore antimicrobial activity of petroleum ether, methanol

and aqueous extracts from leaves and roots, essential oil and isolated vasicine from A. vasica

were tested against various microorganisms. Antimicrobial activity was done by disc diffusion

method. The zone of inhibition observed was compared with that of standard drugs,

ciprofloxacin and fluconazole. Minimum inhibitory concentration was determined against

microorganisms used in the study. The results of this study reveal that methanol extract of fruits

of A. subulatum shows remarkable antimicrobial activity against Escherichia coli whereas in

case of other microorganisms used the extract was found inferior to the standard drug used.

Methanol extract of rind showed good antimicrobial activity against Staphylococcus aureus. The

essential oil isolated was found effective against majority of microorganisms used viz. Bacillus

pumilus, Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa,

Saccharomyces cerevisiae. (Agnihotri S, Wakode S, 2010)

Justification of the Present Study

This study specifically aims to produce an antibacterial hand sanitizer from Calamansi

pure extract. Calamansi is a type of citrus according to the past studies which enabled former

researchers to produce hand sanitizers from various citrus fruits, including calamansi. Utilization

of the fruits’ rinds were implemented, while the researchers will utilize the calamansi pure

extract.The current study focuses on the calamansi pure extract's antibacterial properties in

order to utilize the pure extract as an antibacterial hand sanitizer.