Effectivity of “blumea balsamifera” (Sambong) Leaves as an Alternative

Antibacterial Soap against Staphylococcus Aureus

A Research Proposal Presented to the Faculty

of Senior High School of
DMMC Institute of Health Sciences

In Partial Fulfilment of the Requirements for the completion of Research 2

Senior High School Students
STEM-Fleming

By
Trinidad, Rexel A.

Evangelista, Angelica Jean D.

Adona, Jermaine D.

Sobrepeña, Hannah V.

2021
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Chapter I

THE PROBLEM AND ITS BACKGROUND

This chapter contains the following parts: Introduction; Statement of the Problem;

Theoretical and Conceptual Framework; Hypothesis; Scope and Limitations of the Study;

Significance of the Study; and Definition of Terms.

Blumea balsamifera (sambong) leaves are a kind of leaf that provides numerous health

benefits. Its primary uses are as a diuretic and to treat common cold symptoms. Sambong is a

diuretic herb that is used to treat urolithiasis (urinary tract or kidney stones) and urinary tract

infections, and thus lowers blood pressure.

Sambong leaves have many advantages. Blumea balsamifera (sambong) leaves are used

in Thailand, China, and many Asian countries for the treatment of septic wounds and other

infections. Since it has antimicrobial properties, it can also be used as antibacterial soap.

This study aims to make an alternative antibacterial soap from blumea balsamifera, which

is normally seen in backyards. The researchers had an idea and make these leaves more

convenient and explore other diseases such as the known bacteria— staphylococcus aureus.

Furthermore, some of the content of a commercial soap is fully made of chemicals that can cause

destructions to our fine skin. However, organic substances such as herbal plants contain natural

effects and appropriate to use for adults and even teens or kids. In order to help and sustain the

health of our environment, the more we work together and make plants that will give us an

alternative product that reduces the contamination of the bacteria, we will have a safer

community and low-budget products at the same time we are creating a change into this world.
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In addition, soap is one of the regular needs of almost all people worldwide, generally use as a

cleansing agent and also for other personal hygiene purposes, in which some of them are really

expensive yet less effective. A bar of profitable bar soap in the market is usually the most

frequently used nowadays with a softening and glowing effect on the skin. A good quality soap

has a great impact on nourishing our skin and buying the inappropriate soap could lead to any

harmful result not just to ourselves but also to our family that uses the same products. Some of

the soap ingredients like fragrance are one of the kinds that might cause a trigger on our skin.

This material was evaluated for genotoxicity, repeated dose toxicity, developmental toxicity,

reproductive toxicity, local respiratory toxicity, phototoxicity. (AM Api, D Belsito, S Bhatia,

2015)

The aim of this study is to see how efficient Blumea balsamifera (sambong) leaves are as

an organic and alternative antibacterial soap pertaining to a fast recovery and preventing the

rapid spread of bacteria in a cut or wound, with the least amount of chemical in comparison to

any leading antibacterial soap that is currently available in the market. Furthermore, it will

inspire and encourage Filipinos, especially young people, to develop any biodegradable and/or

non-biodegradable materials that would otherwise be thrown away and generate into new items

and products. This study would motivate, encourage, and also build up the youth to start their

own business at a very young age by selling products they created themselves. It will help them

to earn money together with the lessons, they will be going to learn on how to be more

independent and productive.

This research will look into the effectiveness of Blumea balsamifera (sambong) leaves as

an organic and alternative antibacterial soap. In addition, if the researchers' antimicrobial soap is
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already in use, the effects and potential outcomes will be evaluated in this study. Lastly, the

information obtained would also be beneficial to the proposed product's target market.

PURPOSE STATEMENT

The purpose of this experimental study is to determine the effectiveness of Blumea

Balsamifera (Sambong) extract as an alternative antibacterial bar soap. In this statement, the

success of the antibacterial and as a remedy of the sambong extract will be defined as the main

ingredient that has the ability to cure the staphylococcus disease. Aqueous and alcoholic extracts

of sambong were tested against all 5 resistant bacteria to a maximum concentration of 50,000

ug/mL due to the thickness of the extract. The MICs ranged from 12.5 to >50 mg/mL, showing

some antibacterial activity for the pathogens tested. S. pneumoniae had the lowest MIC using the

alcoholic sambong extract. Leaves of Blumea balsamifera (sambong) are used in traditional Thai

and Chinese medicine for the treatment of septic wounds and other infections. In this study, the

essential oil, hexane, dichloromethane, and methanol extracts of these leaves were evaluated for

antibacterial and antifungal activities using the disc diffusion assay and agar microdilution

method. This plant's active constituents could potentially be developed for use in the treatment

and/or prevention of microbial disease.

STATEMENT OF THE PROBLEM

This study will be conducted to determine the Effectivity of “blumea balsamifera”

(Sambong) leaves as an alternative antibacterial soap against staphylococcus aureus.

The following questions need to be answered:

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1. What is the level of effectiveness of “blumea balsamifera” (Sambong) leaves as an

alternative antibacterial soap against staphylococcus aureus using the following

procedures?

a. 100%

b. 75%

c. 50%

d. 25%

2. Is there any significant difference in the level of effectiveness of “blumea balsamifera”

(Sambong) leaves as an organic antibacterial soap compared to economically produced

soap in the market?

3. How many results of the investigation become beneficial after the study has been proven?

THEORETICAL FRAMEWORK

Staphylococcus aureus is a Gram-positive bacterium and causative agent of a wide range

of infectious diseases such as skin infections, bacteremia, endocarditis, pneumonia, and food

poisoning. The organism was originally a leading nosocomial pathogen and afterward,

epidemiologically distinct clones emerged in community settings. S. aureus cells are Gram-

positive and appear spherical. They are often in clusters resembling a bunch of grapes when

observed under a light microscope after Gram staining. The name ‘Staphylococcus’ was derived

from Greek, meaning a bunch of grapes (staphyle) and berry (Kokkos). The term ‘aureus’ is

derived from Latin, which refers to the color of gold. (proposed by Arumugam Gnanamani,

Periasamy Hariharan and Maneesh Paul- Satyaseela, 2017).

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Staphylococcus aureus is recognized as one of the major foodborne pathogens in fresh

and ready-to-eat products and responsible for various infections around the world (Diep et al.,

2006). It could grow at a temperature between 15°C and 45°C and at NaCl concentrations as

high as 15% (Behling et al., 2010). This bacterium multiplies quickly at room temperature to

produce toxins that cause illness. Naturally, the distribution of S. aureus was ubiquity in the

world, but the most important infection source of S. aureus was food. Every year, S. aureus is

involved in about 241,000 illnesses of foodborne disease in the USA (Scallan et al., 2011; Wu et

al., 2018). In 2013, there were 12.5% of foodborne bacterial outbreaks are caused by S. aureus in

China, which showed the third most frequently pathogen after Vibrio parahaemolyticus (27.8%)

and Salmonella (23.1%) (Wei-Wei et al., 2018).

CONCEPTUAL FRAMEWORK

INPUT PROCESS OUTPUT

Observation and monitoring of

effects and reactions in the Analyzing the data gathered Recommendation of the

sambong leaves relating to using statistical tools and homemade antibacterial

recovery and preventing the assessing results by soap against

rapid spread of bacteria in a categorizing results and Staphylococcus Aureus that

cut or wound after the reactions for every common is made of sambong leaves

application of the homemade kind of sambong leaves with positive results and

antibacterial soap against tested. reaction and as an

Staphylococcus Aureus made alternative for store-bought

of sambong leaves. antibacterial soap.

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Figure 1. The Conceptual Framework of the Study

The researchers use the standard IPO format to further explain our research: This model

of Morris and Gouran (1973-1975) encloses the input that contains blumea balsamifera

(sambong) extract as the primary ingredient to create antibacterial soap that could be a remedy

for restraining the bacteria to grow.

The process examines the antibacterial soap of the sambong extract as we conduct

different samples and have the trial variations in which we will identify the probable efficacy,

content, measures, and amount of every ingredients needed on the experimental study before

creating the product.

The output includes the study's findings for recommending homemade antibacterial soap

as an alternative to store-bought antibacterial soap based on data about its efficacy on certain

types of wounds and cuts.

NULL HYPOTHESIS

Ho: There is no significant difference in the effectiveness of the “blumea balsamifera”

(Sambong) leaves as an antibacterial soap compared to the soap economically produced soap in

the market.

ASSUMPTION OF THE STUDY

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The research study is anchored on the assumption that the homemade blumea balsamifera

(Sambong) Leaves as an Alternative Antibacterial Soap can treat various wounds primarily the

Staphylococcus Aureus. However, if it is not very effective in curing Staphylococcus Aureus, the

researchers are assuming that it can still be beneficial when it comes to treating sorts of wounds

and cuts particularly to our skin.

SCOPE AND DELIMITATION OF THE STUDY

The study covers the overall result of the alternative antibacterial soap and its use in the

community and also to the environment by using the extract as the main content of the product.

This will be tested in various trials using the extract as soap. Hence, the commercial products of

antibacterial soap will be the guide when comparing the effectiveness. It delimits that the study

might not be able to make the result effective in general; a person with other related diseases

may not be able to apply this product.

SIGNIFICANCE OF THE STUDY

Through this research, others will be reminded that this type of leaf can be used to

make antibacterial soap. When it comes to washing wounds or cuts, one will be aware of the

potential and importance of using organic products. It will also help those who are concerned

about their hygiene and environmentalists, as this study aims to create all-natural

antibacterial soap. The children who suffer from cuts and wounds as a result of their

playfulness and carelessness are also among those who will benefit from this research.
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Furthermore, this would raise health consciousness in order to protect children from

chemical-based cleaning products. In conclusion, this research will raise awareness about the

importance of recycling products that would otherwise be discarded.

This study will benefit the following:

The Students - This research is significant, first and foremost to the students. They are the

subject of this study after all. Knowing that the effects of making an antibacterial soap will

encourage them to start a business on their own and earn income for themselves together

with the lessons they will be going to learn on how to be more independent and productive at

a very young age.

The Community - The result of this study will give them information that sambong leaves

can be used as antibacterial soap to save money and minimize their waste.

The Researchers - Researchers are also still part of the education system since they are the

people behind this academic study, they can propose how to manage to develop sambong

leaves into antibacterial soap and generate a business with the use of it. If they can manage to

prove this study, this will help the students to be more independent and productive because of

the understanding that the information obtained by the researcher.

Future Researchers - The study will give them the research format and will serve as their

guide and reference for their own research paper.

DEFINITION OF TERMS
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There are some terms needed to be defined in this study for clarify and further

understanding. The following terms were defined by the researchers to understand the research

paper.

Antimicrobial. Referring to an agent or mechanism that kills or inhibits the growth or

reproduction of microbes.

Bacteremia. The presence of live bacteria in the bloodstream.

Blumea balsamifera. A medicinal herb, is widely distributed in Eastern and Southeastern Asia.

It has been used in folk medicine for its many health benefits that include the treatment of septic

wounds, respiratory infections, stomach pains and kidney stones.

Diuretic. Are medications designed to increase the amount of water and salt expelled from the

body as urine.

Endocarditis. Is an infection caused by bacteria that enter the bloodstream and settle in the heart

lining, a heart valve or a blood vessel.

Genotoxicity. Refers to processes that alter the structure, information content, or segregation of

DNA and that are not necessarily associated with mutagenicity.

Hemostasis. Is a process to prevent and stop bleeding, meaning to keep blood within a damaged

blood vessel.

Inflammation. A localized physical condition in which part of the body becomes reddened,

swollen, hot, and often painful, especially as a reaction to injury or infection.

Septic Wound. A wound that has become infected.

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Skin Infections. Is a condition where bacteria or other germs enter the skin through a wound and

spread, causing pain, swelling, and discoloration.

Staphylococcus aureus. Is a gram-positive bacteria that cause a wide variety of clinical

diseases.

Synthetic Drugs. Are created using man-made chemicals rather than natural ingredients.

Urinary tract infection (UTI). Is an infection in any part of your urinary system — your

kidneys, ureters, bladder and urethra.

Urolithiasis. The process of forming stones in the kidney, bladder, and/or urethra (urinary tract).

Wound Healing. Is a complex and dynamic process of replacing devitalized and missing cellular

structures and tissue layers.

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Chapter II

REVIEW OF RELATED LITERATURE AND STUDIES

The study's analysis focuses on the history of blumea balsamifera and its antibacterial

properties. The internet was used as a reference source to obtain the relevant data and details for

the analysis.

A. Foreign Literature

Blumea balsamifera—A Phytochemical and Pharmacological Review

According to (Yuxin Pang, Dan Wang, Fulai Yu, 2014) Nowadays, herbal medicines are

widely consumed and their sales have been rising significantly all over the world. According to

the reports of the World Health Organization (WHO), to treat diseases over 80% of the

populations in developing countries mainly rely on herbs, which are considered to be safer and

more effective than synthetic drugs.

Blumea balsamifera (L.) DC. (Asteraceae), also known as sambong, has been used as

medicine for thousands of years in Southeast Asia countries, such as China, Malaysia, Thailand,

Vietnam, and Philippines. Sambong is the most important member of the genus Blumea and is an
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indigenous herb of tropical and subtropical Asia, especially in China. This plant grows on forest

edges, under forests, river beds, valleys and grasses

The whole plant or its leaves were used as a crude Chinese traditional medicinal material

to treat eczema, dermatitis, beriberi, lumbago, menorrhagia, rheumatism, skin injury, and as an

insecticide. Bing Pian and Aipian are two important traditional Chinese medicines (TCMs)

extracted from plants and have been used as one in prescriptions for centuries in China.

The survey and summary of the extensive studies revealed that B. balsamifera was an

essential and valuable medicinal plant used for folk treatments such as treating eczema,

dermatitis, beriberi, lumbago, menorrhagia, rheumatism, skin injury, or used as insecticide. As a

traditional medicine, the biological and pharmacological studies of the plant materials, crude

extracts, and isolated chemical constituents of B. balsamifera offered experimental and scientific

proofs for its various traditional uses. The pharmacological studies focused on studying the anti-

microbial and anti-inflammatory effects, antiplasmodial effects, platelet aggregation, wound

healing, and disease and insect resistant activities, all of which confirmed the plant’s traditional

uses.

Effect of volatile oil from blumea balsamifera (l.) Dc. Leaves on wound healing in mice

According to (Yuxin Pang, Dan Wang, Xuan Hu, Hui Wang, Wanjin Fu, Zuowang Fan,

Xiaolu Chen, Fulai Yu, 2014) Traditional medicine practitioners have described the therapeutic

efficacies of many traditional and indigenous plants against diseases. Natural products that are

safe, and possess physiological properties are excellent sources of new therapeutics for the

treatment of conditions like mechanical damage of the skin. Some researchers, therefore, have
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shifted their focus to the potential wound healing properties of plants. Wound healing is a

process of restoring damaged cells and tissues. The phases of wound healing occur in a precise

and regulated order. Firstly, it involves hemostasis and inflammation; secondly, mesenchymal

cells differentiate, proliferate, and migrate to the wound site; thirdly, angiogenesis and epithelial

deposition at the wound surface site. Finally, synthesis, cross-linking, and alignment of collagen

are required to provides structural strength to the wound healing tissue. The wound also

undergoes physical contraction, which might be mediated by contractile fibroblasts.

Neuropeptide Sub-stance P (SP) is a pro-inflammatory neuropeptide, and modulates

inflammatory responses of skin wounds. SP also promotes the synthesis and metabolism of fibro-

blast and increases accumulation of collagen in the proliferative phase of mesenchymal cell

growth and dynamics. In addition, SP is an important medium in the process of wound repair and

scar healing.

Ainaxiang (Herba Blumeae Balsamiferae), also named Sambong in some tropical

countries, is an herb with a rich constitution of essential oils that are used in Traditional Chinese

Medicine (TCM). In China, Blumea Balsamifera (L.) DC. normally grows in Hainan and

Guizhou. Its leaves have been used for healing many conditions including eczema, dermatitis,

skin injury, skin bruises, beriberi, lumbago, menorrhagia, rheumatism, and some other diseases.

Recently, the extracts of the leaves have been verified to display physiological activities on

plasmin-inhibitory, anti-fungal, free radical scavenging, and anti-obesity functions.

In this study, BB oil and BB oil demonstrated a significant increase in the hydroxyproline

content of the granulated tissue, indicating an increase of collagen turnover. Both these two

diluted BB oils stimulated wound healing more potently and more effectively than did the others.

These findings implied that BB oil BB oil promoted wound healing by influencing the
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proliferative and remodeling phases of wound healing, and do so via the pathways of angio-

genesis, collagen deposition, granular tissue formation epithelial deposition, and wound

contraction. This result was also relevant in the context of other traditional medicinal plants.

Topical applications of many traditional medicinal plants for skin wounds have been proved to

have certain positive effects. These findings made these folk herbs potential skin wound healing

agents.

Antimicrobial activity of blumea balsamifera (lin.) Dc. Extracts and essential oil

According to (Uthai Sakee et al. Nat Prod Res., 2011) Leaves of Blumea balsamifera

(Lin.) DC. are used in traditional Thai and Chinese medicine for the treatment of septic wounds

and other infections. In this study, the essential oil, hexane, dichloromethane and methanol

extracts of these leaves were evaluated for antibacterial and antifungal activities using the disc

diffusion assay and agar microdilution method. The essential oil was the most potent, with a

minimum inhibitory concentration (MIC) of 150 µg mL−1 against Bacillus cereus and an MIC of

1.2 mg mL−1 against Staphylococcus aureus and Candida albicans. Activity was also detected

from the hexane extract against Enterobacter cloacae and S. aureus. Minimum bactericidal and

fungicidal concentrations were typically equal to or two-fold higher than the MICs for both

extracts, indicating microbicidal activity. The present data show that B. balsamifera extracts have

activity against various infectious and toxin-producing microorganisms. This plant's active
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constituents could potentially be developed for use in the treatment and/or prevention of

microbial disease.

Blumea balsamifera Oil for the Acceleration of Healing of Burn Injuries

According to (Zuo-Wang Fan, Yu-Xin Pang, Kai Wang, 2015) Blumea balsamifera oil

(BBO) is a main extract obtained from Blumea balsamifera (L.) DC (Ainaxiang) leaves, which

are widely used as a traditional medicine by the Miao and Li Nations to promote skin trauma or

burn injury healing. This study was initiated to investigate the healing efficacy in deep second-

degree burn model in rats. The rats were treated by BBO for 21 consecutive days. The rate of

healing, scabs dropped time and re-epithelialization time were observed every three days for 21

days after burn injury. The samples were collected from different treated rats by sacrificing the

animals on the 1st, 2nd, 5th, 9th, 14th, and 21st day post-burn creation. Then, the water content

of burn tissue was measured. Plasma interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-

α) levels were evaluated, and the tissue expressions of basic fibroblast growth factor (bFGF),

vascular endothelial growth factor (VEGF), and transforming growth factor-beta (TGF-β) were

determined along with skin histopathology. The results showed that the water content of tissue

was significantly reduced, the scabs dropped time shortened, and healing accelerated after

treatment with BBO in the burn injury rats. Furthermore, the expressions of growth factors were

significantly increased in the tissue; however, the levels of inflammatory factors on plasma

decreased. This study confirms the efficacy of BBO consumption on burn injuries.

Phytochemical composition and health properties of Sembung plant (Blumea balsamifera)

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According to (Gede Widhiantara and Made Jawi, 2021) Indonesia’s mindset has been

focusing on the use of natural medicines, food, and healing practices widely recognized by the

nation’s culture. Traditional medicines and herbs used in traditional medicine can often lead to

the discovery of drugs against certain diseases. The aim of this review was to study evidence-

based data on the importance of Sembung plant, Blumea balsamifera, as a potential traditional

medicine. The distribution, ethnopharmacology, secondary metabolites, and bioactivity against

several diseases are focused in this review. Information and research related to Sembung plant

were searched using the terms “B. balsamifera,”“phytochemicals,” and “pharmacological

activity” on ResearchGate, Google Scholar, Science Direct, PubMed, and scientific information-

based databases up to 2020. Several ethnomedical articles recommend B. balsamifera for the

treatment of sinusitis, colic pain, cough, kidney stones, flu, or as a diuretic. This knowledge has

already been applied in several countries in Southeast Asia. B. balsamifera has been reported to

contain several phytochemicals both volatile (terpenoids, fatty acids, phenols, alcohol,

aldehydes, ethers, ketones, pyridines, furans, and alkanes) and non-volatile (flavonoids,

flavanones, and chalcones). Extracts and phytochemicals of B. balsamifera contain several

biological capacities such as antioxidant, antimicrobial, antifungal, anti-inflammatory,

hypolipidemic, antiinfertility, hepatoprotective activity, antidiabetic, gastroprotective, antitumor,

anticancer, and immunomodulatory agent against Coronavirus disease-19 infection. This review

provides essential data for the potential application of B. balsamifera as a nutraceutical or in

future medicinal preparations.

Biological Activities and Chemical Compositions of Volatile Oil and Essential Oil from the

Leaves of Blumea balsamifera

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According to (Yuan-Hui Wang, Xiao-Yu Yu, 2018) A modified hydrodistillation was

designed to isolate essential oil (EO) from the leaves of Blumea balsamifera (L.) DC. cultivated

in Luodian County, China. Meanwhile, hydrodistillation-solvent extraction (HDSE) and

simultaneous distillation and extraction (SDE) were used to collect volatile oils of Luodian B.

balsamifera. Chemical compositions, antioxidant and antimicrobial activities of volatile oil and

EO of Luodian B. balsamifera were investigated. GC-MS and GC-FID analysis showed that

caryophyllene, xanthoxylin, γ-eudesmol, α-cubenene were the main compounds of EO. In the

tests of antioxidant and antimicrobial capabilities, the antioxidant activity of EO was better than

the other two volatile oils, and EO had high antioxidant activity in DPPH free radical scavenging

test (IC50: 28.22 g/L), β-carotene bleaching inhibition test (IC50: 3.27 g/L), and thiobarbituric

acid reactive species assay. EO exhibited favourable antifungal activity (MIC: 62.5–250 μg/mL),

and showed antibacterial activity against Staphylococcus aureus (MIC: 2000 μg/ mL). Essential

oil of B. balsamifera leaves has potential as antiseptic and antioxidant.

Effects and Mechanisms of Total Flavonoids from Blumea balsamifera (L.) DC. on Skin

Wound in Rats

According to (Lingliang Guan, Yingbo Zhang, Fulai Yu, 2017) Chinese herbal medicine

(CHM) evolved through thousands of years of practice and was popular not only among the

Chinese population, but also most countries in the world. Blumea balsamifera (L.) DC. as a

traditional treatment for wound healing in Li Nationality Medicine has a long history of nearly

2000 years. This study was to evaluate the effects of total flavonoids from Blumea balsamifera

(L.) DC. on skin excisional wound on the back of Sprague-Dawley rats, reveal its chemical

constitution, and postulate its action mechanism. The rats were divided into five groups and the

model groups were treated with 30% glycerol, the positive control groups with Jing Wan Hong
 19

(JWH) ointment, and three treatment groups with high dose (2.52 g·kg-1), medium dose (1.26

g·kg-1), and low dose (0.63 g·kg-1) of total flavonoids from B. balsamifera. During 10

consecutive days of treatment, the therapeutic effects of rates were evaluated. On day 1, day 3,

day 5, day 7, and day 10 after treatment, skin samples were taken from all the rats for further

study. Significant increases of granulation tissue, fibroblast, and capillary vessel proliferation

were observed at day 7 in the high dose and positive control groups, compared with the model

group, with the method of 4% paraformaldehyde for histopathological examination and

immunofluorescence staining. To reveal the action mechanisms of total flavonoids on wound

healing, the levels of CD68, vascular endothelial growth factor (VEGF), transforming growth

factor-β₁ (TGF-β₁), and hydroxyproline were measured at different days. Results showed that

total flavonoids had significant effects on rat skin excisional wound healing compared with

controls, especially high dose ones (p < 0.05).

Simultaneous Quantitative Determination of Polyphenolic Compounds in Blumea

balsamifera

According to (Daopeng Tan, Zhou Yang, Qianru Zhang, 2020) A high-performance liquid

chromatography method was developed for simultaneous quantification of 18 polyphenolic

compounds from the leaves of Blumea balsamifera, including 17 flavonoids and 1

phenylethanone. The B. balsamifera extraction was separated by a Kromasil C18 column (250 ×

4.6 mm, 5 μm) with a binary gradient mobile phase consisting of acetonitrile and 0.2% aqueous

acetic acid. A photodiode array detector (PDA) was used to record the signals of investigated

constituents. The linearity, sensitivity, stability, precision, and accuracy of the established assay

methods were assessed to meet the requirements of quantitative determination. Samples

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extracted by reflux in 25 mL of 80% methanol for 30 minutes were selected for the extraction

method. The 18 compounds were accurately identified by comparing with the reference

compounds. The purity of each peak was confirmed by the base peak in the mass spectrum. The

contents of 18 compounds in Blumea samples from four different regions were successfully

determined. The results also showed that 3,3',5,7-tetrahydroxy-4'-methoxyflavanone was the

most abundant constituent, which could be used as a potential chemical marker for quality

control of B. balsamifera and Chinese patent medications containing B. balsamifera herb.

Antibacterial effect of Blumea balsamifera DC. essential oil against Haemophilus parasuis

According to (Changliang He, Peiyi Yang, Lu Wang, Xiaolin Jiang, 2020) Haemophilus

parasuis (H. parasuis), the cause of the Glasser’s disease, is a potentially pathogenic gram-

negative organism that colonizes the upper respiratory tract of pigs. The extraction of Blumea

balsamifera DC., as a traditional Chinese herb, has shown great bacteriostatic effect against

several common bacteria. To study the antibacterial effect on H. parasuis in vitro, this study

evaluated the minimum inhibitory concentration (MIC) and minimum bactericidal concentration

(MBC) of Blumea balsamifera DC. essential oil (BBO) as well as morphological changes in H.

parasuis treated with it. Furthermore, changes in expression of total protein and key virulence

factors were also assessed. Results showed that the MIC and MBC were 0.625 and 1.25 μg/mL,

respectively. As the concentration of BBO increased, the growth curve inhibition became

stronger. H. parasuis cells were damaged severely after treatment with BBO for 4 h,

demonstrating plasmolysis and enlarged vacuoles, along with broken cell walls and membranes.

Total protein and virulence factor expression in H. parasuis was significantly downregulated by
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BBO. Taken together, these results indicated a substantial antibacterial effect of BBO on H.

parasuis.

Bioactive Sambong oil-loaded electrospun cellulose acetate nanofibers: Preparation,

characterization, and in-vitro biocompatibility

According to (Azeem Ullaha, Yusuke Saitoa, Sana UlahBlumea balsamifera oil loaded

cellulose acetate nanofiber mats were prepared by electrospinning. The inclusion of blumea oil

increased the nanofiber diameter. FTIR spectra confirm the addition of blumea oil in the

nanofiber mats. The XRD pattern suggests that the inclusion of blumea oil has caused a

misalignment in the polymer chains of the cellulose acetate. Thus, a decrease in the tensile

strength was observed for the blumea oil loaded nanofibers. The increase in fiber diameter

causes a reduction in the porosity of the nanofiber mats. The blumea oil loaded nanofiber mats

showed antibacterial efficacy against Escherichia coli and Staphylococcus aureus. The blumea

oil showed antioxidant abilities against the DPPH solution. MVTR of the neat and blumea oil

loaded nanofiber mats was in the range of 2450–1750 g/m2/day, which is adequate for the

transport of air and moisture from the wound surface. Blumea oil loaded mats showed good cell

viability ~92% for NIH 3T3 cells in more extended periods of incubation. A biphasic release

profile was obtained, and the release followed the first-order kinetics depending upon the highest

value of the coefficient of correlation R 2 (88.6%).

Kajian Etnobotani Pakundalang (Blumea balsamifera (L.) DC.)

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According to (Wardah Wardah, Emma Sri Kuncari, 2020) Banggai Kepulauan Regency

(Bangkep) is a small and outermost island in Indonesia, has problems with slow access to and

from outside due to inadequate transportation and telecommunications facilities. Therefore,

people's dependence on natural resources around them, especially medicinal plants, is essential.

The method used in this study was selected interviews with village heads, traditional leaders,

shamans, traditional healers, and villagers who use plants as medicinal ingredients. The results of

study carried out in several villages in Banggai Kepulauan Regency showed that the community

still use plant resources that are growing around to treat various ailments. One type of plant that

is widely used as a medicinal ingredient is pakundalang (Blumea balsamifera (L.) DC.) which

belongs to the Asteraceae family. Pakundalang is an alternative solution for community health

independence to overcome various ailments including postpartum treatment, fever, flu, pain

during menstruation, hemorrhoids, internal wounds, skin pain, cough, high blood pressure,

diarrhea, rheumatism, stomach pain, diabetes, shortness of breath, heartburn, and malaria. Data

and information regarding the ethnobotany of pakundalang are discussed in this article.

Utilization of Asteraceae family as medicinal plant by local community of hutan pinus

Jantho nature reserve, Aceh Besar

According to (Liza Fadhillah, Saida Rasnovi, Zairin Thomy, 2021) The use of medicinal

plants by local communities is an ethnobotany study that links the culture of the community with

the resources of plants that are around it. A study on the utilization of medicinal plants in the

Hutan Pinus Jantho Nature Reserve was conducted with the aim of obtaining information about

the types of medicinal plants used by local communities around Hutan Pinus Jantho Nature

Reserve. This study applies the Participatory Rural Appraisal (PRA) method for interviews
 23

conducted by Purposive Sampling. The parameters measured are; parts of medicinal plants used,

method of processing, method of use, and location of the collection. According to interview, it

found about 10 species of Asteraceae those have medical potential, 3 of them already use by the

local community of CAHPJ, these are Blumea balsamifera L., Chromolaena odorata L., and

Tagetes erecta L., these species could cure the wound, heal the cold, fever, stomachache and

other diseases.

Formulasi Tablet Hisap Ekstrak Etanol Daun Sembung (Blumea Balsamifera (L.) Dc.)

According to (Yaminulloh, Sholeh and, Suprapto, M.Sc., Apt 2017) The treatment of

stomatitis aftosa are caused by Candida albicans is currently drops and mouthwash but differ

with lozenges. Lozenges are an effective treatment for stomatitis aftosa in terms of both

economic and comfort especially lozenges that contain active sembung this. Sembung (Blumea

balsamifera (L.) DC.) has many benefits such as antimicrobial. This study aims to determine the

ethanol extract lozenges formula sembung (Blumea balsamifera (L.) DC.) With a combination of

binder PGA (Pulvis Gummi Arabicum) and sweetener mannitol and determine its activity against

Candida albicans fungus before and after formulation. Extraction done by maceration method,

and then testing the activity of the extract by the method of diffusion wells and the formulation

lozenges comprising five formulas in composition variation binder PGA (Pulvis Gummi

Arabicum) and sweetener mannitol ratio of each formula that is F1 (PGA 2, 5%; Mannitol

17.5%); F2 (PGA 5%; mannitol 15%); F3 (PGA 10%; mannitol 10%); F4 (PGA 15%; Mannitol

5%); F5 (PGA 17.5%; Mannitol 2.5%), manufacture of tablets by wet granulation method.

Testing the physical properties of the tablet include: organoleptic, uniformity of weight,

hardness, friability, disintegration time with the processing of data using ANOVA test, test the
 24

responsiveness sense tablet with data processing using Friedman test. The results of this study

are obtained sembung ethanol extract as much as 50.86 g with a yield of 4.23% w / w, the levels

of 60 ug / mL to provide activity against Candida albicans fungus with inhibition zone of 10 mm,

and the effect is not lost even done formulations. The test results lozenges have met all the

physical properties of tablets required for lozenges, the requirements are: weight uniformity (CV

<5%), hardness (10-20 Kg), friability (<1%), disintegration time (<30 minutes). The best

formula of this research is the formula 3 with mannitol levels and levels of PGA for tablet weight

of 400 mg each at 40 mg / tablet.

A Review: Antioxidant Activities of Sembung Leaves (Blumea Balsamifera (L.) DC)

According to (Septi Dwi Putri, Rusdi and Ridho Asra, 2020) Sembung

(Blumeabalsamifera(L.) DC) is a drug that is widely used for treatment in various countries such

as Indonesia, China, the Philippines and other countries. In Indonesia it has various other names,

in West Sumatra it is called Capo leaf and sembung. In China it is better known as ainiaxing.

Sembung is used as an ancient medical medicine. It has many uses, namely for antioxidants, for

wound healing, antibacterial, anti-malarial, as well as xanthine oxidase inhibitors. Antioxidants

are inhibitors in the oxidation process so that they can protect cells from the dangers of free

radical.

B. Local Literature
 25

Antibacterial
Activity of
Guyabano,
Ulasimang Bato,
Sambong, and
Tsaang Gubat
Leaf Extracts
 26

against Common
Drug-resistant
Bacteria
Antibacterial
Activity of
Guyabano,
Ulasimang Bato,
 27

Sambong, and
Tsaang Gubat
Leaf Extracts
against Common
Drug-resistant
Bacteria
Antibacterial
Activity of
 28

Guyabano,
Ulasimang Bato,
Sambong, and
Tsaang Gubat
Leaf Extracts
against Common
Drug-resistant
Bacteria
 29

Antibacterial
Activity of
Guyabano,
Ulasimang Bato,
Sambong, and
Tsaang Gubat
Leaf Extracts
 30

against Common
Drug-resistant
Bacteria
Antibacterial
Activity of
Guyabano,
Ulasimang Bato,
 31

Sambong, and
Tsaang Gubat
Leaf Extracts
against Common
Drug-resistant
Bacteria
Antibacterial
Activity of
 32

Guyabano,
Ulasimang Bato,
Sambong, and
Tsaang Gubat
Leaf Extracts
against Common
Drug-resistant
Bacteria
 33

Antibacterial
Activity of
Guyabano,
Ulasimang Bato,
Sambong, and
Tsaang Gubat
Leaf Extracts
 34

against Common
Drug-resistant
Bacteria
Antibacterial
Activity of
Guyabano,
Ulasimang Bato,
 35

Sambong, and
Tsaang Gubat
Leaf Extracts
against Common
Drug-resistant
Bacteria
Antibacterial
Activity of
 36

Guyabano,
Ulasimang Bato,
Sambong, and
Tsaang Gubat
Leaf Extracts
against Common
Drug-resistant
Bacteria
Antibacterial activity of guyabano, ulasimang bato, sambong, and tsaan gubat leaf extracts

against common drug- resistant bacteria

 37

According to (Cecilia C. Maramba-Lazarte, Lolita L. Cavinta, and Ma. Carmelita L. Sara,

2020) Bacterial infections remain to be an important cause of morbidity in the Philippines with

respiratory, gastrointestinal and urinary tract infections still being very prevalent. Antibiotics

have been used to treat bacterial infections and are considered one of the most important

discoveries of the modern era. Unfortunately, because of extensive abuse and misuse of

antibiotics both in humans and animals, rates of antimicrobial resistance have been increasing.

According to a WHO Report in 2014, the top 7 common bacteria that cause serious

diseases with widespread resistance include E.coli, S. aureus, H. infuenzae, Shigella, P.

aeruginosa, non-typhoidal Salmonella, Klebsiella, and N. gonorrhea. In the Philippines,

Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii were reported to

have high rates of resistance to conventional antibiotics. What is being feared now is an

occurrence of a post-antibiotic era, wherein present antibiotics are ineffective against common

diseases.

Plants as sources of antimicrobials is an old concept which dates back to traditional

medical systems in numerous cultures. Herbal preparations have been used by traditional healers

as antiseptics, as well as cures for pneumonia, diarrheas, and other bacterial infections. This may

be a logical strategy since plants produce metabolites to protect themselves against microbes in

the environment. Many modern-day medicines and even antibiotics are also nature-derived or

plant-based or plant-derived.

Four commonly used medicinal plants in the Philippines include guyabano, tsaang gubat,

sambong, and ulasimang bato. Guyabano or Anona muricata L (family Annonaceae) is cultivated

throughout the Philippines and is used by Filipino traditional healers for various ailments. The

roots are used for anemia while the leaves are used for cough and colds, indigestion, dysentery,
 38

amebiasis, diarrhea, and various types of pain. In other tropical countries such as Indonesia,

India, and Brazil, guyabano is also commonly used to treat infections such as pneumonia, urinary

tract infection, and skin diseases. Tsaang gubat or Ehretia microphylla Lam. (synonym Carmona

retusa (Vahl) Masam)(family Boraginaceae) or commonly called Philippine tea or Fukien tea, is

found in the Philippines, India, Southern China, Taiwan, and the Malay Peninsula. The

leaves are frequently used by traditional healers as a decoction for various types of

gastrointestinal ailments such as diarrhea, vomiting, abdominal pain as well as for other

indications such as insomnia, fever, cough, colds, and muscle pain. Sambong or Blumea

balsamifera L. (family Compositae/Asteraceae) has been proven as an effective treatment for

kidney stones. In the Philippines, traditional healers recommend using sambong for various

ailments including cough, fever, influenza, dysentery, sore throat, malaria, boils, infected

umbilical cord, sore eyes, and tuberculosis. Ulasimang bato or Peperomia pellucida (L.) Kunth

(family Piperaceae) is also known as pansit-pansitan, sinaw-sinaw, or tangon-tangon. In the

Philippines and West Africa, the whole plant is used as a warm poultice for abscesses and boils.

Other cultures use the leaf decoction to treat urinary tract infections, headache, fever, eczema,

abdominal pains, and convulsions.

Recommended Medicinal Plants as Source of Natural Products

According to (Elseiver B.V., 2018) Many important medicines are natural products or

derived from natural products. More than one third (39.1%) of all Food and Drug Administration

(FDA) approved drugs are of natural origin, and 48.6% of all cancer drugs registered from the
 39

1940s until today are either natural products or derivatives. Natural products are important

sources in the drug discovery process.

Plants in general are rich in antioxidants that are crucial for their survival in the

environment. These antioxidants are often associated with a reduction of health risks that

contribute to conditions such as diabetes. Plant-based traditional medicines are still prevalent

because plants are often inexpensive to prepare, are effective and their use for curing common

ailments results in minimal complications. Thus, plant extracts have become an invaluable

source of candidate compounds for the development of new drugs.

Various methods of treatment exist, including herbal treatment in the Philippines that

uses the medicinal herb Blumea balsamifera.

A very popular Philippine herbal flowering plant used as medicine to treat wounds and

cuts, rheumatism, anti-diarrhea, anti-spams, colds and coughs. It is also used for infected

wounds, respiratory infections and stomach pains. Sambong is very popular among people with

kidney problems because of its diuretic qualities. It can be taken as an early afternoon tea to

maintain a healthy urinary tract. It also helps flush uric acid as well. The Philippine National

Kidney and Transplant Institute recommends taking sambong for patients with renal problems.

Studies noted that it may help to delay dialysis and other kidney problems. Sambong also

possesses antibacterial and antifungal properties.

Blumea balsamifera or sambong is a common medicinal plant found throughout the

Philippines. It is common in open fields, grasslands, and waste areas, at low and medium

altitudes. It also occurs in China, Hainan, Bhutan, Cambodia, Laos, Indonesia, Malaysia,

Thailand and Vietnam.

 40

The effect of varying concentrations of sambong leaves (blumea balsamifera) decoction on

wound healing

According to (Krishna Deo Das, Satyam Prakash and Khushbu Yadav, 2016) Blumeae

balsamiferae was used to treat snake bite injury and skin wounds and itch. It is documented that

external application of the mashed fresh leaves or leaf water washings decoction could treat

traumatic injury, carbuncle and skin pruritus. Natural products that are safe and attain

physiological properties are tremendous sources of new-fangled therapeutics for the treatment of

conditions like mechanical damage of the skin.

As the world rapidly progresses and develops, there seems to be a dominance of

commercially manufactured products and one of which is drugs that man primarily need to cure

ailments and different diseases. The country has always been dependent on highly priced

commercial drugs giving little chance for the utilization of locally grown medicinal herbs and

plants. Therefore, the present study was focussed to determine the effect of a locally grown plant,

Sambong (Blumea balsamifera) on wound healing as an alternative to expensive medicine such

as betadine. The output of the study is the effect of varying concentrations of Sambong leaves

decoction on the wound healing of mice. The findings of this study could be used as reference on

the preparation of Sambong leaves as decoction when used to treat wounds. Thus, this study was

designed to create awareness of the importance of medicinal Sambong plant and provide vital

information regarding its utilization that can be used as an alternative to high-cost manufactured

drugs and preparation in the treatment of wounds.

 41

Xanthine oxidase inhibition of selected Philippine medicinal plants

According to (Karmella L. Apaya and Christine L. Chichioco-Hernandez, 2011) Extracts

from selected species of Philippine plants traditionally used for their anti-inflammatory

properties were examined for their ability to inhibit the action of xanthine oxidase. The degree of

inhibition using Adenanthena payonina, Antegonon leptopus,Blumea balsamifera, Calophyllum

inophyllum, Cassia alata, Cassia fistula, Gliricidia sepium, Michelia alba, Mimosa pudica,

Portulaca olercea, Pogostemon cablin, Solanum tornum, Tinosphora rumphii and Vitex negundo

extracts were determined by measuring the increase in absorbance at 295 nm which is associated

with uric acid formation which is linked to gout. B. balsamifera has the highest percent inhibition

at 79.67% followed by M. pudica with 62.36% inhibition. The xanthine oxidase inhibitory

potential and IC50 values of the extracts are reported.

Medicinal plants used by the Higaonon tribe of Rogongon, Iligan City, Mindanao,

Philippines

According to (Lilybeth F. Olowa, Mark Anthony J. Torres, Eduardo C. Aranico and

Cesar G. Demayo, 2012) This paper presents the ethnobotanical study conducted in Rogongon in

Iligan City, Philippines on the indigenous medicinal plant utilization and healthcare practices

among the Higaonon tribe. Informal interviews, using a semi-structured questionnaire, involving

65 informants (ages 30-78 years old) provided information on 62 plant species of medicinal

value belonging to 34 families and 56 genera. Leaves were the most widely used plant parts

which were commonly prepared by boiling in water (decoction) and administered orally.

Stomach ailments, respiratory diseases, wounds or cuts, and muscle pains or over fatigue in
 42

women were the most common health problems treated by the identified medicinal plants. Some

of the medicinal plants enumerated in the study were also utilized by other indigenous groups in

the Philippines. Such widespread use of these plants by different groups could to a certain extent

be attributed to their efficacy.

Ethnobotanical documentation of medicinal plants used by the Ati tribe in Malay, Aklan,

Philippines

According to (Cecilia Salugta Cordero, Antonio Ligsay, Grecebio Jonathan Duran

Alejandro, 2020) A total of 106 medicinal plant species in 48 families and 95 genera were used

by the Ati to treat 62 diseases across 14 categories. The family Asteraceae was best represented

with 10 species, followed by Lamiaceae with 8, and Apocynaceae and Poaceae with 6 species

each. The most frequently used plant parts were leaves and roots while the most common mode

of preparation was decoction. Plants with the highest UV were Musa paradisiaca for treating

fever and headache, Blumea balsamifera for cough, and Plectranthus scutellarioides also for

fever and headache. The highest ICF value was cited in the category 9, diseases of the visual

system with Euphorbia hirta (90% FL) as frequently used species for treating sore eyes.

SYNTHESIS

Sambong (Blumea balsamifera L.) is an aromatic shrub that has long been used in

traditional Philippine medicine to treat fever, cough, headache, boils, abdominal pain, and
 43

gaseous distention. This is a very common Philippine herbal flowering plant that is used as

medicine to treat wounds and cuts, rheumatism, anti-diarrhea, anti-spam, colds, and coughs. It's

also used to treat infected wounds, respiratory infections, and stomach aches.

This study will prove that Sambong leaves as an antibacterial soap acquire antibacterial and

antifungal activities against various infections and toxin-producing microorganisms, especially

Staphylococcus Aureus. These findings implied that Sambong leaves as an antibacterial soap

promoted wound healing and could feasibly be developed for use in the treatment and/or

prevention of microbial diseases.

Chapter III

RESEARCH METHODOLOGY
 44

In this chapter, researchers analyze the basic principles used to show the effectivity of

blumea balsamifera (sambong) leaves extract as an alternative antibacterial soap against

Staphylococcus Aureus.

RESEARCH DESIGN

Experimental research is an objective, systematic, and supervised investigation designed to

test the hypothesis and determine if the expected or desired phenomena can be achieved. The

experimental research method was used in the analysis because it was the best method for

establishing the cause-and-effect relationship within the given hypothesis. The study aimed to

evaluate and observe the efficacy of “blumea balsamifera” (Sambong) Leaves as an Alternative

Antibacterial Soap.

SOURCES OF DATA

A primary data source, according to Salkind, N.J. (2010), is an original data source, that

is, one in which the data are collected firsthand by the researcher for a specific research purpose

or project. The primary source in this study is the data gathered by the researchers through the

process of experimentation, data analysis utilizing statistical tools, observation, and

documentation. Cited articles, e-books, published journals, and related studies from the internet.

Some worked papers by researchers that contain correlated studies based on Staphylococcus

Aureus— disease, and also blumea balsamifera reviews were considered as the secondary source

of data, which was defined by Prachi, J. (2015) as the data that have been already collected by

and readily available from other sources.

RESEARCH LOCALE
 45

The study was conducted in the Philippines. The purpose of the experimental study is to

prove that blumea balsamifera (Sambong) leaves can be an alternative antibacterial soap to cure

wounds and cuts especially the Staphylococcus Aureus disease. The experiments were made and

observed at the researcher's house. The result of the product was evaluated and tested by the

researchers. All the information gathered was written in this study, and discussed through online

networks.
 46

Sambong Leaf
Plant Identification Description Authentication

Sambong Collection and Extraction

Cutting Boiling Extracting

Isolation of Bacteria
Description Identification Verification

Waste Handling
Proper Disposal Precautions Safety Measure

Interpretation of Result
Statistical Analysis and Evaluation

Figure 2: Research Process

 47

Preparation of Antimicrobial Leaf Extract

Preparation of Antimicrobial Leaf Extract the phases in creating Blumea Camphor leaves

extract as an antibacterial soap are the following: First, the researcher will collect a few leaves of

Sambong, which can be found anywhere, particularly in the backyard of the researcher's

grandparents. After collecting some leaves, put five cups of water in a pot, together with ten

medium-sized leaves, let it simmer until the water becomes color green like the leaves. When the

sambong leaf extract is cold and ready, drain it and pour it out in a container. In the last

preparation, test the sambong extract soap in various concentrations; 100%, 75%, %50%, and

25%. After the experiment, the researcher will be able to analyze the effectiveness of the

sambong extract at different levels. Consequently, the product will differentiate its content from

the positive control commercial products of sambong soap in markets. The result of this

experiment will be determined by using the weighted mean, frequency test, and also the t-test.

Preparation of Positive Control

Soaps and handwashes labeled as being “antibacterial” can be purchased from any

supermarket, but these commercial products rarely mention the antimicrobial spectrum of

activity. In the present study, the researchers evaluated six commonly used “antibacterial”

handwash solutions and five commonly used antibacterial toilet soaps to evaluate the spectrum

of their antimicrobial activity, without assessing the in vivo efficacy.

The rapid antimicrobial efficacy of the tested soaps was determined after 30 seconds

contact time against four strains: S. aureus (29213), S. epidermidis (SE 12), P. aeruginosa (Ps

AT) and C. albicans (C4), according to Shintre et al, 2006. The experiments were carried out in

triplicate and the means of Log10 reduction (reduction factor) were calculated. Ethyl alcohol
 48

solution (70%) was taken as the positive control. During the study, blind experiments were

carried out where the investigator was not being aware of the label or the active ingredient of the

product being tested.

Preparation of materials needed

Endemic throughout the tropics and subtropics of Asia, Blumea balsamifera (also known

as ngai camphor) is used widely as traditional medicine. Leaf extracts are directly applied to treat

eczema, dermatitis, skin injury, bruises, beriberi, lumbago, menorrhagia, rheumatism, and skin

injury. The materials needed to make an anti-bacterial soap are as follows: Sambong leaves are

the most needed because this is where the anti-bacterial soap for Staphylococcus Aureus will be

made of. The Extracts from Blumea balsamifera demonstrate a variety of bioactivities; including

antimalarial, antitumor, antifungal, and anti-obesity properties. Followed by 100 grams of soap

of glycerin so it won’t dry out the skin the way harsh treatments do to create a vicious cycle of

oil production, clogged pores, dry skin, and oil production. Also, 5 glasses of cooking oil and 5

glasses of water will also be a need in this experiment. On the other, the utensils that will be used

are plastic gloves, plastic pail, wooden ladle, glass or cup, strainer, knife chopping board,

cooking pot (preferably made of clay, enamel, stainless or glass), and importantly, plastic

molders that tend to be melted, cool and solidify, and then be melted again the soap without

burning.

Method and Procedure

 49

The natural and garden-fresh sambong leaves will be collected in Darasa, Tanauan City.

This medicinal plant was approved by the Department of Health (DOH) as a natural product that

can be used as a source for potential drugs. The DOH through the Philippine Institute of

Traditional Alternative Health Care (PITAHC) under Republic Act No. 8423 endorsed the use of

traditional medicines in the country. (Principe, Jose 2002) This method was modified from the

School of Science and Technology, Centro Escolar University, Manila, Philippines (2019). The

herbal plant called sambong will be boiled and extracted containing 740 mL of water. The cubes

of glycerin soap weighed 100 grams and boiled for 5 minutes. The second is to mix up the

extracted leaf. Afterward, gather all the ingredients and mix them all in the bowl. Then, pour it

into a pot and mix well by stirring continuously, using a wooden Ladle. Use only one direction in

mixing the mixture. Stir until the glycerin soap is dissolved, then pour five glasses of cooking oil

into the mixture. Continue stirring until the consistency of condensed milk is achieved. After

that, pour the soap mixture into desired plastic molders. Set aside and let it cool to harden. After

4-5 hours, remove the soap from the molder. and it is the finished product.

Proper Disposal and Handling of Staphylococcus Aureus—bacteria

The containers used for sambong extract and positive control will be dried after the

experiment and considered as waste will be put in a recyclable can. Tools and equipment used in

taking samples to the Staphylococcus Aureus bacteria and the workplace will be sanitized.

Proper handling of the specimen will be followed by the use of safety equipment such as gloves

and other PPE (Personal Protection Equipment). Sambong extracts and positive controls are

carefully put in clean and isolated containers to avoid contacts.

 50

Safety precautions and procedure

Staphylococcus aureus is a common etiologic organism in soft infections and may be

found on the skin of nearly 20% of healthy people. Staph bacteria are one of the most common

causes of skin infections and are the common cause of pneumonia, surgical wound infections,

and bloodstream infections. Therefore, proper safety precautions must be observed at all times.

Operations and safe use of all materials and equipment must be done. The proper use of personal

protective equipment such as safety goggles, a mask, rubber gloves, an apron, long sleeves, and

shoes that cover the entire feet (nothing open-toed) must be worn. Hair should be tied back and

away from the face. Protective eye gear or safety goggles should be worn at all times to prevent

anything from getting into the eyes. A mask is suggested for the mixing of the water and lye.

Rubber gloves should be worn during the whole soaping process. Always wear an apron. The

shirts should be long sleeve. Shoes must be worn. Nothing that is open-toed or leaves any portion

of your feet exposed. The pets and children should be away from the soaping area. When it

comes to crafting soap, protective gear is mandatory. It is also just as important to have a safe

and clean work environment. Ensure that all of the ingredients are already in the work area.

Researchers must be prepared because it is the one key factor in successful crafting. Once it gets

started, it is vital to stay in the work area. Leaving certain ingredients such as lye out in the open

can lead to very serious and dangerous situations. During the soap-making process, it is very

important to researchers to not do in rush. Since soap making is a science, and it is a must to

ensure that everything is measured out exactly. Soap recipes are measured by weight units, not

volume units. In other words, if a recipe calls for 8 oz. of coconut oil, it will need to weigh out 8
 51

oz. of coconut oil on scales. Take time and move methodically. The best way to work is in an

organized fashion. Also, while making soap, it is important for the researchers to be able to

concentrate and work uninterrupted. An unauthorized experiment or procedure must not do.

Statistical tool

In SOP 1, the statistical tools of frequency distribution and mean will be used to analyze

the results. The properties of blumea balsamifera” (Sambong) Leaves will be presented in

frequency distribution to demonstrate its potential as antibacterial soap. Although the weighted

mean will be used to depict the average result of each concentration. A T-test will be used in

SOP 2 and 3 to compare two means from the same individuals, objects, or relative units. It is

used to determine the efficacy of each concentration at various levels and the significance of the

differences.

Interpretation of data

This study was designed to evaluate the efficacy of blumea balsamifera (Sambong) leaves

decoction as an alternative antibacterial soap. The result of the product might be effective within

24 hours. It will be interpreted depending on different time intervals on different phases of trials.

At the end of the experiment, the outcome will define if it is truly the alternative cure for

Staphylococcus Aureus disease.

 52

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Aklan,
 53

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 54

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