Hair Growth Potential of Rice (Oryza sativa) Bran Oil as a

Shampoo bar tested on Male Albino Mice

A Thesis
Presented to the Faculty of the
(name of your school),
(city address)

In Partial Fulfillment
of the Requirements in the Subject
Practical Research 2
(Senior High School Department)

(Group member/s)

APRIL 2020

ABSTRACT
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Title: THE HAIR GROWTH POTENTIAL OF RICE (ORYZA

SATIVA) BRAN OIL AS A SHAMPOO BAR TESTED

ON MALE ALBINO MICE

Researchers:     (group member/s)

Institution: (your school).

Duration of the Study: 2019-2020

An average adult has 100,000 to 150,000 hair strands but loses about a

hundred per day (Gardner, 2017). Hair loss has been a problem to most adults,

especially at the ages 35 and above. With topical treatments for this condition,

this study was conducted to determine the Hair Growth Potential of the test

material: the Rice (Oryza sativa) bran oil as a formulated Shampoo bar tested on

Male Albino Mice, and the Black Beauty Shampoo as the Positive Control. An

experimental quantitative method was used to collect the needed data, through

the 3-applications processes under a 7-days observation period on 3 subjects for

the formulated test material, and another 3 for the positive control. The

formulated shampoo bar has shown more potential of hair growth than of the

positive control. The ANOVA statistical technique was used to determine the

significant difference of the potential hair growth of the male albino mice.

TABLE OF CONTENTS
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Page
Title page -------------------------------------------------------------------------------------- I

Fly Leaf ----------------------------------------------------------------------------------------- II

Acknowledgement ----------------------------------------------------------------------------

III

Abstract ----------------------------------------------------------------------------------------- IV

Approval Sheet ------------------------------------------------------------------------------- V

Table of Contents ---------------------------------------------------------------------------- VI

List of Tables ---------------------------------------------------------------------------------- IX

List of Figures --------------------------------------------------------------------------------- X

List of Plates -----------------------------------------------------------------------------------

XI

CHAPTER 1: INTRODUCTION

Background of the Study -------------------------------------------------------- 1

Review of Related Literature --------------------------------------------------- 3

Theoretical Framework ----------------------------------------------------------- 15

Conceptual Framework ----------------------------------------------------------- 16

Statement of the Problem -------------------------------------------------------- 17

Null Hypothesis --------------------------------------------------------------------- 17

Significance of the Study --------------------------------------------------------- 17

Definition of Terms ----------------------------------------------------------------- 19

CHAPTER 2: METHODOLOGY

Research Design ------------------------------------------------------------------- 20

Subjects ------------------------------------------------------------------------------- 21
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Ethical Considerations ------------------------------------------------------------- 21

Research Instruments ------------------------------------------------------------- 22

Data Collection ---------------------------------------------------------------------- 24

1. Collection and Preparation of the Test material ---------------- 24

2. Extraction of Rice Bran oil ------------------------------------------ 24

2.1 Materials for Extraction ------------------------------------ 24

2.2 Procedure ---------------------------------------------------- 24

2.3 Physical and Chemical characteristic ----------------- 25

3. Shampoo bar Formulation ------------------------------------------- 25

3.1 Ingredients ---------------------------------------------------- 25

3.2 Procedure ----------------------------------------------------- 26

4. Proper care of laboratory animals --------------------------------- 26

4.1 Cage Design ------------------------------------------------- 26

4.2 Food and Water --------------------------------------------- 26

4.3 Handling -------------------------------------------------------- 27

4.4 Temperature -------------------------------------------------- 27

5. Application to the laboratory Animals ----------------------------- 27

5.1 Shaving and antisepsis ------------------------------------ 27

5.2 Application of the study ------------------------------------ 28

6. Monitoring the Hair Growth of the Mice -------------------------- 28

7. Biosafety Laboratory Guidelines ----------------------------------- 29

8. Statistical Treatment -------------------------------------------------- 30

Scope and Limitation ----------------------------------------------------------------------- 31

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CHAPTER III: PRESENTATION, ANALYSIS, AND INTERPRETATION OF DATA

Changes of Potential Hair Growth from Bran Oil Shampoo Bar ------ 32

CHAPTER IV: SUMMARY, CONCLUSION, AND RECOMMENDATIONS

Summary of Findings ------------------------------------------------------------- 34

Conclusion --------------------------------------------------------------------------- 36

Recommendations ----------------------------------------------------------------- 37

References ------------------------------------------------------------------------------------ 39

APPENDICES

APPENDIX A ----------------------------------------------------------------------- 43

APPENDIX B ----------------------------------------------------------------------- 45

APPENDIX C ------------------------------------------------------------------------ 46

APPENDIX D ------------------------------------------------------------------------ 47

APPENDIX E ----------------------------------------------------------------------- 48

LIST OF TABLES

TABLES PAGE

1 Table for the Characteristics of Rice Bran Oil 25

2 Table of Ingredients 25

3 Table for the Effects of the Positive Control and Formulation 33

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LIST OF FIGURES

FIGURES PAGE

1 Rice (Oryza sativa) 3

2 Conceptual Paradigm of the Study 16

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LIST OF PLATES

PLATES PAGE

1 Maceration Process 50

2 ROTAVAP Process for Extraction 51

3 Shaving of Test Subjects 52

4 Formulation of Shampoo Bar 53

5 Application Process 54
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CHAPTER I

INTRODUCTION

Background of the Study

Hair grows everywhere on the human body, except the soles of our feet

and palms of our hands. It is made up of a protein called Keratin. According to

Gardner (2017), an average adult head has about 100,000 to 150,000 hair

strands, but loses about 100 strands a day. Hair Loss has been a problem to

most adults ages 35 and above. Various hair loss treatments are available in the

market that can cost up to $15,000 (Smith, 2017). However, people would want a

cheaper yet effective alternative in treating hair loss. The use of natural organic

products have been trend lately because these are much cheaper and healthier.

This study can give benefit for this problem, for this gives a potential treatment

for hair loss and also promotes the growth of a healthier and stronger hair.

According to the Hair Society, approximately 35 million males and 21

million females suffer from hair loss; beginning at the age of 35 with a loss rate of

40% for men, growing up to 70% by the time they reach 80. By age 60, women

suffering with hair loss will be in the range of 80%. An article from the Genetic

Home Reference stated that hair loss affects an estimated 50 million men and 30
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million women in the United States. It starts as early as a person's teens and risk

increases with age. According to the Philippine Council for Health Research and

Development, 50% of Filipino men will experience some form of hair loss by the

time they reach the age of 40 due to male pattern baldness, the reason is often

genetic, but some can be caused by nutritional deficiencies and lack of care.

Rice (Oryza sativa) bran oil is a potential treatment for hair loss.

According to Ali Reynolds (2017), the rice bran oil can bring a lot of benefits to

the hair with the help of its chemical constituents, Oleic Acid, Linoleic Acid,

Palmitic Acid, Stearic Acid, Vitamin B-group, and Vitamin E, promote hair growth

benefits like prevention of hypersensitivity, prevents and fights off dandruff, and

etc. Rice bran oil also gives various benefits to the hair because it has properties

and nutrients, such as the notability of the hypoallergenic properties that most

vegetables do not possess; antioxidants that help in protecting the hair from UV

exposure. This contains a high content of vitamins and fatty acids to prevent hair

loss. (Roos, et al., 2012)

Hence, this study explores the hair growth potential of Rice (Oryza sativa)

brain oil as a shampoo bar against hair loss. This study will give benefit to the

community by promoting a cheaper and healthier alternative hair treatment than

harmful chemicals costly procedures, in a way of conserving the environment by

alternatively using bio-wastes rather than throwing it all away.

Review of Related Literature

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This chapter provides information and related studies regarding our plant

material and its uses.

Figure 1. Rice (Oryza Sativa)

Kingdom: Plantae

Subkingdom: Tracheobionta

Superdivision: Spermatophyta

Division: Magnoliophyta

Class: Liliopsida

Subclass: Commelinidae

Order: Cyperales

Family: Poaceae

Genus: Oryza

Species: Oryza Sativa

Symbol: Orsa

Group: Monocot

Plant Description
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Rice (Oryza sativa) is an edible starchy cereal grain and plant which is

produced on its natural habitats, tropical marshes, but it has now grown into a

wide variety of Subtropical and Tropical habitats. Roughly one-half of the world’s

population, mainly all of the East and Southeastern Asia, is wholly dependent on

rice as their staple food; 95% of the world’s rice crop is consumed by humans.

The cultivated rice plant (Oryza sativa) is an annual grass of the family

Gramineae. It grows about 1.2 metres (4 ft.) in height, its leaves are long and

flattened, and its panicle, or inflorescence is made up of spikelets bearing flowers

that produce the fruit or grain. In planting rice plants, the seeds are sown into

prepared beds, the seedlings become 25-50 days old and are transferred to a

rice field or paddy that has been enclosed by levees and is submerged under 5-

10 cm (2-4 inches) of water, and remains submerged in the water during growing

season. After harvesting, the rice kernel, known as paddy or rough rice is

enclosed by a fiber coating called the Rice hulls of husks. The milling process

removes both the hulls and bran layer of the kernel, a type of rice wherein the

hulls were only removed are called brown rice, in which contains 8% protein and

is a good source of thiamine, niacin, riboflavin, iron, and calcium. Rice that is

milled to remove the bran is called white rice and it is greatly diminished in

nutrients. (Augustyn, 2018)

Rice is a staple in the meals of homes in Asian countries and some

European and Western countries. Rice is known to be the world’s second most

important crop, next to corn, about 482 million metric tons of husked rice is
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annually produced worldwide. Asian countries hold the largest contribution to the

world’s rice production, the global consumption of milled rice is approximately

477.77 million metric tons annually; 146 million metric units of which are

consumed by China.

The palay production in the Philippines had its ups and downs through the

years. A recent survey from the Philippine Statistics Authority (PSA) has stated

that the 2018 Philippine rice production has decreased by 2.2% from the

previous year. The October to December 2018 rice production which is at 7.16

million metric tons is about 2.2% lower than the 2017 rice production which is at

7.16 million metric units, but this matter didn’t affect the country’s inflation rate.

According to the statistics of the City Government of (location), the

annual rice production in (location) is said to be 3,822 metric tons according to a

2017 survey. Rice is the second major crop produced by the city, next to

coconuts and before corn. The barangay is known to be the biggest contributor of

rice production in (location), located in the (location) part of the City.

Rice Bran

Rice bran is a component of raw rice which is obtained when the starchy

endosperm of rice grain is removed during the milling process. It is the cuticle

existing between the rice and hulls of the paddy and it consists of embryo and

endosperm of the seeds of Oryza sativa. It constitutes 8% of the weight of the

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whole full grain and contains most of the nutrients. Rice bran is highly nutritious

and delivers a powerhouse of health supporting nutrients, but it is commonly

used as a low-level animal feed or be either thrown away (Qureshi, et al., 2000).

It has the potential to be used as an ingredient for food, since it has a good

amount of nutrients. However, bran must be stabilized immediately upon

production because of lipase, an enzyme that rapidly hydrolyzes oil to free fatty

acid (FFA) and glycerol, which results in a drastic quality reduction of rice bran.

These shortcomings were now overcome through destroying the lipolytic activity

using advance stabilizing technology, the co-product obtained from this is called

“stabilized” rice bran which has good taste and results in a readily soluble longer

shelf life. The primary means for the stabilization of rice bran includes enzyme

deactivation through heat treatment like microwave heating. The nutritional

composition and values of rice bran has led to the discovery of its varied health

benefits. Rice bran is also used to enrich food, due to its high nutrient content.

(Boshale and Vijayalakshmi, 2015).

Rice bran is a rice processing by-product. It is the outer coating of rice

grains which accounts for tons of food waste per year. It is composed of

numerous nutrients and bioactive substances that are able to reduce the risk of

non-communicable chronic diseases. It is also used as animal feed; however,

newer studies have proved that it actually brings numerous health benefits.

There are many more uses of rice bran other than agriculture and structural

uses. Rice Bran (extracted from the hull of rice) is commonly used for cooking
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because it is a healthy alternative to fatty, chemically processed oils made in a

lab. However, it also has benefits for your health, hair and skin. (Ferreira, et al.,

2019)

Rice Bran Oil

According to Anthony Morrocco, a founder of Morrocco Method Int’l, 2016,

rice bran oil is popular among Asian countries like China, Japan, and most

particularly in India. It is popularly known as “Healthy Oil”. Some studies claim

that Rice bran can be used for treating diabetes, high blood pressure, and many

more. Some people also apply it directly to the skin for an allergy skin rash called

eczema (ectopic dermatitis). It has numerous chemical constituents such as

Oleic Acid, Linoleic Acid, and etc. Rice Bran Oil is a well kept secret in Asia

(Japan specifically). Many women in Japan have been known to apply Rice Bran

Oil to their skin and are therefore known as “Rice Bran Beauties.” In general

health, Rice Bran Oil can be used for treating diseases such as high cholesterol,

diabetes, and AIDS. This can also help in strengthening the immune system and

can be used to help prevent heart disease and colon cancer. In today’s society,

the use of this as a soap base is now very popular. Rice Bran Oil has been

known to have many benefits for hair. It helps protect your hair from

sun/wind/cold damages, normalizes the oiliness of your scalp. strengthens your

roots, and tackles dandruff.

Rice bran oil is rich in micronutrients such as oryzanols, tocopherols,

tocotrienols, phytosterols, and dietary fibres. Due to problems associated with

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lipase enzyme, the high nutritional profile of rice bran was not used, which

reduced the quality of rice bran and rendered it unfit for human consumption.

Upon lipase enzyme stabilization, highly nutritious value-added rice bran

products can be extracted. It helps to reduce plasma cholesterol, decreases

serum cholesterol, decreases cholesterol absorption, and decreases platelet

aggregation through to the presence of antioxidants. This is also used to treat

hyperlipidemia, diseases of menopause, and increases muscle mass. The rice

bran's most widely accepted commodity is its oil which has outstanding

properties compared to other vegetable oils. (Sharma,et al., 2015)

Rice bran is the outer rice kernel layer and is a rice milling by-product that

accounts for 8 percent of all rice grain. This contains different nutrients such as

protein, carbohydrates, fats, dietary fiber, vitamins and minerals. It has a unique

blend of a number of phytochemicals such as amblezanol, tocopherol,

tocotrienol, phytosterol, etc. These compounds also act as free radicals of

antioxidants. It also has some antinutrients such as lipase, phytate, inhibitors of

trypsin, etc. Only by deactivating these anti-nutritional factors, particularly lipase,

which develops rancidity in rice bran, the effective use of rice bran can be done.

There are different ways of stabilizing the rice bran and enhancing its efficiency

and potential position. It can play an important role in reducing serum cholesterol,

hypertension, alleviating postmenopausal syndrome, enhancing insulin

sensitivity, reducing skin-related issues, and cancer cell growth. Rice bran oil and

wax are rice bran products. in which rice bran oil is more important because it is
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the concentrated source of alcohol and other antioxidants. The various food

products produced by adding processed rice bran help to boost the product's

nutrient content, textured properties and also improved product shelf life. (Dr.

Pramod, et al.,2016)

Rice bran oil has a high smoke point (450 degrees Fahrenheit) and is

quite suitable for dishes involving high temperatures. The goodness of rice bran

oil comes from its components. It contains y-oryzanol, a potent antioxidant, and

other organic chemical compounds like tocopherols and tocotrienols, which have

properties of vitamin E. Most of the benefits of this oil come from these

compounds, which is what we will discuss now. Rice bran oil contains inositol, a

carbohydrate compound that prevents dandruff and reduces split ends. It also

promotes hair health. The oil contains omega-3 and omega-6 fatty acids (though

omega-3 is only in small amounts) helps prevent premature greying of hair.

Studies show that linoleic acid and oryzanol also boost hair growth and

strengthen your tresses. These are the ways rice bran oil can keep you healthier.

But there is something else we need to know- the many other nutrients in the oil

that do the job well.

The benefits of the rice bran oil to the hair are: it prevents hypersensitivity,

prevents dandruff, fights against dandruff, thickens hair, and delays premature

grays. Prevents hypersensitivity, perhaps, the most notable feature of Rice Bran

Oil is its hypoallergenic properties, something that many vegetable oils do not
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possess. This makes the oil a great remedy for irritations and hypersensitivity.

Prevents dandruff, split ends are a common sign that your hair needs some TLC

– stat. The only remedy for split ends is to chop them off, but it doesn’t mean you

can’t prevent it from happening again. All the hair needs is a hair care regimen

that enhances the protective barriers of the hair against external aggressors.

Rice Bran Oil contains rich amounts of antioxidants that help protect the hair from

UV exposure. This oil also has a high smoking point, making it a natural heat

protectant. Fights dandruff, through a high vitamin E content, Rice Bran Oil

stimulates blood flow to the scalp and proper healing, which affect scalp health

and dandruff control. This is because the vitamin E in the oil is an antioxidant

itself. Like Cotton Protein and Kukui Butter, Rice Bran Oil helps fight off free

radicals and bacteria that’s causing the irritation. The Rice Bran Oil helps

alleviate the itching and discomfort caused by dandruff. Thickens hair, Rice Bran

Oil is also rich in omega-6 fatty acids and oleic acid. Combine them with the

antioxidants found in this wonder oil, and you have a hair growth super potion.

Applying Rice Bran Oil regularly to thinning spots will help stimulate blood

circulation in those areas, reviving the hair follicles to create strong and healthy

hair. Delays premature grays, finding grey hairs can be frightening for some as

it’s usually associated with aging. Contrary to popular belief, grey hair doesn’t

depend solely on age. It’s when our body starts producing less melanin – the

pigment that gives the strands their natural color – that causes a sudden onset of

grey hairs. This phenomenon is due to oxidative stress, a condition resulting from

the imbalance between free radicals and antioxidants. Rice Bran Oil supplies the
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hair and scalp great amounts of antioxidants to prevent the hair from losing

pigment due to oxidative stress. (Ali Reynolds, 2017)

Chemical Constituents of Rice Bran Oil

Oleic Acids, also known as Omega 9, a monounsaturated fatty acid, is a

chemical component of rice that has a lot of benefits such as: it can help in

maintaining the suppleness, softness and radiance of the hair, can stimulate the

growth of longer, thicker and stronger hair, can eliminate dandruff and thereby

support hair growth, and can boost immunity and exhibit antioxidant properties.

Linoleic Acid, also known as Omega 6, is the polyunsaturated fatty acid

that is the important source of energy. They can lead various benefits for the

hair such as: moisturizing the hair and promoting its growth, an effective

emulsifier in the formulation of soaps and quick-drying oils, exhibits anti-

inflammatory properties, promotes moisture retention on the hair and makes

oil feel thinner in consistency when used in an oil blend.

Palmitic Acid is a saturated long-chain fatty acid with a 16-

carbon backbone, has an emollient properties that can soften hair without

leaving a greasy or sticky residue.

Stearic Acid, also called as Octadecanoic Acid, is one of the most

common long-chain fatty acids. It is found in combined form in natural animal

and vegetable fats. It has cleansing properties that purge dirt, sweat, and
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excess sebum from hair and skin, and an ideal emulsifying agent that binds

water and oil. It also helps the product to remain potent when stored for a long

period of time. It conditions and protects the hair from damage without

diminishing luster or making it feel heavy. It also has exceptional cleansing

properties.

Vitamin B-group is a collection of eight water soluble vitamins that are

essential for various metabolic processes. However, it can lead to many

benefits such as retaining the ability of hair’s texture, color, and strength.

Vitamin E is a vitamin that dissolves fat. It is required for the proper

function of many organs in the body. It also helps to prevent moisture loss of

skin and hair, deeply cleanses pores and balances the oil production

According to the study of Choi, J.S., et al. (2014), the potential hair growth-

promoting activity of rice bran supercritical CO2 extract (RB-SCE) and major

components of RB-SCE, linoleic acid, policosanol, γ-oryzanol, and γ-tocotrienol,

were evaluated with the histological morphology and mRNA expression levels of

cell growth factors using real-time reverse transcriptase-polymerase chain

reaction (PCR) in C57BL/6 mice. RB-SCE showed hair growth-promoting

potential to a similar extent as 3% minoxidil, showing that the hair follicles were

induced to be in the anagen stage. The numbers of the hair follicles were

significantly increased. In addition, mRNA expression levels of vascular

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endothelial growth factor (VEGF), insulin-like growth factor-1 (IGF-1), and

keratinocyte growth factor (KGF) were also significantly increased and that of

transforming growth factor-β (TGF-β) decreased in RB-SCE-treated groups.

Among the major components of RB-SCE, linoleic acid and γ-oryzanol induce the

formation of hair follicles according to examination of histological morphology and

mRNA expression levels of cell growth factors. In conclusion, our results

demonstrated that RB-SCE, particularly linoleic acid and γ-oryzanol, promote hair

growth and suggests that RB-SCE can be applied as hair loss treatment.

Hair growth

Hair grows everywhere on the human body, except the soles of our feet

and palms of our hands. Hair is made up of a protein called Keratin which is

produced in hair follicles in the outer layer of skin. As new hair follicles are

produced, the old ones are then pushed out through the surface of the skin, with

the rate of 6 inches annually. According to Gardner (2017), an average adult

head has about 100,000 to 150,000 hair strands and loses about 100 strands a

day. On average, the hair on your head is growing about six inches a year, or

around half an inch a month. For about two to six years, each hair grows and

then stops and enters the telogen phase, during which it is gradually shed. You

lose about 50 to 100 hairs a day on average. Often, the follicle’s growth cycle

begins and a new hair begins to grow in after shedding. The life cycle of hair

occurs in 3 stages: Anagen, active hair growth that lasts between two to six

years, Catagen, transitional hair growth that lasts from two to three weeks, and
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Telogen, is a resting phase that lasts about two to three months. By the end of

the resting phase, the hair is shed and is replaced by new ones. The Manila

Times has reported the findings of a recent study which reveals that Asian men,

particularly those from the Philippines, are seriously concerned about hair loss.

Of those questioned, 69% of Asian men with hair loss felt ‘sad’ when other

people noticed or commented on their thinning hair. A further 30% of Filipino men

wished that they had more hair, revealing a direct link between hair loss and

confidence.

Shampoo Bars

Shampoo Bars are a solid alternative to your regular liquid shampoo. They

take effect like your regular liquid shampoo, wash off dirt and keep your hair

smooth. But unlike bottled liquid shampoo, it helps in conserving the environment

because one shampoo bar is equivalent to 80 washes or 3 medium sized bottles

of liquid shampoo. Shampoo bars help in cleansing and conditioning your hair

because of the natural organic ingredients used in making it.  According to the

U.S. Census and Simmons National Consumer Survey (NHCS) data, it claims

that 298.83 million Americans uses shampoo in 2017, most of these people are

using some form of bottled product.

Theoretical Framework

The theories are designed to predict, clarify and understand the

phenomena, to answer inquiries, and to expand the existing knowledge of the

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limits of the essential assumptions about the study. According to the study of

Roos, et al (2012), rice bran oil has various benefits to the hair because it has

properties and nutrients such as hypoallergenic properties and antioxidants that

helps in protecting the hair from UV exposure, and it contains a high content of

vitamins and fatty acids to prevent hair loss due to oxidative stress.

Conceptual Framework

The conceptual framework of this study shows the hair growth effects of

using a shampoo bar made from rice oil. This figure shows both the dependent
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and independent variables of the study. The independent variables of this study

are the Rice (Oryza sativa) bran oil as the test material and the Black Beauty

Liquid Shampoo as the positive control, This variable may show similar results to

the test material.

INDEPENDENT DEPENDENT
VARIABLE VARIABLE

Rice Bran Oil Hair Growth

(Test Material) (mm)

Black Beauty
(Positive Control)

Figure 2. Conceptual Paradigm of the Study.

Statement of the Problem

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This study aims to determine the benefits of the Rice (Oryza sativa) bran

oil as a shampoo bar for hair growth and specifically seeks to solve the following

question:

1. What are the changes in the hair growth using the formulated Rice

(Oryza sativa) bran shampoo bar?

Null Hypothesis

1. There is no significant difference in the fur growth of male albino mice

when administered with Rice (Oryza sativa) bran oil as a Shampoo bar

used for Hair Growth.

Significance of the Study

The study on the hair growth effects of Rice (Oryza sativa) bran oil would

be beneficial to the following:

Department of Health (DOH) The research about Rice (Oryza sativa)

bran oil can help them promote alternatives in using milling by-products in

terms of health and cosmetics.

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Cosmetological Industry This study will serve as a basis for the newly

innovated products because of the benefits of the rice (Oryza sativa) bran

oil that bring to the people.

Healthcare Professionals This study can give basis and guidelines for

advising and giving alternatives to people especially those who have hair

growth problems.

Researchers This study can provide the other researchers an ideas and

references for them to conduct an immense and further research related

to this study.

Patients This study is beneficial to the patients because it can provide

information on using alternatives for hair growth problems.

Readers This study can give ideas to the readers on the benefits of rice

(Oryza sativa) bran oil to hair growth.

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Definition of Terms

Rice Bran – also known as miller's bran, refers to the hard outer layers of

cereal grain. It consists of the combination of aleurone and

pericarp. Along with the germ, it is an integral part of whole grains, and is

often produced as a by-product of milling in the production of refined grains.

Rice Bran Oil - it refers to the oil extracted from the hard outer brown

layer of rice called chaff (rice husk).Rice bran oil is to be used as the main

component for shampoo bar.

Shampoo Bar – A rice bran oil will be formulated in a solid bar form. A

way of maintaining a healthy hair in a way that is also conserving the

environment.
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CHAPTER II

METHODOLOGY

This study is a research on the benefits and factors of Rice (Oryza sativa)

bran oil has helped in improving hair growth. This study is an experimental type

of quantitative research proving the benefits of (Oryza sativa) bran oil for hair

growth.

Research Design

The experimental method of research is used in this study to gather data

and information that was needed for the process of this study. The design

gathered and produced data was about the benefits of Rice (Oryza sativa) bran

oil as a shampoo bar for hair growth. A quantitative approach in research was

mainly used. It was used to gain a noting of the data that was gathered for and

from the experiment. (Babbie, 2010)

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Subjects

Male Albino White Mice are the test subjects that were used in conducting

the experiment of the study. It is one of the most prevalent animal subjects used

in studies because they are much more easily to cope up with. (Melina, 2010)

Ethical Considerations

In conducting this study, ethical considerations were observed. Ethics can

be thought of as the study of good conduct and grounds for making judgment

about what is good conduct. (Birch et al.,)

The researchers performed an experiment proper with the help of

professionals and the research adviser. All of the results were noted by the

researchers.

The researchers assured that before conducting the experiment, the

approval from the Research Adviser, Research Head of (your school name),

and Laboratory Personnel were made for the use and utilization of the

Laboratory.

The researchers assured that the Biosafety Laboratory Practices were

observed during the time of the experimentation and application process.

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The researchers assuredly followed the proper handling of the animal’s

subjects that was used in the study in accordance with the policy of the

Institutional Animal Care and Use Committee (IACUC).

The Albino white mice were separated and kept in a cage with free access

to food and water, the rice husks were separated in a container to avoid

contamination.

The researchers assured that all data were treated with appropriate

confidentiality and anonymity.

A letter of permission from the research adviser, dean’s office and the

laboratory in-charge with their approval of the utilization of the (your school

name) laboratory as the location on which the research experiment conducted.

Research Instruments

In our experimental research, “Rice (Oryza Sativa) Bran Oil Shampoo Bar

for Hair Growth”, we have utilized many instruments in conducting this study.

These are the following:

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Albino white mice were the medium of our experiment. A specific part of

the mice were shaved and was applied with the formulated shampoo bar,

and was observed for its hair growth effects.

Mice Cages enabled the mice to be secured and guarded for the whole

duration of the experiment.

Rice Bran (Oryza sativa) can boost hair growth. It contains inositol, a

carbohydrate compound that prevents dandruff and reduces split ends. It

also promotes hair health.

Organic essential Oil improves cellular metabolism when applied

topically, which stimulates hair growth by keeping the follicles active.

Soap Base a soap formula, can be soapy or slippery because it reacts

with acidic molecules of your skin called fatty acids.

Weighing scale was used in weighing the materials in the experiment.

Soap mold was used as a mold for the shampoo bar to hold its shape. 

Rotary Evaporator was used to extract the Rice Bran Oil.

Data Collection
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1. Collection and Preparation of the Plant

The Rice (Oryza sativa) bran used was collected from the rice milling area

of (location of rice milling area near you). Only the rice bran was

collected, which was the co-product of the second milling process of rice.

2. Extraction of Rice Bran oil

The Rice (Oryza sativa) Bran Oil was extracted using the Conventional

Solvent Extraction Method with the use of ethanol as the solvent.

2.1 Materials for Extraction

The materials used for the extraction of the rice bran oil were

Rotary Evaporator, 2 Erlenmeyer flask (500 ml), 1 Water Bath, 1 Hot

Plate, 5 Filter Papers, 2 Stirring Rods, 360 ml of Ethanol, and 60 grams of

Rice bran. (Krishnan, V.C.A., 2015)

2.2 Procedure

The rice bran powder (20 g) was mixed with ethanol with a 1:6 ratio

in a conical flask of 500 ml. The flask was put into the water bath with a

controlled temperature of 40°C for 15 min and allowed it to cool. After the

extraction, the extracts were filtered through a filter paper and then the

solutions collected were concentrated with a rotary evaporator to acquire

the rice bran oil. (Krishnan, V.C.A., 2015)

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2.3 Physical and Chemical characteristic to distinguish Rice (Oryza

sativa) bran oil

PARAMETER SPECIFICATION

Rice (Oryza sativa) bran oil (RBO)

COLOR has a characteristic of dark color.
(Jin, et.al, 2015)

Rice (Oryza sativa) bran oil has a

ODOR characteristic of clear and almost
odorless. (Kosher, 2020)

Table 1. Table for the Characteristics of Rice bran oil

3. Shampoo bar Formulation

3.1 Ingredients

PHASE INGREDIENT INCI % GRAMS

USED
1 Lavender Oil Lavender Extract Fragrance Oil 5.00 3.125

1 Castor Oil Castor (Ricinus Communis) Oil 5.00 3.125

1 Rice Bran Oil Rice (Oryza sativa) Bran Oil 8.00 5.00

2 Citric Acid Citric Acid 4.00 2.50

2 Cetyl Alcohol Cetyl Alcohol 4.00 2.50

2 SCS Sodium Coco Sulfate 74.00 46.125

TOTAL 100.00 62.375

Table 2. Table of Ingredients (Wright, 2017)

3.2. Procedure

Weighed out the Lavender oil, Castor oil and Rice Bran oil and

mixed in a double boiler or a hot plate. Once done, take liquid off the heat.
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Weighed out and added the sodium coco sulphate and the rest of the

ingredients. Stirred well, mold the and left to set. (Wright, 2017)

4. Proper Care of Laboratory Animals

4.1 Cage Design

It is suggested that the number of mice per cage is maximum of 5.

Mice housed in groups should have a cage height of no less than 5 inches

(12.7 cm) and a floor area per mouse is 6 to 15 inches (38.7 to 96.7 cm)

squared, depending on body weight. (Norins, 2012)

4.2 Food & Water

There was a safe and ample supply of water for mice at all times.

Mice can contaminate their water if it is supplied in a bowl or trough, so it

is advised to supply mice with water in adequately sized bottle feeders

that can accommodate the number of mice in the cage. Mice are

omnivores, eating a wide variety of plant and animal products, whether

necessary or provided. Mice are smart animals, and having a variety of

different foods which will help enrich their environment and minimize

boredom or harm. Only in very small quantities should cereals, seeds,

grains, biscuits, sweet food, bread, pasta and rice be fed as treats.

(Fawcett, 2012)

4.3 Handling
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When handling mice, handlers did it in a quiet and calm manner,

treating mice with extreme care to prevent distress and injury to the animal

and handler. The mice were picked up by almost fully closing the hand

around them and held tightly but without exerting unnecessary pressure.

The base of the tail safely lift the adult mice but never the tip. Once raised,

always immediately place the mouse on a firm surface such as table or

back of the hand while still holding. (Fawcett, 2012)

4.4 Temperature

The housing temperature for laboratory mice in the research facility

were between 20–26 °C; while the thermoneutral ambient temperature for

mice is 30–31°C. (Kozloski, 2019)

5. Application to the laboratory Animals

5.1 Shaving and antisepsis

The mouse's body part was shaven to determine the effect of the

made shampoo bar. This was performed with the use of an electric

shaver, with the help of a veterinarian. All rasped hairs were cleaned away

to prevent contamination.

5.2 Application of the study

We tested the Rice (Oryza Sativa) bran oil as an effective shampoo

bar for hair growth by applying it to the shaven body part of the Albino

mice. There were total of 6 albino mice used; the first three mice were
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used to test the hair growth potential of the formulated shampoo bar with

Rice Bran Oil, and the next three mice were used for the positive control,

the Black Beauty which is a shampoo for hair growth that is available on

the market. There were a total of 3 applications processeds during the 7-

day observation process,

6. Monitoring the hair growth of the Mice

The skin of the animals, usually the dorsal part, was observed everyday

in order to notice the hair regrowth period and the hair regrowth pattern. The hair

regrowth potential scores that used were: 0 = no hair growth, 1 = less than 20%

of hair growth, 2=20–39% of hair growth, 3=40–59%of hair regrowth, 4=60–79%

of hair regrowth, 5=80–100% of hair regrowth. Researchers also used self-

designed scales of hair regrowth, such as: Type 4 (high hair density, full, thick

fur), Type 3 (moderate hair density with no visible skin area), Type 2 (low hair

density, with the visualization of the skin), Type 1 (uneven hair growth on the test

area, skin easily seen). The hair growth of the mice was observed for 7 days.

(Vegesna, et al., 2002)

7. Biosafety Laboratory Guidelines

Researchers are expected to follow Biosafety recommendations in the

Laboratory and animal handling. (Lab Manager, 2017)

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1. Safety Laboratory gears such as (masks, gloves, lab coats, & etc.)

should be worn at all times inside the laboratory.

2. Hands should be washed before and after all laboratory operations.

3. Wearing of laboratory gear and clothing outside of the laboratory is

forbidden.

4. Open footwear should not be worn inside the laboratory.

5. Eating, or storing food and drinks inside the Laboratory work areas

are forbidden.

6. Used protective laboratory clothing must not be stored inside

lockers or cupboards.

7. You should not lift any glassware, solutions, or any other types of

apparatus above eye level.

8. Never smell or taste any chemicals.

9. If any chemicals splashes into your eye or gets in contact with your

skin, immediately flush the area with running water for at least 20 minutes.

10. Do not let any solvent or chemicals come in contact with your skin.

11. Flammable chemicals should be only used in a fume hood.

12. Never mix chemicals in sink drains.

13. Clean up right away if any spill occurs.

14. Ensure that all chemical wastes are properly disposed.

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The Standard Practices on Animal Biosafety Level 1 (ABSL-1) should be

observed:

1. Researchers are advised of the possible hazards, and are required to read

and follow the instructions on practices and procedures.

2. Eating, drinking, smoking, and applying cosmetics are prohibited inside

the animal room procedure rooms or lab.

3. Extra precaution is required in using sharp tools.

4. Gloves must be worn to prevent direct skin contact with contaminated,

infectious, and hazardous materials, and when handling animals.

5. Researchers must wash their hands after handling animals, after removing

gloves, and before leaving the animal facility.

8. Statistical Treatment

The experiment used was the ANOVA statistical technique to determine

the significant difference in the potential hair growth of the male albino mice from

the test material, which is the formulated rice bran oil shampoo bar and the

positive control, the black beauty shampoo. Analysis of Variance (ANOVA) is a

statistical technique that is used to check if the means of two or more groups are

significantly different from one another, it checks the impact of one or more

factors by comparing the means of different samples. (Singh, 2018)

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Scope and Limitation of the Study

8
The subject of the study was the albino mice that were used for the topical

experiment using a formulated Shampoo Bar, with Rice (Oryza sativa) bran oil

that was produced from the experiment for 3 weeks.

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Chapter III

PRESENTATION, ANALYSIS, AND INTERPRETATION OF DATA

This chapter presents the data obtained during the experimentation phase

of the study. The results are presented in the succeeding tables with

corresponding explanations and discussions.

Changes from Bran Oil Shampoo Bar on Hair Growth

Table 3

Effect of Rice (Oryza sativa) bran oil as a Shampoo Bar

and Positive Control for Hair Growth

Treatment Trial Hair growth, mm

Number Day Day Day Day Day Day Day

1 2 3 4 5 6 7

1 0 1 2 2 3 3 3

Positive control 2 0 0 0 0 1 1 1

3 0 0 0 0 1 1 1

1 0 0 1 1 2 2 2

Formulation 2 0 1 2 2 2 2 2

3 0 1 2 2 2 2 3
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Table 3. Table for the Effects of the Positive Control and Formulation.

Legend:
SCORES:
0 = No Hair Growth
1 = Less than 20%
2 = 20-39%
3 = 40-59%
4 = 60-79%
5 = 80-100%

The formulated shampoo bar was applied in two different groups (positive

control and treatment group) of mice (test animal). Each group is composed of

three mice which were monitored up to 7-day period. Results of the test showed

the hair growth potential of positive control and formulated shampoo bar from

bran oil showing a maximum hair length of 1.5mm for both treatments with an

equivalent score of 3.0. It means that the results of the positive control and

formulation diligently appeared during the third day of the observation, showing a

maximum growth of 1.5 mm and an equivalent score of 3, with a 40-59% of

visible growth for both treatments. According to a similar study of Umar (2019),

the hair growth potential of the rice bran extract in his study which was compared

to be similar to 3% minoxidil, based on all their findings, the researchers

concluded that the rice bran extract (RB-SCE) is a variant agent in treating hair

loss. The result of our study showed similar results as the study conducted by

Choi, J.S. (2014), that the potential hair growth-promoting activity of rice bran

supercritical CO2 extract and the rice bran extract (RB-SCE) from the study of

Umar (2019), has shown similar results to the formulated Rice (Oryza sativa)
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bran oil shampoo bar yielding a 40-59% of visible hair growth during the 7-day

observation period

CHAPTER IV

SUMMARY, CONCLUSION AND RECOMMENDATIONS

The goal of this chapter is to sum up the analysis undertaken. The

description includes an overview of the study's question, methodology, and

findings. The conclusion supports the recommendation which lays to the

foundation of the further research to be conducted in relation to the study.

Summary of Findings

This study was conducted to evaluate the hair growth potential of rice bran

oil as shampoo bar on albino male mice.

The extraction of the Rice (Oryza sativa) bran oil occurred on the (date)

of the year 2020, with the use of the Conventional Solvent extraction method

using 95% Ethanol as the solvent. The rice bran oil was mixed with ethanol with

a 1:6 ratio and was put in a water bath with a controlled temperature of 40°C for

15 minutes. The mixture was then filtered through a filtered paper, the collected

solution was then concentrated with a Rotary Evaporator to acquire the rice bran
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oil. A separatory funnel was then used to separate the rice bran extract from the

rice bran oil.

As for the formulation of the Rice (Oryza sativa) bran oil shampoo bar,

the following ingredients have been used: Lavender Oil, Castor Oil, Rice Bran

Oil, Citric Acid, Cetyl Alcohol, and Sodium Coco Sulfate. The process started

out by mixing the Lavender, Castor and Rice bran oil on top of a hot plate, we

then added the sodium coco sulfate along with the cetyl alcohol and citric acid to

the oil mixture. We then left it out in a mold to let it set.

The potential hair growth-promoting activity of rice bran oil is evaluated

by comparing the levels of cell growth factors from the skin of white male albino

mice. The results were evaluated statistically using the One-Way Analysis of

Variance (ANOVA). It has shown that during the application between the

positive control (black beauty shampoo) and the formulation of the Rice (Oryza

sativa) bran oil as shampoo bar), the visible hair growth has appeared as of day

3 garnering a percentage of at most 20% of hair growth. Among the mice that

were evaluated for the positive control, mice one has more positive effects on

hair growth, compared to mice two and three. While the mice for the test

material has shown that mice three has more positive effects on hair growth.

According to the total results from day one to day seven of observations, the

subjects for the test material have more noticeable effects on the hair growth

than the positive control.

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The statistical results have shown the difference of the hair growth of

male albino mice applied with the positive control (black beauty shampoo) has a

mean of 0.9524 and a SD of 1.07127, and the test material which was the rice

(Oryza sativa) as a shampoo bar has shown the results with the mean of 1.4762

and an SD of 0.87287.

Conclusion

This study aims to evaluate the hair growth potential Rice (Oryza sativa)

bran oil as shampoo bar on male albino mice. This study has also shown the

benefits and eligibility of Rice (Oryza sativa) bran oil as a safe cosmetic

ingredient. Based on the findings, the following conclusion is drawn by the

researchers of the study:

1. The changes that have occurred using Rice (Oryza sativa) bran oil as a

shampoo bar for hair growth that was yielded during the 7-day treatment

and observation were visible. The results of the formulated shampoo bar

appeared as early as Day 3 with at most 20% of hair growth, which later

on showed 20-59% of visible hair growth as of the last day of observation.

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Recommendations

After obtaining the results discussed above, the answer to the problems

presented in the Hair Growth Potential activity of the Rice (Oryza sativa) Bran Oil

as Shampoo Bar tested on Male Albino Mice, the researchers had the following

recommendations to further improve the study:

1. To the future Researchers, we would gladly recommend this study as

your basis. We suggest that you may use the same plant but with a

different variety. Through this you may be able to compare the rice bran

oil's hair growth potential ability with the use of other varieties and

methods.

2. To the Consumers, we suggest the effectiveness of the rice (Oyza sativa)

bran oil as a potential hair growth treatment. It is a possible and a

cheaper alternative for hair growth treatment other than costly hair

treatments and procedures currently available in the market.

3. To companies and agencies related to trichology, we recommend this

study as a basis to take the lead in enhancing and encouraging the use of

a natural products such as Rice (Oryza sativa) bran oil as a local and

natural remedy for the treatment of hair loss, to promote hair growth, and
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to also inform consumers about the safety, affordability and effectiveness

of rice bran oil.

4. To the pharmaceutical industry, we suggest that the result of this study

will serve as a basis on finding effective hair growth treatments with the

use of natural organic products, in a way of also helping consumers

conserve the environment.

5. To the Department of Health (DOH), we would highly recommend our

study as a basis in using rice bran oil as an effective hair growth

treatment that can help and encourage citizens suffering hair loss through

using cheaper yet effective alternatives of hair growth potential products

that also conserve the environment, and be widely available locally and

exported internationally
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reveals-psychosocial-side-effects-of-hair-loss-in-filipino-men/

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Extract Prepared by Supercritical Carbon Dioxide Fluid. Retrieved from

https://www.ncbi.nlm.nih.gov/pubmed/24389480

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Location and Extent. Retrieved from https://tagumcity.gov.ph/agricultural-a

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Food, and Industrial Uses, and Bioactivities in Humans, Animals, and

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Cells. Retrieved from https://datapdf.com/rice-brans-rice-bran-oils-and-rice-hulls-

composition-foodan.html?fbclid=IwAR3TqWL78CaX6ngQE50FIUU0qeoal

hHiJd_J%iY9BiZHD4IOI6g7xl_fUQg

Ferreira, S.C., et al. (2019) Encyclopedia of Food Security and Sustainability.

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81005965221409

Garba, U., et al. (2017) Extraction and Utilization of Rice Bran Oil: A

Review. Retrieved from https://www.researchgate.net/publication/3193540

31_Extraction_and_utilization_of_rice_bran_oil_A_review_extraction_and

_utilization_of_rice_bran_oil_A_review

Gardner, S.S. (2017) Understanding Hair Loss - The Basics. Retrieved from

https://www.webmd.com/skin-problems-and-treatments/hairloss/understan

ding-hair-loss-basics#1

Gotter, A. (2019) Essential Oils for Hair. Retrieved from https://www.healthline.co

m/health/essential-oils-for-hair-growth

Jin, J., et al. (2015) Production of Rice Bran Oil with Light Color and High

Oryzanol Content by Multi-stage Molecular Distillation. Retrieved from

https://www.researchgate.net/publication/284244635_Production_of_Rice

_Bran_Oil_with_Light_Color_and_High_Oryzanol_Content_by_Multistage

_Molecular_Distillation

Kosher (2020) Rice Bran Oil. Retrieved from https://www.aot.de/en/productde

tails/reiskleieoel.html
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Kozloski, G. (2019) Mice and Rats: Housing and Temperature Handling.

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Temperature-and-Handling.html

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d_Extraction_of_Oil_from_Rice_Bran

Martinez, B. (2013) Rice Bran Oil. Retrieved from https://www.brennta

g.com/media/documents/bsi/product_data_sheets/life_science/textron_nat

ural_oils/rice_bran_oil_tx008272_pds.pdf

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Bar for Cage Sizes. Retrieved from http://resadmin.uah.edu/resadminweb

/documents/uah_only/New_OLAW_Guide_Sets_for_Cage_Sizes.pdf

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January 2019 Round. Retrieved from https://psa.gov.ph/ricosit-main

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atista.com/topics/1443/rice/

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d0fa011dec7e1041f81.pdf?fbclid=IwAR0TBSYA1zAJPSzPagLMbv7hCIEo

1GeNMcPIWvynqQkV3C94LYuVCBfT11Y

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-loss-17/hair-transplants

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Umar, S. (2018) Is Rice Bran for Hair Beauty and Growth Help or Hype?

Retrieved from https://ugro.com/is-rice-bran-for-hair-beauty-and-growth-

help-or-hype/?fbclid=IwAR3UcFNjAmsdRyqFVAoSYqpToDfXfshvJjerBI8lr

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Unwrapped Life (2019) Top Eight Reasons to make the switch to Solid

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h-to-solid-shampoo-bars-and-conditioner-bars-too
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Vegesna, V., et al. (2002) Vitamin D 3 Analogs Stimulate Hair Growth in Nude

Mice. Retrieved from https://www.researchgate.net/publication/11064964

_Vitamin_D_3_Analogs_Stimulate_Hair_Growth_in_Nude_Mice

Wright, R. (2017) Shampoo Bars Tutorial, Retrieved from https://www.botanical

formulations.com/blog/shampoo-bars-tutorial

APPENDIX A
Laboratory Request

(Date)

(Name of your school’s custodian)

General Laboratory Custodian
(Name of your school) 
(city address)

Dear Ma’am, 

Greetings!

We, the researchers from Grade 12 - STEM of (your school name) would like to
ask for your permission from your good advice to allow us to use the laboratory
for the conduct our experimentation intended for our research entitled “Hair
Growth Potential of Rice (Oryza sativa) as a Shampoo Bar tested on Male
Albino Mice” as a part of our requirements for capstone.

Thank you and God Bless.

Respectfully Yours,
(Group member/s)
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Noted by:

(name of your research adviser)

   Capstone teacher

(Name of your QS)

 Qualified Scientist

(Name of your research director)

     Research Director

(chemicals and apparatus na hiniram niyo)

Wednesday:
● Soxhlet Apparatus
● Beaker 100 ml-4pcs
● Petroleum Ether – 400ml
● Stirring rod - 2pcs 

Friday:
● Rotavap 
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APPENDIX B
Plant Authentication Letter

(date)

Department of Agriculture  
(address)

Dear Ma’am/Sir,

Greetings!

This letter is to request your permission and assistance to verify/authenticate our

material which is a Rice Bran (Oryza sativa) of the variety 7-tonner for the
conduct of our undergraduate thesis entitled “Hair Growth Potential of Rice
(Oryza sativa) as a Shampoo bar tested on Male Albino Mice” 

We are hoping for your kind consideration and positive response, thank you and
God Bless!

Respectfully yours,

(Group member/s)

Noted by:
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  (name)     (name )
   Qualified Scientist                      Research Teacher

 (name)          name)  

   Research Director                  Senior High School Principal

APPENDIX C
Letter of Request for Veterinary Certification

February 17, 2020

(Name)
City Veterinarian
(Address)
Province of (name of your province)

Greetings with love and bliss!

We the researchers of Science ,Technology, Engineering and Mathematics (STEM)

of (your school name) to Check up mice for our research entitled “Hair Growth
Potential of Rice (Oryza sativa) bran oil as a Shampoo bar tested on Male Albino
Mice ”.

Hoping for your kind consideration.

Sincerely yours,

(group member/s)

Noted By:
(name )
Research Adviser
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Name) .
Senior High Principal

APPENDIX D

LETTER OF REQUEST FOR ANIMAL HOUSE

(Name)
Research Director
(School)
(address)

Dear Ma'am,

Greetings!

We the researchers from grade XII – Science, Technology, Engineering, and

Mathematics (STEM) of(school name), would like to ask for your permission to
use the animal house in (place) to maintain the well-being of our Male albino
mice in our study, “Hair Growth Potential of Rice (Oryza sativa) bran oil as a
Shampoo bar tested on Male Albino Mice".

Thank you and more power!

Respectfully yours,

(Group member/s)

Noted By:

(Name of your QS)

Qualified Scientist
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(Name)
Senior High School Principal

Appendix E
Statistics Output

A. Description of the Extracted Rice Bran Oil

PARAMETER SPECIFICATION

COLOR Dark in Color, hazy and mahogany colored

ODOR Has no odor; odorless

Table 4. Table for the Description of the Extracted Rice bran oil

B. Data of the Observation Process

Legend:

SCORES: TYPE:
0 = No Hair Growth 4 = High hair density, full and thick fur
1 = Less than 20% 3 = Moderate hair with no visible skin area
2 = 20-39% 2 = Low hair density, with visualization of the skin
3 = 40-59% 1 = Uneven hair growth, skin is easily seen
4 = 60-79% 0 = No hair growth
5 = 80-100%
MICE: 1 (Positive Control)

DAY SCORE TYPE REMARKS

1 0 0 No hair growth

2 1 1 Slightly visible hair growth

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3 1 2 Hair grew by 0.5 mm

4 2 2 Hair grew by 1 mm

5 3 3 Hair grew by 1.5 mm

6 3 3 No changes since the previous observation

7 3 3 No changes since the previous observation

Table 5. Table of Observation for Mice 1.

MICE: 2 (Positive Control)

DAY SCORE TYPE REMARKS

1 0 0 No hair growth

2 0 0 No hair growth

3 0 0 No hair growth

4 0 0 No hair growth

5 1 1 Hair grew by 0.5 mm; uneven hair growth

6 1 1 No changes since the previous observation

7 1 1 No changes since the previous observation

Table 6. Table of Observation for Mice 2.
MICE: 3 (Positive Control)

DAY SCORE TYPE REMARKS

1 0 0 No hair growth

2 0 0 No hair growth

3 0 0 No hair growth

4 1 1 Slight uneven hair growth

5 1 1 No changes since the previous observation

6 1 1 No changes since the previous observation

7 2 1 Hair grew by 1 mm; uneven hair growth

Table 7. Table of Observation for Mice 3.
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MICE: 4 (Test Material)

DAY SCORE TYPE REMARKS

1 0 0 No hair growth

2 0 0 No hair growth

3 1 1 Slight uneven hair growth

4 2 1 Slight uneven hair growth with visible skin

5 2 2 Hair grew by 1 mm; uneven hair growth

6 2 2 No changes since the previous observation

7 2 2 No changes since the previous observation

Table 8. Table of Observation for Mice 4.
MICE: 5 (Test Material)

DAY SCORE TYPE REMARKS

1 0 0 No hair growth

2 0 0 No hair growth

3 1 1 Slight uneven hair growth with visible skin

4 1 2 Hair grew by 0.5 mm; uneven growth

5 2 2 Hair grew by 1 mm; uneven hair growth

6 2 2 No changes since the previous observation

7 2 2 No changes since the previous observation

Table 9. Table of Observation for Mice 5.
MICE: 6 (Test Material)

DAY SCORE TYPE REMARKS

1 0 0 No hair growth

2 1 1 Slight uneven hair growth with visible skin

3 1 2 Slight uneven hair growth; less visible skin

4 2 2 Hair grew by 1 mm; uneven with visible skin

5 3 2 Hair grew by 1.5 mm; uneven hair growth

6 3 2 No changes since the previous observation

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7 3 3 Even hair growth with no visible skin

Table 10. Table of Observation for Mice 6.

Table 11

ANOVA on Hair Changes Caused by Positive Control and Formulated Bran

Oil Shampoo Bar in Various Monitoring Days

Types P Remark

of Std. F val s*

Treatme Mea Deviati val ue

nt Monitoring Time n on ue

Types of Positive Control .952 4.8 0.0 Significa

Treatmen 1.07127
t 4 4 36 nt

Formulated topical 1.47

.87287
Solution 62

Day 1 .000
.00000
0

Day 2 .500
.54772
Monitori 0

ng Time Day 3 1.16 .98319 5.6 0.0 Significa

67 7 01 nt
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Day 4 1.16
.98319
67

Day 5 1.83
.75277
33

Day 6 1.83
.75277
33

Day 7 2.00
.89443
00

*Calculation was performed at the 0.05 level of significance

Table 11. Table for the Changes caused by the Positive Control and Formulation.

Table 12

Post Hoc Multiple Mean Comparison Test of Hair Changes

in Various Monitoring Days

Mean P Remarks*

(I) Monitoring (J) Monitoring Difference val

Time Time (I-J) ue

Day 1 Day 2 .27 Not

-.5000
1 significant

Day 3 .01 Significant

-1.1667*
4

Day 4 -1.1667* .01 Significant

4
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Day 5 .00 Significant

-1.8333*
0

Day 6 .00 Significant

-1.8333 *

Day 7 .00 Significant

*
-2.0000
0

Day 2 Day 1 .27 Not

.5000
1 significant

Day 3 .14 Not

-.6667
6 significant

Day 4 .14 Not

-.6667
6 significant

Day 5 .00 Significant

-1.3333*
6

Day 6 .00 Significant

-1.3333*
6

Day 7 .00 Significant

-1.5000*
2

Day 3 Day 1 .01 Significant

1.1667*
4

Day 2 .14 Not

.6667
6 significant

Day 4 .0000 1.0 Not

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00 significant

Day 5 .14 Not

-.6667
6 significant

Day 6 .14 Not

-.6667
6 significant

Day 7 .07 Significant

-.8333
2

Day 4 Day 1 .01 Significant

1.1667*
4

Day 2 .14 Not

.6667
6 significant

Day 3 1.0 Not

.0000
00 significant

Day 5 .14 Not

-.6667
6 significant

Day 6 .14 Not

-.6667
6 significant

Day 7 .07 Not

-.8333
2 significant

Day 5 Day 1 .00 Significant

*
1.8333
0

Day 2 .00 Significant

1.3333 *

6
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Day 3 .14 Not
.6667
6 significant

Day 4 .14 Not

.6667
6 significant

Day 6 1.0 Not

.0000
00 significant

Day 7 .71 Not

-.1667
1 significant

Day 6 Day 1 .00 Significant

1.8333*
0

Day 2 .00 Significant

1.3333*
6

Day 3 .14 Not

.6667
6 significant

Day 4 .14 Not

.6667
6 significant

Day 5 1.0 Not

.0000
00 significant

Day 7 .71 Not

-.1667
1 significant

Day 7 Day 1 .00 Significant

2.0000*
0

Day 2 1.5000* .00 Significant

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Day 3 .07 Not

.8333
2 significant

Day 4 .07 Not

.8333
2 significant

Day 5 .71 Not

.1667
1 significant

Day 6 .71 Not

.1667
1 significant

*Calculation was performed at the 0.05 level of significance

Table 12. Table for the Post Hoc Multiple Comparison.

Descriptive Statistics

Dependent Variable: Hair Changes

Types of Treatment Monitoring Time Mean Std. Deviation N

Positive Control Day 1 .0000 .00000 3

Day 2 .3333 .57735 3

Day 3 .6667 1.15470 3

Day 4 .6667 1.15470 3

Day 5 1.6667 1.15470 3

Day 6 1.6667 1.15470 3

Day 7 1.6667 1.15470 3

Total .9524 1.07127 21

Formulation Day 1 .0000 .00000 3

Day 2 .6667 .57735 3

Day 3 1.6667 .57735 3

Day 4 1.6667 .57735 3

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Day 5 2.0000 .00000 3

Day 6 2.0000 .00000 3

Day 7 2.3333 .57735 3

Total 1.4762 .87287 21

Total Day 1 .0000 .00000 6

Day 2 .5000 .54772 6

Day 3 1.1667 .98319 6

Day 4 1.1667 .98319 6

Day 5 1.8333 .75277 6

Day 6 1.8333 .75277 6

Day 7 2.0000 .89443 6

Total 1.2143 1.00087 42

Table 13. Table for Descriptive Statistics

Tests of Between-Subjects Effects

Dependent Variable: Hair Changes

Type III Sum

Source of Squares df Mean Square F Sig.

Corrected Model 24.405a 13 1.877 3.154 .005

Intercept 61.929 1 61.929 104.040 .000

TT 2.881 1 2.881 4.840 .036

MT 20.238 6 3.373 5.667 .001

TT * MT 1.286 6 .214 .360 .898

Error 16.667 28 .595

Total 103.000 42

Corrected Total 41.071 41

a. R Squared = .594 (Adjusted R Squared = .406)

Table 14. Table for the Tests of Between-Subjects Effects

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Estimated Marginal Means

1. Types of Treatment

Dependent Variable: Hair Changes

Std. 95% Confidence Interval

Types of Treatment Mean Error Lower Bound Upper Bound

Positive Control .952 .168 .608 1.297

Formulation 1.476 .168 1.131 1.821

Table 15. Table for Types of Treatments

2. Monitoring Time

Dependent Variable: Hair Changes

Std. 95% Confidence Interval

Monitoring Time Mean Error Lower Bound Upper Bound

Day 1 .000 .315 -.645 .645

Day 2 .500 .315 -.145 1.145

Day 3 1.167 .315 .521 1.812

Day 4 1.167 .315 .521 1.812

Day 5 1.833 .315 1.188 2.479

Day 6 1.833 .315 1.188 2.479

Day 7 2.000 .315 1.355 2.645

Table 16. Table for the Monitoring Time

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Post Hoc Tests

Mean P Remarks*

(I) Monitoring (J) Monitoring Difference val

Time Time (I-J) ue

Day 1 Day 2 .27 Not

-.5000
1 significant

Day 3 .01 Significant

-1.1667*
4

Day 4 .01 Significant

-1.1667*
4

Day 5 .00 Significant

*
-1.8333
0

Day 6 .00 Significant

-1.8333 *

Day 7 .00 Significant

-2.0000*
0

Day 2 Day 1 .27 Not

.5000
1 significant

Day 3 .14 Not

-.6667
6 significant

Day 4 -.6667 .14 Not

6 significant
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Day 5 .00 Significant
-1.3333*
6

Day 6 .00 Significant

-1.3333*
6

Day 7 .00 Significant

-1.5000*
2

Day 3 Day 1 .01 Significant

1.1667*
4

Day 2 .14 Not

.6667
6 significant

Day 4 1.0 Not

.0000
00 significant

Day 5 .14 Not

-.6667
6 significant

Day 6 .14 Not

-.6667
6 significant

Day 7 .07 Significant

-.8333
2

Day 4 Day 1 .01 Significant

1.1667*
4

Day 2 .14 Not

.6667
6 significant

Day 3 .0000 1.0 Not

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00 significant

Day 5 .14 Not

-.6667
6 significant

Day 6 .14 Not

-.6667
6 significant

Day 7 .07 Not

-.8333
2 significant

Day 5 Day 1 .00 Significant

1.8333*
0

Day 2 .00 Significant

1.3333*
6

Day 3 .14 Not

.6667
6 significant

Day 4 .14 Not

.6667
6 significant

Day 6 1.0 Not

.0000
00 significant

Day 7 .71 Not

-.1667
1 significant

Day 6 Day 1 .00 Significant

*
1.8333
0

Day 2 .00 Significant

1.3333 *

6
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Day 3 .14 Not
.6667
6 significant

Day 4 .14 Not

.6667
6 significant

Day 5 1.0 Not

.0000
00 significant

Day 7 .71 Not

-.1667
1 significant

Day 7 Day 1 .00 Significant

2.0000*
0

Day 2 .00 Significant

1.5000*
2

Day 3 .07 Not

.8333
2 significant

Day 4 .07 Not

.8333
2 significant

Day 5 .71 Not

.1667
1 significant

Day 6 .71 Not

.1667
1 significant

*Calculation was performed at the 0.05 level of significance

Table 17. Table for Post Hoc Tests

Monitoring Time
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Multiple Comparisons

Dependent Variable: Hair Changes

LSD

95%

Confidence

(I) M Interval

onitorin (J) M Lowe

g onitoring Mean r

Ti Tim Difference Std. Boun Upper

me e (I-J) Error Sig. d Bound

Day 1 Day 2 -

-.5000 .44544 .271 1.412 .4124

Day 3 -

-1.1667* .44544 .014 2.079 -.2542

Day 4 -

-1.1667* .44544 .014 2.079 -.2542

Day 5 -

-1.8333* .44544 .000 2.745 -.9209

Day 6 -

-1.8333* .44544 .000 2.745 -.9209

Day 7 - -

-2.0000* .44544 .000 2.912 1.087

4 6

Day 2 Day 1 .5000 .44544 .271 -.412 1.412

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4 4

Day 3 -

-.6667 .44544 .146 1.579 .2458

Day 4 -

-.6667 .44544 .146 1.579 .2458

Day 5 -

-1.3333* .44544 .006 2.245 -.4209

Day 6 -

-1.3333* .44544 .006 2.245 -.4209

Day 7 -

-1.5000* .44544 .002 2.412 -.5876

Day 3 Day 1 .44544 .014 .2542 2.079

1.1667*
1

Day 2 .44544 .146 -.245 1.579

.6667
8 1

Day 4 .44544 1.00 -.912 .9124

.0000
0 4

Day 5 -

-.6667 .44544 .146 1.579 .2458

Day 6 -

-.6667 .44544 .146 1.579 .2458

Day 7 -.8333 .44544 .072 - .0791

1.745
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8

Day 4 Day 1 1.1667* .44544 .014 .2542 2.0791

Day 2 -.245
.6667 .44544 .146 1.5791
8

Day 3 1.00 -.912

.0000 .44544 .9124
0 4

Day 5 -

-.6667 .44544 .146 1.579 .2458

Day 6 -

-.6667 .44544 .146 1.579 .2458

Day 7 -

-.8333 .44544 .072 1.745 .0791

Day 5 Day 1 1.8333* .44544 .000 .9209 2.7458

Day 2 1.3333* .44544 .006 .4209 2.2458

Day 3 -.245
.6667 .44544 .146 1.5791
8

Day 4 -.245
.6667 .44544 .146 1.5791
8

Day 6 1.00 -.912

.0000 .44544 .9124
0 4

Day 7 -

-.1667 .44544 .711 1.079 .7458

Day 6 Day 1 1.8333* .44544 .000 .9209 2.7458

Day 2 1.3333* .44544 .006 .4209 2.2458

Day 3 -.245
.6667 .44544 .146 1.5791
8
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Day 4 -.245
.6667 .44544 .146 1.5791
8

Day 5 1.00 -.912

.0000 .44544 .9124
0 4

Day 7 -

-.1667 .44544 .711 1.079 .7458

Day 7 Day 1 1.087

2.0000* .44544 .000 2.9124
6

Day 2 1.5000* .44544 .002 .5876 2.4124

Day 3 -.079
.8333 .44544 .072 1.7458
1

Day 4 -.079
.8333 .44544 .072 1.7458
1

Day 5 -.745
.1667 .44544 .711 1.0791
8

Day 6 -.745
.1667 .44544 .711 1.0791
8

Based on observed means.

The error term is Mean Square(Error) = .595.

*. The mean difference is significant at the .05

level.

Table 18. Table for the Monitoring time in Multiple Comparison