ISOPROPYL AND ETHYL BASED ATSUETE HIGHLIGHTER INK: A

COMPARATIVE STUDY

RESEARCHERS

JC IZLEY B. BASSIG

ROSE ANN KATE L. GATAN

VINA G. MASIGAN

MARY ANGELA R. RAMOS

SHARMAINE D. CABACCAN

JENILYN P. MALLANAO

2023 – 2024

1
 ABSTRACT

This research investigates the properties and performance of highlighter inks formulated with atsuete

(annatto) pigment, utilizing isopropyl and ethyl alcohols as solvents. Atsuete, derived from the seeds

of the annatto tree, offers a natural and eco-friendly alternative for highlighter ink production. The

study aims to compare the chemical properties, effectiveness, longevity, and environmental impact

of isopropyl and ethyl-based atsuete highlighter inks. Grounded in principles of sustainable

chemistry and green engineering, the research explores the potential of atsuete highlighter inks to

reduce reliance on synthetic colorants and promote environmental sustainability. Experimental

procedures conducted within a chemistry laboratory include the preparation of atsuete extracts,

formulation of highlighter inks, and evaluation of ink properties such as color vibrancy, stability, ink

flow, and bleed-through on different paper types. Findings reveal comparable chemical properties

between isopropyl and ethyl-based atsuete highlighter inks, with minor variations in performance

observed. Both solvents demonstrate similar environmental impacts, prompting recommendations for

further research, long-term environmental assessment, education, awareness, and industry

collaboration to promote the development and adoption of sustainable highlighter ink formulations.

Overall, this study contributes to the advancement of eco-friendly stationery products and fosters a

deeper understanding of sustainable practices in ink manufacturing.

ACKNOWLEDGEMENT

The following people have the researchers' sincere gratitude for their encouragement, help, and

inspiration in making this study a success and finishing it.

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First and foremost to the All-Powerful God, who bestowed power, wisdom, and intelligence onto

the researchers to complete the study;

To the advisor, Mr. Garry Perez, for his valuable help and patience in providing them with the

guidance they needed to prepare for and finish the study,

Thank you to every respondent for your support. To all educators, I appreciate your concerns.

To our devoted family for their financial and moral support;

This piece of work is dedicated entirely to them.

TABLE OF CONTENTS

TITLE.................................................................................................................................I

ABSTRACT..........................................................................................................................II

ACKNOWLEDGEMENT.................................................................................................III

3
 CHAPTER I.......................................................................................................................6

INTRODUCTION..............................................................................................................7

STATEMENT OF THE PROBLEM............................................................................8

CONEPTUAL FRAMEWORK....................................................................................8

SIGNIFICANCE OF THE STUDY..............................................................................9

SCOPE AND DELIMITATIONS...............................................................................12

DEFINITION OF TERMS..........................................................................................13-14

CHAPTER II...................................................................................................................15

REVIEW OF RELATED LITERATURE AND STUDIES........................................15-17

CHAPTER III.............................................................................................................17

METHODOLOGY.............................................................................................................17

RESEARCH DESIGN....................................................................................................17

RESEARCH INSTRUMENT...............................................................................................17

LOCALE OF THE STUDY..............................................................................................18

MATERIAL AND PROCEDURES..............................................................................18-19

DATA ANALYSIS.............................................................................................................21

FLOW CHART...............................................................................................................21-22

4
 CHAPTER IV...................................................................................................................23

RESULTS AND DISCUSSION...........................................................................................23

CHAPTER V..................................................................................................................26

SUMMARY...................................................................................................................26

FINDINGS........................................................................................................................27

RECOMMENDATIONS.................................................................................................27-28

APPENDIX A.....................................................................................................29-30

APPENDIX B.....................................................................................................31-32

BIBLIOGRAPHY..........................................................................................................33

CHAPTER 1

INTRODUCTION

Highlighters have become indispensable tools in education and professional settings, aiding in the

emphasis and organization of textual information. Among the diverse range of highlighters

available, those incorporating atsuete (annatto) pigment have gained attention for their natural and

5
 eco-friendly composition. This study delves into the comparison of isopropyl and ethyl-based

atsuete highlighter inks, aiming to elucidate their chemical properties and evaluate their

performance in highlighting tasks.

Atsuete, derived from the seeds of the annatto tree (Bixa orellana), has been traditionally utilized

as a natural dye for various purposes due to its vibrant color and non-toxic nature. Its application

in highlighter ink formulations aligns with the growing demand for sustainable and

environmentally friendly alternatives in stationery products. Isopropyl and ethyl alcohols,

commonly used solvents in ink formulations, present themselves as suitable carriers for atsuete

pigments, each offering unique characteristics that may impact the final product.

The chemical composition of atsuete pigments contributes significantly to the overall properties

of the highlighter ink. Annatto seeds contain bixin and norbixin, which are carotenoid pigments

responsible for the characteristic orange to reddish hues. The stability and solubility of these

pigments in different solvents may vary, influencing the color intensity and longevity of the

highlighter ink. Investigating the interactions between atsuete pigments and isopropyl or ethyl

alcohol provides insights into the formulation's stability and shelf life.

Statement of the problem:

Generally, this study sought to investigate the properties of isopropyl and ethyl-based atsuete

highlighter inks, exploring their effectiveness, longevity.

Specifically, it sought to answer the following questions.:

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 1.) How do the chemical properties, including the solubility, stability, and color intensity, of

atsuete pigments in isopropyl-based highlighter ink compare to those in ethyl-based highlighter

ink?

2.) Is there a differences in the performance of isopropyl and ethyl-based atsuete highlighter inks

concerning factors such as ink flow, bleed-through on different paper types?

3.) How does the choice of solvent (isopropyl or ethyl alcohol) influence the overall

environmental impact of atsuete highlighter ink formulations, considering factors such as toxicity,

odor, and the renewable sourcing of the solvents?

THEORITICAL AND CONCEPTUAL FRAMEWORK

This study is grounded in the principles of sustainable chemistry and green engineering. By

utilizing natural dyes derived from Atsuete seeds, the research aims to reduce reliance on

synthetic colorants derived from petrochemicals, thereby mitigating environmental impact. The

choice of solvent (isopropyl vs. ethyl alcohol) is informed by their chemical properties, including

solubility, volatility, and toxicity. Theoretical frameworks such as Life Cycle Assessment (LCA)

and Green Chemistry Principles will guide the evaluation of environmental impacts and the

development of eco-friendly ink formulations. Additionally, color theory principles will inform

the assessment of color vibrancy and stability.

Conceptual Framework

INPUT PROCESS OUTPUT

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Figure 1. The paradigm of the study

The figure presents the paradigm of the study indicating Input, Process, and Output. The

framework's left side includes the study's Input, Atsuete, Isopropyl, Ethyl and Highlighter ink. On

the rmiddle, includes the process and Output of the study

Significance of the study

Ink Formulation Optimization:

This study can help optimize the formulation of highlighter inks by comparing the performance of

isopropyl and ethyl-based atsuete extracts. Understanding their differences in terms of color

vibrancy, stability , and other key properties can lead to the development of a more effective and

efficient highlighter ink.

Sustainability and Eco-friendliness:

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Investigating the use of atsuete extracts as a base for highlighter inks aligns with the growing

emphasis on sustainable and eco-friendly materials in various industries. If one of the extracts

proves to be more environmentally friendly, it could pave the way for the development of greener

highlighter products

Cultural and Traditional Application:

Atsuete is a traditional Filipino coloring agent with cultural significance. Understanding its

potential applications in modern products like highlighter inks not only bridges traditional

knowledge with contemporary technology but also promotes the preservation and utilization of

indigenous resources.

Health and Safety Considerations:

Isopropyl and ethyl alcohols are commonly used in various applications, but their safety and

potential health impacts should be thoroughly studied, especially when used in products like

highlighters. This research can shed light on any safety concerns associated with these ingredients,

contributing to the overall well-being of consumers.

Educational Implications:

The study can provide valuable information for educational purposes, especially in chemistry and

materials science. Students and educators can benefit from understanding the properties and

performance of different ink formulations, fostering a deeper appreciation for the science behind

everyday products.

Commercial Applications and Industry Innovation:

If one of the atsuete extracts demonstrates superior qualities as a highlighter ink, it could have

significant implications for the ink manufacturing industry. Companies may consider adopting the
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findings to enhance their product offerings, potentially leading to innovations in highlighter

technology.

Potential Cost Efficiency:

Analyzing the cost-effectiveness of isopropyl and ethyl-based atsuete highlighter inks can be

beneficial for manufacturers. If one formulation proves to be more cost-efficient without

compromising quality, it could result in economic advantages for the industry.

Market Differentiation:

The study's findings can offer a unique selling proposition for companies producing atsuete-based

highlighter inks. If there are distinct advantages in terms of color brilliance, longevity, or other

factors, it could position these products as premium or superior options in the

NULL HYPOTHESIS

There is no significant difference in the highlighting effectiveness between isopropyl-based and

ethyl-based atsuete highlighter inks in a comparative study.

ALTERNATIVE HYPOTHESES

There is a significant difference in the highlighting effectiveness between isopropyl-based and

ethyl-based atsuete highlighter inks in a comparative study.

SCOPE AND DELIMITATIONS

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The scope of this study encompasses the comparative analysis of isopropyl and ethyl based

atsuete (annatto) highlighter inks. It involves investigating their respective compositions,

properties, effectiveness as highlighter inks, and their impact on paper quality. The research will

delve into their chemical characteristics, such as solubility, stability, and color intensity.

Additionally, it will explore their compatibility with different types of paper, potential side effects

on paper texture, and longevity of highlighting effects. The study aims to provide insights into the

suitability of isopropyl and ethyl based atsuete highlighter inks for various applications, including

academic, artistic, and professional uses.

This study focuses specifically on atsuete-based highlighter inks formulated with isopropyl and

ethyl alcohol solvents. It does not include other types of highlighter inks or alternative solvents.

The research does not extend to the synthesis of atsuete pigment but rather assumes the

availability of commercial atsuete extract or dye for ink formulation. The investigation does not

encompass the environmental impact of the highlighter inks or their production processes.

Furthermore, the study does not address potential health hazards associated with prolonged

exposure to atsuete-based highlighter inks. The geographical scope of this study is not limited, but

the availability of materials and resources may affect the practical implementation of findings in

specific regions.

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Definition of terms

Isopropyl-based Atsuete Highlighter Ink - Refers to a highlighting ink formulation that utilizes

isopropyl alcohol as a solvent, incorporating atsuete (annatto) as a natural dye to produce color.

Ethyl-based Atsuete Highlighter Ink - Denotes a highlighting ink variant formulated with ethyl

alcohol as the solvent and atsuete (annatto) as the natural dye component.

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Atsuete (Annatto) - A natural dye derived from the seeds of the annatto plant (Bixa orellana),

known for its vibrant orange to red color.

Comparative Study - Refers to a systematic examination of isopropyl and ethyl-based atsuete

highlighter inks, aiming to identify and analyze differences in characteristics such as color

vibrancy, longevity, and overall performance.

Solvent - The liquid component of the ink responsible for dissolving and dispersing the atsuete

dye.

Color Vibrancy - Indicates the intensity and brilliance of the highlighted color produced by the

atsuete highlighter ink.

Longevity - Refers to the durability and persistence of the highlighted color over time. The

longevity of the ink's color is a critical factor in evaluating its practicality for long-term use.

Ink Formulation - Describes the specific combination and proportions of ingredients, including

atsuete, solvent, and any additional components, used in creating the atsuete highlighter ink.

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 CHAPTER II

REVIEW OF RELATED LITERATURE AND STUDIES

Annatto/Achiote Annatto , sometimes called roucou or achiote , is a derivative of the achiote trees

of tropical regions of the Americas, used to produce a yellow to orange food coloring and also as

a flavoring. Its scent is described as "slightly peppery with a hint of nutmeg " and flavor as

"slightly nutty, sweet and peppery".

Annatto coloring is produced from the reddish pericarp or pulp which surrounds the seed of the

achiote ( Bixa orellana L.). It is used as coloring in many cheeses (e.g., Cheddar , Gloucester

cheese , Red Leicester , Gouda and Brie ), margarine , butter , rice , custard powder, ice-cream ,

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and smoked fish . Although it is a natural food colorant, it has been linked to cases of food-related

allergies

Annatto is commonly found in Latin America and Caribbean cuisines as both a coloring agent and

for flavoring. Central and South American natives use the seeds to make a body paint and

lipstick . For this reason, the achiote is sometimes called the "lipstick- tree". Achiote originated in

South America and has spread in popularity to many parts of Asia. It is also grown in other

tropical or subtropical regions of the world, including Central America, Africa and Asia. The

heart-shaped fruit are brown or reddish brown at maturity, and are covered with short, stiff hairs.

When fully mature, the fruit split open, exposing the numerous dark red seeds. While the fruit

itself is not edible, the orange-red pulp that covers the seed is used to produce a yellow to orange

commercial food coloring. Achiote dye is prepared by grinding seeds or simmering the seeds in

water or oil.

For centuries, natural materials including berries, bark, and leaf extracts have been used to make

ink. When combined with other substances, they have been used as basic ingredients to generate a

variety of colors as well as ink, dye, and paint. Due to its ability to generate ink in many shades—

yellow, green, brown, or black—tea leaves were employed to manufacture ink according to early

accounts (Piero et al., 2005).

The development of writing ink has advanced significantly over the years in line with

technological advancements to satisfy the needs of both the public and private sectors.

Chalkboards began to give way to whiteboards with non- permanent inks in classrooms. This was

in line with the worries about the health hazards that chalk dusts pose. It also cleans up easier and

15
 has a more appealing appearance.Solvents derived from petroleum and chemicals, which are non-

renewable synthetic resources, are frequently used to make inks today. These materials are

hazardous to both the environment and its users. Because of the ink's solvents and pigments, such

p-anisidine, unprotected ink contact can result in excruciating headaches, skin irritation, or even

harm to the nervous system (Basri et al., 2021) Despite their many advantages, these non-

permanent inks from Whiteboard have severe drawbacks (Brown, n.d). In addition, Synthetic inks

used to make whiteboard markers are particularly hazardous since the non-permanent ink releases

Xylene, a petroleum toxin that can have adverse effects on health, including skin headaches,

nausea, dizziness, and irritability (Jolly and Wais, 2018).

Since natural dyes are created using cleaner procedures, are less toxic, non-carcinogenic, and

naturally biodegradable, using them is an ecologically benign option (Gonçalves et al., 2021).

Two main components such as Gumamela and Annatto seeds are the best examples in making

alternative marker ink for it does not contain hazardous chemicals and solvents. Additionally, the

increased focus on sustainability worldwide has led to the use of natural dyes in a variety of

applications, including writing (Chew, 2021), food packaging (Hoffmann et al., 2022), and food

coloring (Caro et al., 2012).

Annatto (Bixa orellana L.), which is made naturally by macerating the pericarp of achiote seeds, is

one of the first known natural dyes. When grown in tropical climates, this plant, which is native to

South and Central America (Boguniewicz-Zablocka et al., 2020), produces seeds all year round

(Singh and Bharati, 2014). The industry has recently shown more interest in annatto because of its

possible use as a dye. High biodegradability, little toxicity, and excellent environmental

compatibility characterize annatto dye (Shahid-Ul-Islam et al., 2016).

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 CHAPTER III

This chapter presents the different procedures and tests that we conducted in this study,

including the research design, the locale of the study, materials and procedures, data collection,

computation of mortality rates, statistical analysis, and expected output.

RESEARCH DESIGN

The study used a quantitative experimental research design to evaluate the Isopropyl and

ehtyl based atsuete highlighter ink. Stratified sampling was used to choose a respondents.

RESEARCH INSTRUMENT

The research instrument consists of a structured questionnaire designed to collect quantitative data

from participants. The questionnaire includes items related to various aspects of the highlighter

inks, such as color vibrancy, stability, ink flow, and environmental impact. Participants are asked
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to rate their agreement with each statement using a 5-point Likert scale, ranging from "Strongly

Disagree" to "Strongly Agree."

LOCALE OF THE STUDY

Preparation of materials, storing, and experimentation was conducted inside a chemistry

laboratory of Rodolfo B. Albano Stand Alone Senior High School at Barangay Catabayungan,

Cabagan, Isabela from Ferbruary 28, 2024.

Materials

Atsuete powder (Annatto seeds)

Isopropyl alcohol

Ethyl alcohol

Glass containers

White paper

Gloves

Hairnet

Spoon

Labgown

Beaker

Highlighter ( with empty ink)

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Procedure

Prepare the Atsuete Extract:

a. Measure out a suitable amount of Atsuete powder, depending on how much ink you want to

make. Around 4 tablespoon of atsuete powder is a good starting point.

b. Place the Atsuete powder into the beaker

c. Pour Isopropyl alcohol into the beaker until the atsuete powder are fully submerged.

Alternatively, you can use Ethyl alcohol or a mixture of both Isopropyl and Ethyl alcohol for

different properties in the final ink (such as drying time and color intensity).

Extract the Color

a. Put it into the beaker and stir it gently to ensure the powder are well covered with the alcohol.

b. Let the beaker sit in a cool place for at least 5 minutes. This allows the alcohol to extract the

color from the Atsuete powder.

Strain the Mixture

a. After 5 minutes, place it over a small bowl.

b. Carefully pour the contents of the beaker, allowing the liquid to filter into the bowl.

Transfer to Container

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 a. Using a funnel if necessary, carefully pour the cooled ink into a dropper bottle or the refill of a

highlighter pen.

b. Seal the bottle or pen tightly to prevent evaporation or spillage.

Testing and Adjustments

a. Before using the ink, test it on a scrap piece of paper to ensure it has the desired color intensity

and consistency.

b. If the color is too light, you can repeat the heating process to further concentrate the ink.

Conversely, if it's too dark, you can dilute it with more alcohol.

Labeling and Storage

a. Label the container, treatment 1 and 2.

b. Store the highlighter ink in a cool, dry place away from direct sunlight to maintain its quality

and shelf life.

DATA ANALYSIS

Data collected from the experiments will be analyzed using appropriate statistical methods

to compare the properties and performance of isopropyl and ethyl-based atsuete highlighter inks.

Descriptive statistics such as mean, standard deviation, and graphical representations will be used

20
to summarize the data. Inferential statistics, including t-tests and analysis of variance (ANOVA),

will be employed to determine significant differences between the two ink formulations.

FLOW CHART

Research Design

Locale of the study

Materials & Procedures

Preparation of Atsuete Extract

Extract the Color

Strain the Mixture

Transfer to Container

21
Testing and Adjustments

Labeling and Storage

Data Analysis

Statistical Analysis

Expected Output

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 CHAPTER IV

RESULTS AND DISCUSSION

This chapter presents the data gathered with the corresponding statistical analysis and

interpretation of data collected from the experiments and observation of researchers. The following

are the results of each treatment and statistical analysis.

Table 1. Demographic Profile

Demographic Frequency Percentile

Sex

Female 11 73.3%

Male 4 26.7%

Total 15 100%

Table 1 shows that the demographic profile of the respondents. It is clear that the majority are

female with a total of 12 or 73.3% .

Table 2.

Demographic Frequency Percentile

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 Age

15 1 6.67%

16 9 60%

17 5 33.33 %

Table 2 shows the demographic profile of the respondents. It is clear that the majority age of
respondents are 16 with a total of 9 or 60% .

Table 3.

DP R1 R2 R3 R R R6 R R8 R9 R1 R R1 R R R Total
4 5 7 0 11 2 13 1 1
4 5

Sex F F F F F F F F M M M M F F F F=11

M=4

Age 16 17 17 16 16 16 16 17 17 16 15 17 16 1 1 17=5
6 6
16=9

15=1

Statement Mean

1 2 3 3 3 3 4 3 2 1 3 1 3 3 3 3 2.67%

2 5 5 5 5 5 4 5 4 5 5 5 5 5 5 5 4.87%

3 4 4 4 4 4 3 4 4 5 3 5 5 3 4 5 4.7%

4 4 5 4 4 4 4 4 4 5 4 5 1 4 4 5 4.7%

5 4 1 2 2 3 2 4 3 3 3 4 4 2 2 2 2.73%

6 4 5 3 2 2 3 3 3 5 5 5 5 5 4 4 3.9%

7 3 3 2 4 4 3 3 5 2 2 2 1 2 2 1 2.6%

8 3 2 2 2 2 3 3 5 1 2 1 1 1 2 1 2.7%

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9 3 3 3 4 3 2 2 3 2 1 2 2 2 2 2 2.4%

10 5 5 5 2 2 5 5 3 5 5 5 5 5 5 5 4.5%

11 4 4 4 3 3 4 2 4 2 3 2 2 3 3 3 3.7%

12 5 5 5 3 5 4 5 5 5 5 5 4 5 5 5 4.73%

13 2 2 3 2 3 2 2 2 2 3 2 4 3 3 3 2.53%

14 5 3 3 2 5 4 5 5 5 4 5 5 5 3 3 4.30%

15 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1%

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 CHAPTER V

SUMMARY, FINDINGS, RECOMMENDATIONS

Summary:

The study aimed to compare the properties and performance of isopropyl and ethyl-based atsuete

(annatto) highlighter inks, focusing on their effectiveness, longevity, and environmental impact.

Atsuete, derived from the seeds of the annatto tree, was chosen for its natural and eco-friendly

composition. Isopropyl and ethyl alcohols were selected as solvents due to their common usage in

ink formulations. The research was grounded in principles of sustainable chemistry and green

engineering, aiming to provide insights into sustainable ink formulation.

The study utilized a quantitative experimental research design, conducting experiments within a

chemistry laboratory. Atsuete extracts were prepared using both isopropyl and ethyl alcohols, and

highlighter inks were formulated based on these extracts. Various tests and procedures were

conducted to evaluate the properties of the highlighter inks, including color vibrancy, stability, ink

flow, and bleed-through the paper.

Findings:
26
 1. Chemical Properties: Both isopropyl and ethyl-based atsuete highlighter inks demonstrated

similar chemical properties in terms of solubility, stability, and color intensity of atsuete pigments.

The choice of solvent did not significantly affect these properties.

2. Performance: The performance of isopropyl and ethyl-based atsuete highlighter inks in terms of

ink flow and bleed-through the paper that is showed minor variations. Isopropyl-based inks exhibited

slightly better ink flow, while ethyl-based inks showed slightly less bleed-through on a certain paper.

3. Environmental Impact: Both solvents, isopropyl and ethyl alcohol, showed comparable

environmental impacts in terms of toxicity, odor, and renewable sourcing. However, further analysis

is needed to evaluate their long-term environmental effects.

Recommendations:

1. Further Research: Future studies could explore additional solvent options and their impact on the

properties of atsuete highlighter inks. Investigating alternative natural dyes and their compatibility

with different solvents could also contribute to the development of eco-friendly highlighter products.

2. Long-Term Environmental Assessment: Conducting a comprehensive life cycle assessment (LCA)

of atsuete highlighter inks, considering factors such as production, usage, and disposal, would

provide a clearer understanding of their overall environmental impact. This could guide the

development of more sustainable ink formulations.

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3. Education and Awareness: Promoting awareness about the benefits of using eco-friendly

stationery products, such as atsuete highlighter inks, could encourage consumers to make

environmentally conscious choices. Educational initiatives in schools and workplaces could highlight

the importance of sustainability in everyday practices.

4. Industry Collaboration: Collaboration between ink manufacturers, environmental experts, and

regulatory bodies could facilitate the adoption of sustainable practices in the ink manufacturing

industry. Developing industry standards for eco-friendly ink formulations and promoting green

certification could drive innovation towards more sustainable products.

APPENDIX A

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 The researchers measure out a suitable amount of Atsuete powder and Pour Isopropyl alcohol

into the beaker until the atsuete powder are fully submerged.

29
APPENDIX B

30
 The researchers measure out a suitable amount of Atsuete powder and Pour Ethyl alcohol into

the beaker until the atsuete powder are fully submerged.

31
The researchers labeled the container, treatment 1 and 2.

32
The researchers use the ink before testing it on a scrap piece of paper to ensure it has the desired

color intensity and consistency. The Final results is that the treatment 2 is more effective than the

treatme

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