Laporan Skripsi-revisi

THESIS
TEXTURE OPTIMIZATION OF FUNCTIONAL COOKIES
WITH THE ADDITION OF CHIA (Salvia hispanica L.) FLOUR
Written as partial fulfillment of the academic requirements
to obtain the degree of Sarjana Teknologi Pertanian Strata Satu
By :
NAME : ANGGADA PUTRA
NPM : 03420110004
FOOD TECHNOLOGY DEPARTMENT
FACULTY OF SCIENCE AND TECHNOLOGY
UNIVERSITAS PELITA HARAPAN
2015

STATEMENT OF THESIS AUTHENTICITY
I, a student of Food Technology Department, Faculty of Science and Technology,
Universitas Pelita Harapan,
Name : Anggada Putra
Student Id. Number : 03420110004
Department : Food Technology
hereby declare that my thesis, entitled “TEXTURE OPTIMIZATION OF
FUNCTIONAL COOKIES WITH THE ADDITION OF CHIA (Salvia
hispanica L.) FLOUR” is:
1) An original piece of work, written and completed on my own, based on lecture
notes, data observation, reference books, journals, and other sources as listed
on the work cited section.
2) Not a duplication of other writings that have been published or used for
obtaining the degree of Sarjana in any Universities, except for passages that
include information on respective references.
3) Not a translation of other works.
I understand that if my statement above is proven untrue, this Thesis will be
cancelled.
Tangerang, July 14th 2015
(ANGGADA PUTRA)

APPROVAL BY THESIS SUPERVISORS
TEXTURE OPTIMIZATION OF FUNCTIONAL COOKIES WITH
THE ADDITION OF CHIA (Salvia hispanica L.) FLOUR
Written by :
has been examined in the thesis examination for obtaining the degree of Sarjana
Teknologi Pertanian Strata Satu in the Food Technology Department, Faculty of
Science and Technology, Universitas Pelita Harapan, Karawaci - Tangerang,
Banten, and has been approved by the thesis supervisors.
Tangerang, July 14th, 2015
Approved by:
Supervisor Co-Supervisor
(Prof. Dr. C. Hanny Wijaya) (Jeremia M. Halim, MP)
Acknowledged by:
Head of Department Dean
(Julia Ratna Wijaya, MAppSc) (Prof. Dr. Manlian Ronald. A., ST, MT.)

APPROVAL BY THESIS EXAMINATION COMMITTEE
We the undersigned, certify that a thesis defense has been held on June 30th, 2015,
as partial fulfillment of the academic requirements to obtain the degree of Sarjana
Teknologi Pertanian Strata Satu in Food Technology Department, Faculty of
Science and Technology, Universitas Pelita Harapan, for the student:
Faculty : Science and Technology
with the following title “TEXTURE OPTIMIZATION OF FUNCTIONAL
COOKIES WITH THE ADDITION OF CHIA (Salvia hispanica L.) Flour” and
that the thesis was successfully defended, henceforth approved by the examination
committee.
Examiners Signature
1. Dr. Hardoko Head of Examiners
2. Prof. Dr. C. Hanny Wijaya Member
3. Lisa A. Yakhin, M. Eng Member

v
ABSTRACT
Anggada Putra (03420110004)
TEXTURE OPTIMIZATION OF FUNCTIONAL COOKIES WITH THE
ADDITION OF CHIA (Salvia hispanica L.) FLOUR
(xiii + 51 pages: 14 figures, 14 tables, and 24 appendices)
The addition of chia flour towards the formulation of baked products, including
pound cake and bread shows that it can decrease the specific volume and texture
parameter of these products, respectively. This research’s objective is to obtain
the best formulation of chia flour and hydrogenated vegetable fat (HVF) to
produce cookie with optimum texture quality based on sensory evaluation and
physico-chemical analysis. Different contents of chia flour (0 - 30 g) and HVF (33
- 55 g) were added to the cookie mix using Response Surface Methodology (RSM)
based on 22 central composite rotational design (CCRD). Chia flour and HVF
significantly affects the acceptability of cookies produced in terms texture.
Physicochemical parameters, including hardness, fracturability, spread ratio, and
moisture content were also affected. The formula to obtain cookie with optimum
texture is 20.45 g of chia flour and 55.00 g of HVF. The nutrition content of
optimum chia cookie is 8.75 % protein, 27.81 % fat, 1.64 % omega-3 fatty acid,
1.18 % ash, 58.46 % carbohydrate, 9.16 % total dietary fiber, and 3.80 %
moisture content. Based on the nutrition content, optimum chia cookie can be
claimed as a good source of fiber and high source of α-linolenic acid (ALA)
according to FDA.
Keyword: Chia, Fat, Cookie, Omega-3, Dietary Fiber, Texture.
References: 47 (1992 - 2015)

vi
ACKNOWLEDGEMENTS
Praise the Lord for His blessing during the research and the making of
thesis report that this report can be finished well. The completion of thesis report
entitled “TEXTURE OPTIMIZATION OF FUNCTIONAL COOKIES
WITH THE ADDITION OF CHIA (Salvia hispanica L.) FLOUR” is one of
the requirements to achieve bachelor degree in Sarjana Strata Satu Teknologi
Pangan. Author realizes that the report could not be finished without the guidance,
assistance, prayer, and support from many parties. Hence, author would like to
express gratitude to those who help and support author during this time, including:
1. Prof. Dr. C. Hanny Wijaya as the thesis supervisor for the guidance, time,
and support during research and thesis writing.
2. Mr. Jeremia Halim as thesis co-supervisor for the guidance, advice, and
support during the research and thesis report writing.
3. Ms. Julia Ratna Wijaya, MAppSc as the head of Food Technology
Department who gave chance to author to complete this thesis.
4. Beloved father and mother for the endless love, prayers, and supports for
author. And also for brothers who supports during the most difficult time.
5. Mr. Hendra, Mr. Aji, Mr. Darius and Mrs. Meri as laboratory assistant for
the help and support during research in laboratories.
6. Anastasia Stephanie, Nicholas Adams, Gabriel Eugenie, Amanda Inggita,
and Alvin Kusuma who accompanied, helped, and being cooperative with
the author during work in laboratories.
7. Edison Sutiono, Marvin Setiawan, Natanael Leon, and Aditya Febrian for

vii
constant support during the research and the completion of report.
8. All members of C Class of Food Technology 2011 Universitas Pelita
Harapan who are not be mentioned above.
9. All friends and close relatives that have not been mentioned but provided
help and support for author during internship and report completion.
Author realized author might have done mistakes during research and
report completion. Therefore, the author would welcome any critics and
suggestions upon this report. Finally, author hopes that this report would be useful
for the reader.
Tangerang, July 14th, 2015
Author

viii
TABLE OF CONTENTS
COVER page
STATEMENT OF THESIS AUTHENTICITY
APPROVAL BY THESIS SUPERVISORS
APPROVAL BY THESIS EXAMINATION COMMITTEE
ABSTRACT ............................................................................................................v
ACKNOWLEDGEMENTS..................................................................................vi
TABLE OF CONTENTS.....................................................................................viii
LIST OF TABLES ................................................................................................xi
LIST OF FIGURES ............................................................................................. xii
LIST OF APPENDICES......................................................................................xiii
CHAPTER I INTRODUCTION ...........................................................................1
1.1 Background ....................................................................................................1
1.2 Research Problem...........................................................................................2
1.3 Objectives.......................................................................................................2
1.3.1 General Objectives ..................................................................................2
1.3.2 Specific Objectives..................................................................................2
CHAPTER II LITERATURE REVIEW.............................................................4
2.1 Overview of Cookie .......................................................................................4
2.1.1 Pressed Cookies.......................................................................................4
2.2 Cookie Ingredients .........................................................................................4
2.2.1 Flour ........................................................................................................4
2.2.2 Sugar........................................................................................................5
2.2.3 Fat............................................................................................................5
2.2.4 Egg ..........................................................................................................6
2.2.5 Leavening Agent .....................................................................................6
2.2.6 Water .......................................................................................................7
2.2.7 Additional Ingredients.............................................................................7

ix
2.3 Cookie Processing..........................................................................................8
2.4 Chia ................................................................................................................9
2.4.1 Nutritional Value of Chia Seed .............................................................10
2.4.2 Utilization of Chia Seed in Food...........................................................12
2.5 Omega-3 Fatty Acid.....................................................................................13
2.6 Dietary Fiber ................................................................................................15
2.7 Response Surface Methodology...................................................................16
2.7.1 Central Composite Design ....................................................................16
CHAPTER III RESEARCH METHODOLOGY .............................................18
3.1 Materials and Equipments............................................................................18
3.2 Research Procedure......................................................................................18
3.2.1 Preliminary Research ............................................................................18
3.2.2 Main Research.......................................................................................20
3.3 Experimental Design....................................................................................23
3.3.1 Preliminary Research ............................................................................23
3.3.2 Main Research.......................................................................................23
3.4 Analysis Procedure.......................................................................................24
3.4.1 Physico-chemical Analysis ...................................................................24
3.4.2 Sensory Evaluation................................................................................28
CHAPTER IV RESULT AND DISCUSSION ...................................................30
4.1 Overview ......................................................................................................30
4.2 Nutritional Content of Chia Flour ................................................................30
4.3 Determination of Mixing Method ................................................................31
4.4 Formula Optimization of Cookies................................................................33
4.4.1 Optimization of Sensory Evaluation Result..........................................34
4.4.2 Optimization of Physical Analysis Result.............................................36
4.4.3 Optimal Formula Recommendation......................................................45
4.5 Product Verification.....................................................................................46
4.6 Product Comparison.....................................................................................47
4.6.1 Sensory Evaluation Result ....................................................................47

x
4.6.2 Physico-chemical Analysis ...................................................................48
4.6.3 Nutrition Composition...........................................................................49
CHAPTER V CONCLUSION AND SUGGESTION .......................................51
5.1 Conclusion....................................................................................................51
5.2 Suggestion....................................................................................................51
BIBLIOGRAPHY ................................................................................................52
APPENDICES ......................................................................................................56

xi
LIST OF TABLES
page
Table 2.1 Composition of chia seed per 100 g......................................................11
Table 3.1 Basic formulation for cookies ................................................................19
Table 3.2 Concentration composition of cookies formula .....................................22
Table 3.3 Formulation Combination Model...........................................................22
Table 3.4 Optimization cookies formula goal and importance ..............................22
Table 4.1 Nutritional Content of Chia Flour..........................................................30
Table 4.2 Hardness Value of Cookie Made With Different Mixing Methods .......33
Table 4.3 Statistical Model for Each Responses....................................................34
Table 4.4 Criteria and Goals of Each Responses for Optimal Cookie Formulation
................................................................................................................................46
Table 4.5 The Optimal Formula of Cookie Formulated With Chia Flour .............46
Table 4.6 Comparison Between Prediction Value and The Actual Result ............47
Table 4.7 Result of Sensory Evaluation Test .........................................................47
Table 4.8 Result of Physico-chemical Analysis.....................................................48
Table 4.9 Nutritional Content of Optimum Chia Cookie and Control Cookie ......49

xii
LIST OF FIGURES
page
Figure 2.1 Salvia hispanica L. ...............................................................................10
Figure 3.1 Flowchart of the cookies making..........................................................21
Figure 4.1 Result of Paired Comparison Test ........................................................31
Figure 4.2 Result of Paired Preference Test...........................................................32
Figure 4.3 3-D Graph for Texture Response Based on Hedonic Test....................35
Figure 4.4 Contour Graph for Texture Response Based on Hedonic Test.............36
Figure 4.5 3-D Graph for Hardness Value .............................................................37
Figure 4.6 Contour Graph for Hardness Value ......................................................37
Figure 4.7 3-D Graph for Fracturability Value ......................................................40
Figure 4.8 Contour Graph for Fracturability Value ...............................................39
Figure 4.9 3-D Graph for Spread Ratio..................................................................41
Figure 4.10 Contour Graph for Spread Ratio .........................................................42
Figure 4.11 3-D Graph for Moisture Content ........................................................45
Figure 4.12 Contour Graph for Moisture Content..................................................44

xiii
LIST OF APPENDICES
page
Appendix A. Identification Result ...................................................................... A-1
Appendix B. Questionnaire of Comparison Test .................................................B-1
Appendix C. Result of Comparison Test ............................................................ C-1
Appendix D. Statistical Analysis of Comparison Test........................................ D-1
Appendix E. Result of Texture Analysis in Preliminary Analysis.......................E-1
Appendix F. Questionnaire of Hedonic Test........................................................F-1
Appendix G. Result of Hedonic Test ................................................................. G-1
Appendix H. Analysis of Texture Response ....................................................... H-1
Appendix I. Result of Hardness and Fracturability Analysis................................I-1
Appendix J. Analysis of Hardness Response ........................................................J-1
Appendix K. Analysis of Fracturability Response..............................................K-1
Appendix L. Data of Spread Ratio .......................................................................L-1
Appendix M. Analysis of Spread Ratio Response ..............................................M-1
Appendix N. Data of Moisture Content ..............................................................N-1
Appendix O. Analysis of Moisture Content Response .......................................O-1
Appendix P. Questionnaire for Verification Test.................................................P-1
Appendix Q. Result of Hedonic Test in Verification Test..................................Q-1
Appendix R. Result of Physicochemical Analysis in Verification Test ..............R-1
Appendix S. Questionnaire for Product Comparison...........................................S-1
Appendix T. Result of Hedonic Test in Product Comparison..............................T-1
Appendix U. T-Test Analysis for Hedonic Test in Product Comparison ........... U-1
Appendix V. Physicochemical Analysis Result in Product Comparison............ V-1
Appendix W. Analysis for Physicochemical Test in Product Comparison........ W-1
Appendix X. Result of Proximate Analysis ........................................................ X-1

1
CHAPTER I
INTRODUCTION
1.1 Background
Biscuits and cookies are food products which have a very significant part
in the food industry in most countries (Manley, 2011). The reasons for this are due
to their relatively long shelf life, great convenience as food products, good value
for money, and human affinity, especially children, towards products which
contains sugar. However, according to Manley (2011), and Bassinello et al.
(2011) cookie lacks dietary fiber and essential fatty acids.
The interest on research, development and commercialization of functional
food ingredients, nutraceuticals and dietary supplements have been growing
around the globe (McManus et al., 2011). Chia (Salvia hispanica L.) is an annual
plant which grows mainly in South America, known to be rich in dietary fiber and
omega-3 fatty acid. For this reasons, it is expected that chia seeds can be utilized
as an ingredient for functional food, which in this case acts as a substitute of
wheat flour in formulation of cookie (Reyes Caudillo et al., 2007). Research done
by Pizarro et al. (2013) and Steffolani et al. (2015) shows that chia flour can be
added into the formulation of pound cake and bread, respectively. However, the
result of both experiments showed that the addition of chia can decrease the
specific volume of the final product and texture parameter based on sensory
evaluation result, but an increase in the amount of hydrogenated vegetable fat can
help overcome the problem. Fat is important for texture parameter, due to its
ability to prevent excessive gluten development by preventing water from reacting

2
with glutenin and gliadin to form gluten. Optimization of the amount of chia flour
and hydrogenated vegetable fat added is expected to produce cookies with
acceptable texture comparable to most cookies. However, because of the usage of
new ingredient in the making of cookies, the method used to make the cookie
need to be adjusted (Novianty, 2015). Therefore it is important to determine the
best mixing method.
1.2 ResearchProblem
The addition of chia flour in wheat flour-based product such as cake and
bread is known to cause a decrease in its specific volume and lower texture
acceptability compared to the control. The mixing method of cookie have to be
adjusted as well, as new ingredient is incorporated in the formulation. The correct
amount of hydrogenated vegetable fat is known to be able to increase the specific
volume and texture characteristics of the wheat flour-based product. Hence,
finding the correct formulation of chia seeds flour and hydrogenated vegetable fat
for use in cookie making is expected to overcome this problem.
1.3 Objectives
1.3.1 General Objectives
The general objective of this research was to produce cookies with the
optimal texture using chia flour and hydrogenated vegetable fat.
1.3.2 Specific Objectives
The specific objectives of this research were:
1. To determine the best mixing method of cookie made with the addition of chia
flour.

3
2. To study the effects of incorporating different amount of chia flour and
hydrogenated vegetable fat towards the texture characteristics of cookie.
3. To evaluate the acceptability of texture parameter of cookie made with the
addition of chia flour based on sensory evaluation.

4
CHAPTER II
LITERATURE REVIEW
2.1 Overview of Cookie
Cookie is a name which originally comes from the dutch word Koekje,
which basically means a small cake. Most of the components found in cookie is
the same as cake, but it is different in that it contains lower water content and
higher sugar and fat content. Based on its dough or batter’s fluidity, cookies can
be classified into bar, dropped, pressed, molded, rolled, and icebox/refrigerator
cookie (Brown, 2011; Manning, 2011).
2.1.1 PressedCookies
Pressed cookies have a flour mixture which is viscous enough to be stuffed
into a pastry bag or cookie press. Examples of cookies of this type are ladyfingers
and coconut macaroons (Brown, 2011).
2.2 Cookie Ingredients
Main ingredients used in making cookies are flour, sugar, fat, egg,
leavening agent, and water. Additional ingredients might be added depending on
the desired final product (Sumnu and Sahin, 2008).
2.2.1 Flour
Wheat flour is the most used flour in cookie making. In cookie, gluten
formation is avoided, so soft wheat flour is generally preferred than heavy wheat
flour. The reason is because the desired texture for cookie is crispy and soft, and
heavy wheat flour with high protein content causes the the texture of the cookie to

5
become hard and thick. Moreover, higher protein concentration have been
correlated with reduced diameter in cookie making as well, although other factors
are suspected to play a role as well. Rate of spreading of cookie dough is found to
be faster as well in cookies made from soft wheat flour. This is because the
amount of soluble starch in soft wheat flour is low, and causes the dough to be
less viscous which in turn increases its spreading rate (Brown, 2011; Sumnu and
Sahin, 2008).
2.2.2 Sugar
Sugar is added into cookie to add sweetness, acts as a tenderizing agent,
and affects spread. It was observed using a farinograph, that an increase in sugar
concentration reduces the consistency and cohesion of cookie dough. As a
hardening agent, sugar crystallizes as the cookie dough cools down after baking
and thereby makes the product crispy. However, it can act as softener in moderate
amounts, due to sugar’s ability to retain water. Sugar also makes the final product
to become fragile, because it controls hydration and tends to disperse starch and
protein molecules, which in turn prevents the formation of a continuous mass
(Sumnu and Sahin, 2008).
2.2.3 Fat
Fat is an essential ingredient in cookie, as its addition influences the
texture and taste of cookies, which makes the cookie crispier because this allows
the dough to spread as it cooks on the cookie sheet (Jacob and Leelavathi, 2007).
Fat has the function of preventing water from forming gluten network with
glutenin and gliadin protein from wheat flour, thereby making the dough less
resistant to mixing. Formation of gluten makes cookie dough more elastic and

6
resistant to mixing, making the end product hard. Fat also entraps air particles in
the dough, thereby increasing the total volume and lower the density of cookie
produced. Additionally, during baking the fat melts and helps the cookie dough
spread in the cooking sheet. This makes cookie have its characteristic soft and
crispy texture (Jacob and Leelavathi, 2007).
The source of fat used for cookie varies, but vegetable fat is generally used
more compared to animal fat, due to the origin of these materials, their quality is
variable and can cause issues to the final product. Also, as manufacturers wanted
to sell their product to all consumers, including the vegetarians, vegetable fat
becomes preferrable (Sumnu and Sahin, 2008; Manley, 2011).
2.2.4 Egg
The addition of egg into cookie dough helps in puffing, emulsifying the
dough, due to the presence of lecithin, which is an emulsifier. Emulsifier helps
entrap air particles and allow water to mix with fat, preventing it from reacting
with glutenin and gliadin to form gluten (Jacob and Leelavathi, 2007). This results
in a creamier and smoother texture in cookies. Presence of fat in egg contributes
to the final taste and add the total fat in cookies as well. The egg white component
contributes toward the structure or shape of the cookie dough, due to the drying
effect it has (Sumnu and sahin, 2008; Manley, 2011).
2.2.5 Leavening Agent
The presence of leavening agent causes the cookie dough to rise, which in
turn causes the final product to have greater volume and superior texture.
Leavening agent may be of physical, biological, or chemical in nature (Sumnu and
Sahin, 2008). Physical leavening agents are air and steam, which is done through

7
mechanical action, such as creaming, beating, or heating. Biological leavening
agents are yeasts and bacteria. Carbon dioxide produced by these microorganisms
through fermentation causes the dough to rise. Chemical leavening agents are
baking powder and baking soda, which yields carbon dioxide when it is heated or
mixed with acid. Sodium bicarbonate is the most commonly used chemical
leavening agent (Brown, 2011).
2.2.6 Water
Water is used in a very small amount in cookie making, usually only about
3 - 4 % of the whole cookie formulation. It’s purpose is generally to dissolve dry
ingredients and provide steam for leavening during baking. Too much water is
unfavorable, because it will induce the formation of gluten in the dough, causing
the texture of the cookie to be less crispy and more hard (Brown, 2011).
2.2.7 Additional Ingredients
Nonfat dry milk is sometimes added into cookies to give subtle flavor and
textural improvements and to aid surface coloring (Brown, 2011). Reducing agent
can also be added as an additive to increase cookie spread, by reducing disulfide
bonds to the sulfhydryl group, thereby decreasing dough stability. Cornstarch
sometimes is used in cookie as anticaking agent, drying agent, formulation aid,
processing aid, surface-finishing agent. Other food additives which is used in
cookies are potassium sorbate as preservative, soybean lecithin as emulsifier, and
whey as a binder ot extender to provide a uniform texture (Sumnu and Sahin,
2008).

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2.3 Cookie Processing
The processing of cookie is basically comprised of four steps, which are
dough making, processing and shaping, baking, cooling and packaging. The first
step of the process is dough making. In this step, all ingredients used to make
cookies are mixed together to form a dough. During mixing, the protein in flour
contacts with water and swell. If the mixing is continued, the protein containing
water will form a three-dimensional gluten network. Sugar should be dissolved by
the water in this process, as otherwise the sugar crystal will caramelize during
baking, causing brown spots. Dough temperature is also important in terms of the
fat used. At high temperatures, the fat will melt and the dough becomes fatty. The
dough should be maintained to have plastic-like texture (Sumnu and Sahin,
2008). Depending on the desired cookie characteristic, cookie processing can
categorized into one step and two step mixing (Hui, 2006). In two step mixing, the
fat and sugar are mixed first, followed by egg, and finally the flour and other dry
ingredients. Mixing cookie ingredients in two stages can minimize the
development of gluten in the flour, which is desirable in cookie dough. By
mixing the cookie ingredient in two stages, the fat will be more evenly dispersed
around the flour particles, and therefore rendering them less available to water and
preventing gluten development. In one-stage mixing, all the ingredients are mixed
at once. However, in this step the baker has less control over the mixing, and
therefore, the gluten development as well. However, it can be used when over-
mixing is not a serious problem, for example in the making of chewy cookies.
The second step is processing and shaping. In this step the doughs are
shaped to the desired form. The machines used for processing varies depending on

9
the hardness of the dough. Very soft dough needs to be deposited directly onto the
steel oven band invariably. For soft and usually sticky dough, it needs to be
extruded and cut using wire-cut machine. Dough which are stiff should be
extruded on a bar machine or rout press machine (Sumnu and Sahin, 2008).
The third step is baking. In this process, the dough of the cookies should
be placed far enough apart on the baking sheet to prevent them from touching
together during baking. Cool cooking sheets should be places as well on the
baking sheet to prevent overspreading (Brown, 2011). Temperatures ranging
between 163 - 191 oC are usually used in baking of cookies. The next and last step
after baking is cooling and packaging. After baking, the cookies should be placed
on a cloth band and cooled down in room temperature. Placing the recently baked
cookie on a temperature which is too cold might cause cracking on the surface.
The cookies can be placed on the packaging after it is sufficiently cooled down
(Sumnu and Sahin, 2008).
2.4 Chia
Chia is a plant native to the central valleys of Mexico and northern
Guatemala. The word “chia” is a Spanish adaptation of the word chian or chien in
its plural form, meaning “oily”, which comes from Nahuatl, the language of Aztec
people. Chia plant, which name in latin is Salvia hispanica L., belongs to the
Lamiaceae family, which makes them a part of the mint family. It is
approximately a meter tall, with opposite, petiolate, and serrated leaves that are 4
to 8 cm long and 3 to 5 cm wide. The plant grows mainly in mountainous areas
and has little tolerance to abiotic phenomena, such as freezing and sunless

10
locations, however it is semi-tolerant to acid soils and drought. It is an annual herb
and usually blooms suring the summer months (Munoz et al., 2013).
Seeds of chia plant are small (1.87± 0.1 mm length, 1.21 ± 0.08 mm width
and 0.88 ± 0.04 mm thickness) with an oval flattened shape and ranged in colour
from dark coffee to beige with small darker spots (Ixtaina, et al., 2008). Its seeds
was once used by the Aztecs as a food, and acquired importance as a staple crop
in central Mexico between 1500 and 900 BC. At the time, only corn and beans
surpassed chia in term of importance (Munoz et al., 2013).
Figure 2.1 Salvia hispanica L.
Source: Munoz et al. (2013)
2.4.1 Nutritional Value of Chia Seed
Chia seed has been a subject of interest for food scientists and industries
due to the components found inside. It is rich in protein, fat, and dietary fiber
(Ayerza, 1995; Ayerza and Coates, 2005). Additionally, other components such as
vitamins, minerals, and antioxidant are also present in chia seed (Munoz et al.,
2013). Table 2.1 shows the composition of chia seed.

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Table 2.1 Composition of chia seed per 100 g
Component Content
Energy (Kcal.) 486
Proteins (g) 9 - 23
Total fat (g) 25 - 35
Saturated fatty acid (g) 3.1 - 3.4
Monounsaturated fatty acid (g) 2.309
Polyunsaturated fatty acid (g) 23.6 - 27.75
Trans fatty acid (g) 0.14
Omega-3 fatty acid (g) 17.83 g
Carbohydrate (g) 42.12
Total dietary fiber (g) 18 - 41
Vitamin C (mg) 1.6
Niacin (mg) 8.83
Thiamin (mg) 0.62
Riboflavin (mg) 0.17
Calcium (mg) 631
Potassium(mg) 407
Phosphorus (mg) 860
Iron (mg) 7.72
Source: Munoz et al. (2013); Reyes-Caudillo et al. (2007); Ayerza and Coates (1996).
Dietary fiber is found in large amount in chia seeds, which is about 18 - 41
g/ 100 g seeds (Reyes-Caudillo et al., 2007). The recommended daily intake
(RDI) of dietary fiber for adult is generally in the range of 20 to 35 g/ day. This
means that consumption of about 100 g of chia seeds can fulfill the RDI of dietary
fiber in one day (Munoz et al., 2013).
One of the most important nutritional component found in chia seed is the
fat or lipid. Its content is approximately between 25 - 35 g/ 100 g of seeds (Ayerza
and Coates, 2011; Ixtaina et al., 2011). Total polyunsaturated fatty acids (PUFA)
found in the seed is more than 80.5 % of the total fats. However, the main
constituent of the fat content in chia seed is ω-3α-linolenic fatty acid, which
consists 56.9 - 64.8 % of the total fat, the highest compared to any known plant
source (Munoz et al., 2013).

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2.4.2 Utilization of Chia Seedin Food
Chia seed utilization as a food or ingredient of food began only recently. It
was approved as a Novel Food by the European Parliament and Council of Europe
in 2009. Some of the most important applications of the seeds include its use as a
nutritional supplement and as an ingredient in cereal bars, biscuits, pasta, bread,
snacks and yogurt. Also, due to abundance of essential fatty acids in the seed, it
can be used as an oil. Another application of chia seed is using its mucilage. This
component is composed mainly of polysaccharides in form of soluble fiber. A
recent study shows that the mucilage can be used as the new source of
polysaccharides, with the potential to generate different polymer blends to
produce films and coatings with improved properties. (Munoz et al., 2013).
Pizarro et al. (2013) studied the incorporation of whole chia flour (WCF)
in pound cake. In the research, different contents of WCF (0 - 30 g/ 100 g flour
mixture) and hydrogenated vegetable fat (HVF) (12 - 20 g/ 100 g flour mixture)
were added to the cake mix based on a 22 central composite rotational design
(CCRD). The result showed that WCF addition decreased the specific volume and
colour parameters of the cakes, however, variation of WCF and HVF content
contributed to maintenance of the moisture during storage. The best formulation
was found in cake containing up to 15 g WCF/100 g flour mixture and from 16 to
20 g HVF/100 g flour mixture. The cake was found to present higher protein,
lipid, and ash content than the control cake. The omega-3 fatty acid content also
increases considerably. The sensory test of the cake shown that it has good
sensory acceptance and a greater purchasing intention.

13
Coelho and Salas-Mellado (2014) researched the effect of substituting chia
flour or seeds for wheat flour on the quality of bread. In the research, the HVF
content was reduced and chia seeds or flours were added to the formulations of
wheat flour based on a 22 CCRD. The best formulation was found on bread with
7.8 g chia flour/ 100 g flour mixture and 0.9 g HVF/ 100 g flour mixture and
bread with 11.0 g chia seeds/ 100 g flour mixture and 1.0 g HVF/ 100 g flour
mixture, which results in a reduction of 27% and 24% level of saturated fat
respectively, compared to the control bread. The ratio of PUFA and saturated fats
(PUFA:SAT) was increased to 3.1 and 3.9 respectively, compared to the control
bread (1.01). Dietary fiber content was increased to 2.0% and 5.7%, while the ω-
3α-linolenic fatty acid was increased to 1.21% and 1.85% respectively, a
significant increase compared to the control bread, which has a dietary fiber
content of 0.3% and ω-3α-linolenic fatty acid content of 0.03%. Based on the
sensory evaluation test, bread formulated with chia seeds or flour obtained high
level of acceptability test and purchase plans, with chia flour bread obtaining
higher index of purchase intent than the chia seed bread.
2.5 Omega-3 Fatty Acid
Omega-3 fatty acid is a type of polyunsaturated fatty acid. The term
omega-3 is in reference to the carbon molecule adjacent to a double bond
numbered from the methyl end of the molecule. An example of omega-3 fatty acid
is docosahexanoic fatty acid (DHA). It is a molecule with a backbone of 22
carbon chains and contains 6 double bonds, with the first double bond located on
carbon 3 from the methyl end. The other two commonly found omega-3 fatty acid

14
in food are eicosapentaenoic acid (EPA) and alpha-linolenic acid (ALA)
(McManus et al., 2011).
Among the three commonly found omega-3 fatty acids in food, ALA is
the only one not naturally produced by human body, and so it is considered to be
an essential fatty acid. However, enzymes in human body can convert ALA into
EPA and DHA, which have long been associated with health promoting effects
(Larsen et al., 2011). According to McManus et al. (2011), consumption of
omega-3 fatty acids have been related to positive health outcomes, notably in the
areas of infant development, cardiovascular disease, platelet aggregation,
hypertension, hyperlipidemia, cancer, dementia, Alzheimer’s disease, depression
and inflammation.
According to Simopoulos and Cleland (2003), maintaining an omega-
6/omega-3 ratio of 4:1 or less is recommended. High ratio of omega-6/omega-3 is
detrimental to health and can lead to the development of chronic disease.
Improving the intake of omega-3 fatty acid is essential for brain function and
management of cardiovascular disease, arthritis, and cancer. Further research by
Gow and Hibbeln (2014) also shows that omega-3 and omega-6 intake needs to be
balanced for optimal physical and mental health, and excessive intake of one type
of fatty acid may inhibit the conversion of the other. Furthermore,
supplementation of omega-3 towards children in the experiment shows some
improvement in terms of learning capacity and behavior, especially in children
who are underachieving, have ADHD-like symptoms, and/or have severe
misconduct.

15
2.6 Dietary Fiber
According to Brown (2011), dietary fiber is defined as the undigested
portion of carbohydrates remaining in a food sample after exposure to digestive
enzymes. Dietary fibers are usually found in plant foods and includes
polysaccharides and lignin. More recently, however, the definition has been
expanded to include oligosaccharides, such as inulin and resistant starches.
Dietary fibers are usually classified based on its solubility, which are soluble and
insoluble fiber. Insoluble fiber cannot be dissolved in water, and usually acts as a
sponge in the intestine by soaking up water. Soluble fibers readily dissolves in
water, and may benefit health by lowering high blood cholesterol levels and
reducing high blood glucose. Example of soluble fibers are beta-glucans, pectins,
gums, and some hemicelluloses (Anderson et al., 2009). Foods containing soluble
fibers include dried beans, peas, lentils, oats, rice bran, barley, and oranges.
Insoluble fibers are found mostly in whole wheat (wheat bran) and rye products,
along with banana. Examples of insoluble fibers are cellulose, lignin, and some
hemicelluloses (Anderson et al., 2009; Brown, 2011).
High intake of dietary fibers have been associated with a lot of health
benefits. Its main effect is to regulate intestinal degradation and absorption of
nutrients as well as their transit along the gut (Taghipoor, et al., 2014). Lower
prevalence of coronary heart disease, stroke, peripheral vascular disease, diabetes,
obesity, and certain gastrointestinal diseases have been proven by consuming high
intake of dietary fiber (Anderson et al., 2009). One of the reason is because fibers
are found to be able to lower blood pressure and serum cholesterol levels.
Improved glycemia and insulin sensitivity is also caused by high intake of dietary

16
fiber. Dietary fibers are also found to be able to significantly enhances weight loss
of obese individuals. Recent researches shows that intake of inulin and certain
soluble fibers might enhances immune function in humans, although further
research need to be done to prove it (Anderson et al., 2009).
2.7 Response Surface Methodology
Response Surface Methodology (RSM) is a collection of mathematical and
statistical techniques that are useful for the modeling and analysis of problems, in
which a response of interest is influenced by several variables and the objective is
to optimize this response (Montgomery, 2001). This method is able to process
responses based on precise maps using mathematical models to achieve one spot
that meet all of the required target. RSM is often used by industries to optimize a
certain factor in the process to achieve the best end product with minimal loss
(Montgomery, 2001).
2.7.1 Central Composite Design
Central Composite Design contains an imbedded factorial or fractional
factorial design with center points that is added with a group of “star points”
which allow estimation of a curvature (NIST, 2012). The distance from the central
to the factorial point is defined as +1, while the distance from the central point to
the star point is |α|>1. The value of α depends on the properties based on the
design and the numbers of factors involved. The star points represents the extreme
value for each of the factor involved in the design. The number of star points
always contains twice as there are factors in the design. There are three types of
central composite design, which are circumscribed, inscribed and face centered.

17
Circumscribed designs are the original form of central composite design. This
design has circular, spherical or hyperspherical symmetry and requires 5 levels for
each factor. Inscribed type is usually used whereby the limits for each factor are
truly limits. It is basically a scaled down circumscribed design with each factor
level of the design divided by α to generate the inscribed design. Face centered
type’s star points is at the center of each face of the factorial space, so α = + 1
(NIST, 2012).

18
CHAPTER III
RESEARCH METHODOLOGY
3.1 Materials and Equipments
Materials used in this experiment include chia (Salvia hispanica L.) seeds
obtained from SuperFood Indonesia in Jakarta, wheat flour with brand “Kunci
Biru”, margarine/hydrogenated vegetable fat (HVF) with brand “Blue Band”,
sugar, egg yolk, baking powder, and vanilla powder. For analysis, materials which
were used includes H2O2 ex.Merck, H2SO4 ex.Merck, K2SO4 ex.Merck, NaOH
ex.Merck, HCl ex.Merck, mixed indicator (100 mL of 0.1% methyl red with 200
mL of 0.2% bromocresol green) ex.Merck, Boric acid ex.Merck, Hexane PA
ex.Merck, Na3PO4 ex.Sigma-Aldrich, termamyl enzyme ex.Merck, pepsin enzyme
ex.Merck, pancreatin ex.Merck, ethanol ex.Merck, and acetone ex.Merck.
Equipments used in this experiment were dry blender, sifter, blender, oven,
mixer, stove, cookie cutter, balance “Mettler Toledo”, and pan. For the analysis,
equipments used were oven “Memmert”, kjeldahl system “VELP Scientifica UDK
127”, kjeldahl flask, desiccator “Duran”, muffle furnace “Thermoline 48000”,
tongs, texture analyzer “VELP Scientifica UDK 127”, evaporating dish, filter
paper, erlenmeyer, soxhlet equipment, and burette.
3.2 ResearchProcedure
3.2.1 Preliminary Research
3.2.1.1 Nutritional Content Analysis of Chia Flour

19
Chia flour was analysed for its nutritional content by using proximate
analysis. The seeds was initially grounded using dry blender before sieving
through 20-mesh screen. Proximate analysis was used to determine the
carbohydrate, protein, fat, moisture, ash content, dietary fiber and omega-3 fatty
acid.
3.2.1.2 Determination of Mixing Method
This step was done to determine the method which gives the best texture
for the cookies. Two types of mixing method were used to make the cookies,
which are one stage mixing and two stages mixing. Basic formula shown in Table
3.1 was used for preliminary research.
Table 3.1 Basic formulation for cookies
Ingredients Amount (g)
Wheat flour 100
Chia flour 15
Margarine/HVF 55
Sugar 70
Egg 40
Vanilla powder 0.2
Baking powder 0.2
Source: Novianty (2015)
In one stage mixing, all ingredients were weighed, placed in the mixer, and
then mixed at low speed until the mixture becomes uniform. For two stages
mixing, all the ingredients were weighed and then placed separately. First, the fat,
sugar, and salt were put in the mixer and then mixed at low speed until the texture
is light and fluffy. Afterwards, eggs and other liquid ingredients were added and
blended at low speed. Then, the flour and leavening agent were added and mixed
until it becomes homogen. Sensory analysis and texture analyzer was used to
determine the best mixing method (Bassinello et al., 2011).

20
3.2.2 Main Research
In the main research, the first activity done was making cookies with
different formulation of chia flour and hydrogenated vegetable fat (HVF), which
was determined according to a 22 Central Composite Rotational Design. Before
making the cookies, all chia seeds were grinded using dry blender and sieved
through a 20-mesh screen and packed in plastic containers. Afterwards, all
ingredients used in the formulation, which are wheat flour, chia flour, HVF, sugar,
egg, vanilla powder, and baking powder were weighed according to the
formulations, shown in Table 3.3. Next, all the ingredientes were mixed together
using the proper mixing method, which was determined in the preliminary
research. Using a cookie cutter, the dough was shaped and then placed into
margarine-smeared pan. After that, the dough was baked at 145 oC for
approximately 15 - 20 minutes. The cookies were cooled down after the baking
process is finished. Figure 3.1 shows the flowchart of cookie production.
The second activity was evaluating the physico-chemical characteristics of
the cookie produced, which includes the spread of the cookies, texture, color, and
moisture content. The third activity was sensory test to evaluate the preference of
panelists towards the cookie produced using hedonic test. After the result from the
physico-chemical analysis and sensory evaluation comes out, Response Surface
Method (RSM) was used to determine the recommended best composition.
Verification test was done afterwards towards the recommended composition.
After the optimum or best cookie formulation is determined, proximate analysis
was done to analyse the difference of the components inside the cookie between
cookies made with chia flour and the control cookie. The analysis includes

21
carbohydrate content, protein content, fat content, ash content, fiber content, and
omega-3 content. The last activity was comparing the physical characteristics and
the result of the proximate analysis between the experimental cookies and control
cookies.
Grinding chia seeds into flour using dry blender and sieved through 20-mesh screen
Ingredient are weighted according to formulation (refer to Table 3.3)
Chia and wheat flour are mixed with HVF, sugar, egg, vanilla powder, and baking powder with the
proper method determined from preliminary research
The dough is shaped using cookie cutter, and then placed into the margarine-smeared pan
The dough is baked at 150oC for 15 minutes
Let the cookies cool
Figure 3.1 Flowchart of the cookies making
Source: Novianty (2015), with modification
3.2.2.1 Determination of Chia Cookie Formulation
Detemination of formula used to produce chia cookies was conducted
using RSM with Design Expert 7.0®. This method was used to help the
optimization of composition of component used in the cookies, by recommending
the best formulation to be used based on responses to each formulation. There are
two factors used in this research, which are chia flour and HVF. Table 3.2 shows
the upper and lower level of chia flour and HVF used, and the cookies formulation
for optimization can be seen in Table 3.3. Table 3.4 shows the goal and
importance of each parameter. Texture parameter was set to the highest level,
which is 5, as it is the main objective of this research. Sensory evaluation result
was done to determine which cookie formulation have the highest acceptability in
texture parameter. Afterwards, physico-chemical analysis was done to determine
the effect of different compositions of chia flour and HVF towards the texture

22
characteristics of cookie. The RSM recommended the optimal composition to
produce chia cookies.
Table 3.2 Concentration composition of cookies formula
Factor -α Level (g) -1 Level (g) 0 +1 Level (g) +α Level (g)
Chia Flour 0.00 4.39 15 25.61 30
HVF 33.00 36.22 44.00 51.78 55
Table 3.3 Formulation Combination Model
No Chia Flour (g) HVF (g)
1 4.39 36.22
2 15.00 44.00
3 4.39 51.78
4 25.61 51.78
5 25.61 36.22
6 15.00 44.00
7 15.00 44.00
8 15.00 55.00
9 30.00 44.00
10 15.00 44.00
11 15.00 33.00
12 15.00 44.00
13 0.00 44.00
14 15.00 44.00
Table 3.4 Optimization cookies formula goal and importance
Factor Goal Importance
Organoleptic Texture Maximize 5
Physico-chemical Hardness Is in range 3
characteristics Fracturability Is in range 3
Cookies spread Is in range 3
Moisture content Is in range 3
3.2.2.2 Verification Test
This test was done to verify the formula given by the RSM. It was done by
producing cookies based on the recommended formulation and then conduct
physico-chemical analysis and sensory evaluation towards the cookies. The result
of the tests was compared with the prediction value of RSM.

23
3.2.2.3 Product Comparison
To observe the effects given by the addition of chia flour, comparison was
done by comparing the experimental cookies with control cookies. The nutritional
content, physico-chemical properties, and sensory evaluation result is compared
with control cookies.
3.3 Experimental Design
3.3.1 Preliminary Research
The treatment done in the preliminary research was to compare the effects
of different mixing method towards the physical characteristic of the cookies
produced. The data was analysed using binomial test with two factors, which was
one-stage and two-stage mixing method. The hypothesis are as follows:
H0 = There is no effect of different mixing method towards panelist preference.
H1 = There is an effect of different mixing method toward panelist preference.
3.3.2 Main Research
3.3.2.1 Chia Cookies Formula Determination
The experimental design for this research was done to determine the best
formulation of cookies using the optimal concentration of chia flour and HVF
using RSM method. The Design Expert processed the result of the analysis and
recommend the best formulation. The statistical model of this research is written
below:
Y = β1X1 + β2X2
Y = Response function
β = Linear coefficient
X1 = Chia flour concentration
X2 = HVF concentration

24
3.3.2.2 Product Comparison
Components and physical properties of experiment cookies and control
cookies were compared using independent sample t-test. Dependent sample t-test
was used when comparing the sensory parameters between experiment and
control cookies. The statistical model is as follows:
Yij = μ + Ai + εij
Yij = Observation value from experiment cookies at level 1 and repetition j.
μ = Actual mean value
Ai = Effect of formulation at level i
εij = Error factor
H0 = There is no difference between experiment cookies and control cookies
H1 = There is a difference between experiment cookies and control cookies
3.4 Analysis Procedure
3.4.1 Physico-chemical Analysis
3.4.1.1 Cookies texture (Bourne, 2002)
Texture analyzer was used to determine the cookie texture. Hardness value
was considered as the maximum force obtain in the curve and fracturability as the
linear distance towards the maximum point. The studies was conducted using a 2
mm probe at a crosshead speed of 3 mms-1. Parameters measured were hardness
and fracturability.
3.4.1.2 Cookies spread (Sharif et al., 2009)
Spreading of cookie was observed manually using a ruler. The height and
diameter of the cookies before and after baking was observed as well. The height
was observed by placing six cookies horizontally (from edge to edge), and has its
height measured while the cookies are rotated 90o. Thickness was measured by
placing six cookies to one another.

25
Spread factor =
Weight
Thickness
x Correction Factor x 10
3.4.1.3 Proximate Analysis
3.4.1.3.1 Moisture Content (AOAC, 2005)
Analysis of moisture content was done using oven method. First, 5 grams
of sample was transferred to the constant evaporating dish and placed in the oven
at 105 oC for 6 hours. After drying, the sample was moved into a desiccator to
cool down. Weighing process was done afterwards, and finished until the constant
weight is achieved.
Moisture Content (Wet Basis) =
W1−W2
W1
x 100
Where: W1 = weight (g) before drying
W2 = weight (g) after drying
3.4.1.3.2 Ash Content (AOAC, 2005)
Determination of ash content of cookies was done using dry ashing
method. Approximately 3 g of sample was weighed and put in the constant
crucible, which was digested until no smoke is formed. The crucible was then
placed in a muffle furnace, which afterwards was ignited at 550 oC until light gray
ash was obtained. The obtained ashes was then cooled in desiccator and weighed.
% Ash content (dry basis) =
x−y
z
× 100%
where: x = weight after ashing
y = weight of crucible
z = original sample weight
3.4.1.3.3 Protein Content (AOAC, 2005)

26
Kjeldahl method was used to analyze the protein content. Fat free sample
weighing around 2 g was wrapped with a filter paper and placed in digestion flask.
The flask was then added with 7 g of K2SO4 and 0.05 g of Selenium. It was then
followed by the addition of 10 mL H2SO4 and 10 mL H2O2. The sample was
destructed until clear solution was obtained. Afterwards, the solution was cooled
down with the addition of 50 mL NaOH. The flask was then connected to a
distilling bulb on a condenser, in which the tip was immersed in a solution of
boric acid mixed with 3 drops of indicator. The distillation process was done for
about 5 minutes. Any excess acid in the distillate was then titrared using HCl
solution until pink color is achieved.
% Protein =
((𝐴−𝐵) 𝑥 𝑁))𝑥 14.007 𝑥 6.25
W
x 100%
Where: A = Volume (ml) of titration
B = Volume (ml) of blank
N = N HCl
W = weight (mg) of sample
14.007 = atomic weight of nitrogen
6.25 = protein conversion factor
3.4.1.3.4 Fat Content (AOAC, 2000)
Fat content analysis was done using soxhlet method. Approximately 5 g of
sample was wrapped in filter paper and put into extraction thimble. Hexane was
then added into the boiling flask. Afterwards, boiling flask, soxhlet flask, and
condenser was assembled. Fat was then extracted in a soxhlet extractor for 6
hours. Then the boiling flask with extracted fat was dried in oven at 105 oC until
constant. After drying, the sample was cooled down in the desiccator, and then
weighed.
% Fat content (dry basis) = weight of fat extracted (g) × 100%
Weight of sample (g)

27
3.4.1.3.5 Carbohydrate Content (AOAC, 2005)
Determination of carbohydrate content was done using difference method.
Carbohydrate content (%) = 100% - (% moisture + % ash + % protein + % fat)
3.4.1.4 Dietary Fiber Content
Approximately 1 g of dry fat free sample was placed in Erlenmeyer, and
0.1 M sodium phosphate is added. Afterwards, 0.1 ml of termamyl enzyme was
added into the mixture, and then covered with aluminium foil. The mixture was
then incubated in a water bath at 100 oC for 15 minutes. The mixture was then
cooled down, and afterwards 20 ml aquadest and 1 M HCl was added. The pH of
this mixture should be 1.5. Afterwards, 100 mg of pepsin enzyme was added, and
then the erlenmeyer flask was covered again and incubated in water bath at 40 oC
for 60 minutes. HCl addition was done to adjust the pH to 4.5. The mixture was
then filtered with a dry crucible which have been weighed and contains 0.5 g of
dry celite. The mixture was then washed twice with 10 ml of aquadest. The filtrate
was then used to determine the soluble fiber, while the precipitates were used to
determine the insoluble fibers. The precipitate was washed with 10 ml of 95%
ethanol and 10 ml acetone twice. Afterwards, the precipitate was dried in 105 oC
until it reaches constant weight. Then, the precipitate was ashed in furnace at 550
oC for 5 hours.
% Insoluble fiber = (A – B – C)/W x 100%
A = weight after dried (g)
B = weight after ashed (g)
C = weight of fat free blank (g)
W = sample weight (g)

28
The soluble fiber was determined by adding water into the resulting filtrate
until the volume reaches 100 mL. Afterwards, 400 ml of 95% ethanol was added
and the mixture was cooled for 1 hour. The mixture was then filtered with dry
crucible which have been weighed and contains 0.5 g of celite. The resulting
filtrate was washed twice with 10 ml of 78% ethanol, 10 ml 95% ethanol, and 10
ml acetone. Afterwards it was dried in 105 oC until constant weight was reached.
The filtrate was then ashed in furnace at 550 oC for 5 hours and cooled down in
desiccator before weighing.
% Soluble fiber = (A – B – C)/W x 100%
A = weight after dried (g)
B = weight after ashed (g)
C = weight of fat free blank (g)
W = sample weight (g)
Total dietary fiber = insoluble fiber + soluble fiber
3.4.1.5 Omega-3 Fatty Acid Content
Omega-3 analysis was done using gas chromatography. Initially, the
samples were dried and has its fat content extracted, using fat extraction method
from AOAC (2000). Afterwards, the fatty acid methyl esters (FAMEs) found in
the fat were obtained and the compositions determined via gas chromatography
with a flame ionisation detector (FID). Omega-3 fatty acid was determined and
has its content calculated, with the result expressed as g per 100 g of fat (AOCS,
2005).
3.4.2 Sensory Evaluation
Hedonic test was used in the sensory evaluation. Texture parameter is used
in this test. A total of 75 panelists was used. In the test, 1 indicates the least
preferred cookie, while 7 indicates the most preferred cookie. Panelists were

29
asked to determine the score by eating each of the cookie sample. Mineral water
was provided and panelists must take a sip of water in between tasting each of the
samples. Afterwards panelists have to write the score in the form (Meilgaard et
al., 2007).

30
CHAPTER IV
RESULT AND DISCUSSION
4.1 Overview
The cookies made were pressed cookies, with the addition of chia seed
flour. Proximate analysis of the chia seeds and determination of the best mixing
method were done first before the main experiment was commenced. The main
experiment was done to determine the optimal cookie formulation, the verification
of the formula, and characterization of the product.
4.2 Nutritional Content of Chia Flour
Chia flour is the main component in the making of the cookies. The
nutritional content of chia flour can be seen in Table 4.1. The identification result
is written in Appendix A.
Table 4.1 Nutritional Content of Chia Flour
Nutrition Chia Seed
Protein (%) 17.24 ± 0.10
Fat (%) 30.97 ± 0.25
 Omega-3 ( %) 19.14
Ash (%) 1.66 ± 0.13
Carbohydrate (%) 41.88 ± 0.35
Total Dietary Fiber (%) 42.60
Moisture (%) 8.25 ± 0.38
The nutritional value of chia flour closely matches the value from USDA,
in which it is written as follows: 16.54 g protein/100 g, 30.74 g fat/100 g, 17.83 g
omega-3 fatty acids/100 g, and 42.12 g carbohydrate/100 g. However the total

31
dietary fiber content of chia flour was significantly higher than the value from
USDA, which is written as 34.4 g dietary fiber / 100 g (Munoz et al., (2013).
4.3 Determination of Mixing Method
The mixing method of the cookies was determined based on the sensory
evaluation of the cookie’s texture. As fibers are found in large amount in chia
seeds, and it can affect the texture of the cookie produced, hence texture is the
parameter analyzed. Based on the result of the sensory evaluation, the best mixing
method is used. Two mixing method used in this experiment are one-stage and
two-stage mixing. Paired comparison and paired preference test were used in the
determination of mixing method. In paired comparison test, panelist were asked to
determine which cookie is more hard to bite, while in paired preference test, the
preferred cookie is chosen. Appendix B shows the questionnaire of both tests. The
result of the test is as shown in Figure 4.1 and 4.2.
Figure 4.1 Result of Paired Comparison Test
Note : One-stage = 72 panelists; Two-stage = 3 panelists
96%
4%
One-stage
Two-stage

32
Figure 4.2 Result of Paired Preference Test
Note : One-stage = 12 panelists; Two-stage = 63 panelists
The sensory evaluation data of both tests are found in Appendix C, while
the statistical analysis are found in Appendix D. The result from both the paired
comparison and paired preference test shows that there are significant differences
between cookies made with one-stage and two-stage mixing. In the paired
comparison test, cookie made with one-stage mixing is significantly harder than
cookie made with two-stage mixing. The paired preference test shows that cookie
made with two-stage mixing is more preferable than cookie made with one-stage
mixing. The result is backed by result from texture profile analysis, which can be
seen in Appendix E. Table 4.2 shows the average of hardness value of cookie
made with one-stage and two-stage mixing respectively. According to Hui (2006),
mixing cookie ingredients in two stages can minimize the development of gluten
in the flour, which is desirable in cookie dough. By mixing the cookie ingredient
in two stages, the fat will be more evenly dispersed around the flour particles, and
therefore rendering them less available to water and preventing gluten
development. High protein content, which causes gluten development in cookie
16%
84%
One-stage
Two-stage

33
dough is associated with harder cookies (Gaines, 1992). In one-stage mixing, the
baker has less control over the mixing, and therefore, the gluten development as
well. However, it can be used when over-mixing is not a serious problem, for
example in the making of chewy cookies (Hui, 2006).
Table 4.2 Hardness Value of Cookie Made With Different Mixing Methods
Mixing Method Hardness (g)
One-stage mixing 2944,10 ± 156,54
Two-stage mixing 1864,33 ± 176,78
4.4 Formula Optimization of Cookies
This step was done to find the formula to made cookies from chia seeds
with the best texture based on the sensory evaluation. The sensory evaluation
questionnaire can be seen on Appendix F. Parameter evaluated is texture. The test
was done in the span of two weeks, in which the panelists must evaluate the
sensory attributes of 3 and 4 samples each week, with a total of 14 samples, which
was done to avoid sensory fatigue. According to Stone and Sidel (2004), sensory
fatigue can happen if panelists are asked to evaluate too many samples, and
consequently, the evaluation result becomes inaccurate. The result of the sensory
evaluation can be seen on Appendix G. Texture analysis, moisture content, and
spread rate were used to obtain objective data regarding the parameters of the
cookies.
The data obtained from the experiment were analyzed using Response
Surface Methodology (RSM). RSM is a mathematical and statistical method to
optimize responses from several variables (Montgomery, 2001). In this case, the
responses are all the parameters which was measured and the variables are the
chia flour and hydrogenated vegetable fat (HVF). Through RSM, the best

34
combination of chia flour and HVF to obtain cookies with the optimal texture can
be found. The analysis result from RSM shows the appropriate statistical model
for each responses. Table 4.3 shows the appropriate statistical model of each
responses.
Table 4.3 Statistical Model for Each Responses
Response Model P-value model P-value Lack of fit
Texture Linear 0.0011 0.4712
Hardness Linear 0.0003 0.4558
Fracturability Quadratic 0.0003 0.2622
Spread ratio Quadratic 0.0001 0.0404
Moisture content Linear 0.0026 0.7306
In RSM, the P-value of the statistical model must be significant to show
that it is the most appropriate model (<0.05). While for the P-value in lack of fit
must be not significant to indicate that the chosen model is appropriate (>0.05)
(NIST, 2012). As shown in Table 4.3, all the responses has P-value less than 0.05,
which means all the selected models are significant, indicating that there are
significant differences found between the different compositions of chia flour and
HVF. However, in lack of fit, the P-value of spread ratio was found to be
significant, which means that the selected model does not accurately fit the data,
however, as the P-value model is significant, it means that although the different
composition of chia flour and HVF affected the spread ratio significantly, the
selected model does not represent the data accurately.
4.4.1 Optimization of Sensory Evaluation Result
Sensory evaluation was done to evaluate the preference of panelists in
terms if its texture. The texture of cookies is the most important parameter in
cookies. What differentiates cookies with the other bakery product is its crispiness.

35
The interaction between fat and flour used is what made the characteristic
crispiness of the cookies. The resulting interaction between chia flour, wheat flour,
and HVF in this experiment were measured in terms of desirability in the sensory
evaluation test. Table 4.3 shows the P-value for model and lack of fit in fit
summary test, and linear model was found to be the most appropriate model to
represent the data. Figure 4.3 shows the 3-D graph for texture response, while
Figure 4.4 shows the contour graph. Appendix H shows the statistical analysis of
texture response. According to the analysis, chia flour and HVF significantly
affects the texture response. Shown below is the final equation for texture
response:
Y= 2.54849 - 0.033123 A + 0.066099 B
Note:
Y = Hedonic score for texture
A = Chia flour
B = HVF
Figure 4.3 3-D Graph for Texture Response Based on Hedonic Test

36
Figure 4.4 Contour Graph for Texture Response Based on Hedonic Test
The result of the sensory evaluation test shows that panelists prefer
cookies made with less chia flour and high amount of HVF. From the graph, it can
be concluded that high concentration of HVF can offset the undesirability of high
amount of chia flour. The result is similar to experiments done by Pizarro et al.
(2013) and Steffolani et al. (2015), whereby pound cake and bread added with
chia flour decreases its texture desirability respectively.
4.4.2 Optimization of Physical Analysis Result
4.4.2.1 Hardness
Hardness is considered to be the force needed to bite through the cookie in
a single bite (Bourne, 2002). Fit summary test from RSM in Table 4.2 shows that
linear model is suitable to represent the hardness response. The graph for hardness
response can be seen in Figure 4.5 and 4.6. Appendix I and Appendix J shows
result of hardness analysis and the statistical analysis of the hardness response
respectively. The statistical analysis result shows that both chia flour and HVF

37
significantly affects the hardness response. The equation for hardness response is
as follows:
Y= 5754.73699 + 12.08214 A - 62.86553 B
Note:
Y = Hardness value
A = Chia Flour
B = HVF
Figure 4.5 3-D Graph for Hardness Value
Figure 4.6 Contour Graph for Hardness Value

38
Based on the result shown in the graph and equation, it can be seen that
chia flour significantly increases the hardness of cookie, while HVF decreases it.
According to Pizarro et al. (2013), chia seed contains high amount of dietary
fibers, which in turn disturbs the fat and air distribution in the dough by exerting
physical impairment towards the dough.. Fat distribution is especially important in
cookie, as fat impedes gluten formation by breaking the long gluten strands in the
dough and help stabilize air cells. According to Gaines (1992), gluten formation is
associated with hard cookies, as the gluten structure . With the fat distribution
interrupted, gluten formation occurs, and consequently the cookies become harder
due to gluten’s tough and elastic nature. The result, compared with the sensory
evaluation result of texture parameter shows that cookies with hard texture is
undesirable according to panelists. Therefore, it can be concluded that the addition
of chia flour increase the hardness of cookie, and HVF can help decrease it.
4.4.2.2 Fracturability
According to Bourne (2002), fracturability is the force needed to shatter a
cookie in a single bite. Based on fit summary test, quadratic model fits the
fracturability response data. Figure 4.7 and 4.8 shows the graph for fracturability
response. Red color indicates high value and low value is indicated with blue
color. The result of fracturability analysis is shown in Appendix I. Appendix K
shows the statistical analysis of fracturability response. According to the analysis,
all the factors are significant, except for the quadratic HVF (B2) factor. This is
because the p-value is above 0.05, meaning that the factor does not significantly
affect the fracturability response and as a result, it is not put on the final equation.
Below is the equation for the fracturability response:

39
Y= 11.70780 + 0.087422 A - 0.021202 B - 3.94545 x 10-3 AB + 1.61889 x 10-3 A2
Note:
Y = Fracturability value
A = Chia Flour
B = HVF
Figure 4.7 3-D Graph for Fracturability Value
Figure 4.8 Contour Graph for Fracturability Value
Based on the interaction term and the graph, it can be seen that the
addition of HVF significantly increases the fracturability of cookie in low amount

40
of chia flour, but the effect is not significant if the chia flour is added in large
amount. Addition of HVF higher than used in this research might be needed to
maintain high fracturability value. Chia flour have a very small positive quadratic
effect towards the fracturability of cookie, whereby in small and high amount, the
value actually increases. Similar to the texture response, the change in
fracturability value is also affected due to the large amount of dietary fiber found
in the chia flour (Pizarro et al., 2013). As more chia flour is added, so does the
dietary fibers, therefore causing the fat to be unevenly distributed. This is because
fiber exerts physical impairment toward the dough, by displacing the fat
distribution around the dough. According to Jacob and Leelavathi (2006), fat
helps increases the crispiness of cookie, which is associated with high
fracturability. This is due to fat’s ability to break long gluten strains in the dough,
which consequently causes the dough to soft and less viscous. The resulting
dough after baking will be soft and crispy. Due to uneven distribution of fat, these
long gluten strains were not broken and the cookie becomes harder to fracture,
due to gluten’s tough and elastic nature. This is supported by research done by
Novianty (2015), whereby the incorporation of ingredient with high amount of
dietary fiber decreases the fracturability of cookies. The result, compared with the
result of sensory evaluation in texture parameter, shows that low fracturability
value of cookie is undesirable according to panelists. Therefore it can be
concluded that chia flour significantly decreases the fracturability of cookie, while
HVF can increase it in low amount of chia flour.

41
4.4.2.2 Spread Ratio
Spread ratio is defined as the ratio between the diameter of cookies and its
thickness. The value is directly proportional of its diameter and inversely with its
thickness. Fit summary test from RSM shown in Table 4.3 shows that the
quadratic model fits the data for spread ratio. However, the P-value for lack of fit
was shown to be significant. The possible explanation for this is the high standard
deviation of the data. The result and statistical analysis for spread ratio response is
shown in Appendix L and Appendix M respectively. Result of the statistical
analysis shows that the quadratic HVF factor (B2) is not significant (p-value >
0.05), therefore it is not included in the final equation. Figure 4.9 shows the 3-D
graph for spread ratio response, with blue color indicating low spread ratio and
red color indicating the otherwise. The contour graph is shown in Figure 4.10.
Below is shown the equation for spread ratio response:
Y= -0.63690 + 0.12724 A + 1.37668 B - 0.023727 AB + 0.013871 A2
Note:
Y = Spread ratio value
A = Chia Flour
B = HVF
Figure 4.9 3-D Graph for Spread Ratio

42
Figure 4.10 Contour Graph for Spread Ratio
From the result of the experiment represented on the graph, it can be seen
that HVF significantly increases the spread ratio of cookie in low amount of chia
flour, however the effect is not significant if the chia flour concentration is higher.
Chia flour have very small positive quadratic effect towards the spread ratio of
cookie, whereby in small and high amount, the value actually increases.
According to experiment done by Pizarro et al. (2013), high fiber content of chia
seed lowers the specific volume of cake formulated with the addition of chia flour.
The author concluded that the cause is the high amount of fiber found in chia seed,
which interferes the distribution of fat and air around the dough. As fat were
distributed unevenly, gluten formation occurs uncontrollably, thereby causing the
dough to be tough and elastic, making it harder to spread during baking. Another
experiment done by Saeed et al. (2012) which studies the effect of sweet potato
flour on the quality of cookie also reports that high content fiber decreases the

43
width or diameter of the cookies produced. The journal mentions that high water
absorption capacity of sweet potato flour also contributes toward the decreasing
spread ratio of the final product, which decreases the amount of free water present
in the dough. The loss of free water creates viscous dough, which in turn
decreases the spread ratio in its final product. Based on manuscript written by
Munoz et al. (2013), chia seed is known to possess high water absorption capacity
as well, as the insoluble fibers has the ability to absorb high amount of water, and
as such, it further decreases the spread ratio of the final product. In this
experiment, spread ratio seems to be increased in low amount of chia flour by
increasing HVF content, as the fiber content is still relatively low, however when
the chia flour concentration is high, the fiber content becomes so high that
increasing HVF will not have any effect any longer. Increasing the amount of
HVF higher than used in present study might be beneficial to help increase spread
ratio in high amount of chia flour.
4.4.2.3 Moisture Content
According to the fit summary test, linear model fits the data for moisture
content response. Moisture content is one of the most important parameter of
cookie, as it influences the texture of the cookie formed. Low moisture content
increases the crispiness of cookie, while higher moisture content form soft cookie
(Hui, 2006). Appendix N shows the data of moisture content, while Appendix O
shows the statistical analysis for moisture content response. All factors in the
experiment significantly affects the moisture content response according to the
statistical analysis. The data for the moisture content response is represented in a

44
3-D and contour graph in Figure 4.11 and 4.12 respectively. The resulting
equation for moisture content response is as follows:
Y= 6.16682 + 0.056443 A - 0.072952 B
Note:
Y = Moisture content
A = Chia Flour
B = HVF
Figure 4.11 3-D Graph for Moisture Content
Figure 4.12 Contour Graph for Moisture Content

45
Chia flour was found to significantly increases the moisture content of
cookie. The possible explanation for this is the water absorption capability of chia
flour. According to Munoz et al. (2013), one of chia seed’s characteristic is its
ability to absorb large amount of water, due to its insoluble fiber content.
Experiment done by Novianty (2015) shows that the addition of ingredient with
high water absorption capability can increase the moisture content of cookie. The
insoluble fibers absorb free water, thereby making the dough more viscous. This
creates harder and less crispy cookies, which correlates with the result of hardness
and fracturability parameter.
4.4.3 Optimal Formula Recommendation
The optimal formula of chia flour and HVF in the cookie was determined
by Design Experiment via RSM, based on the data of the experiment. To
determine the optimal composition, goals were set for each responses and
variables, with particular focus on the texture parameter from the sensory
evaluation result, as the goal of this response is to obtain cookie with optimal
texture. To help this, the importance was set, with 5 as the highest priority and 1
as the lowest priority. The Design Expert optimized the formulation according to
goals with highest importance first, followed by the least important. Chia flour
and texture’s importance was set to 5, as the objective of this research is to obtain
cookie with highest amount of chia flour while having the best texture quality as
well. The criteria and goals for each response can be seen in Table 4.4.

46
Table 4.4 Criteria and Goals of Each Responses for Optimal Cookie Formulation
Factors Goal Lower Limit Upper Limit Importance
Chia Flour (g) Maximize 0 30 5
HVF (g) In range 33 55 3
Texture Maximize 3.95 5.84 5
Hardness (g) In range 2361.19 3930.77 3
Fracturability (mm) In range 12.028 13.637 3
Spread ratio In range 27.59 44.295 3
Moisture content (%) In range 2.65 6.01 3
Based on goals set previously, the Design Expert determined the optimal
composition of chia flour and HVF and gave the desirability value. The
desirability value is between 0 to 1, and the target is to achieve value as high as
possible, with 1 as the ideal response value (NIST, 2012). The optimal formula
recommended by the Design Expert can be seen in Table 4.5. Afterwards, the
optimal formula needs to be verified using sensory evaluation and physico-
chemical analysis and compared to the prediction value calculated by Design
Expert.
Table 4.5 The Optimal Formula of Cookie Formulated With Chia Flour
Formulation Chia Flour (g) HVF (g) Desirability
1 20.45 55.00 0.702
4.5 Product Verification
This step was done to verify the formulation recommended by the Design
Expert. In order to verify the formulation, sensory evaluation and physico-
chemical test was done. Appendix P and Appendix Q shows the questionnaire and
result of the sensory evaluation test respectively. Appendix R shows the
physicochemical analysis result. The comparison between the prediction value
and the actual result is shown in Table 4.6.

47
Table 4.6 Comparison Between Prediction Value and The Actual Result
Response Predicted Actual
95% CI
low
95% CI
high
95% PI
low
95% PI
high
Texture 5.51 5.25 5.07 5.94 4.66 6.36
Hardness 2544.11 2574.84 2242.85 2845.38 1955.12 3133.10
Fracturability 12.53 12.83 12.22 12.83 12.06 12.99
Spread Ratio 31.39 30.16 28.55 34.23 27.08 35.70
Moisture content 3.31 3.80 2.66 3.95 2.05 4.57
Based on the result in the table, it can be seen that some value are larger
than the predicted value, and some are smaller. However, as the value is still
within the 95% confidence interval (CI) and prediction interval (PI), it is still
acceptable. The 95% CI is defined as the range in which the process average is
expected to fall into 95% of the time, while 95% PI is the range in which and
individual value is expected to fall into 95% of the time (NIST, 2012).
4.6 Product Comparison
4.6.1 Sensory Evaluation Result
Hedonic test was done to determine the panelists preference between
control cookie and optimal chia cookie. The parameters observed are color, aroma,
texture, and taste. Appendix S and Appendix T shows the questionnaire and data
from the sensory evaluation. The data was further analysed using dependent
sample t-test, and is shown in Appendix U. The result of the sensory evaluation
test can be seen in Table 4.7.
Table 4.7 Result of Sensory Evaluation Test
Parameter Optimum Control
Aroma 5.56 ± 0.89 a 5.77 ± 0.92 a
Color 5.32 ± 1.22 a 5.87 ± 0.70 b
Taste 5.61 ± 0.84 a 6.09 ± 0.74 b
Texture 5.25 ± 1.24 a 6.08 ± 0.88 b
Note: Score 1: Least Desirable 7: Most Desirable

48
According to t-test analysis, there is no significant difference in terms of
aroma between the control cookie and optimum chia cookie. However, for color,
taste, and texture, the result significantly shows that panelist prefer the control
cookie compared to the optimum chia cookie. Color parameter of optimal chia
cookie was lower than control cookie due to greyish color formed in its texture,
which is similar to experiment done by Steffolani et al. (2015) regarding the
addition of chia flour into bread. The result for taste and texture also matches the
result of experiment done by Pizarro et al. (2013) about the addition of chia flour
into pound cake, which was lower compared to the control cake. Texture
parameter is lowered possibly due to higher hardness and lower fracturability of
optimum chia cookie compared to control cookie, caused by high content fiber of
chia seed which disrupts fat distribution, thereby allowing gluten to form in the
dough. The average scores for aroma, color, taste, and texture indicates that
optimum chia cookie is quite acceptable according to the panelists.
4.6.2 Physico-chemical Analysis
Parameters observed in physico-chemical analysis are hardness,
fracturability, spread ratio, and moisture content. Appendix V shows the data from
the physico-chemical analysis. Independent sample t-test analysis of the data
obtained can be seen on Appendix W. Table 4.8 shows the result of the physico-
chemical analysis.
Table 4.8 Result of Physico-chemical Analysis
Parameter Control Optimum
Hardness (g) 2017.13 ± 240.43 a 2574.84 ± 220.38 b
Fracturability (mm) 14.55 ± 0.34 a 12.83 ± 0.40 b
Spread ratio 37.38 ± 1.13 a 30.16 ± 1.12 b
Moisture Content (%) 2.31 ± 0.24 a 3.80 ± 0.45 b

49
Based on t-test analysis, all the parameters tested between the control
cookie and optimum chia cookie differ significantly. Control cookie has
significantly lower hardness and higher fracturability value compared to optimum
chia cookie. This is caused by high amount of fiber found in chia seed, which
impedes fat distribution, causing gluten to be formed which hardens the cookie
texture. Spread ratio of control cookie was also found to be significantly higher
than the optimum chia cookie, which is caused by high amount of fiber in chia
seed. The fiber impedes spread of fat, which is important in helping cookie dough
to spread as it cooks in the cooking sheet. Optimum chia cookie has significantly
higher moisture content compared to the control cookie, due to high water
absorption capacity of chia seed, which has high amount of unsoluble fiber.
4.6.3 Nutritional Composition
The nutritional value of optimum chia cookie is compared with control
cookie. This is done in order to know whether there are differences between the
nutritional value of optimum chia cookie and control cookie. The result of the
analysis is shown in Appendix X. Table 4.9 shows the data for nutritional content
of both optimum chia cookie and control cookie.
Table 4.9 Nutritional Content of Optimum Chia Cookie and Control Cookie
Nutrition Optimum Control
Protein (%) 8.75 ± 0.01 7.21 ± 0.10
Fat (%) 27.81 ± 0.05 28.10 ± 0.83
 Omega-3 ( %) 1.64 0.09
Ash (%) 1.18 ± 0.06 0.90 ± 0.01
Carbohydrate (%) 58.46 ± 0.64 61.48 ± 1.21
Total Dietary Fiber (%) 9.16 8.32
Moisture (%) 3.80 ± 0.53 2.31 ± 0.29
As seen on Table 4.9, the nutrition content of optimum chia cookie and
control cookie is only slightly different. The protein, total dietary fiber, and

50
moisture content of optimum chia cookie is slightly higher compared to control
cookie. The carbohydrate content of control cookie is slightly higher than
optimum chia cookie, while the fat content between the two cookies is very
similar. However, the most significant difference between optimum chia cookie
and control cookie is in its omega-3 fatty acid content, which was around 1.64 %
(1636.10 mg/100 g) compared to 0.09 % (87.45 mg/100 g) found in control
cookie.
According to standard set by FDA (2013), food is said to be a good source
of fiber if it contains 10 - 19 % or more of the daily value per reference amount
customarily consumed (RACC). The RACC of cookie is 30 g, and the daily value
for fiber is 25 g. The fiber content of optimum chia cookie is 9.16 %, which is
2.75 g. The amount is about 10.99 % of the daily value for fiber, meaning that it
can be claimed as a good source of fiber.
FDA (2014) allows food to be claimed high in α-linolenic acid (ALA) if it
contains ALA higher than 320 mg per RACC. Omega-3 fatty acid found in chia
seed is exclusively ALA, and so the standard can be used (Munoz et al., 2013).
The ALA content found in optimum chia cookie is 1636.10 mg/ 100 g, which
means the value per RACC is 490.83 mg. This value is significantly higher than
the minimal value, meaning that optimum chia cookie can be claimed to be a high
source of ALA.

51
CHAPTER V
CONCLUSION AND SUGGESTION
5.1 Conclusion
The best mixing method to produce chia cookie with a better texture is
two-stage mixing method. Two-stage mixing produce softer cookies than one-
stage mixing method. High amount of chia flour lowers the spread ratio of the
cookie dough. It also formed hard and less crispy cookies, and increases its
moisture content, which makes the cookie less desirable in terms of texture. HVF
is important to maintain the cookie’s softness, crispiness, and decrease its
moisture content. Spread ratio of cookie dough was also increased in high amount
of HVF. The best formulation to produce chia cookie with the optimum texture is
20.45 g of chia flour and 55.00 g of HVF respectively. The texture parameter of
the cookie produced are as follows: moisture content 3.80%, hardness value
2574.84 g, fracturability value 12.83, spread ratio 30.16. It was well liked by
panelists in sensory evaluation test, obtaining the average score of 5.25 in the
texture parameter. The experimental cookie was found to have high amount of
omega-3 fatty acids and dietary fibers.
5.2 Suggestion
Further addition of HVF might be beneficial to further optimize the
texture, however consideration needs to be taken considering the amount of
saturated fatty acid in it. Addition of reducing agent might be beneficial to
increase spread ratio of dough and reducing hardness of cookie produced.

52
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Elsevier, 2004.
Sumnu, Servet Gulum and Serpil Sahin. Food Engineering Aspects of Baking
Sweet Goods. New York: Springer, 2006.
Taghipoor, M., G. Barles, C. Georgelin, J.R. Licois, P. Lescoat. “Digestion
Modeling in the Small Intestine: Impact of Dietary Fiber”. Mathematical
Biosciences 258: 101 - 112, 2014.

A-1
Appendix A. Identification Result
Protein
Sample mL HCl Weight(g) % Nitrogen % Protein Average
1 20,84 2,1251 2,75 17,17
17,24
2 20,16 2,0394 2,77 17,31
Calculation:
% Protein =
((𝐴−𝐵) 𝑥 𝑁))𝑥 14.007 𝑥 6.25
W
x 100%
= 20,84 mL x 0,2 N x 14,007 x 6,25 x 100%
2125,1 mg
= 17,17%
Fat
Sample
Weight
(g)
Flask (g) Boiling Chip (g)
Final
Weight
(g)
% Fat Average
1 5,0051 105,9132 3,1861 1,5591 31,15
30,97
2 5,0177 106,6245 1,5151 1,5449 30,79
Calculation:
% Fat =
weight of fat extracted (g)
weight of sample (g)
× 100%
= 1,5591 x 100%
5,0051
= 31,15%
Ash
Sample
Weight
(g) Crucible (g) Final Weight (g) % Ash Average
1 5,0345 20,9339 0,0790 1,57
1,66
2 5,0122 29,2951 0,0877 1,75
Calculation:
% Ash content =
x−y
z
× 100%
= (21,0129 - 20,9339) x 100%
5,0345
= 1,57%
Moisture
Sample Weight Crucible (g) Final Weight(g) % Moisture Content Average
1 5,1224 39,0469 4,7136 7,98
8,25
2 5,1883 41,1139 4,7463 8,52
Calculation:
% Moisture content =
W1−W2
W1
X 100%
= 5,1224 - 4,7136 x 100%
5,1224
= 7,98 %
Carbohydrate
No
Carbohydrate Content
(%)
Average (%)
1 42,13
41,88
2 41,63

A-2
Calculation:
% Carbohydrate = 100% - %Moisture - %Ash - %Fat - %Protein
= 100% - 8,25% - 1,66% - 30,97% - 17,24%
= 41,88%

A-3
Fiber and Omega-3 Fatty Acid Content of Chia Seed

B-1
Appendix B. Questionnaire of Comparison Test
SIMPLE COMPARISON TEST
Product : Cookies
Name : Date :
Cicipi sampel dari kiri ke kanan. Jangan mencicipi ulang sampel. Tulis kode dari sampel yang
lebih keras.Bilas mulut dengan air sebelum mencicipi sampel berikutnya.
Code
SIMPLE PAIRED PREFERENCE TEST
Product : Cookies
Name : Date:
Cicipi sampel dari kiri ke kanan. Jangan mencicipi ulang sampel. Tulis kode dari sampel yang
lebih disukai. Bilas mulut dengan air sebelum mencicipi sampel berikutnya.
Code

C-1
Appendix C. Result of Comparison Test
Simple Comparison Test
No Answer
40 One-stage
41 One-stage
42 One-stage
43 One-stage
44 One-stage
45 One-stage
46 One-stage
47 One-stage
48 One-stage
49 One-stage
50 One-stage
51 Two-stage
52 One-stage
53 One-stage
54 One-stage
55 One-stage
56 One-stage
57 One-stage
58 One-stage
59 One-stage
60 One-stage
61 One-stage
62 One-stage
63 Two-stage
64 Two-stage
65 One-stage
66 One-stage
67 One-stage
68 One-stage
69 One-stage
70 One-stage
71 One-stage
72 One-stage
73 One-stage
74 One-stage
75 One-stage
No Answer
1 One-stage
2 One-stage
3 One-stage
4 One-stage
5 One-stage
6 One-stage
7 One-stage
8 One-stage
9 One-stage
10 One-stage
11 One-stage
12 One-stage
13 One-stage
14 One-stage
15 One-stage
16 One-stage
17 One-stage
18 One-stage
19 One-stage
20 One-stage
21 One-stage
22 One-stage
23 One-stage
24 One-stage
25 One-stage
26 One-stage
27 One-stage
28 One-stage
29 One-stage
30 One-stage
31 One-stage
32 One-stage
33 One-stage
34 One-stage
35 One-stage
36 One-stage
37 One-stage
38 One-stage
39 One-stage

C-2
Simple Paired Preference Test
No Answer
40 Two-stage
41 One-stage
42 Two-stage
43 Two-stage
44 Two-stage
45 Two-stage
46 Two-stage
47 Two-stage
48 Two-stage
49 Two-stage
50 Two-stage
51 Two-stage
52 Two-stage
53 Two-stage
54 Two-stage
55 Two-stage
56 Two-stage
57 Two-stage
58 Two-stage
59 Two-stage
60 Two-stage
61 Two-stage
62 Two-stage
63 Two-stage
64 One-stage
65 One-stage
66 One-stage
67 Two-stage
68 Two-stage
69 Two-stage
70 Two-stage
71 Two-stage
72 Two-stage
73 Two-stage
74 Two-stage
75 Two-stage
No Answer
1 Two-stage
2 One-stage
3 Two-stage
4 Two-stage
5 Two-stage
6 One-stage
7 Two-stage
8 Two-stage
9 Two-stage
10 Two-stage
11 One-stage
12 Two-stage
13 Two-stage
14 Two-stage
15 One-stage
16 Two-stage
17 Two-stage
18 One-stage
19 Two-stage
20 One-stage
21 Two-stage
22 Two-stage
23 One-stage
24 Two-stage
25 Two-stage
26 Two-stage
27 Two-stage
28 Two-stage
29 Two-stage
30 Two-stage
31 Two-stage
32 One-stage
33 Two-stage
34 Two-stage
35 Two-stage
36 Two-stage
37 Two-stage
38 Two-stage
39 Two-stage

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