EFFECT OF FORMULATION, ENDPOINT TEMPERATURE,

AND HYDROCOLLOIDS ON QUALITY CHARACTERISTICS

OF CHICKEN LIVER SAUSAGES

Undergraduate Thesis

DENNIS GUIDO
203135030033497

FOOD TECHNOLOGY STUDY PROGRAM

FACULTY OF LIFE SCIENCES
SURYA UNIVERSITY
TANGERANG
2017
STATEMENT OF THESIS AUTHENTICITY

I, hereby undersigned,
Name: Dennis Guido
Student ID Number: 203135030033497
Major Program: Food Technology
Thesis Title: Effect of Formulation, Endpoint Temperature, and Hydrocolloids on
Quality Characteristics of Chicken Liver Sausages
affirm that my thesis is an original piece of work, written, and completed on my
own, and haven’t been published. The thesis was based on data observation,
reference books, journals, and other sources as listed on the reference section,
following the guideline from Surya University.
In accordance to Permendiknas Republik Indonesia Nomor 17 Tahun 2010
tentang Pencegahan dan Penanggulangan Plagiat di Perguruan Tinggi, if my
statement above is proven untrue, I am ready to accept the sanctions and
consequences to my bachelor degree revocation that is decided by Surya
University.
Tangerang, 13 June 2017
Materai 6000

Dennis Guido

i
 ABSTRACT

DENNIS GUIDO. Effect of Formulation, Endpoint Temperature, and

Hydrocolloids on Quality Characteristics of Chicken Liver Sausages. Supervised
by Astya R. Nilamsari and Saowakon Wattanachant.

Sausage is an ideal product to be added by liver among other processed meat

products, because it could be made by many formulations and in many forms. This
study was aimed to develop good textural properties of chicken liver sausages
(CLS). Variances of chicken liver, meat, and fat ratio and different contents of
hydrocolloids were formulated by Design Expert 7.0 software using Mixture
Design to produce CLS. Different endpoint temperatures (65oC, 75 oC, 85 oC) on
quality of CLS were also examined to determine the optimal condition for cooking
products. Formula optimization resulted that sausage with meat content of 33%,
liver content of 12%, and fat content of 15% had the highest desirability value
(0.809). Desirability value approaching 1 indicated a better result. Cooking CLS to
85oC endpoint temperature had produced sausage with higher value of hardness,
cohesiveness, and chewiness (p<0.05). Different ratios of hydrocolloids (Iota-
carrageenan, sodium alginate, and tapioca starch) had a significant impact on
texture profile, color values (a* and b*), cooking loss, and expressible moisture of
the product. Sample with carrageenan level of 4.5%, alginate level of 0%, and
starch level of 0% had the highest sensory score in texture, appearance, and overall
preference (p<0.05). CLS with optimized formulation obtained the highest
desirability value (0.846).

Keywords: Chicken liver, Design Expert 7.0, Liver sausages, Hydrocolloids

ABSTRAK

Sosis merupakan produk ideal yang dapat ditambahkan produk sampingan seperti
hati ayam dibandingkan produk olahan daging lainnya. Penelitian ini bertujuan
untuk mengembangkan karateristik tekstur yang baik pada sosis hati ayam (SH).
Variasi rasio dari daging ayam, hati, dan lemak, juga kadar hidrokoloid yang
berbeda diformulasikan menggunakan peranti lunak Design Expert 7.0
menggunakan fitur Mixture Design untuk memproduksi SH. Suhu titik akhir yang
berbeda (65oC, 75 oC, 85 oC) pada kualitas SH juga dipelajari untuk menentukan
kondisi optimal ketika memasak produk. Hasil dari optimasi formula
memperlihatkan bahwa sosis dengan kadar daging 33%, hati 12%, dan lemak 15%
mendapatkan nilai desirability tertinggi (0.809). Nilai desirability mendekati 1
menunjukkan hasil yang lebih baik. Sosis yang dimasak hingga suhu titik akhir
85oC akan menghasilkan produk yang memiliki hardness, cohesiveness, chewiness,
dan lightness lebih tinggi (p<0.05). Formulasi hidrokoloid (Iota-karagenan,
sodium alginat, dan pati tapioka) memiliki pengaruh signifikan terhadap profil
tekstur, warna (a* dan b*), susut masak dan expressible moisture. Sampel dengan
kadar karagenan 4.5 %, alginat 0% dan pati 0% memiliki skor hedonik tertinggi
pada parameter tekstur, penampakan, dan keseluruhan (p<0.05). SH dengan
formula optimasi mendapatkan nilai desirability tertinggi (0.846).

Kata kunci: Design Expert 7.0, Hati ayam, Hidrokoloid, Sosis hati

ii
 EFFECT OF FORMULATIONS, ENDPOINT
TEMPERATURES, AND HYDROCOLLOIDS ON QUALITY
CHARACTERISTICS OF CHICKEN LIVER SAUSAGES

Submitted to fulfill one of the requirements

to acquire Sarjana Teknologi Pertanian degree
on Food Technology Study Program

DENNIS GUIDO
203135030033497

FOOD TECHNOLOGY STUDY PROGRAM

FACULTY OF LIFE SCIENCES
SURYA UNIVERSITY
TANGERANG
2017

iii
 APPROVAL BY THESIS SUPERVISOR

Thesis Title : Effect of Formulation, Endpoint Temperature, and Hydrocolloids

on Quality Characteristics of Chicken Liver Sausages
Name : Dennis Guido
Student ID : 203135030033497

has submitted the thesis as part of requirements to acquire a Sarjana Teknologi

Pertanian degree in the Nutrition and Food Technology Program, Faculty of Life
Sciences, Surya University.

Approved by:

Supervisor 1: Astya Rizki Nilamsari, M.Si ( )

Supervisor 2: Assist Prof. Dr. Saowakon Wattanachant ( )

Acknowledged by:
Acting Head of Food Technology Study Program

Dr. Eng. Niki Prastomo

iv
 APPROVAL BY THESIS EXAMINING COMMITTEE

Thesis Title : Effect of Formulation, Endpoint Temperature, and Hydrocolloids on

Quality Characteristics of Chicken Liver Sausages’
Name : Dennis Guido
Student ID : 203135030033497

has submitted the thesis report as part of requirements to acquire a Sarjana Teknologi
Pertanian degree in the Nutrition and Food Technology Program, Faculty of Life
Sciences, Surya University.

Approved by:

Examining Committee 1: Tina Nurkhoeriyati S.TP, M.Sc ( )

Examining Committee 2: Dr. Ing. Dase Hunaefi S.TP, M.Sc ( )

Acknowledged by:
Acting Head of Food Technology Study Program

Dr. Eng. Niki Prastomo

v
 ACKNOWLEDGEMENT

Praise to the Lord so that author could successfully finished this thesis report.
Writer had chosen topic entitled ‘Effect of Formulation, Endpoint Temperature, and
Hydrocolloids on Quality Characteristics of Chicken Liver Sausages’ which research
was conducted since January 2017 until May 2017 in Prince of Songkla University,
Hatyai, Thailand.
Author would like to express his gratitude for :
1. Prince of Songkla University (PSU), Hatyai, for funding and giving chance to
Indonesia students, thus we could have conducted our research internship.
2. Surya University, for letting students to had research internship abroad in
Thailand.
3. Asst. Prof. Dr. Saowakon Wattanachant as supervisor in Thailand for kindly
guiding and expertly giving advice for this project.
4. Astya Rizki Nilamsari, M.Si as supervisor in Indonesia for her advice on this
project writing and seminar.
5. Pensiri Koi for guiding author in doing lab work and giving advice for the
project.
6. Sirinthip Saelin and Piyamon Duangchuay for assisting author on purchasing
raw ingredients and lab work.
7. Lab 2219 members for their friendly support in doing project.
8. All friends from PSU who contributed as panelists of sensory evaluation and
also made cheerful mood for author on working this project.
Hopefully this project would be beneficial for reader.

Hat Yai, May 2017

Dennis Guido
203135030033497

vi
 TABLE OF CONTENTS

STATEMENT OF THESIS AUTHENTICITY i

ABSTRACT ii
APPROVAL BY THESIS SUPERVISOR iv
APPROVAL BY THESIS EXAMINING COMMITTEE v
ACKNOWLEDGEMENT vi
TABLE OF CONTENTS vi
LIST OF FIGURES ix
LIST OF TABLES x
LIST OF APPENDIXES xi

CHAPTER I INTRODUCTION 1
1.1. Background of the study 1
1.2. Statements of the Problem 3
1.3. Objectives of the Study 3
1.4. Significance of the Study 3
1.5. Scope of Study 3

CHAPTER II LITERATURE REVIEW 5

2.1. Chicken Liver 5
2.2. Liver Sausage 6
2.3. Additives on Liver Sausages 7
2.4. Sausage Casing 8
2.5. Carrageenan 9
2.6. Alginates 9
2.7. Tapioca Starch 10
2.8. Texture Profile Analysis (TPA) 11
2.9 Design Expert Software 12

CHAPTER III METHODOLOGY 14

3.1. Time and Place of Study 14
3.2. Materials & Instruments 14
3.3. Research Design 15
3.3.1. Part 1: Determining the Optimal Ratio of Chicken Liver, Meat, and Fat to
Produce Chicken Liver Sausage 15
3.3.2. Part 2: Effect of Different Endpoint Temperature on Chicken Liver
Sausage 17

viii
 3.3.3. Part 3: Determining the Optimal Ratio of Different Hydrocolloids Added to
Chicken Liver Sausage 18

CHAPTER IV RESULT & DISCUSSION 21

4.1. Part 1: Determining the Optimal Ratio of Chicken Liver, Meat, and Fat to
Produce Chicken Liver Sausage 21
4.1.1. Proximate Analysis 21
4.1.2. Expressible Moisture 21
4.1.3. TPA 22
4.1.4. pH 23
4.1.5. Formula optimization 24
4.2. Part 2: Effect of Different Endpoint Temperature on Chicken Liver Sausage 25
4.3. Part 3: Determining the Optimal Ratio of Different Hydrocolloids Added to
Chicken Liver Sausage 27
4.3.1. Texture Profile Analysis (TPA) 27
4.3.2. Color Analysis (Hunter LAB) 29
4.3.3. Expressible Moisture and Cooking Loss 31
4.3.4. Appearance of Products 33
4.3.5. Sensory Evaluation 33
4.3.6. Formula optimization 35

CHAPTER V CONCLUSION 36
5.1 Conclusion 36
5.2 Suggestion 37

REFERENCES 38
APPENDIXES 42

viii
 List of Figures
Figure 1. Texturometer curve 12
Figure 2 Simplex plot of part 1 formulation by DX 7.0 16
Figure 3 Simplex plot of part 3 formulation by DX 7.0 18
Figure 4. Expressible moisture value on chicken liver sausages 22
Figure 5. Hardness value on chicken liver sausages 23
Figure 6. Cohesiveness value on chicken liver sausages 23
Figure 7. Chewiness value on chicken liver sausages (kg) 23
Figure 8. pH value of chicken-liver sausages 24
Figure 9. Texture profile analysis of chicken-liver sausage 28
Figure 10 Contour graphs of TPA results 29
Figure 11. Model graph of color analysis (L*, a*, b*) results 30
Figure 12. Contour graphs of expressible moisture and cooking loss 32
Figure 13. Appearance of Chicken-liver sausage samples 33
Figure 14. Sensory scores of selected samples 34

x
 List of Tables
Table 1. Nutritional Value of Chicken, Beef and Pork Liver 5
Table 2. Amylose–Amylopectin ratios for some starches 11
Table 3. Ratio of chicken liver, meat, and fat to produce chicken liver sausage (%) 16
Table 4. Ratio of Hydrocolloids Added to Chicken Liver Sausage (%) 19
Table 5. Proximate results of chicken liver and breast meat 21
Table 6. Criteria of optimization on ratio of meat, liver, and fat 24
Table 7. Solutions with desirability value from DX software 25
Table 8. Texture profile analysis of chicken liver sausage on different endpoint temperature
compared to the commercial 26
Table 9. Color values of chicken liver sausage on different endpoint temperature 26
Table 10. Expressible moisture of chicken liver sausage on different endpoint temperature
compared to the commercial 27
Table 11. Color value from chicken-liver sausage 30
Table 12. Cooking loss and expressible moisture of chicken-liver sausage 31
Table 13. Criteria of optimization on ratio of hydrocolloids 35
Table 14. Solutions with desirability value from DX software (%) 35

x
 List of Appendixes
Appendix 1. Springiness value graph on chicken liver sausages with different ratio of
liver,meat and fat 42
Appendix 2. ANOVA statistical result of expressible moisture percentage on part 1 42
Appendix 3. ANOVA statistical result of hardness value on part 1 43
Appendix 4. ANOVA statistical result of springiness value on part 1 43
Appendix 5. ANOVA statistical result of cohesiveness value on part 1 44
Appendix 6. ANOVA statistical result of chewiness value on part 1 44
Appendix 7. ANOVA statistical result of hardness value on part 3 45
Appendix 8. ANOVA statistical result of springiness value on part 3 45
Appendix 9. ANOVA statistical result of cohesiveness value on part 3 45
Appendix 10. ANOVA statistical result of chewiness value on part 3 46
Appendix 11. ANOVA statistical result of lightness value on part 3 46
Appendix 12. ANOVA statistical result of redness value on part 3 46
Appendix 13. ANOVA statistical result of yellowness value on part 3 46
Appendix 14. ANOVA statistical result of cooking loss percentage on part 3 47
Appendix 15. ANOVA statistical result of expressible moisture percentage on part 3 47

x
 CHAPTER I
INTRODUCTION

1.1. Background of the study

In 2013, according to ASEAN Statistical Yearbook (2015), total production

of poultry meat in South East Asia was 7,786.4 thousand metric tons. Compared to
the previous years, production of poultry meat has increased each year from 6,759.8
thousand metric tons in 2010. Beside production of its meats, livers could also be
taken as by-product to be sold, eaten, and processed to some products. Production
of broilers in Indonesia has increased each year by 17.8% from 2010 (Indonesian
Minister of Agriculture, 2015). By the total production number, the manufactures
have a good potential to introduce chicken liver as processed products.
Consumption of liver in Indonesia per capita in 2014 was 0.104 kg. (Minister of
Agriculture, 2015). In order to increase selling values or shelf life of livers, it could
be processed into processed products including sausages, pâté, and others.
Independent survey by one of private company reported that consumption of
sausage by citizen in Indonesia had grown averagely 4.46% on each year (Herlina,
Darmawan, & Rusdianto, 2015).
There are limited numbers of research performed about chicken liver
processed products. In several countries, raw chicken livers are usually cooked and
eaten as side dish. In United States, it is regulated that variety meats (livers, tongue,
blood, etc) could only be used in cooked sausage, like hotdogs, as long as the name
of product is added by “with by-product” or “with variety meats” (Mohand, 2014).
Livers are added uncooked to the mixture of sausage to improving fat- and water-
binding ability in processed liver products (FAO, 2007). One of the liver processed
product is liver pâté, which made from a mixture of cooked ground meat and livers
added with fat, minced into a spreadable paste. Sausages and pâté is an emulsion,
which quality linked to a good mixing of good mixing of fat, water and soluble
protein. Hamzeh et al (2016), explained that chicken liver is the top of other liver
types (calf, sheep) in increasing pH value to liver pâté. The pH value is an essential
factor during the emulsification process, and effect on the functional and physic-
chemical properties of the emulsion (Feiner, 2006). They also advised to prepare

1
liver pâté with three liver types (calf, sheep, chicken) and 40/10 % fat/water, which
led to the higher stability of emulsion.
Sausage is an ideal meat product, it could be made by many formulations
and in many sizes (Puolanne, 2010). Production of liver sausages is a method that
maximizes the utilization of unpopular raw meats such as organ meats and
byproducts and converts these less expensive raw materials into highly nutritious
products (Chyr, 1978). Formulation of sausage-making requires an assemblage of
various ingredients in the right proportion to produce not just a desired quality and
safe product but also a cost-effective one (Essien, 2003). One of the key factors of
sausage in meeting customer’s requirement is texture of the product (Dincer &
Çakli, 2015). Textural properties in sausage depends on the structure of the proteins
gel matrix, the soluble matters and moisture which were entrapped in the gels
structure. To improve the texture, texturizing agents like hydrocolloids often used
as gel-forming in sausages (Nicomrat et al., 2015). Petrachi and Bianchi (2012)
stated that among the many hydrocolloids available in the market, carrageenan and
alginates are the most commonly used in poultry meat products formulation. Also,
Nicomrat et al (2015) showed that iota carrageenan and sodium alginate will be
suitable texturing agents for improving the texturing of sausage. Moreover, the
sausage by the addition of sodium alginate can reduce cooking loss and expressible
moisture
On this study, different ratio of chicken liver, chicken meat, and fat were
proposed to make chicken liver sausages assisted by Design Expert 7.0 software.
Design Expert is a statistical software package from Stat-Ease Inc, which is
specifically dedicated to performing design of experiments (DOE). The optimal
formulas will be used to study the effect of different endpoint temperatures (65oC,
75 oC, 85 oC) on cooking chicken-liver sausages. The research will proceed by
adding hydrocolloids such as iota-Carrageenan (Cr), Sodium Alginate (A), and
Tapioca Starch (S) to produce chicken liver sausages. The samples will be
examined with physical, physicochemical, color and sensory evaluation to
determine the optimal ratio to produce chicken liver sausage.

2
1.2. Statements of the Problem

1. Which is the optimal ratio of chicken liver, chicken meats, and fat to produce
chicken liver sausage with high textural properties?
2. What is the effect of different endpoint temperatures on cooking chicken-liver
sausage?
3. How are textural, physicochemical properties and color of chicken liver
sausages added with different hydrocolloids?
4. Which is the optimal ratio of iota-carrageenan, sodium alginate, and tapioca
starch to produce chicken liver sausage?

1.3. Objectives of the Study

1. To determine the optimal ratio of chicken liver, chicken meats, and fat to
produce chicken liver sausage with high textural properties
2. To examine effect of different endpoint temperatures on cooking chicken-liver
sausage.
3. To examine textural properties and color of chicken liver sausages added with
different hydrocolloids
4. To determine the optimal ratio of iota-carrageenan, sodium alginate, and tapioca
starch to produce chicken liver sausage.

1.4. Significance of the Study

1. The result of the study could suggest the optimal ratio of chicken liver, chicken
meats, and fat to produce chicken liver sausage with high textural properties.
2. The study was expected to report effect of different endpoint temperatures on
cooking chicken-liver sausage
3. The result of the study could suggest the best ratio of iota-carrageenan, sodium
alginate, and tapioca starch for making chicken liver sausage.

1.5. Scope of Study

This study was focused on producing the higher textural properties of

chicken-liver sausages. Design of formulas were proposed by Design Expert

3
software to assist author in determining the optimal formula. Variances of chicken
liver, meat, and fat ratio and different hydrocolloids ratio were used to produce
chicken liver sausage. Different endpoint temperatures on chicken liver sausages
also examined to determine the optimal endpoint temperature on cooking
products.

4
 CHAPTER II
LITERATURE REVIEW

2.1. Chicken Liver

Gallus domesticus or chicken is the species of domesticated bird. Chicken
liver is one of the items to be traded, else whole breast, thigh, and wings (Legarreta,
2010). Liver and other various organs, included tongue, heart, kidney, spleen,
brains, retina, intestines, tripe, etc. are called variety meats. Many of these variety
meats, such as liver, kidney or heart, are highly-valued foods because they contain

Table 1. Nutritional Value of Chicken, Beef and Pork Liver

liver/ 100g unit Chicken Beef Pork
Water g 76.46 70.81 71.06
Energy kcal 119 135 134
Proximates Protein g 16.92 20.36 21.39
Total Lipid g 4.83 3.63 3.65
Carbohydrate g 0.73 3.89 2.47
Calcium mg 8 5 9
Iron mg 8.99 4.9 23.3
Magnesium mg 19 18 18
Mineral Phosphorus mg 297 387 288
Potassium mg 230 313 273
Sodium mg 71 69 87
Zinc mg 2.67 4 5.76
Vitamin C mg 17.9 1.3 25.3
Thiamin mg 0.305 0.189 0.283
Riboflavin mg 1.778 2.755 3.005
Niacin mg 9.728 13.175 15.301
Vitamin B6 mg 0.853 1.083 0.609
Vitamins Folate µg 588 290 212
Vitamin B12 µg 16.58 59.3 26
Vitamin A, IU IU 11078 16898 21650
Vitamin E mg 0.7 0.38 -
Vitamin D IU - 49 -
Vitamin K µg - 3.1 -
Total Saturated g 1.563 1.233 1.17
Total Monounsaturated g 1.249 0.479 0.52
Lipids Total Polyunsaturated g 1.306 0.465 0.87
Total Trans g 0.065 0.17 -
Cholesterol mg 345 275 301
Source: US Agricultural Research Service (2016)

5
vitamins and trace elements as well as high quality protein (Grosch, Belitz, &
Schieberle, 2009). Liver function in animal body is to absorb most of the nutrient
from food and are either stored or processed for distribution to other organs
(McGee, 2004).
Because of its high blood content, liver must be boiled until it is at least over
half-cooked, using special equipment reserved for this purpose. It cannot be soaked
and salted. The liver is then rinsed, after which the liver can be used in any way that
the user wishes. Enough salt must be sprinkled on the liver to cover it (Legarreta,
2010). Without the hygienical handling, liver has been found as a cause of
Campylobacteriosis outbreaks (Hutchison, Harrison, Richardson, & Tchórzewska,
2015). It has been determined that more than 70 °C for at least two minutes at a
liver’s core must be attained to cook livers safely (Whyte, Hudson, & Graham,
2006).
If liver were cooked for too long, they become gray and unappetizing with
changes in texture. Hutchison et al (2015) confirmed that cooking to 63 oC is a
critical control point to destroy Campylobacter with bain-marie, a container holding
hot water which a pan is placed for slow cooking.

2.2.Liver Sausage
Liver sausage or called Leberwurst in Germany, is a typical sausage served
in Germany, Hungary, the Netherlands, Finland, and Sweden. Most liver sausage
varieties are spreadable. It is usually made with pork. Liver added to the sausage
product is only about 10-20%, which is enough to give it a typical livery taste. Other
ingredients are meat, fat, and spices, such as ground black pepper, marjoram,
allspice, thyme, ground mustard, or nutmeg (Puolanne, 2010). Liver sausage
categorized as emulsion sausage by Essien (2003), other examples are bologna,
kochwurst, and frankfurters. The varieties of liver sausage are endless because all
possible spices and herbs as well as other materials such as wine and brandy can be
used as flavoring materials. Commonly, raw meat and fat used in manufacture of
liver sausage and raw liver is used as emulsifier. Liver sausage generally is cured
and nitrite is added. Liver is added to give impact on taste, the emulsification of fat
and water, and to stabilize the sausage. Pork is generally used in liver sausage but
chicken or beef can also be used without any problems as well (Feiner, 2006).

6
 Raw materials of liver sausage should get through selection and preparation.
A low bacteria count on meats used in liver sausage is desirable and 102 and 104 cfu
per gram of product is the optimum. A large number of bacteria would give negative
impact to the finished product on parameters such as color development, flavor, and
color stability. Liver sausages commonly contains a fat content between 25% and
40 %. The texture of a liver sausage is mostly affected by solidified fat, which is
covered in a protein matrix made from solubilized liver protein as well as gelatin.
A level of fat between 20% and 40% can be present within liver sausage without
the risk of fat separation if the amount of liver within the product is between 25% -
30% as well. Liver sausages can be produced with oil instead of using fat. Oil is
used in products such as beef or chicken liver sausage (Feiner, 2006).

2.3. Additives on Liver Sausages

The variety of additives added to the product vary significantly. Liver
sausages is mildly salted and the amount of salt is commonly between 12 g to 18 g
per kilogram of product. Salt is added for its contribution to taste and flavor. The
presence of salt reduces aw value of the product but not significance to parameter
such as shelf life as liver sausages has a high water activity by nature (Feiner, 2006).
It is essential to consider that when formulating, the level of sodium in the final
product will be influenced by salt added directly at the mixing point and that from
other ingredients such as rusk, gel bind, cure compound and seasoning mix (Essien,
2003). Salt should be pure and finely granulated so it can dissolve easily in the meat.
Spices added to sausage are found in many forms: fresh, dried, whole,
crushed, pureed, as paste and as an extract. Most consumers prefer the taste of
freshly made food that often comes from spices (Mohand, 2014). Sage, parsley,
coriander, paprika, rosemary, nutmeg, leek, apricot, cranberry, tomato, stilton,
onions and mint are ingredients commonly used as flavorings, spices and herbs in
sausages (Essien, 2003). Spices and herbs added according to taste. In liver
sausages, fresh onions are not used owing to their high glycogen content and
occasionally high bacteria count. The glycogen content in onions combined with
the glycogen present in liver could make a sour taste in the finished product. Onion
powder is microbiologically much safer (Feiner, 2006).

7
 Emulsifier such as monoglyceride and diglyceride of fatty acids and other
are frequently used in liver sausage to reduce the risk of fat separation during
thermal processing. Monoglycerides have stronger hydrophilic tendencies than
diglycerides which are more lipophilic. The quantity of emulsifier used in general
is 3-5 g per kilogram of sausage mass. Other ingredients, such as sugars, are
commonly mixed with the emulsifying agent. Monoglyceride containing around
25% citric acid seem to reduce the risk of fat separation in pasteurized products.
Natural emulsifiers such as caseinate, egg protein and blood plasma also stabilize
an emulsion in the products. Only egg protein forms a matrix as liver protein does
(Feiner, 2006).
2.4. Sausage Casing
Casing, or called skins are divided into two types, natural and artificial. It is
used in sausage manufacture to achieve primary significance in portioning (Essien,
2003). Natural casings are derived from the digestive tract of animals. These
casings are made from sheep (3/4 inches), hog (1-3/8 inches) and cattle (1-3/8
inches) intestines. Natural casings always have natural curve on them and desirable
‘snap’ when it is bitten (Mohand, 2014). To avoid microbial contamination, natural
casings should be stored at temperature below 4.5 oC. The use of natural casing is
not popular because of handing problems, splitting and difficulty in standardizing
the weight and dimensions of sausage.
Artificial casings are made with collagen, cellulose, fibrous and plastic
materials to suit a wide range of applications. Collagen is extracted from connective
tissue of animals and used as sausage casing (Essien, 2003). Cellulose casing is
made from inedible plant and tree fibers. Casings from cellulose allow smoke to
penetrate and moisture to escape during cooking. It has the advantage of uniform
in size (diameter) and free of defects (Mohand, 2014). Fibrous casings are
manufactured by extruding regenerated cellulose onto a paper base and forming
into tubes. This produces a strong, inedible container for large diameter (2-6 inches)
products. This product is especially useful for water-cooked items such as bologna,
liver sausage, and poultry rolls. These casings are typically removed prior to slicing,
portioning, or packaging (Keeton, 2001).

8
 Beside from providing shape to sausage, casings also have an impact on
increasing product shelf life by providing high moisture and oxygen resistance
properties with seal strength and density. Therefore, also contribute in minimizing
weight loss of product during cooking. When choosing types of casing, a balance
between high resistance to splitting coupled with easy dissolution in the mouth
while eating should be the prime considerations (Essien, 2003).

2.5. Carrageenan
Carrageenan is sulfated galactopyronase polysaccharide from Rhodophyta
family which contains methoxy and pyruvate group. There are three types of
carrageenan depends on number and position of sulphate groups in 3,6-anhidro
bridge structure, such as iota, kappa and lambda carrageenan (Feiner, 2006).
Experiment by Nicomrat et al (2015) shown that iota carrageenan decreased more
cooking loss and expressible moisture content compared to kappa carrageenan.
Candogan & Kolsarici in Nicomrat et al (2006) explained that with increasing
carrageenan concentration, WHC increased and penetrometer value decreased in
low-fat frankfurters. Moreover, Ayadi et al (2008) reported that addition of
carrageenan at 0.2 % and 0.5 % increases gel elasticity. Higher carrageenan
concentration reduces sausages elasticity. By sensorial analysis, it shows that
carrageenan presence has no significant effect on sausages taste. However, it
improves sausage appearance and texture.

2.6. Alginates
Alginate was first described by the British chemist Stanford in 1881.
Alginate is the salt of alginic acid that is a gum-like derivate from brown seaweed
Macrocystis pyrifera. (Feiner, 2006). It is located in the intercellular matrix as a gel
containing sodium, calcium, magnesium, strontium, and barium ions. Its main
function is to be skeletal, which giving both strength and flexibility to the algal
tissue. Alginate is widely used industrially because of its ability to retain water and
its gelling, viscosifying and stabilizing properties.
The first step of its production is the insoluble alginate with a counterion
composition determined by the ion exchange equilibrium with seawater is ion-
exchanged with protons by extracting the milled algal tissue with 0.1 – 0.2 M

9
mineral acid. The second step is brought the alginic acid into solution by
neutralization with alkali such as sodium carbonate or sodium hydroxide to form
water-soluble sodium alginate. After removal of algal particles by separation
procedures like sifting, flotation, centrifugation, and filtration, the product is
precipitated by addition of alcohol, calcium chloride, or mineral acid, reconverted
to the sodium form and finally dried and milled (Draget, Moe, Braek, & Smidsrod,
2006).
Alginates can be used as meat replacers whereby a little amount of meat is
combined with a high amount of water plus flavors and colorants to yield a gelled
alginate matrix that can be subsequently used as a raw material for a variety of low-
value meat products. Applications of alginate preparations allow to produce stable
cold fat emulsions to be used for including a stable form of fat in product
formulations such as chicken nuggets or sausages (Petracci & Bianchi, 2012).
Addition of alginate to beef-sausage effectively improved water holding capacity
(WHC), cooking yield, and textural properties (El-Baki et al, 1981 in Santana et al,
2013). Experiment also done by Juemanee et al. (2009) which proved that addition
of alginate to patty had an impact to moisture content and juiciness scores.

2.7.Tapioca Starch
Tapioca starch is derived from the cassava plant. Names for the cassava
plant vary depending on the region: yucca (Central America), mandioca or manioc
(Brazil), tapioca (India and Malaysia) and cassada or cassava (Africa and Southeast
Asia). The plant belongs to the Euphoriaceae family. Previously, cassava was
described as two edible species of the genus Manihot, Manihot ultissima Phol and
Manihot palmata, based on the presence of high and low cyanide contents in roots.
Recently, both bitter and sweet cassava classes were classified being the same
species of Manihot esculenta. For starch, the bitter cassava type mostly used in the
production industry, because the sweet type is used for direct consumption
(Breuninger, Piyachomkwan, & Sriroth, 2009).

In meat products, starches are mainly used as gelling agents in minced meat
systems such as chicken nuggets by bind the water during cooking and subsequent
improvement of shelf-life, as well as deliver the desired texture. Starches are

10
polysaccharides which are formed by different amylose and amylopectin. Amylose
is mostly responsible for the firmness or gel strength of a starch gel. Amylopectin
is responsible for the elasticity and viscosity of a starch gel. Starches high in
amylopectin are easier to cook and generally gelatinize at lower temperatures than
starches high in amylose (Petracci & Bianchi, 2012). According to Table 2, ratio of
amylose and amylopectin of tapioca starch is about 1:5. It is on the same level of
ratio compared with rice starch, and cassava.

Table 2. Amylose–Amylopectin ratios for some starches

Starch Source Amylose (%) Amylopectin (%)
High Amylose Corn 50-85 15-50
Corn 26 74
Waxy corn 1 99
Wheat 25 75
Rice 17 83
Cassava 17 83
Tapioca 17 83
Potato 21 79
Source: Zobel & Stephen (2006)

2.8.Texture Profile Analysis (TPA)

Texture profile analysis is an empirical technique using a two-cycle
compression test that resembles the action of a jaw. This analysis was pioneered by
a group of food scientists from the General Foods Corporation. Data collected as
force during compression and time. Parameters such as hardness, brittleness,
cohesiveness and springiness can be analyzed from this test (Daubert & Foegeding,
2010). The principle of the TPA test is a sample of food placed on the baseplate and
compressed two times by a plate. To resemble the jaw action, there should be a high
compression on the sample. TPA is an important analysis on textural properties of
sausage, it also could quantify multiple textural parameters in just one experiment.

As illustrated above in a book by Bourne (2002), the height of the force peak
on the first compression cycle (first bite) was defined as hardness (A1). Brittleness
or fracturability was defined as the force of significant break in the curve on first
compression. The positive area of the curve (under A1 & A2) is called cohesiveness,
and the negative area is called adhesiveness. The distance between the time elapsed

11
 Figure 1. Texturometer curve

on the end of the first bite to the start of the second bite (BC) was defined as
springiness or called elasticity. Then gumminess was defined by calculation:
hardness x cohesiveness; chewiness was defined by calculation: gumminess x
springiness (Bourne, 2002).

2.9 Design Expert Software

Design Expert (DX) is a software that runs in a computer which is used to
optimize process or formulation of products. DX has 4 types on generating
experimental design such as Factorial Design, Combined Design, Mixture Design,
and Response Surface Methods (RSM) Design. Each of them has different function
and used as needed by the user. In the process of formulation, kind of options that
is used is Mixture Design, which determines optimization of formula from a series
components and materials added to the product (Hendy, 2007).
Generally, the steps in using DX are: (1) Design the composition of the
formula and determine the response to be tested; (2) Generating formula and
measuring responses on each formula; (3) Input the whole response data which is
analyzed on DX worksheet; (4) Significance analysis (ANOVA) and mathematic
model applied on each response and also determine the optimal formula to achieve
the goals of study.
Output of the experimental design by DX is series of formulas that has been
made and has each response analyzed. Determination of the optimal formula on
analyses step depending on the result of responses which are chosen by maximum,
minimum, in range, or any target options. The final result of the analysis is a new

12
formula which determined by the target established. The program will specify
several solutions with vary desirability value. The higher of the desirability value
(approaching 1) means the formula is getting more optimal (Hendy, 2007).

13
 CHAPTER III
METHODOLOGY

3.1. Time and Place of Study

The study was conducted at Lab of Meat and Poultry Sciences, Cooking
Lab, Food Analysis Lab, and Food Quality Lab, Faculty of Agro-Industry on
Prince of Songkla University (PSU), Hat Yai, Thailand from 4th January until
24th May 2017.

3.2. Materials & Instruments

3.2.1 Materials
Main materials used to produce sausage were chicken liver and chicken
breast meat acquired from Charoen Pokphand PCL, Thailand, also palm oil as
fat source from Morakot Industries Co., Ltd. Additives used on the product were
salt (Thai Refined Salt Co. Ltd, Thailand), Sodium Nitrite (Vidhyasom Co. Ltd,
Thailand) and Sodium Tripolyphosphate (Vidhyasom Co. Ltd, Thailand).
Sausage casing used was collagen casing acquired from Fujilin Thailand Co.,
Ltd , Thailand. Hydrocolloids used as texturizer agents were Iota-Carrageenan
(Modernist Pantry, New York), Sodium Alginate (Thai Food & Chemical Co.
Ltd, Thailand), and Tapioca Starch (Thai Food & Chemical Co. Ltd, Thailand).

3.2.2 Instruments
Instruments used on analyses of product were texture analyzer (Stable
Micro Systems TA.XT plus, England) to analyze TPA, centrifuge (Hitachi
CRI22G, Japan) to analyze expressible moisture, pH meter (Metler Toledo
8603, Switzerland) to analyze pH, colorimeter (Hunter LAB Miniscan EZ,
USA) to determine color values, water bath (Memmert GNB 7, Germany) to
determine cooking loss, and scale (Sartonius TE3102S & TE214S, Germany).

14
3.3. Research Design
3.3.1. Part 1: Determining the Optimal Ratio of Chicken Liver, Meat,
and Fat to Produce Chicken Liver Sausage
3.3.1.1. Proximate Analysis
Proximate Analysis (AOAC, 1999) of raw chicken liver and meat was
conducted to ensure that the contents of materials were consistent for each
formula.
3.3.1.1.1. Moisture Analysis
Test sample was weighed for 1-2 gram and dried in oven for 2-3 hours, then
cooled in desiccator for ca. 30 minutes. Moisture content was calculated based
on the difference in weight before and after drying.
3.3.1.1.2. Protein Analysis
Test sample was analyzed through 3 basic steps: 1) digestion of the sample
in sulfuric acid with a catalyst, which results in conversion of nitrogen to
ammonia; 2) distillation of the ammonia into a trapping solution; and 3)
quantification of the ammonia by titration with a standard solution.
3.3.1.1.3. Fat Analysis
Soluble material was extracted from dried test samples of meat and liver
with petroleum ether solution. Solvent is recovered by condensation, leaving
extracted soluble material. Fat content is determined by weight after drying
3.3.1.1.4. Ash Analysis
Meat and liver were weighed for 1-2 gram and then ignited at 600oC for 3
hours, cooled in desiccator for 30 minutes, and weighed soon after reaching
room temperature. Ash content was calculated based on the difference in weight
before and after igniting.
3.3.1.1.5. Carbohydrate Analysis
Total carbohydrate was calculated by difference as follows:
Carbohydrate (%) = 100% − (Moisture + Protein + Fat + Ash )% (3.1)
3.3.1.2. Formulation assisted by DX 7.0
Different ratio of chicken liver, chicken meat, and fat were proposed by DX
7.0 software and showed on Table 3. Constraints of each components were input
such as liver 12-24%, meat 18-33%, and fat 15-18%. Constraints were referred
to raw ingredients content range of liver sausage described by Feiner (2006).

15
Content of ingredients such as salt, ice and others were prepared according to
the method of Dianingtyas (2001) with modified from beef liver sausage to
chicken liver sausage. There were 10 formulas in result by Simplex Lattice
Design added with 3 formulas as replicates. Results from the analysis were
statistically processed with ANOVA then optimized by DX software.

Figure 2. Simplex plot of part 1 formulation by DX 7.0

Table 3. Ratio of chicken liver, meat, and fat to produce chicken liver sausage (%)
No Liver Fat Meat
1 24.0 17.0 19.0
2 17.2 16.4 26.5
3 17.9 18.0 24.1
4 24.0 17.0 19.0
5 22.1 15.0 22.9
6 17.2 16.4 26.5
7 17.2 16.4 26.5
8 12.0 15.0 33.0
9 14.3 18.0 27.7
10 19.8 15.3 24.9
11 12.0 17.6 30.4
12 15.0 15.0 30.0
13 20.1 18.0 21.9

3.3.1.3. Preparation of Chicken-Liver Sausage

Preparation began with cutting liver, and meats, into dice shape. Then liver
and meats were ground at the same time with grinder. Fat, salt (2.94 %), and
half of ice (16.25%) were added and ground for 1 minutes. Then carrageenan

16
and starch were added to batter with the remaining ice (16.25%), and ground
for 2 minutes. Batter was stuffed into collagen casing and cooked in water bath
at temperature 60oC for 30 minutes and then 80oC for 15 minutes.

3.3.1.4. Texture Profile Analysis

Texture Profile Analysis (TPA) of sample was determined according to the
procedure of Nicomrat et al (2015) by using Texture Analyzer (Stable Micro
Systems TA.XT plus, England) with 35 mm aluminum cylinder probe,
compressing for 50% original height. Sausage samples were left at room
temperature for 2 hours and then cut into 2 cm long cylinders.

3.3.1.5. pH analysis
Two grams of sausages were diluted in 10ml of water then analyzed by
digital pH meter.

3.3.1.6. Expressible Moisture

Expressible moisture of sample was determined according to the
procedure of Nicomrat et al (2015), 2 grams of sausages will be enveloped with
two pieces of Whatman no. 1 filter paper and centrifuged at 3,650 rpm for 15
min at 25o C. The expressible moisture content was calculated based on the
difference in weight before and after centrifuging.

3.3.2. Part 2: Effect of Different Endpoint Temperature on Chicken

Liver Sausage
3.3.2.1. Preparation of Chicken Liver Sausage
Chicken liver sausages were produced with determined formula from
previous part, then batter was separated into three portions. The first portion
was cooked to core temperature 65 oC, second portion was cooked to 75 oC,
and the third portion was cooked to 85 oC.. Core temperature was measured by
using piercing temperature probe.
3.3.2.2. Texture Profile Analysis
Methods of analyzing texture profile of sample as described on the
previous part (3.3.1.4)

17
3.3.2.3. Color Analysis
Color was analyzed as referred to the procedure of Jo et al (1999), using a
Hunter Colorimeter to measure A CIE Lab color [lightness (L*), redness (a*)
and yellowness (b*)] of sausages.

3.3.2.4. Expressible Moisture

Methods of analyzing expressible moisture of sample as described on the
previous part (3.3.1.8)

3.3.3. Part 3: Determining the Optimal Ratio of Different Hydrocolloids

Added to Chicken Liver Sausage
3.3.3.1 Formulation of Chicken Liver Sausage
On this research, variation of three different hydrocolloids (iota-
Carrageenan, Tapioca Starch, Sodium Alginate) ratio was designed by using
Simplex-Centroid Design and showed at Table 4. Simplex plot was plotted as
in Figure 3. Ratio of chicken liver, chicken meat, and fat to produce referred to
the optimal ratio from the previous part. Content of ingredients such as salt, ice
and others were referred to the method of Dianingtyas (2001) with modified
from beef liver sausage to chicken liver sausage. First ratio (1) was a control
variable which used in the previous study as no hydrocolloids added didn’t
resulted a sausage-like textural properties.

Figure 3. Simplex plot of part 3 formulation by DX 7.0

18
 Table 4. Ratio of Hydrocolloids Added to Chicken Liver Sausage (%)
No Carrageenan (Cr) Tapioca (T) Alginate (A)
1 2.5 2.0 0.0
2 0 0 4.5
3 1.5 1.5 1.5
4 0 2.25 2.25
5 2.25 2.25 0
6 4.5 0 0
7 0 4.5 0
8 2.25 0 2.25
9 0.75 3 0.75
10 3 0.75 0.75
11 0.75 0.75 3

3.3.3.2 Preparation of Chicken Liver Sausage

Preparation began with cutting liver (12%), and meats (33%), into dice
shape. Then liver and meat were ground at the same time with grinder. Fat
(15%), salt (2.94 %), and half of ice (16.25%) were added and ground for 1
minutes. Then hydrocolloids were added to batter with the remaining ice
(16.25%), and ground for 2 minutes. Batter was stuffed into collagen casing and
cooked in water bath at temperature 60oC for 30 minutes and then 80oC for 15
minutes.

3.3.3.3 Texture Profile Analysis

Methods of analyzing texture profile of sample as described on the previous
part (3.3.1.4)

3.3.3.4 Cooking Loss

Cooking loss value was determined according to the procedure of
Nicomrat et al (2015) by stuffing sausage batter to a 50 ml centrifuge tubes
about 40-45 grams and sealed with a plastic cap. The batter tubes were pre-
heated in water bath at 80oC for 30 min. Then removed from the hot water bath
and cooled in cold water at 5oC for 10 min. Cooking loss value was calculated
based on the difference in weight before and after heating.
3.3.3.5 Expressible Moisture
Methods of analyzing expressible moisture of sample as described on
the previous part (3.3.1.8)

19
3.3.3.6 Color Analysis
Methods of analyzing color values of sample as described on the
previous part (3.3.2.3)

3.3.3.7 Sensory Evaluation

Preference test by 30 panelists was conducted to 5 selected samples of
products to determine the most acceptable sample. Panelists invited were 12
males and 18 females of Bachelor, Master and Doctoral students from PSU at
age range of 21-30. Selection of samples were based on parameters such as
hardness, springiness, cohesiveness, chewiness, redness(a*), expressible
moisture, cooking loss, and appearance of samples (smoothness of the surface).
Sausage were cut in a half-round with 1 cm thickness and then score of color,
appearance, and texture were evaluated by panelists. To evaluate the texture of
products, panelists were instructed to dip the sample into tomato sauce and bite
slowly.

20
 CHAPTER IV
RESULTS & DISCUSSION

4.1. Part 1: Determining the Optimal Ratio of Chicken Liver, Meat, and
Fat to Produce Chicken Liver Sausage
4.1.1. Proximate Analysis
From Table 5, the results showed the proximate results of chicken liver
and breast meat acquired from Charoen Pokphand PCL, Thailand. Compared to
the results from Agricultural Research Service (2006) on Table 1, carbohydrates
level analyzed on liver were higher (3.79±0.88) than reference. Carbohydrates are
stored in the liver as glycogen (Legarreta, 2010). Protein in breast meat consisted
of connective tissue such as collagen and elastin, muscle protein such as actin and
myosin which act as gelling agent in sausage, also sarcoplasmic protein such as
albumin and globulin. While protein on liver mostly consisted of hemoglobin
(color of blood) which is one of the important types of globulin (Feiner, 2006).
Fat content of chicken liver is higher than other liver types (beef, pork) and mostly
contained unsaturated fat. On breast meat, fat mostly contained monounsaturated
fat (US Agricultural Research Service, 2006).

Table 5. Proximate results of chicken liver and breast meat

Content Liver Breast Meat
Moisture (%) 74.34±0.32 75.65±0.04
Protein (%) 16.90±0.56 18.54±0.99
Fat (%) 3.57±0.19 1.05±0.12
Ash (%) 1.41±0.09 1.08±0.06
Carbohydrate (%) 3.79±0.88 3.67±1.01

4.1.2. Expressible Moisture

As appeared in Figure 4, expressible moisture on samples were at range
34%-50%. Compared to the value reported by Nicomrat et al (2015), expressible
moisture on Moo yor sausage were at 15-20%. Moo yor is a popular Vietnamese
meat emulsion sausage. Expressible moisture was expected to be in a lower value
as it was contributed to desirable textural characteristics of sausage. After
statistically analyzed by DX, there were no model terms are significant on the

21
 variable, which indicated that expressible moisture value was not affected by the
formulation.
Meat was known as the major factor in decreasing expressible moisture, as
sausage with higher meat level would had higher salt soluble proteins, such as
actin and myosin. Actin and myosin as part of myofibrillar proteins act as gelling
proteins, playing a vital role in producing desirable textural characteristics in
processed muscle foods (Strasburg, Xiong, & Chiang, 2008).

60
a
Expressible Moisture (%)

ab abc
50
cd
cd d
bcd d d cd cd cd
40 d

30

20

10

0
1 2 3 4 5 6 7 8 9 10 11 12 13
Formula
Figure 4. Expressible moisture value on chicken liver sausages. Each value is presented
as mean ± standard deviation. Means above each bar with different letters differ
significantly (P<0.05)

4.1.3. TPA
On texture profile analysis (TPA), there were hardness, cohesiveness, and
chewiness that indicates model terms are significant. Variables which had
significant model terms could be used as criteria on optimization of formula. On
hardness showed in Figure 5, sample 8 with meat level 33%, liver 12%, and fat 15%
and sample 6 with meat level of 19%, liver level of 24%, and fat level of 17% had
the highest value amongst other samples. On cohesiveness showed in Figure 6,
sample 8 and sample 9 with meat level 27,7 %, liver 14,3%, and fat 18% had the
highest value amongst other samples. Lastly on chewiness showed at Figure 7,
sample 8 had the highest value. Chewiness value showed the higher value of
hardness, cohesiveness, and springiness respectively.
Youssef & Barbut in Nicomrat et al (2016) stated that texture parameters
could vary due to the formation of protein matrix and a rigid structure depending
on the amount of lean meat used in formulation. According to Cierach et al (2009),
the hardness in sausage is related to their fat content. These also could be related to

22
explanation by Feiner (2006) which said that the texture of liver sausage is largely
determined by solidified fat, covered in a protein matrix made from solubilized liver
protein as well as gelatin during thermal treatment. In low-fat liver sausages, water
tends to separate during thermal treatment, which liver level should be reduced and
replaced by meats.
4 a
ab bc bc bcd bcd
Hardness (kg)

3 bcd cde
de de de
ef
2 f
1

0
1 2 3 4 5 6 7 8 9 10 11 12 13
Formula
Figure 5. Hardness value on chicken liver sausages. Each value is presented as mean ±
standard deviation. Means above each bar with different letters differ significantly (P<0.05)

0.5
a
bc ab bc
Cohesiveness

0.4 bcd bcd

cd
0.3 e e de de
e e
0.2
0.1
0
1 2 3 4 5 6 7 8 9 10 11 12 13
Formula
Figure 6. Cohesiveness value on chicken liver sausages. Each value is presented as mean
± standard deviation. Means above each bar with different letters differ significantly
(P<0.05)
1.2
a
1 b
Chewiness (kg)

0.8 c
0.6 c cd
cd de cd
de de
0.4 de de
e
0.2
0
1 2 3 4 5 6 7 8 9 10 11 12 13
Formula
Figure 7. Chewiness value on chicken liver sausages (kg). Each value is presented as
mean ± standard deviation. Means above each bar with different letters differ
significantly (P<0.05)

4.1.4. pH
This analysis was conducted as pH could affect color, texture,
microbiological, and taste quality of sausage. Figure 8, it showed pH value of
chicken liver sausages at range 6.4 -7.0. pH value of sausage should not below 5.2,
as sol is transformed into a gel and could destroyed. If gel is destroyed, poor slice

23
coherency will be the result (Feiner, 2006). Model terms on this variable were not
significant which could be said that there is no effect by variances of ratio. This
could due to that addition or reduction of chicken liver, meat, and fat are not
affecting pH value of sausage product after cooking.

7.10
a
7.00 b bc bc c
6.90 d
6.80 d d
6.70 f e f
f
pH

6.60
g
6.50
6.40
6.30
6.20
6.10
1 2 3 4 5 6 7 8 9 10 11 12 13
Formula

Figure 8. pH value of chicken-liver sausages. Each value is presented as mean ± standard

deviation. Means above each bar with different letters differ significantly (P<0.05)

4.1.5. Formula optimization

As could be seen on Table 6, by aiming high textural properties of sausage,
significant variables such as hardness, chewiness and cohesiveness were used as
criteria of optimization process.

Table 6. Criteria of optimization on ratio of meat, liver, and fat

Name Goal Lower Limit Upper Limit Importance
Liver is in range 12 24 3
Fat is in range 15 18 3
Meat is in range 18 33 3
Hardness maximize 1452.75 3285.83 5
Cohesiveness maximize 0.16 0.37 5
Chewiness maximize 244.20 889.28 5

Determination of the results was based by desirability value. As value

approaching 1 indicated an expected result. The result on Table 7 showed that
formula with meat level of 33%, liver level of 12%, and fat level of 15% had the
highest desirability value (0.809), and the second formula with level of meat
25.21%, liver 18.00%, and fat 16,79% had 0.451 on desirability. The optimal

24
formula on textural properties is attributed to sample 8 on the analysis. By these
results, formula 8 could be used as meat ratio for part 2 and 3.

Table 7. Solutions with desirability value from DX software

No Liver Fat Meat Desirability
1 12 15 33 0.809
2 18 16.787 25.213 0.451

4.2. Part 2: Effect of Different Endpoint Temperature on Chicken Liver

Sausage
Table 8 shows the texture profile of chicken liver sausage cooked with
different temperatures (65oC, 75 oC, 85 oC). Compared to the commercial product
of chicken sausage, the samples had a significant (p<0.05) difference on hardness,
cohesiveness, and chewiness, properties. This suggests us that those properties of
sample product mentioned above required to be improved. On the samples, product
cooked to 85oC on the internal temperature had the highest value on hardness, and
chewiness value. The increases of textural properties correlated with continuous
reduction in the amount of extractable proteins. The decreases in the amount of
soluble protein indicates that they were taken up into the gel structure. Further
heating to 70oC above resulted in denser protein matrix (Barbut et al, 1996). The
increases of textural properties seemed to agree with the results of Lin and Mei
(2000), which reported that meat batter containing carrageenan also had the highest
value of hardness and chewiness at the higher endpoint temperatures (76.7oC and
82.2oC).
Among the texture parameters which were tested, hardness is the most
important parameter to consumers at it is determines the commercial value of meat
products (Nurul et al, 2010). On hardness, there were no significant difference
between sample cooked to 65oC and 75oC temperature. Product cooked to 85oC
shows higher value than other cooked products (2.43 ± 0.52). The result of
ANOVA shows that the temperature range didn’t significantly affect the
springiness value. On cohesiveness, commercial product shows significantly
higher values than the cooked samples. Samples cooked to 85oC had the highest
value on cohesiveness in the cooked samples. This could be mean that the internal

25
bonds of product had strengthened by the increases of temperature. Lastly on
chewiness, samples cooked to 85oC had significant difference with samples
cooked to 65oC and 75oC. From the results, author suggested that chicken liver
sausage should be cooked to 85oC to reach the highest value of hardness,
cohesiveness, chewiness properties. According to Varnam and Sutherland (1995),
sausages should be cooked to a core temperature of 85oC. Compared to the
commercial, improvement of textural properties on chicken liver sausage also
suggested due to the poor value in hardness and chewiness properties.

Table 8. Texture profile analysis of chicken liver sausage on different endpoint

temperature compared to the commercial
Product Hardness (kg) Springiness Cohesiveness Chewiness
Comm. 3.82 ± 0.36a 0.96 ± 0.02 0.57 ± 0.05b 2101.75 ± 332.27
o c c
65 C 1.18 ± 0.41 0.78 ± 0.18 0.17 ± 0.05 237.07 ± 21.83b
75oC 1.22 ± 0.37c 0.83 ± 0.07 0.27 ± 0.05c 221.12 ± 48.82b
85oC 2.43 ± 0.52b 0.87 ± 0.14 0.25 ± 0.03a 421.62 ± 83.70a
a-c
Means in a same column with different letters are significantly different (P<0.05)

The experimental data of all color parameters L*, a*, and b* at different
endpoint temperatures are shown in the Table 9. L* value indicates a lighter color,
which is desirable in order to ensure that meat products will have high consumer
acceptance (Resurreccion, 2002 in Furnols & Guerrero, 2014). Samples cooked to
endpoint temperature 85oC showed higher L* value than lower endpoint
temperatures. This could be related to the lightening of meat with increased heat
temperature described by Young and West (2001). The lightening is due to an
increased reflection of light, arising from light scattering by denatured proteins.
The redness (a*) increased significantly on samples cooked to 75oC and reduced
on samples cooked to 85oC. This could due to the denaturation of myoglobin in
meat batter when heated to 85oC. Described by Varnam and Sutherland (1995),
myoglobin in pure solution is denatured when heated to ca. 85oC.
Table 9. Color values of chicken liver sausage on different endpoint temperature
a-b
Temp L a* b*
65oC 74.39 ± 0.59b 3.79 ± 0.04c 22.84 ± 0.15a
75oC 73.06 ± 0.65c 4.08 ± 0.01a 22.89 ± 0.10a
o
85 C 77.06 ± 0.01a 3.96 ± 0.01b 21.91 ± 0.02b
Means in a same column with different letters are significantly different (P<0.05)

26
 As can be seen on Table 10, expressible moisture of samples is significantly
different with commercial product which has the lowest value. This could due to
that commercial sausage used various types of texturizer which could bind water
under application of force such as fiber and different source of proteins. It appeared
that expressible moisture value was not affected by increasing the endpoint
temperature from 65oC – 85oC on chicken-liver sausage. The difference between
samples and commercial products showed that sausage was still lack of water
binding properties which is one of the most important properties of emulsion-type
sausage (Peng, Xing-lian, & Guang-hong, 2009).
Table 10. Expressible moisture of chicken liver sausage on different endpoint
temperature compared to the commercial
Product Expressible Moisture (%)
Commercial 10.77 ± 1.87b
o
65 C 42.79 ± 3.82a
75oC 45.02 ± 1.44a
85oC 42.00 ± 6.96a
a-b
Means in a same column with different letters are significantly different
(P<0.05)

4.3. Part 3: Determining the Optimal Ratio of Different Hydrocolloids

Added to Chicken Liver Sausage
4.3.1. Texture Profile Analysis (TPA)
Texture profile of sausage with different hydrocolloids added such as
Carrageenan (Cr), Alginate (A) and Tapioca starch (T) were analyzed. TPA data
shown in Figure 9 of each variables (hardness, springiness, cohesiveness,
chewiness). The results indicated that sample 6 (4.5% Cr) had the highest hardness,
and chewiness (p<0.05). From the result of sample 5, 8, and 10, the reduction of
Iota carrageenan shows lower value on hardness and chewiness. Sample 7 (4.5% T)
had the lowest value on hardness and springiness. On springiness, sample 2 (4.5%
A) and sample 8 (2.25% A, 2.25% Cr) had the highest value. Lastly on
cohesiveness, sample 5 (2.25% T, 2.25% Cr), sample 6 and sample 8 had the highest
value.
On hardness, model terms suggested by the software are special cubic model
order. Variable A stands for Iota-carrageenan, B for sodium alginate, and C for
tapioca starch. By response, equation acquired as follows:
Y (hardness) = +4375.44 A + 2929.74 B + 2170.33 C − 745.49 AB + 257.80 AC +
2064.89 BC − 16513.77 ABC (4.1)

27
From equation 4.1, it could be seen that there was correlation between one
component against each other affecting to the variable. On springiness, chewiness,
and cohesiveness, model terms suggested are linear model terms as follows:

Y (springiness) = +0.55A + 0.69B + 0.24C (4.2)

Y (chewiness) = +1097.52A + 450.86B + 59.19C (4.3)
Y (cohesiveness) = +0.51A + 0.22B + 0.25C (4.4)
These models were indicating that there are no correlation between component
affecting to the variables. Components such as hydrocolloids added affected
separately to springiness, chewiness, and cohesiveness properties.
1.000 a
5 a
b 0.800 b
Springiness
bcd c
bc cd cd c
Hardness (kg)

cde bc
4 def def def
0.600 de e
efg fg f
3 g
0.400 g
2
0.200
1
0 0.000
1 2 3 4 5 6 7 8 9 10 11 1 2 3 4 5 6 7 8 9 10 11
Formula Formula

0.8 2
a a
Cohesiveness

Chewiness

0.6 bc ab
a 1.5 a
bc cd cd cd 1 b
0.4
d d d c cd bc cde de
cdcde
0.2 0.5 e
0.0
0
1 2 3 4 5 6 7 8 9 10 11 1 2 3 4 5 6 7 8 9 10 11
Formula Formula

Figure 9. Texture profile analysis of chicken-liver sausage. Each value is presented as

mean ± standard deviation. Means above each bar with different letters differ
significantly (P<0.05)

Contour graph was plotted by DX software for each variable. Ranges of

color showed were red, green until blue, which red color showed higher values of
variables and blue showed lower values. From contour graph predicted by DX
software on Figure 10, the addition of Iota carrageenan increasing hardness,
cohesiveness, and chewiness value. This result could be related to study of Ayadi
et al (2009), who reported that addition of Iota carrageenan caused an increase in
hardness and cohesiveness. This could be due to the forming of Ca2+ ion between

28
two sulfate groups of two different double helixes of carrageenan, as followed by
forming inter-macromolecular bonds results in the breaking force of the gel.
Different findings for the effects of carrageenan on texture of comminuted
meat products have been reported. It had been reported by Lin and Mei (2000),
cooked meat batter with Iota-carrageenan had higher in hardness than batter with
alginate. Moreover, the addition of carrageenan increased gel strength of salt
soluble meat protein gels in model systems and hardness of low-fat batter when
increasing carrageenan (Foegeding & Ramsey, 1987 in Nicomrat et al, 2016). From
the Figure 10, it showed that the addition of alginate has increased the springiness
value. This result was in agreement with Santana et al (2013) who found that the
addition of alginate has improved the springiness of sausages made from surimi
powder. This could due to that alginate changed physical state of protein and it
affected the texture of meat products including springiness, and also the ability to
form hydrogen-binding with water (Ensor et al, 1991 in Sarteshnizi et al, 2015).

Figure 10 Contour graphs of TPA results

4.3.2. Color Analysis (Hunter LAB)

Color analysis results are shown on Table 11. Samples 6 (4.5% Cr) were
darker than other sample but had the highest value on redness (a*) and lowest value
on yellowness (b*). This could be related to the result of Koutsopoulos et al
(2008), who reported that sausage with 1-3% carrageenan was darker than highest

29
 Table 11. Color value from chicken-liver sausage
Color value
Formula
L* a* b*
b h
1 74.27 ± 0.14 1.33 ± 0.02 21.12 ± 0.02d
c e
2 73.43 ± 0.42 2.65 ± 0.03 22.05 ± 0.06c
3 73.45 ± 0.67c 2.35 ± 0.04f 21.14 ± 0.21d
4 73.43 ± 0.13a 2.65 ± 0.01d 22.05 ± 0.06g
5 75.19 ± 0.09d 4.50 ± 0.01c 20.53 ± 0.01i
6 69.32 ± 0.15g 6.70 ± 0.01a 18.54 ± 0.02j
7 70.69 ± 0.02f 1.19 ± 0.01j 20.99 ± 0.03e
8 70.46 ± 0.12f 3.11 ± 0.01d 23.19 ± 0.05d
9 70.50 ± 0.08f 1.68 ± 0.01g 22.65 ± 0.04b
e b
10 71.15 ± 0.16 5.95 ± 0.02 20.30 ± 0.04h
11 71.16 ± 0.13e 2.37 ± 0.02f 23.13 ± 0.04a
a-j
Means in a same column with different letters are significantly different (P<0.05)

sausage without carrageenan. Sample 5 (2.25% Cr, 2.25% T) had the highest on
lightness (L*) value. On lightness, model terms which suggested are quadratic
model. The results are no significance impact to the lightness by the formula
proposed. On redness and yellowness, linear model terms were suggested and
equation acquired as follows:
Y (redness) = +5.98A + 2.35B + 0.90C (4.5)
Y (yellowness) = +19.66A + 23.23B + 21.65C (4.6)
that there were no correlation between component affecting to the variables.
Components such as hydrocolloids added affected separately to springiness,
chewiness, and cohesiveness properties.

Figure 11. Model graph of color analysis (L*, a*, b*) results

From redness, the increased level of carrageenan would increase the

redness value. This could be associated to that carrageenan is used to improve
water-binding and emulsion stability on low-fat products which had redder and

30
darker color (Trius and Sebanek, 1996 in Nicomrat et al, 2016). Lastly from
yellowness, samples added with alginate could highly increase the value while the
addition of carrageenan reduce the value. This could due to sodium alginate which
had yellowish color.

4.3.3. Expressible Moisture and Cooking Loss

Table 12 shows the water binding properties of chicken-liver sausage.
Samples added with alginate produced significantly lower cooking loss value than
samples without alginate. As explained by Xiong et al in Nicomrat et al (2016),
alginate reduced cooking loss on pork sausage. It was associated to formation of
hydrogen binding with water. Alginate also reported to its ability to form a more
heat-stable alginate gel, resulted a higher water holding capacity at higher cooking
temperatures (Lin & Mei, 2000). Moreover, it was explained that the use of this
gums resulted in softer, crumbly, and slippery texture which could be seen on
texture analysis results. So, the addition of alginate was undesirable to low-fat (4%)
products. As could be seen on the Figure 12, increased level of starch would
produce higher cooking loss value. On the contrary, increased level of alginate
would had lower cooking loss value. Carrageenan could also be used to reduce
cooking loss in a lower level than alginate.
Table 12. Cooking loss and expressible moisture of chicken-liver sausage
Formula Cooking Loss (%) Expressible Moisture (%)
a
1 1.35 ± 0.06 37.56 ± 2.21bc
2 0.36 ± 0.02efg 37.65 ± 0.25bc
3 0.42 ± 0.14def 32.66 ± 2.99bcd
4 0.46 ± 0.05de 31.02 ± 3.96bcd
5 0.74 ± 0.03c 31.77 ± 5.32bcd
6 0.57 ± 0.07d 26.01 ± 3.30d
7 1.05 ± 0.03b 47.58 ± 0.64a
8 0.22 ± 0.04gh 25.71 ± 1.08d
9 1.21 ± 0.09a 38.56 ± 3.57b
fgh
10 0.27 ± 0.07 31.05 ± 2.04bcd
11 0.19 ± 0.05h 30.45 ± 4.75cd
a-e
Means in a same column with different letters are significantly different (P<0.05)

On cooking loss, model terms suggested are linear and equation acquired as
follows: Y (cooking loss) = +0.53A + 0.075B + 1.20C (4.7)

31
The linear model indicated that there were no correlation between component
affecting to the variables. Components such as hydrocolloids added affected
separately to springiness, chewiness, and cohesiveness properties. Special cubic
model were suggested for expressible moisture and defines as:
𝑌 (𝐸𝑀) = + 25.55𝐴 + 31.43𝐵 + 43.91𝐶 (4.8)
Sample 6 and 8 had the lowest expressible moisture while sample 7 (4.5%
T) had the highest value. It was explained by Sarteshnizi et al (2015) that starch
increased expressible moisture and decrease cohesiveness, probably due by to
difference in temperature of starch granule gelatinization and meat protein
denaturation. Iota-carrageenan solubilizes at 50oC and able to penetrate the matrix
before the myofibrillar protein gel has set (Trius & Sebranek, 1996). By model
graph on Figure 12, it was shown that increased level of carrageenan could reduce
expressible moisture value. Furthermore, alginate could also reduce expressible
moisture in lower level than carrageenan. This may be due to the functional groups
of each hydrocolloid for water-binding. Iota-carrageenan had two sulfate groups
which could improve moisture retention in meat product (Yugushi et al, 2003 in
Nicomrat et al, 2016). Alginate had a lot of carboxyl groups in its structure that an
bind water and promote strong electrostatic repulsion between the chain (Sanchez
et al, 1995 in Nicomrat et al, 2016).

Figure 12. Contour graphs of expressible moisture and cooking loss

32
4.3.4. Appearance of Products
From the appearance of products in Figure 13, it was showed that samples
without added alginate had smooth surface without any dark spots. This
characteristic appeared on sample 1, 5, 6 and 7. Dark spots as could be seen are
the results of the addition of alginate which had cold set binder ability. It can form
a gel at room temperature from the reaction between alginate salt and a calcium
ion (Suklim et al, 2004 in Nicomrat et al, 2016). Therefore, the dark spots may due
to the gelling of alginate before batter stuffing into the casing then got darker color
after cooking.

1 2 3 4 5 6

7 8 9 10 11

Figure 13. Appearance of Chicken-liver sausage samples

4.3.5. Sensory Evaluation

According to Nurul et al (2010), determination of good textural in meat
products should be done together with sensory test to find the suitable range
preferred by consumers. Therefore, samples 1 (Control), 5, 6, 8, and 10 has chosen
by appearance and higher textural properties value to be included in sensory test.
On color, no different were found among samples (p<0.05). The results on Figure
14 explained that the range of value (L*, a*, b*) in color analysis was not correlated
to the preference by consumers. Sample 6 had the highest score among samples in
appearance may due to the smooth surface and had more redness (a*) in color.

33
 Color Appearance
10 10
a

9-point hedonic scales

9-point hedonic scales
b b b
8 8 c
6 6

4 4

2 2

0 0
1 5 6 8 10 1 5 6 8 10
Sample No. Sample No.

Texture Overall
9 9 a
a b

9-point hedonic scales

8
9-point hedonic scales

8 b bc c c
c c 7
7 c
6 6
5 5
4 4
3 3
2 2
1 1
0 0
1 5 6 8 10 1 5 6 8 10
Sample No. Sample No.

Figure 14. Sensory scores of selected samples. Each value is presented as mean ±
standard deviation. Means above each bar with different letters differ significantly
(P<0.05)

Sample 8 had the lowest score in appearance could due to the increased level
of alginate which resulted in small dark spots scattered in the surface. On texture,
sample 6 had the highest score among samples. This could due to it had the highest
value in hardness which mentioned by Nurul et al (2010) as the most important
parameter to consumers as it determines the commercial value of meat products.
According to Dingstad et al in Nurul et al, 2010, frankfurters with hardness of 4.73
kg and above will have at least 60% consumers willing to buy it. Sample 6 had the
highest score of hardness (4.32 kg), which had the lowest difference with the
referred value (4.73 kg). Moreover, Caceres et al in Nurul et al, 2010 found that
gumminess and chewiness behave similarly to hardness, which sample 6 had also
the highest score in chewiness, followed by sample 10. On overall acceptability,
sample 6 had the highest score among other samples. This result showed that sample
6 which had the highest value in parameters such as hardness, chewiness, redness

34
could be correlated to the highest preference by consumers. By Pearson correlation
test, hardness had the strongest positive correlation (0.963) with texture parameter
on sensory evaluation.
4.3.6. Formula optimization
Significant variables related such as hardness, chewiness, cohesiveness,
springiness, redness (a*), cooking loss, and expressible moisture were used as
criteria of optimization process showed at Table 13.

Table 13. Criteria of optimization on ratio of hydrocolloids

Criteria Goal Lower Limit Upper Limit Importance
Hardness (g) maximize 2192.56 4324.01 3
Cohesiveness maximize 0.21 0.51 3
Springiness maximize 0.18 0.72 3
Chewiness (g) maximize 107.26 1189.71 3
a* maximize 1.19 6.70 3
Cooking loss (%) minimize 0.19 1.35 3
Expressible Moisture (%) minimize 25.71 47.58 3

The result on Table 14. showed that formula with level of carrageenan 4.5%,
alginate 0%, tapioca starch 0 % had the highest desirability value (0.861), and the
second formula with level of carrageenan 2.25%, alginate 2.25 %, tapioca starch 0
% had 0.684 on desirability.

Table 14. Solutions with desirability value from DX software (%)

No Iota-Crg S Alginate Tpc Starch Desirability
1 4.5 0 0 0.861
2 2.25 2.25 0 0.684

35
 CHAPTER V
CONCLUSION & SUGGESTION
5.1 Conclusion
On this study, variances of chicken liver, meat, and fat ratio and different
hydrocolloids ratio were proposed to produce chicken liver sausages with higher
textural properties. Different endpoint temperatures (65oC, 75oC, 85oC) on chicken
liver sausages also examined to determine the effect of endpoint temperature on
cooking products. Formulas were designed by DX 7.0 software using Mixture
Design feature. Formulation of chicken liver, meat and fat had significant impact
on hardness, chewiness, and cohesiveness. Formula optimization was conducted
and resulted that sausage with meat level of 33%, liver level of 12%, and fat level
of 15% had the highest desirability value (0.809).
By the results, cooking chicken liver sausages to 85oC endpoint temperature
would produce product with higher hardness, cohesiveness, and chewiness
(p<0.05). Samples cooked to endpoint temperature 85oC showed higher L* value
than lower endpoint temperatures. The redness (a*) increased significantly on
samples cooked to 75oC and reduced on samples cooked to 85oC.
Hydrocolloids such as Iota carrageenan (Cr), sodium alginate (A) and
tapioca starch (T) were added by Simplex-Centroid Design to improve the textural
properties of chicken liver sausages. The model system had a significant impact on
hardness, springiness, chewiness, cohesiveness, color values (a* and b*), cooking
loss, and expressible moisture. Addition of Iota-carrageenan increases hardness,
chewiness, cohesiveness, and redness (a*), while addition of alginate increases
springiness and yellowness (b*) of the products. Samples 1,5,6,8, and 10 were used
in sensory evaluation and resulted that sample 6 had the highest score in texture,
appearance, and overall parameters (p<0.05). Formula optimization was resulted
that sausage with Iota-carrageenan level 4.5%, alginate 0%, and starch 0% had the
highest desirability value (0.846).

36
5.2 Suggestion
For future works, variances of texturizer agent such as fiber, phosphates,
and various sources of protein could be studied to improve textural properties of
chicken liver sausages. Pre-mix ingredients should also be added to products to
improve taste preference by consumers. Following study should focused on the
effect of chicken liver to the quality characteristics on the products.
As gelation curve could be resulted and showed a characteristic peak during
heating of sausage batter, author suggested that thermo-rheological properties
measurements should be conducted. Advanced studies of textural properties such
as microstructure also could be proposed using Scanning Electron Microscopy
(SEM).

37
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41
 APPENDIX

Appendix 1. Springiness value graph on chicken liver sausages with different ratio of
liver,meat and fat. Each value is presented as mean ± standard deviation. Means
above each bar with different letters differ significantly (P<0.05)

1.2

1 abc ab abc a
abc abc abc abc
abc abc
bc abc
0.8 c
Springiness

0.6

0.4

0.2

0
1 2 3 4 5 6 7 8 9 10 11 12 13
Formula

Appendix 2. ANOVA statistical result of expressible moisture percentage on part 1

Sum of p-value
Source Squares df Mean Square F Value Prob > F
not
Model 27.69 2 13.84 0.56 0.5891 significant
Linear Mixture 27.69 2 13.84 0.56 0.5891
Residual 247.99 10 24.80
not
Lack of Fit 63.21 7 9.03 0.15 0.9826 significant
Pure Error 184.78 3 61.59
Cor Total 275.68 12

42
Appendix 3. ANOVA statistical result of hardness value on part 1

Sum of Mean p-value Prob >

Source Squares df Square F Value F
Model 2346152.79 6.00 391025.46 5.08 0.0343 significant
Linear
Mixture 165474.83 2.00 82737.42 1.07 0.3991
AC 724441.59 1.00 724441.59 9.41 0.0220
AB(A-B) 1281729.87 1.00 1281729.87 16.65 0.0065
AC(A-C) 1031656.07 1.00 1031656.07 13.40 0.0106
BC(B-C) 64849.10 1.00 64849.10 0.84 0.3942
Residual 461977.37 6.00 76996.23
not
Lack of Fit 113858.06 3.00 37952.69 0.33 0.8084 significant
Pure Error 348119.31 3.00 116039.77
Cor Total 2808130.16 12.00

Appendix 4. ANOVA statistical result of springiness value on part 1

Sum of Mean p-value Prob >

Source Squares df Square F Value F
not
Model 0.01 2.00 0.01 3.04 0.0927 significant
Linear
Mixture 0.01 2.00 0.01 3.04 0.0927
Residual 0.02 10.00 0.00
not
Lack of Fit 0.01 7.00 0.00 0.45 0.8260 significant
Pure Error 0.01 3.00 0.00
Cor Total 0.03 12.00

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Appendix 5. ANOVA statistical result of cohesiveness value on part 1

Sum of Mean p-value Prob >

Source Squares df Square F Value F
Model 0.04 5.00 0.01 4.88 0.0305 significant
Linear
Mixture 0.02 2.00 0.01 6.32 0.0270
AB 0.02 1.00 0.02 11.73 0.0111
AC 0.02 1.00 0.02 8.62 0.0218
ABC 0.01 1.00 0.01 7.91 0.0260
Residual 0.01 7.00 0.00
not
Lack of Fit 0.00 4.00 0.00 0.47 0.7631 significant
Pure Error 0.01 3.00 0.00
Cor Total 0.06 12.00

Appendix 6. ANOVA statistical result of chewiness value on part 1

Sum of Mean p-value Prob >

Source Squares df Square F Value F
Model 209790.58 2.00 104895.29 4.99 0.0314 significant
Linear
Mixture 209790.58 2.00 104895.29 4.99 0.0314
Residual 210307.56 10.00 21030.76
not
Lack of Fit 183118.78 7.00 26159.83 2.89 0.2069 significant
Pure Error 27188.78 3.00 9062.93
Cor Total 420098.14 12.00

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Appendix 7. ANOVA statistical result of hardness value on part 3
Sum of Mean
Source Squares df Square F Value p-value Prob > F
Model 3400294.08 6.00 566715.68 21.14 0.0054 significant
Linear
Mixture 2873058.77 2.00 1436529.38 53.58 0.0013
AB 23476.76 1.00 23476.76 0.88 0.4024
AC 4097.33 1.00 4097.33 0.15 0.7158
BC 180131.69 1.00 180131.69 6.72 0.0606
ABC 292483.60 1.00 292483.60 10.91 0.0299
Residual 107248.84 4.00 26812.21
Cor Total 3507542.92 10.00

Appendix 8. ANOVA statistical result of springiness value on part 3

Mean F p-value
Source Sum of Squares df Square Value Prob > F
Model 0.16 2.00 0.08 15.46 0.0018 significant
Linear
Mixture 0.16 2.00 0.08 15.46 0.0018
Residual 0.04 8.00 0.01
Cor
Total 0.21 10.00

Appendix 9. ANOVA statistical result of cohesiveness value on part 3

Sum of Mean F p-value
Source Squares df Square Value Prob > F
Model 0.08 2 0.04 7.00 0.0175 significant
Linear
Mixture 0.08 2 0.04 7.00 0.0175
Residual 0.05 8 0.01
Cor Total 0.12 10

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Appendix 10. ANOVA statistical result of chewiness value on part 3
Sum of Mean F p-value Prob
Source Squares df Square Value >F
Model 833986.77 2 416993.39 11.53 0.0044 significant
Linear
Mixture 833986.77 2 416993.39 11.53 0.0044
Residual 289436.96 8 36179.62
Cor
Total 1123423.73 10

Appendix 11. ANOVA statistical result of lightness value on part 3

Sum of Mean F
Source Squares df Square Value p-value Prob > F
Model 26.94 5 5.39 2.85 0.1376 not significant
Linear
Mixture 2.04 2 1.02 0.54 0.6136
AB 1.92 1 1.92 1.01 0.3604
AC 21.40 1 21.40 11.32 0.0200
BC 0.19 1 0.19 0.10 0.7629
Residual 9.46 5 1.89
Cor
Total 36.40 10

Appendix 12. ANOVA statistical result of redness value on part 3

Sum of Mean p-value Prob
Source Squares df Square F Value >F
Model 20.88 2 10.44 6.71 0.0195 significant
Linear
Mixture 20.88 2 10.44 6.71 0.0195
Residual 12.45 8 1.56
Cor Total 33.33 10

Appendix 13. ANOVA statistical result of yellowness value on part 3

Sum of Mean F p-value Prob
Source Squares df Square Value >F
Model 10.46 2 5.23 4.89 0.0410 significant
Linear
Mixture 10.46 2 5.23 4.89 0.0410
Residual 8.56 8 1.07
Cor Total 19.02 10

46
Appendix 14. ANOVA statistical result of cooking loss percentage on part 3
Sum of Mean F p-value
Source Squares df Square Value Prob > F
Model 1.01 2 0.51 6.03 0.0253 significant
Linear
Mixture 1.01 2 0.51 6.03 0.0253
Residual 0.67 8 0.08
1
Cor Total 1.69 0

Appendix 15. ANOVA statistical result of expressible moisture percentage on part 3

Sum of Mean F p-value Prob >
Source Squares df Square Value F
significa
Model 263.71 2 131.86 7.79 0.0132 nt
Linear
Mixture 263.71 2 131.86 7.79 0.0132
Residual 135.40 8 16.93
Cor Total 399.11 10

47