MAKERERE UNIVERSITY

COLLEGE OF AGRICULTURE AND ENVIRONMENTAL SCIENCES

DEPARTMENT OF AGRICULTURAL AND BIOSYSTEMS
ENGINEERING

OPTIMIZING THE PROCESSING CONDITIONS

AND FORMULATION OF A BANANA-
VEGETABLES INSTANT SOUP FLOUR

PADDY AINEBYONA
BSc. Agric. Eng. (Mak)

A THESIS SUBMITTED TO THE DIRECTORATE OF RESEARCH AND

GRADUATE TRAINING IN PARTIAL FULFILLMENT FOR THE
AWARD OF MASTER OF SCIENCE DEGREE IN AGRICULTURAL
ENGINEERING OF MAKERERE UNIVERSITY

FIRST DRAFT, August 2019

DECLARATION

I, Paddy Ainebyona, declare that this research thesis is my original work and has never been
submitted to any University or institution of higher learning for an academic award.
………………………………………………………………..…………………………
Paddy Ainebyona Date

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APPROVAL

This thesis has been submitted with the endorsement of the following supervisors:

Signature................................................. Date........................................................
Dr. Julia Kigozi
Department of Agricultural and Biosystems Engineering
School of Food Technology, Nutrition, and Bioengineering

Signature................................................. Date.........................................................
Dr. Ivan MuziraMukisa
Department of Food Technology and Nutrition
School of Food Technology, Nutrition, and Bioengineering

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COPYRIGHT

© Copyright is reserved by author

iii
DEDICATION

This special project report is dedicated to my parents Mr. Kamushwa George, Mrs. Birungi
Lydia for their positive motivation and to their grandson Abdul AwwalAinebyona.

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ACKNOWLEDGEMENT

I thank the Almighty Allah,the most gracious, the most merciful for the gift of life, knowledge,
creativity, understanding, courage and strength that He has blessed me with while pursuing my
graduate studies at this noble university. There is no doubt; he is all-knowing and the master
Planner for mankind.

I sincerely extend my heartfelt appreciation to my supervisors Dr. Julia Kigoziand Associate

Professor, Ivan MuziraMukisafor the guidance during data collection, and writing this special
thesis. Dr. Julia participated in the initial identification stages that got me the funding for this
special project report.Thank you very much for believing in me and shaping me to become what
I have become today. May the Almighty reward and bless your more and more.

I gratefully extend my sincere appreciation to Dr. Susan Balabba of the school of Forestry and
natural resources conservation who also was the principal investigator for this project. Dr Susan
managed the finances of the project. Thank you so much for all the great support that has seen
me complete this research in time. May the Almighty bless you more abundantly.

To everyone who contributed directly or indirectly to the success of this special project and final
special thesis especially my course matesBasika Elisa, Mutonyi Cynthia Wandeka, and Oshaba
Brenda not forgetting DeoBusuulwa who spent most of the time in laboratory data collection at
SFTNB. Thank you very much for your support. May the Almighty reward you with lots of
wisdom to see success in this world.

My appreciation to Mrs. FildaAya can‟t be under looked. She has opened doors for me whenever
I seek for assistance especially in the software part as well as in sample preparations that saw me
perfect the product. Thank you very much for your kind support.

Lastly I am very much grateful to RELOAD, a German based organization for funding all my
graduate studies at the most prestigious university in Uganda, Makerere University. Thank you
very much RELOAD for considering me as one of your beneficiaries.

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TABLE OF CONTENTS
DECLARATION............................................................................................................................ i
APPROVAL .................................................................................................................................. ii
COPYRIGHT ............................................................................................................................... iii
DEDICATION.............................................................................................................................. iv
ACKNOWLEDGEMENT ............................................................................................................ v
LIST OF FIGURES ..................................................................................................................... xi
LIST OF TABLES ...................................................................................................................... xii
LIST OF ABBREVIATIONS ................................................................................................... xiii
CHAPTER ONE: INTRODUCTION ......................................................................................... 1
1.1. Background ............................................................................................................................. 1
1.2. Problem statement ................................................................................................................... 2
1.3. Objectives ................................................................................................................................ 3
1.3.1. Main objective .............................................................................................................. 3
1.3.2. Specific objectives ........................................................................................................ 3
1.4. Hypotheses .............................................................................................................................. 4
1.5 Justification of the study.......................................................................................................... 4
CHAPTER TWO: LITERATURE REVIEW ............................................................................ 5
2.1. Bananas ................................................................................................................................... 5
2.1.1. Banana production in Uganda ....................................................................................... 5
2.1.2. Physical characteristics of bananas ............................................................................... 6
2.1.3. Chemical characteristics of bananas ............................................................................. 6
2.1.4. Nutritional value of banana ........................................................................................... 7
2.1.5. Banana value chain in Uganda ...................................................................................... 8
2.1.6. Processing of bananas into value added products ......................................................... 8
2.2. Description of soups .............................................................................................................. 10
2.2.1. Commercial soups ....................................................................................................... 10
2.2.1.1. Canned .................................................................................................................. 10
2.2.1.2. Dried ..................................................................................................................... 11
2.3. Choice of supplements to banana flour. ................................................................................ 11
2.3.1. Grain Amaranth .......................................................................................................... 11
2.3.2. Pumpkin ...................................................................................................................... 12
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 2.3.3. Carrot .......................................................................................................................... 12
2.3.4. Mushrooms ................................................................................................................. 13
2.3.5. Tomatoes ..................................................................................................................... 13
2.4. Process optimization.............................................................................................................. 14
2.4.1. Background ................................................................................................................. 14
2.4.2. Optimization methods ................................................................................................. 15
2.4.3. Choice of an optimization method .............................................................................. 15
2.4.4. Experimental designs in RSM .................................................................................... 16
2.4.5. Steps involved in the application of response surface methodology .......................... 16
2.4.6. Determination of optimum conditions for multiple variable responses in soups. ...... 16
2.5. Extrusion ............................................................................................................................... 17
2.5.1. Types of extruders....................................................................................................... 17
2.5.1.1. Single-screw extruders .......................................................................................... 17
2.5.1.2. Twin screw extruders ............................................................................................ 18
2.5.2. Processing parameters affecting the nutritional quality of extruded products ............ 18
2.5.2.1. Moisture content ................................................................................................... 19
2.5.2.2. Barrel Temperature ............................................................................................... 19
2.5.3. Effect of extrusion cooking on nutritional quality ...................................................... 20
2.5.3.1. Protein retention and digestibility ......................................................................... 20
2.5.3.2. Vitamin A retention .............................................................................................. 20
2.5.3.3. Mineral bioavailability .......................................................................................... 20
2.6. Instant products ..................................................................................................................... 21
2.7. Sensory evaluation ................................................................................................................ 21
2.8. Shelf life studies of food products ......................................................................................... 21
2.8.1. Definition of shelf life ................................................................................................. 22
2.8.2. Factors affecting shelf life .......................................................................................... 22
2.8.2.1. Product characteristics .......................................................................................... 22
2.8.2.2. Distribution environment ...................................................................................... 22
Intrinsic factors ............................................................................................................... 23
Extrinsic factors.............................................................................................................. 23
2.8.3. Predicting shelf-life/shelf life testing .......................................................................... 23
2.8.4. Selection criteria to assess shelf life ........................................................................... 24
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 2.8.5. Types of shelf life testing ............................................................................................ 24
2.8.5.1. Direct method: ...................................................................................................... 24
2.8.5.2. Accelerated shelf life studies ................................................................................ 25
2.8.6. Measurement of product shelf-life .............................................................................. 27
2.8.6.1. Sensory panels ...................................................................................................... 27
2.8.6.2. Physical measurements ......................................................................................... 28
2.8.6.3. Chemical measurements ....................................................................................... 28
2.8.6.4. Microbiological stability determinations. ............................................................. 28
CHAPTER THREE: RESEARCH MANUSCRIPT ONE ...................................................... 30
Optimization of the effect of formulation on nutritional quality, texture and
physicochemical properties of banana-vegetable soup powder .............................................. 30
3.1. Introduction ........................................................................................................................... 30
3.2. Methods and materials........................................................................................................... 31
3.2.1. Raw material selection for use in this study ............................................................... 31
3.2.2. Sample preparation ..................................................................................................... 31
3.2.3. Determining the optimum incorporation level of bananas and selected vegetables for
use in the formulation of the banana-vegetable soup ............................................................ 32
3.2.4. Proximate composition of the soup samples ............................................................... 33
3.2.4.1. Determination of moisture content ....................................................................... 33
3.2.4.2. Determination of crude protein content ................................................................ 33
3.2.4.3. Determination of fat content ................................................................................. 34
3.2.4.4. Determination of Ash content ............................................................................... 34
3.2.4.5. Dietary fiber content ............................................................................................. 34
3.2.4.6. Determination of total carbohydrates content ....................................................... 35
3.2.4.7. In vitro protein digestibility .................................................................................. 35
3.2.4.8. Determination of gross energy content ................................................................. 36
3.2.5. Physicochemical properties of the flours .................................................................... 36
3.2.5.1. Bulk density .......................................................................................................... 36
3.2.5.2. Water absorption capacity..................................................................................... 37
3.2.5.3. Water absorption index and water solubility index .............................................. 37
3.2.6. Pasting properties of soups ......................................................................................... 37
3.2.7. Sensory evaluation of formulations for acceptability of the formulations. ................ 37

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 3.2.8. Data analysis ............................................................................................................... 38
3.3. Results ................................................................................................................................... 38
3.3.1. Effect of formulations on the Nutritional composition of the soups .......................... 38
3.3.2. Effect of formulations on micro nutrients composition of the soups.......................... 41
3.3.3. Effect of formulations on physicochemical properties of the soups ........................... 43
3.3.4. Effect of formulations on pasting properties of the soups .......................................... 45
3.4. Optimization of responses ..................................................................................................... 48
3.4.1. Fitting the response surface models for Nutritional composition of the enriched
banana vegetable soup. .......................................................................................................... 48
3.4.1.1. Model graphs ........................................................................................................ 49
3.4.1.2. Predictive models for nutrition composition of the formulations ......................... 50
3.4.1.3. Optimal solutions for selection of nutrient enriched soup flour ........................... 53
3.5. Discussion and Conclusion ................................................................................................... 54
3.6. References ............................................................................................................................. 56
CHAPTER FOUR: RESEARCH MANUSCRIPT TWO ....................................................... 58
Optimization of extrusion conditions and determination of the effects of incorporation of
cooked vegetable-chicken mixture on sensory acceptability, nutritional composition and
physico-chemical properties of the instant soup powder. ....................................................... 58
4.1. Introduction ........................................................................................................................... 58
4.2. Materials and methods........................................................................................................... 59
4.2.1. Materials ..................................................................................................................... 59
4.2.2. Methods....................................................................................................................... 60
4.2.2.1. Method of preparation........................................................................................... 60
4.2.2.2. The central composite design used to optimize levels of feed moisture content,
screw speed and the barrel temperature of the extruded flours ......................................... 60
4.2.2.3. Reconstitution experiment to determine the most acceptable flours .................... 61
4.2.2.4. Effect of extrusion................................................................................................. 61
4.2.2.5. Preparation of composite flour soups for acceptability screening ........................ 62
4.2.2.6. Sensory acceptability of the composite flour soups. ............................................. 62
4.2.2.7. Nutritional and physicochemical properties ......................................................... 62
4.3. Results ................................................................................................................................... 63
4.3.1. Overall acceptability of the formulation ..................................................................... 64

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 4.3.2. Variations in moisture content, fiber content, fat content, ash content, Vitamin A and
Vitamin C in the formulations. ...............................................Error! Bookmark not defined.
4.4. Optimization of responses ..................................................................................................... 67
4.4.1. Predictive models describing variability of dependent variables in the soups. .......... 67
4.5. Conclusion and recommendations......................................................................................... 70
4.6. References ............................................................................................................................. 70
CHAPTER FIVE: RESEARCH MANUSCRIPT THREE ..................................................... 70
Estimating effect of incorporation of cooked chicken-vegetable mix on shelf-stability of the
instant soup flours using accelerated shelf-life testing. ........................................................... 71
5.1. Introduction ........................................................................................................................... 71
5.2. Materials and methods........................................................................................................... 72
5.2.1. Yeasts and molds ........................................................................................................ 72
5.2.2. Total plate count ......................................................................................................... 72
5.2.3. Total coliforms count .................................................................................................. 73
5.2.3.1. Calculations........................................................................................................... 73
5.3. Results and discussion ........................................................................................................... 74
5.3.1. Microbial load in the flours......................................................................................... 75
5.3.1.1. Total plate count ................................................................................................... 75
5.3.1.2. Total coliform ....................................................................................................... 75
5.3.1.3. Yeasts and Molds .................................................................................................. 75
5.4. Conclusion ............................................................................................................................. 75
References .................................................................................................................................... 76

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LIST OF FIGURES

Figure 2.1: Classification of Bananas (Karamura et al., 1998) ..................................................... 5

Figure 2.2: Banana end markets in Uganda and consumption (Source: Bill and Melinda gates
foundation report, 2009) ................................................................................................................. 8
Figure 2.3: Flow chart showing processing of bananas into different products (Mohapatra et al.,
2010) ............................................................................................................................................... 9
Figure 2.4: A cross-section of a single-screw extrusion-cooker: .................................................. 18
Figure 2.5: Flow chart illustrating steps for calculating the shelf life of food products by direct
method (Source: NZFSA, 2005) ................................................................................................... 25
Figure 3.1: Variation of nutrition values in the formulation;Fat content (FC), Protein content
(PC), Carbohydrates and Starch content (SC). ............................................................................. 39
Figure 3.2: Variation in mineral composition and micronutrients; Iron content, Zinc content,
Potassium content and Vitamin C in the formulations ................................................................. 42
Figure 3.3: Variation of physicochemical properties in the formulation; bulk density (ρb),
Foaming Capacity (FC) and foaming stability (FS) ..................................................................... 44
Figure 3.4: Pasting properties of the soup samples; Peak Viscosity, holding viscosity, final
viscosity, setback viscosity, peak time and pasting temperature. ................................................. 47
Figure3.5: 3-D response surface plots showing the effect of ingredient mixes on Zinc content,
IVPD, Fat and protein Content. .................................................................................................... 49
Figure 4.1: Reconstitution of extruded samples at different conditions ....................................... 61
Figure 4.2: Overall acceptability of the sample formulations....................................................... 64
Figure 4.3: Variations in nutrient composition; moisture content, fiber content, fat content, ash
content, Vitamin A and Vitamin C in the formulations ................................................................ 66
Figure 4.4: Response surface plots showing effect of varying different levels of moisture and
temperature and chicken composition on Vitamin A, Vitamin C, Ash content, Fat, Fiber content
and Overall acceptability. ............................................................................................................. 69
Figure 5.1: Microbial changes in the soup samples during storage .............................................. 74

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LIST OF TABLES

Table 2.1 Chemical composition of matooke from different cultivars ........................................... 7

Table 2.2: The nutritional composition of grain amaranth and vegetables (Value per 100g) ...... 13
Table 2.3: Recommended daily Nutritional requirements for 2-5 years old children .................. 14
Table 3.1: Proportions of bananas and selected vegetables obtained using Nutri-survey ............ 32
Table 3.2: Proximate composition of different flour formulations for making soups .................. 40
Table 3.3. Micronutrients compositions of the samples ............................................................... 41
Table 3.4: Physicochemical properties ......................................................................................... 43
Table 3.5: Pasting properties of flour samples.............................................................................. 47
Table 3.6: Coefficients for the fitted first order and second-order polynomials representing the
relationship between the response and formulations of vegetable chicken soup mixture ............ 48
Table 3.7: Optimal solutions for nutritious and viscous soup formulations. ................................ 54
Table 4.1: Formulations for preparation of chicken vegetable mix .............................................. 59
Table 4.2: Coding of flour vegetable-chicken mixes. ................................................................... 60
Table 4.3: Central Composite Design used to optimize feed moisture, screw speed and barrel
temperature for the production of instant banana-based soup flours ............................................ 61
Table 4.4: Consumer acceptability scores of soups from different flour formulations ................ 63
Table 4.5: Proximate composition of extruded samples ............................................................... 65
Table 4.6: Optimal solutions for nutritious and viscous soup formulations. ................................ 68

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LIST OF ABBREVIATIONS

& And

µg micrograms

AAA-EAHB Musa, spp; East African Highland Bananas

ANOVA Analysis of Variance

AOAC Association of Official Analytical Chemists

ASLT Accelerated Shelf Life Testing

CCD Central Composite Design

DFA Desirability Function Approach

Dr. Doctor

et al., And others

FOSS Free and open-source software

g grams

Ha Hectare

HCl Hydrochloric Acid

IBM International Business Machines

IITA International Institute of Tropical Agriculture

IU International unit

IVPD In Vitro Protein Digestibility

kg Kilogram

MD Mixture Design

NaOH Sodium hydroxide

ND Not Defined

NZFSA New Zealand Food Safety Authority

o
C Degrees Celsius

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RAE Retinol Activity Equivalents

RCCN Research Centers Collaborative Network

RELOAD Reducing Losses and Adding Value

RSM Response Surface Modeling

RTE Ready To Eat

RVA Rapid ViscoAnalyser

SD Standard Deviation

SFTNB School of Food Technology, Nutrition and Bioengineering

UK United Kingdom

USA United States of America

USDA United States Department of Agriculture

WAI Water Absorption Index

WSI Water Solubility Index

xiv
CHAPTER ONE: INTRODUCTION

1.1. Background

Banana fruits are commonly grown in the tropical climates (Daramola & Osanyinlusi, 2006). Global
annual production of bananas stands at 72.5 million metric tons (Radha Krishna et al., 2012). About
eleven million metric tons of these are produced in Uganda (Tumutegyereize et al., 2011; UBOS,
2016). This makes Uganda the second world‟s leading producer of cooking bananas after India which
has an annual capacity of 27.575 million tons (Daily Records, 2018). In Uganda, production of bananas
is done by 70% of farmers (Bagamba et al., 2007) emanating mainly from western and central parts of
the country. The western region contributesabout 68% of total bananas to the market (Daily Records,
2018). Banana business contributes 42%to rural household income (Kawongolo, 2013) and serves as a
staple food for 70% of Ugandans.

Banana production and marketingfaces challenges in developing countriessuch as; delays in

transportation due to poor road networks and transport means, poor storage systems, poor post-harvest
technologies and fluctuating prices and market demands.These factorscontribute to ripening and
senescence thuscausing post-harvest lossesin bananas. Muranga et al., (2010), estimates these losses to
between 22 to 45% of total production translating to annual losses of between $411.9-$842.5 millions.
In an attempt to reduce banana post harvest losses most studies have focused on development of shelf
stable products mainly focussing on drying to obtain flours. Muranga et al.,(2010)developed a raw
Tooke flour for use in confectionaries and has also developed a number of products through
Presidential Innitiative on Banana Industrial Development (PIBID). Most products developed from
bananas have focussed on addressing issues related to malnutrition reported to be one of the rampant
causes of death to children in the country.

Global health reportsattribute malnutrition due to lack of micronutrients asone of the major causes of
health issues in children(Herrador et al., 2014). More than 2 billion people most of whom are children
lack key vitamins and minerals such as Vitamins A;C;B6;B12, Iron, zinc, and potassium (Herrador et
al., 2014) contributing to more than 50% of childhood mortality in developing countries (Thaoge et al.,
2003). In Uganda, low levels of iron has increased anemic rates to 73% (Tidemann-Andersen et al.,
2011; Tumwegamire et al., 2011) while the prevalence of zinc deficiency ranges between 20–69%
among children (Akandeet al., 2017).This indicates that commonly consumed staplesare deficient in

1
protein, vitaminsand nutrients required for normal body health. Development of nutrient dense foods
from availableraw materials could provide means of reducing nutrient deficiencies in diets(Amegovu et
al., 2014). In this study, development of nutrient dense soup from bananas was explored.
Analysis of banana flour shows high starch content, extended shelf life due to low sugar and fat, high
level of potassium and low tannins. Banana flour however, is deficient in key micro nutrients such as
vitamins and minerals like iron and zinc. This therefore implies that there are limitations in
development of products from bananas. There is need to add supplements that can improve banana
nutrient levels to make it a suitable raw material for product development. Banana could be
supplemented with grains, vegetables, legumes to make it possible to develop products such as instant
porridges, soups, among others. In this study options for development of nutrient dense soup for
children below five years of age were explored to reduce reported cases related to nutrient defficiences
in current staple diets. Required nutrients were obtained from grains and vegetables.However, to reduce
the time spent in cooking the soup and to improve on the nutrient concentration of the soup, extrusion
cooking was used(Abdel-haleem and Omran, 2014).

Extrusion being a high temperature-short time process has numerous advantages such as versatility,
high productivity, low operating costs, energy efficiencyand high quality products (Milán-Carrillo et
al., 2012). Extrusion increases digestibility of starch and protein (Diaz et al., 2013). Extrusion also
breaks down mineral-antinutrient complexes through hydrolysis thereby increasing mineral availability
in extrudates, altering vitamins, and eliminating anti-nutrient factors such as phytic acid which modifies
the nutritional properties of the extrudates(Pathaniaet al., 2013; Sundarrajan, 2014).

1.2. Problem statement

Cooking banana is reported to be one of the staple foods in Uganda with the highest (220 kg) per capita
consumption in the world (Kiiza, Abele, & Kalyebara, 2004). Bananas are consumed in various forms,
such as cooked green, cooked ripe, cooked in the peel, steamed, made into juice, ripened for dessert,
roasted, chipped and fried or dried and floured to make a host of confectionaries(Karamura et al.,
1998). Banana flour is utilized in the development of different products such as thin and stiff porridges,
soups, bakery and confectionaries among others. Although bananas and their products are reported to
be rich in starch, sugars and vitamins A and C, potassium, calcium, sodium and
magnesium(Ashokkumar et al., 2018), banana flour is deficient in key nutrients such as proteins,
carotenes and minerals such as iron and zinc. The deficient key nutrients in bananas however, can be

2
obtained from vegetables which are rich in vitamins such as carotenes, folates, and thiamine and
minerals; iron and zinc. The improvement of micronutrient levels of banana flour would consequently
improve its nutrient levels thusa potential to reduce malnutrition levels in children below five years of
age in Uganda. On the other hand, boosting the micronutrient levels of banana flour does not guarantee
the availability of the nutrients. In order to improve the mineral availability, starch solubility and
protein digestibility of the banana-vegetables mix, extrusion technology is the best option. However,
there is limited information on the effect of extrusion conditions (barrel temperature, screw speed and
moisture content) on nutritional properties, physicochemical and acceptability of banana-based flours.
This studytherefore, aimed at formulating and optimizing the extrusion conditions for the production of
instant banana-vegetables soup powder.

1.3. Objectives

1.3.1.Main objective

To optimize the production process for a banana-vegetables instant soup flour.

1.3.2. Specific objectives

1. To optimizethe quality ofsoup formulations fornutritional quality and physicochemical

properties of banana-vegetable soup powder.
2. To optimize extrusion conditions and determine the effect of incorporation of cooked vegetable-
chicken mix on sensory acceptability, nutritional composition and physicochemical properties
of banana-vegetable soup powder.
3. To determine the effect of incorporation of cooked chicken-vegetable mix on shelf-stability of
the instant soup powder.

3
1.4. Hypotheses

1. H 0 ; 1   2  3
Where µ1, µ2, and µ3 show that there are changes in Nutritional composition as a result of changing
proportions of bananas-amaranths and vegetables.

2. H 0 ; 1   2  3
Where µ1, µ2, and µ3 show that there are significant changes in Nutritional quality, physicochemical
properties of foods after extrusion and incorporation of chicken on nutritional quality and acceptability
of the soup.

3. H 0 ; 1   2  3
Where µ1, µ2, and µ3 show that there are significant effects of using chicken on shelf stability of banana
vegetable soup flours.

1.5. Justification of the study

Bananas are seasonal with high post-harvest losses of 22-45%. This results into high nutritional and
economic losses amounting of $411.9 – 842.5 million annually (Muranga et al., 2010). To reduce such
losses, value added products are required. This was increase shelf life of bananas thus ensuring a year-
round availability. Post-harvest losses amounting to 30-40% are also recorded in vegetables (Kiremire
et al., 2010). Pumpkins, tomatoes, mushrooms and carrots were thought important for use in the study
basing on their nutrient composition. Pumpkins are rich sources of minerals such as zinc and iron and
vitamins (Saeleaw & Schleining, 2011). Carrots are good sources of vitamin A and C (Arscott &
Tanumihardjo, 2010). The vegetables have a potential for improving the bananas micronutrients such
as zinc, iron and vitamin A. to improve the nutrient composition of the product and also make the
product a feasible alternative for the fast growing population, extrusion technology was used as a good
alternative to make an instant product. This will serve to reduce the micro nutrient and macro nutrients
malnutrition in the country. In addition, micronutrient and macronutrient malnutrition is recognized by
the Ugandan government as a potential threat to the general health and welfare of the entire population.
Fortification of foods therefore is strongly recommended by the government. The developed products
can potentially contribute towards reducing recorded levels of malnutrition among children below five
years.

4
CHAPTER TWO: LITERATURE REVIEW

2.1. Bananas

The focus of this study was about utilization of Bananas in developing longer shelf stable products.
Understanding bananas is therefore of paramount importance for this study. Bananas belong to Musa
acuminata and Musa balbisiana (Figure 2.1) species. Bananas of the genus Musa are part of the family
Musaceae. They are considered to be derived from the wild species acuminata (AA) and balbisiana
(BB). The plantain, or cooking banana, is classified as Musa paradisiacal. Bananas are mainly tall,
upright, and fairly sturdy. They are often mistaken for treesand has a pseudo-stem which stretches to a
height of up to 2–8 m. The leaves can be of up to 3.5 m in length. Banana fruit grows in hanging
clusters, with up to 20 fruits to a tier (called a hand) and 3-20 tiers to a bunch. The total of the hanging
clusters is known as a bunch and can weigh between 30–50 kg. A single fruit averages 125 g, of which
approximately 75% is water and 25% dry matter content. The edible part of the banana contains, on
average, 75% water, 10% carbohydrates, and about 1% fat, 3% protein and 10% fiber.

Figure 2.1: Classification of Bananas (Karamura et al., 1998)

2.2.1. Banana production in Uganda

Banana is the world‟s fourth most widely consumed crop after maize, rice, and wheat. It is used as a
staple food for most countries in the tropics and subtropics regions. The development of high yield,
short-time growth and disease resistant banana varieties by institutions of agriculture has increased the
5
production volume of banana at harvest. At the global level, banana production is estimated at 72.5
million metric tons per year (Radha Krishna et al., 2012). Six million tonnes are produced annually in
Uganda (Tumutegyereize et al., 2011). This production is from 70% of farmers in Uganda‟s major
producing districts (Bagamba et al., 2007) on approximately 973,340 Ha (Uganda Bureau of Statistics,
2016). These bananas are mainly transported to urban areas, where they are eaten as fruit vegetables.

2.2.2. Physical characteristics of bananas

Bananas are perennial crops with high water succulent stems that give them high resistance to seasonal
droughts. Both Musa acuminata and Musa balbisianavarieties differ in physical characteristics. Both
types mature in about three months from flowering time. The “AAA-EA” Mbidde type of bananas has
compacted bunches of fruit, with starchy banana fingers, but require an expert to differentiate them
from the cooking varieties. The Kisubi banana trees are usually shorter than those of cooking varieties,
whereas the Sweet Ndiizi cultivar has smaller banana fingers and smaller stems in the mother crop.
The average size of a banana fruit is 150 to 200 mm in length, with a diameter of 40 to 60 mm
(Karamura et al., 1998). The cooking variety (plantain) is usually harvested green, while the juice-
yielding type can be harvested at full maturity when the first fingers near the stalk turn yellow. The
ripe fruit has a unique flavor (taste and aroma).

2.2.3. Chemical characteristics of bananas

Chemical properties of cooking banana cultivars are presented in table 2.1. The chemical composition
of bananas varies and the variations are reported to be the result of many factors, including ecological
location, nutrition, location on the bunch from which the banana fingers are sampled for analysis, and
maturity of the fruit at harvest (IITA, 1993). The different cultivars also differ considerably in terms of
their chemical composition with regards to starch, sugar, fat, minerals, acidity, water content, pectin,
and tannins among others. The starch content of ripe and unripe banana fruits lies in the range of 0 -
28.6% (Byarugaba-Bazirake, 2008). Kayisu, Hood, & Vansoest, (1981) reported that unripe bananas
have starch contents of 20 to 25%, although no mention was made of the particular cultivar that was
tested.

The average moisture content in most banana varieties is 75% with approximately 0.5-1% pectin when
ripe. In the banana pulp, insoluble protopectin decreases from about 0.5% to about 0.3% fruit weight
and soluble pectin shows a corresponding increase during ripening(Wills, Lim, & Greenfield, 1984).

6
 Byarugaba-Bazirake, (2008) reported that fully ripe bananas might contain up to one-third to two-thirds
of its pectin in soluble form. Tannins as chemical components of bananas are of great significance for
flavor. When tannins react with proteins and glycoproteins on the surface of the tongue and buccal
mucosa, they cause a drying and puckering sensation known as "astringency".

Table 2.1 Chemical composition of matooke from different cultivars

Chemical Cultivar and chemical composition (%)

composition
Nandigobe (AAA-AE) Bukumu (AAA-AE) Embururu (AAA-AE)
Starch (%) 81.8 82.5 82.9
Protein (%) 4.71 5.1 4.01
Fat (%) 0.87 ND 0.56
Crude fibre (%) 1.25 ND 1.33
Ash (%) 4.34 3.58 4.1
Calcium (µg/L) 0.0058 0.0044 0.0052
Potassium (µg/L) 1.9 1.82 1.84
Magnesium (µg/L) 0.09 0.09 0.01
Tannin (Abs at 500nm) 0.111 0.181 0.012
Source: (Tumutegyereize et al., 2011)

2.2.4. Nutritional value of banana

Bananas are excellent sources of potassium. Potassium can be found in a variety of fruits, vegetables,
and even meats, however, a single banana provides 23% of the potassium that a person needs on a daily
basis. Potassium benefits the muscles as it helps maintain their proper working and prevents muscle
spasms. In addition, recent studies are showing that potassium can help to decrease blood pressure in
individuals who are potassium deficient. Potassium also reduces the risk of stroke. Bananas are
excellent source of vitamins, including vitamin A which aids in healthy teeth, bones, soft tissue, and
more, B6 which aids the body's immune system, promotes brain health, heart health, and more, C which
aids in healing, growth of tissue, ligaments, and more and vitamin D which helps the body to absorb
calcium. In addition, bananas aid in vitality, which makes a person have more energy both mentally and
physically. Bananas are also an excellent food for people who want to lose weight.

7
2.2.5. Banana value chain in Uganda

Annual domestic banana consumption is between 220-460 kg/person and is the highest in the
world(Kiiza et al., 2004). It is the main food staple for some 13 million people and an important source
of food, nutrition, and income security for smallholder producers. Producers consume about 70% of
harvested bananas at home. Banana consumption (and production) is concentrated in the central and
western regions with the latter having the highest consumption. Consumption is least in the northern
region (figure 2.2). Processing of bananas into value added products takes the least share and stands at
3%. This can depict the amount of post-harvest losses in faced by bananas since shelf stable products
out of bananas are low. Thus, processing of bananas into value added products is one area that is
lacking.

Figure 2.2: Banana end markets in Uganda and consumption (Source: Bill and Melinda gates
foundation report, 2009)

2.2.6. Processing of bananas into value added products

Bananas can be processed into chips, raw ripened fruit, cooked green banana, fermented and
unfermented beverages, juice, puree, and dried flour for bakery and infant formula food (Mohapatra,
Mishra, & Sutar, 2010) as shown in figure 2.3. Banana is also used as a starch source for various
chemicals and packaging materials.

8
 Figure 2.3: Flow chart showing processing of bananas into different products(Mohapatra et al.,
2010)

In Uganda, the Presidential Initiative for Banana Industrial Development (PIBID) located in western
region is the leading head in banana processing. PIBID gets banana flour from matooke to increase
shelf-life of fresh bananas. According toMuranga et al., (2010), Matooke (Green banana) is a very
starchy food, compared to maize and potatoes, thus produces a better product. Banana flour can also be
used as an alternative to wheat for its good starch composition in confectionaries.

In Kampala, a supermarket survey concluded that some available products include “Ndizi” crisps from
apple bananas. These are made from locally grown bananas and plantain or “gonja”. Another
available productcalled Fantazee banana brunch, is manufactured by Deepa Industries in Kenya.
Since most bananas are produced for traditional cooking and brewing at home, bananas are seen as
subsistence fruits. Despite being the world's second largest producer of bananas (after India), Uganda
does not feature among the important countries trading bananas or their products on the world market.
This has been greatly contributed by low shelf life and high post-harvest losses.The feasible alternative
to make products from bananas would propose improving shelf-life as well as reducing post-harvest
9
losses thus calling for value addition. Development ofsoups from cooking bananas is a new area of
interest for research.

2.2. Description of soups

Soup is defined as a liquid food made by stewing ingredients such as vegetables, meat, fish often in a
stock and with seasoning (Fatima & Al-Subhi, 2013). Hot soups are additionally characterized by
boiling solid ingredients in liquids in a pot until the flavors are extracted, forming a broth.Traditionally,
soups are classified into two main groups, clear soups and thick soups (Radha, et al., 2015). Clear
soups are based on clear un thickened broth or stock. Clear soups can be served plane or with a variety
of vegetables or meat. Thick soups are classified depending upon the type of thickening agents
used.Purées are made from vegetables thickened with starch.Bisques are made from puréed shellfish or
vegetables thickened with cream. Cream soups can be thickened with eggs, butter, and cream. Other
ingredients commonly used to thicken soups and broths include eggs, rice, lentils, flour, and grains.
Other popular soups also include carrots and potatoes soup. In the interest of this study, thick soups
were produced. Thick soups were made from banana flour, Amaranths supplemented with either
tomatoes, mushrooms, pumpkins or carrots or all of them. The thickening agent was starch from
bananas.

2.2.1. Commercial soups

Commercial soup became popular after the invention of canning in the 19th century. These soups can
take two forms: canned soups and dried soups.

2.3.1.1.Canned

Canned soup can be condensed, in which case it is prepared by adding water (or sometimes milk).
Canned soup can be prepared by heating in a pan, on the stovetop or in the microwave. Such soups can
be used as a base for homemade soups, with the consumer adding anything from a few vegetables to
eggs, meat, cream or pasta. Condensing soup allows soup to be packaged into a smaller can and sold at
a lower price than other canned soups. Since the 1990s, the canned soup market has burgeoned with
soups marketed as ready-to-eat, which require no additional liquid to prepare. Microwaveable bowls
have expanded the ready-to-eat canned soup market, even more, offering convenience and are popular
lunch items.

10
2.3.1.2.Dried

Dry soup mixes are sold by many manufacturers and are easy to reconstitute with hot water. During
preparation, other fresh ingredients may be added. The first dried soup was bouillon cubes. East Asian-
style instant noodle soups include ramen and seasonings. They are marketed as a convenient and
inexpensive instant meal, requiring only hot water for preparation. Other dried soups include vegetable,
chicken base, potato, pasta and cheese flavors.

The soups analysis shown above requires that dried soups are durable and more convenient than canned
soups. They have longer shelf-life than canned soups. In this study therefore, dried soups are preferred.
Dried soups were developed from dried banana flour together with other vegetables. Grain amaranth
were used to improve the protein value of bananas. Vegetables chosen include carrots, pumpkins,
tomatoes and mushrooms. Unnecessary vegetables wereeliminated by carrying out a response surface
modelling using design expert version 11.

2.3. Choice of supplements to banana flour…for.

Provision of a nutrient-rich mixture from banana flour requires supplements. Vegetables have been
chosen for their availability and a good nutrient base.Mushrooms, pumpkins, tomatoes and carrots, and
amaranth grains wereused. Knowledge about these vegetables and grains is paramount.

2.3.1. Grain Amaranth

Grain amaranth belongs to order Caryophyllales, family Amaranthaceae, sub-family Amaranthoideae,

genus Amaranthus, and tothe section Amaranthus. The genus Amaranthusincludes approximately 60
species. Most of these species are cosmopolitan weedswhile others are cultivated amaranth species. The
cultivated species can be used as food grain, leafy vegetables, forage and ornamentals(Mlakar, et al.,
2010). The principal species considered for grain production include Amaranthushypochondriacus, A.
cruentus and A. caudatus.Amaranth is a highly nutritious grain; the seeds of A. hypochondriacus
contain 15-20%of lysine-rich protein (3.2-6.4 g/100g protein compared to 2.8-3.0 g/100g protein for
wheat), 58-66% of starch, 6-9% of raw fiber and 6-8% of highly unsaturated lipids. Amaranth has high
concentrations of calcium (250 mg/100g) and iron (15 mg/100 g), which areten and four times higher,
respectively, than those found in wheat (Milán-Carrillo et al., 2012). Amaranth is an excellent source of
iron (Table 2.2).

11
2.3.2. Pumpkin

Pumpkin belongs to genus Cucurbitaof the family Cucurbitaceae,whichis one of the largest families of
the plant kingdom(Bhat & Bhat, 2013). Pumpkinsare widely grown and consumed in many tropical and
sub-tropical countries around the world. Pumpkins are found in many shapes, sizes and colors.
Pumpkins are consumed in a variety of ways such as fresh or cooked vegetables. Pumpkin can be
processed into flour which has a longer shelf-life. Pumpkin flour is used because of its highly-desirable
flavor, sweetness and deep yellow-orange color. It has been reported to be used to supplement cereal
flours for soups, sauces, instant noodle and spice as well as a natural coloring agent in pasta and flour
mixes(Kuchtová et al., 2016). Pumpkin is rich in β-carotene (Table 2.2), vitamins, minerals, pectin and
dietary fiber(Saeleaw & Schleining, 2011). Consumption of foods containing carotene helps prevent
vitamin deficiencies such as skin diseases, eye disorders etc. Incorporation of β-carotene rich materials
in human diet is therefore considered a cost-effective approach to vitamin-A related health problems
(Pongjanta et al., 2006).
Pumpkin flours have been utilized to produce vitamin A rich soups(Ravi, Lakshmi, & Ranjani, 2010a)
reported an increase in the β-carotene content of complementary weaning mix based on sorghum,
green-gram and rice flour on incorporation of pumpkin flour. Swar, Alhaj, & Osman, (2014) reported
that a vitamin A rich porridge can be obtained from blends of sorghum flour and pumpkin pulp to
address vitamin A deficiency in Sudan. In Ethiopia, Abebe et al., (2011) found out that addition of
pumpkin to corn-legume blend and Kocho legume blend increased the vitamin A content of the mixes
by 25 and 180-fold respectively.

2.3.3. Carrot

Carrot (Daucuscarota L) is one of the popular root vegetables grown throughout the world. China is the
major carrot producing country in the world (Sharma, Karki, Thakur, & Attri, 2012). Carrot is an
economically important horticultural crop that has gained popularity in recent decades due to increased
awareness of its nutritional value (Arscott & Tanumihardjo, 2010). Carrots contribute significantly to
dietary vitamin A intake through α- and β-carotene (Table 2.2) and modestly to other nutrients (Arscott
& Tanumihardjo, 2010). Carotenoids are important micronutrients for human health. The total
carotenoids content in the edible portion of carrot range from 6000 to 54,800µg/100g(Sharma et al.,
2012).In India, carrot powder has been incorporated into dal soup to address vitamin A deficiency

12
(Pandey & Kulshrestha, 2003).Onabanjo et al., (2008) blended carrot powder in addition to other
ingredients to develop a complementary food.

Table 2.2: The nutritional composition of grain amaranth and some selected vegetables (Value
per 100g)
Nutrient Grain Mushrooms Tomatoes Carrot Pumpkin
amaranth

Energy (Kcal) 371.00 328 302 341 364

Protein (g) 13.56 10.0 12.91 8.10 14.29
Lipids (g) 7.02 4.0 0.44 1.49 3.57
Carbohydrate* (g) 65.25 64.66 74.68 79.57 71.43
Iron (mg) 7.61 1.00 4.56 3.93 4.50
Zinc (mg) 2.87 1.54 1.71 1.57 0.32
Vitamin A (µg RAE) 0.00 0.00 862 3423 0.00
Sugars (g) 1.69 3.26 43.9 38.82 28.57
Fiber, dietary (g) 6.70 4.8 16.5 23.6 14.3
Vitamin C (mg) 4.20 7.0 116.7 14.6 0.00

*Content determined by difference Source: USDA (2018)

2.3.4. Mushrooms

Mushrooms are healthy foods. They are poor in energy content and fat but are rich in vegetable
proteins, chitin, vitamins and minerals (Manzi et al., 1999). Mushrooms have a great nutritional value
since they are quite rich in protein. They are sources of important content of essential amino acids and
fiber, with poor fat content(Reis et al., 2012). Edible mushrooms are rich sources of vitamins (B1, B2,
B12, C, D and E) (Reis et al., 2012).Mushrooms have also been reported as therapeutic foods, useful in
preventing diseases such as hypertension, hypercholesterolemia and cancer (Manzi, Aguzzi, and
Pizzoferrato, 2001). The nutritional content of mushrooms is indicated in table 2.2 above.
2.3.5. Tomatoes

Known scientifically as Solanumlycopersicum, tomato is the berry of a plant from the nightshade
family. It is native to South America. Despite technically being a fruit, tomato is generally categorized
as a vegetable.Tomatoes are the major dietary source of antioxidant lycopene, which has been linked to

13
many health benefits, including reduced risk of heart disease and cancer.They are also a great source of
vitamin C, potassium, folate and vitamin K.They are usually red when mature, but can come in a
variety of colors, including yellow, orange, green and purple.Then there are many subspecies of
tomatoes, with different shapes and flavor.
The combinations of different compositions of soups are thought to supply the nutritional demands of
children aged 1-5 years shown in table 2.3.

Table 2.3: Recommended daily Nutritional requirements for 2-5 years old children

Category Age (yrs) Protein (g) Fat (g) Energy Calcium Iron (mg)
(kCal) (mg)

Children 2-3 16.7 27 1060 600 9

Children 4-6 20.1 25 1350 600 13

The correct quantity of different proportions that can give the daily nutritional requirements of children
aged 2-5 years wasobtained by carrying out response surface modeling. Response surface modeling can
be effective when the process is carefully thought about through optimization of different ingredients.
There is need to understand the process of optimization.

2.4. Process optimization

2.3.6. Background

Optimization improves performance of a system, process, or product with an objective of obtaining

maximum benefit(De Aguiar et al., 1995). Issues with optimization usually arise in food production
and processing while designing, developing and managing quality of food products. Optimization of
food productionseeks to provide answers to the following questions.

1. How changes in one set of variables affect overall food quality? How are they correlated with
each other?

2. Which food raw materials and recipes should be used to produce an optimal quality food
product?

3. Which processing conditions must be used to getthe best food product?

4. How to determine the cause of food product defects and further control them?

14
 5. Are the recipes and processing conditions currently used for an existing food product the best?

6. What measures can be taken to increase production yield?

7. Is it possible to have a successful food product with cost-effective raw materials and
procedures?

2.3.7. Optimization methods

There is a myriad optimization approaches which can be employed in process design. These include
fractal analysis, Response Surface Methodology (RSM), generic algorithms, multivariate analysis,
fuzzy logic and linear programming (Sablani, 2008). Majority of the conventional approaches are
arduous and time-consuming since they involve numerous runs. Interaction factors are also ignored and
thus chances of obtaining optimum conditions are uncertain (Mu, et al., 2009). Additionally, some of
these conventional methods like generic algorithms and fuzzy logic are very complex and not user-
friendly. Additionally, there is limited knowledge on the use of these optimization approaches hence
their limited application (Sablani, 2008)

2.3.8. Choice of an optimization method

Response Surface Methodology (RSM) is the commonest and most effective tool used in the food
industry and biotechnology for optimizing processes (Bas and Boyaci, 2007). RSM involves a
collection of statistical and mathematical techniques that are useful for developing, improving, and
optimizing processes in which a response of interest is influenced by several variables (Montgomery,
2001). The objective is to simultaneously optimise levels of the variables to generate best system
performance (Box & Draper, 2007)).

RSM enables rapid, effective and efficient development of new products and process at a reduced cost
and time (Hu, 1999; Peričinet al, 2008). RSM reduces the number of experiments required even when a
myriad of variables is involved and this makes it less laborious and time-consuming than other methods
(Wang et al, 2014). RSM also facilitates identification of operating conditions that favour high yields
and production of certain products. RSM is flexible and widely used in production and optimization of
different products (Singh et al., 2009).

15
2.3.9. Experimental designs in RSM

It is very important to select an experimental design before applying RSM. There are many types of
experimental designs. The choice depends on the complexity of the model and whether models
sufficiently represent the experimental data adequately. Examples of these experimental designs
include three level factorials, Box–Behnken, central composite, optimal and Doehlert designs (Myers et
al., 2009). The designs differ from one another with respect to the number of levels of variables, runs,
and blocks. However, Central Composite Design (CCD) and d-optimal are the most popular among the
RSM designs (Bezerraet al., 2008). The d-optimal design is commonly used in designing experiments
with two quantitative factors having three levels each. D-optimal design minimizes the generalized
variance of the estimated regression coefficients (Unalet al, 1998). Thus, offers an added advantage
compared to CCD.

2.3.10. Steps involved in the application of response surface methodology

According to Montgomery, (2001)the steps followed during optimization using RSM include:

1. Selection of independent variables of major effects on the system through reviewing literature and
the restriction of the experimental region, according to the objective of the study.

2. Choosing the experimental design and carrying out the experiments according to the selected
experimental matrix.

3. Mathematical–statistical treatment of the obtained experimental data through the fit of a polynomial
function.

4. Evaluating the model's fitness and model adequacy.

5. Verifying the requirement and possibility of performing a displacement in the direction of the
optimal region.

6. Obtaining the optimum values for each studied variable.

2.3.11. Determination of optimum conditions for multiple variable responses in soups.

Several optimization methods can be used to optimize multi-response systems. The selection of an
optimization method depends on the complexity, merits of the method and recommendations by other
researchers. The different optimization methods include conventional graphics method, nonlinear
programming methods, improved graphics method, extended response surface and desirability function
16
approach (Sablani, 2008).Desirability Function Approach (DFA) is the most important and widely used
in optimization. DFA is an analytical technique for optimizing multiple responses simultaneously. DFA
calculates individual desirability associated with each response and then an overall desirability (ranging
between 0 and 1) can be calculated as the geometric mean of individual desirability. According to
Banga et al. (2003), desirability level indicates the closeness of the response to the set target value.
DFA is simple and quick to apply and allows subjective judgment on the importance of response
variables compared to other techniques including comparing responses of different scales
(Guillou&Floros, 1993). The different formulations were developed from which optimization of the
responses could be determined. The processes used to make instant products is extrusion. There is need
to understand the process of extrusion.

2.5. Extrusion

Extrusion cooking, as a multi-step, multi-functional and thermal/mechanical process has permitted a

large number of food applications (Singh et al., 2007). It has created a huge impact in the food
industries towards shaping and deriving ready to eat products (Fellows, 2000). The use of extrusion in
the food processing has increased its popularity due to its versatility, cost-effectiveness, environmental
friendliness and better product output (Sundarrajan, 2014). The principle of extrusion involves loading
of raw materials in the feeding hopper from where the screw conveys them through a barrel under
pressure. The materials are compressed into a semi-solid, plasticized mass as they pass down the barrel.
The selection of right extruder for the production of ready to eat (RTE) or cereal snacks depends on the
nature of raw materials used, bulk density and type of product to be produced (Fellows, 2000).
2.3.12. Types of extruders

Two types of extruders are used for food production: single-screw extruders and twin-screw extruders.
Single-screw extruders are the most common extruders used in the food industry. Twin-screw extruders
are used for high-moisture extrusion, products that include higher quantities of components such as
fibers, fats, etc. and more sophisticated products (Steel et al., 2012).
2.3.12.1. Single-screw extruders

Single-screw extruders are the most common extruders applied in the food industry. The classification
of single-screw extruders can be defined based on processes involved or equipment parameters such as
conditioning moisture content (dry or wet), solid or segmented screw, desired degree of shear and heat
source. However, practically, the main classification used considers the degree of shear and the heat

17
source (Steel et al., 2012). Single-screw extruders can be grouped into four different types based on the
degree of shear, as follows: cold forming extruders, high-pressure forming extruders, low-shear
cooking extruders and collet extruders (Steel et al., 2012)
2.3.12.2. Twin screw extruders

Twin-screw extruders are composed of two axes that rotate inside a single barrel; usually, the internal
surface of the barrel of twin-screw extruders is smooth. Depending on the position of the screws and
their direction of rotation, four different types of configurations are possible: (i) co-rotating
intermeshing screws; (ii) co-rotating non-intermeshing screws; (iii) counter-rotating intermeshing
screws; and (iv) Counter-rotating non-intermeshing screws. Conical intermeshing extruders also exist
(Steel et al., 2012). The advantage of using twin screw extruders is versatility to process a wide range
of products like tortillas, cereal snacks, extruded corn snacks, and multigrain snacks. Due to high
capital and maintenance costs, single screw extruders are considered to be cost-effective when
compared to twin screw extruders (Sundarrajan, 2014). Figure 2.4 shows a schematic diagram of a
single screw extruder

Figure 2.4: A cross-section of a single-screw extrusion-cooker:

1 - engine, 2 - feeder, 3 - cooling jacket, 4 - thermocouple, 5 – screw, 6 - barrel, 7 - heating jacket, 8 -

head, 9 - net, 10 -cutter, I - transport section, II – compression section, III –melting and plasticization
section (Moscicki et al., 2013).
2.3.13. Processing parameters affecting the nutritional quality of extruded products

Different processing parameters have effects on the nutritional quality of extruded products. The
primary process parameters include feed rate, screw speed, barrel temperature, moisture content, feed
formulation, screw and die configuration. The secondary process parameters include dye temperature,

18
pressure, and torque (Sundarrajan, 2014). Razzaq et al., (2012) studied the effect of barrel temperature,
screw speed and moisture content on the chemical characteristics of maize (Zea mays. L) extrudates.
The author found out that these extruder variables (150 °C, 170 rpm and 18% moisture content)
considerably enhances apparent absorption of sodium, potassium, calcium, and iron. Increase in barrel
temperature and feed moisture content causes an increase in the water absorption index (Agathian et
al., 2015). Several other workers have studied the effect of extrusion variables such as feed moisture,
barrel temperature, screw speed, feed rate (Albarracín, et al., 2015; Dias-Paes and Maga, 2004; Filli et
al., 2010; Ilo et al., 1999).
2.3.13.1. Moisture content

Moisture acts as a plasticizer in extrusion cooking. Several workers have investigated the effect of feed
moisture content in extrusion cooking especially its effect on functional properties of the extrudate.
Bhise et al., (2013) while studying the optimization of extrusion conditions for production of texturized
flaxseed defatted meal by response surface methodology discovered that increase in feed moisture
content causes an increase in fat absorption capacity, water holding capacity, water solubility index and
water absorption index of the flour. According to Chávez-Jáuregui et al., (2000), the study of on the
extrusion cooking process for amaranth (AmaranthuscaudatusL.) showed that increase in feed moisture
caused an increase in shearing stress, density, and a decrease in texture acceptability of the amaranth
flour extrudate. Investigations carried out by Gbenyi, et al., (2017) to evaluate the effect of feed
moisture, barrel temperature and single screw extrusion on physicochemical properties and
acceptability of grain sorghum showed that increase in feed moisture content and die temperature cause
an increase in water absorption index, water solubility index and swelling power.
2.3.13.2. Barrel Temperature

The process temperature is an important parameter in extrusion cooking process. Ilo et al., (1999) while
investigating the extrusion cooking and product properties of rice flour and banana-amaranth blends
discovered that increasing extrusion barrel temperature had a positive effect in the texture softening,
but it increased the browning of the extrudate and probably the extent of nutritional loss due to Maillard
reactions. Singh, et al., 2007 analyzed the effects of extrusion temperature (164, 182, 200°C) levels (0-
30%) and grits moisture content (g/100 g sample) (18-24%) on textural and physicochemical properties
of rice grits.

19
2.3.14. Effect of extrusion cooking on nutritional quality

2.3.14.1. Protein retention and digestibility

Extrusion improves protein digestibility through denaturation, which exposes enzyme-access sites
(Zarzycki et al., 2015). Most proteins such as enzymes and enzyme inhibitors lose activity due to
denaturation. The extent of denaturation is typically assessed as change in protein solubility in water or
aqueous solutions. These changes are more pronounced under high shear extrusion conditions
(Zarzycki et al., 2015). Koeppe et al., (1987) found a slight increase (from 82.1% to 85.4%) in the in-
vitro protein digestibility of banana-amaranth-maize gluten meal extrudates. Extrusion cooking
increased the in-vitro protein digestibility (IVPD) of African bread fruit-defatted soybean-corn flours
by 22.77% (James & Nwabueze, 2013). Rampersad et al., (2003) investigated the effect of extrusion
cooking (barrel temperature of 130 °C, screw speed of 150 rpm and feed moisture 15%) and roasting
on protein digestibility using the in-vitro method. The authors discovered extrusion cooking improved
the in-vitro protein digestibility (IVPD) of the pearl millet based weaning foods.

2.3.14.2. Vitamin A retention

Vitamin A deficiency is a major cause of blindness in many less-developed nations, and the vitamin is
important for healthy immune system function. Unfortunately, oxygen and heat destroy vitamin A and
related carotenoids. Increasing barrel temperatures from 125 to 200ºC resulted in more than 50%
destruction of trans β-carotene in wheat flour 50% (Zarzycki et al., 2015). Extrusion cooking (25 %
moisture content, 120 rpm and 100 °C) caused a significant decrease of vitamin A contents of extruded
blend of pigeon pea and unripe plantain blends (Rampersad et al., 2003). James and Nwabueze, (2013)
reported 65.95% vitamin A retention of an extruded full-fat blend of African breadfruit soybean-corn
snack at 21% moisture content and 140 °C barrel temperature.
2.3.14.3. Mineral bioavailability

Extrusion can improve the absorption of minerals by reducing other factors that inhibit absorption.
Extrusion increases the apparent absorption of iron in extruded meal-based diets (Alonso, Rubio,
Muzquiz, & Marzo, 2001). Poltronieri et al., (2000) assessed the effect of extrusion cooking on the in-
vitro iron availability of African weaning foods; their results show that extrusion increases the available
iron.

20
2.6. Instant products

Instant products are a group of dried foods which play an important role in the nutrition of people
because they can fulfill present and future social consumer requirements. Instant products come in
diverse forms which aid easy access to nutrients (Usha, 2007). Pelembe et al., (2002) developed a
protein-rich composite sorghum–cowpea instant soups by extrusion cooking process. Instant grain base
for use in weaning foods was made from a blend of wheat, mung bean and groundnut (Pathania et al.,
2013). Milán-Carrillo et al., (2012) produced high antioxidant instant banana-amaranth flour suitable to
elaborate a nutraceutical beverage. Shadan, et al., (2014) formulated, prepared and evaluated low-cost
instant products based on cereals and pulses.

2.7. Sensoryevaluation

Sensory evaluation refers to the science of judging and evaluating quality of food by the use of senses.
It is of practical application in product development and helps in product matching, improvements,
grading as well as establishing the worth of acceptance of a product(Meilgaard, Sc, Civille, & Carr,
1999). Sensory attributes of a food product are important in its overall acceptance (Cardello, 1994).
The acceptability of any food is largely influenced by its organoleptic properties. These sensory
attributes are a function of the physico-chemical composition of food product. They are influenced by
quality and composition of raw materials (Arvanitoyannis et al., 2007).In the process of sensory
evaluation, food is uniformly prepared and presented to panelists in isolated booths (Hashmi, 2007).
Panelists evaluate the food and recordscores for parameters as required in the study on a sensory
evaluation sheet that is decoded and analyzed by statistical procedures. The parameters commonly
evaluated include taste, mouth feel, flavor, texture, color, aftertaste, appearance and overall
acceptability.

A consumer acceptability taste using a 9-point hedonic scale is used to assess and compares the sensory
attributes of the samples provided. The commonly used hedonic scale is; “1” Dislike extremely, “2”
Dislike very much, “3” Dislike moderately, “4” Dislike slightly, “5” Neither like nor dislike, “6” Like
slightly, “7” Like moderately, “8” Like very much and “9” Like extremely

2.8. Shelf life studies of food products

Food is inherently perishable and depending on its physical and chemical properties and storage
conditions, there is a point when either its quality becomes unacceptable or harmful to the consumer.

21
At this point, it is said to have reached the end of its shelf-life and the ability to predict this is of great
value to the food industry. According to NZFSA (2005), the shelf life of a product begins from the time
the food is manufactured. Most consumers on daily basis not only make demands as to how to obtain
high-quality foods but also have some expectations that such qualities obtained from these foods was
maintained at a high level during the period between purchase and consumption (Kilcast and
Subramaniam, 2000)

2.3.15. Definition of shelf life

IFST (1993) defines product shelf-life as the time during which the product was remain safe, be certain
to retain desired sensory, chemical, physical and microbiological characteristics and comply with any
label declaration of nutritional data when stored under the recommended conditions. Thus, shelf life of
a dried soup would be the time during which the product remains safe, retains its color, flavor, and
aroma, have a stable peroxide value, free fatty acid content as well as be microbiologically safe and
stable.

2.3.16. Factors affecting shelf life

Product shelf life is controlled by three major factors; product characteristics, environment to which the
packaged product is exposed during distribution and properties of the package (Robertson, 1993)

2.3.16.1. Product characteristics

According to Robertson (1993), most foods fall into one of these three main classes; perishable (very
short shelf life products), semi-perishable (short to medium shelf life products) and ambient
temperature shelf stable (medium to long shelf life products). These are based on different
characteristics associated with different food products as well as kinds of changes that occur during
their storage. Ambient temperature shelf-stable foods include some natural foods such as cereal grains,
nuts and confectionery products and processed food products such as canned foods, soft drinks and
cake mixes.Thus, soups fall into this category.

2.3.16.2. Distribution environment

Packaged foods may lose or gain moisture; they could also reflect the temperature of their environment
because very few food packages are good thermal insulators. Thus, the environment has an important
influence on the rate of deterioration of packaged foods.According to IFST (1993), factors that

22
influence or control the shelf-life of food products can be again grouped into intrinsic and extrinsic
factors.

Intrinsic factors

Intrinsic factors are properties of the final product which include water activity (aw); pH value and
total acidity (type of acid); available oxygen; natural micro-flora and surviving microbiological counts;
natural biochemistry of the product formulation (enzymes, chemical reactants) and the use of
preservatives in product formulation (for example salt or sodium bicarbonate).

Extrinsic factors

These are factors which the final product encounters as it travels through the food chain. They include
time-temperature profile during processing; temperature control during storage and distribution;
environmental microbial counts during processing, storage, distribution and consumer handling.

2.3.17. Predicting shelf-life/shelf life testing

Introducing a new food product on the market usually does not take a day. Certain measures especially
concerning shelf-life need to be established before certification to be put on shelves for consumers.
Thus, food products require a date or labeling indicating when the product is likely to become
unacceptable for consumption. As stated by Kilcast and Subramaniam (2000), while determining shelf
life of products, products which may be expected to have a longer shelf life may require knowledge of
their storage characteristics over the intended shelf-life period and this can introduce unacceptable
delays in putting the product on the shelf.

Generally, the shelf life testing of food products falls into one of three categories (Gacula & Kubala,
1975)

 experiments designed to determine the shelf life of existing products

 experiments designed to study the effect of specific factors and combinations of factors such as
storage temperature, package materials, or food additives on product shelf life

 tests designed to determine the shelf life of prototype or newly developed products

23
2.3.18. Selection criteria to assess shelf life

Every food product is associated with specific attributes either in their raw state or after certain
conditions have been applied. Depending on the attributes, certain changes are noticed especially
during storage and these changes help to detect if products have gone bad or not. According to Fuller et
al., (2006), in the selection of criteria for shelf life assessment, two main steps are needed. First,
changes that can be measured and appropriate for the product are selected. These criteria normally
change gradually with time so the onset of the change can be measured. Example of such criteria
includes microbiological, nutritional change, undesirable change (loss or change of color, textural
changes like loss of crispness, staling) and change in a functional property. Second, there must be some
decision made about how much loss of quality characteristics can be accepted. In other words, how
much loss of a quality characteristic can be accepted before spoilage is declared

2.3.19. Types of shelf life testing

According to NZFSA, (2005), there are two main methods of shelf life analysis. The two methods need
to be exploited for comparison.

2.3.19.1. Direct method:

This is the most commonly used testing method. It involves storing the product under the normal or
pre-selected conditions for a period of time longer than the expected shelf life. The product is checked
at regular intervals to see when it begins to spoil. The exact procedure is unique for each product. This
method is usually referred to as real-time studies. Shelf life of food samples can be determined
according to the flow chart illustrated in figure 2.5 below.

Before product on market Identify what may Decide which tests to use
causefood spoilage

Once product on market Determine shelf life Plan the shelf life study

Continue to monitor shelf life

24
 Figure 2.5: Flow chart illustrating steps for calculating the shelf life of food products by direct
method (Source: NZFSA, 2005)

2.3.19.2. Accelerated shelf life studies

This attempts to predict the shelf life of a product without running a full-length storage trial. This
typeof study is usually used for a product with a longer shelf life. Acceleration factors such as
temperatureare applied to the product to attempt to increase the rate of deterioration.The
experimentaldetermination of shelf-life using direct method can require a considerable amount
ofexperimentation, with consequent costs and demands on time. When the actual storage time islong,
shelf-life studies are based on accelerated shelf life testing (ASLT). This techniqueshortens the process
of obtaining theexperimental data (Kebedeet al., 2015; Mizrahi, 2000). With accelerated shelf life
testing the trialperiod is shortened by deliberately increasing the rateof deterioration (Martins and Silva,
2004). This is usually done by increasing the storagetemperature. The results are then used to estimate
the shelf life under normal storage conditions(Yang et al., 2013).With accelerated shelf life testing the
trial period is shortened by deliberately increasing the rate of deterioration (Martins and Silva, 2004).
This isusually done by increasing the storage temperature. The results are then used to estimate the
shelf life under normal storage conditions(Yang et al., 2013).
Accelerated shelf-life testing (ASLT) methods have been used to predict shelf-life at normalconditions
(Rao et al., 2012) based on data collected at high temperature or high humidityconditions (Achouretal.,
2001). The data from higher testing temperatures in food quality lossare used to determine shelf-life at
regular storage conditions through the use of Arrhenius orLinear equations (Gulla and Waghray, 2012).
Food products are conditioned and stored atelevated temperature and/or humidity and the quality
changes of the product are evaluated at aspecific sampling rate (Achouret al., 2001). ASL study could
significantly shorten the durationof shelf life study to half or quarter of the direct method (Cardelli and
Labuza, 2001). One of theprincipal methods of predicting the shelf life of processed food products is to
monitor the level oflipid degradation in fat containing foods stored at elevated temperatures. The
quantitative approach to shelf life prediction requires that the deteriorativemechanisms limiting shelf
life of the specific food be identified and that the index of deteriorativereaction be measured as a
function of time. Since the chemical reactions in food can be very complex it is usually easier to

25
examine a reaction from purely mathematical or semi empirical approach based onchemical laws rather
than on a mechanistic approach in which each step must be known(Labuza and Kamman, 1983).
The loss of quality for most foods can be presented by a mathematical equation of the followingform:
dA
  KA n where A= Quality factor measured
d
 =time
K = a constant which depends on temperature and water activity
n = a power factor called order of the reaction which defines whether the rate isdependent
orindependent.
dA
 = rate of change of time. A negative sign is used if the deterioration is loss of A andpositive if itis
d
for production of undesirable products.
For quality a change in foods, the reaction order has generally been shown to be either 0 or 1depending
on the reaction involved (Labuza, 1982; Pope, 1980). If n = 0 the reaction is said to bezero order with
respect to A. This means that the rate of loss of A is constant and independent ofthe concentration of A.
Zero order reaction is mainly applicable to non-enzymatic browning indried foods and lipid oxidation
in dried foods (Labuza, 1982).
When n = 1 the reaction is first order whereas the loss of quality is dependent on the amount of A left.
The deterioration which falls in first order reaction includes vitamin and protein loss in dried foods
and vegetable rancidity in dried foods (Labuza, 1982).In some studies reactions were describedby
nonlinear equations such as polynomial equations (Smoot and Nagy, 1980).
The principle factor affecting the kinetics of reactions in processed and dried foods istemperature.
Increases in temperature are known to accelerate deteriorative reactions in food andthus reduce shelf
life of food. According to Labuza (1982), temperature is assumed to follow Arrhenius equation
aspresented in equation 2.1.

K  K o exp (Ea/RT) 2.1

Where
K= rate constant
Ko= pre exponential constant
Ea= Activation Energy
R = Gas constant
26
T = Absolute temperature
Arrhenius equation is the best approach in modeling temperature dependence (Saguy and Karel,
1980), since unlike other temperature dependent models, it based on theories of thermodynamics
(Labuza and Kamman, 1983). The activation energy is generally derived from the slope of a plot of
natural logarithm of rate of constant (k) versus the inverse of absolute temperature and depends on
compositional factors such as water activity, moisture content and solid concentration. When the
reaction mechanism changes with temperature the activation energy may vary substantially.
Large statistical errors are commonly associated with the calculations of the temperaturedependence of
the reactions. Some studies have suggested methods of analyzing kinetic datawhich provide statistically
more reliable results (Cohen and Saguy, 1985). Arrhenius plot canalso be used to establishshelf life
plots of specific products based on a known end point qualitydeterioration value (Labuza and
Kamman, 1983). There could be many limitations besidesstatistical errors in using the Arrhenius and
shelf life plots to predict shelf life at lowertemperature. Generally the problem occurs because some
reactions which predominate at highertemperature do not pre dominate at lower temperature (Labuza
and Riboh, 1982).
Another important prediction for accelerating the effect of EA called the Q10 Value is also used
(Leeand Krochta, 2002). The relationship between Q10 and EA is described in equation 2.2 below

2.189 E A  shelf life at T  10 

log Q10   log   2.2
T (T  10)  Shel life at T 
Where Q10 Increase in rate or decrease in shelf life for a 10oC rise in T
T is in K
EA is in Cal/mole
2.3.20. Measurement of product shelf-life

2.3.20.1. Sensory panels

Sensory techniques are required in determinations of changes in food quality as they are subjected to
storage (Kilcast and Subramaniam, 2000). Trained panelists are commonly used to measure the sensory
changes in a product over time. The panelists should be highly skilled at describing the appearance,
aroma, flavor, and texture of the product. For most shelf life studies, three to five trained sensory
panelists evaluate the samples at each time points. A sensory panel leader would lead the group
discussion and record their consensus of opinion. However, the final decision on the acceptability of
the product is reserved to the project head or the company (National foods lab, 2013).
27
2.3.20.2. Physical measurements

Changes in color and texture are the common physical measurements used to monitor changes in food
product attributes during storage. Thus, a change in product color or texture can be used to detect
whether the product has gone bad or is still in good condition for human consumption. These changes
may be as a result of some chemical reactions occurring in the product, such as those caused by
interaction of ingredients or by environmental influences, such as moisture migration through the
packaging and non-enzymatic reaction which may alter the color of the product Kilcast and
Subramaniam, (2000)

2.3.20.3. Chemical measurements

Chemical analyses play a vital role in shelf-life testing. This is because they can be used either to
measure the endpoints of chemical reactions occurring in food during storage or to confirm the results
obtained by the sensory panel Kilcast and Subramaniam, (2000). Even though there are many chemical
reactions occurring concurrently in any food product especially during storage, only the major ones
influencing changes in the product quality are monitored and measured during shelf-life testing Kilcast
and Subramaniam, (2000). Some chemical tests used in determining changes in a particular quality
characteristic include free fatty acid and peroxide value measurements which serve as markers for the
level of rancidity of products.

2.3.20.4. Microbiological stability determinations.

According to Kilcast and Subramaniam, (2000), important aspects considered in the determining of
microbiological stability of food product are;

1. Microbial growth, which leads to the spoilage of a food product

2. The growth of microbial pathogens that affect the safety of the product

The water activity, storage temperature, time and pH are used to predict to a large extent the micro-
organisms that are likely to grow in a given food product. The time to spoilage is usually determined by
storing the product at the appropriate temperature and measuring the microbial load at regular intervals.
The time to reach a pre-determined level of microbial count (total count and level of individual
microbes) is considered to be the end-point. Since it is advisable to leave a safety margin in setting the

28
shelf-life, generally 70% of the time to spoilage is taken to be the storage life (Gonzallez-Ferrero.,
2015).

29
CHAPTER THREE: Optimization of the effect of formulation on nutritional quality, texture and
physicochemical properties of banana-vegetable soup powder

Abstract

There are reported cases of malnutrition in developing countries due to low levels of nutrients in staple
diets consumed in homes. Uganda being a developing country has been a victim of such cases. The
largest composition of the population consume cooking bananas at an annual rate of 220 kg/person
which is among the world‟s largest consumption rate. Uganda being a large consumer of cooking
bananas is also among the largest producers of the fruit. The availability of the fruit with little being
sent to the international market due to low shelf life and high post-harvest losses has initiated value
addition to the fruit. This has seen several products developing from cooking bananas, among which
include raw tooke flour for use in confectionaries, tooke porridges, snacks and cakes.In this research, a
nutrient rich soup flour is developed from banana flour by enriching it with amaranth flour, pumpkins,
tomatoes, mushrooms and carrots.……The purpose for this study was to optimizethe effect of
formulation of banana-amaranth and vegetables mixon texture,nutritional and physicochemical
properties of the soup. Response surface methodology (RSM) was carried out using Design expert (Stat
Ease Software, Version 11.0.0.1) to give an optimal composition of the soup. The mixture design in
design expert gave an optimum product composition consisting of banana-amaranths, pumpkins,
carrots and mushrooms at 82%, 9%, 5% and 4% respectively. The optimal product had an energy
composition of 409.39kCal/100g, peak viscosity of 2631.41Cp while the holding viscosity, breakdown
viscosity, final viscosity, peak time, pasting temperature, carbohydrates, proteins and zinc contents of
1430.11Cp, 1209.57Cp, 2495.29Cp, 4.9 minutes, 78.41oC, 65.38%, 14.86% and 13.50g/100g
respectively.Mathematical models for predicting nutritional composition, viscosity, physicochemical
properties were significant (p<0.05). The coefficient of determination R2 ranged from 0.40 (Peak time)
to 0.99 (fat content). Fat content, Protein Content, moisture content, in vitro protein digestibility, Fiber
content and Zinc with R2 values of 0.99, 0.93, 0.93, 0.91, 0.9, and 0.9 respectively. The lack of fit test
was not significant (p<0.05).
Key words: Banana, Amaranth, vegetables, soup, optimization; response surface methodology

3.1.Introduction

Children malnutrition remains a big challenge in low income and middle-income countries (Black et
al., 2008). Nutritional improvement of staple foods has been explored to reduce these deficiencies in

30
recent product developments. Bananas are highly available on the Ugandan market with a potential for
utilization to provide for nutrition demands of Uganda‟s population. Grain amaranth (Amaranthus) is
considered as one of the fastest growing andhigh yielding cropswith a potential to contribute to the
nutritional availability in bananas(Akande et al., 2017; Tibagonnzeka, et al.,2014). It contains superior
quality protein but has marginal levels of leucine (an essential amino-acid) and is limited in vitamin A
(0 µg RAE/100 g or 2 IU/ 100 g) (USDA, 2014). This implies that complementing the mix with
vegetables may enhance the quality of banana based soups as is the goal of this study. Pumpkins are
considered as good sources of zinc and iron and so are the carrots. Mushrooms and tomatoes were
considered as good sources of vitamins A and C as well as good supplements for minerals(Manzi et al.,
2001).Amaranth flour, Pumpkins, mushrooms and carrots were used to enrich matookeflour in the
development of a nutrient enriched soup.

3.2.Methods and materials

3.2.1. Raw material selection for use in this study

The plantain used in this study were the cooking type also locally known as matooke, a Musa sp.
triploid acuminate genome group (AAA-EAHB). The species used in this study were chosen due to
their better nutritional properties than other banana species(Kawongolo, 2013; Nyombi et al., 2009). To
improve the protein content of banana flour, grain amaranth flour was added because it is reported to be
a rich source of proteins (Soriano-Garcíaet al., 2018). The vegetables were chosen basing on their
nutrient values; carrots are rich in beta carotene (124.28 mg/kg) (Bystrickáet al., 2015), pumpkins are
rich in zinc(Ravi, Lakshmi, & Ranjani, 2010b) while mushrooms are rich in most minerals(Manzi et
al., 1999).

3.2.2. Sample preparation

Green cooking bananas were peeled under cold water treated with 1 percent sodium metasulphite to
avoid browning and sliced to about 2 mm thickness. The slices were laid for drying in air oven driers at
Jakana foods ltd, Kawempe, Kampala Uganda. The slices were dried at 60°C for 6 hours in an air
ovendriers (SAVER TRAY DRIER, Model: R-5A). Fresh Mushrooms were purchased from Capital
shopper‟s supermarket and were dried for 4 hours at 60oC using the same driers at Jakana. Carrots were
purchased from Kalerwe market in Kampala, chopped, sliced and laid on steel trays to dry for 12 hours
at 50oC in air oven driers(Pandey & Kulshrestha, 2003b). Pumpkins, and tomatoes were purchased

31
from Kalerwe market, Kampala transported in nylon bags chopped, sliced and laid on trays for drying
for 6 hours at 60oC. The dried samples were milled using a wonder mill. The flour was sieved using a
laboratory test sieve (Wagtech international, Ltd., 300 MIC, BS 410), packed in polythene papers, and
kept in plastic buckets for further analysis.
3.2.3 Experimental Design

The proportions of vegetables and banana powder used in this study were determined basing on the
nutritional requirements of children aged 6-59 months. The ingredients were entered into the Nutri-
Survey for windows (SEMEO-TROPMED RCCN, University of Indonesia) and adjustments in the
proportions of the ingredients made to meet the target percentages of proteins (16 g), energy (1060
kCal). The resultant ratios were used in the Design expert (Stat Ease, Version 11.1.0.1 Minneapolis) to
obtain runs for use during optimization process. Thirteen formulations were generated and presented in
table 3.1.

Table 3.1: Proportions of bananas and selected vegetables obtained using Nutri-survey
Formulation Amounts of ingredients g per 100 g of flour
code
Banana Grain Amaranths Pumpkins Tomatoes Mushrooms Carrots

S0 (Control) 50 50 0 0 0 0
S1 42.5 42.5 15 0 0 0
S2 42.5 42.5 0 15 0 0
S3 42.5 42.5 0 0 15 0
S4 42.5 42.5 0 0 0 15
S5 40 40 20 0 0 0
S6 40 40 0 20 0 0
S7 40 40 0 0 20 0
S8 40 40 0 0 0 20
S9 45 45 10 0 0 0
S10 45 45 0 10 0 0
S11 45 45 0 0 10 0
S12 45 45 0 0 0 10

32
S0-12 refers to samples with respective Banana, Amaranths, Pumpkins, Tomatoes, Mushrooms and
Carrot combinations.
3.2.4 Determination of nutritional properties

The moisture, crude protein, ash, total fat, minerals (iron, potassium and zinc), total carbohydrate,
dietary fiber, total fat and gross energy contents of the banana-based flours were determined using
standard methods.
3.2.4.1 Determination of moisture content

The moisture content of the samples was determined by oven drying overnight at 105°C (AOAC,
1999). Dried aluminum dishes were weighed and their respective weights recorded as W 1. About 2 g of
the ground sample was weighed using an analytical electronic balance into already weighed dishes
(W2) and dried in a fan oven at 105 °C overnight. After evaporation of moisture from the sample, it was
left to cool in a desiccator and weighed again to obtain weight after drying (W3). The experiment was
carried out in triplicate. The percentage average difference in the weights of three samples of flour,
before and after oven drying was calculated to give the moisture content.
𝐿𝑜𝑠𝑠 𝑖𝑛 𝑤𝑒𝑖𝑔𝑕𝑡
% 𝑚𝑜𝑖𝑠𝑡𝑢𝑟𝑒 𝑐𝑜𝑛𝑡𝑒𝑛𝑡 = 𝑥 100
𝑊𝑒𝑖𝑔𝑕𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒

3.2.4.2 Determination of crude protein content

Protein determination was carried out using the method as described by AOAC, (1990). The
determination of protein by micro Kjeldahl method involves three stages, the digestion stage, the
distillation stage and the titration stage.Weighed sample (1.0g) was digested with concentrated
sulphuric acid (20 ml) and digestion mixture (10.0 g) in Kjeldahl digestion flask. The contents were
cooled and transferred to 250 ml volumetric flask. The volume was made up to the mark with distilled
water and mixed. Measured aliquot (5 ml) was poured in distillation flask followed by 40% sodium
hydroxide and ammonium borate was collected through a condenser in a flask containing 10 ml of
4.0% boric acid solution. The distillate was titrated with 0.1 N sulphuric acid. A blank sample was also
run along with the samples(Bhat & Bhat, 2013).

𝑇𝑖𝑡𝑟𝑒𝑣𝑎𝑙𝑢𝑒𝑥 0.0014 𝑥 volume made

% 𝑵𝒊𝒕𝒓𝒐𝒈𝒆𝒏 = 𝑋100
Aliquote taken g 𝑥 Weight of sample (g)

% 𝑃𝑟𝑜𝑡𝑒𝑖𝑛 = % 𝑁𝑖𝑡𝑟𝑜𝑔𝑒𝑛 𝑥 6.25

33
Determination of fat content

Total fat content was determined using the Soxhlet method (AOAC, 1999). About 4g of the sample was
weighed into thimbles; 40 ml of extraction solvent (Petroleum Ether) was measured into beakers, the
thimbles containing the samples placed in the beaker containing the petroleum ether which was later
fixed on a Soxtec equipment. Fat extraction was done by boiling the samples for about an hour. The
solvent was distilled off and the fat dried in an air oven at 100 °C for about 30 minutes. The oil
collected in the beaker was weighed. Percentage fat content was determined using the following
formula:
𝑊3 − 𝑊2
𝐹𝑎𝑡 𝑐𝑜𝑛𝑡𝑒𝑛𝑡 = 𝑋 100
𝑊1
Where: W1 = weight of sample, W2 = weight of empty beaker, W3 = weight of beaker + fat

3.2.4.3. Determination of Ash content

The ash content was determined as described by AOAC, (1990). The crucibles for ashing were properly
washed and dried in a hot air oven and allowed to cool in desiccators. The crucibles were weighed and
their weight recorded as W1. About 2g of the flour sample (W2) was weighed into the crucible and the
weight noted (W3). The crucibles and their content were transferred into the muffle furnace set at and
maintained at 550°C before timing commences for six hours. The ashing was complete when there is
no black speck in the ash that is the sample turned grey. The crucibles containing the ash were taken
out and transferred to the desiccators to cool after which the weights were taken. The difference in
weight between the weight of the crucible and the crucible plus the ash was calculated as ash content.
The percentage of ash was calculated as follows:
𝑊𝑒𝑖𝑔𝑕𝑡 𝑜𝑓 𝑎𝑠𝑕
% 𝐴𝑠𝑕 𝑐𝑜𝑛𝑡𝑒𝑛𝑡 = 𝑥 100
𝑊𝑒𝑖𝑔𝑕𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒

3.2.4.4. Dietary fiber content

The dietary fiber content of the mixed flours was determined using the FOSS Fibertec 2010. About
1.0g of the sample was weighed into the crucible. The mixture for dietary fiber determination was
prepared from 20g of cetyltrimethylammonium bromide, concentrated sulphuric acid (28 ml) and
400ml of distilled water and made up to 1L with distilled water. The glass crucibles containing the
samples were fixed into the Fibertec machine. The temperature was set at 50°C and the mixture boiled

34
for 45minutes after which it was washed severally with distilled water. The glass crucibles were taken
to the oven maintained 100oC for 45 minutes to drive off the moisture. Dietary fiber was obtained as
the difference between the weight of the empty glass crucible and that after removal from the oven.
𝐿𝑜𝑠𝑠 𝑖𝑛 𝑤𝑒𝑖𝑔𝑕𝑡
% 𝐶𝑟𝑢𝑑𝑒 𝑓𝑖𝑏𝑟𝑒 = 𝑥100
𝑊𝑒𝑖𝑔𝑕𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒

3.2.4.5. Determination of total carbohydrates content

The carbohydrate content was determined by difference on dry weight basis. The total percentages of
the fat, crude protein, ash, dietary fiber was deducted from 100%, giving the amount of nitrogen-free
extract otherwise known as carbohydrate.

On dry weight basis

% CHO = 100 % − (% Fat + % Ash + % dietary Fibre + % Crude Protein)

3.2.4.6. In vitro protein digestibility

In vitro protein digestibility (IVPD) was determined according to the method by Khalil et al., 1984.
Approximately 0.2 grams of the sample was weighed into two 50 milliliter centrifuge tubes. To
determine digestibility, two milliliters of distilled water was added to the 0.2g samples, shaken and the
tubes was placed in a boiling water bath for 20 minutes. To each of the sample, 20 milliliters of 0.1 M
phosphate buffer with pepsin (1:3000 IU Hog pepsin/L, pH 2.0) was added. A blank was prepared in a
similar manner without the sample. The tubes were incubated in a shaking water bath at 37 oC, for two
hours and then centrifuged at 6000 rpm for 15 minutes. The supernatant was removed with a dropper
and discarded. Ten milliliters of Phosphate buffer were added to each tube and centrifuged at 6000 rpm
and the supernatant discarded. The residue was removed and placed in the center of a filter paper on a
Buchner funnel. Suction was applied to the filter flask and the remaining residue rinsed from the tube
into the funnel using five milliliters of the buffer. The filter papers were rolled and inserted into
Kjeldahl flasks. The flasks were dried in the oven at 100 oC for a minimum of 15 minutes. Into the
Kjeldahl flasks containing the filter paper and sample, ten milliliters of concentrated Sulphuric acid,
one gram of Potassium Sulphate and one milliliter of ten percent Copper Sulphate solution was added.
Digestion, distillation, and titration was done as described in the determination of initial protein
content. Percentage protein digestibility was calculated as follows:
A−B
% In vitro protein digestibility = A

35
Where;
A = % Protein in sample before digestion
B = % Protein in sample after pepsin digestion

3.2.4.7. Determination of gross energy content

Gross energy content was determined by oxygen bomb calorimeter (GallenkampAutobomb) (AOAC,
1999). About 1 g of the sample was weighed, pelleted and placed in a clean combustion crucible. A
platinum wire (10cm) was attached between the electrodes of the bomb. The combustion crucible
containing the sample was set in place in the loop electrode. A cotton thread was tied in the middle of
the platinum wire. The thread was adjusted until it touches the sample. Distilled water (200 ml) was put
in the calorimeter bucket and placed in the calorimeter. The bomb was filled with oxygen to 30
atmospheres gauge pressure. The cover of the calorimeter was closed, the thermometers lowered and
the circulating motor was started. The temperature of the water in the outer jacket was adjusted to equal
that of the calorimeter. The initial temperature of the calorimeter was recorded as Ti. The sample was
then ignited. The final temperature was read and recorded as Tf. The calorimeter was then opened and
the bomb was taken out of the bucket. The acid correction was done by rinsing all inner bomb surfaces
with distilled water and collecting all the washings in a clean beaker. The washings were titrated
against 0.1 N sulphuric acid with methyl orange indicator. Gross energy was calculated using the
following formula:
𝐺𝑟𝑜𝑠𝑠 𝑒𝑛𝑒𝑟𝑔𝑦 (𝐾𝑐𝑎𝑙 𝑝𝑒𝑟 𝑔 𝐷𝑀) = ((𝑇𝑓 − 𝑇𝑖) − (𝑐𝑜𝑟𝑟𝑒𝑐𝑡𝑖𝑜𝑛 𝑓𝑎𝑐𝑡𝑜𝑟) 𝑥 𝐻𝑒)/(𝑊 𝑥 𝐷𝑀%)
Where: Tf = final temperature, Ti = initial temperature, W = weight of sample, He = hydrothermal
equivalent, correction factor = w + titer, titer = amount of sulphuric acid used, w = length of wire left.

3.2.5 Determination of Physicochemical properties of the flours

3.2.5.1 Bulk density

A 50-gram sample of flour was poured from a constant height into an empty pre-weighed 100
milliliters measuring cylinder up to a certain height. The cylinder was tapped continuously and
carefully until no further decrease in the level of flour is observed at the graduation mark and the
weight of the cylinder taken. Bulk density was expressed as grams per milliliters. For each formulation,
three replicates were made.

36
3.2.5.2 Water absorption capacity

The water absorption capacity (WAC) of flours was measured using the centrifugation method reported
by (Kaur and Singh, 2006). Each sample of three grams was dissolved in 25 milliliters of distilled water
and placed in a 50 mL pre-weighed centrifuge tube. The mixture was stirred at five minutes interval,
held for 30 minutes, followed by centrifugation for 30 minutes at 3,000×g. The supernatant was
decanted, excess moisture removed by drying at 50 oC for 25 minutes and then the sample was
reweighed.
3.2.4.8. Water absorption index and water solubility index

The water absorption index (WAI) and water solubility index (WSI) of samples was determined by
methods reported by Kaur and Singh, 2006. A sample of three grams was dissolved in 30 milliliters of
distilled water and heated in a water bath at 90 oC for 15 minutes. The cooked paste was cooled to room
temperature, transferred to pre-weighed centrifuge tubes, and centrifuged at 3,000×g for ten minutes.
The supernatant was decanted into a pre-weighed evaporating dish to determine its solid content and
the sediment was weighed. The weight of dry solids was recovered by evaporating the supernatant
overnight at 105 oC. The WAI and WSI was calculated as follows:
Weight of sediment
WAI =
Weight of flour sample
Weight of dissolved solids in supernatant
WSI =
Weight of flour sample

Pasting properties of soups

Pasting properties of instant soups were determined using a Rapid Visco Analyzer (RVA-4, Newport
Scientific). The pasting temperature (PT), peak viscosity (PV, the maximum hot paste viscosity),
holding strength or trough viscosity (the trough at the minimum hot paste viscosity), final viscosity,
breakdown (BD, peak viscosity-holding strength or trough viscosity) and setback (SB, final viscosity-
holding strength) were obtained using Thermocline for Windows software. The viscosities were
presented in Centipoises. The pasting time were presented in minutes.
3.2.6 Sensory evaluation of formulations for acceptability of the formulations.

Sensory evaluation of samples was carried out to choose the most acceptable formulation. To improve
on sensory acceptability of the soup, the samples were prepared using chicken broth prepared from
chicken legs, parsley, thyme, bay leaves, onions, garlic, salt and water. The samples were prepared by

37
mixing 50g of the samples together with 800mls of chicken broth, boiled and allowed to simmer for
five minutes. The prepared samples were served to 30 panelists from the school of food science and
technology. The most acceptable combination of flours basing on the 9-point hedonic scale were
chosen.

3.2.7 Data analysis

All experiments were conducted in triplicate. Statistical analysis of the data was performed by analysis
of variance (ANOVA), using SPSS software (IBM statistics, version 20). A probability value of
difference p ≤ 0.05 was considered to denote statistical significance. All data are presented as mean
values ± standard deviation (SD). Regression analysis was performed to indicate the relationship
between the tested parameters and formulations.

3.3 Results

3.3.1 Effect of variation of ,,,,,,,…formulations on the Nutritional composition of the soups

The formulations presented different values for the tested parameters.Each of the formulation has 50%
Banana, 50% Amaranths grains mixed with any of 10%, 15%, and 20% for pumpkins, tomatoes,
mushrooms and carrots. The samples were formulated and different tests carried whileoptimizing the
ingredients of the formulation. Results from proximate composition of the formulations are presented
in table 3.2.

5
4 80
3 60
2 40
1 20
0 0

SC SC SC SC
FC FC FC FC

20 80
15
10 60
5
0 40

20

0
Pumpkin Tomato Mushrooms Carrots
38 Carbohydrate Carbohydrate
PC PC PC PC Carbohydrate Carbohydrate
 Figure 3.1: Variation of nutrition values in the formulation;Fat content (FC),
Protein content (PC), Carbohydrates and Starch content (SC).

The gross energy content (GE) varies between 369.41±11.16 and 441.98±5.17kCal/100g. The
formulation with pumpkin at 10%, 15% and 20% have a GE of 390.53, 410.50 and 411.31 respectively,
The formulation with tomatoes at 10%, 15% and 20% have a GE of 410.59, 386.93 and 414.07
kCal/100g respectively.The formulation with mushrooms in the order of 10%, 15% and 20% have a GE
of 416.38, 386.93, 391.69 kCal/100g respectively. The formulation with carrots at 10%, 15% and 20%
have a GE of 414.19, 441.98, and 401. 55 kCal/100g respectively. However, the formulation with 15%
pumpkins, 20% pumpkins and 10% carrots are not significantly (p=0.05) different from each other.The
IVPD content varies between 58.90 and 72.71%. The formulation with pumpkin at 10%, 15% and 20%
have an IVPD content of 72.56, 66.70 and 71.16% respectively. The formulation with tomatoes at 10%,
15% and 20% have IVPD composition of 71.73, 65.09 and 66.27% respectively. The formulation with
mushrooms in the order of 10%, 15% and 20% have an IVPD composition of 58.90, 71.70 and 62.63%
respectively. The formulation with carrots at 10%, 15% and 20% have an IVPD content of 67.35, 69.88
and 72.71% respectively. The addition of carrots improved the IVPD content relative to the control.
There is no statistical significance between variations of IVPD in the formulations. The fat content of
the formulations vary between 2.61±0.10 and 4.01±0.10 %. The formulation with pumpkin at 10%,
15% and 20% have a fat content of 2.72, 2.91, and 3.71 respectively. The formulation with tomatoes at
10%, 15% and 20% have a fat content of 2.90, 3.26 and 3.83 respectively. The formulation with
mushrooms in the order of 10%, 15% and 20% have a fat content of 3.03, 3.27 and 4.01 % respectively.
The formulation with carrots at 10%, 15% and 20% have a fat content of 2.61, 2.64 and 3.88%
respectively. However, the formulation with 10% composition of pumpkins, tomatoes, mushrooms and
carrots are not significantly different from each other. The protein content of the formulations varies
between 12.00±0.65 and 17.20±0.53%. The formulation with pumpkin at 10%, 15% and 20% have a
protein content of 12.85, 13.79, and 15.29% respectively. The formulation with tomatoes at 10%, 15%
and 20% have a protein content of 12.99, 13.51 and 15.27% respectively. The formulation with
mushrooms in the order of 10%, 15% and 20% have a protein content of 14.56, 15.50 and 17.20 %

39
respectively. The formulation with carrots at 10%, 15% and 20% have a protein content of 12.25, 12.92
and 14.29% respectively. However, the formulation with 10% carrots and the control are not
significantly different from each other.The carbohydrates of the formulations were determined by a
method of difference. The carbohydrates content of the formulations varies between 63.02 and 70.67%.
The formulation with pumpkin at 10%, 15% and 20% have a carbohydrates content of 68.46, 66.64 and
64.76% respectively.The formulation with tomatoes at 10%, 15% and 20% have a carbohydrates
content of 66.79, 65.55 and 63.33% respectively. The formulation with mushrooms in the order of
10%, 15% and 20% have a carbohydrates content of 68.93, 65.71 and 63.02% respectively. The
formulation with carrots at 10%, 15% and 20% have a carbohydrates content of 68.76, 68.57, and
65.00% respectively.The starch content of the formulations varies between 55.83±0.56 and
68.52±0.74%. The formulation with pumpkin at 10%, 15% and 20% have a starch content of 63.38,
60.18 and 60.33% respectively. The addition of pumpkins thus reduces starch content in the
formulations. The formulation with tomatoes at 10%, 15% and 20% have a starch content of 59.13,
55.83 and 56.70% respectively. The formulation with mushrooms in the order of 10%, 15% and 20%
have a starch content of 63.35, 64.03 and 61.31% respectively. The formulation with carrots at 10%,
15% and 20% have a starch content of 58.11, 57.63 and 57.37% respectively, thus the change in starch
content due to carrots is in the order of Control>10%>15%>20%.

40
 Table 3.2: Proximate composition of different flour formulations for making soups
Formul MC (% ) Fat (%) Fiber (%) Protein (%) % Ash Carbohyd Starch
GE
ation rates (%) content
(kCal/100g) (g/100g)
Code* IVPD (%)

S0 414.75±0.21cd 9.69±0.19a 3.25±0.12bc 1.45±0.14a 12.00±0.65a 67.20±9.97a 2.94±0.02a 70.67 68.52±0.74g

S1 410.50±5.53bcd 10.96±0.29bc 2.91±0.13a 2.59±0.04bcd 13.79±0.18bc 66.70±5.99a 3.11±0.03bc 66.64 60.18±2.10de

S2 386.93±4.48ab 11.09±0.64c 3.26±0.08ab 3.34±0.39f 13.51±0.67bc 65.09±10.00a 3.25±0.07de 65.55 55.83±0.56a

S3 369.41±11.16a 9.56±0.27a 3.27±0.13ab 2.90±0.11bcde 15.50±0.70e 71.70±5.94a 3.06±0.04b 65.71 64.03±1.10f

S4 441.98±5.17e 10.02±0.24ab 2.64±0.80a 2.43±0.36bc 12.92±0.64ab 69.88±7.16a 3.42±0.04f 68.57 57.63±1.30abc

S5 411.31±11.28bcd 10.52±0.19abc 3.71±0.06bc 2.79±0.10bcd 15.29±0.90de 71.16±7.29a 2.93±0.02a 64.76 60.33±0.61de

S6 414.07±1.15cd 11.24±0.36c 3.83±0.05bc 3.02±0.17def 15.27±0.64de 66.27±1.54a 3.31±0.02e 63.33 56.70±0.41ab

S7 391.69±3.13abcd 9.66±0.47a 4.01±0.10c 3.17±0.18ef 17.20±0.53f 62.63±5.14a 2.94±0.04a 63.02 61.31±1.79e

S8 401.55±6.62bcd 10.68±0.42bc 3.88±0.19bc 2.95±0.05bc 14.29±0.64bc 72.71±5.61a 3.20±0.09cd 65.00 57.37±1.30abc

S9 390.53±4.77abc 10.67±0.25bc 2.72±0.05a 2.25±0.18b 12.85±0.84ab 72.56±8.44a 3.05±0.02b 68.46 63.38±0.99f

S10 410.59±0.35bcd 11.15±0.43c 2.90±0.03a 2.54±0.23bcd 12.99±0.35ab 71.73±7.94a 3.63±0.02g 66.79 59.13±0.74cd

S11 416.38±7.86d 9.67±0.26a 3.03±0.04a 3.81±0.04g 14.56±0.65cbd 58.90±9.84a 3.03±0.05g 68.93 63.35±1.10f

S12 414.19±7.48cd 10.43±0.23abc 2.78±0.10a 2.61±0.10bcd 12.25±0.62a 67.35±10.15a 3.17±0.03cd 68.76 58.11±2.11bc

Values represent mean ± standard deviation (n=3). Means in the same column with different superscripts are significantly different (p <
0.05). *Formulation codes represent samples in the respective formulations indicated in table 4-1.

40
Effect of formulations on micro nutrients composition of the soups

The formulations presented different values for the tested parameters. Each of the formulation has 50%
Banana, 50% Amaranths grains mixed with any of 10%, 15%, and 20% for pumpkins, tomatoes,
mushrooms and carrots. The samples were formulated and different tests carried on to optimize the
ingredients of the formulation. Results from nutrient composition of the formulations are presented in
table 3.3.

Table 3.3. Micronutrients compositions of the samples

Formulation Iron Zinc K Vit C Vit A RAE
code* (mg/100g) (mg/100g) (mg/100g) (mg/100g) (mg/100g)

S0 27.3 9.93 1115.2 10.61±2.34a 0.017±0.00a

S1 29.4 12.74 1017.2 14.25±1.04a 0.064±0.00a

S2 27.1 9.8 1098.4 24.52±2.67a 0.437±0.02d

S3 26.96 19.64 1134.4 16.19±1.25a 0.020±0.00a

S4 39.3 12.52 1188.8 16.64±1.14a 0.157±0.00b

S5 38.4 12.26 923.4 15.84±2.35a 0.062±0.00a

S6 42 13.28 1660.4 27.69±2.18a 0.449±0.02d

S7 45.8 16.81 1233.2 18.41±2.80a 0.014±0.00a

S8 38.1 13.92 1092 11.95±1.37a 0.205±0.02b

S9 21.4 8.13 1211.6 28.47±39.61a 0.063±0.00a

S10 26.2 10.76 1366 23.71±6.76a 0.321±0.14c

S11 32.7 15.98 1146.4 14.14±1.20a 0.019±0.00a

S12 16 9.09 1123.6 12.27±1.13a 0.167±0.01b

Values represent mean ± standard deviation (n=3). *Formulation code represent samples in the
respective formulations as indicated in table 4-1.

41
 50 25
40
20
30
20 15

10 10
0
5

0
Pumpkin Tomato Mushrooms Carrots
Zinc Zinc Zinc Zinc
Iron Iron Iron Iron

2000 35
1800 30
1600
1400 25
1200 20
1000
800 15
600
10
400
200 5
0
0
Pumpkin Tomato Mushrooms Carrots
Pumpkin Tomato Mushrooms Carrots
Potassium Potassium Potassium Potassium Vit. C Vit. C Vit. C Vit. C

Figure 3.2: Variation in mineral composition and micronutrients; Iron content, Zinc content,
Potassium content and Vitamin C in the formulations
The mineral and micronutrient composition of the soups formulations has been illustrated in figure 3.2
above. Iron content varies between 16 and 45.8 mg/100g. The formulation with pumpkin at 10%, 15%
and 20% have an iron content of 21.4, 29.4 and 38.4 mg/100g respectively. The formulation with
mushrooms in the order of 10%, 15% and 20% have an iron composition of 32.7, 26.96 and 45.8
mg/100g respectively. The formulation with carrots at 10%, 15% and 20% have an iron content of 16,
39.3 and 38.1 mg/100g respectively. The addition of vegetables improved iron content. Zinc varies
between 8.13 and 19.64 mg/100g. The formulation with pumpkin at 10%, 15% and 20% have a zinc

42
content of 8.13, 12.74 and 12.26 mg/100g respectively. The formulation with tomatoes at 10%, 15%
and 20% have a zinc composition of 10.13, 9.8 and 13.28 mg/100g respectively. The formulation with
mushrooms in the order of 10%, 15% and 20% have a zinc composition of 15.98, 19.64 and 16.81
mg/100g respectively. The formulation with carrots at 10%, 15% and 20% have a zinc content of 9.09,
12.52 and 13.92 mg/100g respectively. The Vitamin C content varies between 10.61 and 28.47
mg/100g. The control sample has a Vitamin C content of 10.61 mg/100g. The formulation with
pumpkin at 10%, 15% and 20% have a Vitamin C content of 28.47, 14.25 and 15.84 mg/100g
respectively.

Effect of formulations on physicochemical properties of the soups

The formulations presented different values for the tested parameters. Each of the formulation has 50%
Banana, 50% Amaranths grains mixed with any of 10%, 15%, and 20% for pumpkins, tomatoes,
mushrooms and carrots. The samples were formulated and different tests carried on to optimize the
ingredients of the formulation. Results from physico-chemical composition are presented in table 3.4.

Table 3.4: Physicochemical properties

Formulation FC (%) FS (%)
-3
code* Bulk density (kgm )

S0 774.03±9.81ef 3.55±0.33 ND

S1 813.57±19.99f 4.98±0.09 1.25±0.02

S2 705.00±16.55cd 1.27±0.02 0.84±0.73

S3 754.73±1.55de 1.22±0.03 ND

S4 645.40±0.87ab 5.07±0.19 ND

S5 753.93±15.44de 2.45±0.06 ND

S6 710.63±5.35cd ND ND

S7 681.67±0.61bc 3.74±1.39 ND

S8 682.30±5.20bc 6.08±0.26 3.65±0.15

S9 810.33±2.49f 1.25±0.05 ND

S10 769.70±1.39ef ND ND

43
 S11 725.10±2.95cde 5.54±0.84 ND

S12 631.37±59.93a 2.51±0.14 ND

Values represent mean ± standard deviation (n=3). Means in the same column with different
superscripts are significantly different (p < 0.05). *Formulation codes represent samples in the
respective formulations indicated in table 4-1.

1000
800
600
400
200
0

ρb ρb ρb ρb

7
4
6
3.5
5 3
4 2.5
3 2
2 1.5
1
1
0.5
0
0
Pumpkin Tomato Mushrooms Carrots
Pumpkin Tomato Mushrooms Carrots
FC FC FC FC FS FS

Figure 3.3: Variation of physicochemical properties in the formulation; bulk density (ρb),
Foaming Capacity (FC) and foaming stability (FS)
The variations in the physicochemical composition of the soups has been illustrated in figure 3.3 above.
Bulk density content varies between 631.37±59.93 and 813.57±19.99 kgm-3. The control sample has a
bulk density of 774.03 kgm-3. The formulation with pumpkin at 10%, 15% and 20% have a bulk

44
density content of 810.33, 813.57 and 753.93 kgm-3 respectively. The formulation with tomatoes at
10%, 15% and 20% have bulk density of 769.70, 705.00 and 710.63 kgm-3 respectively. The
formulation with mushrooms in the order of 10%, 15% and 20% have a bulk density of 725.10, 754.73
and 681.67 kgm-3 respectively. The formulation with carrots at 10%, 15% and 20% have a bulk density
of 631.37, 645.40 and 682.30 kgm-3 respectively. There isstatistical significance between variations of
bulk density in the formulations. The formulations could foam in some cases and not stable at some
cases. The foaming capacity varies between 1.22±0.03 and 6.08±0.26% while foaming stability for the
formulations is highly un predictable. The control sample has a foaming capacity of 3.55±0.33% with
no stability. The formulation with pumpkin at 10%, 15% and 20% have a foaming capacity of 1.25,
4.98 and 2.54% respectively. The only stable formulation is one where pumpkin is at 15%. The
formulation with tomatoesat 15% is at 1.27% with a stability of 0.84% while the foaming capacity and
stability of tomatoes at 10% and 20% are not defined.

Effect of formulations on pasting properties of the soups

Pasting properties of the banana-vegetables soups presented in Table 4-3 were obtained using standard
procedures presented byShimelis, et al.,(2006). A rapid Visco Analyzer (Model: RVA-4, New Port
scientific, Pty. Ltd., Australia) was used to evaluate testing properties of banana flours. 3.5g of flour
was weighed and put into a canister from which 25 mls of distilled water was added (Cheok et al.,
2018). The canister was inserted into the pasting cell and the run was started.The test runs included 1
minute of mixing, stirring and warming up to 50oC, 3 minutes and 42 seconds of heating at 12oC/min
up to 95oC, 2.5 minutes of holding at 95oC, 3 minutes and 42 seconds for cooling back to 50oC at the
same rate as heating and 2 minutes holding at 50oC. The process ends after 13 minutes.Significant
differences were observed in terms of pasting properties fordifferent formulations against control
formulation. The control formulation had the highest significant (p<0.05) peak viscosity, holding
viscosity and final viscosity compared to the rest of the formulations and significantly lower for
breakdown viscosity, setback viscosity, peak time and pasting temperature.

45
 4000 2000

Holding Viscosity
Peak Viscosity 3500 1800
3000 1600
2500 1400
2000 1200
1500 1000

Composition Composition
Pumpkin Tomato
Pumpkin Tomato
Mushrooms Carrots
Mushrooms Carrots

3500 1500
Final Viscosity

3000
Setback Viscosity
2500 1000
2000
1500
1000 500
0
Control 10% 15% 20%
Composition Composition
Pumpkin Tomato
Pumpkin Tomato
Mushrooms Carrots
Mushrooms Carrots

5.5 80
Peak Viscosity

Pasting temperature

78
5
76
4.5 74
4
Control 10% 15% 20%
Composition Composition
Pumpkin Tomato Pumpkin Tomato
Mushrooms Carrots Mushrooms Carrots

46
Figure 3.4: Pasting properties of the soup samples; Peak Viscosity, holding viscosity, final
viscosity, setback viscosity, peak time and pasting temperature.
General pasting properties of soup flour samples were determined using the RVA technology at the
department of food technology and nutrition of Makerere University. The results are vary as shown in
figure 3.4 above. RVA results indicate that the control formulation has the highest viscosities (final
viscosity, peak viscosity, holding viscosity, and setback viscosity) compared to other formulations and
that as the percentage composition of pumpkins, tomatoes, mushrooms and carrots increase, there is a
general decline in viscosities. In all the formulations, the peak viscosity lies in the range of 2338.5 to
3413.5 Cp, Holding Viscosity lies between 1212 to 1896.5 Cp, Breakdown Viscosity lies between
842.0 to 1628 Cp. The final Viscosity lies between 766.5 to 1276.5 Cp, the peak time lies between 4.63
to 5.17 minutes while the pasting temperature lies between 76.28 to 79.53 oC.General pasting
properties have been indicated in table 3.5 below.

47
 Table 3.5: Pasting properties of flour samples
Formulation Peak Viscosity Holding Breakdown Final Viscosity Setback Viscosity Peak Time Pasting
1 2
code* (Cp) viscosity (Cp) Viscosity (Cp) (Cp) (Cp) (Minutes) Temp. (oC)
S0 3413.50±45.96g 1896.50±51.62h 1517.00±97.58cde 2925.50±12.02e 1029.00±63.64cd 4.73±0.09abc 78.70±0.57cde
S1 2952.00±26.87cdef 1605.50±17.68ef 1346.50±44.55bcde 2771.00±9.90de 1165.50±7.78cde 4.83±0.05abcd 78.75±0.49de
S2 3336.00±288.5fg 1654.50±23.33fg 1681.50±265.17e 2858.50±200.11e 1204.00±176.78de 4.77±0.14abcd 76.33±0.53a
S3 2865.50±10.61cde 1473.00±38.18cd 1392.50±27.58cde 2239.50±2.12ab 766.50±36.06ab 4.67±0.00ab 77.43±0.04abcd
S4 2338.50±12.02ab 1313.50±10.61ab 1025.00±1.41ab 2530.50±17.68cd 1217.00±7.07de 5.00±0.00de 78.70±0.57cde
S5 2817.00±14.14cde 1528.00±12.73de 1289.00±1.41bcd 2670.50±3.54de 1142.50±9.19cde 4.90±0.05bcd 78.33±0.04bcde
S6 2749.00±132.94cd 1492.50±4.95cde 1256.50±127.99bc 2520.00±91.92cd 1027.50±86.97cd 4.93±0.00cde 76.28±0.53a
S7 2646.00±28.28bc 1352.50±6.36b 1293.50±21.92bcd 2010.50±17.68a 658.00±11.31a 4.67±0.00ab 77.10±0.64abc
S8 2054.00±22.63a 1212.00±46.67a 842.00±24.04a 2288.50±19.09bc 1076.50±27.58cde 5.17±0.05e 79.53±0.53e
S9 3206.00±8.49efg 1746.00±15.56g 1460.00±7.07cde 2842.50±0.71e 1096.50±14.85cde 4.77±0.05abcd 78.30±0.14bcde
S10 3271.50±140.71fg 1643.50±54.45fg 1628.00±86.27de 2764.00±66.47de 1120.50±12.02cde 4.73±0.09abcd 76.70±0.00ab
S11 3075.00±0.00defg 1576.50±2.12def 1498.50±2.12cde 2530.50±10.61cd 954.00±8.49bc 4.63±0.05a 77.50±0.14abcd
S12 2641.00±9.90bc 1404.00±0.00bc 1237.00±9.90bc 2680.50±53.03de 1276.50±53.03e 4.93±0.00cde 79.15±0.00e
Values represent mean ± standard deviation (n=3). Means in the same column with different superscripts are significantly different (p <
0.05). *Formulation codes represent samples in the respective formulations indicated in table 4-1.1Break down Viscosity= Peak –
2
Holding Viscosity, Setback Viscosity=Final-Holding Viscosity.

47
2.9. Optimization of responses

Fitting the response surface models for Nutritional composition of the enriched banana vegetable
soup.

The „„fitness‟‟ of the models and their suitability to accurately predict the variation were studied using
coefficients of determination (R2) and F-values as shown in Table 3.6. All the model terms were
significant (p<0.05). The coefficient of determination R2 ranged from 0.40 (Peak time, pasting
property) to 0.99 (fat content). The R2 is the proportion of variability in the response values explained
or accounted for by the model. The closer the value of R2 to unity, the better the empirical models fit
the actual data and the more relevant the dependent variables in the model have in explaining the
behavior of variations. Responses whose coefficients of determination were so close to unity were
analyzed. These included Fat content, Protein Content, moisture content, Overall acceptability, in vitro
protein digestibility, Fiber content and Zinc with R2 values of 0.99, 0.93, 0.93, 0.91, 0.91, 0.9, and 0.9
respectively. The lack of fit test was not significant (p<0.05).
Model reduction was carried out by dropping the insignificant terms based on the p-value >0.05 in
order to improve the model equations using the automatic selection approach.

Table 3.6: Coefficients for the fitted first order and second-order polynomials representing the
relationship between the response and formulations of vegetable chicken soup mixture
Responses
GE MC Fat FC PC Vit C Carb IVPD Fe Zn
X1 41.49 0.1 0.03 0.02 0.11 0.09 0.71 0.74 0.27 0.1
X2 210.91 -0.46 0.8 0.08 0.21 1.79 -2.96 -0.18 9.25 1.52
X3 121.8 0.19 0.58 0.1 1.01 1.47 2.33 6.39 8.78 1.33
X4 25.65 -0.37 0.66 -1.03 0.44 1.55 2.76 -3.09 6.3 -2.14
X5 -93.11 -0.22 0.68 0.08 0.27 1.49 0.41 -2.4 10.02 2.1
X1X2 -2.08 0.01 -0.01 - - -0.02 0.04 - -0.11 -0.02
X1X3 -1.04 - -0.01 - -0.01 -0.02 -0.02 -0.07 -0.1 -0.01
X1X4 0 0.01 -0.01 0.01 - -0.02 -0.03 0.05 -0.07 0.03
X1X5 1.66 0 -0.01 - - -0.02 - 0.04 -0.11 -0.02
R2 0.57 0.93 0.99 0.9 0.93 0.54 0.82 0.91 0.75 0.9
Model
(F-value) 13.08 139.58 605.8 130.16 188.83 9.66 44.91 98.72 25.26 76.29
X1= Banana and Amaranth. X2= Pumpkins X3= Tomatoes X4= Mushrooms X5= Carrots, X1-5 predicts
single effects while X1X2, X1X3, X1X4, and X1X5 predicts interaction effects.

48
3.2.4.9. Model graphs

Figure 3.5: 3-D response surface plots showing the effect of ingredient mixes on Zinc content, IVPD, Fat and
protein Content.

49
3.2.4.10. Predictive models for nutrition composition of the formulations

Models predicting the variability of textural properties were developed in addition to models predicting
Variability of nutritional properties and physicochemical properties.A mathematical model presented in
equation 3.1 was built through regression based on the coefficient data in Table 3.6 for predicting the
variability of zinc in the formulation. Zinc content ranges between 8.13 and 19.64 mg/100g. The R 2 of
the second order polynomial model was good and explained 90% of the variability.

Zn  0.1X 1  1.52 X 2  1.33 X 3  2.14 X 4  2.10 X 5  0.02 X 1 X 2  0.01X 1 X 3  0.03 X 1 X 4  0.02 X 1 X 5 (R 2 = 0.90)
(3.1.)

The interaction and single variations were all significant terms of the model (p<0.05). The model
explains that changes in X2, X3 and X5 causes a significant changes in zinc content with X5 causing the
most significant change. The coefficients for the model terms shows high positive changes in the
model. The variations of pumpkin, carrots and tomatoes in figure 3.5 ranges between 0 and 12, with the
highest effects in the response identified when Pumpkins, carrots and tomatoes are at the maximum. It
can therefore be concluded that Zinc content of the soup increased with increase in the proportion of
carrots, pumpkin and tomatoes respectively in the formulation. The interaction factor for mushrooms
was negative implying that Zn reduces with increase in the proportion of mushrooms in the
formulation. The Quadratic interaction between Banana-Amaranth with Pumpkin, Tomatoes, Carrots
had a significant negative effect on Zn content while the interaction effect between Banana-Amaranth
and Mushrroms had a positive significant effect on Zinc content of the sample. It can be concluded that
mushrroms do not contribute significantly to the zinc content of the samples. The curve indicated in 3.5
indicates that the Quadratic effect was negative. The R2 of the predicted model was high and attained
high significance in relation to other models (R2 = 0.90, p = 0.0001). The model had a non significant
lack of fit which suggests a good fit to the mathematical model in Equation (3.1). This means that this
model is valid and can be used in subsequent prediction and optimisation stages.
A mathematical model in equation 3.2 for In vitro Protein digestibility (IVPD) of the sample was built
based on coefficient data in Table 3.6. Equation 4.2predicts variability of IVPD in the soup. IVPD
ranges between 55.7 and 77.6% in the samples. The R2 of the second order polynomial model was good
and explained 91% of the variability.
IVPD  0.74 X 1  0.18 X 2  6.39 X 3  3.09 X 4  2.40 X 5  0.07 X 1 X 3  0.05 X 1 X 4  0.04 X 1 X 5 (R 2 = 0.91)
(3.2.)

50
The interaction and single variations were all significant terms of the model (p<0.05). The model
explains that changes in X3 causes a significant positive change in IVPD while X4 produces a
significant negative change in IVPD. The variations of pumpkin, carrots and tomatoes from the curve
ranges between 0 and 12, with the highest effects in IVPD occurs when Pumpkins, carrots and
tomatoes are at the maximum. It can therefore be concluded that IVPD of the soup increased with
increase in the proportion of tomatoes in the formulation. The interaction factor for pumpkins,
mushrooms and carrots were negative implying that IVPD reduces with increase in the proportion of
pumpkins, mushrooms and carrots in the formulation. The Quadratic interaction between Banana-
Amaranth with Tomatoes had a significant negative effect on IVPD content while the interaction effect
between Banana-Amaranth with Mushrroms and carrots had a significant negative effect on IVPD of
the sample. It can be concluded that mushrooms, pumpkins and carrots do not contribute significantly
to the IVPD in the formulation. The curve indicated in figure 3.5 indicates that the Quadratic effect was
positive.
The R2 of the predicted model was high and attained high significance in relation to other models (R2 =
0.91, p = 0.0001). The model had a non significant lack of fit which suggests a good fit to the
mathematical model in Equation (3.2). This means that this model is valid and can be used in
subsequent prediction and optimisation stages.
A mathematical model in equation 3.3 predicts variability of Fat Content (Fat) content in the sample
based on the coefficient data in Table 3.6. The R2 of the second order polynomial model was good and
explained 99% of the variability.
Fat  0.03 X 1  0.80 X 2  0.58 X 3  0.66 X 4  0.68 X 5  0.01X 1 X 2  0.01X 1 X 3  0.01X 1 X 4  0.01X 1 X 5 (R 2 = 0.99)
(3.3.)

The interaction and single variations were all significant terms of the model (p<0.05). The model
explains that changes in X1, X2, X3, X4 and X5 causes a positive significant change in fat content with
X2 causing the most significant change. This indicates that fats in the sample are greatly contributed by
pumpkins in the sample. The variations of pumpkin, carrots, mushrooms and tomatoes from the curve
ranges between 0 and 12, with the highest effects in the response identified indicates identified when
Pumpkins, carrots and tomatoes are at the maximum. It can therefore be concluded that fat content of
the soup increased with increase in the proportion of carrots, mushrooms, pumpkin and tomatoes
respectively in the formulation. The Quadratic interaction between Banana-Amaranth with Pumpkin,
Tomatoes, Mushrooms and Carrots had a significant negative effect on Fat content. The curve indicated

51
in figure 3.5 indicates that the Quadratic effect was positive. The R2 of the predicted model was high
and attained the highest significance in relation to other models (R2 = 0.99, p = 0.0001). The model had
a non significant lack of fit which suggests a good fit to the mathematical model in Equation (3.4).
This means that this model is valid and can be used in subsequent prediction and optimisation stages.
A mathematical model in equation 3.4 for Protein Content (PC) content of the sample was built through
regression based on the coefficient data in Table 3.6. Equation 3.4 describes the relationship between
PC and soup formulation. PC content ranges between 11.26 and 18.01%. The R2 of the second order
polynomial model was good and explained 93% of the variability.
PC  0.11X 1  0.21X 2  1.01X 3  0.44 X 4  0.27 X 5  0.01X 1 X 3 (R 2 = 0.93) (3.4.)

The interaction and single variations represented were all significant terms of the model (p<0.05). The
model explains that changes in X2, X3 and X5 causes a significant changes in PC with X5 causing the
most significant change. The coefficients for the model terms shows high positive changes in the
model. The variations of pumpkin, carrots and tomatoes from the curve ranges between 0 and 12, with
the highest effects in the response identified indicates identified when Pumpkins, carrots and tomatoes
are at the maximum. It can therefore be concluded that PC of the soup increased with increase in the
proportion of carrots, pumpkin and tomatoes respectively in the formulation. The interaction factor for
mushrooms was negative implying that PC reduces with increase in the proportion of mushrooms in the
formulation. The R2 of the predicted model was high and attained high significance in relation to other
models (R2 = 0.93, p = 0.0001). The model had a non significant lack of fit which suggests a good fit
to the mathematical model in Equation (6). This means that this model is valid and can be used in
subsequent prediction and optimisation stages.
A mathematical model in equation 3.5 for Fibre Conctent (FC) content of the sample was built based
on the coefficient data in Table 3.6. Equation 3.5 describes the relationship between FC and soup
formulation. The R2 of the second order polynomial model was good and explained 90% of the
variability.
FC  0.02 X 1  0.08 X 2  0.1X 3  1.03 X 4  0.08 X 5  0.01X 1 X 4 (R 2 = 0.90) 3.5.

The model explains both single effects and interaction effects for X1X4 . The interaction and single
variations were all significant terms of the model (p<0.05). The model explains that changes in X2, X3
and X5 causes a significant changes in PC with X5 causing the most significant change. The coefficients
for the model terms shows high positive changes in the model. The variations of pumpkin, carrots and

52
tomatoes from the curve ranges between 0 and 12, with the highest effects in the response identified
indicates identified when Pumpkins, carrots and tomatoes are at the maximum. It can therefore be
concluded that PC of the soup increased with increase in the proportion of carrots, pumpkin and
tomatoes respectively in the formulation. The interaction factor for mushrooms was negative implying
that FC reduces with increase in the proportion of mushrooms in the formulation. The Quadratic
interaction between Banana-Amaranth with Pumpkin, Tomatoes, Carrots had a significant negative
effect on FC content while the interaction effect between Banana-Amaranth and Mushrroms had a
positive significant effect on FC of the sample. It can be concluded that mushrroms do not contribute
significantly to the FC of the samples. The curve indicated in figure 3.5 indicates that the Quadratic
effect was positive. The R2 of the predicted model was high and attained high significance in relation
to other models (R2 = 0.90, p = 0.0001). The model had a non significant lack of fit which suggests a
good fit to the mathematical model in Equation (3.5). This means that this model is valid and can be
used in subsequent prediction and optimisation stages.
3.2.4.11. Optimal solutions for selection of nutrient enriched soup flour

The Optimal formulation was obtained basing on the behavior of starch (pasting properties) and
nutritional composition of the formulation. The selection of the model to use was based on the value
with the greatest desirability using Desirability Function approach (DFA). The final composition of
flour with the best combination according to the criteria was 81.67% of bananas mixed with Amaranth
in the ratio of 1:1, 4% Mushrooms and 5.34% Carrots at a desirability index of 0.51. The formulation
had an energy composition of 409.39 kCal/100g), with peak, holding (hot paste), breakdown, final and
setback viscosities of 2631.41, 1430.11, 1209.57, 2495.29 and 1056.92 Cp respectively. The peak time,
Pasting Temperature, Carbohydrates, Protein content,IVPD and Zinc content are 4.9 minutes, 78.41 oC,
65.38%, 14.86%, 67.83%, and 13.5 respectively.

53
Table 3.7: Optimal solutions for nutritious and viscous soup formulations.
Number 1 2 3 4 5
Banana + Amaranths 81.67 80.00 81.58 80.00 80.00

Pumpkins (%) 9.24 10.41 12.53 12.89 13.74

Tomatoes (%) 0.00 0.00 0.19 0.85 0.00

Mushrooms (%) 3.76 3.62 0.00 0.00 0.00

Carrots (%) 5.34 5.97 5.71 6.26 6.26

GE (kCal/100g) 409.39 413.45 415.93 420.03 420.75

Peak Viscosity (cP) 2631.41 2557.80 2645.15 2574.98 2575.85

Holding viscosity (cP) 1430.11 1403.42 1459.55 1433.72 1434.81

Breakdown Viscosity (cP) 1209.57 1176.56 1195.13 1164.21 1164.36

Final Viscosity (cP) 2495.29 2468.71 2612.53 2579.33 2584.79

Setback Viscosity (cP) 1056.92 1043.98 1148.06 1132.38 1137.29

Peak Time (Minutes) 4.90 4.94 4.95 4.98 4.98

Pasting Temp. (oC) 78.41 78.45 78.66 78.60 78.69

Carbohydrates (%) 65.38 64.56 65.62 64.86 64.91

Protein content (%) 14.86 15.31 14.48 14.95 14.96

IVPD (%) 67.83 67.68 68.71 68.09 68.08

Zinc (g/100g) 13.50 13.80 12.38 12.96 13.00

Desirability 0.51 0.50 0.50 0.49 0.49

Selected

2.10. Discussion and Conclusion

The results indicated that there is a significant change (p=0.05) in texture as well as nutritional
composition and physico-chemical properties.The pasting properties of bananas-amaranth mixture
indicated the highest Viscosities due to the starch content in Bananas being higher (68.52%) as
indicated in table 2.1. Viscosities tend to go down as the starch content goes down and this is due to the
54
association of starch with other components such as proteins, fats among others(Cheok et al., 2018;
Shimelis et al., 2006). The selected formulation presented in table 3.7 has a gross Energy Content of
409.39 kCal/100g which is greater than the energy reported for bananas. The protein content of 14.86%
is higher than that reported by Muranga et al., (2010) of 5.57%. The peak Viscosity (219.28RVU),
holding viscosity (119.18 RVU), and breakdown viscosity (100.80 RVU) are lower than the pasting
properties exhibited by 100% banana flour by Muranga et al., (2010)of 375.92 RVU, 201.29 RVU and
174.63 RVU for peak, holding and breakdown viscosities respectively which are good properties for
use in the baking industries (Daramola & Osanyinlusi, 2006). This is explained by the reduction in the
starch content of flours when the starch rich banana flour is replaced with the vegetable flours (Adeniji
et al., 2010; Shimelis et al., 2006). The reduction in the viscosities brings this to the viscosity range for
children below five years of between 2000 – 3000 Cp (1Cp=12RVU). This indicates that the soup is
palatable to the target group. It can be concluded that the addition of vegetables in bananas improves
the nutrient composition of banana flour as well as the texture, physicochemical and mineral
composition of the samples thus allowing for the development of a good quality soup from banana-
vegetable mixes. Further developments to improve acceptability of the soup and also make the product
a user friendly product were adopted and agrees to extrusion as a legit technology for producing instant
products. Chicken being one of the flavors consumed by majority of people was incorporated into the
soup samples and its effect analyzed.

55
2.11. References

1. Adeniji, T. A., Hart, A. D., Tenkouano, A., Barimalaa, I. S., & Sanni, L. (2010). Comparative
study of the pasting properties of improved plantain. banana and Cassava Varieties with
Emphasis on Industrial application. African Journal of Food Agriculture Nutrition and
Development, 10(5), 210.
2. Akande, O. A., Nakimbugwe, D., & Mukisa, I. M. (2017). Optimization of extrusion conditions
for the production of instant grain amaranth-based porridge flour. Food Science and Nutrition,
5(6), 1205–1214. https://doi.org/10.1002/fsn3.513
3. AOAC. (1990). AOAC Official Methods of Analysis. Association of Official Agricultural
Chemists. Washington, D.C., 1(15th edition), 136–138.
4. Bhat, M. A., & Bhat, A. (2013). Study on Physico-Chemical Characteristics of Pumpkin
Blended Cake. Journal of Food Processing & Technology, 04(09).
https://doi.org/10.4172/2157-7110.1000262
5. Cheok, C. Y., Sulaiman, R., Manan, N. A., Zakora, A. J., Chin, N. L., & Hussain, N. (2018).
Pasting and physical properties of green banana flours and pastas. International Food Research
Journal, 25(6), 2585–2592.
6. Daramola, B., & Osanyinlusi, S. A. (2006). Production , characterization and application of
banana ( Musa spp ) flour in whole maize. African Journal of Biotechnology, 5(10), 992–995.
Retrieved from http://www.academicjournals.org/AJB
7. Kaur, M., & Singh, N. (2006). Relationships between selected properties of seeds, flours, and
starches from different chickpea cultivars. International Journal of Food Properties, 9(4), 597–
608. https://doi.org/10.1080/10942910600853774
8. Kawongolo, J. B. (2013). Optimization of Processing Technology for Commercial Drying of
Bananas ( Matooke ), 1 / 140.
9. Manzi, P., Aguzzi, A., & Pizzoferrato, L. (2001). Nutritional value of mushrooms widely
consumed in Italy. Food Chemistry, 73(3), 321–325. https://doi.org/10.1016/S0308-
8146(00)00304-6
10. Manzi, P., Gambelli, L., Marconi, S., Vivanti, V., & Pizzoferrato, L. (1999). Nutrients in edible
mushrooms: An inter-species comparative study. Food Chemistry, 65(4), 477–482.
https://doi.org/10.1016/S0308-8146(98)00212-X
11. Muranga, F. I., Mutambuka, M., Nabugoomu, F., & Lindhauer, M. G. (2010). Optimisation of
raw tooke flour , vital gluten and water absorption in tooke / wheat composite bread : Effect of
raw tooke flour and vital gluten on wheat flour physicochemical and dough rheological
properties ( Part I ). Journal of Food Science, 4(May), 223–230.
12. Nyombi, K., Van Asten, P. J. A., Leffelaar, P. A., Corbeels, M., Kaizzi, C. K., & Giller, K. E.
(2009). Allometric growth relationships of East Africa highland bananas (Musa AAA-EAHB)
cv. Kisansa and Mbwazirume. Annals of Applied Biology, 155(3), 403–418.
https://doi.org/10.1111/j.1744-7348.2009.00353.x
13. Pandey, D., & Kulshrestha, K. (2003). Carotene Rich Food Supplements Based On Carrot
Powder.Asian Journal Of Dairy and Food Research, 22(3).

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14. Ravi, U., Lakshmi, M., & Ranjani, M. (2010). Nutritional , sensory and physical analysis of
pumpkin flour incorporated into weaning mix Nutritional , Sensory and Physical Analysis of
Pumpkin Flour Incorporated into Weaning Mix. Malaysian Journal of Nutrition, 16(3), 379–
387.
15. Shimelis, E. A., Meaza, M., & Rakshit, S. K. (2006). Physico-chemical Properties, Pasting
Behavior and Functional Characteristics of Flours and Starches from Improved Bean (Phaseolus
vulgaris L.) Varieties Grown in East Africa. The CIGR Ejournal, III, 1–19.
16. Tibagonnzeka, J., Wambete, J., Muyimda, A., & Muyonga, J. (2014). Acceptability and
nutritional contribution of grain amaranth recipes in Uganda. African Journal of Food,
Agriculture, Nutrition and Development, 14(3), 8979–8997.

57
CHAPTER FOUR: RESEARCH MANUSCRIPT TWO

Optimization of extrusion conditions and determination of the effects of incorporation of cooked

vegetable-chicken mixture on sensory acceptability, nutritional composition and physico-
chemical properties of the instant soup powder.

Abstract

The effect of feed moisture content and barrel temperature on product responses which include; vitamin
A retention, vitamin C, viscosity, total fats, were studied using response surface methodology. The
mixture of 50:50 banana-Amaranth was extruded at different moisture content (12–20%) and barrel
temperature (120-180°C).The results were analyzed for easy reconstitution using boiled water and a
flour rate of 8% (80g of flour into 1L of water) from which some samples which could not reconstitute
were eliminated. To improve acceptability, the samples were blended with chicken-vegetables mixture
that was cooked conventionally and dried to get flour. There is therefore, a need to develop nutritious
and instant foods. However, little information exists on the production of instant soup from banana
based composite flour and the effect of extrusion variables on banana-amaranths based flour is not
known. This study was therefore aimed at producing nutritious instant banana-amaranth based soups
and to determine the effect of extrusion variables (feed moisture content and barrel temperature) on the
nutritional quality and overall acceptability of the products. This study achieved the objective of
contributing to the nutrition status of children under-five years of age.

4.1. Introduction

The global lifestyle of people, characterized by limited free time and increased working hours, has led
to an increased demand of RTE products(Filli, Nkama, & Jideani, 2013). Extrusion cooking technology
has been employed in developing RTE products from raw materials ranging from cereal flour, tubers
and legumes. Examples of extruded products include breakfast cereals, snacks, porridges, soups, flakes
and quickcooking pasta(James & Nwabueze, 2013).Like other thermal processing methods, extrusion
has effects on nutritional and physicochemical properties(Nayak et al., 2011).The destruction of pro-
vitamin Awas reported to be low compared to conventional traditional methods due to HTST
technology explored during extrusion(Singh et al., 2007). The destruction of pro-vitamin A depends
onthe extrusion condition; moisture content, screw speed, feed rate and temperature(Pathania et al.,
2013). Extrusion cooking is a known technology used to produce instant foods of good sensory quality

58
for children(Muoki, 2013). Extrusion offers hope for improving nutrition in less-developed
nations(Camire, 2001). The process enhances protein digestibility of foods through protein
denaturation, unfolding of polypeptide bonds and reduction in anti-nutritional factors (Björck & Asp,
1983). It also improves mineral bioavailability(Alonso et al., 2001) and results in higher vitamin A
retention. Previous research on extrusion cooking of instant soups has been explored to analyze the
effect of extrusion on vitamin A retention (Pathania et al., 2013; Pelembe et al., 2002).

4.2. Materials and methods

4.2.1. Materials

The materials presented in table 4.1were purchased from Capital shoppers‟ super market, garden city
branch, Kampala Uganda. The materials were parked and transported in paper bags and polythene
papers. The materials were first stored in a refrigerator to be prepared later in the afternoon.
Table 4.1: Formulations for preparation of chicken vegetable mix
Formulation

Component 1 2 3

Chicken 0.814kg 0.858kg 0.888kg

Water 2500 mls 2500 mls 2500 mls
Pumpkin 600g 600g 600g
Carrots 20g 20g 20g
Cooking time 2h 2h 2h
Onion 15g 15g 15g
Rosemary 0.5g of leaves 0.5g, leaves 0.5g, leaves
Thyme 1 sprig 1 sprig 1 sprig
Basil 0.5g of leaves 0.5g of leaves 0.5g of leaves
Dill pp 1 stalk 1 stalk 1 stalk
Bay leaves 5g 5g 5g
Parsley 5g 5g 5g
Gallic 7g 7g 7g
Chicken left after cooking 0% 10% 20%

59
 4.2.2. Methods

4.2.2.1. Method of preparation

The samples are weighed in the respective amounts using a Newal Kitchen Scale weighing scale
(Model No: NWL-7001, Turkish Origin) and put in an Aluminium source pan. The samples are boiled
using fire set using total gas for one hour. The pumpkin and carrot samples are then added to continue
the boiling for about 30 minutes. The boiled samples are left to simmer for 45 minutes. The samples are
then laid on an Aluminium foil and dried in an air Oven drier for about 48 hours. The dried samples are
then milled using a Pigeon mixer grinder (Classical lite, Stove Kraft Pvt. Ltd, India made). They are
then sieved using a 450 MIC sieve. In sample set 1, all the chicken pieces were removed while in
sample set 2 and 3, a significant amount of chicken was left to account for the 10% and 20% of the
total mass content respectively.
Table 4.2: Coding of flour vegetable-chicken mixes.
Chicken composition

Flours 0% 10% 20%

S1 50:50 X1 X2 X3
60:40 X11 X22 X33

S2 50:50 Y1 Y2 Y3
60:40 Y11 Y22 Y33

The ratios are in the form of F:C where F refers to extruded flour and C refers to chicken flour. Chicken
composition refers to the percentage of chicken meat dried with the vegetables. Samples S1 and S2 are
extruded flour samples at different extrusion conditions. Sample, S1 was extruded at an outlet
temperature of 120oC and moisture content of 10.17%. Sample, S2 was extruded at an output
temperature of 140oC and a moisture content of 14.3%. In all cases the screw speed, federate and
cutting rate were kept constant at 45Hz, 40Hz and 50Hz respectively. The produced samples shown in
table 4.2 were taken to the laboratory at SFTNB for nutritional, textural and physicochemical analysis.
4.2.2.2. The central composite design used to optimize levels of feed moisture content,
screw speed and the barrel temperature of the extruded flours

A central composite design (Table 4.3) were used to determine the effect of feed moisture content and
barrel temperature on the nutritional quality and consumer acceptability of Banana based flours. Each
60
combination of feed moisture content and barrel temperature were investigated first for reconstitution
and were then evaluated.

Table 4.3: Central Composite Design used to optimize feed moisture, screw speed and barrel
temperature for the production of instant banana-based soup flours

Run A: Barrel Temperature B: Feed Moisture

(oC) (%)
1 120 10.12
2 140 14.3
3 160 17.86
4 180 15.71
The other parameters of the extruder were kept constant include the feed screw speed, feeding rate,
cutting rate maintained at 45Hz, 40Hz and 50Hz respectively.
4.2.2.3. Reconstitution experiment to determine the most acceptable flours

Samples extruded at conditions in table 4.3 were prepared by boiling water and mixing with extruded
flour at a flour rate of 8g in 100mls of water (8%). The samples were thoroughly mixed and poured on
Aluminium foils for observation. Sample S1 and S2 were easy to reconstitute compared to samples S3
and S4. The pictorial effects have been illustrated as shown in figure 4.1 below
S1 S2 S3 S4

Figure 4.1: Reconstitution of extruded samples at different conditions

4.2.2.4. Effect of extrusion

Effect of extrusion on the dependent parameters were analyzed. Data were presented in form of models
and graphs while using the Desirability Function Approach, the best models were selected and the
effects of the parameters discussed.The dependent variables were expressed individually as a function
of the independent variables. The data was fitted to a second-order approximation model shown as
equation 4.1(Pathania et al., 2013):
61
Y  B0  i 1 Bi X i  i 1 Bii X i  i 1 Bij X i X j  
k k 2 k
(4.1)

Where Y is the response function, Xi is the feed moisture content, Xjis the barrel temperature, ε is the
random error, Bo the center point of the system, Bi, Bii, and Bij represent the coefficients of the linear,
quadratic and interactive effects of the independent variables respectively, and Xi, Xi2, and XiXj
represent the linear, quadratic and interactive effects, respectively of the independent variable

4.2.2.5. Preparation of composite flour soups for acceptability screening

Three different soups extruded at different conditions were prepared by mixing 50g of the flour in
800mls of water under constant stirring. Each composite flour was cooked until it boils up and then it
was allowed to simmer for five minutes. This was ready for serving to a panelist of about 30 people.

4.2.2.6. Sensory acceptability of the composite flour soups.

The soup mix was evaluated for its color, appearance, aroma and texture on a 9-point hedonic scale by
a trained panel of 30 people. The 9-point hedonic scale grading was as follows: “1" Dislike extremely,
"2" Dislike very much, "3" Dislike moderately, "4" Dislike slightly, "5" Neither like nor dislike, "6"
Like slightly, "7" Like moderately, "8" Like very much and "9" Like extremely. The samples were
coded and then presented to the panel for evaluation (Rodrigues et al., 2012).;

4.2.2.7. Nutritional and physicochemical properties

In order to determine the change in nutritional physicochemical and textural changes in the soup. The
generated samples were analyzed using standard methods described in section 3.6. Viscosity
measurements were carried out using the Brookfield viscometer.

62
 4.3. Results

Table 4.4: Consumer acceptability scores of soups from different flour formulations
Formulation Overall
code* Appearance Color Texture Aroma Mouth feel Taste acceptability
X1 6.53±1.94a 6.77±2.18a 6.53±1.41a 6.33±1.94a 6.80±1.65a 6.60±1.71a 7.20±1.58a
X11 6.53±1.83a 6.70±1.76a 6.57±1.45a 6.70±1.62a 6.43±1.63a 6.00±1.93a 6.97±1.77a
X2 6.93±1.11 a 6.67±1.49a 6.07±1.41a 6.43±1.48a 6.57±1.79a 6.93±1.57a 6.80±1.73a
X22 6.53±1.63a 6.13±1.55a 6.47±1.36a 7.13±1.81a 7.47±1.25a 7.17±1.42a 7.13±1.78a
X3 7.30±1.47a 7.00±1.44a 6.87±1.53a 6.73±1.17a 7.00±1.58a 7.43±1.19a 7.93±1.11b
X33 6.87±1.17a 6.30±1.24a 6.87±0.86a 7.13±1.41a 7.23±1.28a 7.40±1.16a 6.77±1.22a
Y1 6.67±1.27a 6.67±1.42a 6.63±1.45a 6.93±1.26a 7.07±1.60a 7.07±1.41a 7.30±1.02ab
Y11 7.17±1.37a 6.70±1.34a 6.70±1.29a 6.83±1.42a 7.07±1.44a 6.87±1.28a 7.20±1.67ab
Y2 7.00±1.62a 7.17±1.53a 6.63±1.47a 6.97±1.27a 6.43±1.33a 6.80±1.61a 6.77±1.10a
Y22 7.40±1.04a 6.37±1.19a 6.20±1.54a 6.60±1.50a 6.77±1.63a 7.17±1.21a 7.33±1.21a
Y3 6.50±1.98a 6.83±1.21a 6.37±1.45a 6.53±1.50a 6.77±1.57a 7.07±1.31a 7.07±1.41a
Y33 6.70±1.84a 6.73±1.36a 6.37±1.73a 6.00±1.26a 6.37±1.19a 6.90±1.95a 6.97±1.25a
Values represent mean ± standard deviation (n=30). Means in the same column with different
superscripts are significantly different (p < 0.05). *Formulation codes represent samples in the
respective formulations indicated in table 4.2
There is no significant difference between the sample formulations and appearance. Sample S10, has
the largest appearance of 7.4 while S0 has the least appearance of 6.13. There is no significant
difference between the sample appearance (p=0.05).The results for sensory analysis of soups made
from different flour mixtures of banana, grain amaranths and any of mushrooms, tomatoes, carrots,
pumpkins are presented in Table 4-6. For each sample, banana and amaranth flours were used fixed at a
proportion of 50:50. The formulation composition are as represented in table 4.2. The Control sample
S0 had the lowest scores on all the attributes. Sample S4 had the best scores on aroma and mouth feel
of 7.13 and 7.47 respectively which represents like moderately on the hedonic scale. The sample
formulation S5 had the best scores on taste and Overall acceptability of 7.43 and 7.93 which represents
like very much on the hedonic scale, thus the most preferred sample. The sample formulation S10 had
the best scores for appearance (7.40) while the sample formulation S9 had the best scores for color
(7.00) which represent like moderately on the hedonic scale. The test attributes all had the same

63
superscripts for all the formulations which depicts that all the attributes are not significantly different
from each other (p≤0.05). However, for appearance, color, texture, aroma, mouth feel, taste and overall
acceptability there is no statistically significant difference between the formulations at p ≤ 0.05.

4.3.1. Overall acceptability of the formulation

9
7.93
8 7.2 6.97 7.3 7.2 7.33
7.13 7.07 6.97
6.73 6.8 6.77 6.77
Overall acceptability

7
6
5
4
3
2
1
0
Sample codes
S0 S1 S2 S3 S4 S5 S6 S7 S8 S9 S10 S11 S12

Figure 4.2: Overall acceptability of the sample formulations

The overall acceptability of the samples was plotted against the sample codes as shown in figure
4.2above. The formulation code S5 has the most overall acceptability of 7.93 which represents like
very much on the hedonic scale. There are no significant difference between the attributes and Overall
acceptability of the samples. This means that the formulations are appealing to the humans for
consumption as a palatable food. The most preferred sample according to table 4.4 has a flour
composition of 40%, 40% and 20% banana, amaranth grains and pumpkins respectively. The next
preferred sample S10 has a composition of 45%, 45% and 10% respectively for banana, amaranth and
tomatoes while the next preferred S2 has a formulation of 42.5%, 42.5% and 15% respectively for
banana, amaranth and pumpkins. It can be concluded that formulations containing mushrooms are the
least acceptable followed by tomatoes, carrots and pumpkins respectively. Referring to the analysis of
variance table in the appendix section B, there is no statistical significance between groups of overall
acceptability and within the groups at p≤0.05. This shows that all the formulations can be considered
acceptable in front of the customer.

64
Table 4.5: Proximate composition of extruded samples
Sample Vit C Vit A RAE
code* MC (%) Fiber (%) Fat (%) Ash (%) (mg/100g) mg/100g
X1 9.32±0.01de 5.11±0.16bc 13.61±0.43cde 3.93±0.00a 9.26±0.96ab 0.74±0.02ab
Y1 8.53±0.03a 5.22±0.06c 12.72±0.82bc 4.14±0.03abcd 11.45±0.87cde 1.18±0.03e
X11 8.88±0.01bc 5.34±0.28cd 12.41±0.51bc 4.26±0.64abcd 10.71±0.85bcd 0.84±0.03abc
Y11 8.82±0.03ab 4.08±0.08a 13.01±0.82bcd 4.01±0.00ab 13.14±0.82e 0.97±0.01cd
X2 9.25±0.03d 3.67±0.21a 14.74±0.08ef 4.15±0.06abcd 9.79±1.16abc 0.76±0.02ab
Y2 9.20±0.04cd 3.68±0.01a 14.50±0.37de 4.36±0.18abcd 10.65±0.60bcd 0.76±0.00ab
X22 9.37±0.19de 5.76±0.26de 12.49±0.38bc 4.03±0.05abc 10.11±0.61bc 0.70±0.01a
Y22 9.47±0.10de 7.59±0.07f 12.25±0.96bc 4.19±0.26abcd 7.83±0.96a 0.98±0.25cd
X3 10.52±0.07g 4.61±0.12b 11.89±0.72b 4.75±0.11d 21.98±0.73g 1.08±0.04de
Y3 9.62±0.26ef 5.81±0.07de 16.38±0.45f 4.64±0.03cd 12.13±1.37de 1.12±0.07de
X33 10.21±0.16g 5.09±0.02bc 9.18±0.17a 4.57±0.14bcd 18.17±1.49f 1.08±0.27de
Y33 9.83±0.04f 6.02±0.32e 9.12±0.23a 4.42±0.00abcd 12.88±1.21e 0.87±0.01bc
Values represent mean ± standard deviation (n=3). Means in the same column with different
superscripts are significantly different (p<0.05). *Sample codes Xi (50:50) and Xii (60:40) represent
Sample 1 (S1)for Extruded flour: Vegetable Flour respectively while Yj (50:50) and Yjj (60:40)
represent Sample 2 (S2) for Extruded flour: Vegetable Flour respectively.

Generally, there are large variations in the results for the final developed product compared to those
that were obtained in objective one. A comprehensive comparison was desired to have a broader
picture of how this became an important variation for consideration.

65
 18 11

Moisture Content
Fat Content 16
10
14
12 9
10
8 8
0% 10% 20% 0% 10% 20%

Chicken Composition Chicken Composition

S1 (50:50) S2 (50:50) S1 (50:50) S2 (50:50)

S1 (60:40) S2 (60:40) S1 (60:40) S2 (60:40)

8
Fibre Content

4.8
7

Ash Content
4.6
6
4.4
5 4.2
4 4
3 3.8
0% 10% 20% 0% 10% 20%
Chicken composition Chicken composition

S1 (50:50) S2 (50:50) S1 (50:50) S2 (50:50)

S1 (60:40) S2 (60:40) S1 (60:40) S2 (60:40)

1.4 25
1.2
Vitamin A

Vitamin C

20
1
15
0.8
0.6 10
0.4 5
0% 10% 20% 0% 10% 20%
Chicken composition Chicken composition

S1 (50:50) S2 (50:50) S1 (50:50) S2 (50:50)

S1 (60:40) S2 (60:40) S1 (60:40) S2 (60:40)

Figure 4.3: Variations in nutrient composition; moisture content, fiber content, fat content, ash
content, Vitamin A and Vitamin C in the formulations
The moisture content varies between 8.53±0.03 and 10.52±0.07%. The graphical representation
explains the trend in the variation of moisture content with the samples (Figure 4.3). Moisture content
increases with composition of chicken in the samples. The addition of chicken thus leads to the storage
of more water molecules in the samples. The fiber content varies between 3.67±0.21 and 6.02±0.32%.
The graphical representation explains the trend in the variation of fiber content within the samples.
66
Fiber content improves with composition of chicken in the samples and is greater compared to
unextruded samples. The fat content varies between 9.12±0.23 and 16.38±0.45%. The graphical
representation explains the trend in the variation of fat content within the samples. Fat content increases
with composition of chicken in the samples. The addition of chicken thus leads to storage of more fat
molecules in the samples. There is a general increase in the fat content of the samples compared to
unextruded flour samples.The ash content varies between 3.93±0.00 and 10.52±0.07%. The graphical
representation explains the trend in the variation of ash content within the samples. Ash content
increases slightly with composition of chicken in the samples. The addition of chicken thus improves
on the quantity of ashes in the samples. The vitamin A content of the samples varies between 0.70±0.03
and 1.18±0.03 RAE mg/100g. The graphical representation explains the trend in the variation of
Vitamin A within the samples. Vitamin A increases with increases with composition of chicken in the
samples. There is a general improvement in Vitamin A related to the vitamin A that was obtained with
unextruded samples. The vitamin C content of the samples varies between 7.83±0.96 and 21.13±1.37
mg/100g. The graphical representation explains the trend in the variation of Vitamin C with the
samples. Vitamin C increases with increases with composition of chicken in the samples. There is a
general increase in Vitamin C related to the vitamin C that was obtained with unextruded samples.

4.4. Optimization of responses

Predictive models describing variability of dependent variables in the soups.

Responses Model Lack of

fit
2
Overall 7.12  0.02 X1  0.05 X 3  0.06 X 4  0.10 X1 X 3  0.11X1 X 4 R = 0.89, 0.1785
acceptability p=0.0001
2
Fat content 12.72  0.33 X1  0.2207 X 3  1.26 X 4  0.40 X1 X 3  0.23 X1 X 4 R = 0.90, 6.85
p=0.0001
Ash content 4.28  0.20 X 3  0.04 X 4  0.01X1 X 3  0.05 X 1 X 4 R2 = 0.96, 0.0414
p=0.0001
Fibre 5.08  0.15 X1  0.14 X 3  0.39 X 4  0.43 X1 X 3  0.07 X1 X 4 R2 = 0.86, 2.20
p=0.0001
Vitamin C 12.34  X1  0.12 X 3  0.20 X 4  2.15 X1 X 3  0.14 X1 X 4 R2 = 0.96, 9.93
p=0.0001
Vitamin A 0.92  0.05 X1  0.01X 3  0.02 X 4  0.1X1 X 3  0.03 X1 X 4 R2 = 0.84, 0.0722
p=0.0001
X1,=barrel temperature X2,=Moisture content, X3=Chicken vegetables mixand X4= banana-amaranth
flour

67
Barrel temperature (X1) and banana-amaranthflour (X4) had a negative linear effect on acceptability
whereas chicken composition (X3) had a positive linear effect on Overall acceptability of the soup. The
interactive effects were positive. The increase in temperature caused a significant reduction in
acceptability of the soup. Increase in chicken composition produces the soup by increasing the flavor in
the soup. Barrel temperature (X1) and chicken composition (X3) had a positive linear effect in the
composition of fats whereas banana-amaranth flour (X4) and interactions had a negative linear and
Quadratic effects on fat content. The increase in temperature and chicken composition had a significant
increase in the fat composition of the soup. Increase in chicken composition increases the soup‟s fat
content because of its high fat content. Barrel temperature (X1), Chicken composition (X3) and banana-
amaranth flour (X4) had significant linear and interactive effects on the ash content of the soups. The
increase in temperature and chicken composition caused a significant reduction in Ash content of the
soup. Barrel temperature (X1) and chicken composition (X3) had a negative linear and interactive
effects on fiber content of the soup whereas banana-amaranth flour (X4) had a positive linear effect on
Vitamin C composition of the soup. The increase in temperature caused a significant reduction in fiber
content of the soup. Barrel temperature (X1), chicken composition (X3) and banana-amaranth flour
(X4) had a negative linear effect and a positive interactive effectson Vitamin C composition of the
soup. The increase in temperature, chicken composition and banana flours reduces the Vitamin C
composition of the soup. Barrel temperature (X1) and chicken composition (X3) had a positive linear
and interactive effects on Vitamin a composition of the soup whereas banana-amaranth flour had a
negative linear and interactive effects on the soup. Increase in temperature increases the vitamin A
availability (Singh et al., 2007).

Table 4.6: Optimal solutions for nutritious and viscous soup formulations.
No. Temp MC CC BF VF OA FC Fat Ash Vit C Vit A Desirability

1 120.00 14.30 20 50 50 7.80 4.81 12.70 4.72 21.09 1.11 0.51 Selected

2 120.00 14.30 20 50 40 7.80 4.81 12.70 4.72 21.09 1.11 0.51

3 120.33 14.30 20 50 40 7.79 4.82 12.75 4.72 20.95 1.11 0.51

4 120.98 14.30 20 50 40 7.77 4.85 12.84 4.71 20.69 1.11 0.50

5 120.00 14.02 20 50 40 7.80 4.81 12.70 4.72 21.09 1.11 0.50

MC=moisture content, CC= Chicken level in the vegetable; BF=Banana-amaranth flour; VF=Vegetable
flour, OA=Overall acceptability, FC= Fiber content.
68
Figure 4.4: Response surface plots showing effect of varying different levels of moisture and temperature and chicken
composition on Vitamin A, Vitamin C, Ash content, Fat, Fiber content and Overall acceptability.

69
4.5. Conclusion and recommendations

The results indicated that there is a significant change (p=0.05) in texture as well as nutritional
composition and physico-chemical properties. The optimum extrusion conditions were temperature;
moisture combinations of 120oC and 14.3% respectively as shown in table 4.6.The banana flour was at
50% and a vegetable with chicken composition of 20% at 50%. The Overall acceptability, fiber
content, fat content, Ash content, Vitamin C and Vitamin A composition of the optimum flour was at
7.80, 4.81%, 12.70%, 4.72%, 21.09 g/100g and 1.11 mg/100g respectively.

4.6. References

1. Alonso, R., Rubio, L. A., Muzquiz, M., & Marzo, F. (2001). The effect of extrusion cooking on
mineral bioavailability in pea and kidney bean seed meals. Animal Feed Science and
Technology, 94(1–2), 1–13. https://doi.org/10.1016/S0377-8401(01)00302-9
2. Björck, I., & Asp, N. G. (1983). The effects of extrusion cooking on nutritional value - A
literature review. Journal of Food Engineering. https://doi.org/10.1016/0260-8774(83)90016-X
3. Camire, M. E. (2001). Extrusion and nutritional quality. In Extrusion Cooking (pp. 108–129).
https://doi.org/10.1533/9781855736313.1.108
4. Filli, K. B., Nkama, I., & Jideani, V. A. (2013). The Effect of Extrusion Conditions on the
Physical and Functional Properties of Millet – Bambara Groundnut Based Fura. American
Journal of Food Science and Technology, 1(4), 87–101. https://doi.org/10.12691/ajfst
5. James, S., & Nwabueze, T. U. (2013). Quality Evaluation Of Extruded Full Fat Blend Of
African Breadfruit-Soybean-Corn Snack. International Journal Of Scientific & Technology
Research Volume, 2(9).
6. Muoki, P. N. (2013). Nutritional, rheological and sensory properties of extruded cassava-soy
complementary porridges.
7. Nayak, B., Berrios, J. D. J., Powers, J. R., & Tang, J. (2011). Effect of Extrusion on the
Antioxidant Capacity and Color Attributes of Expanded Extrudates Prepared from Purple Potato
and Yellow Pea Flour Mixes. Journal of Food Science, 76(6). https://doi.org/10.1111/j.1750-
3841.2011.02279.x
8. Pathania, S., Singh, B., Sharma, S., & Sharma, V. (2013). Optimization of extrusion processing
conditions for preparation of an instant grain base for use in weaning foods. International
Journal of Engineering Research and Applications, 3(3), 1040–1049.
9. Pelembe, L. A. M., Erasmus, C., & Taylor, J. R. N. (2002). Development of a protein-rich
composite sorghum - Cowpea instant porridge by extrusion cooking process. LWT - Food
Science and Technology, 35(2), 120–127. https://doi.org/10.1006/fstl.2001.0812
10. Rodrigues, F. T., Fanaro, G. B., Duarte, R. C., Koike, A. C., & Villavicencio, A. L. C. H.
(2012). A sensory evaluation of irradiated cookies made from flaxseed meal. Radiation Physics
and Chemistry, 81(8), 1157–1159. https://doi.org/10.1016/j.radphyschem
11. Singh, S., Gamlath, S., & Wakeling, L. (2007). Nutritional aspects of food extrusion: A review.
International Journal of Food Science and Technology, 42(8), 916–929.
https://doi.org/10.1111/j.1365-2621.2006.01309.x

70
CHAPTER FIVE: RESEARCH MANUSCRIPT THREE

Estimating effect of incorporation of cooked chicken-vegetable mix on shelf-stability of the

instant soup flours using accelerated shelf-life testing.

Abstract

Most foods deteriorate with storage time thus the need to determine the time for which the flours are
going to remain fit for human consumption. The effect of incorporating chicken has been analyzed. In
order to reduce on the time spent in carrying out the experiment, ASLT was opted. This was done by
elevating the storage temperature to 37oC and then project the effect to room temperature.Twelve (12)
formulations extruded at two different temperature (120oC and 140oC): Moisture content combinations
mixed with carrot and pumpkin soup prepared with different ingredients and chicken wings at different
levels (0%, 10% and 20%) were subjected to same storage conditions. Microbial analysis of the flours
indicated that all samples contained recommendable counts for yeasts and molds, total coliforms and
total plate count, and were therefore considered good for human consumption for the storage period.

Keywords: Shelf life, microbial load, yeasts and molds, total plate count, total coliforms

5.1. Introduction

It is common for all processed foods to deteriorate during storage from their first date of production.
Determining the time for which the developed product shall remain acceptable for consumption without
causing harm to the consumer isvery important in new development. Before setting theexperiment for
shelf life testing, it is required to identify the indices of quality loss of a particularproduct. It is
important to study the deteriorative factors of the food product in order to predict shelf life(Villota et
al., 1992).Shelf life determination of a product normally involvesstoring the product under pre-selected
conditions for a period of time longer than the expectedshelf life and checking the product at regular
intervals to see when it begins to spoil(Labuza et al., 2002). This method requires aconsiderable
amount of experimentation making theprocedure costly and time consuming. Accelerated Shelf Life
Testing (ASLT) can be used toshorten the time of experimentation. With ASLT, food products are
stored at elevatedtemperatures to accelerate the deterioration process(Shalmany, 2012). Data
fromelevated temperatures are converted to regular storage conditions using Arrhenius or
Linear equations(Waghray,et al., 2017). The estimation of shelf lifeusing ASLT requires identification
of the major spoilage agent or reactions and selection of appropriate indices of spoilage. Therefore the

71
objective of this study was todetermine the effect of incorporating different levels of chicken to the
shelf-life of instantbanana-amaranths soups mixed with cooked vegetables.

5.2. Materials and methods

Accelerated shelf life testing was used to determine the shelf life of different formulatedsoups. Portions
(10g) of each sample were packed and sealed in transparent polyethylene bagsand stored at an elevated
temperature of 37oC.Sampling for analysis was doneat 0, 14, and 28 days(Waghray et al.,
2017).Microbial load was used as a basis to estimate the shelf life of the soup samples. Total coliforms,
yeasts and molds and total plate counts were determined and used as indices of quality loss during
storage (AOAC, 1995: method 965.33). The dataobtained from elevated temperatures was used to
estimate the shelf life at room temperature which are expected to vary between 23 °C and 27 °C
usingboth Arrhenius and Linear models as suggested by Lee & Krochta, (2002).

5.2.1. Yeasts and molds

Yeasts and molds were enumerated using the surface spread technique in ISO 21527 – 2:2009. About
15.8 g of Dichloran Rose Bengal Chloramphenicol (DRBC) agar (M588-500G, Hi-Media Laboratories
Pvt. Ltd.) was weighed and mixed with distilled water (500 ml). The mixture was sterilized by
autoclaving at 121oC for 15 minutes and rapidly cooled in a water bath to about 47oC. Molten agar (20
ml) was aseptically poured into the petri dish and allowed to set. The petri dishes were inverted to
avoid the dripping back of condensed water onto the solidified agar. Inocula (0.1 ml) from the serial
dilutions (10-1 to 10-7) of the soup flour samples were aseptically transferred onto the center of the
solidified agar and evenly spread over the surface of the agar using a sterile wire loop. This set up was
incubated at 37oC for 5 days. Plates with colonies between 30 and 300 were taken and counted using
the colony counter and results expressed as colony forming units per gram(cfu/g).

5.2.2. Total platecount

The total plate count was enumerated using the pour plate technique, ISO 4833:2013 method. Plate
count agar was prepared by dissolving 11.75 g of the agar powder (1056.00, CONDA Pronodisa
Laboratories Conda U.S.A) into distilled water (500 ml) and sterilized by autoclaving at 121°C for 15
minutes. It was then allowed to cool to about 47°C using a water bath (Grants instrument Ltd, Shepreth,
England). Serial dilutions (10-1 to 10-7) of the sample solutions were made. Inocula (1 ml) from each
selected dilution were aseptically transferred to petri dishes and about 20 ml of molten agar poured into

72
each petri dish containing the inoculum. This inoculum was carefully mixed with the agar by rotating
the petri dishes and then allowed to solidify. The dishes were then inverted after solidification of the
agar and incubated at 37°C for 24 hours. Plates with colonies ranging from 30 to 300 were considered
for counting using the colony counter (Stuart SC6, UK) and the results expressed as colony forming
units per gram (cfu/g).

5.2.3. Total coliforms count

The total coliforms were determined using the pour plate technique in ISO 4832:2006. Violet Red Bile
Lactose (VRBL) agar (M581-500G. Hi-Media Laboratories Pvt. Ltd.) was prepared by weighing 20.75
g of media powder into 500ml of distilled water. The mixture was heated on a Bunsen burner flame
until boiling and allowed to boil for 2 minutes then rapidly cooled to 47°C using a water bath. Serial
dilutions (10-1 to 10-7) of the soup flour samples was made and 1 ml of inoculum from each dilution
transferred aseptically to petri dishes. Molten agar (20 ml) was poured into each of the petri dishes
containing inocula and carefully mixed by rotating the petri dishes. After solidification, agar (5 ml) was
poured over the surface of the previously solidified mixture and left to solidify again to create an
anaerobic atmosphere. The dishes were then inverted and incubated at 37°C for 24 hours. After 24
hours, the purplish colonies formed which were considered to be typical coliform colonies. Petri dishes
with counts ranging from 30 to 300 were counted using the colony counter and results expressed in
colony forming units per gram (cfu/g).
5.2.4. Calculations

The microbial counts, represented as colony forming units per gram (cfu/g), were calculated using
equation 5.1 below

C X (5.1)
V n1  n2 (0.1)d

Where: C =microbial counts, ∑X = sum of all counted colonies, n1 = number of petri dishes at which
the first counting was done, n2 = number of petri dishes at which the second counting was done, d =
dilution factor at which the first counting was done.

73
5.3. Results and discussion

TC (log 10 Cfu/g)
10.00 8.00
TPC (log 10 Cfu/g)

8.00 6.00

6.00 4.00

4.00 2.00

2.00 0.00
0 14 28 0 14 28

Storage time (days) Storage time (days)

Y1 Y2 Y3 Y11
Y1 Y2 Y3 Y11
Y22 Y33 X1 X2 Y22 Y33 X1 X2
X3 X11 X22 X33 X3 X11 X22 X33

10.00
Y&M (log 10 Cfu/g)

8.00
6.00
4.00
2.00
0.00
0 14 28
Storage time (days)
Y22 Y33 X3 X33

Figure 5.1: Microbial changes in the soup samples during storage

74
5.3.1. Microbial load in the flours

5.3.1.1. Total plate count

There were significant differences in the total plate count of the flour samples during the high
temperature storage (P<0.05) as illustrated in figure 5.1. Generally, the total plate count in all the soup
flours decreased from day 0 to day 28. The reduction could have been contributed by the packaging
method and the high temperature storage. The differences in the microbial load could be due to the
different components in the soup. Microbial growth and metabolism is a major cause of flour spoilage
which produce amines, biogenic amines such as putrescine, histamine and cadaverine, organic acids,
sulphides, alcohols, aldehydes and ketones with unpleasant and unacceptable off-flavors (Dalgaardet
al., 2006; Emborget al., 2005; Gram and Dalgaard, 2002). The observed total plate counts were all
within acceptable limits; since 106cfu/g is the maximum permissible level for aerobic plate counts in
meat products (UNBS, 2015; ICMFS 2002). Therefore, all the soup flours analyzed over the period of
four weeks of storage at high temperature can be considered acceptable for human consumption

5.3.1.2. Total coliform

There were significant differences in the total coliforms in the storage of the flour samples during high
temperature storage for the entire period of 6 weeks storage as shown in figure 5.1. The population of
total coliforms in the soup flour samples were less than 6 log cfu/g at both day 0 and day 28. The total
coliforms decreased throughout the high temperature storage. The presence of total coliforms is an
indication of contamination by humans, contaminated water used during processing or ingredients used
during processing. Some of the spices used in the flours could also contain some potential contaminants
(Gungor & Gokoglu, 2010). However, spices such as garlic, and ginger used contain antimicrobial
agents such as allicin and gingerol respectively and therefore form natural preservatives in the food.

5.3.1.3. Yeasts and Molds

There were no yeasts and molds detected in most of the flours. Yeasts and molds in most of the flours
were in acceptable limits.

The Arrhenius model has been used very often to predict the storage time of most foods at different
temperatures (Dube, 2015). This was used to predict the storage time at any time.

5.4. Conclusion

75
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APPENDIX
A. ANOVA FOR SENSORY EVALUATION

Sum of Squares df Mean Square F Sig.

Between Groups 13.400 2 6.700 2.399 .097
Appearance Within Groups 243.000 87 2.793
Total 256.400 89
Between Groups 3.622 2 1.811 1.150 .321
Color Within Groups 137.000 87 1.575
Total 140.622 89
Between Groups .556 2 .278 .112 .895
Texture Within Groups 216.733 87 2.491
Total 217.289 89
Between Groups 6.489 2 3.244 1.598 .208
Aroma Within Groups 176.667 87 2.031
Total 183.156 89
Between Groups 3.200 2 1.600 .734 .483
Mouth feel Within Groups 189.700 87 2.180
Total 192.900 89
Between Groups 1.089 2 .544 .234 .792
Taste Within Groups 202.733 87 2.330
Total 203.822 89
Between Groups 2.156 2 1.078 .644 .527
Overall
Within Groups 145.500 87 1.672
acceptability
Total 147.656 89

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 B. ONE WAY ANOVA FOR PROXIMATE COMPOSITION

Sum of Squares df Mean Square F Sig.

Between Groups 13.998 12 1.167 9.567 .000
MC Within Groups 3.170 26 .122
Total 17.168 38
Between Groups 7.979 12 .665 11.269 .000
Fat Within Groups 1.534 26 .059
Total 9.513 38
Between Groups 11.721 12 .977 26.234 .000
Fiber content Within Groups .968 26 .037
Total 12.689 38

C. Production of dessert and Cooking Bananas in Uganda (Source FAOSTAT, 2015)

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D: 3D PLOTS AND 2D CONTOURS FOR RSM
4.1.8 Gross Energy Content

4.1.9 Ash Content

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4.1.10 Vitamin C

4.1.10 Vitamin A

4.1.10 Carbohydrates

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4.1.10 Final Viscosity (Cp)

4.1.10 Peak time

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4.1.10 Starch Content

4.1.10 Iron Content

4.1.10 Potassium

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92
Pasting profile for the soup flours

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