Micronutrients in Moringa oleifera and their Potential in Food

Fortification

by

W
IE
Yee Kei Kiki Chan
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A thesis submitted in conformity with the requirements

for the degree of Master of Applied Science
Department of Chemical Engineering and Applied Chemistry
University of Toronto

c Copyright 2018 by Yee Kei Kiki Chan





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 Abstract
Micronutrients in Moringa oleifera and their Potential in Food Fortification

Yee Kei Kiki Chan

Master of Applied Science
Chemical Engineering and Applied Chemistry
University of Toronto
2018

Moringa oleifera is frequently endorsed for its high micronutrient content relative to other

vegetables, but reported data on Moringa’s nutritional value are inconsistent with common

nutritional claims. A comparative analysis on the macronutrient and micronutrient content

of Moringa leaves and pods was conducted. Moringa contains multiple nutrients but its iron

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content is similar to that in spinach and its vitamin A content is lower than carrots on a dry

basis. Nevertheless, Moringa’s abundance in micronutrient deficient regions makes it applica-

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ble as a natural fortificant. Bouillon cubes were identified as a suitable food vehicle for the

incorporation of Moringa. Bench-scale cold extrusion processing was conducted to explore the
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feasibility of creating Moringa-fortified bouillon cubes. Nutritional characterization confirmed

that supplemental fortificants would be required to enhance the nutritional value and stability

of Moringa-fortified bouillon cubes. Flavours of Moringa may either be enhanced or suppressed

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depending on the application.

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To gung gung
IE (1924-2016)
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 Acknowledgements
I would like to express my gratitude towards my co-advisors, Professor Levente Diosady and
Professor Yu-Ling Cheng. Thank you for mentoring me, for teaching me effective ways to frame
and approach research problems (I am still working on this), and for the incredible opportuni-
ties you’ve given me at UofT and beyond. I would also like to thank my committee members,
Professor M.G. Venkatesh Mannar and Professor Radhakrishnan Mahadevan, for their helpful
insights on this thesis topic.

Heartfelt thanks to friends and family who eagerly listened to numerous recounts of small vic-
tories and encouraged me to grow with every challenge presented.

To Mom and Dad: Thank you for your limitless love and support. Thank you for teaching
me to be grateful and to count my blessings. Thank you for taking care of everything else so I

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could focus on whatever I chose to do.

To Nickie: Thank you for the inside jokes and for cheering me up always. Stay close, BB :)
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To Elisa, Kiruba, Segun, Juveria, Azadeh, Folake and Rahul: Thank you for celebrat-
ing with me when things went well and for urging me to keep going when things went wrong.
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I’m so lucky to get to work (or have worked) alongside you all at the Food Engineering lab.

To Jonathan: Thank you for showing me a glimpse of God’s love on earth. Thank you for
keeping me grounded, for being a great friend and companion. Marrying you is one of the best,
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if not the best, decision I’ve made in my life. I love you so much.

And lastly...

Everything good within me comes from God. Without Him, I am nothing and all
my works are empty.

Beloved, let us love one another, for love is from God, and whoever loves has been born of God
and knows God. Anyone who does not love does not know God, because God is love. In this the
love of God was made manifest among us, that God sent his only Son into the world, so that
we might live through him. In this is love, not that we have loved God but that he loved us and
sent his Son to be the propitiation for our sins. — 1 John 4:7-10

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Contents

Dedication iii

Acknowledgements iv

Contents v

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List of Tables vii

List of Figures

Acronyms
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1 Background 1
1.1 Global Food Insecurity and Micronutrient Deficiencies . . . . . . . . . . . . . . . 1
1.2 Nutritional Content and Requirement Standards . . . . . . . . . . . . . . . . . . 3
1.3 Food Fortification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
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1.3.1 Existing Fortification Interventions . . . . . . . . . . . . . . . . . . . . . . 4

1.3.2 Fortificants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
1.4 Moringa oleifera as a Natural Food Fortificant . . . . . . . . . . . . . . . . . . . 6
1.5 Scope of Thesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
1.6 Objectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

2 Moringa oleifera Nutritional Characterization 10

2.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
2.2 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
2.2.1 Materials and Sample Preparation . . . . . . . . . . . . . . . . . . . . . . 11
2.2.2 Proximate Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
2.2.3 Mineral Content . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
2.2.4 Vitamin Content and Protein Quality . . . . . . . . . . . . . . . . . . . . 12
2.3 Results and Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
2.3.1 Proximate Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
2.3.2 Mineral Content . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

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 2.3.3 Vitamin content . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
2.3.4 Protein Quality . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
2.4 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

3 Addressing Micronutrient Deficiencies using Moringa oleifera 26

3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
3.2 Existing Fortification Interventions using Moringa oleifera . . . . . . . . . . . . . 26
3.2.1 Yogurt . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
3.2.2 Bread . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
3.2.3 Cookies and Extruded Snacks . . . . . . . . . . . . . . . . . . . . . . . . . 27
3.2.4 Region-specific Dishes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
3.2.5 Raw Meat and Fruit Juices . . . . . . . . . . . . . . . . . . . . . . . . . . 29
3.3 Overcoming Technical Challenges . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
3.4 Food Vehicle Options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

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3.4.1 Consumption Coverage . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
3.4.2 Shelf-life . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
3.4.3 Ease of Adding Other Micronutrients . . . . . . . . . . . . . . . . . . . . . 33
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3.4.4 Public Health Concern . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
3.5 Proof of concept for Moringa-fortified bouillon cubes . . . . . . . . . . . . . . . . 33
3.5.1 Commercially Available Bouillon Cubes . . . . . . . . . . . . . . . . . . . 34
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3.5.2 Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
3.5.3 Experimental Design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
3.5.4 Processing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
3.5.5 Characterization Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
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3.5.6 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
3.6 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47

4 Future Work and Conclusions 48

4.1 Future Work . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
4.1.1 Taste, Aroma and Colouring Compounds . . . . . . . . . . . . . . . . . . 48
4.1.2 Encapsulation and Coating Processes . . . . . . . . . . . . . . . . . . . . 49
4.1.3 Alternative Binders and Excipients . . . . . . . . . . . . . . . . . . . . . . 49
4.1.4 Supplemental Fortificants . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
4.1.5 Manufacturing Process Selection and Optimization . . . . . . . . . . . . . 50
4.1.6 Consumption Patterns and Consumer Preferences . . . . . . . . . . . . . 50
4.2 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51

5 Appendix 53

Bibliography 68

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List of Tables

1.1 Common micronutrient deficiency symptoms and affected vulnerable population

groups . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
1.2 Nutritional content standards, RDA and NRV . . . . . . . . . . . . . . . . . . . . 3
1.3 Production yields and harvest periods for perennial and annual Moringa oleifera 7

2.1 Moringa samples for experimental analyses . . . . . . . . . . . . . . . . . . . . . 11

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2.2 Proximate compositions of Moringa leaves and pods. Rows marked dashes (-) in
the reference column were experimentally determined in this project. . . . . . . . 13
2.3 Moisture content, dry mass and protein content in Moringa pods skin and flesh . 13
2.4
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Dietary Reference Intakes for select minerals (19-50 years old) [1] . . . . . . . . . 15
2.5 Mineral content in Moringa leaves and pods. Rows marked with dashes (-) in
the reference column were measured in this project. . . . . . . . . . . . . . . . . . 15
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2.6 RDA for select vitamins (19-50 years old) [2] . . . . . . . . . . . . . . . . . . . . 23
2.7 Vitamin content in Moringa leaves and pods (literature values) . . . . . . . . . . 23
2.8 Protein quality and amino acid scores expressed as percentage of WHO Adult
requirement in Moringa leaves and pods . . . . . . . . . . . . . . . . . . . . . . . 24
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3.1 Evaluation of food vehicles for incorporating Moringa . . . . . . . . . . . . . . . 31

3.2 Comparison of NRV and RDA values for minerals, vitamins and protein . . . . . 35
3.3 Physical specifications for fortified bouillon cubes . . . . . . . . . . . . . . . . . . 36
3.4 In-barrel moisture content in screening experiments . . . . . . . . . . . . . . . . . 40
3.5 In-barrel moisture for mixture design formulations . . . . . . . . . . . . . . . . . 40
3.6 %NRV of iron, zinc, vitamin A, folate and protein per 3.3g serving . . . . . . . . 42
3.7 Mass, hardness, water activity and disintegration results for extruded cubes . . . 44
3.8 Hunter L*ab values for extruded cubes . . . . . . . . . . . . . . . . . . . . . . . . 46

5.1 Essential amino acids (mg/g protein) in Moringa leaves and pods (literature values) 54
5.2 Mineral content of extrudates per 3.3g serving or Moringa-fortified cubes . . . . 54

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List of Figures

2.1 Protein content in Moringa leaf and pod samples measured in this study . . . . . 14
2.2 Iron content comparisons in Moringa leaf samples. Samples marked with aster-
isks (*) were experimentally determined in this study. Dashed lines indicate the
RDA for individuals between 19 and 50 years old. . . . . . . . . . . . . . . . . . . 16
2.3 Mineral content comparisons in Moringa leaf samples (calcium, magnesium,

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potassium and sodium). Samples marked with asterisks (*) were experimen-
tally determined in this study. Dashed lines indicate the minimum RDA or AI
for individuals between 19 and 50 years old. . . . . . . . . . . . . . . . . . . . . . 17
2.4
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Mineral content comparisons in Moringa leaf samples (zinc, copper and man-
ganese). Samples marked with asterisks (*) were experimentally determined in
this study. Dashed lines indicate the minimum RDA for individuals between 19
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and 50 years old. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
2.5 Iron content comparisons in Moringa pod samples. Samples marked with aster-
isks (*) were experimentally determined in this study. Dashed lines indicate the
RDA for individuals between 19 and 50 years old. . . . . . . . . . . . . . . . . . . 19
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2.6 Mineral content comparisons in Moringa pod samples (calcium, magnesium,

potassium and sodium). Samples marked with asterisks (*) were experimen-
tally determined in this study. Dashed lines indicate the minimum RDA or AI
for individuals between 19 and 50 years old. . . . . . . . . . . . . . . . . . . . . . 20
2.7 Mineral content comparisons in Moringa pod samples (zinc, copper and man-
ganese). Samples marked with asterisks (*) were experimentally determined in
this study. Dashed lines indicate the minimum RDA for individuals between 19
and 50 years old. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
2.8 Percentage of total minerals in the skin of Moringa pods . . . . . . . . . . . . . . 22

3.1 Ingredient composition in a commercial bouillon cube . . . . . . . . . . . . . . . 34

3.2 Simple axial mixture experimental design for proof of concept . . . . . . . . . . . 37
3.3 Block flow diagram for forming Moringa-fortified bouillon cubes . . . . . . . . . . 38
3.4 (left) Cold extrusion setup. Die attached to single Archimedes screw attachment;
(right) Stainless steel extrusion die with 20*20mm square opening . . . . . . . . 39

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3.5 Iron content in mixture design formulations. Increasing bubble sizes correspond
to higher levels of iron; numbers correspond to mean iron content per cube
(mg/3.3g serving). Iron NRV = 14mg/d; benchmark was 15% NRV = 2.1mg . . 43
3.6 Hardness in mixture design formulations. Increasing bubble sizes correspond to
increasing hardness; numbers correspond to mean hardness in newtons. The
range for acceptable hardness is 5-50N. . . . . . . . . . . . . . . . . . . . . . . . . 45
3.7 Formation of cracks (top face) after drying suggesting uneven extrusion pressure.
Sample of cube made with 1:1 ratio of Moringa leaves and binder. . . . . . . . . 45

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Acronyms

AI Adequate Intake

AOAC Association of Official Analytical Chemists

ANOVA Analysis of variance

ASTM American Society for Testing and Materials

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DFE Dietary Folate Equivalent

DRI Dietary Reference Intakes

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FAO Food and Agriculture Organization of the United Nations
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FDA United States Food and Drug Administration

GC-MS gas chromatography-mass spectrometry

GDP Gross Domestic Product

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HSD Honest Significant Difference

ICP-AES Inductively coupled plasma - atomic emission spectroscopy

IoM Institute of Medicine

LMIC Low and middle-income country

MSG Monosodium glutamate

n.d. Not determined

NRV Nutrient Reference Value

PPP Purchasing Power Parity

RAE Retinol Activity Equivalent

RDA Recommended Dietary Allowance

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SDG Sustainable Development Goal

UL Tolerable Upper Intake Level

WHO World Health Organization

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Chapter 1

Background

This chapter presents an overview of global micronutrient deficiencies and approaches that
have been investigated or implemented to address this global challenge. The background pro-

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vided on Moringa oleifera and its potential in addressing micronutrient deficiencies leads to the
motivation and scope of this thesis.

1.1
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Global Food Insecurity and Micronutrient Deficiencies
Definitions of food security have evolved as the understanding of the relationship between
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food and well-being deepened. Up until the 1970s, food security referred to a nation’s ability
to balance its caloric supply and demand, which overlooked numerous intricacies of access,
distribution, and nutrition [3, 4]. Since 1974, the Food and Agriculture Organization of the
United Nations (FAO) has defined food security as “A situation that exists when all people, at
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all times, have physical, social and economic access to sufficient, safe and nutritious food that
meets their dietary needs and food preferences for an active and healthy life” [5]. The inclusivity
of this updated definition allows food insecurity to be examined at all stakeholder levels—from
individuals to communities to economies—and as a multifaceted global challenge. The explicit
reference for adequate nutrition points directly to the need to address micronutrient deficiencies,
commonly called “hidden hunger”, which affects both developing and developed nations, albeit
unequally [6]. The importance of addressing global micronutrient deficiencies is also highlighted
by the United Nations in Sustainable Development Goal (SDG) #2: End hunger, achieve food
security and improved nutrition and promote sustainable agriculture [7].
In this thesis, food security is viewed as a prerequisite for nutrition security. One of the main
determinants for nutrition security is adequate micronutrient intake. Although it is possible
that nutrition security could exist without food security (e.g. taking micronutrient supplements
without adequate caloric intake), such cases are rare in low and middle income countries; it is
more typical that households would need to be able to satisfy caloric requirements before being
able to consider the nutrition within the foods consumed. The relation between food security
and nutrition security continues to be debated in literature [8]. Nevertheless, differing opinions

1
Chapter 1. Background 2

in definitions do not negate the importance of addressing global micronutrient deficiencies.

Micronutrient deficiencies affect individuals, communities and economies alike, and are
linked to the perpetuation of the poverty cycle, where constant burdens of fatigue and disease
lead to the inability to work and earn wages for survival [9]. Although significant improvements
have been observed for caloric deficiencies since the year 2000, micronutrient deficiencies re-
main prominent and widespread [5]. A conservative estimate is that symptomatic micronutrient
deficiencies affects two billion people globally, as asymptomatic cases of mild deficiencies are
difficult to diagnose and quantify [3, 10, 11]. Public health indicators such as anemia, stunting,
and night blindness suggest that micronutrient deficiencies are most prevalent in African and
South Asian regions [5, 12].
Deficiencies in iron, vitamin A, iodine, folate, zinc, and vitamin B12 [13, 14] are most
prevalent. Symptoms, signs and complications of micronutrient deficiencies are most prominent
in pregnant women in whom reproductive health and fetus development is impeded, and in
children in whom growth retardation is observed. Table 1.1 summarizes the impacts of common

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micronutrient deficiencies and demonstrates the importance of adequate micronutrient intake
for health [14–16]. Economic impact estimates range from 2% Gross Domestic Product (GDP)
loss associated with iron deficiency [14] to 5% overall GDP loss associated with vitamin A,
iodine and iron deficiencies [17].
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The World Health Organization (WHO) promotes three main strategies for increasing the
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intake of micronutrients: diversification, supplementation and fortification [10]. Public health
organizations mandate and implement combinations of these strategies in parallel to form an
integrated approach for addressing micronutrient deficiencies. Those living in low and middle
income countries are most affected by food insecurity since poorer regions experience food
shortages, and are unable to balance micronutrient intakes through diet diversification [6, 18].
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Table 1.1: Common micronutrient deficiency symptoms and affected vulnerable population
groups
Micronutrient Essential for functioning of Symptoms Indicating Moderate to Severe Deficiency Most Affected Population Groups
Goiter Pregnant women
Iodine Thyroid hormones Mental retardation Newborns and infants
Pregnancy complications Vegetarians
Chronic fatigue Women of child-bearing ages
Iron Hemoglobin and myoglobin Heart failure Children
Pica Vegetarians
Blindness Newborns
Vitamin A Eyes and immune system Stunted growth Pregnant women
Frequent infections Children
Chronic fatigue Pregnant and lactating women
Folate Amino acid synthesis Neural tube defects Newborns and infants
Stunted growth People with alcoholic dependence
Frequent infections Gastrointestinal disease patients
Zinc Immune system and amino acid synthesis Stunted growth Vegetarians
Loss of appetite Pregnant and lactating women
Chronic fatigue Elderly
Vitamin B12 Amino acid synthesis Heart failure Pregnant and lactating women
Numbness in limbs Vegetarians
Chapter 1. Background 3

1.2 Nutritional Content and Requirement Standards

Nomenclature for the reference values for nutritional requirements and benchmarks differ de-
pending on the region and application. Two different types of reference values are used in
this thesis: Recommended Dietary Allowance (RDA) and Nutrient Reference Value (NRV).
In general, RDA is used by health professionals, such as dieticians, to assess the nutritional
requirements of an individual; whereas NRV is used to benchmark the level of micronutrients
in a food product (Table 1.2). Accordingly, Chapter 2 (Moringa oleifera Nutritional Charac-
terization) of this thesis will reference RDA values, and Chapter 3 (Addressing Micronutrient
Deficiencies using Moringa oleifera) of this thesis will reference NRV for benchmarking.

Table 1.2: Nutritional content standards, RDA and NRV

Recommended Dietary Allowance Nutrient Reference Value
(RDA) (NRV)
Originator Institute of Medicine Codex Alimentarius Commission

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Methodology Sufficient intake for 97-98% of the target population group Sufficient intake for 97.5% of the entire population
Value Format mg/d; values split into different population groups according to age and sex mg/d; single value for entire population
Application Health impact; nutritional requirements Benchmarking of fortification programs; nutrition labelling and claims

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1.3 Food Fortification
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Food fortification is the process of adding nutrients to food, which may be mandated or per-
formed on a voluntary basis. Mandatory food fortification programs are implemented in regions
to address micronutrient deficiencies of public health concern, whereas voluntary fortification
is often performed so that a health claim may be made for marketing purposes. Food fortifi-
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cation programs are proven to be effective for reducing the prevalence of several micronutrient
deficiencies relatively quickly, sustainably, and economically [19–21].
Industrial fortification of various staple foods and condiments has been successful in reme-
diating micronutrient deficiencies and have considerable benefits [11]. First, staple foods and
condiments are widely consumed by significant portions of the population, so high household
coverage can be achieved. Second, staple foods and condiments that are centrally produced by a
small number of large manufacturers allow the quality and level of fortification to be controlled
and monitored. Finally, minimal behavioural changes in dietary habits are required as long as
there are no changes in organoleptic properties of fortified foods is detectable to the consumer.
Significant challenges associated with food fortification are: determining the appropriate
fortification level, cost, stability of fortificants—particularly due to possible interactions among
multiple fortificants, and maintaining organoleptic properties. Considering the varying micronu-
trient requirements within a population and even within a single household, it is challenging
to determine the amount of fortificant for a food vehicle that is appropriate for all potential
consumers [20, 21]. If fortification levels are set too low, the fortified food will have little to
no observable effect on reducing the targeted micronutrient deficiencies. If fortification levels
Chapter 1. Background 4

are set too high, the excessive micronutrient intake may become toxic and detrimental to an
individual’s health. Although fortification is considered a cost-effective intervention strategy,
additional processing still inevitably increases the cost of a food product. This is especially
burdensome for the poorest of the poor, who are extremely price sensitive and are likely to buy
lower quality, inadequately fortified versions of the same food [10, 21]. In the development of
food vehicles fortified with multiple micronutrients, undesirable interactions leading to reduced
bioavailability and stability are common technical challenges. Similarly, maintaining the orig-
inal organoleptic properties are also challenging due to interactions between the food vehicle
and fortificant and is sometimes a major barrier for the commercialization of fortified food [10].
There are also challenges in quality control and cost containment for fortified foods. The
level of fortified foods is likely to be higher and more consistent when fortification is implemented
for food vehicles with a few key manufacturers as opposed to food with a great number of smaller
scale manufacturers [22]. The level of food fortification is also more consistent in a mandatory
environment where appropriate monitoring systems are in place [23, 24]. Additionally, large

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manufacturers are more likely able to reach economies of scale, and to contain additional costs
associated with fortification processes.
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1.3.1 Existing Fortification Interventions
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Salt Fortification

Salt was among the first food vehicles to be commercially fortified in the 1920s to reduce goiter
occurrences in Switzerland and the United States [25, 26]. Salt is considered an ideal food
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vehicle for fortification as it is universally consumed with minimal variations in consumption

levels amongst households of different socioeconomic statuses [27, 28]. Salt iodization is the
preferred method for controlling iodine deficiency disorders and there has been ongoing research
to fortify salt with other micronutrients [27].
Implementation trials of double fortified salt with iodine and iron has demonstrated promis-
ing results in reducing iron deficiency anemia in low and middle-income countries [29, 30]. To
produce double fortified salt, a premix of ferrous particles are agglomerated with the aid of
binders in cold extrusion, encapsulated, and blended with iodized salt [31, 32]. Encapsulation
provides both a colour-masking layer that covers the red-brown colour of the ferrous particles
and a physical isolation barrier for stability.
Other approaches to fortify salt with multiple micronutrients are also under development.
Attempts were made to add vitamin A to produce triple fortified salt but were not commercial-
ized due to technical barriers in stability [33] and high energy requirement of the production
processes [34]. A variety of projects are ongoing to fortify salt with combinations of folic acid,
B vitamins and zinc [35, 36].
Chapter 1. Background 5

Sugar Fortification

The most common micronutrient added to sugar is vitamin A, and fortification has been imple-
mented in numerous Central American countries [37]. Cold water soluble vitamin A palmitate
beadlets adhere to the surface of sugar crystals and are coated with a vegetable oil-antioxidant
mixture to form a premix. The vegetable oil fixes the palmitate beadlets onto the sugar crystals,
while the antioxidant prevents the oil from becoming rancid and enhances stability of vitamin
A palmitate. Lastly, the premix is blended with non-fortified sugar at 1:1000 ratio [38, 39].
A trial study conducted on iron fortified sugar resulted in increased iron stores in semirural
Guatemalan populations [40], but no industrial feasibility studies have been published at this
time.

Flour Fortification

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A staple food for many nations, flour is widely commercially fortified with an extensive range
of micronutrients added at varying levels [41]. WHO provides guidelines on the fortification
of flour with iron, folic acid, vitamin B12 , vitamin A and zinc [42]. In many countries, flour
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is enriched with B vitamins to compensate for micronutrient losses during the flour milling
process. A dry powder premix of micronutrients is added after the milling step and blended
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with flour to give fortified flour [43].

Oil Fortification
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Oil and its derivatives are ideal vehicles for the incorporation of fat-soluble micronutrients,
namely vitamins A, D, E and K [44]. Edible oils are primarily fortified with vitamin A and,
to a lesser extent, vitamin D due to nutritional needs and technical feasibility [44]. There have
been marked successes in the fortification of oil with vitamin A. Oil and vitamin A can be sim-
ply blended together without changing processing conditions, thus requiring minimal additional
costs and technical expertise [10]. Oxidative degradation reduces the stability of vitamin A in
oil, which could be countered by addition of antioxidants [44, 45]. Addition of minerals to oils
has also been attempted with limited success.

Rice Fortification

Similar to flour fortification, rice has also been fortified with a wide range of micronutrients
[46]; but unlike flour, preferences on the type of rice used and organoleptic features of cooked
rice differ significantly between cultures. This poses unique challenges in the standardization
of fortification processes [10]. WHO recommends the fortification of iron, vitamin A and folic
Chapter 1. Background 6

acid into rice [47]. In practice, rice is often enriched with B vitamins as they are lost during
rice milling processes that remove the outer germ and bran layers.
In rice fortification, fortified kernels are formed using coating or extrusion processes which
are subsequently blended with regular rice kernels at a 0.5-2% ratio [46].

1.3.2 Fortificants

As shown in subsection 1.3.1, the use of synthetic fortificants for fortifying staple food vehicles
is well studied. Pure micronutrients are used (with a food vehicle) to form a fortified premix
with a high concentration of fortificant. The premix is then blended with non-fortified version
of the food vehicle to give the appropriate level of fortification. A major advantage of using
synthesized fortificants is greater control over the level of fortification, provided that the mi-
cronutrient compounds are of high purity and the fortification process is designed and operated

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appropriately.
Many plants species contain a variety of micronutrients but have high moisture content,
making them extremely perishable and low in micronutrient content on a per volume basis.
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Plant species, especially green leafy vegetables, may also contain antinutritional compounds
such as oxalates, saponins and phytates that inhibit the absorption of nutrients for humans [48].
To increase the shelf-life, concentrations and bioavailability of the micronutrients, dehydration
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and blanching may be used to form “packages” containing micronutrients that could be used to
enrich food vehicles [48–51]. These “packages” are natural fortificants. In addition to increasing
the nutritional value of food vehicles, the components within the matrix of natural fortificants
may improve storage stability. Natural fortificants allow the micronutrients in plants to be
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readily available year-round and reduce wastage from food spoilage. Ranawana et al. [49] showed
that the fortification of bread using freeze-dried vegetable powders increased the nutritional
value and enhanced the storage stability of bread samples. Similarly, Duthie et al. [50] also
reported an increase in oxidative stability of meats with the addition of vegetable powders. On
the other hand, Joshi and Mathur [51] demonstrated that the sensory properties of Western
and Central Indian dishes remained acceptable when 10% w/w of vegetable leaf powders were
added to the recipes.

1.4 Moringa oleifera as a Natural Food Fortificant

Moringa oleifera is a plant species that grows abundantly in tropical and subtropical regions
and is one of 13 species classified under the Moringaceae family [52]. In this thesis, Moringa
oleifera is referred to as ‘Moringa’. The time period from seed sowing to the harvest of Moringa
fruits is six months, which is relatively short. Moringa fruits are also pods containing seeds
and are typically 20-75cm long and weigh 90-150g, while its leaves 25-45cm long and are made
oblique leaflets of 1cm in length. Moringa trees are also known as “drumstick trees” with
Chapter 1. Background 7

reference to the pods’ resemblance to the musical instrument. Moringa is a multifunctional

plant with uses in food, medicine [53], wastewater treatment [54, 55] and biofuels [56].
Various parts of Moringa are edible including its leaves and immature pods. India is the
largest producer of Moringa and cultivates trees spread over 38,000 hectares with an annual
production of Moringa pods of 1.1-1.3 million tonnes [57]. The production of Moringa leaves
vary greatly depending on cultivation conditions (e.g. tree spacing, weather, and varietal) and
yields up to 650 tonnes per hectare are found in literature [58]. Perennial Moringa trees require
close to one year of growth before first harvest and produce fewer pods in the first two years of
cultivation compared to annual Moringa trees (Table 1.3). Harvesting periods of Moringa are
constrained by environmental factors such as monsoons, which could cause flowering Moringa
trees to shed their blooms. A study conducted by the Tamil Nadu Agricultural University
showed that Moringa trees purposefully grown to yield harvest during conventional off-seasons
produced noticeably smaller and fewer pods [58].

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Table 1.3: Production yields and harvest periods for perennial and annual Moringa oleifera
Perennials Annuals
Production Yield First two years: 80-90
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(pods/year/tree) Year 4-5: 500-600
Harvest period(s) March-June (primary) six months after seed sowing;
(in tropical climates) September-October 2-3 months of harvest
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Moringa leaves are reported to be high in nutritional content [59] but have distinct astrin-
gent and grassy flavours, while immature Moringa pods are comparatively lower in nutritional
content [60] and have mild sweet and crisp flavours likened to asparagus. Moringa leaves is
presented in both academic and non-academic sources as containing more iron than spinach,
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more vitamin A than carrots, and more calcium than milk on an equivalent weight basis [61, 62].
A broad range of values exist in literature with regards to the nutritional content in Moringa
leaves, ranging from high to very high nutrition [52, 59–63]. Although variations are expected in
biological materials, the inconsistent values reported for Moringa on its micronutrient content
lead to uncertainties in the nutritional value of Moringa. Furthermore, despite growing interest
from health and agriculture communities on Moringa few regions currently include it as a reg-
ular food source. It is speculated that the astringent taste of Moringa leaves is not generally
pleasing and Moringa pods are less nutritious and require more effort to eat due its fibrous
outer skin. These challenges suggest that there are opportunities in increasing the consumption
of Moringa as a means to alleviate micronutrient deficiencies.
Given Moringa’s nutritive properties and abundance within the tropical region, which coin-
cides extraordinarily well with regions most burdened by food insecurity, a number of campaigns
have been launched to introduce Moringa as part of a diversification strategy. However, the
inclusion of new foods into people’s diets is often slow and challenging since it requires substan-
tial behaviour change. The diversification approach also does not address the key limitation of
the undesirable astringent or bitter taste in Moringa leaves. Consumption of fresh leaves and