FFN Compiled

Session 1
Introduction to
functional foods and
nutraceuticals
Junaida Astina, Ph.D.
FS3226-Functional Foods and Nutraceuticals
Course Learning Objectives (CLOs)
CLO1: Describe the definition, history, and global aspects of functional

foods and nutraceuticals
CLO2: Discuss the health aspects of various functional foods and
nutraceuticals
CLO3: Account for safety and efficacy of functional foods and
nutraceutical products
CLO4: Able to describe the role of functional food product for disease
prevention and health promotion
CLO5: Discuss regulatory issues in functional foods and nutraceuticals
products
2
Attendance
• Late 10-15 min tap-in allowed, seat in allowed

• Late >15 min tap-in not allowed, seat in allowed
• Minimum 80% attendance to join the final examination
• If you are sick and unable to come, send the medical certificate to my
email: junaida.astina@i3l.ac.id and cc to AO.
Course Outline
No. Topic
1 Introduction to functional foods and nutraceuticals
2 Classification of antioxidants
3 Prebiotics, probiotics and synbiotics - 1/2
4 Prebiotics, probiotics and synbiotics - 2/2
5 Carbohydrate-based functional foods
6 Lipids and lipid related functional foods
7 Marine bioactive compounds as functional food ingredients
8 Mid-term Examination
9 Proteins, peptides and amino acids as functional food ingredients
10 Vitamins and minerals as functional ingredients
11 Safety and efficacy of functional foods and nutraceuticals products
12 Health claims and labeling regulation
13 Consumer understanding of nutrition and health claims
14 Application of biotechnology and functional food development
15 Functional foods: opportunities, challenges and trends
16 Final-term Examination 4
Assignment and Quiz
• Two Quizzes
• Two Assignments: Indonesian Local Functional Food and/or
Nutraceuticals
• Make a review paper (1) session 7
• Create a functional food product development prototype session 15
• Both assignments should be in line, in terms of topic and content the
product category has been determined for each group.
• You are able to publish your review paper for scientific purposes, student
conferences, etc.
It s not mandatory, but there will be bonus points for final exam in this case
5
Assignment 1: Review Paper
• Review Papers
• More than literature overview critical and comprehensive reviews
provide new insights or interpretation of a subject through thorough and systematic evaluation of
available evidence
contain an in-depth critical review of the literature
• Purpose: writing practice and coaching

• First-come, first-served basis
Email me to set the schedule
Decide your team leader who will be the first author and manage the overall works. I will be the
corresponding author if it you intend to publish it.
• Max. 2 groups per session, max. 25 minutes/group use your time efficiently
• Format
• Follow the target journal, competition, etc.
• It can be local, national, or international-indexed journal different challenge
6
Assignment 1: review paper
(Not limited to) these are several points you can cover in the assignment:
• Description: about the product, traditional/local values/utilization
• Scientific approach:
• Bioactive compounds
• Potential health benefits
• Bioactivity and mechanism of actions
• Safety and efficacy in animal or human trials
• Potential technology/development can be applied to develop this product
• Future outlooks
7
Topics
Group Topic Members

1 Antioxidant Joya Angelica Immanuel
Valeska Refflyn
Antonia Caitlin Gondowardoyo
Graciella Virginia Irwantoho
Aqeela Syakira Sugarda
Novelin Rudijanto
Ashella Loral Lydia Claire
2 Probiotic/ probiotic/ synbiotic Vanessa Ellivia
Erika Marceline Kwan
Celestine Jovanka Tjahjadi
Andrea Benita Wiguna
Angelyna
Belinda Hariyanto
Chelsy Angelline Wen
Topics
Group Topic Members

3 Carbohydrate-based functional food Nathan Victor
Anjali Kothari
Janice Victoria Muliady
Gabrielle Maria Archangela
Rico Alexander Pratama
Vanessa Yulianti
Theo Millard Tirtawinata
4 Lipid-based functional food Alexandra Amanda Ningtyas
Clarabelin
Kim Jung Yeu
Aurelia Faustine Wu
Anganjani Samaraswasdee Arya
Willy Setiawan
Reynaldi Budiman W
Topics
Group Topic Members

5 Marine-based functional food Kayla Cleverie
Vania Abigail Pingadi
Arielle Praise Rachman
Vittorio Denovian
Jessy Julia Wie
Nadya Theodora
Keyzia Asarya
Review Paper: Grading Rubrics
• Writing organization (15%)

• Introduction (10%) Provide interesting insights regarding the problem
and demonstrate the importance of the topic
• Review content (60%)
• Student s thorough understanding of the topic
• State of the art (40%) State of art can be measured by demonstration of critical
thinking when discussing the study. It can be seen from: 1. the original opinion/
argument/ point of view of your group toward the objective, methodology, result,
discussion; 2. the significance point(s) that have been served in the paper; 3. see the
potential gap that probably exists between similar studies or toward what have been
studied in the class
• Similarity (max. 15%) (15%)
11
Assignment 2: Functional food product
development
• Purpose: boost creativity and apply your knowledge

• Use evidenced-based articles/journal as your main references (review
articles are allowed)
• We will have a mini exhibition about functional food product
development. You can present your product and poster/flyer which
include:
• Background
• Objective
• Materials and methodology
• Expected health benefits
• Packaging design of the product
• Poster should be saved as PDF file and uploaded to ACIS
12
Assessment
Type Weight *I will give peer evaluation (confidential) for each assignment,
therefore please have a fair and good team work!
Quiz 15%
** If you submit your review paper and get reviewed/
Assignments* 20% accepted/published/win competition, you will not only get the
score for the assignment portion, but also for the final exam as
Midterm Exam 30% the appreciation for your recognized works outside the classroom.
Final Exam** 35% Terms and conditions apply. You still need to do the exam. Later, we
will choose the best score from both.
Terms and conditions last submission of the proof: Final Exam Day
Scale Status/Condition Score for final exam
International Submitted to the journal/ conference/ competition 80

Under-reviewed (pass the editorial board review)/ Finalist 85
Accepted with major revision/ Top 5 Winner 90
Accepted with minor revision/ Accepted/ Published/ Top 3 Winner 100
Local/National Submitted to the journal/ conference/ competition 75
Under-reviewed (pass the editorial board review) 80
Accepted with major revision/ Finalist 85
Accepted with minor revision/ Top 5 Winner 90
Accepted/ Published/ Top 3 Winner 100
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Assessment: Overall
Score Range Grade Status

85 100 A Passed
80 84.9 A- Passed
75 79.9 B+ Passed
70 74.9 B Passed
65 69.9 B- Passed
60 64.9 C+ Passed
55 59.9 C Passed
50 54.9 D+ Failed
45 49.9 D Failed
0 44.9 E Failed
16
Functional food & autoxidation
Related learning outcomes
By the end of this session, students will be able to :

1. Accuracy in Accuracy in defining the terminology of functional food
and nutraceuticals
2. Accuracy in discussing the market trend of functional food and
nutraceuticals in Indonesia and abroad
3. Accuracy in describing biological antioxidants defense system
20
Course outline
Subtopics that will be delivered in this session:

1. Introduction
2. Definition of Functional Foods and Nutraceuticals
3. Search trends in Indonesia compared to survey results done in America and
Canada
4. Marketability
5. Antioxidants and antioxidants rich foods
• Autoxidation
6. Biological antioxidants defense system
21
Introduction
“Let the food be thy medicine and

medicine be thy food
1. Introduction
Food + Medicine
Functional Food
Minimum requirements for Food
PALATABILITY NUTRITIOUS NO DOSAGE

It has agreeable to It can provide you macronutrient, The limitation is the
the palate or taste. micronutrient, and non-nutrient amount you are able to eat.
components needed.
A food may have three functions:
1. Providing energy and other basic nutrition
2. Giving enjoyable aroma, color, and taste
3. Having health benefits
The utilization of food for preventing nutrient-deficiency diseases was

then becoming the starting-point of what we know now as healthy diet,
and thus acted as a trigger for the development and construction of the
terminology of functional food .
25
2. Definition of functional foods
• Functional foods are similar in appearance to conventional foods; the former

being consumed as part of the normal diet.
• In contrast to conventional foods, functional foods, however, have demonstrated
physiological benefits and can reduce the risk of chronic disease beyond basic
nutritional functions, including maintenance of gut health.
• The term functional food was coined by Japanese scientists in the s and was
introduced to the European scientific community in the s.
26
2. Definition of Nutraceuticals
• The term "nutraceutical" was coined from "nutrition" and "pharmaceutical" in

1989 by Stephen DeFelice, MD.
• DeFelice proceeded to define nutraceutical as, "a food (or part of a food) that
provides medical or health benefits, including the prevention and/or treatment
of a disease
27
Nutraceuticals
• Nutraceuticals differ from dietary supplements in the following aspects:

1. Nutraceuticals must not only supplement the diet but should also aid in the prevention
and/or treatment of disease and/or disorder; such as diabetes, atherosclerosis,
osteoporosis, cardiovascular disease, cancer and neurological diseases. Most of the
nutraceticals compounds have antioxidant activity.
2. Nutraceuticals are used as conventional foods or as sole items of a meal or diet.
• Some of the popular nutraceuticals such as Echinacea, glucosamine, omega-3,
lutein, and folic acid have been shown through scientific research to have
therapeutic properties
28
Indonesian Association of Functional Food and Nutraceuticals
29
Similar and differences between functional
food and nutraceutical
Similarity Differences
. “Search trends in Indonesia
compared to survey results
done in America and Japan
33
34
Current Status Of Functional Foods Research And Development
In Indonesia
Purwaningsih et al. 2021

DOI: 10.6066/jtip.2021.32.1.83 37
US
38
US
39
Canada
*In older adults

40
Canada
*In older adults

41
US
42
US
43
US
44
Functional food trend in Japan
What is your opinion
about this data?
47
4. Marketability
48
Functional food sales in Japan
Global trends of the functional foods market
Vicentini et al., 2016 51

Marketability: Functional Foods
Market share by key functional claims of
products 2013
(Source: Fuji Keizai Group, H.B Foods
Marketing Handbook 2014 Vol.3)
52
Marketability (segmentation): Supplements
Dietary supplements specifically intended help
meet recommended dietary requirements and
supplement regular diet (FMI, 2015)
Market Value and Forecast:
Estimated to be valued at US$ 123,324.7 Mn by the
end of 2015 and expected to account for US$
252,142.1 Mn by 2025 (FMI, 2015)
53
5. Antioxidants and
antioxidants rich foods
Focus: autoxidation
54
Oxidation - video
https://www.youtube.com/watch?v=p1PEWZRHylo 55
Oxidation
• Oxidation is one of the metabolic reactions in the body and in foodstuffs

essential for the survival of cells. Normal metabolism is dependent on
oxygen, a free radical. Through evolution, oxygen is thought of as the
terminal electron acceptor for respiration.
• Autoxidation is the direct reaction of molecular oxygen with organic
compounds under mild conditions.
• The dependence on oxygen for normal metabolism results in the
production of other oxygen-derived free radical species, such as
superoxide or hydroxyl radicals, formed during metabolism, energy
production in the body or by ionizing radiation.
• These oxygen-derived free radical species are stronger oxidants and are,
therefore, dangerous which cause oxidative damage leading to cell and
tissue injury.
• These free radicals are involved in both human health and disease.
56
Oxidation
• Free radicals are atoms or molecules having unpaired

electrons.
• The unpaired, or odd, electron is highly reactive as it
seeks to pair with another free electron.
• Free radicals are involved in enzyme-catalysed reactions,
electron transport in mitochondria, signal transduction
and gene expression, activation of nuclear transcription
factors, oxidative damage to molecules, cells and tissues,
antimicrobial action of neutrophils and macrophages,
aging and disease.
• When an excess of free radicals is formed, they can
overwhelm protective enzymes such as superoxide
dismutase, catalase and peroxidase and cause destructive
and lethal cellular effects (e.g., apoptosis) by oxidizing
membrane lipids, cellular proteins, DNA and enzymes,
thus shutting down cellular respiration. 57
Diseases related to oxidation
• Inflammation
• Auto immune disease
• Cardiovascular disease
• Aging
https://www.mdpi.com/2673-9801/2/2/6
Sources of free radicals
1. Normal aerobic respiration

2. Peroxisomes, which are organelles responsible for degrading fatty acids and
other molecules, produce H2O2 as a by-product, which is then degraded by
catalase. Under certain conditions, some of the peroxide escapes degradation,
resulting in its release into other compartments of the cell and in increased
oxidative DNA damage.
absorb light maximum 450nm
3. Cytochrome P-450 enzymes animal defense system against natural toxic
chemicals from plants
4. Phagocytic cells destroy bacteria or virus-infected cells oxidative burst: NO,
H2O2, etc.
59
Exogenous sources of oxidant
1. Cigarette smoking
2. Dietary factors: Iron and copper salts promote the generation of
oxidizing radicals from peroxides.
3. Radiation/UV light
4. Strenuous work or exercise, etc.
60
reactive oxygen species
reactive nitrogen species
https://www.mdpi.com/2673-9801/2/2/6
6. Biological antioxidants
defense system
62
too much free radicals —> our body will develop the disease 63
6. Biological antioxidants defense system
harmful
metabolism
vit E can be regenerated by vit C
in this picture they only mentioned the unsaturated fatty vit E can donate its
acid because the saturated fatty acid is double bond —> electron and
free radical can attack unsaturated fatty acid prevent free radical
vit A can also donate its electron and after that it 64

can be decayed will be decayed from our body
Discussion
1. Accuracy in Accuracy in defining the terminology of functional food

and nutraceuticals
- What is functional food, give examples?
- What is nutraceutical, give examples?
2. Accuracy in discussing the market trend of functional food and
nutraceuticals in Indonesia and abroad
- How is the market trend and future prospect in Indonesia?
- How is the market trend and future prospect in US?
- How is the market trend and future prospect in Japan?
65
Discussion

- What is autoxidation?
- What are the effects of oxidation on human health?
- What are the sources of oxidation?
- How does biological antioxidant system work?
66
References
• Chapter 1 and 2 Guo, M. (2009). Functional foods: principles and technology.

CRC press.
• Limón-Pacheco, J., & Gonsebatt, M. E. (2009). The role of antioxidants and
antioxidant-related enzymes in protective responses to environmentally induced
oxidative stress. Mutation Research/Genetic Toxicology and Environmental
Mutagenesis, 674(1), 137-147.
67
Session 2
Antioxidant
classification and
antioxidant-rich foods

1. Describe the basics of the classification of antioxidant
2. Mention and describe some of antioxidant rich foods
2
Course outline

1. Antioxidants
2. Natural antioxidants
3. Chemical classification of food antioxidants
4. Classification of antioxidants based on their function
5. Classification of antioxidants based on the site of synthesis
6. Foods rich in antioxidants
7. Role of antioxidant in disease prevention
3
• What is antioxidant?
• What does antioxidant do?
• What is the role of antioxidant in food?
• What is the effect of antioxidant in human body?
1. Antioxidants
• The onset of lipid oxidation can be delayed by adding
antioxidants to food or by including them in our diet.
• The major role of antioxidants in food industry is to
prevent off-flavors, rancidity and to maintain their
nutritional value.
• Food antioxidants are inhibitors of lipid peroxidation
and consequence food deterioration.
5
2. Antioxidants (cont’d)
• Hence, antioxidants are defined as

substances when present in foods
at low concentrations compared
with those of an oxidizable
substrate markedly delay or
prevent the oxidation of the
substrate (Halliwell, 1999)
6
Defining antioxidants
• The term “antioxidant” has many definitions

• Chemical definition: “a substance that opposes oxidation or inhibits reactions
promoted by oxygen or peroxides”
• Biological definition: “synthetic or natural substances that prevent or delay
deterioration of a product, or are capable of counteracting the damaging effects
of oxidation in animal tissues”
• Institute of Medicine definition: “a substance that significantly decreases the
adverse effects of reactive species such as ROS or RNS on normal physiological
function in humans reactive oxygen species
reactive nitogen species
Huang, et al, J. Agric. Food Chem. 2005, 53: 1841-1856

• On the other hand, in the human gastrointestinal tract and within the body
tissues, oxidative damage to proteins and DNA is as important as damage to
lipids.
• Oxidative DNA damage could be a major risk factor for the development of
tumors, so that dietary antioxidants able to decrease such damage in vivo would
be expected to have cancer prevention effects.
8
• The term oxidizable substrate encompasses almost everything (except water)

found in foods and in living tissues and includes proteins, lipids, carbohydrates
and DNA molecules.
• An antioxidant may be able to protect one biological or food system but it may
fail to do so in others.
• For example, antioxidant inhibitors of lipid peroxidation may not protect other
molecular targets such as DNA and protein against oxidative damage and may
sometimes aggravate such damage.
9
Video - antioxidant
https://www.youtube.com/watch?v=KCF6prDSrHE
10
• Antioxidants have become an indispensable group of food additives. The use of

antioxidants dates back to the 1940s.
• Gum guaiac was the first antioxidant approved for the stabilization of animal fats,
especially lard.
• The resin from the wood of Guajacum officinale L., or of Guajacum sanctum L.,
(Fam.Zygophyllaceae), consisting of approximately 70% alpha- and beta-
guaiaconic acids, 10% guaiaretic acid, and 15% guaiac beta-resin and small
quantities of guaiac yellow, vanillin, etc.
11
Guaiacum officinale, commonly known as roughbark lignum-vitae,
guaiacwood or gaïacwood, is a species of tree in the caltrop family,
Zygophyllaceae, that is native to the Caribbean and the northern
coast of South America.
12
2. Natural antioxidants
Antioxidants in food are important for four reasons:

• First, endogenous antioxidants may protect components of the food itself against
oxidative damage.
• For example, spices rich in antioxidants have been used for centuries to delay oxidative deterioration of
foods during storage or cooking.
• Second, dietary antioxidants may be absorbed into the human body and exert
beneficial effects.
• For example, quercetin and catechins can be absorbed to some extent in humans and they and their
metabolites can reach plasma concentrations in the range of 0.1 – 1 μM. Such concentrations can, in
vitro, delay the process of lipid peroxidation in liposomes, microsomes and low-density lipoproteins
(LDL).
Oxidized LDL (oxLDL) represents a variety of modification of both lipid and apolipoprotein B (apoB) components by lipid
peroxidation. This promotes atherosclerosis through inflammatory and immunologic mechanisms that lead to the
formation of macrophage foam cells.
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2. Natural antioxidants (cont’d)
• Third, food derived antioxidants could exert beneficial effects, without being
absorbed, in the gastrointestinal tract itself.
• Fourth, there is great interest in plant extracts for therapeutic use as anti-
inflammatory, anti-ischemic, and antithrombotic agents.
• An extract of the ornamental tree Ginkgo biloba has been used in herbal
medicine for thousands of years: the extract has antioxidant properties in vitro,
apparently largely from the flavonoids present, which include rutin, kaempferol,
quercetin, and myricetin.
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Ginkgo biloba
15
Fig. 1. Influence of butylated hydroxytoluene (BHT) or ethoxyquin (EQ) and of ethyl ether extracts (EEE), ethyl acetate
extracts (EAE), acetone extracts (AE), ethanol extracts (EE) or aqueous extracts (AQE) of Ginkgo biloba leaves on
absorbance value at 500 nm for the FTC method (A) and at 532 nm for the TBA method (B). The inhibition (I) induced by these
compounds on the lipid oxidation measured using the FTC method (C) and the TBA method (D) in a linoleic acid emulsion. The data
represent the means ± SD of 4 replicates.a–d Bars with different superscripts are significantly different (P < 0.05).
Huatao Li, Xiaoqiu Zhou, Ping Gao, Qiuyue Li, Hansi Li, Rong Huang, Min Wu
Inhibition of lipid oxidation in foods and feeds and hydroxyl radical-treated fish erythrocytes: A comparative study of
Ginkgo biloba leaves extracts and synthetic antioxidants
Animal Nutrition, 2016, Available online 29 April 2016
http://dx.doi.org/10.1016/j.aninu.2016.04.007
2. Natural antioxidants (cont’d)
• Of the natural antioxidants, two important groups, the tocopherols and ascorbic
acid, are highly effective in many food products.
• As an example, oat and amaranth oils contain high levels of antioxidants such as
tocopherols and squalene. These oils might be added to certain other oils in
order to stabilize them.
• Furthermore, extracts of green tea, rosemary and sage might be used in a variety
of foods in order to control oxidation.
17
3. Classification of natural antioxidant
Carocho & Ferreira. 2013. Food and Chemical Toxicology 51 (2013) 15–25
4. Classification of antioxidants based on their
function
1. Primary or chain breaking antioxidants (scavenger antioxidants):

• These antioxidants can neutralize free radicals by donating one of their own
electrons, ending the electron “stealing” reaction.
• The resultant antioxidants which become free radicals, because of one
electron left in their outer shell, are relatively safe, stable and in normal
circumstances insufficiently reactive to initiate any toxic effect, e.g., α-
tocopherol.
21
The in vitro antioxidant activity of tocopherol. FRS,
free radical scavenging activity; LP, lipid peroxidation;
SHP, scavenging of hydrogen peroxide; SOA,
superoxide anion scavenging activity. Values are
expressed as the mean±SEM.
Ilavenil et al., 2011.
22
function (cont’d)
2. Secondary or preventive antioxidants. These antioxidants act

through numerous possible mechanisms like:
a. Sequestration (isolation) of transition metal ions which are not allowed to
participate in metal catalyzed reactions;
b. removal of peroxides by catalases and glutathione peroxidase, that can
react with transition metal ions to produce ROS;
c. removal of ROS, etc.
23
• These (secondary or preventive) antioxidants which are also called as synergistic
antioxidants can be broadly classified as oxygen scavengers and chelators.
• Oxygen scavengers such as ascorbic acid, ascorbyl palmitate, sulfites and
erythorbates react with free oxygen to remove it in a closed system.
• Chelators like ethylenediaminetetraacetic acid (EDTA), citric acid, and phosphates
are not antioxidants, but they are highly effective as synergists with both primary
antioxidants and oxygen scavengers.
24
function (cont’d)
3. Tertiary antioxidants: These antioxidants remove damaged

biomolecules before they can accumulate and before their presence
results in altered cell metabolism and viability.
• For example, methionine sulphaoxide reductase repairs damaged DNA,
proteolytic enzyme system remove oxidized proteins and lipases, peroxidases
and acyl transferases act on oxidized lipids.
25
5. Classification of antioxidants based on the site of
synthesis
• Some antioxidants are synthesized within the cells themselves which are called as
endogenous antioxidants and others are found in food referred to as natural
antioxidants (Figure 2.5).
26
6. Foods rich in antioxidants
• Berries
• Grapes
• Tomatoes
• Garlics
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Berries
28
Berries (cont’d)
• Small berries constitute an important source of potential health promoting

phytochemicals (such as flavonoids). These include fruits of the Vaccinium, Rubes,
Ribus and Fragaria genera.
• Examples of Vaccinium genus are lowbush and highbush blueberry, bilberry,
cranberry and lingonberry.
• Examples of Rubus genus are blackberries, red and black raspberries.
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Cranberries
• The fruits of American cranberries, Vaccinium macrocarpon and European

cranberries, Vaccinium oxycoccus have been associated with a variety of health
benefits.
• There are reports of its use by American Indians to dress wounds and prevent
inflammation.
• In the early 20th century, cranberries were thought to help relieve the symptoms
of urinary tract infections, or perhaps even prevent their occurrence.
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Chemical composition of cranberries
• Proanthocyanidins and anthocyanins make up the pigment of the leaves, and

produce the color of the berries.
• More importantly, proanthocyanidins are responsible for the cranberry’s best-known
medicinal effect, preventing bladder and urinary tract infections by inhibiting bacterial
colonization. They may also help relieve diarrheal symptoms.
• Organic acids, including quinic, malic, and citric acids.
• Quinic acid is considered the most important among these organic acids. These compounds,
which are responsible for the sour taste of cranberries, acidify the urine and prevent kidney
stones.
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Chemical composition of cranberries
• Vitamins and minerals. Cranberries are rich sources of vitamins including vitamin
A, carotene, thiamine, riboflavin, niacin, and vitamin C.
• They also contain many essential minerals such as sodium, potassium, calcium,
magnesium, phosphorus, copper, sulfur, iron, and iodide.
• These vitamins are strong antioxidants that enable cranberries to help protect
the body against such infections as colds or influenza.
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Antioxidants in cranberries
• Cranberry fruits serve as an excellent source of anthocyanins, flavonol glycosides,

proanthocyanindins and phenolic acids.
• Cranberries contain about 1g/kg of fresh weight of phenolic acids predominantly
as glycosides and esters.
• Twelve phenolic acids have been identified in cranberries (Figure 2.6).
• Sinapic, caffeic and p- coumaric acids are the most abundant bound phenolic
acids and coumaric, 2,4dihydroxybenzoic and vanillic acids the predominant free
phenolic acids found in cranberry.
• Resveratrol (0.25 mg/kg) has also been detected in cranberry fruit.
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34
35
FIGURE 5. Mechanism of SIRT1
activation by resveratrol. The effect
of resveratrol is similar to effects
observed upon low caloric diet, it
also leads to increase in cAMP.
Resveratrol inhibits PDEs (phospho

diesterase), leading to increased
cAMP (cyclic AMP) levels, Epac1
(cAMP-regulated guanine nucleotide
exchange factor 1) activation,
elevated intracellular calcium, and
AMPK activation.
Downstream of AMPK, an increase in

NAD + levels leads to SIRT1
Gerszon, J., Rodacka, A., & Puchała, M. (2014). activation, which promotes
Antioxidant properties of resveratrol and its
beneficial metabolic changes
protective effects in neurodegenerative
diseases. Advances in Cell Biology, 4(2), 97-
primarily through deacetylation and
117. activation of PGC-1α.
36
Therapeutic effects of cranberries
• Urinary tract infections (UTIs)

• Oral cavity health
• Other benefits
37
Urinary tract infections (UTIs) treatment -
cranberries
• Earlier it was thought that cranberry’s effect on acidification of the urine as the
possible mechanism for cranberry’s antibacterial effect in the urinary tract, but
this theory was not substantiated by other research (Avorn et al., 1994).
• For UTIs to occur, bacterial entry and proliferation must occur. Proliferation
requires attachment to urinary tract mucosal surfaces.
• The latest research supports the hypothesis that cranberry juice acts to promote
urinary tract health by inhibiting bacterial adherence to mucosal surfaces (Henig
and Leahy, 2000; Leahy et al., 2001).
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Urinary tract infections (UTIs) treatment -
cranberries
• Recently, a study was completed in which extracts of purified cranberry

proanthocyanidines (PACs) were fed to mice.
• The urine was found to exhibit anti-adhesion activity against P-fimbriated E. coli,
providing the first in vivo evidence that cranberry PACs and/or metabolites can be
absorbed into the blood, and into urine, thereby eliciting this anti-adhesion effect
(Howell et al., 2001).
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Oral cavity health - cranberries
• Various bacteria appear to be major causative factors in the etiology of both

dental caries and periodontal gum disease.
• Of the hundreds of bacteria and bacterial pairs that could comprise the dental
plaque, Weiss et al (1997) isolated a wide variety of bacteria from the human
gingival crevice, and used a coaggregation assay to measure both aggregation and
the reversal of aggregation in the presence and absence of a selected cranberry
fraction in vitro.
• The isolated cranberry fraction was able to inhibit the coaggregation of 70% of
the bacterial pairs tested when at least one was Gram negative
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Other benefits - cranberries
• In vitro and in vivo animal studies have found anti-inflammatory, anticarcinogenic,

antiplatelet aggregation, vasodilatory, and other effects of several of antioxidant
compounds.
• Preliminary research on various proanthocyanidins suggests that they may act as
antioxidants, and have cardioprotective and anticarcinogenic activities (Ho et al.,
1999).
• Phenolic acids may contain antibacterial, antifungal, anticancer effects and
activity.
• Ellagic acid has been shown to have a broad range of anticarcinogenic activity.
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Grapes
42
Grapes
• Grapes belonging to the species Vitis vinifera L. are predominantly cultivated in

Europe, while those belonging to species Vitis labrusca and Vitis rotundifolia are
grown in North America (Mazza, 1995).
• Approximately 80% of the total crop is utilized for wine making; 13% is consumed
as table grapes and 7% processed into juice and raisins.
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Antioxidants in grapes
• Anthocyanins are the predominant phenolics of red table grape varieties, while
flavanols are the main phenolics in white table grape varieties (Cantos et al.,
2002).
• Grape seeds, stems and skins are a rich source of health-promoting flavonoids
such as proanthocyanidins, flavonols and flavan-3-ols.
• Proanthocyanidins are the major polyphenols in grape seeds, stems and skins.
• Procyanidins are the predominant proanthocyanidins in grape seeds, while
• Procyanidins and prodelphinidins are dominant in grape skins and stems
(Souquet et al., 2000)
44
45
46
Therapeutic effects of grapes
• Antioxidants in grapes are believed to protect the body from certain cancers and
heart disease.
• Resveratrol has anti-infective, anti-inflammatory and antioxidant properties in
humans. This compound helps battle cancer in various stages, from initiation to
promotion to progression.
• Grapes’ antioxidant properties have been shown to strengthen blood vessels,
boost immunity and inhibit allergies.
47
Tomatoes
48
Tomatoes
• The tomato (Lycopersicon esculentum) is one of the world’s major vegetables

with 4.4 million hectares under production and 115 million tons produced
worldwide in 2004 (FAOSTAT, 2004).
• Americans each eat more than 16 pounds of fresh tomatoes a year and consume
the equivalent of 79 pounds in processed tomatoes annually.
• Tomatoes are a rich source of antioxidants, including vitamin C and lycopene.
49
Antioxidants in tomato
• Lycopene, the carotenoid pigment responsible for the red color, is the most
distinctive compound present in tomatoes and has been recognized as the most
effective antioxidant among the carotenoids.
• The total flavonol content of tomatoes grown in different countries ranges from
1.3 to 36.4 mg of quercetin/kg of fresh weight (Dewanto et al., 2002).
• Quercetin conjugates are the predominant form of flavonols found in tomatoes,
but smaller quantities of kaempferol conjugates and traces of free aglycons have
also been detected.
50
Lycopene
51
Therapeutic effects of tomatoes
• Lycopene and β-carotene have been shown to act as powerful antioxidants in

humans.
• A diet containing moderate amounts of lycopene has been associated with the
prevention of cardiovascular disease and cancers of the prostate and
gastrointestinal tract (Rao and Agarwal, 2000).
• One 6-year, prospective, epidemiological study of approximately 47,000 men, the
Health Professional Follow-up Study (HPFS), concluded that 2 to 4 servings per
week of raw tomatoes significantly reduced the risk of prostate cancer by 26%
compared to no servings per week.
52
Garlics
53
Garlics
• Garlic has been called Russian penicillin. It belongs to the Lily family.
• Garlic is not just spice, herb or vegetable but a combination of all the three.
• Americans consume <0.6 g/week compared with 16 g/day in parts of China with
no side effects.
54
Antioxidants found in garlic
• Garlic has many functions. Some of sulfur containing compounds in garlic are E-
ajoene, Z-ajoene, allicin, allixin, allyl mercaptan, allyl methyl sulfide, diallyl
disulfide, diallyl sulfide, diallyl trisulfide, s-allyl cysteine and s-
allylmercaptocysteine.
• Among these, the chief active ingredient is allicin.
55
56
Therapeutic effects of garlics
• Research on the health benefits of garlic has tended to focus on fresh garlic
powders, aged garlic extracts, distilled oils (from steaming), and other forms.
• Research studies on garlic cover a wide topical area, from cardiovascular benefits
to garlic’s anti-carcinogenic properties.
• Garlic’s benefits include the reduction of harmful LDL cholesterol and
triglycerides;
• the prevention of blood clots and blood platelet clumping;
• the ability to block chemical carcinogens;
• The stimulation of various immunological factors that may help combat cancer as well as
fungal infections; and
• the protection of cells against various oxidizing agents.
57
• The free radical reactions which involve biological molecules (DNA, protein, and
lipids) appear to occur constantly → consequence of aerobic environment.
• To adapt with this condition, cells have developed system of defenses to prevent
and repair the injury associated with the oxidative changes to the biological
molecules → superoxide dismutases, glutathione system, vit E, ascorbic acid,
DNA repair enzyme, proteases to degrade damaged proteins, etc.
https://jcp.bmj.com/content/54/3/176
58
59
• It is only when the homeostatic mechanisms fail to keep pace with those
reactions that detrimental effects become evident.
60
Possible free radicals related diseases
62
• Ischemia-reperfusion injury and
stroke are associated with production
of superoxide (O2dot-) by the
mitochondrial electron transport
chain complexes I (NADH:ubiquinone
oxidoreductase) and III (cytochrome
C–coenzyme Q oxidoreductase).
• Superoxide dismutase forms
hydrogen peroxide (H2O2) from
superoxide. In the presence of
catalytically active metals, such as
Fe2+ or Cu+, hydrogen peroxide
breaks down to hydroxyl radical
(dotOH).
• Hydrogen gas (H2) reduces dotOH to
H2O, thereby preventing the lipid
peroxidation, DNA oxidation, thiyl
The hydrogen highway to reperfusion
radical formation and mitochondrial
therapy
depolarization that contribute to
Katherine C Wood & Mark T Gladwin
cellular apoptosis and necrosis.
Nature Medicine 13, 673 - 674 (2007) cell death
doi:10.1038/nm0607-673
63
64
Possible free radicals related diseases
65
References
• Chapter 1 and 2 – Guo, M. (2009). Functional foods: principles and technology. CRC press.
• Limón-Pacheco, J., & Gonsebatt, M. E. (2009). The role of antioxidants and antioxidant-related
enzymes in protective responses to environmentally induced oxidative stress. Mutation
Research/Genetic Toxicology and Environmental Mutagenesis, 674(1), 137-147.
• Hilgendorff, A., & O’Reilly, M. A. (2015). Bronchopulmonary dysplasia early changes leading to
long-term consequences. Frontiers in medicine, 2, 2.
• Ilavenil, S., Kaleeswaran, B., Sumitha, P., Tamilvendan, D., & Ravikumar, S. (2011). Protection of
human erythrocyte using Crinumasiaticum extract and lycorine from oxidative damage induced by
2-amidinopropane. Saudi journal of biological sciences, 18(2), 181-187.
67
Session 3
Prebiotics, probiotics,
and synbiotics
(part 1)
In-class activity
• Bring to campus: 1 product that you think contains

prebiotics/probiotics/synbiotics
• It can be natural/fresh food or processed food

1. Explain the basics of prebiotics, probiotics and synbiotics
2. Describe the effects of prebiotics and probiotics on various
physiological functions
3
Course outline

1. History of health claims concerning living microorganisms in food
2. History of the term probiotics
3. Definition of prebiotics
4. Definition of synbiotics
5. Pro-and prebiotics as functional foods
6. Synbiotics, probiotics or prebiotics in infant formula
7. Inulin
4
Can you mention foods/beverages containing
prebiotics/probiotics/synbiotics?
1. History of health claims concerning living
microorganisms in food
• There is a long history of health claims concerning living microorganisms in food,

particularly lactic acid bacteria.
• Since 7000 BC, archaelogists reported

the use of microorganisms in the
production of fermented beverages.
• Beer was produced from rice, honey,
and fruit with yeast fermentation
• Bread was used as yeast starter and
added to hulled grains and kept in
fermentation tanks
6
microorganisms in food - cont’d
• Since the advent of the microbiology era, some
investigators [eg, Carre (1887 – in German), Tissier
(1984), and Metchnikoff (1907)] attributed such health
effects to shifts of the intestinal microbial balance.
• Metchnikoff claimed that the intake of yogurt containing
lactobacilli results in a reduction of toxin-producing
bacteria in the gut and that this increases the longevity
of the host.
Élie Metchnikoff, Russian in
full Ilya Ilich Mechnikov • Tissier recommended the administration of
bifidobacteria to infants suffering from diarrhea,
claiming that bifidobacteria kills the pathogenic bacteria.
He showed that bifidobacteria were predominant in the
gut flora of breast-fed infants.
7
Sherman, 2011
9
microorganisms in food - cont’d
• The significant role of the intestinal microflora for resistance to disease was
shown by Bohnhoff et al (1954), Freter (1954, 1955, 1956), and Collins and Carter
(1978).
• Oral administration of antibiotics to mice rendered the animals more susceptible
to infection with Salmonella typhimurium, Shigella flexneri, and Vibrio cholerae.
• ≤1×101 Salmonellae enteritidis were sufficient to kill germ-free guinea pigs,
whereas 1×109 bacteria were required to kill animals with complete intestinal
microflora.
10
2. History of the term probiotics
In 1953, Kollath introduced Probiotic from the Latin, pro and the Greek β which
means “for life” → Active substances that are essential for a healthy development of life.
In 1965, Lilly and Stillwell defined probiotics as “substances secreted by one organism
which stimulate the growth of another”
In 1992, Fuller defined probiotics as “a live microbial feed supplement which beneficially
affects the host animal by improving its intestinal microbial balance
FAO/WHO guideline for probiotic evaluation
Characteristic of an ideal probiotic strain
• FAO and WHO set the

guideline for effective
evaluation of probiotics in
foods to substantiate the
health claims and benefits:
1. Strain identification
2. Functional characterization of
the strain(s) for safety and
probiotic attributes
3. Validation of health and
benefits in human studies
4. Honest, not misleading
labelling of efficacy claims and
content for the entire shelf
life.
Pandey, Kavita R., Suresh R. Naik, and Babu V. Vakil. Journal of food science and technology 52 (2015): 7577-7587.
Some approved probiotics:
• Lactobacillus rhamnosus
• Lactobacillus reuteri
• Bifidobacteria
• Certain strains of
Lactobacillus casei
• Lactobacillus acidophilus-
group
• Bacillus coagulans
• Escherichia coli strain
Nissle 1917
• certain enterococci,
especially Enterococcus
faeciumSF68
• yeast Saccharomyces
boulardii.
3. Definition of Prebiotics
• The term prebiotic was introduced by Gibson and Roberfroid (1995)
who exchanged “pro” for “pre,” which means “before” or “for.”
• They defined prebiotics as “a non-digestible food ingredient that
beneficially affects the host by selectively stimulating the growth
and/or activity of one or a limited number of bacteria in the colon.”
• This definition more or less overlaps with the definition of dietary
fiber, with the exception of its selectivity for certain species.
prebiotic can be fibre but not all of the fibre are prebiotic
• This selectivity was shown for bifidobacteria, which may be promoted
by the ingestion of substances such as fructooligosaccharides and
inulin, transgalactosylated oligosaccharides, and soybean
oligosaccharides.
14
Source of
prebiotics
• Source of prebiotics:
• Breast milk → direct
breastfeeding has greater
benefits
• Soybean
• Inulin (from Jerusalem
artichoke, chicory roots,
etc)
• Raw oats
• Unrefined wheat
• Unrefined barley & yacon
• Non-digestible carbs
Ideal properties of prebiotics
An ideal prebiotic should be:

1. Resistant to the actions of acids in
the stomach, bile salts and other
hydrolyzing enzymes in the intestine
2. Should not be absorbed in the upper
gastrointestinal tract.
3. Be easily fermentable by the
beneficial intestinal microflora
the higher DE (dextrose equivalent, the sweeter the product)
• The term synbiotic is used when a product contains
both probiotics and prebiotics.
• Because the word alludes to synergism, this term
should be reserved for products in which the
prebiotic compound selectively favors the probiotic
compound.
• However, one might argue that synergism is attained
4. Definition of in vivo by ingestion of lactobacilli on the one hand
synbiotics and promotion of indigenous bifidobacteria on the
other hand.
• Major probiotic strains in synbiotics formulations:
Lacbobacilli, Bifidobacteria spp, S. boulardii, B.
coagulans etc., while the major prebiotics used
comprise of oligosaccharides →
fructooligosaccharide (FOS), GOS and
xyloseoligosaccharide (XOS), inulin.
17
Novel prebiotics and probiotics
https://www.youtube.com/watch?v=0z47wLZ4-O4
5. Pro- and prebiotics as functional foods
• Among the most promising targets for functional foods are the gastrointestinal
functions, including those that control transit time, bowel habits, and mucosal
motility as well as those that modulate epithelial cell proliferation.
poor absorption, kebanyakan BAB
• Promising targets are also gastrointestinal functions that are associated with a
balanced colonic microflora → control of nutrient bioavailability, that modify
gastrointestinal immune activity, or that are mediated by the endocrine activity of
the gastrointestinal system.
• Bacteriotherapy is an alternative and promising way to combat infections by
using harmless bacteria to displace pathogenic microorganism.
21
Intestinal flora
22
Intestinal flora
• The intestinal flora has also been found to improve resistance to the colonization
by enteropathogenic micro-organisms such as Salmonella.
• The intestinal flora aids the digestion of food components, in particular of poorly
digestible carbohydrates. The butyric acid formed in that process has a favorable
effect on the intestinal epithelium.
• The intestinal flora also plays a role in fat metabolism (bile acid and cholesterol
metabolism) and the synthesis of vitamin K.
23
24
Health benefits of bifidobacteria
• Bifidobacterium is a non-spore-forming, Gram-positive, anaerobic probiotic

actinobacterium
• Bifidobacterium is commonly found in the gut of infants and the uterine region of
pregnant mothers.
• Bifidobacteria confer health benefits on the host when administered in adequate
amounts, showing multifaceted probiotic effects. For examples B. bifidum, B.
breve, and B. longum, common Bifidobacterium strains employed to prevent and
treat gastrointestinal disorders.
• Probiotics have been widely used in various functional foods, e.g., yoghurt, milk,
cheese, infant formula, and dietary supplements. The most common probiotics
include Lactobacilli and Bifidobacteria, which predominantly inhabit the animal
or human intestinal tract.
Chen, J., Chen, X., & Ho, C. L. (2021). Frontiers in Bioengineering and Biotechnology, 9, 1371.
25
Health benefits of bifidobacteria
Chen, J., Chen, X., & Ho, C. L. (2021). Frontiers in Bioengineering and Biotechnology, 9, 1371.
Health benefits of probiotics
• Probiotic-pathogen interactions in the middle part of

the figure include three mechanisms:
(1) direct combination,
(2) competitive exclusion,
(3) secretion of antimicrobial compounds
• Probiotic-host interactions in the right part of the
figure include three mechanisms:
(4) synergistic effects with indigenous microbiota,
(5) enhancement of epithelial barrier integrity, (6)
modulation of immune system.
• At the intestinal level in the left part of the figure,
probiotics have an effect through:
(7) upregulation of electrolyte absorption,
(8) modulation of gut motility,
(9) alteration of painful sensations.
Ma, L., Tu, H., & Chen, T. (2023). Nutrients, 15(2), 291.
Health benefits of prebiotics
Prebiotics consumed are fermented by gut microbiota and produce short-chain fatty acids (SCFA), such as
acetate, propionate, and butyrate. Those SCFAs promotes several health benefits for local, systemic, and
epigenetic modifications
Neri-Numa, I. A., & Pastore, G. M. (2020). Food Research International, 131, 108973.
6. Synbiotics, probiotics or prebiotics in infant
formula
30
31
Fructans
• As we know, all prebiotics to date have been carbohydrates, ranging from small
sugar alcohols and disaccharides, to oligosaccharides and large polysaccharides.
• Fructans are polymers of fructose that often contain a terminal glucose moiety.
They can have a linear or branched structure.
• Inulin-type fructans consist of fructose units linked through β-2,1 bonds and
displaying a linear structure.
• Fructan is also known as polyfructosylfructose.
32
33
7. Inulin
• The use of miscellaneous fructan- (but mostly inulin-) containing

plants as food seems to be quite old, dating back to at least 5000
years.
• In the early 1800s, a German scientist named Valentine Rose
discovered inulin from the boiling-water extract of the roots of
Elecampane (Inula helenium) and was later on named inulin by
Thomson in 1817.
34
Elecampane (Inula helenium)
35
7. Inulin
• Inulin belongs to a class of dietary fibers known as fructans.

• It consists of chain-terminating glucosyl moieties and a
repetitive fructosyl moiety, which are linked by β(2,1) bonds.
36
37
Chicory
38
Gembili
39
• In the classical purification process in order
to remove impurities from the extracted
juice, clarification requires multiple steps
(pre-liming, liming and carbonation) at
relatively high temperature (80-90°C) as
shown in Fig. 2 (Franck & De Leenheer,
2005).
• This may lead to the hydrolysis of inulin
molecules in the extracted juice and may
also introduce additional calcium ions into
clarified juice which requires further
purification treatments (Kim, Faqih & Wang,
2001).
40
Health benefits of inulin
• Almost 90 % of the inulin passes to the colon and digested by bacteria present
there (Cherbut, 2002). It is thus a significant component of the dietary fiber
complex and is labeled as dietary fiber on food items.
• Letexier and coworkers concluded from a study that incorporation of high-
performance inulin i.e., 10 g/d to a carbohydrate rich and reduced lipid food
exerted positive outcome on plasma lipids of human by lowering blood
lipogenesis and plasma triacylglycerol concentrations thereby reducing the risk
of atherosclerosis (Letexier, Diraison & Beylot, 2003)
41
Health benefits of inulin
• Owing to prebiotic action of inulin, it acts as a substrate for probiotics.

Chicory inulin (20-40 g/day) had a sufficient cathartic result in relieving
constipation (Fernández-Bañares, 2006; Kolida & Gibson, 2007).
• Oliveira et al. (2011) determined that the addition of inulin at a low
concentration in low fat milk has considerably enhanced the growth and
sustainability of Lactobacillus acidophilus, Lactobacillus rhamnosus and
Bifidobacterium lactis.
42
Types of prebiotics other than inulin and
fructooligosaccharides (FOS)
• Fiber gums; often used in such foods as yoghurt to give the product a thicker
consistency.
• Fiber gums are water-soluble and derived from such plants as acacia,
carrageenan, guar, locust bean, and xanthan.
• Usually containing about 85% fiber, these gums help promote the production of
large quantities of short-chain fatty acids, which are known to play several
beneficial roles, including the development of such intestinal bacteria as
Lactobacillus and Bifidobacteria.
43
Types of prebiotics other than inulin and
fructooligosaccharides (FOS)
• Lactulose; is a semisynthetic disaccharide comprised of galactose and fructose.

• Lactulose is resistant to human digestive enzymes and can be fermented by a
limited number of bacteria in the colon, especially Lactobacilli and Bifidobacteria.
• In the US, it is used as drug for treatment of constipation, but it has not been
proven to be a prebiotic substances.
• In Japan, it is marketed as a dietary supplement and used in functional foods.
44
Application of synbiotics in food industry
PROBIOTICS
PREBIOTICS
• Inulin was less sweet than sugar (sucrose) and was

moderately soluble in water, colourless, and clear to hazy,
with a pH of 5.84.
• The addition of inulin at 1% enhanced the microstructural,
textural and sensory properties of yoghurt compared to those
of the control
• This functional synbiotic product contains both probiotics (>6
log cfu g−1) and prebiotics.
• The inulin addition to the probiotic yoghurt improved the
viability of S. thermophilus, L. bulgaricus, L. acidophilus and B.
More than just a yoghurt! bifidum during storage.
The sensory evaluation showed that 1%
addition of inulin to low-fat probiotic
yoghurt had a good acceptability, similar
to full-fat probiotic yoghurt.
Discussion
Claimed by the company

Belligo is a probiotic on-the-go snack. Easy bite format has a
well-developed technology to ensure that the probiotics reach
the intestine safely without getting destroyed in the digestive
tract or by external factors such as heat or extreme
temperatures. To help these probiotics, Belligo also has
prebiotics that acts as food for the good bacteria. Prebiotic and
Probiotic together help in supporting digestion and immunity.
Belligo is made with natural ingredients that have a delightful
note of roasted coconut flakes.
What should be considered by the consumers?

Reading
Cruz, Adriano G., et al. "Sensory analysis: relevance for prebiotic, probiotic, and synbiotic
product development." Comprehensive reviews in food science and food safety 9.4
(2010): 358-373.
49
Short Sharing Session
• Local functional food and nutraceutical ideas – Topic and Outline
50
References
• Chapter 4 – Guo, M. (2009). Functional foods: principles and technology. CRC

press.
• Schrezenmeir, J., & de Vrese, M. (2001). Probiotics, prebiotics, and synbiotics—
approaching a definition. The American journal of clinical nutrition, 73(2), 361s-
364s.
• Ishizuka, T., Kanmani, P., Kobayashi, H., Miyazaki, A., Soma, J., Suda, Y., ... & Saito,
T. (2016). Immunobiotic Bifidobacteria Strains Modulate Rotavirus Immune
Response in Porcine Intestinal Epitheliocytes via Pattern Recognition Receptor
Signaling. PloS one, 11(3), e0152416.
51
Session 4
Prebiotics, probiotics,
and synbiotics
(part 2)

1. Explain about probiotic yogurt, cheese, and ice cream
2. Describe some health benefits of probiotic yogurt, cheese, and ice cream
3. Explain health benefits of probiotics, prebiotics, and synbiotics (gut-brain axis)
2
Course outline

1. Probiotic Yogurt
2. Probiotic Cheese
3. Probiotic Ice Cream
4. Benefits of probiotics, prebiotics, and synbiotics (gut-brain axis)
3
1. Probiotic yogurt
• Annex of the proposal (AGRI/38 743/2003rev3) by the European Commission for

a council regulation establish that ‘yogurt’ is a product obtained by the
fermentation of milk with cultures of Streptococcus thermophilus and
Lactobacillus delbrueckii sp. bulgaricus.
• The term ‘yogurt-like product’ is defined as alternative culture yoghurt (i.e. when
L. bulgaricus is substituted by other Lactobacillus species for the fermentation of
milk) or yoghurt containing probiotic bacteria (when probiotic bacteria are added
to the yoghurt or alternative cultures).
• ‘Probiotic bacteria’ are live food supplements, which benefit the health of the
consumer.
4
1. Probiotic yogurt (cont’d)
• The definitions from the previous proposal clearly differentiates between yogurt
starter cultures (S. thermophillus and L. bulgaricus) and probiotic bacteria that
may be added.
• But the distinction does not exclude S. thermophillus and L. bulgaricus from the category of
probiotic bacteria.
• The starter cultures can also be probiotics as long as they comply with the definition of
probiotic.
5
6
Yogurt production
• Yogurt is a fermented milk product that has been prepared traditionally by

allowing milk to sour at 40 – 45oC.
• Modern yogurt production is a well-controlled process that utilizes ingredients of
milk, milk powder, sugar, fruit, flavors, coloring, emulsifiers, stabilizers, and
specific pure cultures of lactic acid bacteria (S. thermophilus and L. bulgaricus) to
conduct the fermentation process.
7
8
Video (homemade Greek yoghurt)
https://www.youtube.com/watch?v=TfUtQuPfxxg
9
https://www.youtube.com/watch?v=1ZSoYrHyX9c
Yogurt production (cont’d)
• S. thermophilus and L. bulgaricus exhibit a synergestic relationship during the processing of

yogurt, with the ratio between the species changing constantly (Radke-Mitchell & Sandine, 1984).
• During fermentation, S. thermophilus grows quickly at first, utilizing essential amino acids
produced by L. bulgaricus.
• S. thermophilus, in return, produces lactic acid, which reduces the pH to an optimal level for
growth of L. bulgaricus.
• The lactic acid produced, and lesser amounts of formic acid stimulate the growth of L. bulgaricus.
The streptococci are inhibited at pH values of 4.2–4.4, whereas lactobacilli tolerate pH values in
the range of 3.5–3.8.
• After approximately 3 h of fermentation, the numbers of the two organisms should be equal.
With longer fermentation, the growth rate of S. thermophilus declines while L. bulgaricus
continues to reduce the pH by producing excessive amounts of lactic acid.
11
12
All yoghurt contains live bacterial cultures, which ferment milk to
make yoghurt, but only some types of yoghurt have added
probiotics (bacteria that have a proven health benefit).
Does all yoghurt

have probiotics in it?
Bio-yogurt
• Starter bacteria of conventional yoghurt (L.
bulgaricus and S. thermophiles) are not able to
subsist passing the intestinal tract and do not
colonize in the human gut.
• In recent years some yogurt products have been
reformulated to include live strains of L.
acidophilus and species of Bifidobacterium
(known as AB-cultures) in addition to the
conventional yogurt organisms, S. thermophilus
and L. bulgaricus.
• Therefore, bio-yogurt is yogurt that contains
live probiotic microorganisms, the presence of
which may give rise to claimed beneficial health
effects.
14
Bio-yogurt
• Bio-yogurt, containing L. acidophilus and B. bifidum (AB-yogurt), is a potential

vehicle by which consumers can take in probiotic cells.
• The number of probiotic bacteria required to produce a beneficial effect, has not
been established.
• Kurmann and Rasic (1991) suggested to achieve optimal potential therapeutic
effects, the number of probiotic organisms in a probiotic product should meet a
suggested minimum of >106 cfu mL-1.
15
Bio-yoghurt
• When should we add the probiotics into the yoghurt to retain the
viability of bacteria?
a. Before adding the starter
b. Together with the starter
c. After starter fermentation
d. Anytime
Simões Bandiera, Nataly, et al. Archivos latinoamericanos de Nutricion 63.1 (2013): 58-63.
Challenge in probiotics yoghurt
• When probiotic bacteria is added to yoghurt, it will have the impacts

on:
1. viability of probiotic bacteria during processing and the storage
period;
2. incubation time needed compared with conventional yoghurt;
3. organoleptic traits of the yoghurt
Meybodi, et al., International Dairy Journal 109 (2020): 104793.

Bio-yoghurt
• Factors affecting viability of probiotic bacteria in yoghurt:

1. Food components (macro- and micronutrients, flavoring agents, antioxidants,
amino acids)
2. Processing-related factors (pH, heat treatment, fermentation temperature,
packaging)
3. Microbiological factors (type of strains and inoculum level)
Meybodi, et al. International Dairy Journal 109 (2020): 104793.

Strategies to increase viability in bio-yoghurt
• There are some strategies to increase viability of probiotic bacteria in

yoghurt:
• Microencapsulation → good microencapsulation should be insoluble
in water, resistant to the pH of yoghurt and stomach, release the
living bacteria in the intestine, cost effective, size <100 μm,
acceptable by sensory.
• Prebiotic addition → adding fruits/ non-digestible carbohydrate into
probiotic yoghurt protects the probiotic bacteria by changing
buffering capacity

Health benefits of probiotic yoghurt
• Easier to be digested compared to milk due to autodigestion of

lactose by yoghurt bacteria improves its absorption in lactase-
deficient people
• Diarrhea prevention due to greater tolerance for pathogens (such as
Salmonella, Yersinia, and Helicobacter)
• Stimulation of immune system
• Prevention of non-communicable diseases, such as obesity, cancer,
type 2 dm, and cardiovascular diseases

Kaur, H., Kaur, G., & Ali, S. A. (2022). Fermentation, 8(9), 425.
Kaur, H., Kaur, G., & Ali, S. A. (2022). Fermentation, 8(9), 425.
2. Probiotic cheese
• Consumption of cheese has grown in the past decade in most countries,

unrelated to the socio-economic level of the country.
• A production of 17,778 million tons (t) in 2004 was reported, which corresponds
to a growth of almost 3272 t in the last decade.
• Cheese from cow’s milk represents 95 – 96% of the total cheese production
(International Dairy Federation – IDF, 2005)
24
2. Probiotic cheese (cont’d)
• In fact, cheese provides a valuable alternative to fermented milks and

yogurts as a food vehicle for probiotic delivery, due to certain
potential advantages.
• It creates a buffer against the high acidic environment in the
gastrointestinal tract, and thus creates a more favorable environment
for probiotic survival throughout the gastric transit, due to higher pH.
25
• Whey Portuguese cheese (requeijao) has been reported as a food vector for
environmental conditions prevailing in the gastrointestinal tract.
• Nevertheless, the results were strain-dependent, and loss of viability of different
strains (Bifidobacterium animalis strains e BLC-1, Bb-12 and Bo, Lactobacillus
acidophilus strains e LAC-1 and Ki, L. paracasei LCS-1, and Lactobacillus brevis
LMG6906) was observed after passage through an acidic hydrochloric (pH
2.5e3.0) and pepsin (1000 units/mL) solution at 37oC and a bile salts 0.3% (w/v)
solution (Madureira et al., 2005).
26
• The presence of the prebiotics inulin and oligofructose was described to

promote increased growth rates of bifidobacteria and lactobacilli, besides
increased lactate and short chain fatty acids production in petit-suisse cheese
supplemented with these microorganisms and submitted to batch culture
fermentation with human fecal slurry (Cardarelli, Saad, Gibson, & Vulevic, 2007).
27
Challenges in probiotic cheese processing
Some health benefits of probiotic cheese
• Probiotic fresh cheese (Argentinean fresh cheese) containing L. acidophilus A9, B.

bifidum A12 and L. paracasei A13 demonstrated immunomodulating capacity in
mice, providing increased phagocytic activity in the small intestine of peritoneal
macrophages, after 2, 5 and 7 days of its ingestion.
• Additionally, a significant increase in the number of IgA producing cells in the
large intestine after 5 days of administration was reported (Medici, Vinderola, &
Perdigon, 2004).
29
Some health benefits of probiotic cheese (cont’d)
• Hatakka et al. (2007) investigated the effect of consumption of probiotic cheese

on the oral candidosis for elderly people.
• Cheese containing a mixture of probiotic cultures (L. rhamnosus GG, L.
rhamnosus LC705, Propionibacterium freundenreichii spp., and Shermani JS) was
ingested by 92 elderly people.
• They suggested probiotic cheese could be used as a prophylactic approach for
decreasing the risk of hyposalivation and the feeling of dry mouth, and that it
could be considered beneficial to oral health in general.
30
3. Probiotic ice
cream
• Ice cream is a dairy product with good potential to act
as a food carrier for probiotic bacteria.
• Compared with fermented milks as a vehicle, ice cream
supports considerably greater viability of probiotic
strains during production and especially storage.
• Supplementation of ice cream with probiotic bacteria
has been found to have little effect on its flavor, texture
or other sensory characteristics.
Mohammadi, et al. Annals of microbiology 61 (2011): 411-424.

Pimentel, T. C., de Oliveira, L. I. G., de Souza, R. C., & Magnani, M. (2022International Journal of Dairy Technology, 75(1), 59-76.
In-class activity
• Determine the effect factors below on probiotics’ viability:

• Ice cream formulation
• Freezing process
• Overrun process
• Storage duration
• Ice cream melting during consumption
• What are the effects on viability?

• Best condition to retain viability?
Health benefits of probiotic ice cream
Pimentel, T. C., de Oliveira, L. I. G., de Souza, R. C., & Magnani, M. (2022International Journal of Dairy Technology, 75(1), 59-76.
Good to read
https://onlinelibrary.wiley.com/doi/epdf/10.1111/1471-0307.12821
4. Health benefits of probiotics, prebiotics, and synbiotics
(Gut-brain axis)
• The gut-brain axis (GBA)
consists of bidirectional
communication between the
central and the enteric nervous
system, linking emotional and
cognitive centers of the brain
with peripheral intestinal
functions.
• Neurons are cells found in your
brain and central nervous
system that tell your body how
to behave. There are
approximately 100 billion
neurons in the human brain
and 500 millions neurons in
gut, connected to your brain
through vagus nerve.
Have you ever felt
stomachache when
nervous?
Did you know probiotic
supplementation reduce
risk of stress?
Role of microbiota in gut-
brain axis
• Gut microbiota increases the production of
neurotransmitter (gamma-aminobutyric acid) in gut
and, hence, reduces risk of stress.
• Gut microbiota decreases the inflammatory markers,
such as cytokines, TNF-alpha, IL-1beta
• Prebiotics fermentation by gut microbiota increases
the production of GLP-1, PYY, CCK → appetite and
metabolism
Foster, J. A., Rinaman, L., & Cryan, J. F. (2017). Neurobiology of stress, 7, 124-136.
References
• Guarner, F., Perdigon, G., Corthier, G., Salminen, S., Koletzko, B., & Morelli, L.
(2005). Should yoghurt cultures be considered probiotic?. British Journal of
Nutrition, 93(06), 783-786.
• Putt, K.K., et al. (2017). Yogurt inhibits intestinal barrier dysfunction in Caco-2
cells by increasing tight junctions. Food and Function, 8: 406– 414.
43
Session 5
Carbohydrate-based
functional foods


1. Explain about carbohydrate-based functional foods
2. Explain about the effect of carbohydrate-based functional food to
human health
2
Course outline

1. Carbohydrate-based functional foods
a. Dietary fiber
b. Whole grain
c. Resistant maltodextrin
2. Carbohydrate-based functional foods health benefits
3
What comes into your
mind when you hear
“fiber”?
Definition of Dietary fibers
• Dietary fiber consists of carbohydrate polymers with ten or more monomeric units which are not
hydrolyzed by the endogenous enzymes in the small intestine of humans and belong to the
following categories:
• Edible carbohydrate polymers naturally occurring in the food as consumed;
• Carbohydrate polymers which have been obtained from food raw material by physical,
enzymatic or chemical means and which have been shown to have a physiological effect of
benefit to health as demonstrated by generally accepted scientific evidence to competent
authorities, and;
• Synthetic carbohydrate polymers which have been shown to have a physiological effect of
benefit to health as demonstrated by generally accepted scientific evidence to competent
authorities.
5
Dietary fiber recommendation
What is the fiber recommendation for Indonesian adults?

Fiber-rich foods
Source: IFT
7
Dietary fiber
classification
https://europepmc.org/article/pmc/4325053
Dietary fiber
According to its solubility in water:

• Insoluble dietary fiber ingredients → Celulose, hemicellulose, and
lignin.
• Cellulose is the main constituent of plant cell walls, and exists abundantly in
vegetables, fruits, cereals and legumes.
• Oat bran, chitin, etc.
• Soluble dietary fiber → Viscous (Pectin, β-glucan, Seaweed
polysaccharide, Microbial polysaccharide (xanthan gum)); Non-
viscous (inulin, resistant starch, polydextrose, FOS)
9
Dietary fiber
• Resistant starch → Resistant starches (RS) are a variety of starch that can resist digestion and
pass through the gastrointestinal tract
10
2. Carbohydrate-based functional foods
• Whole grain concept

• Fiber-rich foods
• Resistant maltodextrin
11
Whole wheat
• For the past century, most cereal products have been based on flour that consists
mainly of the endosperm, that is, after removal of the outer parts of the kernel,
bran and germ.
• Bran and germ are containing most of the dietary fiber and other bioactive
components such as micronutrients and phytochemicals. → Levels of these
substances in whole wheat flour are 2.5 to more than five times higher than
those in refined white flour.
12
Antioxidant Content of Whole Grain Breakfast Cereals,
Fruits and Vegetables
Miller et al., 2000
13
Whole grain concept
• A grain is considered to be a whole

grain as long as all three original
parts — the bran, germ, and
endosperm — are still present in
the same proportions as when the
grain was growing in the fields.
• Some countries have different
definition of whole grain with other
countries.
14
15
16
• Consumers worldwide are becoming increasingly interested in healthy eating, and have
consequently (re)discovered the value of wholegrain-based and fiber-rich products.
17
Consumer Preferences Towards Claims
From Food Marketing Institute, U.S. Grocery Shopper Trends, 2014

19
Which grain is “healthiest”?
• Each grain has its own little specialties:

• Oats are high in manganese (formation of bone)
• Durum wheat has more selenium (prevent cellular damage from free
radicals)
• Barley is highest in fiber
• Quinoa rich in folate
20
Table 1.1 shows how the launches of new wholegrain products have
grown exponentially over the past decade.
21
Health benefit of whole grain
24
Rave et al., 2007
25
26
A whole grain is a cereal grain that contains
the germ, endosperm, and bran, in contrast to
refined grains, which retain only the Tighe et al., 2010
endosperm.
27
Health benefits of whole grain (summary)
Călinoiu, L. F., & Vodnar, D. C. (2018). Nutrients, 10(11), 1615.

Sources of wholegrain
• Amaranth (Amaranthus spp.)
29
Wholegrain
• Barley (Hordum vulgare)
30
Wholegrain
• Buckwheat (Fagopyrum esculentum), etc.
31
Product
32
Can you guess??
#1. What are Whole Grains?
a. Grains that are darker in color.

b. Grains that are low carb.
c. Grains that contain the bran, germ and
endosperm.
d. Grains that are organic.
e. Grains that have been fortified.
#2. Which are always whole grain?
a. Barley
b. Oatmeal
c. Enriched Wheat Flour
d. 100% Wheat Bread
e. All of the above
f. None of the above
#3. Which guarantees a half serving or more of
Whole Grain?
a. d.
b.
c. e.
#4. True or False?
Whole grain products are always
high in dietary fiber.
a. True
b. False
Resistant maltodextrin (RMD)
• RMD is classified as type IV Resistant Starch, which is chemically

modified to alter its structure and digestibility.
• RMD is partially indigestible carbohydrate polymer due to the
presence of 1,2-, 1,3-, or 1,4- α and β-glycosidic linkage.
• RMD can be derived from potato, corn, wheat, and tapioca.
Evaluation of RMD health benefits on blood glucose
and satiety
Acute evaluation of resistant

maltodextrin
• Healthy subjects (n=16)
• Fasting blood glucose <100 mg/dl
07.30 08.00 11.00

a.m. a.m. a.m.
• • • • • • min
0 30 60 120 180
Anthropometry Test drink Blood drawing

Blood drawing consumption Subjective appetite
Subjective appetite
Stool form and gastrointestinal symptoms were evaluated 24 h following test day
Acute effect studies of RMD on subjective appetite and food intake
Satiety
Author Duration Subjects Dose Result
Emilien et al., Acute effect Healthy adults 0, 20, or 40 g No changes on appetite
(2018) (150 min) & food intake during
150 min
SCFA Ye et al., (2015) Acute effect Healthy adults 0,5, or 10 g RMD 10 g delayed
GLP-1
PYY
(240 min) hunger and increased
RMD satiety at 90 and 120
min.
Monsivais et al., Acute effect Healthy adults 12 g Only corn soluble fiber
Ye et al.,2015
(2011) (220 min) dextrin, but not RS,
polydextrose,
decreased appetite
Food intake at lunch
Effect of fermentable fiber
Developed nutritional supplement using RMD
Effect of maltodextrin replacement using RMD on physical properties of nutritional supplement
Original= 100% carbohydrate source from maltodextrin

RMD15= 15% carbohydrate source from RMD + 85% from maltodextrin
Developed nutritional supplement using RMD
Effect of maltodextrin replacement using RMD on physical properties of sensory acceptability
Original= 100% carbohydrate source from maltodextrin

Health benefits of resistant maltodextrin
Benefits Source of RMD Author (Year)

Glucose lowering effect Corn Hashizume et al., 2012
Wheat Nazare et al., 2011
Potato Wakabayashi et al., 1997
Lipid lowering effect Corn Hashizume et al., 2012
Wheat Li et al., 2010
Corn Kishimoto et al., 2007
Control appetite and food Corn Ye at al., 2015
intake Wheat Nazare et al., 2011
Wheat Guerin-Deremaux et al., 2011
Improve gut health Tapioca Sorndech et al., 2018*
Corn Burns et al., 2018
Corn Abellan Ruiz et al., 2015
Wheat Van den Heuvel et al., 2005
Improve mineral absorption Corn Miyazato et al., 2010*
Anti-inflammatory effect Corn Hashizume et al., 2012
References
• Miller, H. E., Rigelhof, F., Marquart, L., Prakash, A., & Kanter, M. (2000).
Antioxidant content of whole grain breakfast cereals, fruits and vegetables.
Journal of the American College of Nutrition, 19(sup3), 312S-319S.
54
Session 6
Lipid-based
functional foods


1. Explain the sources of lipid and lipid related functional foods
2. Explain about the health benefits of lipids and lipid related functional foods
3. Explain about meat-based functional foods
2
Course outline

1. Introduction
2. Health benefits of lipid
3. Dietary sources
4. Meat-based functional foods
3
1. Introduction
• Lipid can be defined as a fatty or waxy organic compound that, in

general, are soluble in ether, chloroform, or other organic solvents
but sparingly soluble in water.
• Lipids serve as structural components of biological membranes and
provide energy in the form of triacylglycerol (TAG) also called as
triglycerides.
4
1. Introduction (cont’d)
Major Component
R1, R2, R3 = identical or different fatty acids with even/odd numbers of carbon atoms
5
• The term lipids, fats, and oils are often use interchangeably.
• Fats generally refer to those lipids that are solid at room temperature and oils
generally refer to those lipids that are liquid at room temperature.
• The definition of lipids simply based on solubility is not justifiable and should be
widened to include compounds that are related closely to fatty acid derivatives
through biosynthetic pathways or by their biochemical or functional properties.
6
Classification of Fatty Acids
Fatty acids can be categorized based on:

• Degree of saturation
• Length of the carbon chain (short, medium, or long)
• Number of double bonds (unsaturated, mono,-or polyunsaturated)
• Essentiality in the diet (essential or non-essential)
7
Nomenclature of
Commonly Available n-3 PUFA
8
Line Drawing Representation of n-3 and n-6 PUFA Where Each
Angle Represents A Carbon Atom
9
• Lipids are considered in relation to excess energy balance, obesity, and as dietary
factor in the development of cardiovascular disease and many other harmful
disorders.
• However, not all lipids are bad and damaging to human health:
• PUFA
• Fat-soluble vitamins carrier
• Others?
• One food product can contain various types of lipids
10
Saturated and unsaturated fatty acids in the flesh of
L. vannamei
Gunalan et al. 2013

11
2. Health benefits of lipids
12
Results: effect on behavior
Results: effect on brain biochemical
parameters
15
16
18
Do we need to limit cholesterol intake below 300
mg per day?
a. Yes
b. No
Type of fat that should be limited?
a. Saturated fatty acid
b. Polyunsaturated fatty acid
c. Monounsaturated fatty acid
d. Trans fatty acid
What is predominant fatty acid in coconut oil?
d. Trans fatty acid
What is predominant fatty acid in palm kernel oil?
d. Trans fatty acid
level of triglycerides is significantly lowered in the body by consuming 3–4 g/d omega-3 fatty acids
Khan, M. I., Anjum, F. M., Sohaib, M., & Sameen, A. (2013). Reviews in Endocrine and Metabolic Disorders, 14, 287-297.
Khan, M. I., Anjum, F. M., Sohaib, M., & Sameen, A. (2013). Reviews in Endocrine and Metabolic Disorders, 14, 287-297.
DHA supplementation and fetal health
• EPA and DHA supplementation during pregnancy and breastfeeding may
protect their children against allergies.
• This may be due to the fact that fish-oil supplementation has been associated
with decreased levels of body cells associated with inflammation and immune
response
• In a study about food allergy and IgE-associated eczema, the period prevalence
of food allergy was lower in the maternal EPA+DHA supplementation group
compared to placebo (P < 0.05), and the incidence of IgE-associated eczema
was also lower in the maternal EPA+DHA supplementation group compared to
placebo (P < 0.05)
Swanson, D., Block, R., & Mousa, S. A. (2012). Advances in nutrition, 3(1), 1-7.
Journal reading
http://dx.doi.org/10.1016/j.jpeds.2016.02.077
28
Milk Fat Globule
29
3. Dietary Sources
30
EPA and DHA
content of
Malaysian
fish
Credit to: Pennington Biomedical Research Center (2005)
Omega-3 Fatty Acids
• Alpha-linolenic acid (ALA)

• Eicosapentaenoic acid (EPA)
• Docosahexaenoic acid (DHA)
Credit to: Pennington Biomedical Research Center/PBRC (2005)
Alpha-linolenic acid (ALA)
• Scientific abbreviation is 18:3n-3

• The first part (18:3) suggests that ALA is an 18-carbon fatty acid with 3 double
bonds
• The second part (n-3) tells you that ALA is an omega-3 fatty acid
• It is required for health, but cannot be synthesized in humans
• Must be obtained from the diet
34
Alpha-linolenic Acid
• Humans can synthesize other omega-3 fatty acids from ALA:

• Eicosapentaenoic acid (EPA): 20:5n-3
• Docosahexaenoic acid (DHA): 22:6n-3
• These two are usually referred to as marine-derived omega-3 fatty

acids because they are abundant in certain species of fish
• Whereas, ALA is considered a plant-derived omega-3 fatty acid

FOOD SERVING ALPHA-LINOLENIC ACID (G)
Flaxseed oil 1 tablespoon 8.5
Walnuts, English 1 ounce 2.6
Flaxseeds 1 tablespoon 2.2

Sources of
Walnut Oil 1 tablespoon 1.4
Alpha-
Canola Oil 1 tablespoon 1.2
linolenic acid Mustard Oil 1 tablespoon 0.8
Soybean Oil 1 tablespoon 0.9
Walnuts, Black 1 ounce 0.6
Olive Oil 1 tablespoon 0.1
Broccoli, raw 1 cup, chopped 0.1

Sources of EPA and DHA
Food Serving EPA (g) DHA (g) Amt providing 1
g of EPA + DHA
Herring, Pacific, cooked 3 ounces 1.06 .75 1.5 ounces
Salmon, chinook, cooked 3 ounces .86 .62 2 ounces
Salmon, Atlantic, cooked 3 ounces .28 .95 2.5 ounces
Oysters, Pacific, cooked 3 ounces .75 .43 2.5 ounces
Salmon, sockeye, cooked 3 ounces .45 .60 3 ounces
Trout, rainbow, cooked 3 ounces .40 .44 3.5 ounces
Tuna, white, packed in 3 ounces .20 .54 4 ounces
water
Crab, dungeness, cooked 3 ounces .24 .10 9 ounces
Shrimp, cooked 3 ounces .15 .12 11 ounces
Cod, Pacific, cooked 3 ounces .09 .15 12.5 ounces
Fish oil, menhaden 1 gram .13 .09 5 grams
Fish oil, salmon 1 gram .13 .18 3 grams
Kaur, N., Chugh, V., & Gupta, A. K. (2014Journal of food science and technology, 51, 2289-2303.
Docosahexaenoic acid (DHA)
• Found in very high concentrations in the cell
membranes of the retina
• It conserves and recycles DHA even when

omega-3 fatty acid intake is low
• Studies in animals indicate that DHA is

required for the normal development and
function of the retina
3. Dietary source: Olive oil
• Olive oil is extracted from fresh or ripe fruits of the long lived ever green olive
tree Olea europaea, which originated in the Mediterranean area.
• The oil is regarded as a healthy dietary oil because of its high content of
monounsaturated fat (C18:1).
• It is produced principally in Greece, Italy, Spain, France, Turkey, Portugal, Tunisia,
Morocco, and USA (California).
• These countries have lower incidence of coronary heart disease (CHD), and is
postulated due to their Mediterranean diet (including large amounts of olive oil).
40
Olive oil
• Main consuming countries are also the main olive oil producers. →
EU accounts for 71% of world consumption.
• Olive oil contains 47% water 31% solids and 22% oil.
41
42
Olive oil extraction
43
Video – olive oil extraction
https://www.youtube.com/watch?v=wvacR3rTgFU
44
4. Meat-based functional foods
• Meat is often associated as one type of unhealthy food due to the

relationship between its constituents with some diseases
• However, nutrients like iron, vitamin B12 and folic acid have inferior
bioavailability in food other than meat
• Current need: healthier meat products
• Animal processing
• Food product processing: raw materials, variation and designing new
formulations of meat products which have improved nutritional profile
45
DOI 10.1002/mnfr.200700367
46
Meat-based functional food:
omega-6/omega-3
• Ratio omega-6/omega-3
• Conventional meat product: 15
• Healthy effect ~ <4
• Strategy: addition of omega-3 PUFA
• Objective: to design new functional meat products
based on a healthy combination of omega-3 PUFA and
supercritical rosemary extracts with antioxidant
activity (AA)
47
Meat Products + Treatments
Most of fat Deodorized Salmon Oil

was
removed
Frankfurt sausage, ham, cooked meat

Supercritical rosemary extract
48
49
Lipid oxidation measurement:
Thiobarbituric acid reactive substances (TBARS) value
50
Let’s observe and (critically) think!
• Do you know any similar example(s) of Indonesian

meat product?
• Can it be categorized as “meat-based functional
food”? In which way?
• Improvement? Research and Development?
51
References
• Chapter 5 – Guo, M. (2009). Functional foods: principles and technology. CRC

press.
• Guston, L.D. (2012). Lipids for functional foods and nutraceuticals. Woodhead
Publishing.
• Related scientific journals.
52
Session 7
Marine-based
compounds as
functional food
ingredient

1. Explain the sources of marine-based bioactive compounds, and
their potential uses
2. Explain the role of biotechnology in the development of functional
foods
2
Outline
1. Introduction of marine bioactive compounds

2. Trends in marine bioactive compounds
3. List of marine-based bioactive compounds, its sources and potential
uses
4. Role of biotechnology in the development of functional foods
3
1. Introduction of Marine
Bioactive Compounds
4
Introduction of Marine Bioactive Compounds
Definition
• Compounds that have effects on a living organism,
tissue, or cell
• Completely distinguished from essential nutrients
Characteristic
• Commonly sold, consumed, and marketed in form of
raw ingredients, foods, and even supplements
• Daily consumption of bioactive compounds may
provide additional health benefit.
5
Spinal Muscular Atrophy (SMA)
6
7
Terra et al., 2014
2. Trends in Bioactive
Compounds
8
Trends in Healthy Foods in relation to Bioactive
Compounds
9
10
Statistical Analysis of New Marine Natural Products
from 1985 - 2012
11
Statistics of bioactivity of new marine natural products by year
(* PHVD: Prevention of heart and vascular disease, ** PN/NT:
Protection of neurons/neurotoxicity).
12
[a]
Distribution of multiple bioactive compounds

in each category of chemical compounds
13
[b]
Analysis of the bioactive compounds according to their chemical

structures and the proportion of different activities in each type
of compound (* PHVD: Prevention of heart and vascular
disease, ** PN/NT: Protection of neurons/neurotoxicity).
14
Summary from The Statistical Data
Anti-cancer
Anti-bacteria
Anti-fungus
Anti-virus
Pest resistance
PHVD (Prevention of Heart and Vascular Disease)
PN/PT (Protection of Neuron / Neurotoxicity)
Other Activities
15
3. List of marine-based
bioactive compounds, its
sources and potential uses
16
Freitas, A. C., Rodrigues, D., Rocha-Santos, T. A., Gomes, A. M., & Duarte, A. C. (2012). Biotechnology advances, 30(6), 1506-1515.
Algae
- anti obesity effect -
18
https://doi.org/10.3390/nu12123798
19
L-Fucose suppresses fat accumulation in high-calorie diet
(HCD)-induced obese mice
(A) Body
weight gains in high-calorie diet-induced
obese 6 mice fed either a control high-
calorie or high calorie diet with different
concentrations of L-fucose
(B) Visceral fat weight
(C) Liver fat mass of mice detected by CT-

scan
20
L-Fucose decreases the expression of adipogenic genes
Relative gene expression in (A) liver, (B) skeletal muscle, (C) adipose tissue of
glucose and L-fucose-treated mice
21
22
Dried Bonito (Katsuobushi)
-Antihypertension-
23
Katsuobushi or Bonito
Flakes → simmered,
smoked and fermented
skipjack tuna
24
Effects of Dried Bonito (Katsuobushi) and Captopril on Rat Isolated Aorta
25
Effects of Dried Bonito Extracts and Captopril on Angiotension
I-Induced Contractions in the Rat Isolated Aorta
https://doi.org/10.1271/bbb.69.911
26
Small Dried Sardine Extract
- anti-allergic effect -
30
Awane et al. Unpublished data
Effect of ESDS extract on degranulation and cell viability

of RBL-2H3 cells
・ Degranulation assay
40 *p<0.05 vs. control
β-Hexosaminidase release (%) 35
30
25 *
20 *
Degranulation is a cellular
15
process that releases
10 antimicrobial cytotoxic or
5 other molecules from
secretory vesicles called
0
Control 180 360 720 1440 granules found inside
Protein conc. (µg/mL) some cells.
ESDS extract suppressed antigen-induced

degranulation without cytotoxicity.
31
Awane et al. Unpublished data
Effect of ESDS extract on cedar pollinosis model mice
Rubbing frequency Sneezing frequency

175 *p<0.05, vs. control
30
Rubbing frequency
150
(times /30 min)
Sneezing frequency
25
(times /30 min)

125
20
100
15 *
85
10
50
25 5
0 0
Intact Control Sample Intact Control Sample
ESDS extract relieved allergic symptoms caused by

cedar pollinosis.
32
34
35
Hamed et al., 2015
36
37
Potential Use
Functional Foods
• Bioactive peptides
• Bioactive polysaccharide
• Prebiotics
Supplements
• Essential Fatty Acids (EPA & DHA)
• Vitamins & Minerals
Drugs
• Anti-cancer
• Antibiotic (bacteria/fungi/virus)
• Anti-inflammatory
38
eicosapentaenoic acid (EPA)
39
• Kao Corporation (President and CEO: Michitaka Sawada)
launched Healthya Coffee, the first coffee beverage from
the Healthya series, on April 4.
• The new drink contains a high level of polyphenol "coffee
chlorogenic acid" and is approved as Food for Specified
Health Use (FOSHU) due to its effect enhancing the body
fat utilization as energy.
Fortified drinks for
more health benefits?
40
Conclusions
Main Bioactive Compounds
• Polysaccharide
• Peptides
• Essential Fatty Acids
• Other compounds (pigments, phenolic groups, vitamins & minerals, sterol / stanol)
Sources (Marine)
• Micro-/Macro-algae
• Invertebrates and Fishes
• Microorganism (bacteria)
Functions
• Anti-cancer
• Anti-inflammatory and Prevention of Disease
• Anti-pathogen (bacteria, fungus, virus)
Potential and Marketability
• Utilization of marine bioactive compounds is already flexible, as long as it is supported by robust scientific
evidences
• Increasing trends in terms of functional foods, supplements, and therapeutic drugs, corresponding to healthy
lifestyle and evolving diseases
41
Suggestion / Recommendation
• Tools for “Data-mining/mapping/processing” for secondary existing data

from various scientific sources.
• Reliable and feasible extraction/isolation methods for obtaining targeted
bioactive compounds → culture dependent / independent
• Mass-production of these bioactive compounds has to be as sustainable as
possible, meaning that how to maximally forage marine sources without
damaging the aqua-environment
• Marine proteins / peptides may cause allergies to certain group of people,
claim and labeling of marine-derived functional foods have to be proper.
• For drugs, clinical trials is a must!
42
4. Role of biotechnology in the development of
functional foods
43
functional foods
44
functional foods
Journals reading:
• Golden Rice is an effective source of vitamin A
• Marine biotechnology advances towards applications in new
functional foods
45
References
1. Barde et al., (2015). Marine bioactive agents: a short review on new marine antidiabetic
compounds. Asian Pacific Journal of Tropical Disease. Volume 5, Supplement 1 Pages S209-S213
2. Suleria et al., (2016). Marine bioactive compounds and health promoting perspectives;
innovation pathways for drug discovery. Trends in Food Science & Technology Volume 50 Pages
44–55
3. Rocha-Martin et al., (2014). Emerging Strategies and Integrated Systems Microbiology
Technologies for Biodiscovery of Marine Bioactive Compounds. Mar. Drugs, 12, 3516-3559
4. Mehhub, et al., (2014). Marine Sponge Derived Natural Products between 2001 and 2010: Trends
and Opportunities for Discovery of Bioactives. Mar. Drugs, 12, 4539-4577
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Session 1

CLO1: Describe the definition, history, and global aspects of functional

• Late 10-15 min tap-in allowed, seat in allowed

• Purpose: writing practice and coaching

Group Topic Members

Group Topic Members

Group Topic Members

• Writing organization (15%)

• Purpose: boost creativity and apply your knowledge

International Submitted to the journal/ conference/ competition 80

Score Range Grade Status

By the end of this session, students will be able to :

Subtopics that will be delivered in this session:

“Let the food be thy medicine and

PALATABILITY NUTRITIOUS NO DOSAGE

The utilization of food for preventing nutrient-deficiency diseases was

• Functional foods are similar in appearance to conventional foods; the former

• The term "nutraceutical" was coined from "nutrition" and "pharmaceutical" in

• Nutraceuticals differ from dietary supplements in the following aspects:

Purwaningsih et al. 2021

*In older adults

*In older adults

Vicentini et al., 2016 51

• Oxidation is one of the metabolic reactions in the body and in foodstuffs

• Free radicals are atoms or molecules having unpaired

1. Normal aerobic respiration

vit E can be regenerated by vit C

vit A can also donate its electron and after that it 64

1. Accuracy in Accuracy in defining the terminology of functional food

3. Accuracy in describing biological antioxidants defense system

• Chapter 1 and 2 Guo, M. (2009). Functional foods: principles and technology.

By the end of this session, students will be able to :

Subtopics that will be delivered in this session:

• Hence, antioxidants are defined as

• The term “antioxidant” has many definitions

Huang, et al, J. Agric. Food Chem. 2005, 53: 1841-1856

• The term oxidizable substrate encompasses almost everything (except water)

• Antioxidants have become an indispensable group of food additives. The use of

Antioxidants in food are important for four reasons:

1. Primary or chain breaking antioxidants (scavenger antioxidants):

Ilavenil et al., 2011.

2. Secondary or preventive antioxidants. These antioxidants act

3. Tertiary antioxidants: These antioxidants remove damaged

• Small berries constitute an important source of potential health promoting

• The fruits of American cranberries, Vaccinium macrocarpon and European

• Proanthocyanidins and anthocyanins make up the pigment of the leaves, and

• Cranberry fruits serve as an excellent source of anthocyanins, flavonol glycosides,

Resveratrol inhibits PDEs (phospho

Downstream of AMPK, an increase in

• Urinary tract infections (UTIs)

• Recently, a study was completed in which extracts of purified cranberry

• Various bacteria appear to be major causative factors in the etiology of both

• In vitro and in vivo animal studies have found anti-inflammatory, anticarcinogenic,

• Grapes belonging to the species Vitis vinifera L. are predominantly cultivated in

• The tomato (Lycopersicon esculentum) is one of the world’s major vegetables

• Lycopene and β-carotene have been shown to act as powerful antioxidants in

• Bring to campus: 1 product that you think contains

By the end of this session, students will be able to :

Subtopics that will be delivered in this session:

• There is a long history of health claims concerning living microorganisms in food,

• Since 7000 BC, archaelogists reported

• FAO and WHO set the

An ideal prebiotic should be: