LTNIT 11 FOOD COMPONENTS OTHER THAN

ESSENTIAL NUTRIENTS
Structure
11.1 Introduction
11.2 Functional Foods
11.2.1 Classification
11.3 Bioactive Substances from Protein Foods
11.4 Non-Glycerides in Edible Oils
11.5 Probiotics and Prebiotics
11.5.1 Definition and Characteristics
11.5.2 Probiotics: Dietary Sources and their Mode of Action/Effects
11.5.3 Prebiotics: Dietary Sources and their Mode of Action/Health Effects
11.6 Polyphenols
11.6.1 Definition and Classification
11.6.2 Bioavailability of Polyphenols
11.6.3 Influence of Polyphenols on Macronutriellts and Minerals
11.6.4 Health Benefits of Polyphenols
11.7 Phytoestrogens
11.7.1 Dietary Sources and Chemical Forms
11.7.2 Physiological El'fects
11.8 Other Dietary Factors with Antinutritional Effects
11.8.1 Protease Inhibitors
11.8.2 Saponins
11.8.3 Amylase Inhibitors
11.8.4 Lectins or Haemagglutinins
11.8.5 Phylates
11.9 Health Benefits of other Dietary Factors with Antinutritional Effects
11.10 Let Us Sum UP
11.11 Glossary
11.12 Answers to Check Your Progress Exercises
--

11.1 INTRODUCTION
In the previous units so far, we have read about the six major nutrients, which are
essential for us. Though these vary in their requirements and roles, they have
significance in our daily diets. Apart from these nutrients, there are certain health-
promoting essential nutritional factors which have protective and preventive functions.
In this unit, we will be studying about what are these and their beneficial, as well as,
adverse health effects.

Theidea that food can be health-promoting beyond nutritional value is gaining a wide
acceptance. This broad view of nutrition has led to the concept of 'functionality'. The
American Dietetic Association (ADA) has taken the position that specific substance
in food e.g. phytochemicals may benefit health when consumed as a part of a varied
diet. Many common foods contain non-nutritive components that may provide protection
against chronic diseases. It is believed that 'nutraceuticals7 or 'functional foods'
will help to provide an increased 'health span', that is, medical benefits, including the
prevention and treatment of disease.
These substances are bioactive (extra-nutritional) constituents of foods which can act
as 'chemopreventers7 i.e. have anticarcinogenic and other beneficial properties evoking
 physiological, behavioural and immunologic effects. This unit deals with such health Food Components other
promotive nutritional factors and bioactive constituents- their potential health Nutrients
implications and mechanisms of action to the extent that has been elucidated so far.
While reading this unit you should bear in mind that despite animal research and some
clinical trials, there is still uncertainty about the absolute safety and long-term benefits
of supplementing the diet with some of these constituents. While functioilal foods in
general have pronloting properties, some of them may have an adverse effect on
absorption or utilizatioil of certain nutrients. We shall lcarn about these effects in this
unit.
j
Objectives
After going through this unit, you will be able to:
describe what are functional foods/nutraceuticals,

I
classify the functional foods,
0 discuss the potential health implications and mechanisms of action of functional
foods, and
explain the various adverse effects of these substances.

11.2 FUNCTIONAL FOODS

The term 'functional food' was born in Japan. Functional foods are actually products
formulated with naturally occuning cheiriicals or a combination of these. They are
found in many fruits, vegetables, grains, herbs and spices to provide a health benefit,
lower the risk of certain diseases or affect a particular body process. To be precise,
these are the food substances, beyond basic nutrients that are designed to lower
tlze risk or delay the onset of certain diseases. The Japanese were the first to
observe that food could have a role beyond nutrient supply. Thus, a functional food
must be a food and not a drug. Beneficial effects should be obtained by coilsulning
normal amounts that is within the parameters of a 'normal' diet.

In Japan, several functional foods are available in the market. It was the first country
to legislate these products (FOSHU stam1,-Foods of S'ecijied Health Use). Europe
and the American countries incorporated later the concept of an added value of food.
There is no consensus bet6een Europe and the USA regarding a concrete definition.
Therefore, we have several different terms such as: nutracsutical, designer food,
vita foods, pharma food nzedicinal foods, prescriptive foods, tlzerapeutic foods,
super foods, foodiceuticals ~rndmedijootls. Nutraceutical is the preferred Len11 in
USA. European experts decided to adopt the term 'functional food' with a consensus
definition. You may refer Lo 13ox 11.1 and Box 11.2 for a detailed European and
Japanese perspective on functional foods.

Box 11.1 Functional Food in Europe: The Functional Food Science in

Europe (PUFOSE, ILSI Europe, 1999)
Functional foods are:
conventionaYeveryday foods consumed as a part of the normal diet;
composed of natusally-occurring components, sometimes in increased
concentration or present in foods that would not normally supply them;
scientifically demonstratid to promote positive effects on target functions
beyond basic nutrition;
thought to provide enhancement of the state of well-being and health in order
to improve the quality of life andlor reduce the risk of disease; and
advertised by authorized claims.
Advance Nutrition
Box 11.2 Japanese 'FOSHU' Criteria for Functional Food

The Japanese criteria for functional foods include:

They are food (not capsules, pillslpowder) on the basis of naturally occurring
food components.
They can and should be consumed as part of noimal daily diet.
o They have a defined function on the human organism to:
- improve imnune function,
- prevent specific disease,
- support recovery from specific diseases,
- control physical and psychic comnplaints, and
- slow down the ageing process.

According to Japanese criteria, functional foods are not capsules, pillslpowder,

however, in some countries functional foods (prebiotic products) are marketed in the
fonn of powder or liquid suspension. Now then having gone through the European-
and Japanese perspective on functional food, it must be coining to your mind that
what is present in a food that makes it to be classified as a functional food. Let us
find out.

So far, the most important components that have been identified and can be added
to food are:
- Probiotics: A mono or mixed culture of living organisms, which when ingested
in certain amounts, has a positive impact on host health, beyond conventional
nutritional effects. You may recall reading about them earlier in the Food
Microbiology and Safety Course (MFN-003) as well in Unit 1. These stimulate
the growth of certain other bacteria in the colon, thereby improving health.
Bacteria most often used as probiotics are Lactobacilli and Bifidobnctel-in
which can be given along with the fermented foods e.g. yoghurt, fermented
vegetableslmeat.
- Prebiotics: Ingredients/compounds that have a beneficial effect on microflora
in the large intestine of the host e.g. fibre, fn~ctooligosaccharides, lactulose,
sugar alcohols. Generally, they are carbohydrates that may be fermented in the
large bowel and stimulate growth of potentially beneficial bifidobacteria.
Several beneficial effects of functional foods have been reported, which include
effects such as antioxidant, anticarcinogenic, blood glucosellipid-lowering, regulation
of intestinal transit, prebiotic effect, anti-bacterial, anti-viral and immunoyotentiating.
A reduced risk of several chronic diseases such as CVD, cancer, diabetes, hypertension,
osteoporosis has also been reported.

A wide variety of functional foods are available in the market. Functional foods can
be identifiedtselected on the basis of their properties, clinical significance or composition.
Discussed below are some of the common methods employed for classifying functional
foods.

11.2.1 Classification
Functional foods may be classified in various ways. From the nutritional viewpoint,
they can be categorized as nutrients and non-nutrients. Have a look at the Table
11.1, which presents this classification.
 Table 11.1: Classification of functional foods Food Components other
than Essential Nutrients
Nutrients Non-Nutrients
Lipids Fibre
n-3 fatty acids Insoluble and soluble fibre
Conjugated linoleic acid 0 Phenolic compounds
Vitanzins Phenolic acids
Folates Flavonoids
E Isoflavones
I Carotenoids: p -carotene and a-carotene ( Cntechins
Vitamin C Tannins
Minerals Non-digestible Oligosaccharides (NDO)
Selenium
I Fructails
Galacto oligosaccharides
Isomalto oligosaccharides
Xylo oligosilccharides
I I Soy oligosaccharides

Organ.osulpIz~ircompounds

Here we will not be discussing nutrients classified as functional foods, since these
have already been dealt with in the earlier units of this course. We will only focus
on the non-nutrient category. But first w e shall look at the other classifications of
functional foods.
Another classification is by the target orga~dsystenzbenefited. This is of clinical
relevance especially with regards to treatment and management of various diseases.
Examples of functional foods, on the basis of their beneficial impact on different
organ systems, are given in Table 11.2.
Table 11.2: Classitication based on orgadorgan system

Orgadorgan System Food Component

Gastrointestinal tract Prebiotics (NDOs), soluble fibre
Insoluble fibre
Probiotics
Polyphenols
Phytatc
n-3 fatty acids
Micronutrients
Cardiovascular system n-3 fatty acids
Polypheilols
Micronutrients
Soluble fibre
Immune system Prebiotics
Probiotics
Nutrients
n-3 fatty acids
Polyphenols
Skeletal system Fructans
Kidney Fructans
Advance Nutrition Besides classification based on orgadorgan systems, another classification is based
on origin or source i.e. plant, animal or microbial. Some compounds/substances
included in each group are listed herein Table 11.3.
Table 11.3: Classification based on origidsource
Source Components
hmal Conjugated linolenic acid (dairy products)
Chitosan
Fish oils (oL-fatty acids)
Microbial Probiotics
Plant Fibres (all components)
Polyphenols
Fructans
n-3 fatty acids
Phytates
Carotenoids
Non-glycerides in edible oils

Some foods are inherently functional and do not require much modification whereas
others do. Foods may be made functional by:
elimination of components e.g. toxins or allergenic proteins,
increasing the concentration of a natural component e.g. fortification,
addition of components with beneficial effects e.g. non-vitamin antioxidants,
addition of beneficial microbes e.g. some yeasts, bacteria,
replacement of a component, usually a macronutrient e.g. fat replaced with
modified or emulsified carbohydrates, and
enhancement of bioavailability of components.
Thus, a number of substances or components can be added to food to make it more
appropriate for the treatment of a disease or management of health. Next, we move
on to the study of functional foods. We will begin with the bioactive substances
especially those present in protein-rich foods.

11.3 BIOACTIVE SLTBSTANCES FROM PROTEIN

FOODS
Bioactive substances are derived from living organisms that can be used by humans
for a variety of applications. They are constituents in foods or dietary supplements,
'other than those needed to meet basic human nutritional needs and are responsible
for changes in health status. Let us in this section learn About these as present in
protein foods and their possible health effects.
The main function of dietary protein, as we know, is to supply the body with essential
amino acids and organic nitrogen. However, proteins also supply bioactive peptides
that are released by proteolysis in vivo (within the living body) and in vitro (in an
artificial environment outside the living). Casein-derived peptides have found applications
as dietary supplements and pharmaceutical preparations.
Peptides originating from food proteins should be considered as potential modulators
of various regulatory processes in the body. A number of in vitro studies have shown
anti-carcinogenic and anti-tumor activity for many of these in addition to the
antimicrobial activity of the whey proteins.
Let us next get to know more about edible oils and fats and how they act as bioactive
components.
 Food Components other
11.4 NON-GLYCERIDES IN EDIBLE OILS than Essential Nutrients

You may be aware of the fact that the glyceride fraction (an ester of glycerol and
fatty acids that occurs naturally as fats and fatty oils) in edible oils and fats is about
90-98% and the remaining 2-10% is the non-glyceride fraction and that it contains
a variety of components. These are: Sterols, terpene alcohols, tocopherols,
Izydrocarbons, long clzain alcohols including waxes, carotenoid pigments, suljur-
and nitrogen-containirzg,flavour compounds. They possess nutritiona1 and physiologic
functions. These conlponents are present in the crude extract and are destroyed/
reduced duiing the process of concentration or chemically modified during refining.
A biief review on these non-glyceride fractions follows:
e Sterols: These constitule a major ~jroportion of the non-glyceride component
while tocopherols, carotene pigments and flavour compounds are minor
compounds.
e Aliphatic alcohols: These are water-insoluble, will1 their content varying from
0.5-7% of the unsaponifiabIe matter. Rice bran oil contains a mixture of ferulic
acid, esters of slerols and triterpeile alcohols, 'Oryzanol' which is considered to
have a hypocholesterole~~~ic effect.
e Terpene alcohols: Partly free, partly esterified e.g. fer~~lic
acid; these are found
in significant amounts in rice bran oil, wheal germ oil, soybean oil, linseed and
olive oil. Commonly used edible oils e.g. groundnut, sesame, safflower, corn,
coconut, sunflower and palill oils contain only -10-20% of the amount found in
rice bran oil. A hypocholesterolernic effect has been seen in animals and humans.
Blends of rice bran oil and safflower oil have been shown to reduce cholesterol
and LDL aside from the effects of PUFA and MUFA.
So far we have read about functional foods, bioactive substances present in protein
foods and the non-glyceride fractio~isin edible oils which have been found to have
health promotive properties. We shall learn about some other food components in our
subsequent discussions. However, before we proceed, it would be a good exercise
to recapitulate about the concepts learnt so far by answering the questions mentioned
in the check your progress exercise 1.

Check Your Progress Exercise 1

1) What are functional foods? Where are these found?
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
2) What are the functions of functional foods, as per the Japanese 'FOSI-IU'
criteria?
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
Advance Nutrition
3) List the important components that .can be added to food.
.....................................................................................................................
.....................................................................................................................
4) Enumerate:
a) Any five 'non-nutrient functional foods'.
.................................................................................................................
.................................................................................................................
b) The food components for benefiting the iminune system.

.................................................................................................................
C) Any three plant-based functional foods.

.................................................................................................................
5) What are the different ways by which foods can be made functional?
.....................................................................................................................

.....................................................................................................................
6) What 'are the major components of non-glyceride fraction in edible oils and fats.
.....................................................................................................................
I
Let us now move to the next section on probiotics and prebiotics. We have earlier
briefly discussed about these. Let us now get to know about these in detail.

11.5 PROBIOTICS AND PREBZOTICS

We all have read or heard about probiotics and prebiotics at some point of time.
However, what exactly do we mean by these terms? When were they identified?
What are the benefits of these components and which foodslfood products would
contain them? These are a few questions which shall be answered through the
discussions to be followed next.
Well, during the initial stages, research and development of functional foods was
confined to mostly those to which certain components such as micronutrients were
added. However, one of the most promising current targets for functional food
development is the gastrointestihaltract (GIT). The human large intestine is inhabited
 by very large numbers of microorganisms. The resident microflora through their Food Components other
than Essential Nutrients
metabolic activities exerts important physiological action relevant to health and disease.
This and other possibly beneficial effects to other target organs have led to an
intensive study of probiotics and prebiotics.
We shall now begin our study about pro and prebiotics by first defining these terms.

11.5.1 Definition and Characteristics

An early definition of probiotics is 'organisms and substances which contribute to
intestinal microbial balance'. Later the word 'substances' was removed since these
could include antibiotics and microbial stimulants that are categorized as prebiotics.
The revised definition is a live micl-obinl feed supplement which beneficially
affects the host animal by improvilzy its intestinal microbial balance. It has been
suggested that the definition should be expanded to include other areas of the body
e.g. skin, vagina and respiratory tract. Thus, the beneficial effect of probiotics can
be mediated through the gut microflora by ingesting viable micsoorganisms. According
to the definition, probiotics include not only pseparatioils but also traditional yoghurts
and other fermented foods.

Prebiotics, on the other hand, are non-digestible ,food ingredients that be~zeficially
aflect the host by selectively stinzulating the growtlz and407 activif)~of one or a
limited nuinber o f bacteria in the colon.
Let us now move on to the understanding of the characteristics of probiotics and
prebiotics.
Characteristics of Probiotics and Prebiotics
The characteristics of probiotics and prebiotics are highlighted in Table 11.4.
Table 11.4 : Characteristics of probiotics and prebiotics
Probiotics Prebiotics
Ase rnicroorga~~isms Are non-digestible dietary/food
ingredients
Are not hydrolyzed/absorbed in the
upper GIT, reach the large intestine in
nn intact form and have a specific
metabolism therein - directed towards
beneficial /advantageous bacteria than
those which are advcrseJdetrimental for
host I~ealth
Promote a healthy intestinal microflora Fermented in colon
Ensure colonization resistance to pathogens Alter composition of colonic microbiota
towards a healthier community
Destruction of genoioxins and mutagens May induce systemic effects beneficial
Immune system function to host

Next, we shall s h ~ d yabout the mode of action, usage in foods and resistance properties
of probiotics.

1,
/
11.5.2 Probiotics: Dietary Sources and their Mode of Action/Effects
We can have specific preparations designed for probiotic use, as well as, foods.
Probiotic products which are marketed are in the form of powders, tablets and liquid
suspensions. Among the various organisms, lactic acid bacteria, streptococci and
bifidobacteri are commonly used.
Advance Nutrition It is not clear how probiotics influence the flora and produce a beneficial effect.
However, colonization of the gut appears to be a prerequisite for the probiotic effect.
Important relevant features may be:
- resistance to low pH and bile acids, and
- ability to adhere to intestinal epithelium.

However, ability to adhere to intestinal epithelium does not ensure that an organism
will permanently colonize the gut. Also, there is a continuous interchange of species
with one strain being replaced by another, which is possibly better suited. Since some
strains are more suitable than others. Organisms such as Saccharomyces boulardii
are effective although they do not grow in the gut. For such organisms, continuous
administration is required to ensure the presence of large numbers of metabolizing
cells in the GIT.

As a dietitian, you must be now curious to know which foods contain probiotics. Let
us read further and find out.

Probiotics in Foods I

Yoghurts have been supplemented with probiotic strains of bifidobacteria and

Iactobacilli. Milks fermenled solely by intestinal isolates of lactic acid bacteria have 1
also been developed. Other fermented milk products like cottage cheese, sour cream I
etc. contain viable organisms and may have an incidental probiotic effect.
I

We will now examine the health effects of probiotics.

Health ESfects related to Probiotics:
Several lines of evidence support the conclusion that normal gut microflora
are involved in resistance to disease, especially gastrointestinal infections. These
are:
- germ-free animals are more susceptible to infection than animals having complete
I
gut flora,
- orally administered antibiotics increase susceptibility of animals to infection
(antibiotics are known to destroy gut microflora), and
- administration of enemas of faecal suspensions from a healthy adult can control
antibiotic-associated diarrhoea (AAD). -,

Let us briefly examine the effects that have been reported (remember most of it has
been on animals):
I
I
!
* 0 In animals, probiotics may control tumour production.
In humans, probiotics can reduce incidence of intestinal infections. The possible
mechanism of action is: the probiotic has some antagonistic effect (directfindirect)
on the pathogen: either through direct chemical antagonism, competition for I

nutrients or an indirect effect via the immune system by competition for receplors
on the epithelial surface.
50% reduction in AAD observed in patients given Saccharomyces boulardii.
Significant reduction in diarrhoea incidence among children aged 4-45 months
after administration of L.casei GG. The effect was more pronounced in patients
with confirmed rotavirus infection.

1
o Can have an antifungal effect. During chemotherapy for leukemia. Treatment Food Components other
with a milk preparation containing L. ncidophilus and Bifiilobncteriunz sp. than Essential Nutrients
makedly reduced the faecal count of Candida.

Now that we have understood the benefits and usage of probiotics, let us look at the
details of prebiotics in the next sub-section.

11.5.3 Prebiotics: Dietary Sources and their Mode of Actioflealth

Effects
We have already seen how prebiotics are defined. Let us go a little in-depth about
them. Prebiotic fermentation should favour growth of potentially health promoting
bacteria especially Lactobacilli and Bifirlobactericr: which are indigenous to the
gastrointestinal tract. In a way, the approach is similar to that of dietary fibre or
resistant starch. However, prebiotics will affect bacterial growth selectively. Also,
efficient prebiotics will reduce/suppress the nu~nbersand activities of possibly pathogenic
microorganisms.

Since prebiotics are fermented by the gut microflora, they may have physiological
effects on the GIT such as:
o control of transit time and motility,
regulation of epithelial cell proliferation,
influence on nutrient bioavailability,
modulation of immune function, and
modulation of endocrine function.
In addition, they may exert systemic effect by influencing carbohydrate or lipid
homeostasis.

Our subsequent discussion would be pertaining to soine itnportant types of prebiotics

such as Non-digestible oligosaccharides. Let us read about them.

Non-Digestible Oligosaccharides (NDO)

Among the various food conlponents, the best prebiotic effects seen1 to be exerted
by the NDOs. They are oligomeric cat.bohydrates, which a n resistan't to hydrolysis
by intestinal digestive enzymes but NDOs can be metabolized by colonic bacteria.
What will be the products of bacterial fermentation? Well, these are:
short chain carboxylic acids (short chain fatty acids),
gases; and
organisms also increase metabolic energy, growth and proliferation.
Generally, these carbohydrates are a mixture of oligolners of differing chain lengths.
Oligosaccharides with prebiotic effect are:
o Pnct ooligosaccharides (FOS) or inulin-type fructans
soybean oligosaccharides e.g. stachyose and raffinose
galacto oligosaccharides
o malto oligosaccharides
galactosylsucrose
palatinose condensates
xylooligosaccharides
Advance Nutrition Many prebiotic oligosaccharides are being used in Japan. Among these, FOS have
been the most studied. We will now highlight some interesting features of fructo-
oligosaccharides or fi~lctans.

It is a general term for any carbohydrate in which onelmore fructosyl fi-uctose link
constitutes the majority of glycosidic bonds. They are linear or branched fructose
oligopolymers i.e. P -2,l fructosyl fructose linked inulins or -2,6 linked levans. Inulins
are mainly of plant origin, containing 2 ->70 units, whereas levans are mostly produced
by some fungi and many bacteria. FungaVbacterial inulins have a much higher degree
of polymerisation (DP) (upto 150). Oligofructose (DP 2-10) is produced by enzymatic
hydrolysis of inulin. Thus, as food ingredients, they are available either as native inulin
or high molecular weight inulin, or the enzymatically produced hydrolysate oligofnrctose.

Synthetic fructans are mainly P-2,l linked fructose with DP 2-4, industrially obtained
by enzymatic syilthesis from sucrose. Inulin type fructans are present in significant
amounts in a variety of edible fruits and vegetables e.g. plant families such as
Lilliaceae, Anzaryllidiaceua, Gmmirzae, Conzpositae e.g. asparagus, artichoke, garlic,
leek onion, banana, chicory roots.

Fructans can be ingested by consuming these foods, e.g, in cereals, aerial parts of
many Grarninne particularly young seedlings contain upto 70% of their dry weight.
Estimated daily intake in Western countries is 3-llglday, depending on the type of
food consumed.

Because of their p configuration, inulin-type fructans are resistant to human digestive

enzymes, and in the ileum due to microbial activity and/or enzymatic hydrolysis, with
the remaining 86-88% of the ingested fructans being left undigested.
Isomaltooligosaccharides are partially hydrolyzed by isomaltose in the jejunum whereas
the soybean oligosaccharides raffinose, stachyose and palatinose are not hydrolyzed
much. Galactooligosaccharides appear to be metabolized.

The fi~nctional/technologicalattributes that give them a potential for incorporation into

foods and the health benefits of fructa~sare enumerated next.
A) Health Benefits of Fructarzs
The health benefits of fiuctans include:
BiJ'idogenicEfect: Fr~lctailsselectively stimulate the growth of Bifdobacteria
and Lactobacilli, while decreasing concentrGions of E.coli, Clostridia and
bacteroides. Predominance of these bacteiia is achieved within two weeks, with
effects lasting as long as the fructans are consumed. Daily doses of 4-40 g
increased the bifidobacte~iaupto per gm of faeces.
But what are the benefits of increasing the growth of bifidobacteria?
Bifidobacteria displaces potential pathogens selectively, showing an antibiotic
like effect, which is unrelated to the changes in short chain fatty acids (SCFA)
and pH. Due to fennentation, the carboxylic acids produced-acetate, propionate,
butyrate-have both systemic and physiological effects. Also, these end products
of fermentation allow the host to salvage a part of the energy of NDO and may
play a role in regulating cellular metabolism, cell division and cell differentiation.
Look at Table 11.5, which presents the physiological effects of bacterial growth
and acid produced.
 Table 11.5 : Physiological effects of ferniei~tation Food Components other
than Essential Nutrients
Acid Causes: Bacterial Growth Causes:
decrease in faecal pH increased bacterial bioinass
hyperplasia of intestinal mucosa increased faecal mass
a increased wall thickness in small competitioll effects towards colonization
intestine and cecuin sites and available nutrients
o increased blood flow in the tissue excrete natural anti-microbial agents which
the lowered pH in a micro niche can affect a variety of organisms
makes it unfavourable for pathogens 0 may tl~emselveslunction as anti-infective
agents through occupation of pathogen
colonization or receptor sites

Thus, prebiotics help to attain an appropriate balance of microflora and help in

resisting the effects exerted by the various pathogens.
Mineral absorption: NDOs affect inainly calcium/magnesium absorption and
balance. Acidification of colonic contents increases the concentration of ionized
minerals, particularly calcium and magnesium, thus favouring passive diffusion.
Formation of soluble salts of organic acids and colonic hypertrophy facilitates
increased absorption of these minerals. Similar effects have been observed in
animals (but not yet in humans) for iron and zinc.
o Glycemidnsz~line~~zia: Preliminary research evidence indicates that there may
be beneficial effects in terms of decreased fasting blood glucose/hepatic glucose
production. This may be due to delayed gastric emptying, increased transit time,
which may be dose-dependent. Alternatively, hepatic metabolisin of glucose may
be modified, mediated by the SCFA, especially propionate. Propioilate inhibits
gluconeogenesis probably via its conversion to methylmalonyl CoA and succi~lyl
CoA, both of which inhibit pyruvate carboxylase. Propionate enhances glycolysis
and may lower plasma fatty acid concentrations.
Lipid Metabolism: Aniinal studies consistently show a hypotriglyceridenlic effect
although equivocal results have been seen with healthy humans.
Hypotriglyceridemia is due to decreased VLDL concentrations. Animal studies
show a decreased de novo lipogenesis in the liver due to reduced act,jvity
(almost 50%)of all lipogenic enzymes, possibly through modification of lipogenic
enzyme gene expression. Inhibition is mainly attributable lo propionate. Limited
studies show possible lowering of serum total and LDL cholesterol.
Uremia and Nitrogen Disposal: In animals, faecal and renal nitrogen were
enhanced and decreased uremia was seen in normal and nepl-rectomised animals.
The mechanisms proposed are:
- increased colonic biomass and consequent nitrogen fixation with acidification
and conversion of diffusible ammonia into less diffusible aimnonium ion,
- . due to NDO's osinotic effect, urea transfers into distal ileum and large
intestine is accelerated. In the large intestine, ureolytic rnicroflora may act.
When fermentable carbohydrate intake is high, the amount of ammonia
required may become insufficient and blood urea is used as a substrate, and
- inhibition of ureagenesis in the liver by propionate.
Once again it should be emphasized here that these are inferences drawn from
% animal studies and verification in humans will have to wait until relevant studies are
carried out.
Enumerated below are the potential uses of fructans.
Advance Nutrition B ) Fructnns as Food Ingredients
Oligosaccharides have several functionaVtechnological attributes that give them a
potential for incorporation into foods. They can:
modify freezing point and moisture content,
have variable stability to acid, on storage and processing,
have bactei-iostatic prope ties,
may stabilize proteins,
retain flavour, aroma,
affect colour formation,
variable sweetening power, thus variable energy density and carcinogenicity,
can be used as fat replacers since they confer mouth feel and texture similar
to fat, and
can be used as bulking agents.
Thus, to summarize, we can say r that a good prebiotic should have the following
properties:
a be active at a nutritionally feasible, as low a dose as possible,
lack side effects,
0 be able to exert fine control of microflora inod~~lation,
persist throughout the colon including distal areas,
have varying viscosity,
have good storagelprocessing stability,
have differing sweetness, and
inhibit adhesion of pathogens.
In this section, we learnt about pro- and prebiotics. The discussions highlighted the
different types of these substances, their properties and uses in the food industry. It
must have been interesting to learn about non-digestible oligosaccharides particularly,
fructans. We shall continue to learn further about some more bioactive non-nutritional
components such as polyphenols. However, let us first revise our concepts by
performing the following exercise given in check your progress exercise 2.

Check Your Progress Exercise 2

1) Define the following terms, giving suitable examples.
a) Probiotics
.................................................................................................................
.................................................................................................................
.................................................................................................................
b) Prebiotics
.................................................................................................................
.................................................................................................................
.................................................................................................................
c) NDOs
.................................................................................................................
.................................................................................................................
 Food Components other
2) Enlist the characteristics of Probiotics and Prebiotics. than Essential Nutrients
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
3) What is the possible mechanism by which probiotics act?
.....................................................................................................................
........................................................................................................i..L..,. .....
.....................................................................................................................
4) Describe any two physiological effects of prebiotics and probiotics.

.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
5 ) Discuss the benefits attributed to the intake of NDOs.
.....................................................................................................................
.....................................................................................................................
....................................................m ...............................................................
6 ) What characteristics of Fructans give them a potential for incorporation into
foods?

.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
1

11.6 POLYPHENOLS
We now come to a group of compounds, which were until recently considered to
adversely influence nutrient absorption or giving colour to foods e.g. anthocyanins.
Today they are among the most popular and commonly consumed nutraceuticals.
More th'ul 4000 phenolic phytochemicals have been identified so far including
tocopherols and tocotrienols. They are ubiquitous in all plant organs and therefore are
an integral part of the human diet. Let us find out what are these.
 Advance Nutrition 11.6.1 Definition and Glassif'ication
Polyphenols are the compounds that contain an- ON group attached to the
benzene ring. The main classes are: flavonoids, phenolic acids and polyphenols
(tannins). They arise from common intermediates pllenylalanine or its precursor shikin1ic
acid. Natural phenolic compounds range from simple ones like hydroxybenzoic acid
to highly polymerized ones e.g. tannins. They arc glycosylated by one or more suga
residues. The OH groups linked can be glucose, galactose, rhsunnose, xylose or
arabinose, glucuronic or galacturonic acids.
The various classes are:
a) Phenolic acids and derivatives
b) Flavonoids such as Flavonols and Flavones, Isoflavones and Flavanols
C) Tannins
d) Stilbenes

We will now describe each of these one by one.

. A) Phenolic Acids and Derivatives

l-wo families of phenolic acids are widely distributed in plants: a range of benzoic acid
derivatives and those deiived from cinnamic acid, both occul-rillg in a conjugated or
an esterified folm. Examples are:
e Hydroxybeizzoic acid: e.g. ellagic and gallic acids, which are hydrolyzable
tannins, present in berries and nuts.
e HyLZroxycinnarnic acid: e.g. caffeic and fenllic acids, which are heat-sensitive.
Caffeic acid is a precursor of lignin. Caffeic wit11 qunic acid gives chlorogenic
acid. They occur in fruits, vegetables notably seeds, coffee beans, grains,
sunflower seeds. Among these, curculnin and chlorogenic ncid have been sh~died
more. Anticarciiiogenic effects are attributed to caffeic and ferulic acid. They
may act in two ways:
a) prevent formation of carcinogens froin precursors, and
b) block reaction of carcinogens with critical cellular macroinolecules.
Next, let us get to know about the flavonoids.
B ) Flavonoids
Flavonoids constitute the largest group of plan1 polyphenols, generally arc the compo~lnds
of low molecular weight, bound to sugar molecules. Yo11 may be familiar with some
of these plant pigments. Antlzocynrzirzs (red, blue and purple pigments) are flavanoids.
We also have aizthoxanthirzs, whicl; include flavonols, tlavoncs, flavanols
and isoflavones. All of these are colourless/white/yellow. We will look at each
briefly:
Flavonols and Flavones: These are the most widely distributed slmoilg the
flavonoids. The most commonly occurring are Quercetin Myricetin and
Kaempferol. Quercetin is quantitatively the most important and is present in
onions, apples, kale and tea. It is concentrated in the outer exposed parls of
plants and its concentration is proportional to the greenness. Losses V L ~ average
;

cooking methods result in less than 20% losses whereas other processi~igmethods
lead upto 50% losses.

368
 c Flavanols: Examples are catechin and epicatechin. You may have heard of Food Components other
than Essential Nutrients
the benefits of tea. Some of them are due to gallic acid, which is combined with ~
I

epicatechin. Flavonols constitute 30% of dry matter from tea solutions. In fruits,
legumes and grains, they exist as condei~sedpolyil~ersand are concentrated
more in immature fruits. Fruits stored over winter lost approxin~atelyhalf their
content.
0 IsofZnvorzes: These are usually treated separately from the other five s~~bclasses
and are an area of considerable research interest. Isoflavones are found almost
exclusively in the legume family but occur in high amounts only in soybeans.
Genistein and Daidzeirz are found in soybeans. They are heat stable, slightly
water-soluble. The content in dry soybeans ranges from 1600-2400 m g k g .
The third polyphenol covered in tlis section are tannins.
C) Tannins
Long known for their inhibitory effects on iron absorption, recent research indicates
that tannins do have beneficial effects. Tannins are compounds of high molecular
weight. In fact, the astringent taste that you find in some foods is due to the reaction
of tannins with mouth proteins. Figure 11.1 highlights some of their characteristic
features.
Tannins are

Condensed Ilydrolyzable
0 polymers of catechins/epicateclli~ls contain a central core of polyhydiic
0 structurally more complex than alcohol e.g. glucose or hydroxyl groups
hydrolyzable tannins polymers (esterified partially or
referred to as 'Leucoa~~~l~ocyanidi~~s' wholly) of gallic/ellagic acid
because they form cyanidin upon acid found in berries/nuLs
hydrolysis call be easily hydrolyzed by acids,
found mainly in fn~its,grains, legumes bases or certain enzynles
usually accumulate in outer layers of
plants
0 semoved by dehulliilg or heating in
water
not degraded by acid treatment,
but polymerize
?

Figure 11.1: Characteristics of different forn~sof tannins

D ) Stilbenes
Stilbeizes contain 3 phenyl colnpounds connected by a 2-carbon methylene bridge
and occur in nature in restricted distribution. In plants, they act as anti-fungal
phytoalexins, usually synthesized only in response to infection/injury. T h e most
extensively studied is trans-reservatrol, present in grapes, wine and peanuts. Red
wine contains 1.5-3.0 mg/L.
Finally, let us get to know the lignans.
E) Lignans
Lignans are diphenolic compounds formed by dilnerization of 2 cinna~nicacid residues.
Most lignans apparently pass through the GIT as fibre. Some lignans may be converted
by intestinal microflora to mammalian lignans-enterodiol and enterolactone, which are
absorbed through enterohepatic circulation. Flaxseed meal and flour are said to be the
richest sources of 2 lignans - Secoisolariciresiizol diglycoside ( S D G ) a n d
matctiresinol. Other good sources of lignans are pumpkin seeds and sunflower seeds.
Next, we shall read about the absorption/bioavailabilityof polyphenols.
Advance Nutrition 11.6.2 Bioavailability of Polyphenols I

What do you mean by the term 'bioavailability'? You would recall reading about it
earlier in this course. We suggest you to go back and refer to its definition in Unit
7 and 8 and then proceed. further.

We need to consider bioavailability before studying the functions because their

nutritional significance depends on their behaviour in the digestive tract. Some of the
salient aspects related to bioavailability have been elucidated herewith:

i) their digestion and utilization depends on their structure, degree of glycosylationl

acylation, conjugation with other phenolics, molecular size and stability,
ii) in vitro studies and animal experiments show that upto 113"' of extractable
polyphenols are excreted, however, the percentage o'f non-extractable polyphenols
excreted are twice or three times more,
iii) some phenolics like aglycones, simple phenols and flavonoidsare directly absorbed
into the small intestines. Glycosylated polyphenols are hydrolyzed by glycosidases
present in food/produced by the GIT mucosa or they are acted upon by microflora,
after which they are absorbed in the colon. Intestinal bacteria have been found
to solubilize non-extractable polyphenols, and
iv) absorption in rats is low (20-34%). In l ~ ~ ~ m athe
n s , absorption is apparently
influenced by the chemical nature, thus the extent varies from as much as 24-
52% for Quercetin, 9-21% for isoflavones to as little as 0.2-0.9% for tea
catechin.
Human studies on bioavailability indicate that plasma levels of flavonoids are close
to 0.55 pmol/L but do not exceed 1 pmol/L when the dose was close to dietary
intakes.
With a brief understanding about the bioavailability of polyphenols, next we shall study
the influence of polyphenols on ~llacronutrientsand rniperals.

11.6.3 Influence of Polyphenols on Macronutrients and Minerals

Let us study in this sub-section how the polyphenols affect the digestion and absorption
of various macronutrients such as proteins, carbohydrates, lipids and minerals.
A) Proteins
You know that tannins are considered as anti-nutritional because their presence is
usually accompanied by a reduced protein digestibility and a subsequent increase in
the faecal nitrogen. Decreased protein digestibility may be due to the formation of
'protein-tannin complex', as well as, inhibition of proteolytic enzymes like lrypsin by
polyphenols. Polyphenols have exhibited a highly significant and negative correlation
with in vitro digestibility of proteins.
However, this may have a beneficial effect in overweight or obesity, since tannins
bind to endogenous proteins in the intestinal tract such as digestive enzymes like
lipase and amylase and inhibit them, resulting in lower energy availability.
B) Carbohydrates
Binding of proteins may indirectly affect carbohydrate absorption. Inhibition of :
amylolytic enzymes and subsequent reduction of dietary carbohydrate hydrolysis can
decrease the postprandial glycernic response, which can have a beneficial effect for :
diabetics.
C) Lipids
The effect of food polyphenols on lipid metabolism has not been extensively studied.
Both soluble polyphenols and condensed tannins have been shown to decrease the r
1
apparent digestibility of lipids, lipase activity and increase faecal fat excretion. E
D ) Bionvailability of Minerals Food Components other
than Essential Nutrients
Polyphenols can form complexes with metal cations through their carboxylic or
hydroxylic groups, and thus interfere with the intestinal absorption of minerals like
iron, copper, sodium and aluminum. The important negative interaction in the Indian
setting is the iron absorption vs. polyphenols, as Indian diets are deficient in iron.

In view of the impact of polyphenols on the metabolism of several macro and micro
nutrients, research has been carried out to explore their potential in the treatment,
management and prevention of various diseases. Enumerated next are some of their
health benefits.

11.6.4 Health Benefits of Polyphenols

Various health benefits have been attributed to polyphenols. These include:
- anti-microbial
- anti-viral
- anti-oxidant
- hypotensive
- hypoglycemic
- anti-carcinogenic
- anti-mutagenic
- oestrogenic
- anti-ulcer .
- anti-inflammatory
- prevent expression of adhesion molecule
- inhibit replication of HIV
Most of these propelties (health benefits) are due to their anti-oxidant activity. Both
in vitro and in vivo studies have been conducted to elucidate these effects. Let us
read further Lo understand the various properties of polyphenols in detail.
A) Antioxidant Action of Polyphenols
The properties of polyphenols due to which they have been identified to act as
antioxidant is because they:
- act as terminators of free radicals by rapid donation of H atoms,
- chelate metal ions like iron and copper,
- act as terminators of propagation,
- quench reactive oxygen species and reactive Nz species,
- inhibit lipoxygenase and cyclo-oxygenase enzymes, and
- inhibit damage by haernprotein/peroxide mixtures.

The antioxidant function depends on the number of hydroxyl groups and their positions.
Among the various derivatives, flavonoids are the most potent and retain their
scavenging capacity even after chelation with the metal ions. Non-extractable
polyphenols are apparently 15-30 times more effective than simple phenols. A point
to note is that their ability to chelate metals also makes them act as pro-oxidants-in
situation when tissue injury causes release of iron/copper.

The most important antioxidant role of polyphenols identified so far has been with
respect to the treatment, management and prevention of several degenerative diseases
particularly, heart diseases and cancer. We will read about these in detail.
 Advance Nutrition B ) Cardiovascular Effects
You must have heard of the 'French Paradox'. The lower incidence of heart attacks
in certain areas of France despite higher prevalence of factors like smoking and high
fat intake was attributed to the red wine consumption in these areas. Subsequently,
the 'Zutphen study' in the Netherlands showed an inverse relation between the
incidence of CHDI stroke in elderly men with the dietary intake of flavonoids especially
quercetin, which originated mainly from tea, fruits and vegetables. Average total daily
consumption of flavonoids was at least 23 mg (16 mg quercetinlday) and the average
vitamin E intake was 7-10 ing.

Phenols have been found to consistently decrease several risk factors/CVD

indicators - plasma LDLc was less susceptible to oxidation, contained lower amount
of lipid peroxides. There was lower plaque formation in spontaneously hypertensive
rats, consumption of wine polyphenolic extracts improved aortic blood pressure and
vessel mechanical properties.

Next, we move over to the prevention and inhibition of cancer.

C) Cancer and Inlzibition of Tumorigenesis

Polyphenols appear to play a preventive role although the molecular mechanisms of
action and applicability to human cancer prevention are unclear. Various studies
appear to suggest that they may be effective when given in the post initiation period
i.e. they are more helpful in inhibiting tumor promotion and progression.

Most studies have been conducted either in vitro or in vivo in animals. Hence, we
must be cautious in extrapolating these observations to humans. The observations
from studies are summarized as:

i) caffeic acid, ferrulic acid, chlorogenic acid and curcumin inhibited tumor promotion
-curcumin inhibited colon tumorigenesis.
ii) multiplicity of invasive and non-invasive adenocarcinolna was lower.
iii) apoptosis of colonic tumor cells was higher.
iv) expression of oncoprotein was decreased.
V) in mammary tumors, these compounds cause inhibition directly or by modulating
oestrogen action.
These effects have been observed with tea catechins and related compounds, caffeine,
quercetin and other flavonoids, isoflavones and lignans, reseveratrol and other grape
constituents. How do they confer beneficial effects? Well, this is through the following
mechanisms:
inhibit activation of carcinogens,
facilitate elimination of certain carcinogens or their reactive intermediates,
inhibit the metabolism of arachidonic acid-decreased production of pro-
inflammatory or mitogenic metabolites like certain prostaglandins or reactive
oxygen species,
modulate hormone-dependent carcinogenesis,
modulate different oncogenes, suppressor genes, signal transduction pathways.
Thus, they inhibit cell proliferation, transformation and induce apoptosis, and
promote terminal differentiation of human tumors.

It must be evident to you by now as to why polyphenols have become popular

nutraceuticals world wide and are being consumed frequently as an over the counter
food supplement. Let us now move on to the next group of compounds, phytoestrogens.
372
But before that, you need to quickly review what we have learnt till now by answering Food Components other
the questions included in the check your progress exercise 3. than Essential Nutrients
- -

Check Your Progress Exercise 3

1) What are Polyphenols? Name the compounds from which they arise.

.....................................................................................................................
2) List the salient aspects of bioavailability of polyphenols.

.....................................................................................................................
3) Briefly discuss the influeilce of Polyphenols on:
a) Minerals

b) Macronutrients

..............................................................................................................
4) Enumerate any five important health benefits of polyphenols.

5) What characteristics of polyphenols are responsible for their anti-oxidant action?

.....................................................................................................................
Advance Nutrition
6 ) Match the following:
A B
i) Phenolic Acid a) Genistein and Daidezin
ii) Flavonols b) SDG and Matairesinol
iii) Isoflavones c) Hydroxybenzoic acid, Hydroxyannamic acid
iv) Lignans d) trans-reservatrol
v) StiIbenes e) Catechin and epicatechin

1 . 7 PHYTOESTROGENS
You may be aware of the surging interest in phytoestrogens (PE) especially in
connection with osteoporosis. This section briefly discusses these compounds.

As the term implies, they are the oestrogenic compounds found irz plants. They
exert oestrogenic effects on the central nervous system, induce estrus and stimulate
growth of the genital tract of female animals. In a broader sense, they are chemicals
showing effects suggestive of oestrogenicity e.g. binding to oestrogen receptor (ER),
inducing specific oestrogen-responsive gene products and stimulating ER-positive
breast cancer cell growth.

Phytoestrogens can be divided into 3 main classes: isoflavones, coumestans and

lignans. All are diphenolic compounds with structural similarities to natural and
synthetic oestrogens and anti-oestrogens. Our subsequent discussions highlight the
major forms of phytoestrogens and their food sources.

11.7.1 Dietary Sources and Chemical Forms

Isoflavones and coumestans are the most common compounds. Soybeans and
soyfoods are the most important sources, containing approx. 0.2-1.6 mg per gram dry
weight; although chick peas and other legumes also contain them. Second generation
soy foods e.g, products like tofu yoghurt contain 6-20% of the isoflavones present
in the whole beans.

Isoflavones may be present in unconjugated form (aglycones) e.g. daidzein, genistein,

or as glycosides such as daidzin, genistin.

Non-fermented soy foods like tofu contain greater levels of glucosides whereas
fermented foods e.g. tempeh have higher amounts of aglycones due to enzymatic
hydrolysis during the fermentation process.

A large number of cournestans e.g. coumestrol in alfalfa have been isolated from
plants but only a few possess uterotropic (stimulating growth of the uterus) activity.
The highest amounts of coumestans are present in clover and alfalfa sprouts (5.6 and
0.7 mglg dry weight). Other sources are split peas, kala chana, lima beans and soy
bean sprouts (15-80 mcg/g dry weight).

Lignans do not induce estrus but are considered to be phytoestrogens because they
have oestrogen-like actions. They are present in plant foods and human biological
fluids, plant lignans being converted by bacterial action to mammalian lignans in the
GIT. Secoisolariciresinol and rnatairesinol are plant dietary precursors of the mammalian
lignans, enterodiol and enterolactone.

The content of lignans in various foods is shown in the Table 11.6, oilseeds being the
richest plant sources. 9
 Table 11.6: Lignan content of selected foods Food Coniponents other
than Essential Nutrients
Plant Source Content ( mg per 100 g wet wt)
Flaxseed 0.8 mg secoisolariresionl per g dry wt
Unhulled soybeans 20.5
Dried sea weed 0.9
Whole legumes 0.6
Cereal brans 0.5
Legume hulls 0.4
Whole grain cereals 0.3
Vegetables 0.14
Fruits 0.03

The potential role of'phytoestrogens with respect to maintaining good health is being
discussed next.

11.7.2 Physiological Effects

Little is known about the biologic and physiologic effect of phytoestrogens in humans.
Animal studies suggest the following physiological effects of phytoestrogens:
hormonal effect, like stimulation of uterine growth in a manner similar to oestrogen
(whether phytoestrogens have oestrogenic or anti-oestrogenic effect will depend
on the amount of endogeneous oestrogen present), and
e antioxidant effects by increasing the activities of antioxidant enzymes such as
catalase superoxide dismutase, GSH peroxidase and GSH reductase. Soya
isoflavones may also act directly as anti-oxidant.
The health benefits ascribed to phytoestrogens are for protection against cancer,
relieving symptoms of menopause like hot flushes, prevention of osteoporosis (through
prevention of bone resorption and promoting increased bone density) and prevention
of heart disease in postmenopausal women probably acting as oestrogen agonists
(mimicking the effect of oestrogens), producing effects on lipoproteins similar to
those caused by oestrogen.

Let us now look at the characteristic features of other food components with
antinutritional effects. You may be familiar with the terms phytate and trypsin inhibitor.
Let us discuss these and others.

11.8 OTHER DIETARY FACTORS WITH

ANTINUTRITIONAL EFF'ECTS
In the above sections, we have learnt about a variety of food components having a
host of health promotive properties. Let us now have a look at a few factors with
antinutritional effects. These factors in foods include protease inhibitors physic
acid, phenolic compounds, amylase inhibitors and saponins. The well known anti
nutritional effects of these factors are discussed first. These factors are present in
foods that we consume regularly such as whole wheat, other whole grains, legumes
and soyabeans. A few noteworthy points: The concentration of antinutritional factors
is much lower in the refined cereals from which nearly dl the bran and much of the
endosperm has been removed. Further, some of the anti-nutritional factors such as,
protease inhibitors in soyabean and some legumes, are heat labile and therefore
proper cooking will inactivate them. Household processes such as soaking and
fermentation will also remove many of the antinutritional factors. The traditional
methods of cooking and processing all have the effect of reducing the antinutritional
factors in our diets. Therefore. YOU will notice that although several adverse effects
Advance Nutrition are documented chiefly in animal models for these antinutritional factors, in actual
practice, the cooking and processing of foods for human has evolved in such a way
as to keep these antinutitional factors low in our diets so that we do not suffer from
the ill effects. Occasionally, however, the consumption may reach levels at which ill
effects become manifest. It is important, therefore, to be aware of these adverse
effects. Following the adverse effects of each of these constituent, the currently
known beneficial effects are listed. At the current time what we do not know is the
amount of these to be consumed to maximize the beneficial effects and minimize the
adverse effects.

So then let us begin our study of these factors with protease inhibitors.

1 1 . 1 Protease Inhibitors
These are protein in nature and are abundant in raw cereals and legumes, especially
soybeans. It would be interesting to note here that since they are proteins, they can
be denatured by heat. However, in commercial soy products, 5-20% of the activity
has been detected.

What are the effects of protease inhibitors?

In experimental animals, they have been found to be associated with growth inhibition
and pancreatic hypertrophy. A negative feedback mechanism in the small intestines
has been postulated for these effects. In the presence of trypsin inhibitors, iqactivation
and loss of intestinal trypsin can take place. These can trigger the release of
cholecystokinin (CCK) from the intestinal mucosa. CCK then can induce the pancreas
to produce more trypsin. However, excess of hypsin inhibitors can reduce the protein
available thus explaining the growth inhibition seen in animals, as well as, the pancreatic
hypertrophy. Moreover, trypsin is rich in sulphur-containing amino acids; hence increased
synthesis of trypsin (the more the inhibitor, the more trypsin that is made) increases
the requirements for these amino acids, ultimately leading to weight loss. Also, the
stress of the pancreas results in pancreatic hypertrophy and hype~plasiaof the acinar
(exocrine) cells.

Feeding purified trypsin inhibitor or raw soy flour containing protease inhibitors
potentiated the effects of pancreatic carcinogens. It must be noted that most of
the effects have been observed in some species of animals only, whose pancreas
constitute a fairly higher percentage of body weight (0.29-0.8%) e.g. in rats, mice,
chickens, hamsters and guinea pigs but not in large animals with a small pancreas
(0.06-0.24% of body weight) e.g. dogs, calves, monkeys.

In human subjects, provision of raw soy flour or purified trypsin inhibitor directly to
the duodenum, increased secretion of pancreatic enzymes and serum level of CCK.
This suggests that a negative feedback mechanism also exist in humans. However,
it is interesting that among populations with fairly high intake of soybeans or other
foods rich in protease inhibitors, e.g. in Japanese and Seventh Day Adventists, the
incidence of pancreatic cancer is lowei.

Another category of food components which may influence human health are the
saponins. A few characteristic features of saponins are discussed below.

11.8.2 Saponins
These are a diverse group of compounds commonly found in legumes like soybean,
lentils, clickpeas, peanuts and alfalfa sprouts. They are also present in some plants
which we use as flavouring agents such as herbs and spices e.g, fenugreek, nutmeg,
ginseng, sage and thyme.
Let us briefly understand their chemical nature. Food Components other
than Essential Nutrients
Their structures are characterized by the presence of a steroid or triterpene group,
i.e. an aglycone linked to one or more sugar molecules. Since these compounds have
both polar (the sugar moiety) and non-polar (steroid or triterpene) group, obviously
they will have surface-active properties. These properties are responsible for whatever
effects saponins exert.

You should rerneinber that saponins have both adverse, as well as, beneficial biological
effects. Let us first list the adverse effects.
The adverse effects of saponins may be described as under:
- A well-known toxic effect is 'erythrocyte lysis' since they interact with the
cholesterol in the erythrocyte membrane.
- In mammals, intravenous adnlinistration causes local inflammation.
- In large doses, it results in death due to ~nassiverelease of erthyrocyte debris.
- A reduction in oxygen-carrying capacity of blood.
- It can lyse other cells e.g. of the intestinal rnucosa and thus affect nutrient
absorption.
- High saponin intake results in decreased weight gain. This has been attributed
to a number of reasons:
bitter taste
decreased nutrient absorption and utilization because of inhibition of metabolic
and digestive enzymes such as amylase, lipase, protease and cholinesterase.
These effects have been observed with soy saponin.
Alfalfa sprouts have been found to inhibit chymotrypsin, protease, succinoxidase, as
well as, bind zinc.

Now let us study the beneficial effects of saponins. Saponins may have some
anticarcinogenic effect by binding primary bile acids, formation of secondary bile
acids and thus the promotion of tumorigenesis is reduced. They also enhance the
immune response and have been used as adjuvants for oral vaccines.

Another group of substances of interest are the commonly consumed amylase

inhibitors. These are most frequently consumed in view of the fact that they are
present in several cereals and legumes and we know that Indian dietaries are principally
cereal based.

11.8.3 Amylase Inhibitors

Inhibitors of the enzyme a-amylase are found in wheat, rye, beans, mango, legumes,
potatoes, sorghum (jowar) and oats. Most amylase inhibitors from plants are active
against animal amylase.

Let us see how these inhibitors act.

The inhibitor forms a complex with amylase, which in turn reduces starch digestion.
Complex formation is influenced by pH, ionic strength, temperature, time of inactivation
and inhibitor concentration. Other effects observed are pancreatic enlargement and
hyperplasia.

When starch digestibility is reduced and in case starch intake is limited, you may
expect growth to be adversely influenced. However, observations on animals provide
conflicting data. It is possible that amylase inhibitors from different sources may have
Advance Nutrition different effects due to pH sensitivity and sensitivity of the pancreas to different
amylase inhibitors.

In humans, slightly different effects have been observed e.g. diarrhoea, nausea and
vomiting. It is noteworthy that these effects were observed after intake of 'starch
blockers' which may not be pure amylase inhibitors and other antinutritional factors
like lectins and protease inhibitors also caused some of these effects.

Yet another category of food substances which are not conducive to good health
include lectins and haemagglutinins. These terms may be relatively new to you. Let
us understand some of their salient characteristic features.

11.8.4 Lectins or Haemagglutinins

These are sugar binding proteins, having the ability to bind and agglutinate red blood
cells (RBCs). You will find that they occur in most plant foods including those, which
may be eaten raw. Lectins are specific in the sugar they bind to and also in their
toxicity. Of great interest would be the fact that lectins froin horse gram, lima beans,
kidney beans, mung, winged beans and castor beans are toxic when orally consumed
but lectins from soybeans and peanuts are not.

What are their toxic effects?

Well, these can cgbse growth inhibition in animals and diarrhoea, nausea, bloating and
vomiting in case of human beings. When injected lectins agglutinate RBCs, haemolysis
and death occurs in extreme cases. Several outbreaks in England after consuming
improperly cooked beans have been related to lectins although other antinutritional
factors may have played a role. Since lectins are proteins, they are denaturated by
heal: however, low temperature or slow cooking may not completely eliminate their
toxicity.

Toxic effect of lectins also relates to their binding with specific receptor sites on
intestinal mucosal cells. This results in lesions, disruption and abnormal developmei~t
of microvilli. Other effects include reduced activity of brush border enzymes viz.
peptidases, disaccharidases, alkaline phosphatase, as well as, pepsin and pancreatic1
salivary amylase. Secretion of mucin and increased weightfnumberof intestinal mucosal
cells in presence of lectin has also been observed.

Since the enzymes secretionlactivity are affected, carbohydrates and proteins are not
completely digested. When these substances reach the colon, they act as substrates
for the colonic microflora, which subsequently ferment them and produce short chain
fatty acids and gases. This is the underlying reason for some of the gastrointestinal
symptoms associated with intake of raw beanslpurified lectin.

Bacterial overgrowth may also occur and may contribute to lectin toxicity. This has
been linked to thinning of the jejunal mucosa and damage to duodenal.microvilli.
Increase in bacterial colonization by lectin may be due to its polyvalent nature, which
allows lectin to bind to both mucosal cells and bacteria simultaneously. This would
'fix' the bacteria close to the intestinal mucosa. If the mucosa is disrupted, it may
allow translocation of the bacteria andfor endotoxin and cause a toxic response.

In addition, there are reports that lectins themselves may be internalized, causing
systemic effects such as increasing protein catabolism, breakdown of stored glycogen
and lipid, as well as, disturbing mineral metabolism.

Yet another type of food components which may influence nutrient absorptionhealth
include phytates. You must have read about them before especially with respect to
iron absorption. We will now discuss some salient features of phytates.
 11.8.5 Phytates Food Components other
than Essential Nutrients
We are familiar with phytates as an inhibitor of mineral absorption (calcium, iron etc.)
especially in the vegetarian diets that are cereal-based. This sub-section deals with
the well-known nutritional implications which have been long researched, as well as,
the health benefits, which have been in focus more recently.
Let us begin our study with the chemistry and occurrence of phytates.

Chemistry and Occurrence

Most plant foodstuffs contain inositol which is present in the form of inyo-inositol
hexakisphosphate i.e. phytic acid and its salts. In seeds, inositol is the storage form
of phosphate and 60-90% of phosphate is present as phytate. Phytic acid (PA) is
generally complexed with nlincrals and/or proteins. The complexed fonn with minerals
is known as 'Phytate'. Various cations can strongly chelate between two phosphate
groups or weakly within .a phosphate.

Our subsequent discussions will elucidate the interaction of phytates with nutrients,
as well as, their health benefits. We will begin with influence of phytates on mineral
absorption.

Nutritional Implications
We have already discussed about the influence of polyphenols on the various
macronutrients and minerals earlier in Unit 9 and 10. Let us in this sub-section once
again review the nutritional implications of these components.
Phytate and Mineral Interaction: Phytic acid occurs as a crystalline globoid inside
protein bodies in the bran of cereal grains. It is negatively charged over a wide pH
range. This will enable it to react with the positively charged ions. Thus, PA binds
to mineral cations such as Ca2+,Fez"', Fe", Zn 2+ and Cu2+ and forms strong
complexes which render thein ~mavailablefor absorption. The effects of PA on
mineral availability depends on numerous factors:
the amount of PA present,
a concentration of the minerals,
a molar ratio of PA to minerals,
size and valency,
association of PA with protein,
pH,
a presence of other metal ions,
presence of other substances which can bind minerals, e.g. fibre, tannins, oxalic
acid, and
processing methods used.
At the pH values between 5 and 7 which are similar to those in the duodenum, PA
binds and form insoluble chelates with divalent cations. At pH 7.4, the order of
complexation in decreasing order is Cu > Zn > Co > Mn > Fe> Ca.
As phytates accumulates in various storage sites, other minerals apparently chelate
to it forming the complex salt 'phytate'.
.
4
'

Studies on animals show that phytate inhibits zinc and calcium absorption, the maximum
inhibitory effects being shown by 'myoinositol hexaphosphate.' The inhibitory effects
appear to be greater for Zn than for calcium. It appears that at least 5 of the 6
possible sites need to be phosphorylated in order to exert its inhibitoiy effect. In vitro
studies on availability of iron and zinc from North Indian diets indicate that it is very
low-3.3 to 4.4% for iron and 7.8 to 8.7% for zinc, with a negative correlation
between availability and PA content.
Advance Nutrition Next, we will discuss the influence of phytates on protein metabolism.

Plzytate and Protein Interaction: You may be more familiar with phytate's role in
mineral absorption. However phytates also bind with starch and protein. Depending
on the pH, 2 types of complexes are formed-at acidic pH, a binary protein-phytate
complex and as the pH approaches neutrality, a ternary protein-mineral-phytate
complex. Strong electrostatic attraction between negatively charged phytates and
proteins with a net positive charge results in the formation of protein-phytate complexes.

Apparently most such complexes are formed de novo in the gastrointestinal tract.
Also phytates can interfere with digestive enzymes and their substrates, thus adversely
affecting digestion of dietary protein and increasing endogenous losses of amino
acids.

With this, we end our discussion of phytates. Our study on other factors with
antinutritional effect, shall not be complete without a brief mention about the health
benefits of these factors. Yes these factors can be beneficial to health. Let us quickly
review the health benefits.

11.9 HEALTH BENEFITS OF OTHER DIETARY

FACTORS WITH ANTINUTRITIONAL EFFECTS
Despite the predominantly nutritional antagonistic effects of the factors described
above, there is some evidence accumulating to show that these may be associated
with some health benefits as well. Some of the benefits noted with these factors
appear to be similar to those observed for fibre e.g. lowering blood glucose/blood
lipids and lowering the risk of developing cancer. These are briefly reviewed below.
Hypoglycemic effects: Of the different carbohydrate foods tested for starch
digestibility and blood glucose response, those rich in anti-nutrients e.g. legumes
show the lowest values. Effects were best seen with the soluble whey fraction,
which is rich in anti-nutrients like tannins, lectins and phytic acid. Amylase
inhibitors can reduce blood glucose and raise insulin levels.
Reduction of blood lipids: This effect have been especially observed with
saponins. Diets containing saponin-rich foods (300-500 mg saponins/day) e.g.
soya, alfalfa, chickpea, bean meal reduced plasma cholesterol by 16-24%.
However, other substances in these foods like phytosterols, isoflavones, protein
and fibre may also have contributed. Cholesterol reduction may be due to
saponin binding dietary cholesterol thus preventing its absorption andlor binding
with bile acids and increasing their faecal excretion.
Reduction in cancer risk: Different cancers, especially colon and breast cancer,
has been linked not only to phytates but also to protease inhibitors. In vitro
studies show that they suppress malignant transformation of cells. Among the
protease inhibitors, the most effective are those with chymotrypsin inhibitor
activity in foods like soybean, chickpea and potato. Soybean also has the Bowman-
Birk inhibitor which inhibits/prevents development of chemically induced
liver cancer.
What are the mechanisms possibly responsible for these eflects? I

There are several possibilities: reduced availability of amino acids to cancer cells by
reversing the carcinogen-induced change in oncogene expression, or by inhibiting
formation of superoxide radicals, hydrogen peroxide formation by neutrophils which ,
is induced by tumor promoters.

:
Research has indicted that phytates inay have some beneficial aspects as well. Let Food Components other
than Essential Nutrients
us le'arn about them.

Benejcial effects of phytates: Soine of the health promotive aspects of phytates

include:

1) Phytic acid is a known antioxidant. Colonic bacteria produce oxygen radicals in

appreciable accounts. Dietary phytic acid suppresses free radical mediated
damage to intestinal epithelium.
2) Protective benefits are seen particularly in relation to carcinogenesis, particularly
against colon cancers. Complexing of iron by phytate may reduce iron-catalyzed
production of free radicals and reduce lipid peroxidation. Also, phytate-containing
foods have a higher fibre content. Studies show that signal-transduction pathways,
cell cycle regulatoiy and differentiation genes, oncogenes and tumor-suppressor
genes may be involved in phytate's anti-neoplastic' effects.
3) Lowering blood glucose and lipids.
4) Binding amylase and reducing starch digestion.
5) Lowering plasma cholesterol and triglycerides.
In the last two section(s) we read'about cei-tain food components such as protease/
amylase inhibitors, saponins, lectins and phytates, which are at times referred to as
antinutritional factors. These food coinponents have been found to inhibit nutrient
absol-ption and a few substances also are healtl~promotive. Research however needs
to be conducted to further ascei-tain their health benefits especially with regards to
their type and dosage. We hope that it must have been a good learning experience
for ybu. A list of references has been mentioned ar the end of the course for you
to learn further.

You may now need to perfoim the check your progress exercise 4 to recapitulate the
details of the food compoilents discussed in this section.

Check. Your Progress Exercise 4

1) What are phytoestrogens? What are its classes and give exa~nplesof each
class.
......................................................................................................................
.....................................................................................................................
.....................................................................................................................
2) Enlist the food sources of lignans.
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
3) Discuss the toxic effects of:
a) Saponins
..............................................................................................................
..............................................................................................................
b) Lectins
..............................................................................................................
..............................................................................................................
.
Advance Nutrition
4) Enumerate the health benefits of antinutritional factors.
................................................................................................................
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................
.....................................................................................................................

11.10 LET US SUM UP

In this unit, we studied about the health benefits adverse effects of several food
components apart from the nutrients. We got to know about the concept of functional
foods and which components make them healthier food choices. Subsequent discussions
highlighted the characteristic features of several nutraceuticals/bioactive substances/
food components such as probiotics, prebiotics, non-glycoride components of edible
oils, phytoestrogens etc. Their mode of action, potential beneficial health effects food
sources and clinical uses have been elucidated in this unit. We also saw their interactions
with nutrients such as proteins and minerals, especially in the case of phytates.
In the subsequent sections, we learnt that the physiological effects of the various
substances will depend on their level of intake, presence of other dietaiy constituents
and the nutritionalhealth status. Benefits and risks need to be balanced. However,
considerable work is required before we can issue recommendations in terms of
amounts of these to be consumed for the population at large.

11.11 GLOSSARY
Adenocarcinoma cancer that begins in cells that line certain
internal organs and that have secretory
properties.
Angiogenesis the growth and proliferation of new blood
vessels.
Apoptosis one of the types of programmed cell death
that is the deliberate suicide of an unwanted
cell in a multicellular organism.
Fructans a general telm for any carbohydrate in which
one/more fructosyl fructose link constitutes
the majority of glycosidic bonds. I
Functional foods the food substances beyond the basic
nutrients th4t are designed to lower the risk I

or delay the onset of certain disease.

Lectins sugar binding proteins, having the ability to

Non-digestible oligosaccharides :
bind and agglutinate RBCs,
oliogomeric carbohydrates which are
I
resistant to hydrolyzed by colonic bacteria.
Phytate complexed form of phytic acid with minerals.
Phytoestrogens oestrogenic compounds .found in pl'ults. 1
1
Polyphenols compounds containing an -OH groups I
I
attached to the benzene ring.
Prebiotics : non-digestible food ingredients that Food Components other
beneficially affect the host by selectively than Essential Nutrients
stimulating the growth and/or activity of
one or a limited number of bacterial in the
colon.
Probiotics : a live illicrobial feed supplement which
beneficially affects the most animal by
improving its intestinal microbial balance.
Tumorigenesis : formation or production of tumors.

111.12 ANSWERS TO CHECK YOUR PROGRESS

EXERCISES
Check Your Progress Exercise 1
I) The food substances, beyond the basic nutrients that are designed to lower the
risk or delay the onset of certain diseases are called the functional foods. They
are found in mixed culture of living organisms, many fluits, vegetables, grains,
herbs and spices.
2) As per the Japanese criteria, they have a defined function on the human organism
to improve immune function, prevent specific disease, support recovery from
specific diseases, control physical and psychic complaints, and slow down the
ageing process.
3) Probiotics, Prebiotics.
4) a) Fibre, Phenolic compounds, NDOs, Phytosterol, Lutein, Lycopene
b) Prebiotics, Probiotics, n-3 fatty acids and polyphenols
c) Any three of the following: Polyphenols, Fructans, n-3 Fatty Acids, Phytate,
Carotenoids, Polyphenols, non-glycerides
5) The food can be made functional by the following ways:
elimination of components e.g. toxins or allergenic proteins; increasing the
concentration of a natural component e.g. fortification; addition of components
with beneficial effects e.g. non-vitamin antioxidants; addition of beneficial microbes
e.g. some yeasts, bacteria; replacement of a component, usually a macro-nutrient
e.g. fat replaced with modified or emulsified carbohydrates; enhancement
of biavailability of components.
6) The major components of non-glyceride fraction in edible oils and fats are
sterols, terpene alcohols, tocopherols, hydrocarbons, long chain alcohols including
waxes, carotenoid pigments, sulfur- and nitrogen -containing flavour compounds.

Check Your Progress Exercise 2

1) a) Probiotic is a live microbial feed supplement which beneficially affects the
host animal by improving its intestinal microbial balance for example,
bifidobacteria and lactobacilli.
b) Prebiotic is a non-digestible food ingredients that beneficially affect
the host by selectively stimulating the growth and/or activity of one or a
limited number of bacteria in the colon for example, bifidobacteria and
lactobacilli.
c) NDOs is oligomeric carbohydrates, which are resistant to hydrolysis by
intestinal digestive enzymes but can be metabolized by colonic bacteria for
example, FOS, Soybean Oligosaccharides.
Advance Nutrition 2) Probiotics are rnicroorganis~nswhich promote healthy gut rnicroflora while pre
biotics are non-digestible food components which are fermented in the colon and
alter the composition of rnicroflora there. Snccharomyces boulardi is a probiotic
while non-digestible oligosaccharides are examples of prebiotics.
3) Colonization of the gut appears to be a prerequisite 'for the probiotic effect.
Irnpoi-tant relevant features may be resjstance to low pH and bile acids and
ability to adhere to intestinal epithelium.
4) The physiological effects of probiotics are: (any two of the following)
r In animals, probiotics may control tumour production; in humans, probiotics
c m reduce incidence of intestinal infections; 50% reduction in AAD observed
in patients given Saccharomyces boulardii; significant reduction in diarhoea
incidence among children aged 4-45 months after administration of L.casei
GG; and an antifungal effect.
The beneficial effects of prebiotics are as: (any two of the following)
r control of transit time and motility; regulation of epithelial cell proliferation;
influence on nutlient bioavailability; modulation of immune f~lnclion;and
modulation of endocrine function.
5) The benefits attributed to intake of NDOS include: increases solubilization of
minerals and hence calcium/magnesium absorption and balance, decreases fasting
blood glucose and hepatic glucose production, a hypotriglyceridenlic effect and
decreased lipogenesis and decrease in uremia.
6) Look up sub-section 11.5.3 under fructans and answer on your own.
Check Your Progress Exercises 3
1) Polyphenols contain an OH group attached to the benzene ring. They arise from
phenylalanine or its precursor shikimic acid.
2) Look up sub-section 11.6.2 and answer on your own.
3) a) Influence of polyphenols on minerals include:
Reduced protein digestibility md a subsequent increase in the Pdecal ilitrogen.
Decreased protein digestibility may be due to the foiinalion of protein
tannin complex, as well as, inhibition of proteolytic enzyines like trypsin by
polyphenols.
Inhibition of amylolytic enzymes and subsequent reduction o.I' dietary
carbohydrate hydrolysis can decrease the postprandial glycemic response.
Both soluble polyphenols and condensed tannins have been shown to
decrease the apparent digestibility of lipids, lipase activity and increase
faecal fat excretion.
b) Polypheilols can form complexes with metal 'cations through their carboxylic
or hydroxylic groups, and thus interfere with the intestinal absorption of
minerals like iron, copper, sodium and aluminunl.
4) Look up sub-section 11.6.4 and answer on your own.
5) The antioxidant action of polyphenols can be attributed to the following
characteristics:
- act as terminators of free radicals by rapid donation of H atoms,
- chelate metal ions like iron and copper,
- act as terminators of propagation,
- quench reactive oxygen species and reactive N2 species,
- inhibit lipoxygenase and cyclooxygenase enzyines, and
- inhibit damage by haemproteildperoxide mixtures.
6) i) - c) Food Components other
than Essential Nutrients
ii) - e)
iii> - a)
iv) - b)
v)- d)
Check Your Progress Exercise 4
1) Phytoestrogens are oestrogenic compounds found in plants. They exert oestrogenic
effects on the central nervous system, induce estrus and stimulate growth of the
genital tract of female animals. PE can be divided into 3 main classes: isoflavones
eg: daidezin and genistein, coumestans such as clover and alfalfa sprouts and
lignans eg: secoisolarciresinol and matairesinol.
2) Flax seed, unhulled soyabean, dried sea weed, husk and bran of cereals and
legumes, fruits and vegetables.
3) a) Toxic effects of saponins are:
- A well-known toxic effect is 'erythrocyte lysis' since they interact with the
cholesterol in the erythrocyte membrane
- In mammals, intravenous administration causes local inflammation
- In large doses, it results in death due to massive release of erthyrocyte
debris
- A reduction in oxygen-canying capacity of blood
- It can lyse other cells
- High saponin intake results in decreased weight gain.
b) These are growth inhibition in animals and diarrhoea, nausea, bloating and
vomiting in case of human beings. When injected lectins agglutinate RBCs,
haemolysis and death occurs in extreme cases. Toxic effect of lectins also
relates to their binding with specific receptor sites on intestinal mucosal
cells resulting in lesions, disruption and abnormal development of microviIli.
4) Protective benefits are particularly in relation to carcinogenesis, particularly
against colon cancers; lowering blood glucose and lipids; binding amylase and
reducing starch digestion; lowering plasma cholesterol and triglycerides;
hypoglycemic effects; reduction of blood lipids and reduction in cancer risk.