HARAMAYA UNIVERSITY

SCHOOL OF GRADUATE STUDIES

REVIEW ON THE BIOAVAILABILITY OF PHYTOCHEMICALS AND THEIR ROLE IN

NUTRITION AND HEALTH

Graduate Seminar of Current Topics to the Field of Specialization (FSTC

531, credit hr.1)

By: Habtamu Kefale

Id no: K/ SGS/0278/05

Faculty: Institute of Technology

School: Food Science, Postharvest Technology and Process Engineering

Program: Food Science and Technology

July 2015
Haramaya University, Haramaya
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LIST OF ABBREVIATIONS

ADME Absorption distribution metabolism and excretion

CaCo-2 Colonic adeno Carcinoma
FDA Food and Drug Administration
LDL Low Density Lipoprotein
ORAC Oxygen Radical Absorbance Capacity
UTI Urinary Tract Infection
UGT Uranosyl Gluco Transferase
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TABLE OF CONTENTS

LIST OF ABBREVIATIONS ..................................................................................................... II

LISTS OF TABLES ..................................................................................................................... iv

1. INTRODUCTION .................................................................................................................... 1

2. LITERATUR REVIEW ........................................................................................................... 3

2.1. BIOAVAILABILITY OF PHYTOCHEMICALS ............................................................................... 3

2.1.1. Digestion principles of human gastrointestinal tract .................................................... 4
2.1.1.1. Digestion of Phenolics ........................................................................................... 7
2.1.1.2. Digestion of Carotenoids ....................................................................................... 8
2.1.1.3. Digestion of Organic acids .................................................................................... 8
2.1.2. Absorption of phytochemicals ...................................................................................... 8
2.1.2.1. Absorption of Phenolics....................................................................................... 10
2.1.2.2. Absorption of Carotenoids ................................................................................... 11
2.1.2.3. Absorption of Organic acids ................................................................................ 11
2.2. ROLE OF PHYTOCHEMICALS IN NUTRITION AND HEALTH...................................................... 11
2.2.1. Biological activities of phytochemicals ...................................................................... 11
2.2.2. Health Benefits of Phytochemicals ............................................................................. 13
2.2.2.1. Antioxidant Activity ......................................................................................... 14
2.2.2.2. Cardiovascular Diseases ...................................................................................... 15
2.2.2.3. Glycemic Control and Diabetes ........................................................................... 16
2.2.2.4. Aging ................................................................................................................... 17
2.2.2.5. Vision ................................................................................................................... 18
2.2.2.6. Anticancer activity ............................................................................................... 18
2.2.2.7. Urinary tract health .............................................................................................. 19

3. CONCLUSIONS AND RECOMMONDATIONS ............................................................... 21

3.1. CONCLUSIONS ..................................................................................................................... 21

3.2. RECOMMENDATIONS ........................................................................................................... 22

4. REFERENCES ........................................................................................................................ 23
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LISTS OF TABLES

1. Summary of digestion, characteristics of infant, adult and elderly, female and male
2. Bioactive phytochemicals in medicinal plants
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Abstract

Since global recognition of the dietary guidelines that include increased consumption of Plant-
based foods for the prevention of chronic diseases and maintaining good health, there have been
considerable interests in the biologically active compounds that are present in plant foods. These
compounds have been referred to as ‘Phytochemicals; ‘Phytonutrients’, ‘Nutraceuticals’ and
‘Functional ingredients”. They include a multitude of compounds having different chemical
identities, biological activities and mechanisms of action. The scope of ‘Phytochemicals’ has
expanded beyond their initial applications to food to include therapeutics, pharmaceuticals and
cosmeceuticals. Phytochemicals could provide health benefits as: (1) substrate for biochemical
reactions; (2) cofactors of enzymatic reactions; (3) inhibitors of enzymatic reactions; (4)
absorbents/ sequestrants that bind to and eliminate undesirable constituents in the intestine; (5)
ligands that agonize or antagonize cell surface or intracellular receptors; (6) scavengers of
reactive or toxic chemicals; (7) compounds that enhance the absorption and or stability of
essential nutrients; (8) selective growth factors for beneficial gastrointestinal bacteria; (9)
fermentation substrates for beneficial oral, gastric or intestinal bacteria; and (10) selective
inhibitors of deleterious intestinal bacteria. Such phytochemicals include terpenoids, phenolics,
alkaloids and fiber. Research supporting beneficial roles for phytochemicals against cancers,
coronary heart disease, diabetes, high blood pressure, microbial, viral and parasitic infections,
psychotic diseases, spasmodic conditions, ulcers, etc is based on chemical mechanisms using in
vitro and cell culture systems, various disease states in animals and epidemiology of humans.

Keywords: functional foods; nutraceuticals; phytochemicals

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1. INTRODUCTION

Phytochemicals (from the Greek word phyto, meaning plant) are biologically active, naturally
occurring chemical compounds found in plants, which provide health benefits for humans further
than those attributed to macronutrients and micronutrients (Hasler and Blumberg ,1999).

Phytochemicals are compounds obtained from plants that exert particular health effects which
are not necessarily basic nutrients (minerals, vitamins, carbohydrates, proteins or lipids),
medicines or toxins but increasingly accepted as health promoting, maintaining, and repairing
agents in cells, tissues, or the whole human body. In general, the plant chemicals that protect
plant cells from environmental hazards such as pollution, stress, drought, UV exposure and
pathogenic attack are called as phytochemicals (Gibson.et al, 1998; Mathai et al, 2000).

More than 5,000 phytochemicals have been cataloged (American Cancer Society, 2000) and are
classified by protective function, physical characteristics and chemical characteristics (Meagher
and Thomson, 1999) and about 150 phytochemicals have been studied in detail (American
cancer society, 2000).

Phytochemicals accumulate in different parts of the plants, such as in the roots, stems, leaves,
flowers, fruits or seeds.In wide-ranging dietary phytochemicals are found in fruits, vegetables,
legumes, whole grains, nuts, seeds, fungi, herbs and spices (Mathai ,2000). Broccoli, cabbage,
carrots, onions, garlic, whole wheat bread, tomatoes, grapes, cherries, strawberries, raspberries,
beans, legumes, and soy foods are common sources (Moorachian , 2000).

The phytochemicals that are frequently associated with human health are phenolics, carotenoids,
organic acids, and several miscellaneous bioactive compounds such as saponin and sterols. These
compounds are known as secondary plant metabolites and have biological properties such as
antioxidant activity, antimicrobial effect, modulation of detoxification enzymes, stimulation of
the immune system, decrease of platelet aggregation and modulation of hormone metabolism and
anticancer property. It is well-known that plants produce these chemicals to protect themselves,
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but recent researches demonstrate that many phytochemicals can also protect human against
diseases (Narasinga Rao et al, 2003).

Even though various genetic-base studies propose mechanisms and health interventions of
phytochemicals (Noe et al., 2004), many findings are inconclusive and the roles of
phytochemicals in health are poorly understood, which warrant the needs for validation as well
as scientific database on safety issues and mechanisms of the functions.

Therefore, the purpose of this review was to evaluate the bioavailability of phytochemicals and
their role in nutrition and health as well as to give emphasis on those biologically active and
nutrinatious chemicals.

OBJECTIVE
 To review the bioavailability of phytochemicals and their role in nutrition and health
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2. LITERATUR REVIEW

2.1. Bioavailability of phytochemicals

The bioavailability is an overall result of absorption, distribution, metabolism and excretion

(ADME).
 Absorption describes a compound’s capability to pass into the systemic circulation
following oral administration.
 Distribution explains how well a compound reaches the target tissue.
 Metabolism is the rate that a nutrient/compound is eliminated from the systemic
circulation, following its initial absorption.
 Excretion is the rate that a compound is excreted from the systemic circulation and
ultimately the body. Thus, bioavailability is determined by the combination of the rates of
all these factors.

The bioavailability of a compound can therefore be defined as the amount of nutrient/compound

that reaches the blood circulation system and ultimately the target tissue. Therefore, only the
unbound fraction of a compound will reach the target tissue to be able to interact with the
molecular target (Balani et al., 2005; Van de Waterbeemd et al., 2003).

According to U.S. Food and Drug Administration (FDA), the definition of bioavailability is “the
rate and extent to which the active ingredient or active moiety is absorbed from a drug product
and available at the site of action” (Serrano, et al, 2009). As the definition implies, the rate of
absorption and the availability at the site of action is of utmost importance for a bioactive to be
effective within biological systems and thus be “bioavailable”. Therefore, it is imperative to
determine the amount of a specific nutrient or bioactive compound in a food or dietary
supplement as well as its bioavailability.
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2.1.1. Digestion principles of human gastrointestinal tract

It is known that major phytochemicals are located inside vacuoles of plant cells; and several
phenolics form complexes with fibres in the cell wall. These natural existences make the
phytochemicals poorly accessed by enzymes or hardly released out from the plant matrices
during digestion. Most cell wall materials are indigestible by human enzymic systems.
Therefore, digestibility of the phytochemicals is of great interest, in particular, to reveal how the
phytochemicals can affect human health and fight or prevent diseases.

The digestion compartments in human consist of mouth, gastric, small intestine, and colon as
shown in (Figure 1). Each has slightly different digestion performances depending on age and
gender as listed in Table 1.

In the gastrointestinal tract, net nutriome is released as a result of orchestrated secretions,

enzymic activities, and physical-mechanical actions of peristaltic movements. The nutriomes will
diffuse out from the food particles to chyme solutions. The levels of nutriome in this stage are
called availability or accessibility of the components. However, bio-chemical degradations of the
molecules can take place. Hence, digestibility will also provide metabolites derivatives.

Architecture and material of the plant tissues is generally unfavourable for activities of enzymic
system in the human gastrointestinal tract. As a consequence, limited cell contents of the
ingested food materials are released into chyme solution in the gastrointestinal tract. Natural
pores and plasmodesmatas may not play predominant roles in diffusion of the nutriome.
Nevertheless, according to Stolle-Smits et al. (2004), natural matrices of tomato, mango, apple,
and kiwi undergo galactan solubilisation during ripening stage; thus the release of nutriome can
be altered. However, processing and chemical compositions of the food matrices themselves may
change physicochemical environments of the chyme for nutriome mass transfer.

The most recent finding indicates that ingested foods are necessarily designed such that the
diffusion of the nutriome favours nutriome absorption by epithelial cells; even for phenolics, it
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requires lipid-complex called phytosome (Kidd &Head, 2005) to penetrate gut lining and to enter
the circulatory system.

Fig.1. Principles of human digestion system, Johnson (2001)

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Table 1. Summary of digestion characteristics of infant, adult and elderly, female and male
Digestion Infants Adults Elderly Female Male
Sites
Improper Chewing, Improper Chewing Chewing
chewing,imperfect complete chewing
Mouth salivary enzymes salivaryenzymes may be
with
incomplete
salivary
enzymes
Acid & pepsin Acid & Acid & Acid & Acid &
Gastric Digestion pepsin pepsin pepsin pepsin
digestion digestion digestion digestion

Immature tissues Complete Complete hormonal Hormonal

Small tissues and Tissues related related
intestine enzymes and digestive digestive
enzymes secretion, secretion,
undisturbed disturbed by
byreproductive reproductive
organ cycles organ cycles
Predominant Lossing Lossing Lossing Lossing
Large Bifido bacteria Bifido Bifido Bifido Bifido
intestine bacteria bacteria Bacteria Bacteria

Source: Epriliati (2008)

Cell wall materials significantly modulate digestion of plant foods; Nunan et al. (1998) stated
that in grape berry during its development of the berry fruits the Na2CO3-soluble fraction
increases before veraison but decreases as the berries softened. It implies that the Na2CO3
soluble fraction is the cell wall component which is responsible for firmness and strength.
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Epriliati (2008) observed that ripe mango, tomato, and papaya behave differently when Na2CO3
is added into in vitro digestion model mimicking small intestine where not all aggregated boli
from human in vivo chewing can be broken down. These imply that consumption of fresh and
processed various fruits, rich in pectin, can yield a wide range of phytochemical bioavailability
depending on their cell wall material compositions.

2.1.1.1. Digestion of Phenolics

There are two main routes for digestion of dietary phenolics; i.e. digestion along the
gastrointestinal tract and digestion inside the enterocytes. This can happen because hydrolase
enzymes, i.e. lactase phlorizin hydrolase are available in intestinal lumen, brush border, and
enterocyte (Williamson, 2004).

Metabolisms that take place along the gastrointestinal tract are mainly aiming at deglycosilation
of glycoside form of dietary phenolics. This deglycosilation is also carried out by microbiota in
the colon. Inside the enterocyte, dietary phenolic glucuronidation of the aglycone form are
catalyzed by UGT. Meanwhile, the glycone forms are hydrolyzed and conjugated. The
conjugated forms from both glycone and aglycone dietary phenolics are either effluxed into
intestinal lumen or translocated into the portal blood vessel. The circulated conjugates of dietary
phenolics in plasma can be absorbed by liver and hepatocytes will metabolize them further.

Phenolics stability is strongly affected by pH as studied by Ginjom (2009). For example, syringic
and p-coumaric acids are stable at pH 2-9 for 24 hours. Generally, pH higher than 7.4 is
unfavourable for phenolics and the effects of high pH are worsened by lengthy exposures. The
number of -OH groups in benzene ring of simple phenolics can also be critical clues for phenolic
stability.

Phenolics in tomato products are released into digest solutions more during in vitro gastric
digestion than during pancreatic digestion and the highest release is from tomato juice(Epriliati,
2008 ). The main phenolics in tomato are caffeic, catechin, rutin, chlorogenic acid, and coumaric
acids. More phenolics are obtained from tomato juice than those from dried and fresh tomato
indicating the natural barrier of cell wall has been eliminated.
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Recently, phenolics in gastrointestinal tract markedly behave in a similar way to that of

carotenoids incorporated in chylomicrons, thus, all emulsified phytochemical compounds are
called phytosome (Kidd & Head, 2005). Therefore, the presence of pancreatic juices and bile
extract improve phenolics release during consecutive gastric-intestinal digestions of mango.

2.1.1.2. Digestion of Carotenoids

About 50% of extractable carotenoids dominated by lycopene and carotene in tomato, mango,
and papaya products are released to digest solution in a non-lipidic digestion model (Epriliati,
2008). The release of carotenoids increases significantly in intestinal digestion where bile
extract and pancreatic secretions exist. Consecutive gastrointestinal digestions do not help with
higher release of carotenoids. This is more likely due to insufficient emulsifier-water ratios to
provide emulsification of carotenoids which are fat soluble. It is concluded that mango, tomato,
and papaya carotenoids are released better in intestinal digestion where the model is without
addition of oil (Epriliati, 2008).

2.1.1.3. Digestion of Organic acids

The digestion of organic acids is better when the digestion is takes place inside the gastric and
pancreatic digestion (in vivo) but not by taking samples and underging digestion through
chemicals (in vitro). For example pectin content in tomato hinders organic acid release thus the
total organic acids in in vitro gastric digest solution is lower than that of consecutive gastric-
pancreatic digestion. This is evidenced by the changes in pH from highly acidic gastric pH to
higher small intestinal pH (~6), that causes disaggregation of boli during in vitro digestion. For
all types of mango samples, organic acid including ascorbic acid (Vitamin C) is released better
during gastric digestion. Apparently, the pectinous materials in mango do not trap organic acids
(Epriliati, 2008).

2.1.2. Absorption of phytochemicals

Currently, there is no well-established molecular form of absorbed substances in the

gastrointestinal tract, i.e. whether they are absorbed intact or as metabolites. On the other hand, it
is well known that lifestyle, behavior, diets, and basal metabolism of the subjects are more
important affecting factors than age, gender, body weight, and plasma volume (Manners et al.,
2003) in bioavailability determination. Therefore, standardized experimental conditions
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controlling such critical factors of absorption in vivo and in vitro is a must despite individual
human variability.

The main absorptive tissue is the small intestine. In human 81% of the total intestinal lengths is
by small intestine and 19% is large intestine. The stretched length of jejunum is around 30-78%
of the intestinal lengths (Vermeulen, 2009).

A single enterocyte has microvilli and each microvillus has glycocalyx. Such structure
considerably increases contact surface areas with luminal contents. Each microvillus also
contains a complex structure providing various facilities for uptake/influx and efflux molecules,
signalling ports, cytoplasm, and lipid matrix. The glycocalyx and microvilli are the areas where
the human body depends on for collecting nutriomes but rejecting hazardous compounds
including microbes. Each enterocyte attaches onto adjacent cells through junctions.

Cellular transport from intestinal lumen to portal blood vessel occurs in two ways: paracellular
and transcellular. The paracellular entrances for hazards are tightly controlled by three types of
junction’s tight, adherence and gap. The enterocytes collect compounds that reach apical side.
The compounds then traverse into basolateral side where they end up in capillary vessel for
circulation into the whole human body. Other barrier in intestine is mucus.

Most absorptive tissues comprise of epithelial cells which protect the human body from
hazardous components in ingested foods. The epithelial cells are critical gate for human body.
Due to its critical roles, the epithelial cells along gastrointestinal tract are covered with mucus
secreted by goblet cells making an unstirred water layers so that the coarse particles are not
abrasive towards the epithelial cells.

Epithelial cell membrane is an important part of transport facility. It controls and selectively
takes up molecules required for living or treats hazardous molecules. The transport modes
include passive and active mechanisms. Passive transport is transcellular or paracellular
transports and cynocitosis. Active transports are characterized by the use of protein transporters:
channels/pump, binding protein transport, and formation of vehicles that is mainly emulsion
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system incorporating oil soluble compounds, such as chylomicrons. The transporter is able to
promote transmembrane movement without hydrolyzing ATP (Johnson, 2001). They are
categorized as uniporter (single compound moving down along the electrochemical gradients)
and symporter (two molecules at the same time moving in one direction) or antiporter (two
molecules at the same time moving in opposite directions).

There are two groups of nutriome: water soluble and less polar-solvent soluble. The water
soluble components diffuse out from the food particles into chyme, traverse across the epithelial
lining cells along the brush borders, and enter the portal blood circulation. On the other hand, the
lipid soluble nutriome will be emulsified by bile salts and lipidic components of diets
immediately after diffusing out from the food particles. The emulsion acts as vehicles moving
along the intestinal lumen. Contacting with the epithelial brush border and unstirred water layer
on the top of the epithelial lining cells, rearrangement of vehicle emulsion take place which
eventually releases the lipid soluble compounds into the cells. These compounds then traverse
across the epithelium cells and end up in the lymph circulation.

2.1.2.1. Absorption of Phenolics

Many studies support evidences that aglycone polyphenols are not only absorbed in the small
intestine but also in the large intestine after microbial digestions. The steps may involve
hydrolysis of sugar moiety by intestinal enzymes. In the human small intestine and stomach,
95% of caffeic acid is absorbed while 62% of its ester form (called chlorogenic acid) is reduced.
All are absorbed intact, except chlorogenic acid which mostly enters the human body from colon.
Proanthocyanidins are pH sensitive, thus it is likely to be broken down in stomach so that they
may be readily absorbable (Ginjom, 2009). PH, temperature presence/absence of alcohol and
resident time are necessarily taken into account in order to avoid underestimation/ overestimation
regarding absorption stability (Manach et al., 2005).

In determination of phytochemical bioavailability, researchers should not limit their detection for
ingested molecular forms only based on reported presence in the diets. It has been proven that at
plasma levels many phytochemicals have been conserved by digestion and by hepatic activity.
The ranks of phytochemical absorption is gallic acid and isoflavones > catechins and flavanones,
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quercetin glucoside > proanthocyanidins, galloylated tea catechins, and anthocyanins (Moreno et
al., 2006).

2.1.2.2. Absorption of Carotenoids

Carotenoids of mango, tomato, and papaya in colonic adeno carcinoma (CaCo-2) absorption
model are not detected (Epriliati, 2008) in spite of in vivo data indicates that carotenoid plasma
level increase after consumption of carotenoid-rich foods. Processing altered matrices of ingested
food system and more likely degraded carotenoids which caused variation in bioavailability of
carotenoids. A comparative study of organic and inorganic carrot found that apparently organic
farming practices do not affect bioavailability of carotenoids in carrot consumption (Stracke et
al., 2009).

2.1.2.3. Absorption of Organic acids

Organic acid provides organic anion important for metal binding and counteracting acidosis as
well as preventing chronic diseases (Sabboh et al., 2011). Particular organic acids are apparently
absorbed into plasma. Most organic acids in tomato, mango, and papaya products are absorbed in
in vitro CaCo-2 model but they are not found in the basolateral sides (Epriliati, 2008). On the
other hand, citric acid and oxalic from banana and sweet potato are consistently found to be
absorbed and translocated into basolateral sides in in vitro CaCo-2 model (Sabboh et al., 2011).

2.2. Role of phytochemicals in nutrition and health

2.2.1. Biological activities of phytochemicals

The phytochemicals present in plants are responsible for preventing disease and promoting
health have been studied extensively to establish their efficacy and to understand the underlying
mechanism of their action. Such studies have included identification and isolation of the
chemical components, establishment of their biological potency both by in vitro and in vivo
studies in experimental animals and through epidemiological and clinical-case control studies in
human.
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Study findings suggest that phytochemicals may reduce the risk of coronary heart disease by
preventing the oxidation of low density lipoprotein (LDL) cholesterol, reducing the synthesis or
absorption of cholesterol, synthesis or absorption of cholesterol, normalizing blood pressure and
clotting, and improving arterial elasticity(Mathai.K ,2000 and Abuja, 2003). Phytochemicals may
detoxify substances that cause cancer. They appear to neutralize free radicals, inhibit enzymes
that activate carcinogens, and activate enzymes that detoxify carcinogens. For example,
according to data summarized by Meagher and Thomson, genistein prevents the formation of
new capillaries that are needed for tumor growth and metastasis (Meagher.E and Thomson.C,
1999). The physiologic properties of relatively few phytochemicals are well understood and
more much researches have focused on their possible role in preventing or treating cancer and
heart disease (Mathai K, 2000).

Phytochemicals have also been promoted for the prevention and treatment of diabetes, high
blood pressure, and macular degeneration (American Cancer Society, 2000).
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Table 2. Bioactive phytochemicals in medicinal plants

Classification Main groups of compounds Biological function

NSA Cellulose, hemicellulose, Water holding capacity, delay

(Non-starch polysaccharides.) gums, mucilages, pectins,
lignins
Antibacterial & Terpenoids, alkaloids,
Antifungal phenolics
Antioxidants Polyphenolic compounds,
flavonoids, carotenoids,
tocopherols, ascorbic acid
Anticancer Carotenoids, polyphenols,
curcumine, Flavonoids
Detoxifying Reductive acids, tocopherols,
Agents phenols,indoles, aromatic
isothiocyanates, coumarins,
flavones, carotenoids,
retinoids,cyanates,phytosterols
Other Alkaloids, terpenoids, volatile
flavor compounds, biogenic
amines
in nutrient absorption, binding
toxins and bile acids
Inhibitors of micro-organisms,
reduce the risk of fungal
infection
Oxygen free radical quenching
, inhibition of lipid
peroxidation
Inhibitors of tumor, inhibited
development of lung cancer,
anti-metastatic activity
Inhibitors of procarcinogen
activation, inducers of drug
binding of carcinogens,
inhibitors of tumourogenesis
Neuropharmacological agents,
anti- oxidants, cancer
chemoprevention

Source: Abuja (2003)

2.2.2. Health Benefits of Phytochemicals

Phytochemicals of the bioactive non-nutrient plant compounds in fruit, vegetables, grains, and
other plant foods have been linked to reductions in the risk of major chronic diseases. It is
estimated that more than 5000 phytochemicals have been identified, but a large percentage still
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remain unknown (Shahidi and Naczk , 2004) and need to be identified before their health
benefits are fully understood.
However, more and more convincing evidence suggests that the benefits of phytochemicals in
fruit and vegetables may be even greater than is currently understood because oxidative stress
induced by free radicals is involved in the etiology of a wide range of chronic diseases (Ames
and Gold, 2001).

Cells in humans and other organisms are constantly exposed to a variety of oxidizing agents,
some of which are necessary for life. These agents may be present in air, food, and water, or they
may be produced by metabolic activities within cells. The key factor is to maintain a balance
between oxidants and antioxidants to sustain optimal physiologic conditions in the body.

Overproduction of oxidants can cause an imbalance, leading to oxidative stress, especially in

chronic bacterial, viral, and parasitic infections (Liu and Hotchkiss, 2002). Oxidative stress can
cause oxidative damage to large biomolecules such as proteins, DNA, and lipids, resulting in an
increased risk for cancer and cardiovascular disease (Ames and Gold, 2001). To prevent or slow
down the oxidative stress induced by free radicals, sufficient amounts of antioxidants need to be
consumed. Fruit and vegetables contain a wide variety of antioxidant compounds
(phytochemicals) such as phenolics and carotenoids that may help protect cellular systems from
oxidative damage and lower the risk of chronic diseases.

2.2.2.1. Antioxidant Activity

Numerous studies have evaluated the in vitro antioxidant activity of Vaccinium constituents.
Chlorogenic and caffeic acids inhibited linoleic acid oxidation (Ohnishi et al., 1994). Crude
acidified ethanol extracts from four V. corymbosum cultivars were studied for their antioxidant
activity, but this activity was not directly related to anthocyanin or polyphenol content of the
extracts (Costantino et al., 1992).

Cv. Collins was the most effective inhibitor of both NBT reduction by superoxide anion and
xanthine oxidase, and it had the highest concentration of both anthocyanins and polyphenols.
Velioglu and coworkers (1998) compared the antioxidant activity of methanol extracts from 28
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food and medicinal plants using a β-carotene bleaching assay. V. angustifolium cv. Fandy from
Nova Scotia had greater antioxidant activity than other products studied, with the exception of 50
mg/L α-tocopherol, 200 mg/L BHT, and a few other botanical products such as sea buckthorn
and buckwheat hulls.

The oxygen radical absorbance capacity (ORAC) test is an assay which can assess hydrophilic
antioxidants for their ability to reduce free radicals. Blueberry and bilberry ORAC values were
correlated with anthocyanins (r = 0.77) and total phenols (r = 0.92) (Cao et al., 1998), with
variation among species and cultivars. The researchers concluded that ascorbic acid played only
a minor role in ORAC activity.Oxidation of human low density lipoprotein (LDL) oxidation is
involved with development of atherosclerotic plaques in arteries. Vaccinium phenolic
compounds vary in ability to retard LDL oxidation (Meyer et al., 1998); however, additive
effects were lower than expected.

2.2.2.2. Cardiovascular Diseases

Arterial Relaxation

Cardiovascular disease, as well as the many diseases involved with vascular health, can be
expected to increase in the U.S. population as the proportion of older adults grows. Schramm and
German (1998) have summarized potential benefits of flavonoids in preventing vascular disease,
including: prevention of oxidation and Maillard reactions, inhibition of leukocyte adhesion,
antimicrobial activity that prevents inflammation, and estrogenic effects. Hypertension,
atherosclerosis, and diabetes decrease arterial flexibility, contributing to poor blood flow and
plaque formation.

Capillary Permeability

Anthocyanins protected blood vessels by stabilizing membrane phospholipids and by increasing

production of the acid mucopolysaccharides of the connective ground substance. Anthocyanin
extracts inhibited porcine elastase in vitro (Jonadet et al., 1983). Capillary strength is another
 16

important element in cardiovascular health. Easily damaged or porous capillaries contribute to

electrolyte imbalances, and lead to edema and other dysfunctions.

The commercial V. myrtillus preparation Difrarel containing 20 mg of anthocyanins was

tolerated well by patients with chronic illnesses affecting blood vessels, even though patient
improvement was mixed (Amouretti, 1972) described the progress of 27 patients treated with
Difrarel 20mg, and concluded that the drug was an effective vascular protective agent. A
commercial V. myrtillus extract, known as Myrtocyan®, with 25% anthocyanidins, was effective
in reducing capillary permeability and increasing capillary resistance (Cristoni and Magistretti,
1987).

Platelet Effects

A related cardiovascular tissue involves platelet aggregation where excessive aggregation has
been known to lead to blood vessel blockage. In a small human study, consumption of cranberry
juice (4 glasses per day for 4 days) inhibited aggregation ex vivo (Wilson and Marley, 2001). A
solution of 30 mg/ml bilberry anthocyanins reduced platelet aggregation in vitro, comparing
favorably with aspirin and other drugs (Zaragozá et al., 1985).

2.2.2.3. Glycemic Control and Diabetes

Diabetes is a leading cause of death, and the incidence of Type 2 diabetes is growing in most
developed nations. The leaves and fruit of V. myrtillus are used in Europe to treat many
conditions resulting from diabetes. High levels of serum glucose trigger many adverse
physiological events. Medical complications of diabetes include microangiopathy, cataracts,
blindness due to retinopathy, neuropathy, decreased resistance to infections, and hyperlipidemia.

Capillary Effects

The capillary walls in diabetic patients thicken due to collagen and glycoprotein deposits. The
thickened capillaries are less flexible and more susceptible to blockage and atherosclerosis.
Based upon this series of experiments, the French scientists recommended that diabetic patients
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be given 500 mg V. myrtillus anthocyanins (VMA) daily, split in 2 doses, for a period of at least
several months (Boniface et al., 1986).

Glycemia

Flavonoids interfere with starch enzymes and therefore affect post-prandial blood glucose levels
because glucose is released gradually in the small intestine. An in vitro study indicated that fiber
affects glucose levels by three mechanisms: increased intestinal content viscosity that slows
diffusion to the brush border, entrapment of glucose, and physical separation of enzyme and
substrate (Ou et al., 2001). Flavonoids inhibit aldose reductase, which converts glucose to
sorbitol (Varma, 1986). This conversion is associated with many of the adverse outcomes
associated with diabetes.

2.2.2.4. Aging

Oxidative Stress

As people aged, balance, coordination, short-term memory, information retrieval, and other brain
tasks are hindered. A lifetime of oxidative stress may induce damages to lipids and proteins that
lead to cellular damage. Dementia results from impaired vascular supply, both chronic and acute,
and from the formation of amyloid plaques associated with Alzheimer’s disease. An animal
study recently provided some evidence that dietary antioxidants can protect the brain from
oxygen-induced damage (Joseph et al., 1998). Differences in polyphenolic composition were
thought to be responsible for varying effects of rabbit eye (cv. Tif) and lowbush blueberry
extracts fed to rats for 8 weeks (Youdim et al., 2000).

Hormonal Influences

The thyroid hormones thyroxine (T4) and triidothyronine (T3) regulate temperature and
metabolism. In rats given intraperitoneal injections of bilberry anthocyanins (200 mg/kg/day) for
5 days, significantly more T3 was found in their brains than rats given only the solvent (26%
 18

alcohol) (Saija et al., 1990). T3 enters the brain via specific transport in the capillaries; therefore
anthocyanins may mediate T3 transport at the capillary level.

2.2.2.5. Vision

Bilberry extract is prescribed in Europe for eyesight, particularly night vision. This health benefit
is the primary reason for the product’s popularity in Japan and Korea, where it is used to relieve
computer-induced eyestrain (Kalt and Dufour, 1997). This benefit may be due, in part, to the
small amounts of carotenoids present in the fruit. However, a double-blind, placebo controlled
study demonstrated that oral doses of anthocyanins are important for regeneration of visual
purple. Bilberry extracts appear to benefit vision in several ways: improved night vision by
enhanced regeneration of retinal pigments, increased circulation within the capillaries of the
retina, inhibition of Maillard reactions in the lens to reduce cataract formation, and protection
from ultraviolet light.

In a study of 20-25-year-olds with good vision, the group (N = 30) receiving bilberry extract had
slightly, but significantly, improved adaption related to night vision compared to the placebo
group (Jackson et al., 1997). Inhibition of aldose reductase by Vaccinium flavonoids should aid
in minimizing lens damage in diabetics since free sugars form Maillard-type products that
contribute to lens clouding.

2.2.2.6. Anticancer activity

Consumption of fruit is linked to reduced risks for several cancers. Research involving
Vaccinium species has focused on in vitro and animal studies. Spray dried V. angustifolium juice
was fractionated for several in vitro assays (Smith et al., 2000). A crude 70% acetone extract
induced quinine oxidase, suggesting potential benefits in inhibiting carcinogenesis.
 19

2.2.2.7. Urinary tract health

Bilberries, blueberries, and cranberries have been used in several countries to treat urinary and
digestive tract infections. In the 1960s, several studies examined the effects of anthocyanins on
pathogenic bacteria.

Urinary tract infections

According to the National Institute of Diabetes and Digestive and Kidney Diseases (2001),
urinary tract infections (UTI) resulted in over 8.5 million doctor visits in 1997. These infections
are painful and can lead to serious infections of other organs, usually the kidneys. Interstitial
cystitis afflicts about 500,000 Americans annually; 90% of these sufferers are women. As much
as half of the older women in long-term care facilities or hospitals may suffer from UTIs
(Monane, et al.1995). Prevention of these infections can significantly reduce medical expenses
and improve quality of life for persons prone to recurrent infections. UTIs develop when
bacteria, and occasionally yeasts and fungi, are introduced into the normally sterile urinary tract,
which consists of the bladder and ureters.

Microbes may enter the tract in a number of ways, primarily from sexual intercourse, poor
hygiene, and catheters. Painful and frequent urination, burning, and fever result when the
bacteria multiply. One widely accepted concept is the use of cranberry juice cocktail to prevent
or treat UTIs. Although the earliest history of this practice is not known, many people assume
that Native Americans discovered the beneficial properties of cranberries (V. macrocarpon)
before the arrival of European colonists. Blueberries and cranberries appear to have unique
phytochemical mixtures that aid in reduction of urinary tract pathogens. Mechanisms for
Reducing UTI Bacteria can be killed in a number of ways: extremes of temperature, radiation,
electrical currents, extremes of pH, and numerous antimicrobial compounds.
 20

Urine acidification

The first theory proposed to explain why drinking cranberry juice seemed to help some patients
was based on the idea that the naturally tart cranberry juice contained acids that lowered the pH
of urine. Normal urine pH is close to neutral (7.0); a pH of 5.5 or lower is necessary to retard
development of UTIs.
The researchers suggested that cranberry juice aided the drug methenamine by keeping urine pH
sufficiently low for the drug to have enhanced antibacterial activity. They concluded that
cranberry juice was a helpful, well-tolerated, and inexpensive adjunct to traditional antibiotic
therapy.
.
 21

3. CONCLUSIONS AND RECOMMONDATIONS

3.1. Conclusions

From the overall review it can be concluded that:

 Phytochemicals are not basic nutrients , medicines, or toxins rather they are biologically
active compounds of plants,
 Digestion is a crucial factor for determination of bioavailabilty of phytochemicals and
how they prevent diseases.

 Phytochemical bioavailability is strongly dependent on cell wall compositions of the food

matrices they originate from, structural chemistry of the phytochemicals.

 Phytochemicals are secondary metabolites and they can prevent diseases like cancer,
urinary problems, used as antioxidant activity, anti microbial activity.

 It is also concluded that availability of phytochemicals depends on digestion site, fruit

type, history of processing, as well as the age of human beings.
 22

3.2. Recommendations

Based on above review the following points are recommended

 Since phytochemicals are not food but components in food, to obtain them consuming
fruits,leafs, steams or seeds of plants based on given scientific evidence is advisable.

 Researchers should undergo further study on the availability of phytochemicals especially

on fruits and vegetables to alleviate chronic, diabetic, vision, aging, cardiovascular
problems.

 Since now a day’s diabetes is a fast growing disease and leading cause of death in the
world, further study is needed on the fruit of variety myritillus.

 Moreover, it is recommended that further study is needed on the health benefits of

carotenoids, flavonids of plant phytochemicals.
 23

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