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CHAPTER - I
INTRODUCTION
TABLE OF CONTENT
HNGU Ph.D Thesis
CHAPTER: 1
Chapter 1 INTRODUCTION……………………………………………….. 1-22
1 Introduction…………………………………………………………….... 1
1.1 Herbal Drugs…………..……………………………………….... 1
1.2 Allergy………………………………………………………….… 2-9
1.2.1 Types of allergy…………………………………………... 3
1.2.2 Treatment of allergy………………………………………. 5
1.2.3 Ayurveda and allergy treatment…………………………... 6
1.2.4 Traditionally used anti-allergic plant……………………... 6
1.2.5 Ayurvedic formulations used in treatment of allergy…….. 6
1.2.6 Experimental models for screening of antiallergic agent…. 7
1.3 Antioxidants…………………………………………………....... 10-12
1.3.1 Models for screening of antioxidant activity…………….. 11
1.4 References………………………………………………………... 13-22
CHAPTER: 1 INTRODUCTION
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1. INTRODUCTION
1.1 Herbal Drugs
Herbal medicines consist of herbs, herbal materials, herbal preparations and finished
herbal products which contain as active ingredients, parts of plants, or other plant
materials, or combinations defined by World Health Organization (WHO).1 Herbs used
as medicinal purposes for many centuries. Now a day, herbal medicines are used globally
as home remedies in treatment of various ailments. People rely mainly on herbal drugs to
meet their primary health care needs in some developing countries. Herbal medicines are
also gaining popularity as alternative and complementary therapies in many industrialized
and developed countries. There has been an increase in scientific studies on herbal
medicines, due to insufficient availability of reliable data still today.
Herbal medicines are an integral part of the development of modern civilization. Most of
the medicinal uses of plants have been developed through observations by trial and error.
Tribes added the medicinal herbs in their regimen of curing various diseases, on the bases
of herbals knowledge observed in routine life. Collection of systematic information on
herbs, ultimately lead to develop well-defined monographs and herbal pharmacopoeias in
different countries.
The WHO estimates 80% of the world population (4 billion) presently consume herbal
medicine for some aspect of primary health care. Herbal medicine is the major
component in all indigenous traditional medicine and a common element in ayurvedic,
homeopathic, naturopathic, traditional, and native. WHO notes that out of 119 plant-
derived pharmaceutical medicines, about 74% are used in modern medicine in ways that
correlated directly with their traditional uses as plant medicines by human beings.
Currently, major pharmaceutical companies are conducting extensive research on plant
materials for their potential medicinal value.
Substances derived from the plants remain the basis for a large proportion of the
commercial medications used today for the treatment of various diseases.2 For example,
ephedra is an herb used in traditional Chinese medicine for more than 2000 years to treat
asthma and other respiratory problems. Ephedrine derived from it is used in the
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HNGU Ph.D Thesis
commercial pharmaceutical preparations for the relief of asthma symptoms. Foxglove
(digitalis) plant used since 1775; presently the powdered leaf of it or active constituents
(digoxin) is known as the cardio tonic.
Traditional medicine is the sum total of the knowledge, skills and practices based on the
theories, beliefs and experiences which are indigenous to different cultures, whether
explicable or not, used in the maintenance of health, as well as in the prevention,
diagnosis, improvement or treatment of physical and mental illnesses. Traditional
medicine has been developed based on history and culture of the countries or regions.
The term “traditional medicine” covers a wide variety of therapies and practices which
vary greatly from country to country and from region to region. Traditional,
complementary, or alternative medicine has many positive features. Traditional medicine
and its practitioners play an important role in treating chronic illnesses, and improving
the quality of life of those suffering from minor illness or from certain incurable
diseases.3
Traditional medicine are facing many challenges and difficulties like; lack of legal
recognition, appropriate law and regulations, knowledge of preservation and protection,
appropriate control mechanism and approaches to control safety, efficacy and quality,
adequate support to the research resources, training, education programmes and licensing
practice for qualified practice of traditional medicine or medicinal plants, practice and
practitioner. 4
It is estimated that at least 25% of all modern medicines are derived, either directly or
indirectly, from medicinal plants, primarily through the application of modern technology
to traditional knowledge. In the case of certain classes of pharmaceuticals, such as
antitumor and antimicrobial medicines, this percentage may be 60%. In developing
countries, 70%–95% of population relies on traditional medicines for primary care.5
1.2 Allergy
About 30-40% of world population is affected by allergic conditions.6 The first allergic
history was reported in the year 2641 BC after wasp sting.7 In the middle age rose fever
with hay fever like symptoms was found. The first clinical description of hay fever was
given by John Bostock8. The term "allergy" was originally introduced by von Pirquet9 in
CHAPTER: 1 INTRODUCTION
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HNGU Ph.D Thesis
1906, meaning "changed reactivity.10 World allergy organization (WAO) define, allergy
is a hypersensitivity reaction initiated by immunological mechanisms. Allergy can be
antibody or cell mediated. In the majority of cases the antibody responsible for an allergic
reaction belongs to the IgE isotype and these individuals may be referred to as suffering
from an IgE-mediated allergy. In non-IgE-mediated allergy the antibody can belong to
the IgG isotype, eg, anaphylaxis due to immune complexes containing dextran. Both IgE
and IgG antibodies are found in allergic bronchial pulmonary aspergillosis (ABPA).
Allergic contact dermatitis is mediated by lymphocytes.
Allergens are antigens which cause allergy. Most allergens reacting with IgE and IgG
antibody are proteins, often with carbohydrate side chains, pure carbohydrates, low
molecular weight chemicals, eg, isocyanates and anhydrides acting as haptens. In allergic
contact dermatitis, the allergens are low molecular weight chemicals reacting with cells
eg, chromium, nickel and formaldehyde.11
1.2.1 Types of allergy12,13,14,15
Allergy is a reaction between immune system and a foreign body or an allergen. The
body’s immune system or the defense mechanism works overtime fighting these
allergens hence, it is also called as hypersensitivity. Steps involve in allergy are i)
appearance of antigen, ii) synthesis of antibodies that is specific to antigen and iii)
antigen –antibody reaction. Allergy is humoral or cell mediated. It can be caused due to
medicines, food, insect, pollen, insect stings, and mold spores. It causes symptoms like
sneezing, runny nose, itching rashes, urticaria, swelling, asthma. Coomb and Gell 16
described four types of hypersensitivity; e.g. type-I or anaphylaxis, type-II or Cytotoxic,
type III or toxic complex reaction and type IV or cell mediated.
Type I Hypersensitivity (Atopic allergy): It is characterized by an allergic reaction that
occurs immediately following contact with the antigen, referred to as the allergen. Atopic
allergy is mediated by IgE, antibodies which are bound to the mast cells. When allergens
cross-link the IgE molecules degranulation is induced and mediators such as histamine
and other vasoactive amines are released and produce the allergic reactions. Total and
allergen specific circulating IgE antibodies can be measured in serum. Common
environmental allergens inducing IgE-mediated allergy are cat, dog and horse epithelium,
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HNGU Ph.D Thesis
tree, grass, weed pollen, house dust mites and molds. Food allergens are also a common
cause of allergy, even though food allergy is more frequently seen in children than in
adults. In recent years "allergy" only has become synonymous with Type I.
The term atopy was coined in 1923 by Coca and Cooke17 to describe clinical allergy of an
inherited nature. Nowadays an atopic constitution implies a hereditary tendency to
produce IgE to common environmental allergens and an increased risk of developing
asthma, rhino-conjunctivitis (hay fever), urticaria and atopic eczema. Atopic patients also
have an increased tendency to be allergic to food substances, which can result in various
symptoms, including described above and others of the gastrointestinal tract.
Type II Cytotoxic type: In these type of immune reactions the antibodies are free in the
serum while antigens are bound to the surface of certain cells or a component of the cell
membrane. Often the substances concerned are small molecules, haptens that are fixed to
the cell surface, thus resulting in the formation of the antigen. When the antibodies (of
IgG or IgM type) react with the antigen, complement is activated, the cell is damaged and
lysis may occur. A complement induced accumulation of granulocytes, together with
release of anaphylatoxins and histamine, can lead to further damage. Drug reactions,
transfusion reactions to erythrocytes, hemolytic disease of the newborn and autoimmune
hemolytic anaemia are some of the examples of type II hypersensitivity. The pathology
depends on the molecules and tissues targeted by the antibodies.
Type III Toxic complex reaction: Precipitins, the free circulating antibodies belong to
the IgG class, cause these reactions.18 The complement system is activated when these
antibodies react with locally introduced antigens to produce an antigen-antibody
complex. This activation of complement, together with complement induced granulocyte
accumulation and release of histamine via anaphylatoxins, results in tissue damage, an
Arthur’s reaction. Allergic alveolitis is an example of such an immune complex reaction:
inhalation of antigens such as molds, plant or animal antigens results in tissue changes,
giving rise to symptoms such as coughing, dyspnoea, fever and cyanosis. The tissue
damage may be irreversible in persistent allergic condition. Persistent infections and
autoimmunity could cause other categories of immune-complex diseases. Another
example of this type reaction is serum sickness, antigen introduction, in the form of a
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HNGU Ph.D Thesis
foreign protein, results in the formation of IgG antibodies. The antigen-antibody complex
formed from circulating antigen and a precipitated antibody is deposited in different
tissues (e.g. lung), resulting in tissue damage. It has been shown that the immune
complex which is formed with an excess of antigen is the most damaging. The symptoms
of serum sickness can partly be of a general nature such as fever, pain in the joints, skin
changes or swelling of the lymph glands, but they can also be localized to organs such as
the heart or the kidneys.
Type IV cell mediated type: This type of hypersensitivity differs from the other types of
reactions in that it is not caused by antibodies, but by lymphocytes cells which are
immunologically specific with receptors for the antigens, tissue antigens or small
molecular substances, fixed to the cell membrane.19 These reactions called delayed
hypersensitivity reactions, which occurs 12-48 hours after exposure to the antigen. It
leads to inflammatory tissue damage and infiltration of cells, which are principally
mononuclear (lymphocytes and macrophages). The inflammatory reaction leads to
irreversible damage with deterioration of the tissue. The classical examples of type IV
hypersensitivity are the positive tuberculin reaction, contact dermatitis (e.g. caused by
nickel or chrome) and rejection of tissues transplanted from other individuals. Histamine
is one of the important mediators of allergy, inflammation and bronchoconstriction,
which were released after degranulation of mast cell by antigen exposure. On activation,
mast cells released immediately the preformed and the de novo synthesized other
mediators proteases, leukotrienes, prostaglandins and cytokines. As a consequence, the
acute reactions such as vasodilation, increased vascular permeability, and
bronchoconstriction will be induced. In addition, allergic responses also trigger the influx
and activation of a variety of inflammatory cells including eosinophils and lymphocytes.
Rapidly released mediators and numerous cytokines produced by mast cells are strongly
believed to induce and sustain these responses, which may contribute to chronic
inflammation. Targeting histamine, either preventing its release from mast cell or
histaminergic receptor antagonist becomes part of therapy in allergic diseases.
1.2.2 Treatment of allergy 20
The avoidance of allergen is first step to controlling allergies. The skin allergy can be
reduced by using soothing creams and wet wrapping. Drugs used to treat allergies;
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i. Drug blocks the activity of mediators which are released during allergic
reactions. e.g. Antihistamines and leukotriene antagonists.
ii. Drugs relax the constricted muscle around the airways of the lungs, or shrink
congested tissue, or reverse the effects of the chemicals released during allergic
reactions .e.g. bronchodilators, decongestants and epinephrine.
iii. Drugs which reduce inflammation. e.g. corticosteroids.
iv. Drug modifies the immune response. e.g. allergen immunotherapy.
antihistamines and leukotriene antagonists
1.2.3 Ayurveda and allergy treatment21,22
In Ayurveda the main causative factor of allergy is from improperly digested food called
Ama. Allergic reaction illnesses (ARIs), drug allergies, or hypersensitivities are
considered Kapha dominated diseases in Ayurveda, e.g. asthma and eczema. In
Allopathic medicine, the treatment of these diseases clusters around the use of steroids,
antihistamines and bronchodilators (beta-adrenergic blockers). The treatment of allergic
reactions in Ayurveda essentially consists of identifying the allergens, avoiding the
exposure to them, using drugs to relieve acute symptoms, improving digestion and
cleaning the intestine of toxic materials in the gut (ama). Emesis is recommended to treat
allergy and asthma because ARI are considered kaphaja disease.
1.2.4 Traditionally used anti-allergic plant 23
Adhatoda vasica, Albezia lebbeck, Allium cepa, Allium Sativa, Alpinia galangal,
Benincasa hispida, Boerhavia diffusa, Calotropis procera, Cedrus deodara, Curcuma
longa, Cyperus rotundus, Datura stramonium, Dolichos biflorus, Elettaria cardamomum,
Emblica officinalis, Fagonia cretica, Galphimia glauca, Glycyrrhiza glabra, Inula
racemosa, Mentha spicata, Mucuna pruriens, Ocimum sanctum, Phyllanthus surinaria,
Picrorhiza kurora, Piper longum, Rhus succedanea, Solanum xanthocarpum, Terminalia
belerica, Tylophora indica, Vitis vinifera, Withania somnifera and Zingiber officinale.
1.2.5 Ayurvedic formulations used in treatment of allergy
Kantaryadikvath, Srmgyadikvath, Vasadikvath, Agastyaharitikiavaleha, Chavanpra-
shavaleha, Chitrakaharitikiavaleha, Vasaavaleha, Vyaghriharitikiavaleha, Draksharishta,
CHAPTER: 1 INTRODUCTION
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HNGU Ph.D Thesis
Vasarishta, Kanakasva, Abhrakabhasma, Apamargaksara, Arkalavana, Khadiradivati and
Lavan-gadivati.
1.2.6 Experimental models for screening of antiallergic agent
A. Acetyl choline or histamine-induced bronchospasm in guinea pig24,25: Asthma
can be experimentally induced in guinea pig by exposing to acetylcholine or
histamine in an aerosol chamber. A guinea pig shows progressive signs of breathing
difficulty leading to convulsions, asphyxia and death. The time until the appearance
of convulsion is called pre-convulsion time (PCD). The end point for PCD is
determined from the time of aerosol exposure to the onset of dyspnoea leading to
the appearance of convulsions. Similarly, PCD is determined with selected extracts.
The protection offered by the extracts can be calculated by the following formula.
Where;
T1 is time for PCD onset (Without treatment) and
T2 is the time for PCD onset. (Treated with extracts)
B. In vitro studies on isolated guinea pig ileum26: Guinea pig ileum is rich with
histamine receptor. Histamine causes contraction of ileum. The contractile response
of ileum to histamine is determined in presence and absence of selected extract.
C. Milk-induced leucocytosis in mice27: The boiled and cooled milk injected in mice
subcutaneously induces leukocytosis, a one of the symptoms of allergic condition.
Difference in total leukocyte count before plant extract administration and after
milk injection indicates the antiallergic potential of plant extracts.
D. Isolated goat tracheal chain28: Isolated adult goat tracheal tissue produce
contraction in presence of histamine, acetyl choline, 5-hydroxytryptamine or
bradykinin. The contractile response of goat tracheal chain to histamine is
determined in presence and absence of selected extract. Percent of maximum
contractile responses is determined in the absence and presence of plant extract.
E. Mast cell stabilization activity29,30: Peritoneal fluid of rats contains mast cell. The
mast cell can be collected from peritoneal fluid. The compound 48/80 causes
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HNGU Ph.D Thesis
degranulation of mast cell and liberates histamine. The selective dye like toluidine
blue stains disrupted mast cells while undisrupted mast cells remain as such.
Percentage protection from degranulation of mast cells can be determined in
presence and absence of plant extracts.
F. Passive paw anaphylaxis in rats31,32: Passive paw anaphylaxis is induced in
Wistar rats egg albumin (s.c.) adsorbed on aluminum hydroxide gel. After
sensitization, blood serum injected in the left hind paw of animals. The contra
lateral paw received unequal volume of saline. Drug treatment is given after
sensitization. Then animals are again challenged in the left hind paw with egg
albumin. The difference in the reading prior to, and after antigen challenge
represented the edema volume and the percent inhibition of volume is calculated by
using the following formula.
Where;
Vt = Mean relative change in paw volume in test group
Vc = Mean relative change in paw volume in control group.
G. Antipruritic activity33: Scratching is induced by administering compound 48/80
subcutaneously in to dorsal surface of the neck of mice. The incidence of scratching
behaviour on the whole body and the site injected with compound 48/80 can be
determined with and without treatment of selected plant extracts.
H. In vitro studies on isolated Rat ileum34: Acetyl choline cause bronchial
contraction, so it exaggerates the allergic condition. Rat’s ileum is rich with the
acetyl choline receptor. The contractile response of ileum to acetyl choline is
determined in presence and absence of selected extract.
Herbs are used in treatments of allergic ailments like hay fever and asthma. These herbs
work to decrease the production of chemicals responsible for allergy. Herbs with natural
antihistamine activity act by reducing formation of histamine.35 The list of some plants
reported antiallergic activity is given in Table1.1.
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HNGU Ph.D Thesis
Table1.1 List of Plants having antiallergic activity
Plant Part Plant Part
Abrus precatorius36 Leaves Gleditisia sinensis37 Fruits
Acanthopanax senticosus38 Root Hemides musindicus39 Root
Ailanthus excelsa 40 stem bark Inula japonica41 Flowers
Albizzia lebbeck42 Bark, flower Lagenaria siceraria43 Leaf
Amaranthus spinosus44 Leaves Lycopus lucidus45 --
Aristolochia Indica46 Root Magnolia officinalis47 Bark
Aristolochia atagala48 Root Mirabilis jalapa49 Root
Artemisia iwayomogi50 -- Momordica dioica51 Fruit
Asystasia gangetica52 Leaf Morus bombycis53 --
Bauhinia variegata54 Bark Myrica esculenta55 stem bark
Bidens pilosa56 -- Myrica sapida57 --
Bunium persicum58 -- Passiflora incarnata59 Leaves
Camellia japonica60 Leaf Perilla frutescens Seed
Carum copticum61 -- Prunella vulgaris62, 63 ---
Cassia alata64 Leaves Pterocarpus marsupium65 Bark
Cassia sophera66 Leaves Punica granatum67 Flower bud
Cecropia glaziovi68 Leaves Rehmannia glutinosa69 --
Centella asiatica70 -- Rumex gmelini Herb
Cichorium intybus71 -- Salvia plebeia72 --
Clerodendron trichotomum73 Leaves Sanseveiria trifasciata74 Leaves
Clitoria ternatea75 Roots Schisandra chinensis76 Fruit
Crocus sativus77 -- Solanum nigrum78 Fruits
Curculigo orchioides79 Rhizome Sphaeranthus indicus80 Whole plant
Cyperus rotundus81 Rhizome Striga orobanchioides82 Whole plant
Dolichos biflorus83 Seed Strychnos potatorum84 Seed
Eclipta alba85 Whole herb Syzygium cumini86 Leaf
Elaeagnus pungens87 Leaf Tephrosia purpurea88 Root
Elephantopus scaber89 Leaves Teucrium japonicum90 --
Ficus bengalensis91 Bark Vitis amurensis92 Fruits
Ficus religiosa93 Leaves Xylopia aethiopica94 Fruit
Gastrodia elata95 Roots Zataria multiflora96 --
German chamomile97 Rhizomes
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HNGU Ph.D Thesis
1.3 Antioxidants
It is widely accepted that antioxidants have protective effect against some pathological
events. There is a worldwide trend toward the use of natural antioxidant.
Reactive oxygen species (ROS) such as singlet oxygen, superoxide anion (O2-) and
hydroxyl (.OH) radical and hydrogen peroxide (H2O2) are often generated as byproducts
of biological reactions or from exogenous factors. These reactive species exert oxidative
damaging effects by reacting with nearly every molecule found in living cells. Such
species are important causative factors in the development of diseases such as diabetes,
stroke, arteriosclerosis, cancer, cardiovascular diseases and the aging process. Human
body has multiple mechanisms especially enzymatic and non-enzymatic antioxidant
systems to protect the cellular molecules against reactive oxygen species (ROS) induced
damage.98 However, the innate defense may not be enough for severe or continued
oxidative stress. Hence, certain amounts of exogenous antioxidants are constantly
required to maintain an adequate level of antioxidants in order to balance the ROS in
human body. When free radicals or oxidants are produced in abundance, cells suffer from
oxidative stress. Fortunately, compounds called antioxidants quickly balance the free
radicals, inhibiting oxidative stress. Antioxidants reduce the effect of dangerous oxidants
by binding together with these harmful molecules, decreasing their destructive power.
Antioxidants can also help to repair damage cells.
The body’s antioxidant supply can be classified into two groups: Primary antioxidants are
made by the body, thus internally provided. This internal antioxidant defence system
includes superoxide dismutase (SOD), catalase and glutathione peroxidase, which are the
first and most powerful line of defence against oxidative stress. Secondary antioxidants
are externally provided from dietary sources, such as vitamins (vitamin A, C and E),
minerals (selenium, zinc, copper and manganese) and other substances, including
polyphenols found in grapes and green tea. These dietary antioxidants contribute to the
antioxidant reserve, yet play a secondary role to the body’s own antioxidants. Choosing
raw fruits and vegetables rather than cooked, provide the highest concentration and best
absorption of antioxidants. Dietary supplements are also available for those that do not
consume enough antioxidant producing foods. Natural antioxidants are structurally
CHAPTER: 1 INTRODUCTION
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HNGU Ph.D Thesis
phenolic or polyphenolic compound occurs in plants. Dietary supplements such as
vitamins A, C and E and flavonoid prevent oxidative damage. Antioxidants are major
five types99.
i. Primary antioxidants: They terminate the free radical chain, acts as electron
donors. e.g. BHT, BHA, Tocopherols.
ii. Oxygen scavengers: They are reacting with oxygen. e.g. Vitamin C
iii. Secondary antioxidants: They decompose lipid hydroperoxidase in to stable end
products.
iv. Enzymic antioxidants: It removes oxygen or highly oxidative species. e.g.
glucose oxidase or super oxide dismutase.
v. Chelating agents: They are synergistic substance which enhances the activity of
phenolic antioxidants. e.g. Citric acid and amino acid.
1.3.1 Models for screening of antioxidant activity
Antioxidant activity cannot be measured directly but rather, by effects of the antioxidant,
in controlling the extent of oxidation strategies have been developed for measuring the
antioxidant activity, as the ability to scavenge free radicals generated in aqueous and
lipophilic phase. The ability to scavenge specific radicals may be targeted as, for
example, hydroxyl radical, super oxide radical or nitric oxide radical.100,101,102
A. DPPH free radical scavenging activity103,104: DPPH (1, 1-diphenyl-2-picryl-
hydrazyl, Deep violet coloured) is a free radical and accepts an electron or
hydrogen radical to become a stable diamagnetic molecule yellow coloured
diphenyl picryl hydrazine. The antioxidants reduce and decolorize DPPH by their
hydrogen donating ability. The reduction capability of DPPH radical by
antioxidants is determined by the decrease in absorbance at 516nm.
B. Reducing power by FeCl3105: Assay of reducing activity is based on the
reduction of Fe3+/ferricyanide complex to the ferrous form in presence of
antioxidants. The Fe2+ is then measured by the formation of Perl's Prussian blue at
700nm using spectrophotometer. Higher absorbance of the reaction mixture
indicates greater reducing power.
CHAPTER: 1 INTRODUCTION
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HNGU Ph.D Thesis
C. Metal chelating activity106: Ferrous ion form red coloured chelate with ferrozine.
The extent of formation of complex is inhibited in presence of other chelating
agent. Resulting, the decreasing intensity of red colour is due to inhibition of
formation of ferrozine-Fe2+complexes. Determination of the colour reduction
measures the chelating activity of drug.
D. Nitric oxide scavenging activity107: Nitric oxide synthases generates NO· radical
in biological tissue. The compound sodium nitroprusside is able to decompose in
aqueous solution at physiological pH (7.2) to produce producing NO· radical. It
reacts with oxygen to produce stable nitrate and nitrite. The amount of nitrite ions
can be estimated by use of Greiss reagent. Scavengers of nitric oxide compete
with oxygen leading to inhibit formation of nitric oxide.
E. Phosphomolybdenum assay108: The antioxidant capacity is determine by
phosphomolybdenum assay based on the reduction of Mo (VI) to Mo (V) by the
antioxidants and subsequent formation of a green phosphate/Mo (V) complex at
acidic pH. The higher absorbance value indicates higher antioxidant activity.
F. Β-carotene linoleic acid method109,110: It based on the principle that linoleic
acid, unsaturated fatty acid, gets oxidized by in presence of oxygen and produced
oxygenated water. It causes oxidation of β-carotene resulting discoloration of β-
carotene. Antioxidant decreases the extent of decolouration of β-carotene. The
extent of antioxidant potential can be measured by inhibition of decolouration of
β-carotene by using spectrophotometer.
G. Hydrogen peroxide scavenging (H2O2) assay111,112: H2O2 rapidly decomposed
into oxygen and water and this may produce hydroxyl radicals cause DNA
damage in biological tissue. The concentration of H2O2 is determined in presence
and absence of antioxidant. As per Haliiwell et al. interaction of iron with free
radical leads to formation of highly active tissue damaging species. The 2-deoxy
ribose reacts with hydroxyl radical in presence of thiobarbituric acid to form pink
chromogen. The concentration of chromogen is measure using spectrophotometer.
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1.4 References
1. http;//www.who.int/medicines/ares/traditional/definations/en.
2. http;//www.holistic –online.com/herbal-med/hol_herb-intro.htm.
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4. Anonyms. Global review: Role of traditional medicine on panel on traditional
medicine; February 12, 2009 ECOSOC New York, USA.
5. Robinson MM, Zhang X. The World medicines situation 2011, Traditional medi-
cines: global situation, issues and challenges, 3rd ed. WHO, Geneva, 2011:2-5.
6. Pawankar R, Canonica GW, Stephen T. Holgate and Richard F. Lockey. WAO
White Book on Allergy, World Allergy Organization, USA, 2011-2012: 12-13.
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8. Jackson M. Allergy: The History of a Modern Malady, Reaktion Books, London,
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9. Cruse JM, Lewis RE. Atlas of Immunology, 3rd Edition, CRC Press, New York,
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10. Platts-Mills T. Johannes Ring. Allergy in Practice. Springer, New York, 2006, 1-3.
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nglish.php.
12. Tripathi KD. Essentials of Medical Pharmacology, 7th ed. New Delhi: Jaypee
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13. Umetsu DT, McIntire JJ, Akbari O, Macaubas C, DeKruyff RH. Asthma: an
epidemic of dysregulated immunity. Nat Immunol 2002; 3:715-20.
14. Roitt I, Brostoff J, Male D. Immunology. 5th ed. Hypersensitivity–Type I. London:
Mosby, 1998:301.
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15. Gandhi TP, Goyal RK. Elements of Pharamcology. 9th edition, BS Shah
Prakashan, 2000, 300-01.
16. Elgert KD. Immunology: understanding the immune system.2nd edition, John
Wiley & Sons, USA, 2009:399-409.
17. Barnes JP, Drazen JM, Rennard SI, Thomson NC. Asthma and COPD: Basic
Mechanisms and Clinical Management Academic Press, New York, 2002:383-384.
18. Calvey N, Williams N. Principles and Practice of Pharmacology for Anaesthetists,
5th edition, John Wiley & Sons, USA, 2009:107.
19. Porth C. Essentials of Pathophysiology: Concepts of Altered Health States, 3rd
Philadeiphia, Lippincott Williams & Wilkins, 2011:351.
20. http://www.worldallergy.org/public/allergic_disease_centre/allergymanagment.php
21. Mishra LC. Allergic Reaction scientific Basis for Ayurvedic Therapies. CRC
Press. New York, 2004, 203-205.
22. Ganapaty S, Chandrashekhar VM, Narsu ML. Evaluation of anti-allergic activity
of gossypin and suramin in mast cell-mediated allergy model. Ind. J Biochem
Biophys. 2010; 47(2):90-5.
23. Mishra LC. Allergic Reaction scientific Basis for Ayurvedic Therapies. CRC
Press. Newyork, 2004, 217.
24. Dhawan K, Kumar S, Sharma A. Antiasthmatic activity of the methanol extract of
leaves of Passiflora incarnata. Phytother Res 2003; 17: 821-22.
25. Mitra SK, Gopumadhavann S, Venkataranganna MV, Anturlikar SD. Anti-
asthmatic and anti-anaphylactic effect of E-721b, A Herbal Formulation. Ind J
Pharmac. 1999; 31: 133-137.
26. Bahekar PC, Shaikh HY, Nigade PB, Ghaisas MM. Anti-histaminic activity of
aqueous extract of leaves of Mimosa pudica Linn. J Pharm Res2007; 6: 134-138.
27. Taur DJ and Patil RY. Effect of Abrus precatorius leaves on milk induced
leukocytosis and eosinophilia in the management of asthma. Asian Pacific J Trop
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