Antioxidant Activity and Preliminary Phytochemical Studies of Various Extracts of Agaricus Bisporus

ANTIOXIDANT ACTIVITY AND PRELIMINARY
PHYTOCHEMICAL STUDIES OF VARIOUS EXTRACTS

OF Agaricus bisporus
A Dissertation submitted to
THE TAMILNADU Dr. M.G.R MEDICAL UNIVERSITY

CHENNAI-600 032
In partial fulfillment of the requirements for the award of degree of
BACHELOR OF PHARMACY
Submitted by
Miss.Divyadarshini.S - 561795221
Mr.Gokul.R - 561795224
Mr.Jerald J Jacob - 561795233
Mr.Mani.D - 561795242
Under the guidance of

Mrs.SRIVIDHYA.V, M.Pharm.,
Professor
Department of Pharmacology
Excel College of Pharmacy

March 2022
CERTIFICATE
This is to certify that the dissertation work entitled “Antioxidant Activity
and Preliminary Phytochemical Studies of Various Extracts of Agaricus
bisporus” submitted by the students bearing REG NO: 561795221,
561795224, 561795233, 561795242 to “The Tamil Nadu Dr. M.G.R.
Medical University”, Chennai, in partial fulfillment for the award of
Bachelor of Pharmacy was evaluated by us during the examination held
on……………………………….
INTERNAL EXAMINER EXTERNAL EXAMINER

CERTIFICATE
This is to certify that the dissertation “Antioxidant Activity and
Preliminary Phytochemical Studies of Various Extracts of Agaricus
bisporus” is a bonafide work done by REG NO: 561795221, 561795224,
561795233, 561795242 at the Department of Pharmacology, Excel College
of Pharmacy, Komarapalayam, in partial fulfillment for the award of
Bachelor of Pharmacy under the guidance and direct supervision of
Mrs.SRIVIDHYA.V, during the academic year 2021-2022.
Mrs.SRIVIDHYA.V., M.Pharm.,
Assistant Professor
CERTIFICATE
This is to certify that the work embodied in this dissertation entitled
“Antioxidant Activity and Preliminary Phytochemical Studies of
Various Extracts of Agaricus bisporus”, submitted to “The Tamil Nadu
Dr.M.G.R. Medical University”, Chennai, in partial fulfillment to the
requirement for the award of degree of Bachelor of Pharmacy, is a
bonafide work carried out by REG NO: 561795221, 561795224,
561795233, 561795242 during the academic year 2021-2022, under my
guidance and direct supervision at the Department of Pharmacology, Excel
College of Pharmacy, Komarapalayam.
Prof.Dr.R.MANIVANNAN,M.Pharm.,Ph.D
Principal,
Excel College of Pharmacy,
Komarapalayam-638183.
Place :
Date :
DECLARATION
We hereby declare that the dissertation “Antioxidant Activity and
Preliminary Phytochemical Studies of Various Extracts of Agaricus
bisporus”, submitted to The Tamil Nadu Dr.M.G.R. Medical University,
Chennai, for the partial fulfillment of the degree of Bachelor of Pharmacy,
is a bonafide research work carried out during the academic year 2021-2022,
under the guidance and direct supervision of Mrs.SRIVIDHYA.V,
M.Pharm., Department of Pharmacology, Excel College of Pharmacy,
Komarapalayam.
We further declare that this is original and this dissertation has not been
submitted previously for the award of any other degree, diploma or any other
similar title. The information furnished in this dissertation is genuine to the
best of our knowledge.
Place :
Date :
Miss.DIVYADARSHINI.S Mr.GOKUL.R
Mr.MANI.D Mr.JERALD J JACOB

ACKNOWLEDGEMENT
I first place out beloved thanks on the foot of my Parents and the almighty
for making my research a grand success. The contributions of many different
people, in their different ways, have made this research work possible. I feel
immensely delighted in expressing my sincere thanks from the core of my
heart and a deep sense of indelible gratitude to the following personalities.
First and foremost I am highly grateful to Prof.Dr.A.K.Natesan,
Honorable Chairman, Excel Group of Institutions and
Dr.N.Madhankarthick, Vice Chairman, Excel Group of Institutions, for
providing me all the infrastructure and resources to take up and complete
this work.
I am extremely beholden to, my guru Dr.R.Manivannan, Professor and
Principal, for his constant insight, personal advice, valuable guidance,
countless serenity, moral support, constant encouragement, pain taking effort
in all stages of study and constructive suggestions throughout the tenure of
the present investigation.
I express my thank to my guide Mrs.SRIVIDHYA.V, Assistant professor,
Department of Pharmacology, for providing all facilities and support
during the course of my research work.

I take immense pleasure to thank all department HODs, faculties and
librarians for their guidance and continued support.
My deepest heartfelt thanks to all my U.Jegatheesh, K.Dhilip kumar,
G.Bharanidharan and P.Gnanasekar for their help, co-operation, motivation,
ideas and encouragement.
I also express my thank to my class mates, for their support and
encouragement during the project work.
I extend my sincere thanks to ALL OTHER BATCHMATES and JUNIORS
for their help during the research work.
Individuals, who have helped/supported/guided/encouraged me directly or
indirectly during the course of my research work.
Last but not least, My special lovable thanks to my MOTHER, FATHER,
BROTHER, AND SISTER for their continuous overwhelming support.

CONTENT
S.NO TOPICS PAGE NO.
1 INTRODUCTION 1
2 REVIEW OF LITERATRE 11
3 AIM AND OBJECTIVES 16
4 PLAN OF WORK 17
5 PLANT PROFILE 18
6 MATERIALS AND METHODS 22
7 RESULTS 33
8 DISCUSSION 43
9 CONCLUSION 46
10 REFERENCES 47
TABLE OF CONTENT
S.NO CONTENT PAGE NO.

1 INTRODUCTION
1.1 Anti-Oxidants 1
1.2 Working of Antioxidants 3
1.3 Mechanism of Action 4
1.4 Sources of Antioxidants 5
1.5 Functions of Antioxidants 6
1.6 Uses of Antioxidants 6
1.7 AGARICUS BISPORUS 7
1.8 Benefits of Agaricus bisporus 8
1.9 Medicinal uses of Agaricus bisporus 9
2 REVIEW OF LITERATURE
3 AIM AND OBJECTIVES
4 PLAN OF WORK
5 PLANT PROFILE
5.1 Plant Profile 18
5.2 Habit 19
5.3 Geographical Distribution 20
5.4 Chemical Constitution 21
6 MATERIALS AND METHODS
6.1 Materials used for the study 22
6.1.1 List of Glass wares 22
6.1.2 List of Instruments 22
6.1.3 List of Chemicals 22
6.2 Methodology 23
6.2.1 Plant Collection 23
6.2.2 Processing of Plant Material 23
6.2.3 Extraction of Plant Materials 24
6.3 Preliminary Phytochemical Studies 25
6.3.1 Test for Phenols 25
6.3.2 Test for Tannins 26
6.3.3 Test for Glycosides 26
6.3.4 Test for Alkaloids 27
6.3.5 Test for Flavonoids 28
6.3.6 Test for Triterpenoids and Steroids 28
6.4 In-vitro Antioxidant Study 29
6.4.1 DPPH Assay 29
6.4.2 FRAP Assay 31
7 RESULTS
7.1 Preliminary Phytochemical Analysis 33
7.2 In-vitro Antioxidant Studies 34
7.2.1 DPPH radical scavenging activity of EEAB 34
7.2.2 Ferric Reducing Antioxidant Assay of EEAB 35
7.2.3 DPPH radical scavenging activity of MEAB 37
7.2.4 Ferric Reducing Antioxidant Assay of MEAB 38
7.2.5 DPPH radical scavenging activity of HAEAB 40
7.2.5 Ferric Reducing Antioxidant Assay of HAEAB 41
LIST OF ABBREVIATIONS
S.NO. Abbreviations Expansion
1 ROS Reactive Oxygen Species
2 SOD Super-Oxide Dismutase
3 DNA Deoxyribonucleic Acid
4 RNA Ribonucleic Acid
5 LDL Low Density Lipoprotein
6 LPO Lipid Peroxide
7 CAT Catalase
8 A. Bisporus Agaricus bisporus
9 DPPH 2, 2 – diphenyl-1-picrylhydrazyl Assay
10 FRAP Ferric Reducing Antioxidant Power Assay
11 nM nano Meter
12 w/w weight by weight
13 Ml Milliliter
14 EEAB Ethanol Extract of Agaricusbisporus
15 MEAB Methanol Extract of Agaricusbisporus
16 HAEAB Hydro Alcohol Extract of Agaricusbisporus

LIST OF TABLES
S.NO. TABLES PAGE.

NO.
1. Preliminary Phytochemical Analysis 33
2. DPPH radical scavenging activity of EEAB 34
3. Ferric Reducing Antioxidant Assay of EEAB 35
4. DPPH radical scavenging activity of MEAB 37
5. Ferric Reducing Antioxidant Assay of MEAB 38
6. DPPH radical scavenging activity of HAEAB 40
7. Ferric Reducing Antioxidant Assay of HAEAB 41

LIST OF FIGURES AND GRAPHS
S.NO. FIGURES PAGE.

NO.
1. Antioxidant Activity 2
2. Working of Antioxidants 4
3. Agaricusbisporus 7
4. Parts of Agaricusbisporus 19
5. Soxhlet Extraction Process 24
6. DPPH Assay Reaction (Purple 519nm turns into Colourless) 30
7. FRAP Assay Reaction 32
S.NO. GRAPHS PAGE.

NO.
1. DPPH Scavenging activities of EEAG and Standard Quercetin 35
2. Standard graph of vitamin C to determine the ferric reducing power of 36

EEAB
3. DPPH Scavenging activities of MEAG and Standard Quercetin 38

MEAB
5. DPPH Scavenging activities of HAEAG and Standard Quercetin 41

HAEAB
INTRODUCTION
INTRODUCTION
ANTI-OXIDANTS:
Anti-oxidants are compounds that act as a protective mechanism in the body,
neutralizing the effects of ROS (Reactive Oxygen Species) comprises of both free
radical and non-free radical oxygen containing molecules such as hydrogen peroxide
(H2O2), superoxide, singlet oxygen, the hydroxyl radical and inhibiting the oxidation
reaction which is a chemical reaction that can produce free radicals (Free radicals are
unstable, oxygen-containing molecules or atom that cause illness and aging that can
causes cell damage). Free radicals can produce huge chain chemical responses
in body, as well as the chain reactions damage body cells, because they react so easily
with other molecules. [1]
Enzymatic and Non-enzymatic sources are both possible. SOD (Super-Oxide
Dismutase – the first detoxifying enzyme and most powerful anti-oxidant in the cell),
catalase, glutaredoxin, and glutathione reductase are examples of enzymatic sources
that produce anti-oxidants. Vitamin C, vitamin E, selenium, zinc, beta carotene,
taurine, hypo-taurine, and glutathione are non-enzymatic sources of
antioxidants.However, as people become older, their antioxidant levels drop, causing
a disruption in the balance between antioxidants and pro-oxidant molecules, which
leads to oxidative stress. The scavenging antioxidant capacity, either due to a decrease
in antioxidant availability or an increase in ROS production[2, 3]
Department Of Pharmacology,Excel College Of Pharmacy. |1

INTRODUCTION
Figure 1. Antioxidant Activity
Excessive ROS produce oxidative stress by attacking bases in nucleic acids,
amino acid side chains in proteins, and double bonds in unsaturated fatty acids,
damaging DNA, RNA, proteins, and lipids, increasing the risk of cardiovascular
disease, cancer, and other disorders.Supplementing with oral oxidants can assist to
mitigate the effects of oxidative stress. Beta-carotene, vitamin A, and vitamin E
supplements had little influence on death rates or cancer risk. Furthermore, taking
selenium or vitamin E supplements does not lessen the risk of cardiovascular disease.
[4]
Oxidative stress occurs when your body's free radicals and antioxidants are out
of equilibrium. Because free radicals react so quickly with other molecules, they can
create enormous chain chemical reactions in your body. Oxidation is the term for
these kinds of processes. They have the ability to be useful or harmful. Antioxidants
are compounds that have the ability to give an electron to a free radical while
remaining stable. As a result, the free radical becomes less reactive and stabilizes. [5]

INTRODUCTION
ROS is produced by endogenous and external sources. Inflammation processes
and immune cell activation, extreme exercise, ischemia, mental activity stress,
malignant and infectious disorders, and ageing are all examples of endogenous
creation of these species. Exogenous sources of ROS include water and air pollution,
alcohol consumption, smoking, heavy metals, radiation, cooking, and certain solvents
such as benzene. After penetrating the body, these chemicals breakdown into ROS.
The destructive effects of reactive oxygen species (ROS) on cellular macromolecules
such as proteins, lipids, and nucleic acid are generating protein and nucleic acid
changes.[8]
Workings of Antioxidants:
Antioxidants are substances that can neutralize free radicals by receiving or
giving electrons, hence removing the radical's unpaired state. Antioxidant molecules
may directly react with reactive radicals and kill them, or they may transform into
new free radicals that are less active, have a longer lifespan, and are less hazardous
than the radicals they have neutralized. Other antioxidants or other mechanisms may
be able to neutralize them, putting an end to their radical condition. [6]

INTRODUCTION
Figure 2. Working of Antioxidant
Mechanism of Action:
Antioxidant principles are based on one of two mechanisms: chain breaking or
elimination of reactive oxygen species (ROS).The first is a chain-breaking mechanism
in which the principal antioxidant gives an electron to the system's free radical. The
second step includes quenching the chain-initiating catalyst, which removes
ROS/reactive nitrogen species initiators (secondary antioxidants). Antioxidants can
influence biological systems through a variety of processes, including electron
donation, metal ion chelation, co-antioxidants, and gene expression modulation. [7]

INTRODUCTION
Sources of Antioxidants:
 Vitamin A (Retinol), which is derived from beta-carotene and is also
generated by the body, preserves dark green, yellow, and orange vegetables
and fruits.
 Vitamin C (Ascorbic Acid) is a water-soluble molecule that has multiple
roles in living systems. Citrus fruits (oranges, sweet limes, and so on), green
peppers, broccoli, green leafy vegetables, black currants, strawberries,
blueberries, seabuckthorn, raw cabbage, and tomatoes are all good sources.
 Vitamin E is fat soluble and preserves lipids, including tocotrienol and
tocopherol. Wheat germ, seabuckthorn, nuts, seeds, whole grains, green leafy
vegetables, kiwifruit, vegetable oil, and fish liver oil are all good sources of
omega-3 fatty acids. The most common type of vitamin E taken is alpha-
tocopherol. Some tocotrienol isomers have been shown to have strong anti-
oxidant capabilities in recent investigations.
 Carotenoids, such as beta-carotene and lycopene, are found in apricots,
asparagus, beets, and broccoli.Collard greens, oranges, peaches, pink
grapefruit, pumpkin, winter squash, spinach, sweet potato, tangerines,
tomatoes, and watermelon are some of the fruits and vegetables that can be
used in a salad.
 Zinc in Beef, poultry, oysters, shrimp, sesame seeds, pumpkin seeds,
chickpeas, lentils, cashews, fortified cereals, sesame seeds, pumpkin seeds,
chickpeas, lentils, cashews
 Quercetin (apples, red wine, onions), catechins (tea, chocolate, berries),
resveratrol (red and white wine, grapes, peanuts, berries), coumaric acid
(spices, berries), anthocyanins.[9]

INTRODUCTION
Functions of Antioxidants:
 It lowers the amount of LDL-cholesterol, thereby reducing plaque deposition
in the blood vessel
 It is effective in cancer prevention.
 It encourages the proliferation of normal cells.
 Helps combat age-related molecular degeneration
 Antioxidants neutralize chemicals that can damage genetic material through
oxidation
 Supports the body's immune system[10]
Uses of Antioxidants:
 Treatment for cancer
 Treatment for high level Cholesterol
 Deep vein thrombosis
 Liver illness that develops
 Digestion issues
 Preventing Heart disease
 Preventing stomach ulcers
 Boosting the immune system.[11]

INTRODUCTION
AGARICUS BISPORUS
Agaricus bisporus originated in Brazil, but is now grown in China, Japan, and
Brazil.Tumor, high blood sugar, excessive cholesterol, artery hardening, continuing
liver disease, circulatory diseases, and digestive difficulties are all treated with the
agaricus mushroom. Heart illness, weakening bones, and stomach problems are
among the other uses. It's also used to help with physical and emotional stress, as well
as to improve the immune system. Extracts of the agaricus mushroom have been
approved as a food ingredient in Japan.
Agaricus is a fungus that grows on damp wood and decomposes organic
materials such as humus, horse dung, and other organic matter. It is fairly abundant on
the grassy lands throughout the summer when the weather is rainy. [12]
Figure 3.Agaricusbisporus
White Button Mushroom (A. bisporus) is another name for Agaricus bisporus
(A. bisporus). It is the most common wild and cultivated edible mushroom,

INTRODUCTION
accounting for more than 40% of all mushroom production worldwide. It is grown in
more than 70 nations and on every continent except Antarctica. A. bisporus has a
delectable flavor with higher nutritional content, as well as a pleasant aroma or
flavoring taste, and is utilized as food and in the food industry. It's thought to have a
lot of biological activity, isn't harmful, and has a lot of meaning.It is used in
fragrance, cosmetics, and pharmaceutical industries, in addition to food and drinks.
[13]
The flavour and texture of wild A. bisporus have many primary and secondary
metabolites have been identified as having therapeutic potential in the prevention and
treatment of diseases, including cancer, hyperlipidemia, microbial diseases,
cardiovascular difficulties, liver diseases, and immunological problems. A. bisporus is
a litter-degrading basidiomycete that is widely found in humid-rich habitats and has
been cultured on a large scale for the food industry as a model organism. It produces a
number of enzymes for detoxification and breakdown of humidified plant litter as a
result of its ecological niche. [14]
Benefits of Agaricusbisporus:
Cancer fighting properties:
Polyphenols, polysaccharides, ergothioneine, glutathione, selenium, and
vitamin C are antioxidant substances that responsible for mushrooms' cancer-fighting
abilities. These antioxidants works to prevent the negative effects of oxidative stress,
which causes cell injury, and can speed up the ageing process as well as raise the risk
of heart disease and some tumours.

INTRODUCTION
Promote Heart Health:
Heart disease is highly associated to oxidative stress, inflammation, and high
cholesterol and triglyceride levels, and white mushrooms' ergothioneine and beta
glucan content may help minimize this risk. Beta glucan is a type of dietary fibre that,
when digested, forms a gel-like material that decreases blood cholesterol levels.
Triglycerides and cholesterol are thus trapped, blocking their absorption.
Ergothioneine may also help lower triglyceride levels after food
Blood sugar control:
White mushroom polysaccharides may help reduce blood sugar levels and
improve insulin resistance.
Improved gut health:
Their polysaccharides also serve as prebiotics, or food for the good bacteria in
your gut, which boost gut health. [16, 17, 18]
Medicinal uses of Agaricusbisporus:
Cardiovascular disease, diabetes mellitus, fungal and bacterial infections, immune
system disorders, and tumours, are being treated using A. bisporus extracts or
bioactive substances as antioxidants, anti-cancer, and anti-inflammation.
Anticancer:
Polysaccharides from mushrooms showed significant anti-cancer effect
against a variety of cancer cell lines. Basidiomycota is known for producing
therapeutic potential, which are related to its glucan and other polysaccharide content.
Polysaccharides are members of the beta-glucan family of chemicals, and they appear
to have anti-tumorigenic properties by enhancing cellular immunity.

INTRODUCTION
Anti-hyperlipidemic:
Hyperlipidemia, represented by increased levels of triglycerides or cholesterol,
is a dominant risk factor that contributes to the progression and development of
subsequent cardiovascular disease and atherosclerosis, which is one of the most
serious diseases in humans.
Anti-diabetic:
A. bisporus contains high levels of dietary fibers and antioxidants including
vitamin C, D, and B12; folates and polyphenols that may provide beneficial effects on
cardiovascular and diabetic diseases.
Antimicrobial:
Antimicrobial activity was found in A. bisporus extracts obtained with methyl
alcohol against bacteria, yeasts, and dermatophytes. The therapeutic and nutritional
properties of mushrooms have been studied extensively. Mushroom extract is used in
a variety of cosmetic products. Skin whitening has been shown to be possible by
reducing the activity of the tyrosinase enzyme. The cosmetic industry has been
working hard to isolate several key compounds from mushrooms, confirm their
aesthetic properties, and then employ them in cosmetic goods like lotions and
creams.[19, 20, 21]
Department Of Pharmacology,Excel College Of Pharmacy. | 10

REVIEW OF LITERATURE
Barros L.,et al (2008) in this study chemical, biochemical, and electrochemical
methods were used to test the antioxidant activity in five Agaricus sp. mushrooms.
Chemical tests determined their reducing power and radical scavenging activity, while
biochemical assays determined their lipid peroxidation inhibition capability; cyclic
voltammetry and differential pulse voltammetry were used to characterize the
mushrooms extracts electrochemically. All of the species were shown to exhibit
antioxidant activity, and electrochemical tests demonstrated the mushroom extracts
had similar electrochemical responses, implying a similar electro active chemical
composition and higher oxidation potentials than the standards (ascorbic and Gallic
acids). The most effective species was Agaricussilvaticus, which had the lowest EC50
values in chemical and biochemical testing and the highest antioxidant activity in
electrochemical assays. [26]
JeongSC.,et al (2010) the study has examined that consuming Agaricusbisporus
promotes anti-cholesterol emic and anti-glycemic responses in rats which is feed
with hyper cholesterol emic diet and rats with type 2 diabetes caused by
streptozotocin injection. The decrease in plasma glucose and triglyceride
concentrations, liver enzyme activity, alanine aminotransferase and aspartate
aminotransferase, and liver weight increases in Streptozotocin-induced diabetic male
Sprague Dawley rats which is given with Agaricusbisporus powder for 3 weeks. Oral
administration of Agaricus b. Powders to hyper cholesterol emic rats for 4 weeks led
in a major reduction in plasma total cholesterol and low-density lipoprotein.
Agaricusbisporus is abundant in dietary fibers and antioxidants including vitamin C,
D, and B12, as well as folates and polyphenols, which may help prevent
cardiovascular and diabetes problems. [32]

M Kozarsk., et al (2011) the study was conducted on four mushroom species
Agaricusbisporus, AgaricusBrasiliensis, Ganodermalucidum, Phellinuslinteus by hot
water extraction and ethanol precipitation to obtain a partial purified polysaccharides
in which varying quantity of α and β –glucans was identified by FT-IR in that four
mushroom species. The amount of total glucans in the extracts was shown to be
related to the EC50 values of the chelating and suppressing power abilities. In vitro
tests of polysaccharide extract immunomodulatory revealed that extracts from A.
Bisporus, A. brasiliensis fruiting bodies, and G. lucidum spores stimulate the
production of IFN-γ in activated human. [23]
DharumaduraiDhanasekaran., et al (2013) have reported that on the extracellular
synthesis method for the preparation of silver nanoparticles in water using the extract
of Agaricusbisporus, a naturally occurring edible mushroom, as reducing and
protecting agents. The silver nanoparticles were characterized by ultraviolet-visible
spectroscopy, Fourier transform infrared spectroscopy and X-Ray diffraction analysis.
The synthesized silver nanoparticles showed antibacterial activity against the multi-
[28]
drug resistant Gram positive and Gram negative bacterial pathogens.
H Ghahremani-Majd., et al (2015) the study comprises the chemical composition of
white button mushroom which contain a variety of secondary metabolites, including
phenolic compounds and ergothioneine, the compound which act as an excellent
antioxidant, also this study includes the comparison of the antioxidant properties, total
phenolic, ergothioneine and mineral contents of the most consumed strains of white or
brown colorAgaricusbisporus[22]
C Meircea., et al (2015) in the study methanol was used to extract polyphenols and
flavonoids from two edible mushrooms, Agaricusbisporus and Pleurotusostreatus,

and one non-edible mushroom, Fomesfomentarius. Spectrophotometric techniques
were used to quantify the amount of these substances in each extract. In vitro studies
were used to assess the antioxidant characteristics of each extract, including the 2,2-
diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay, 2,2'-azino-bis-3-
ethylbenzthiazoline-6-sulphonic acid (ABTS) assay, iron chelating activity, in which
the highest concentration of polyphenols was found in the Fomesfomentarius,
whereas the highest concentration of flavonoids was found in the Pleurotusostreatus.

[24]
Li HJ.,et al (2015) in this study 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl
radicals, metal chelating activity, and reducing power of Agaricusbisporus and its
methanolic, 55 % ethanolic, aqueous extracts, and crude polysaccharide were used to
analyze the antioxidant potential of Agaricusbisporus. The anti-hypoxic effect of
crude polysaccharide was then investigated. In comparison to the blank control,
polysaccharide enhanced survival time, lowered blood urea nitrogen and lactic acid
content, and decreased blood urea nitrogen and lactic acid content. They isolated the
polysaccharide and evaluated its molecular weight as well as monosaccharide
components (glucose, fructose and mannose) with such great activity. From to this
analysis, wild Agaricusbisporus has a lot of potential to be used as a natural food to
treat oxidative and hypoxic stress. [27]
Mustafa NadhimOwaid.,et al (2017) the study includes the cultivation of
Agaricusbisporuswhich is used for recycling wastes, waste paper, oat straw, waste
tea leaves, some water plants and others. A. bisporus has many usages in human
dietary and pharmaceutical fields due to its composition of essential amino
acids, fatty acids, carbohydrates, low calories, crude fibers, trace elements

and vitamins. Recently synthesized nanoparticles from Agaricusbisporus were

[29]
used to treat cancer, viral, bacterial and fungal diseases.
BozenaMuszynska.,et al (2017) the study involved varieties of A. bisporuswhich
determines the varieties of nutritional value in A. bisporus. Which examines the
content of organic compounds in different parts of the plant body where organic
compound were determined using reversed phased High Performance Liquid
Chromatography. The study also demonstrates that the brown varieties of A. bisporus
are the richest in the examined substances [30]
Ramos M., et al (2019) the study reported that Agaricusbisporus is becoming a more
significant food business in Europe, however this has resulted in an increase in the
number of by-products from their industrial production. The antimicrobial,
antioxidant, anti-allergenic, and nutritional chemicals for advancements such as
nutraceuticals, food additives, and cleaning products has been reviewed in this study.
Carbohydrates, proteins, dietary fiber, phenolic compounds, polyunsaturated fatty
acids, vitamins, and minerals are abundant in edible mushrooms Agaricusbisporus. [33]
AtilaF.,et al (2021) in this study they have investigated that mushrooms has
major source of a various nutrients as well as medicinal bioactive substances.
Agaricusbisporus has a variety of nutrients such as carbs, proteins, lipids, fibers,
minerals, and vitamins. The presence of active ingredients such as polysaccharides,
lipopolysaccharides, essential amino acids, peptides, glycoproteins, nucleosides,
triterpenoids, lectins, fatty acids, and their derivatives has been associated to anti-
microbial, anti-cancer, anti-diabetic, anti-hyper cholesterol emic, anti-hypertensive,
hepatoprotective, and anti-oxidant effects in these mushrooms. [31]

PopescuML.,et al (2017) in this study Agaricusbisporus, a wild growing mushroom
and Agaricusblazei, cultivated mushrooms were used for identifying their chemical
characteristics and antioxidant activity. Spectrophotometric, spectrometric, and X-ray
diffraction techniques were used to determine phytochemical screening. DPPH (2,2-
diphenyl-1-picrylhydrazyl), ABTS (2,2'-azinobis-(3-ethylbenzothiazoline-6-
sulfonicacid) free radicals, ferric reducing power, and ferrous ions chelating
characteristics were used to evaluate antioxidant activity. The total phenolic content
of wild-growing Agaricusbisporus and Agaricusblazei is identical. Wild-growing
Agaricusbisporus had the greatest calcium and magnesium. The antioxidant capacity
of cultivated Agaricusbisporus was low. [25]

AIM AND OBJECTIVES
AIM AND OBJECTIVES
AIM:
 The aim of this study is to evaluate the Preliminary Phytochemical screening
and Antioxidant activity of Edible part of Agaricus bisporus by using of
different solvents Ethanol, Methanol and Hydro-Alcohol.
OBJECTIVES:
 To extract Agaricus bisporus and perform the preliminary phytochemicals
and identify the phytochemical for antioxidant activity like DPPH and FRAY
assay methods
 To evaluate the In-vitro antioxidant activity of Agaricus bisporus by different
solvents of Ethanol, Methanol and Hydro Alcohol

PLAN OF WORK
PLAN OF WORK
 Review of Literatures
 Collection of Plant Materials (Edible Part of Agaricusbisporus)
 Extraction of Powdered Sample of Agaricusbisporus
 Preliminary Phytochemical Analysis
 Test for Phenols
 Test for Tannins
 Test for Glycosides
 Test for Alkaloids
 Test for Flavonoids
 Test for Terpenoids and Steroids
 In-Vito antioxidant Studies
 DPPH Assay
 FRAP Assay

PLANT PROFILE
PLANT PROFILE
Plant Name - Agaricusbisporus
Common Name - White Button Mushroom, Crimini mushroom
Family - Agaricaceae
VERNACULAR NAMES:
Tamil - Vellaipottankalan
English - White Button Mushroom
Hindi - Kukurmutta
TAXONOMICAL CLASSIFICATION:
Kingdom-Fungi
Phylum-Basidiomycota
Subphylum-Hymenomycotina
Class-Agaricomycetes
Subclass-Agaricomyycetidae
Order-Agaricales
Family-Agaricaceae
Genus-Agaricus
Subject-Agaricus bisporus (J.E. Lange) Pilat

PLANT PROFILE
Habit:
Figure 3. Parts of Agaricusbisporus
Agaricusbisporusis a fungus's fleshy, spore-bearing fruiting body that grows
above ground, on soil, or on its feeding supply.
Cap 3-16 cm, convex to widely convex or almost flat with age; dry;
smooth or with pressed-down fibers or tiny scales; white in some
variations, brown in others; smooth or with pressed-down fibers or
small scales; white in some types, brown in others.
Spore Print White
Spores Elliptical and smooth, measuring 5.5-8.5 x 4-6.5 mm. Basidia is a
two-spored fungus.
Bruising White
Gills Close together; pinkish to pinkish brown at first, then dark brown to
blackish.
Stripe 2-8 cm long; 1-3 cm thick; strong; more or less equal; smooth or
with little scales below the ring; white, frequently bruised reddish;
with a ring that fades with age.
Veil Missing

PLANT PROFILE
Mycelium Pale, longitudinally radical, cottony, and forms a thick, persistent
mycelial mat with age. A chemical that is poisonous to nematodes is
typically secreted by aged mycelium in yellowish to orangish
droplets
Table 1. Habits of Agaricus bisporus
GEOGRAPHICAL DISTRIBUTION:
The light brown hue of modern commercial Agaricus mushroom variants
came from the original light brown Color of the common Agaricus mushroom. The
white mushroom was found in 1925 at the Keystone Mushroom Farm in Coatesville,
Pennsylvania, growing in a bed of brown mushrooms. The white mushroom was
taken back to the farm's owner, Louis Ferdinand Lambert, who is a trained
mycologist. It was considered as a more appealing food item, similar to how white
bread was received, and it was cultivated and disseminated.
Cultures were cultivated from the mutant individuals, similar to the
commercial development history of the navel orange and Red Delicious apple, and
most of the cream-colored shop mushrooms available today are products of this 1925
accidental natural mutation.
An uncommon to rare mushroom in wild, in Britain and Ireland Agaricus
bisporusis found nearly always in grassland. This species occurs also in mainland
Europe and elsewhere in the world including parts of North America. [46, 47]

PLANT PROFILE
Chemical Constituents:
Agaricus bisporus Ethanolicextracts were used to examine samples. There
were 174 metabolic products found, with 13 significant metabolites ranging from 1.2
to 83 percent (w/w), 13 metabolites at 1% (w/w), and 148 metabolites less than 1%
(w/w).
AgaritineMycotoxin : Agaricus bisporus has a higher percentage of spore and Gills
accessible.
Selenium : The fresh complete sample has a higher content.[48]

MATERIALS AND METHODS
1. Materials used for the study:
1.1. List of Glass wares:
 50ml Beaker
 100ml Beaker
 Round Bottom Flask
 Stirrer
 Burette
 Test Tubes
 Conical flask
1.2. List of instruments:
 Mortar and Pestle
 Heating Mantle
 Soxhlet Apparatus
 Weighing Balance
 Water bath
1.3. List of Chemicals:
 Ethanol
 Methanol
 Hydro Alcohol
 Distilled Water
 Ferric Chloride
 Lead acetate
 Gelatin

 Sulphuric Acid
 Chloroform
 Benzene
 Ammonia
 Sodium Picrate Solution
 Glacial acetate solution
 Potassium Mercuric Iodide
 Potassium Bismuth Iodide
 Potassium Iodide solution
 Aqueous solution of Picric Acid
 Zinc Dust
 Hydrochloric Acid
 Sodium Chloride
 Acetic Anhydride
2. Methodology:
2.1. Plant Collection:
Edible part of Agaricusbisporuswere collected from the surrounding areas
of Salem, Tamilnadu,India during the month of August, 2021
2.2. Processing of Plant Material:
The collected Edible material (Edible part of Agaricusbisporus) were
cleaned and shade dried.The dried sample of that edible part was powdered.
The coarse powder was labelled and stored inair tight container for further
use.

2.2.1. Extraction of Plant Materials:
Soxhlet Extraction:
Powdered sample (100g) in Ethanol, Methanol and Hydro Alcohol were stored
in tight container. Extracted by Soxhlet Extraction method using Soxhlet
Apparatus
Figure 5.Soxhlet Extraction Process

Procedure:
The finely powdered Agaricus bisporus was placed in a paper bag and placed in
a soxhlet apparatus. The extraction solvent was placed in a flask, and the vapours
were condensed. The powder sample was in a filter paper bag and the extract poured
into it.The liquid content of the chamber was collected into a flask when the level of
liquid in the chamber went to the top of the syphon tube.This procedure was repeated
until the syphon tube was completely empty. The collected extract was then
evaporated to entirely eliminate the solvent, leaving crude extract in a primary or
semi-solid state.
2.3. PRELIMINARY PHYTOCHEMICAL SCREENING:
TESTFOR PHENOLS:
Ferric Chloride Test:
Add distilled water to the sample extract, then a few drops of 10% ferric chloride. 5
minutes at a boil. The presence of phenol is indicated by the colour development of
red, purple, blue, or green.
Lead Acetate test:
After the sample extract has been diluted in distilled water, add a few drops of lead
acetate solution. Boil for 10 minutes in a water bath. The creation of a yellow colour
indicates the presence of phenol. [36]

TESTFOR TANNINS:
Lead Acetate test:
1 ml of 10% lead acetate solution was added to 10 mg of sample extract. The
presence of tannins and phenolic chemicals is indicated by the formation of a yellow
or voluminous white precipitate.
Gelatin Test:
Add a few drops of 1% gelatin solution containing 10% sodium chloride to the 5g of
sample extract. The presence of tannins is shown by the formation of white
precipitate. [37]
TEST FOR GLYCOSIDES:
Borntrager’s Test:
Add a few drops of dilute sulphuric acid to the 3 ml sample extract. Bring it to a boil
and then filter it. Add an equal volume of benzene or chloroform to the cooled
filtrate and shake well. The organic layers are well-separated, and then ammonia is
added. The presence of glycosides is indicated by the presence of pink colour in the
ammonical layer.
Cardiac glycoside Test:
Add a few drops of ferric chloride solution and strong sulphuric acid to the sample
extract. The presence of glycosides is indicated by the formation of two layers, the
lower layer being reddish brown in colour and the upper layer being bluish green in
colour.

Baljet Test:
When a few drops of sodium picrate solution are added to the sample extract, the
colour of the solution changes from yellow to orange, indicating that glycosides are
present.
Keller Killian test:
Add 1 drop of 0.1 percent ferric chloride solution to 1 ml of sample and 2 ml of
glacial acetic acid. 1 ml concentrated sulfuric acid, added in drops to the above
mixture to create two layers. There are two layers formed: the upper layer is light
bright green in colour. The lower layer is clear and transparent (Sulfuric acid layer).
A reddish-brown ring around the junction.[37, 38]
TEST FOR ALKALODS:
Mayer’s test:
To the drug extract add few drops of Mayer’s reagent (Potassium Mercuric Iodide).
Formation of creamy colour precipitate indicates the presence of alkaloids.
Dragendraff’s Test:
Add a few drops of Dragendraff's reagent to the drug extract (Potassium Bismuth
Iodide). The presence of alkaloids is indicated by the formation of a reddish brown
or orange coloured precipitate.
Wagner’s Test:
Add a few drops of Wagner's reagent to the drug extract (Potassium Iodide
Solution). The presence of alkaloids is indicated by the formation of a reddish brown
colour precipitate.

Hager’s Test:
Add a few drops of Hager's reagent to the drug (Aqueous solution of picric acid).
The presence of alkaloids is indicated by the formation of a yellow-colored
precipitate. [39, 40]
TEST FOR FLAVANOIDS:
Zinc Hydrochloride reduction Test:
Zinc dust and strong hydrochloric acid are added to the sample extract. The presence
of flavonoids is indicated by the appearance of yellow, yellow-orange, or orange.
Alkaline reagent Test:
2 ml of a 2 percent sodium chloride solution should be added to the sample extract.
The presence of flavonoids is indicated by the formation of a bright yellow colour
that fades to colourless when 2 drops of dilute sulfuric acid are added. [41]
TEST FOR TERPENOIDS & STEROIDS:
Libermann-Burchard Test:
After adding a few drops of acetic anhydride to the sample extract, it was heated and
cooled. The sidewalls of test tubes were then sprayed with strong sulphuric acid.The
presence of steroids is shown by the formation of a brown ring at the junction of two
layers, and the upper layer turns green.The presence of terpenoids is indicated by the
formation of a strong red colour.
Salkowski Test:
In a test tube, dissolve the sample extract in 2 ml chloroform, then add an equal
amount of strong sulphuric acid and leave to stand for a while.The presence of

steroids is shown by the presence of red colour in the lower layer.The presence of
terpenoids is indicated by the presence of yellow colour in the lower layer. [42, 43]
2.4 In-Vitro Anti-Oxidant Activities
DPPH Assay Method:
Antioxidant activity can be measured in a variety of ways, both in vitro and in
vivo. Blois in 1958 devised this method with the purpose of evaluating antioxidant
activity utilizing a stable free radical in a similar manner. The DPPH-based technique
is likely the most widely used in vitro assay due to its simplicity, rapidity, and low
cost.By transferring hydrogen from other molecules, the stable free radical DPPH (2,
2-diphenyl-1-picryl-hydrazyl-hydrate) can be reduced. In vivo investigations, on the
other hand, allow for research in a physiological setting but need the use of animal
models, some of which are expensive and time-consuming. [43, 44, 45]
Based on the process by which antioxidants limit lipid oxidation, resulting in
DPPH radical scavenging and thus evaluating free radical scavenging capability, the
DPPH assay is used to predict antioxidant activity. Because the analysis just takes a
few minutes, the method is widely used. The DPPH free radical is exceptionally
stable, interacts with hydrogen-donating compounds, and has a UV visible absorption
maximum of 517 nm. The method is based on antioxidants scavenging DPPH, which
after a reduction process decolorizes the DPPH methanol solution. In this test, the
antioxidants' ability to decrease the DPPH radical is evaluated. [46, 47, 48]

Principle:
The DPPH free radical method (2, 2-diphenyl-1-picryl-hydrazyl-hydrate) is an
antioxidant assay that utilizes electron transfer to generate a violet solution in ethanol.
In the presence of an antioxidant molecule, this free radical, which is stable at room
temperature, is neutralized, resulting in a colourless ethanol solution. The DPPH assay
allows for a simple and quick spectrophotometric evaluation of antioxidants. [42]
Procedure:
0.5 ml of sample mixed with 3 millilitre of ethanol at last 0.3 ml of DPPH ethanol-
based solvent was added. After some time, the sample size is decreased and the colour
changes from deep violet to pale yellow. Absorbance at 517nm was identified after 1
hour 40 minutes
Reaction:
Figure 6. DPPH Assay Reaction (Purple 519nm turns into Colourless)

FRAP Assay Method:
Principle:
Axelrod et al.in 1976 created FRAP as a tool for studying protein mobility in
living cells. FRAP removes fluorescence from a specific area of a cell or tissue by
photo-bleaching it with strong laser light. Because FRAP requires fluorescent
molecules to move about freely in order to work, this location is usually a cell
membrane or a place where diffusion occurs, such as the nucleus. As bleached
fluorophores move out and healthy fluorophores from other locations move in, the
fluorescence in the bleached area steadily recovers, hence the name fluorescence
recovery after photo-bleaching. [49, 50]
The Ferric Reducing Antioxidant Power (FRAP) Assay is a simple, quick, and
precise method for measuring antioxidant capacity in a wide range of biological
materials. The assay is high-throughput and customizable, and it can evaluate
antioxidant capacities as low as 0.2 mM Fe2+ equivalents. The ferric reducing
antioxidant power (FRAP) assay uses antioxidants as reductants in a redox-linked
colorimetric process in which Fe3+ is reduced to Fe2+ at low pH, resulting in the
formation of a coloured ferrous-probe complex from a colourless ferric-probe
complex.[51, 52]
Procedure:
Threemillilitres of sample and 100 millilitres of samplewere aggressively mixed. For
4 minutes, the absorbance was measured at 593nm at 30 second intervals. For
calibration, aqueous solution with known Fe2+ concentrations in the range of 100-

1000mol/liter was utilized. The FRAP values are calculated using the regression
equation.[53]
Reaction:
Figure 5. FRAP Assay Reaction

RESULTS
RESULT:
Extractive Yield of Ethanol : 12 gram
Extractive Yield of Methanol : 8 gram
Extractive Yield of Hydro-Alcohol : 10 gram
S.NO Name of the Test EEAB MEAB HAEAB
1 Phenols + + +
2 Tannins + + +
3 Glycosides + + +
4 Alkaloids + - +
5 Terpenoids - + +
6 Flavonoids - - +
7 Steroids - + -
Table 1. Preliminary Phytochemical Analysis
“+” represents PRESENT
“-“represents ABSENT

RESULTS
In vitro Anti-oxidant Studies:
DPPH radical scavenging activity of EEAB:
Treatment Concentration Percentage of IC50 Value
(μg/ml) Inhibition (μg/ml)
Standard 10 14.5 94.9μg/ml
(Quercetin) 25 21.3
50 36.2
75 51.7
100 71.2
Sample (EEAB) 10 10.2 81.4μg/ml
25 17.5
50 31.8
75 45.2
100 61.6
Table 2.DPPH Scavenging activities of EEAB and Standard Quercetin

RESULTS
80
70
60
Concentration
50
40
Standard - Quarcetin
30
Sample - EEAB
20
10
0
0 20 40 60 80 100 120
Percentage of Inhibition
Graph 1. DPPH Scavenging activities of EEAG and Standard Quercetin
Ferric Reducing Anti-oxidant Powder Assay of EEAB:
Treatment Concentration (μg/ml) Absorbance (593nm)
Standard 20 0.1484
(Vitamin C) 40 0.1984
60 0.2158
80 0.2424
100 0.2943
Sample (EEAB) 100 0.1523
Table 3.Concentration and Absorbance of Standard Vitamin C and EEAB

RESULTS
0.35
0.3
0.25
Absorbance
0.2
Standard - Vitamin C
0.15
Linear (Standard - Vitamin C)
0.1
0.05
0
0 20 40 60 80 100 120
Concentration µg/ml
Graph 2. Standard graph of vitamin C to determine the ferric reducing power of
EEAB
The total antioxidant activity of FRAP assay 21.5μg of AAE per gm equivalent to
Vitamin C

RESULTS
DPPH radical scavenging activity of MEAB:
(Quercetin) 25 22.3
50 41.9
75 52.7
100 75.8
25 19.8
50 34.5
75 49.0
100 65.2
Table 4.DPPH Scavenging activities of MEAB and Standard Quercetin

RESULTS
80
70
Concentration 60
50
40
Standard - Quercetin
30 Sample - MEAB
20
10
0
0 20 40 60 80 100 120
Graph 3. DPPH Scavenging activities of MEAG and Standard Quercetin
Ferric Reducing Anti-oxidant Powder Assay of MEAB:
Standard 20 0.1484
(Vitamin C) 40 0.1984
60 0.2158
80 0.2424
100 0.2943
Table 5.Concentration and Absorbance of Standard Vitamin C and MEAB

RESULTS
0.35
0.3
0.25
Absorbance
0.2
0.15 Linear (Standard - Vitamin C)
0.1
0.05
0
0 20 40 60 80 100 120
Graph 4. Standard graph of vitamin C to determine the ferric reducing power
for MEAB
The total antioxidant activity of FRAP assay 25μg of AAE per gm equivalent to
Vitamin C

RESULTS
DPPH radical scavenging activity of HAEAB:
(Quercetin) 25 21.9
50 36.5
75 51.2
100 69.3
25 18.9
50 31.0
75 44.8
100 60.5
Table 6.DPPH Scavenging activities of EEAB and Standard Quercetin

RESULTS
80
70
60
50
Concentration
40
Standard - Quarcetin
30 Sample - HAEAG
20
10
0
0 20 40 60 80 100 120
Graph 5. DPPH Scavenging activities of HAEAG and Standard Quercetin
Ferric Reducing Anti-oxidantPowder Assay of HAEAB:
Standard 20 0.1484
(Vitamin C) 40 0.1984
60 0.2158
80 0.2424
100 0.2943
Table 7. Concentration and Absorbance of Standard Vitamin C and HAEAB

RESULTS
0.35
0.3
0.25
Absorbance
0.2
0.15
Linear (Standard - Vitamin C)
0.1
0.05
0
0 20 40 60 80 100 120
Graph 6. Standard graph of vitamin C to determine the ferric reducing power
for HAEAB
The total antioxidant activity of FRAP assay 18μg of AAE per gm equivalent to
Vitamin C

DISCUSSION
DISCUSSION:
Anti-oxidants are man-made or natural substances that may prevent or delay
some types of cell damage. It also found in many foods including Fruits and
vegetables. They also available as dietary supplements. Anti-oxidants are compounds
that act as a protective mechanism in the body, neutralizing the effects of ROS
(Reactive Oxygen Species) comprise both free radical and non-free radical oxygen
containing molecules such as hydrogen peroxide (H2O2), superoxide, singlet oxygen
(1/2O2), the hydroxyl radical and inhibiting the oxidation reaction, a chemical
reaction that can produce free radicals. A diet high in anti-oxidant may reduce the risk
of many diseases including heart disease and certain cancers. Flavonoids and other
polyphenolics, vitamin C and D and Carotenoids are the most common dietary
antioxidant.
Agaricus bisporus is an edible basidiomycete mushroom. It has two colour state white
and brown. Agaricusbisporus distributed in the Tibetan Plateau has high-stress
resistance that is valuable for breeding improvements. Agaricus bisporus Lange have
a rounded cap, intact veils and are free of darkening/ browning. The cap is typically
white, but there are brown capped strains. Agaricus bisporus compost of two different
substrate to form its fruit bodies, which it grows vegetatively and the poor nutrient
casing soil in which the suitable physical, chemical and biological conditions
stimulate the initiation process and fruit body production
Agaricus bisporus was extracted by using solvent such as Ethanol, Methanol, Hydro
alcohol by soxhlet extraction to identify an antioxidant activity. 150 gm of Agaricus
bisporus powder are used for Ethanol extract, and its percentage yield is found to be
6.66%. 150 gm of Agaricus bisporus powder are used for Methanol extract, and its

DISCUSSION
percentage yield is found to be 8%. 150 gm of Agaricus bisporus powder are used for
Hydro Alcohol extract, and its percentage yield is found to be 5.33%
Preliminary Photochemical test for the extraction of EEAB (Ethanolic Extarct of
Agaricus bisporus) has shown the presence of Phenol, Tannin, Glycosides and
Flavanoids. MEAB (Methanolic Extract of Agaricus bisporus) has shown the
presence of Phenols, Tannins, Glycosides, Terpenoids, and Steroids. HAEAB (Hydro
Alcholic Extract of Agaricus bisporus) has shown the presence of Phenols, Tannins,
Glycosides, Alkaloids, Terpenoids and Flavanoids, so it may show an antioxidant
activity.
DPPH (2, 2-diphenyl-1-picrylhydrazyl) assay is a stable molecule soluble in methanol
characterized by its deep violet colour with an absorption maximum at 515nm. DPPH
free radical scavenging is a acceptable mechanism for screening the antioxidant
activity of plant extract. The DPPH assay, violet colour DPPH solution is reduced to
yellow colour product. This free radical, stable at room temperature is reduced in the
presence of an antioxidant molecule giving rise to colourless ethanol solution. DPPH
assay is used to predict antioxidant activities by mechanism in which antioxidant act
to inhibit lipid oxidation.
The Extraction of DPPH assay is, In EEAB, the IC50 value of this extract is 81.4,
when compare to the standard Quercetin, it is identified as the extract has Antioxidant
activity. In MEAB, the IC50 value of this extract is 75.5, when compare to the
standard Quercetin, it is identified as the extract has Antioxidant activity. In HAEAB,
the IC50 value of this extract is 82.8, when compare to the standard Quercetin, it is
identified as the extract has Antioxidant activity. A relationship between IC50 of
DPPH scavenging activities among different extract of A. Bisporus. Antioxidant

DISCUSSION
activity exhibited to a certain extent was probably due to other antioxidant compound
present in these A. Bisporus besides the phenolic compound.
FRAP (Ferric Reducing Antioxidant Power) assay is a high-throughput, adaptable and
can detect antioxidant. FRAP is easy way which provides a quick, sensitive for
measuring antioxidant capacity of variable biological samples. FRAP is an
antioxidant capacity assay that uses Vitamin C as standard. This assay is often used to
measure the antioxidant capacity of food, beverages, and nutritional supplements
containing polyhenols. This assays measure the antioxidant potential in samples
through the reduction of ferric ion (Fe3+) to ferrous ion (Fe2+) by antioxidant present
in sample.
The Extraction of FRAP assays are, In EEAB, the total antioxidant activity of FRAP
assay 21.5µg of AAE per gram equivalent to Vitamin C. In MEAB, the total
antioxidant activity of FRAP assay 25µg of AAE per gram equivalent to Vitamin C.
In HAEAB, the total antioxidant activity of FRAP assay 18µg of AAE per gram
equivalent to Vitamin C, so it is identified as the extracts has antioxidant activity.

CONCLUSION
CONCLUSION
The knowledge of medicines based on natural products is increasing every day and
their usage has more benefits. In this study, Agaricus bisporus is used in treatment of
many diseases. This review article demonstrates the antioxidant properties of
Agaricus bisporus by using different extracts. From the result of present study it was
concluded that, when compare to EEAB, MEAB, the HAEAB shows the better
antioxidant potential in the extraction of Agaricus bisporus
Department Of Pharmacology, Excel College Of Pharmacy. | 46

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ANTIOXIDANT ACTIVITY AND PRELIMINARY

PHYTOCHEMICAL STUDIES OF VARIOUS EXTRACTS

THE TAMILNADU Dr. M.G.R MEDICAL UNIVERSITY

In partial fulfillment of the requirements for the award of degree of

Under the guidance of

Excel College of Pharmacy

This is to certify that the dissertation work entitled “Antioxidant Activity

and Preliminary Phytochemical Studies of Various Extracts of Agaricus

bisporus” submitted by the students bearing REG NO: 561795221,

561795224, 561795233, 561795242 to “The Tamil Nadu Dr. M.G.R.

Medical University”, Chennai, in partial fulfillment for the award of

Bachelor of Pharmacy was evaluated by us during the examination held

INTERNAL EXAMINER EXTERNAL EXAMINER

This is to certify that the dissertation “Antioxidant Activity and

Preliminary Phytochemical Studies of Various Extracts of Agaricus

bisporus” is a bonafide work done by REG NO: 561795221, 561795224,

561795233, 561795242 at the Department of Pharmacology, Excel College

of Pharmacy, Komarapalayam, in partial fulfillment for the award of

Bachelor of Pharmacy under the guidance and direct supervision of

Mrs.SRIVIDHYA.V, during the academic year 2021-2022.

This is to certify that the work embodied in this dissertation entitled

“Antioxidant Activity and Preliminary Phytochemical Studies of

Various Extracts of Agaricus bisporus”, submitted to “The Tamil Nadu

Dr.M.G.R. Medical University”, Chennai, in partial fulfillment to the

requirement for the award of degree of Bachelor of Pharmacy, is a

bonafide work carried out by REG NO: 561795221, 561795224,

561795233, 561795242 during the academic year 2021-2022, under my

guidance and direct supervision at the Department of Pharmacology, Excel

College of Pharmacy, Komarapalayam.

We hereby declare that the dissertation “Antioxidant Activity and

Preliminary Phytochemical Studies of Various Extracts of Agaricus

bisporus”, submitted to The Tamil Nadu Dr.M.G.R. Medical University,

Chennai, for the partial fulfillment of the degree of Bachelor of Pharmacy,

under the guidance and direct supervision of Mrs.SRIVIDHYA.V,

M.Pharm., Department of Pharmacology, Excel College of Pharmacy,

similar title. The information furnished in this dissertation is genuine to the

best of our knowledge.

Mr.MANI.D Mr.JERALD J JACOB

for making my research a grand success. The contributions of many different

immensely delighted in expressing my sincere thanks from the core of my

heart and a deep sense of indelible gratitude to the following personalities.

First and foremost I am highly grateful to Prof.Dr.A.K.Natesan,

Honorable Chairman, Excel Group of Institutions and

Dr.N.Madhankarthick, Vice Chairman, Excel Group of Institutions, for

providing me all the infrastructure and resources to take up and complete

I am extremely beholden to, my guru Dr.R.Manivannan, Professor and

Principal, for his constant insight, personal advice, valuable guidance,

countless serenity, moral support, constant encouragement, pain taking effort

in all stages of study and constructive suggestions throughout the tenure of

the present investigation.

I express my thank to my guide Mrs.SRIVIDHYA.V, Assistant professor,

Department of Pharmacology, for providing all facilities and support

during the course of my research work.

librarians for their guidance and continued support.

My deepest heartfelt thanks to all my U.Jegatheesh, K.Dhilip kumar,

G.Bharanidharan and P.Gnanasekar for their help, co-operation, motivation,

ideas and encouragement.

I also express my thank to my class mates, for their support and