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Biomedical Application and Properties of Aloe Vera: Name: Yugesh Ghimire
Biomedical Application and Properties of Aloe Vera: Name: Yugesh Ghimire
A Term Paper
Submitted for the partial fulfillment of the requirements of
Master of Science Degree Second Semester Examination in Chemistry
Submitted by:
Name: Yugesh Ghimire
Roll No: 13/074
T.U. Registration No: 5-2-0037-2077-2013
Submitted to:
DEPARTMENT OF CHEMISTRY
Tri – Chandra Multiple Campus
INSTITUTE OF SCIENCE AND TECHNOLOGY
TRIBHUVAN UNIVERSITY, KATHMANDU, NEPAL
June, 2019
RECOMMENDATION
This is to certify that the term paper entitled, "Biomedical application and
properties of aloe vera" has been carried out by Mr. Yugesh Ghimire, the student of
M.Sc. Chemistry Second Semester Roll No. 13/074 for the partial fulfillment of
requirement of Master of Science degree in chemistry under my supervision. To the
best of my knowledge, this work has not been submitted to any other degree in this
institute.
...................................
Supervisor
Asst. Prof. Dr. Arvind Kumar Pathak
Department of Chemistry
Tri-Chandra Multiple Campus
Ghantaghar, Kathmandu Nepal
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ACKNOWLEDGEMENT
I am grateful to all teachers and my friends. Specially thanks for my colleagues: Mr.
Santosh Sah for his support during the preparation of the term paper.
......................
Yugesh Ghimire
June , 2019
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ABBREVIATION
iii
ABSTRACT
Aloe vera is herb distributed throughout the world. The herb is used internally to
combat most digestive problems, including constipation, poor appetite, diabetes,
immune system enhancement. The plant leaves contains numerous vitamins, minerals,
en-zymes, amino acids, natural sugars and other bioactive compounds. The first
objective of this study was to investigate the anti-bacterial activities of Aloe Vera
using methanol extract of Aloe Vera gel (MEAG), ethanol extract of Aloe Vera gel
(EEAG) and acetone extract of Aloe Vera gel (AEAG). The second objective of this
study was to evaluate the anti-oxidant activities of AVES using reducing and
chelating activities of Aloe Vera. The inhibition efficiency of Aloe Vera extract
(AVES) increased from 21.5% up to 75.56% on increasing of its concentration from 5
mg/Kg to 40 mg/Kg. This result indicates that Aloe extract provided a natural
antioxidant by scavenging free radicals.
Keywords: Aloe Vera, antioxidant, Aloe Vera extracts (AVES), bioactive compound
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TABLE OF CONTENTS
Recommendation letter
Acknowledgement ii
Abstract ii
Abbrevation iv
Table of contents v
1. Introduction 1
2. Literature review 7
3. Experimental method 9
4. Results and discussion 11
5. Conclusion 16
References 18
v
List of figures
Page no
vi
CHAPTER ONE
1. INTRODUCTION
1.1 Introduction of Aloe vera
Aloe barbadensis miller generally referred to as aloe vera is a species of plant
belonging to the genus Aloe, is the Arabic name for the plant while vera means true or
genuine. As belonging to the family Liliace that originated in South Africa but have
been indigenous to dry subtropical and tropical climates including the southern USA,
aloe vera has been used for medicinal purposes in several cultures for millennia:
Greece, Egypt, Mexico, Japan, and China, and leaves of aloe vera are particularly
used an ointment for the skin. According to the Chinese pharmacopie, Aloe vera has a
cold and bitter nature. The herb is used internally to combat most digestive problems
including constipation, poor apetite, collitis, irritable bowel syndrome as well as
asthama diabeties, immune system enhancement, peltic ulcers [1].
Aloe Vera is a succulent plant, succulants are commonly xerophytes and
having storage tissue that hold maxium water ranging from 99-99.5% and 0.5-1.0%
sold materials that include water and fat soluble vitamins, minerals, enzymes, simple
and complex polysaccharides, phenolic compounds and organic acids. Studies on the
structural components of the A.vera plant leaf be portions, the rind was found to 20-
30% and the pulp 70-8-% of the whole leaf weight. On the dry weight basis, the
percentages of the rind and pulp represented as lipids (2.7%-4.2%) and that as
proteins (6.3% and 7.3%) only accounted for a minor fraction. [2].
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1.2 Characteristics of Aloe vera
1.2.1 Physical classification:
Membranes which are green and leathery. Beneath it a tough resistant covering is the
gel which itself as a compact, gelatinous mass with a transculant pearly aspect. The
leaves reach 30-60 cm long, erected green in color, covered with rounded specks and
crowded in a basal rosette. Aloe vera leaf is fatty and smooth. It contains two separate
juice materials, the yellow latex (exudates) refered as “Aloe juice” or “Aloe sap” and
the transparent mucilaginous gel (Aloe vera gel)[ 3].
1.2.2 Chemical classification
Plant of Aloe vera is unique itself that contains richest source of many chemical
compounds. Chemistry of plant demonstrated that there are more than 200 different
biologically active substances which are present different parts of plant bodies. Plants
of aloe vera leaves are made up of three layers which is given as
Green rind/cuticles
Sap
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cuticles and chloroplast which has protective function and in synthesis carbohydrates
and proteins [4].
Anthraquinones
There are twelve different types of anthraquinones are present in the sap of Aloe vera:
Aloinm Isobarbaloin, Antracene, Emodin, Ester of Cinnamonic acid, Chrysophanic
acid, Barbaloin, Anthranol, Aloetic acid, Aloe Emodin, Ethereal oil and Resistannol.
They work as natural painkillers, laxatives and analgesics, and they have powerful
antibacterial, antifungal and virucidal features[5].
Amino Acids
Amino acids are the basic unit of protein, which help in the manufacture and repair of
muscle tissue. The human body needs 8 essential ones among 22 amino acids. Aloe
vera provides 20 of 22 required amino acids and 7 of 8 essential ones which are not
synthesized in our body[5].
Enzymes
Enzymes are natural protein molecules with highly specialized catalytic functions in
biochemical reactions produced by all living organisms. Although like all other
proteins, enzymes are composed of amino acids, they differ in function in that they
have the unique ability to facilitate biochemical reactions without undergoing change
themselves. Some of the most important enzymes in Aleo Vera are: Peroxidase,
Aiiase, Catalase, Lipase, Cellulose, carboxypeptidase, Amylase and Alkaline
Phosphates.[5]
Vitamins
The plants contains many vitamins, including the important antioxidant vitamins A,
C, and F. vitamins B (Thiamins), niacin, vitamin B2 (riboflavin), vitamin B12,
choline and folic acid. Generally vitamin B12 responsible for the production of red
blood cells. Similarly, Folic acid helps to develop the new blood cells [5].
Minerals
Aloe Vera contains the following minerals: calcium, manganese, sodium, copper,
magnesium potassium, zinc, chromium, iron etc. maganesium lactates inhibits the
histidine decarboxylates and prevents the formation of histamine from the amino
acids, histamine. Similarly, Sodium ensures that the body fluids do not become too
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acidic or too alkaline. Copper also enables iron to work as oxygen carriers in the red
blood cells[6].
Lignin
This cellulose substance is found in the gel has no known medical properties except it
posses the property of penetrating the human skin[6].
Saponins
These form soapy lathers when mixed and agitated with water. They have been used
in detergents, foaming agents and contain antiseptic properties[6].
Sugars
Aloe Vera contains both monosaccharide, such as glucose and fructose, and
polysaccharides. Polysaccharides are the most important types of sugars. They aid in
proper digestion, maintain cholesterol levels, improve liver functions and promote the
strengthening of bones[6].
Sterols
Sterols are important anti-inflammatory agents. The ones found in Aloe vera are:
Cholesterol, Sitosterol, Campesterol, and Lupeol. These sterols contain antiseptic and
analgesic properties. They also have pain killing properties similar to aspirin[6].
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1.3 Application
1.3.1 Medicinal uses
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Role in wound healing
Aloe vera helps in wound healing in the following proposed ways [6]:
1. Keeps the wound moist
2. Increases migration of epithelial cells
3. Enhances collagen maturation
4. Reduces inflammation
Effects on immune system
Aloe vera is a well-known adaptogen that has the capability to boost body’s natural
immunity. It balances the body’s system by stimulating body’s defense and adaptive
mechanisms. It contains 90% rhodium and iridium (trace minerals) in the acemannan
which is one of the polysaccharides. It acts as a great stimulant and dramatically
increases the white blood cells or macrophages and T cells. Thus, immunomodulating
effects occur via activation of macrophage cells to generate nitric oxide, secrete
cytokines, and present cell surface markers. It helps the thymus gland to enlarge in
size by 40% and thereby increase the production of T cells [6]
Antimicrobial properties
Various constituents of Aloe vera like sulphur, salicylic acid, cinnamic acid, urea
nitrogen and phenol act as a team to prevent the growth of disease-causing
microorganisms and thus exhibit antimicrobial activity. They help to eliminate many
internal and external infections [6].
Antibacterial property
Aloe vera gel has a broad spectrum activity and is shown to be effective against both
Gram-positive and Gram-negative bacteria. The antibacterial effect is suggested to
enhance the wound healing process by eliminating the bacteria which are responsible
for producing inflammation. The inner-leaf gel from Aloe vera was shown to be
effective inhibiting the growth of Streptococcus and Shigella species in vitro. Studies
conducted with Aloe vera in toothpastes have suggested that Aloe vera in gel and
toothpaste forms were equally effective against many species like Candida albicans,
Streptococcus mutans, Lactobacillus acidophilus, Enterococcus faecalis, Prevotella
intermedia and Peptostreptococcus anaerobius. Enhanced antibacterial effect against
S. mitis by Aloe vera gel was also demonstrated in some studies [9].
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CHAPTER TWO
2. REVIEW OF LITERATURE
Aloe Vera is a plant have great significant contribution in both medicinal and
phytopharmacological uses .Various Scientists and Researchers are devoted towards
its application and its field of research. As we know that various natural
antimicrobials are derived from the barks, stems, leaves and flowers regarding to this,
Aloe Vera is a natural medicinal plant which highly considered to the field of
research. Due to its multidimensional properties it is multidiciplinary plant. It was
also claimed that many biological activities of this plant can be attributed to the
polysaccharides found in the leaf gel.
Yagi et al., 2003.[10] reported that Aloe Vera gel contains a glycoprotein with cell
proliferating-promoting activity, while it is noted that aloe vera improved wound
healing by increasing blood supply (angiogenesis), which increased oxygenation as a
result. Angiogenesis is the growth of new blood capillaries and a part of tissue
regeneration. A study showed that topical application of Aloe Vera gel reestablished
vascularity of burn tissue for a gunie pig, altough no specific constituens were
indentified but two years later,davand et al. reported that low molecular weight
component of freeze-dried aloe vera stimulated blood vessels formation in a chick
chorioallantioc membrane.
Baby joseph et al., 2010. [11] was presented the study and it demostrated that the
traditional and pharmacological and phytopharmocological poroperties of various
bioactive compounds present in the aloe vera. Similarly he and his co-workers also
concluded that the the plants contains many vitamins A, C, F, vitamin B , niacin ,
vitamin B2, choline and folic acid. The leaf pulp and liquid fraction of aloe vera act
against various microorganis. Meanwhile processed aloe vera gel lowred the blood
glucose level by decreasing insulin resistance and also lowered the triglyceride levels
in liver and plasma of mice.
Gao Yan et al,. (2018).[12] Illustrated that the summerization of the pharmacological
activities and clinical studies of aloe vera and various extracts, as well as its extensive
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application of food chemistry, and will also discuss the future prospects of biomedical
applications of this herb.
Antimicrobial assay: The antibacterial and antifungal properties of the extracts were
assessed by using modified antimicrobial assay utilising microtitre plate described by
Drummond and Waigh et al., Briefly, using aseptic techniques a single colony of
microbe was transferresd into a 100ml bottle of peptone water broth, capped, and
placed in incubator overnight at 35c. After 12-18h of incubation, using aseptic
prepation and the aid of a centrifuge, a clean samp;e of microbe was prepared. To
sterile microtitre plates, 100micro litre of peptone water broth is first added to all
wells, followed by 100 micro liter of microbial culture was further added. The
resulting mixtures were then left to incubate for 24h at ambient temprature. After
incubation, 40 micro litre INT(iodonitrotetrazolium) was added and left it for 20
minutes. The microplates were then assessed visually to determine the minimum
inhibitory concentration (MIC). Controls used for bacteria are Gentamicin,
Chloramphenicol, and Streptomycin while controls used for fungi are Ampicilin and
Amphoterich.
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CHAPTER THREE
3.2 Methodology (seperation of bio active ingredents from aloe vera leaves)
High peformance liquid chromatography or high liquid chromatography (HPLC) is a
technique that is used to separate a mixture of compounds to indentify or purify the
individual components of a mixture. It allows you to use a very much smaller particle
size for the column packing materials which give a much greater surface area for
interraction between stationary phase and molecules following phase. This allows
much better seperation of the components of the mixture. Other major improvement
over column chromatography concerns the detection methods which can used these
methods are highly automated and extremly sensitive. Low and high pressure liquid
chromatography based seperation techniques are most widely used for fraction,
purification and size determination. Ion exchange chromatography is a separation
technique based on the charge molecules. It is used for the seperation of charge
molecules from the neutral one. Gas chromatographhy (GC) is another widely
employed technique. The mobile phase is gas, instead of liquid as in LPLC or HPLC.
It is suitable for the separation of volatile compounds. For example, acetyl or methyl
groups on a polysaccharide can be measured by this method following saponification.
9
Other commonly used analytical methods include specific rotation, infra red (IR) and
nuclear magnetic resonance (NMR) spectroscopy. Improved chromatographic
methods for determination of bio active compounds from Aloe vera Leaves.
10
Antioxidant activity (%)
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MTCC646 Escherichia coli. Antibacterial activity was determined by the agar- well
diffusion method.
The Aloe extract was potent against three strains of Mycobacterium (M. fortium, M.
smeg-matis and M. kansasii). and a strong anti-mycobacterial activity against M.
tuberculosis as well as antibacterial activity against P. aeruginosa, E. coli, S. typhi.
Streptoccocus pyogenes and Sterptoccocus faecalis are two microorganism that have
been inhibited by Aloe vera gel. Glucomannan and acemannan have been proved to
accelerate wound healing, activating macrophages, stimulating immune system as
well as antibacterial and antiviral effects.
Aloe vera gel was bactericidal against Pseudomonas aeruginosa and acemannan
prevented it from adhering to human lung epithelial cells in amonolayer culture, a
processed aloe vera gel prepation inhibited the growth of fungus Candida albicans, The
antibacterial activity test of the aloe vera plant extacts against E.coli and bacillus was
carried out by the agar well diffusion method. The 20ml of sterilized Muller Hinton Agar
was poured into sterile petri and solidifiede . On the 100 micro litre of E.coli and Bacillus
were swabbed on the respective plates. The sedded agar was punched out with sterile corl
borer at equally spaced wells on the medium to make holes. The extract methanol,
ethanol, acetone and their combination were filled in the wells and incubated for 72h
at37ºc. The diameter of inhibitory zones measured in mm.
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CHAPTER FOUR
4. RESULT AND DISCUSSION
4.2 Metal Chelating Activity of Aloe Vera: Transition metal ions such as copper
and iron are important in the production of ROS by donating and accepting single
electrons during redox reactions, cycling from reduced to oxidized forms and back as
shown below (Bashir et al., 2011).
13
bonds with a metal to reduce the redox potential thereby stabilizing the oxidized form
of the metal ion. Thus, the AVES have a well-known capacity for iron binding which
related with its iron binding capacity.
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reducing ability of AVES was observed in the formation of Perl’s blue when Fe3+
reduced to Fe2+ by donating electrons/H atom to the acceptor like free radicals to
form stable bonds as shown below.
Fe3+ + 2OH H2O2 +Fe2+ H2O + O2
The reducing power of Aloe Vera extracts. In this assay, the yellow color of the test
solution changes to Perl’s blue when reduction of the Fe3+ complex to the ferrous
complex. The amount of Fe2+ complex formation was monitored by measuring the
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CHAPTER FIVE
5. CONCLUSION
The plant Aloe Vera has both medicinal and phytopharmacological uses such as anti-
oxidant, antimicrobial, antitumor, antidiabetic, wound healing etc. It is aslo found that
this plant is also used in different civilization period as for different purpose such as
home remedies, skin moisturizing and anti-ageing, digestive tract health, resulting the
plant is boon to mankind. Aloe Vera also uses for the prevention and treatment of
several diseases. Since it is easily available with low price, no side effects, and eco-
friendly with environment.
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6.Suggestion for further work
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Reference
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Aloe vera in the treatment of various ailments, Bioscience Biotechnology
Research Committee., 2016, 9(2), 273-288.
2. Kumar S. and Yadav J.P., Ethnobotanical and pharmacological properties of
Aloe vera: A review, Journal of Medicinal Plant Research, 2014, 8(48), 1387-
1398.
3. Chatterjee P. , Chakraborty B. and Nandy S., Aloe vera plant: review with
significant pharmacological activities, Mintage journal of Pharmaceutical &
Medical Sciences, 2013, 2,21-24.
4. Chandrasekar R. and Sivagami B., Aloe a Gel in a Cell, Int J Pharmacogn
Chinese Med, 2018, 2(5), 147.
5. Chandrasekaran K., Salman M., Chavan K. S., Aloe vera - A Herbal Panacea
for Periodontal Disease, EC Dental Science, 2018,7(10), 1749- 1758.
6. Dubey K. P., Singh K. S., Dangi A., Bachle D. and Kataria K. R., Aloe vera: a
wonderfull plant for pharmacological, cosmetic use, and some other aspect,
ejpmr, 2017,4(2), 399-408
7. Sahu K. P., Giri D. D., Singh R. and Pandey P., Therapeutic and Medicinal
Uses of Aloe vera: A Review, Pharmacology & Pharmacy, 2013, 4, 599-610
8. DerbeT., Yilma B. and Badasa S., Investigation of anti-bacterial and anti-
oxidant activities of aloe vera plant at arba minch, southern Ethiopia, World
Journal of Pharmacy and Pharmaceutical Sciences, 2015, 4(1), 185-195
9. Johnsona B. D., Shringi B. N., Patidar D.K., Chalichema N. S. and Javvadi
A.K., Screening of antimicrobial activity of alcoholic and aqueous extract of
some indigenous plants, Indo Global. J. Pharma. Sci., 2011, 1, 186-11.
10. Yagi A., Kabash A., Mizuno K., Moustafa S., Khalifa M., Radical Scavenging
Glycoprotein Inhibiting Cyclooxygenease-2 and Thromboxane A2 Synthases
from Aloe vera Gel, Planta Medica, 2003, 69(3), 269-71
11. Joseph. S. B. and Raj J., Pharmacognostic and phytochemical properties of
aloe vera linn – an overview, International journal and pharmaceutical
science and research, 2010, 4(2), 106-110
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12. Gao Y., lengkuok K., Jin Y. and Wang R., Biomedical Applications of Aloe
vera, Critical Reviews in Food Science and Nutrition, 2018, 1, 1549-7852.
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