Devendra Nath final thesis ppt.ppt for st

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1. 1

A Thesis on
PHYTOCHEMICAL INVESTIGATIONS AND SCREENING FOR
ANTIOXIDANT AND ANTIMICROBIAL ATTRIBUTES OF PSIDIUM
GUAJAVA ROOT EXTRACTS
Submitted to
GLOCAL UNIVERSIT
In Partial Fulfillment of the Requirements for the Degree of Master of Pharmacy
Batch- 2021-2023
Presented by
DEVENDRA NATH
(Reg. No. GU21R0562)


DEPARMENT OF PHARMACOLOGY, GLOCAL SCHOOL OF PHARMACY
GLOCAL UNIVERSITY, MIRZAPUR POLE, SAHARANPUR, UTTAR PRADESH-
247121
SUPERVISOR
Prof. (Dr.) Satish Kumar
Sharma
Professor & Principal
Glocal School of Pharmacy,
Glocal University
CO-SUPERVISOR
Mr. Ashok Kumar
Associate Professor
Glocal School of Pharmacy,

2. 2
Introduction
Aim and Objective
Plan of work
Results and Discussion
Conclusion
References
CONTENTCONTENT

3.  G
lobally, food spoilage caused by microorganisms still widely affects all types of food and causes food
waste and loss, even in developed countries.
 I
t has been estimated that the yearly losses of global food reach up to 40% due to various factors including
spoilage by microorganisms
 B
acteria, yeast, and molds are the common types of microorganisms responsible for the spoilage of a
considerable number of food and food products
 O
nce these microorganisms reach food products, they grow by utilizing the nutrients and produce
metabolites that cause food spoilage
 M
icroorganisms are available naturally in the surrounding environment; therefore they can easily reach food
during harvesting, slaughtering, processing, and packaging
 T
he understanding of the mechanism of antimicrobial action of medicinal plants extracts is the first step in
the optimal utilization of these extracts as natural antimicrobial agents to extend the shelf-life and maintain
the food quality.
 A
ntimicrobial agents are essentially important in reducing the global burden of infectious diseases.
INTRODUCTION

4.  H
owever, emergence and dissemination of multidrug resistant (MDR) strain in
pathogenic bacteria have become a significant public health threat as there are
fewer, or even sometimes no, effective antimicrobial agents available for the
infection caused by pathogenic bacteria.
 T
hus, in the light of the evidence of the rapid global spread of resistant clinical
isolates, the need to find new antimicrobial agents is of paramount
importance.
 A
vast number of medicinal plants have been recognized as valuable resources
of natural antimicrobial compounds as an alternative that can potentially be
effective in the treatment of these problematic bacterial infections.
 A
ccording to the World Health Organization (WHO), medicinal plants would be
the best source to obtain a variety of drugs
 
4

5. Mechanisms of Antimicrobial Resistance
5

6. 6

7. PSIDIUM GUAJAVA: PLANT PROFILE
• G
uava,
 
Psidium guajava 
(Linn.), a member of 
Myrtaceae 
family, is a
common tropical plant with a long history of traditional usage.
• I
t is used not only as food but also as folk medicine, and various parts of
this plant have a number of medicinal properties.
7A. Ripen fruits of P. guajava; B. Roots of P. guajava
A B

8. • P
. guajava 
is mainly known for its antispasmodic and antimicrobial
properties in the treatment of diarrhoea and
 dysentery.
• H
as also been used extensively as a
 hypoglycaemic agent.
• M
any pharmacological studies have demonstrated the ability of this
plant to exhibit antioxidant, anti-allergy, antimicrobial,
antigenotoxic, antiplasmodial, cytotoxic, antispasmodic,
cardioactive, anticough, antidiabetic, antiinflamatory and
 ,
supporting its traditional uses.
8

9. Major Chemical constituents
9

10. C
ollection, authentication and extraction of Psidium guajava roots.
S
tandardization of raw herb as well as their prepared extracts.
I
n-vitro Scavenging activity by DPPH method
T
o investigate the standardized extract of Psidium guajava roots for
their Antimicrobial activity.
10
Aim and Objective

11. Collection and Authentication of selected plant
Physicochemical parameters
Preliminary Phytochemical test
H.P.T.L.C. fingerprinting analysis
Determination of TPC and TFC
Pharmacological activity
In-vitro Scavenging activity by DPPH method
Antimicrobial assay of extracts
Preperation of microbial culture
Determination of Minimum Inhibitory Concentrations
Statistical Analysis
Presentation of Thesis
11
Plan of Work

12. •C
ollection and
Authentication
•T
he Psidium guajava
roots were collected in
December from BHU
Varanasi.
•R
eference number: Myrta.
2023/1. 12
Results

13. Physicochemical analysis of Psidium guajava roots
S. No.Particulars Percentage (% w/w)
1.Loss on drying 3.9
2.Total ash 5.9
3.Water soluble ash 4.5
4.Acid insoluble ash 0.9
5.Sulphated ash 2.2
6.Pet. ether soluble extractive value 1.5
7.Ethyl acetate soluble extractive value2.0
8.Methanol soluble extractive value 5.1
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14. 14
Phytochemical test Pet. Ether extractEthyl acetate Extract Methanol extract
Alkaloid test   
Hager’s test - + ++
Mayer’s test - - +
Carbohydrate test   
Fehling’s test + ++ -
Barfoed’s test -  +
Molish’s test + ++ -
Benedic’s test + - +
Fixed oils and fats   
Filter paper test ++ - +
Peptides, Amino acids   
Biuret’s test - + +
Ninhydrin test - + +
Millon’s test + + +
Flavonoid test   
FeCl3 test - ++ ++
Alkaline reagent test + - ++
Lead acetate test - + ++
Zinc chloride acid reduction test - - ++
Shinoda’s reagent + + ++
Test for sterols   
Salkowaski test ++ - -
Test for saponin glycosides   
Foam test - + ++
For Starch   
Iodine test - ++ 
tannic acid - - ++
For tannins and phenolic compounds   
Ferric chloride solution + - ++
Phytochemical screening of Psidium guajava roots extracts

15. HPTLC Quantitative Fingerprinting Analysis
15
HPTLC Chromatogram of
standard Quercetin (0.1 mg/ml)
and methanolic extract of Psidium
guajava roots (10 mg/ml) at 365
nm. (Quantify: 0.0016µg/ml)
HPTLC Chromatogram of the
methanolic extract of Psidium
guajava roots at 365 nm.
Mobile Phase: Toluene: Ethyl
acetate: Formic acid (5:4:1).
The R
f
value is find out as 0.19,
0.29, 0.36, 0.43, 0.43, 0.54, 0.59,
and 0.63

16. Total Phenolic and Flavonoid Content
16

17. Total Phenolic and Flavonoid content
Extracts Total phenol content
(mg/g of gallic acid)
*
TFC
(mg/g Quercetin) *
Pet. ether extract of
Psidium guajava
1.341±0.063 0.627±0.46
Ethyl acetate extract of
Psidium guajava
4.23±0.042 5.208±0.26
Methanolic extract of
Psidium guajava
7.78±0.063 13.155±0.12
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18. DPPH Scavenging activity
18
Antioxidant capacity of Methanolic extract with reference to Gallic acid

19. Inhibition %age of extracts of Psidium guajava roots (MPG)
by DPPH assay
19

20. Antimicrobial assay of Methanolic extract of
Psidium guajava root disc diffusion method
20
Microorganism ZOI (cm)
50μg 100μg 200μg 400μg Ampicillin
10μg
Flucanazole
10μg
B. subtilis 0.8±0.30b 1.2±0.10a1.6±0.10b2.6±0.20b18±0.00a-
S. aureus - 0.5±0.00c1.0±0.00d1.5±0.05f19±0.05b-
S. epidermidis - 0.8±0.00c1.0±0.05e1.8±0.15c17±0.11a-
Proteus sp. - 1.3±0.11a1.6±0.10a2.0±0.10b18±0.11a-
Salmonella sp. - 0.8±0.05f1.3±0.00e1.7±0.00e16±0.00a-
E. coli 0.2±0.00c 0.8±0.11c1.3±0.11d2.1±0.55a19±0.00a-
C. albicans 0.8±0.20b 1.3±0.30b1.5±0.35b1.8±0.25a - 17±0.05b

21. 21

A

B

C

D

E

F
G
Whereas
B. subtilis; b. S.
aureus; c. S.
epidermidis; d.
Proteus mirabilis; e. S.
typhimurium; f. E.coli;
g. C. albicans

22. MIC Assay
U. ramosaB. subtilis
Gram +ve
S. aureus
Gram +ve
S.
epidermidis
Gram +ve
Proteus
mirabilis
Gram –ve
Salmonella
typhimurium
Gram -ve
E. coli
Gram –ve
MIC values in µg/ml
Methanol
extract
300 145 378 300 245 245
Ampicillin 15 10 10 15 10 10
22
Microtiter plate showing MIC values
for various microorganisms

23. Conclusion
 T
he current investigation examined the antimicrobial
 as well as antioxidant capacity of a MEPG roots.
Physicochemical evaluation, fluorescence analysis, Chemical tests, and anti-oxidant activity were carried out.
The HPTLC analysis for the qualitative as well as quantitative findings of constituents present in Psidium
guajava roots was performed.
 I
n the present study, the standardisation of the plant was done by various pharmacognostic technique which were
mainly used for the correct identity of the plant. The plant after the identification has been investigated for ash
values, powder analysis, extractive values etc. The results of the pharmacognostic studies features that underline
the correct identification and scientific validation of the Psidium guajava roots.
 T
he present investigation revealed the presence of steroids, triterpenoids, oils and fats, flavonoids, coumarins,
tannins as well as phenolic components.
 I
n the current research, root extract of Psidium guajava exhibited a promising antimicrobial and antioxidant
activity.The DPPH radical scavenging activity results of MEPG are comparable with known antioxidant
Vitamin C.
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24. 24
From the analysis it is observed that themethanolic extracts also showed maximum
antioxidant property indicating the free radical scavenging abilities of the plant.
Plant secondary metabolites are mostly responsible for their antimicrobial activity.
Major groups of phytochemicals which possess antimicrobial properties are
phenolics and polyphenols (flavonoids, quinones, tannins, coumarins), terpenoids,
alkaloids, lectins and polypeptides.
Based on the results (i.e., the antimicrobial activity), PE, EA and MeOH appeared
the best solvents for extraction of most of the plants in this investigation, whereas
hexane was least suitable. The standard drug was chosen as fluconazole and
ampicilline.
EA and MeOH were also seen as the optimum extraction solvents for some of the
active plants. Thus the Plant Psidium guajava roots may prove to be an important
antimicrobial and antioxidant agent.

25. Plagiarism Report
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26. References
1.Abdalla S, Apramian SS, Cantley LF, et al. Occupation and Risk for Injuries. In: Mock CN, Nugent R, Kobusingye O,
et al., editors. Injury Prevention and Environmental Health. 3rd edition. Washington (DC): The International Bank for
Reconstruction and Development / The World Bank; 2017 Oct 27. Chapter 6.
 Available from:
https://www.ncbi.nlm.nih.gov/books/NBK525209/
 doi: 10.1596/978-1-4648-0522-6_ch6.
2.Abdel-Wahhab MA, Ahmed HH, Hagazi MM. Prevention of aflatoxin B1-initiated hepatotoxicity in rat by marine algae
extracts. J Appl Toxicol. 2006 May-Jun;26(3):229-38. doi: 10.1002/jat.1127. PMID: 16389658.
3.Ahmad S, Zeb A, Khan S. Effects of aqueous extract of Medicago denticulata against paracetamol-induced
hepatotoxicity in rabbits. J Food Biochem. 2021 Dec;45(12):e13985. doi: 10.1111/jfbc.13985. Epub 2021 Oct 28.
PMID: 34713456.
4.Anbarasu, C., Rajkapoor, B., Bhat, K. S., Giridharan, J., Amuthan, A. A., & Satish, K. (2012). Protective effect of
Pisonia aculeata on thioacetamide induced hepatotoxicity in rats.
 
Asian Pacific journal of tropical biomedicine,
 
2(7),
511–515. https://doi.org/10.1016/S2221-1691(12)60087-2.
5.Asrani SK, Devarbhavi H, Eaton J, Kamath PS. Burden of liver diseases in the world. J Hepatol. 2019 Jan;70(1):151-
171. doi: 10.1016/j.jhep.2018.09.014. Epub 2018 Sep 26. PMID: 30266282.
6.Ayoub SS. Paracetamol (acetaminophen): A familiar drug with an unexplained mechanism of action. Temperature
(Austin). 2021 Mar 16;8(4):351-371. doi: 10.1080/23328940.2021.1886392. PMID: 34901318; PMCID: PMC8654482.
7.Azam, F. M., Biswas, A., Mannan, A., Afsana, N. A., Jahan, R., & Rahmatullah, M. (2014). Are famine food plants also
ethnomedicinal plants? An ethnomedicinal appraisal of famine food plants of two districts of bangladesh.
 
Evidence-
based complementary and alternative medicine : eCAM,
 
2014, 741712. https://doi.org/10.1155/2014/741712.
8.Chen YF, Jobanputra P, Barton P, Bryan S, Fry-Smith A, Harris G, Taylor RS. Cyclooxygenase-2 selective non-
steroidal anti-inflammatory drugs (etodolac, meloxicam, celecoxib, rofecoxib, etoricoxib, valdecoxib and lumiracoxib)
for osteoarthritis and rheumatoid arthritis: a systematic review and economic evaluation. Health Technol Assess. 2008
Apr;12(11):1-278, iii. doi: 10.3310/hta12110. PMID: 18405470.
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27. Thank You
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