Antioxidant and Cytotoxic activity of crude extract of Opuntia elatior Mill Submitted By Shawon ...

85 Pages·2015·2.58 MB·English

Antioxidant and Cytotoxic activity of crude extract of
Opuntia elatior Mill

A Dissertation submitted for the partial fulfillment of the course of Pharmaceutical
Research (PHRM-404) of the Department of Pharmacy, East West University, for
the degree of Bachelor of Pharmacy.

Submitted By
Shawon Majumder
ID: 2011-3-70-040

Supervised by
Abdullah-Al-Faysal

Lecturer

Department of Pharmacy,
East West University, Dhaka.
Page | i
Declaration by the Research candidate
I,Shawon Majumder, hereby declare that this dissertation, entitled “Antioxidant and Cytotoxic
activity of crude extract of Opuntia elatior mill” submitted by me to the Department of
Pharmacy, East West University, in the partial fulfillment of the requirement for the degree of
Bachelor of Pharmacy (B.PHRM) is a complete record of original research work carried out by
me during the period 2014-2015 under the supervision and guidance of Abdullah-Al-Faysal,
Lecturer, Department of Pharmacy, East West University, Dhaka. And it has not formed the
basis for the award of any other Degree/Diploma/Fellowship or other similar title to any
candidate of any university.

--------------------------------
Shawon Majumder
ID: 2011-3-70-040
Department of Pharmacy
East West University
Aftabnagar, Dhaka

Page | ii
Certification by the Supervisor
This is to certify that the dissertation, entitled “Antioxidant and Cytotoxic activity of crude
extract of Opuntia elatior mill” submitted to the Department of Pharmacy, East West University,
in partial fulfillment of the requirements for the award of the degree of Bachelor of Pharmacy
(B.PHRM) is a complete record of original research work carried out by me during the period
2014-2015 under the supervision and guidance of Abdullah-Al-Faysal, Lecturer, Department of
Pharmacy, East West University, Dhaka. And it has not formed the basis for the award of any
other Degree/Diploma/Fellowship or other similar title to any candidate of any university.

---------------------------------------
Abdullah-Al-Faysal
Lecturer
Department of Pharmacy,
East West University, Dhaka

Page | iii
Endorsement by the Chairperson
This is to certify that the dissertation, entitled “Antioxidant and Cytotoxic activity of crude
extract of Opuntia elatior mill” submitted to the Department of Pharmacy, East West University,
in partial fulfillment of the requirements for the award of the degree of Bachelor of Pharmacy
(B.PHRM) is a complete record of original research work carried out by me during the period
2014-2015 under the supervision and guidance of Abdullah-Al-Faysal, Lecturer, Department of
Pharmacy, East West University, Dhaka. And it has not formed the basis for the award of any
other Degree/Diploma/Fellowship or other similar title to any candidate of any university.

---------------------------------------
Dr. Shamsun Nahar Khan
Associate Professor & Chairperson
Department of Pharmacy
East West University
Aftabnagar, Dhaka

Page | iv
Acknowledgement
At first I would like to thank my research supervisor Abdullah-Al-Faysal, Lecturer, Department
of Pharmacy, East West University for his constant support and understanding throughout the
progress of the project. Without his sincere help and supervision and valuable advice it would
not have been possible for me to accomplish this study.
I acknowledge to Dr. Shamsun Nahar Khan, Chairperson and Associate Professor, Department
of Pharmacy for giving me the opportunities to use Laboratory and other facilities and for
encouraging me to do this research work.
I would also like to extend my thanks to all the research students in the lab, lab officers and other
staffs of the Department of Pharmacy for their help and assistance, friendly behavior and earnest
co-operation which enabled me to work in a very congenial and comfortable ambience.
I owe special thanks to my fellow research group members for their immense support and
contribution in my research work.
Last, I am grateful to my family for their constant financial support and encouragement to
complete the research work.

Page | v
ABSTRACT
The purpose of the study was to evaluate the antioxidant and cytotoxic activity of crude extract
of Opuntia elatior mill (Family: Cactaceae).
The powdered of Opuntia elatior mill were extracted with methanol and then partitioned with
pet-ether, DCM, ethyl acetate and crude fraction was taken for experiment.
The crude extract was used to evaluate cytotoxic and antioxidant activities. The cytotoxic
activity was measured by brine shrimp lethality bioassay.
.Screening for cytotoxic properties using brine shrimp lethality bioassay with tamoxifen (LC
50
value of 25μg/ml) as positive control showed that the fraction have considerable cytotoxic
potency exhibiting LC50 value 6.25μg/ml.
The crude extract contained 23.895 mg AAE/g of total phenolic content, 111.1 mg AAE/g of
total reducing power content and 58 mg AAE/g total flavonoid content.
The results of study clearly indicate the presence of cytotoxic and antioxidant properties of crude
extract. The obtained results provide a support for the use of this plant in traditional medicine
and its further investigation.
Key words: Opuntia elatior mill, Brine shrimp lethality bio-assay, phenolic content, flavonoid
content, reducing power assay.

Page | vi
Table of content
Content page
1.1 Introductin 1
1.2 Research on Traditional drug in Bangladesh 2
1.3 Phytochemistry 3
1.4 Selection, collection and identification of plant material 4
1.4.1 Selection 4
1.4.2 Collection 4
1.4.3 Harvesting 5
1.4.4 Drying 5
1.4.5 Grinding 5
1.4.6 Extraction of plant materials 6
1.4.7 Choice of solvents 7
1.5 Medicinal Plants 9
1.6 Humans and Plants 9
1.7 Goals of using Medicinal plants as Therapeutic agents 10
1.8 Importance of Medicinal plants 10
1.9 Classification of Medicinal plants 11
1.10 Traditional Medicine 14
1.11 Medicinal plant in Bangladesh 16
1.12 Use of Medicinal plant in Bangladesh 18
1.13 Economic value 19
1.14 Approaches of drug development 19
Page | vii
1.15 Antioxidant activity 21
1.16 Plant review 23
1.16.1 Plant description 23
1.16.2 Taxonomy 24
1.16.3 Synonyms 24
1.16.4 Traditional use of opuntia elator Mill 25
1.17 Aim of this experiment 25
2 literature review of the biological activities of opuntia elator mill 27
2.1 Analgesic and anti-inflammatory action 27
2.2 Effects on mast cell degranulation 27
2.3 As an Eco –friendly inhibitor 28
2.4 Reversible antifertility effect 29
2.5 Ethnomedicinal value 29
2.6 Antioxident potential 30
2.7 Up regulation of CAM 30
2.8 Phytochemical study and evalution 31
2.9 Bronchodialatory properties 31
2.10 Haematinic Evalution 31
2.11 Evalution of physiochemical properties 32
2.12 Histochemical studies in stomatal apparatus 33
2.13 Anti-ulcer activity 33
Page | viii
3.1 Plant collection, powdering, extraction and filtration 34
3.1.1 collectin 34
3.1.2 Process of powdering 34
3.1.3 Extraction 34
3.1.4 Filtration 34
3.1.5 Evaporation and extract preparation 35
3.2 Partition of mother solution 36
3.2.1 Partition with pet-ether 36
3.2.2 Partitioning with DCM 37
3.2.3 Partitioning with ethyl acetate 37
3.2.4 Collection of crude extracts 39
3.3 Antioxidant activity 39
3.3.1 Total Phenolic content 39
3.3.2 Principle 39
3.3.3 Apparatus and Reagents 40
3.3.4 Procedure 41
3.3.4.1 Standard curve preparation 41
3.3.4.2 Sample preparation 41
3.3.5 Determination of total phenol content 41
3.4 Total Flavonoid content 42
3.4.1 Principle 42
Page | ix
3.4.2 Apparatus and Reagents 42
3.4.3 Procedure 42
3.5 Reducing power assay 45
3.5.1 Principle 45
3.5.2 Apparatus and Reagents 45
3.5.3 Procedure 45
3.5.4 Standard solution preparation 46
3.5.5 Extract solution preparation 46
3.5.6 Determination of reducing power 47
3.5.7 Calculation of total reducing power 47
3.6 Brine Shrimp Lethality Bioassay 48
3.6.1 Principle 48
3.6.2 Apparatus and Reagents 48
3.6.3 Procedure 49
3.6.3.1 Preparation of sea water 49
3.6.3.2 Hatching of Brine Shrimp 49
3.6.3.3 Preparation of Test solution 50
3.6.3.4 Preparation of Test sample 50
3.6.3.5 Preparation of positive control Group 50
3.6.3.6 Preparation of Negative control Group 50
3.6.3.7 Counting of Nauplii 51
Page | x