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Saurashtra University 
     Re – Accredited Grade ‘B’ by NAAC 
     (CGPA 2.93) 
 
 
 
 
Chauhan, Sanjaykumar P., 2010, “Phytochemical and pharmacological 
screening of fruit of Opuntia Elatior Mill


 thesis PhD, Saurashtra University 
 
 
http://etheses.saurashtrauniversity.edu/id/eprint/782 
 
 
Copyright and moral rights for this thesis are retained by the author 
 
A copy can be downloaded for personal non-commercial research or study, 
without prior permission or charge. 
 
This thesis cannot be reproduced or quoted extensively from without first 
obtaining permission in writing from the Author. 
 
The content must not be changed in any way or sold commercially in any 
format or medium without the formal permission of the Author 
 
When referring to this work, full bibliographic details including the author, title, 
awarding institution and date of the thesis must be given.
  
 
 
 
 
 
 
 
 
 
Saurashtra University Theses Service 
http://etheses.saurashtrauniversity.edu
 
repository@sauuni.ernet.in 
 
© The Author

 
PHYTOCHEMICAL AND PHARMACOLOGICAL 
SCREENING OF FRUIT OF OPUNTIA ELATIOR MILL. 
 
A THESIS 
 
SUBMITTED TO 
 
SAURASHTRA UNIVERSITY 
 
RAJKOT 
 
 
FOR 
 
The Award of 
 
Doctor of Philosophy (Ph. D.) 
 
Under the 
 
FACULTY OF MEDICINE (PHARMACY) 
 
 
By 
 
Mr. Sanjaykumar Prakashbhai Chauhan
 
(M. Pharm) 
 
Smt. R. B. Patel Mahila Pharmacy College 
Kailashnagar, Bhavnagar road, 
At: Atkot, Ta: Jasdan, Dist: Rajkot 
Gujarat 
 
 
 
RESEARCH GUIDE 
 
Dr. N. R. SHETH 
M. Pharm, Ph. D, L.L.B., D.B.M. 
Professor & Head, 
Department Of Pharmaceutical Sciences, 
Saurashtra University
Rajkot 
 
 
December, 2010 

 
PHYTOCHEMICAL AND PHARMACOLOGICAL 
SCREENING OF FRUIT OF OPUNTIA ELATIOR MILL. 
 
A THESIS 
 
SUBMITTED TO 
 
SAURASHTRA UNIVERSITY 
 
RAJKOT 
 
 
FOR 
 
The Award of 
 
Doctor of Philosophy (Ph. D.) 
 
Under the 
 
FACULTY OF MEDICINE (PHARMACY) 
 
 
By 
 
Mr. Sanjaykumar Prakashbhai Chauhan
 
(M. Pharm) 
 
Smt. R. B. Patel Mahila Pharmacy College 
Kailashnagar, Bhavnagar road, 
At: Atkot, Ta: Jasdan, Dist: Rajkot 
Gujarat 
 
 
 
RESEARCH GUIDE 
 
Dr. N. R. SHETH 
M. Pharm, Ph. D, L.L.B., D.B.M. 
Professor & Head, 
Department Of Pharmaceutical Sciences, 
Saurashtra University, 
Rajkot 
 
 
December, 2010 

Certificate 
 
 
This is to certify that the research work embodied in this thesis entitle,
 

Phytochemical and Pharmacological screening of fruit of Opuntia 
elatior Mill.

 
was carried out by Mr. Sanjaykumar Prakashbhai 
Chauhan at Smt. R. B. Patel Mahila Pharmacy College, Atkot under 
my guidance and supervision. This work is up to my satisfaction. The 
work embodied in this thesis is original and no part of the thesis has 
been submitted previously to this university or to any other university 
for the award of Ph. D. or any other degree or diploma.   
 
 
Guided by 
 
 
 
Dr. N. R. Sheth 
M. Pharm., Ph. D., L.L.B., D.B.M. 
Professor & Head, 
Department of Pharmaceutical Sciences, 
Saurashtra University,  
Rajkot. 
 

DECLARATION 
 
 
I, hereby declare that the thesis entitled Phytochemical and 
Pharmacological screening of fruit of Opuntia elatior Mill.”  is 
genuine record of research work carried out by me under the guidance 
and supervision of Dr. N. R. Sheth Professor & Head, Department of 
Pharmaceutical Sciences, Saurashtra University, Rajkot. No part of 
thesis has been submitted to any University or Institution for the 
award of any degree or diploma. 
 
 
 
 
 
 
 
Rajkot                               Mr. Sanjaykumar Prakashbhai Chauhan 
December, 2010                          
          M. Pharm. 
  Smt. R. B. Patel Mahila Pharmacy College, 
  Atkot.                                                        
 

 
COPYRIGHT 
 
 
DECLARATION BY THE CANDIDATE 
 
I hereby declare that the Saurashtra University, Rajkot shall have 
the right to preserve, use and disseminate this dissertation / thesis 
in print or electronic format for academic / research purpose. 
 
 
 
 
  Mr. Sanjay P. Chauhan 
  Ph. D. Reg. No. – 3627 
  Department of Pharmaceutical Sciences, 
  Saurashtra University,  
  Rajkot. 
Date:  
Place: Rajkot 
 
 
© Saurashtra University, Rajkot 
 

ACKNOWLEDGEMENTS 
 
 
A single flower can not make a garden or a single star can not make the 
beautiful shiny sky same way a research work can never be out come of a single 
individual’s talent or efforts. During my journey from objective to goal, I have 
experienced shower of blessings, guidance and inspiration from my teacher, parents, 
friends, all my well wishers and almighty God who have give me this seafaring right 
direction and made it to achieve the goal. Although it is not possible for me to name 
and thank them all individually, but it is great pleasure for me to pen down some of 
the special personalities who have made it possible for me to put this research work in 
present form. 
 
 
With my profound pleasure, I take this opportunity to express my deep sense 
of gratitude to my esteemed guide and mentor Dr. N. R. Sheth, M.Pharm, Ph. D., 
L.L.B., D.B.M., Proffessor & Head, Department of Pharmaceutical Sciences, 
Saurashtra University, Rajkot whose flowless guidance boundless enthusiasm, 
assiduous nature, enlightening discussion and valuable suggestions have enabled me 
to complete this work. I consider myself fortunate to have had the opportunity to work 
under his guidance. To work under the guidance of such an eminent person has been a 
great and inexplicable experience, which will go a long way down my memory have 
in my life. 
 
I am indebted to Dr. N. P. Jivani, Principal, Smt. R. B. Patel Mahila 
Pharmacy College, Atkot for continuous inspiration, guidance and providing facilities 
to complete my work. 
 
 
I would also like to express my deep sense of gratitude to Dr. B. N. Suhagia, 
Professor & Head, Dept. of Pharm. Chem., L.M. College of Pharmacy, Ahmedabad 
and Mr. I. S Rathod, M. Pharm, Assit. Professor, Dept of Pharm. Chemistry, L.M. 
College of Pharmacy, Ahmedabad who encouraged me and who has shown keen 
interest in my assignment and has inspired me and render selfless support throughout 
this investigation. 
 

With my profound gratitude I would like to thank my colleague of Smt. R.B. 
Patel Mahila Pharmacy College, Atkot. Mr. Ravi Manek, Mr. Jitendra Vaghasia, Mr. 
Shailesh Koradia, Mr. Shailesh Malaviya, Mr. Subhash Vaghani, Mr. Rishad Jivani, 
Mr. Tejas Serasiya, Mr. Tushar Gohil, Mr. Nimish Pathak, Mr. Ashok Parmar, Mr. 
Yagnik Bhalodia, Mr. Manish Goyani, Mr. S. Pandey, Mr. Piyush Patel, Mr. Viral 
Devmurari and rasik, rasilaben, nita, chhaya, ranjanben, yunusbhai and all other non-
teaching staff of college for encouraging and helping me a lot during my research 
work. 
 
 
I would like to express my deep sense of gratitude to Mr. Ashwin Dudhrejia, 
Mr. Mahesh Dabhi, Mr. Nilesh Patel and Mr. Mihir Raval, Dept. of Pharm. Sci., 
Saurashtra University, Rajkot for continuous guidance, direction and everlasting 
support enabled me to complete this research work. 
 
 
With my profound gratitude I would like to thanks Dr. S. Y.Gabhe, Principal, 
C.U. Shah College of Pharmacy, Mumbai for his continuous guidance, direction and 
ever lasting support enabled me to complete this project. His constructive cricism, 
perpetual encouragement, timely suggestion and meticulous attention were the real 
driving force throughout this project. 
 
 
I wish to thanks Bhatt Pathology Laboratory and Savani Kidney Hospital, 
Rajkot for helping to carried out my histopathological work.  
 
 
I also wish to thanks   Mr. Raju Dangar, Mr. Mehul Mer, Mr. Amit Modi, Mr. 
Vipul Mangrolia, Mr. C. Balakumar, Chetasbhai and all ABVP karyakarta for their 
sincere support and selfless help as and when required.   
 
 
I bow down with reverence and my heartfelt thanks to Mr. R.B. Patel (Bhagat 
Saheb), Prof. D.K Saiyad, and all the administrative staff of Smt. R.B. Patel Mahila 
Pharmacy College, Atkot for their excellent guidance, constructive suggestion, 
unstinted help, encouragement and flowless inspiration without which I would not 
have been able to accomplish this work. 
 
I acknowledge and dedicate my sincere gratitude towards my pappa Dr. 
Prakashbhai, mummy Vanitaben, my wife Hetal, my son Yajat and my loving brother 

Dr. Bhagirath for continuous prayers, inspiration and selfless help, who suffered a lot 
to keep me free from domestic responsibilities. My entire family members are the 
actual driving force that enabled me to reach this stage.  
 
 
Last but not least, I owe gratitude and thankfulness to all who were involved 
directly or indirectly, knowingly or unknowingly to reach in the venture of mine. 
 
 
 
                                                 
 
Date:        
 
Place:                                                                 Sanjaykumar Prakashbhai Chauhan 

 
 
 
Dedicated 
to  
My Family, 
Hetal, Yajat 
and 
My Guru 
 

Content 
 
 
Chapter Title  Page 
No. 
                Abbreviations 

                List of Table 
iii 
                List of Figure 
vi 
                Abstract 
xi 
1. Introduction 

2. 
Review of Literature 
13 
3. Materials 
and 
Methods 
89 
4. Results 
120 
5. Discussion 
197 
6. Summary 
216 
7. Reference 
220 
8. Annexure 
258 
 

Abbreviations 
 
AAS: Atomic Absorption Spectrophotometer 
ALP: Alkaline phosphatase 
BAW: n-Butanol:Acetic acid:Water 
CSF: Colony-stimulating factor 
DAD: Diode array detector 
EPO: Erythropoietin 
FLT-3 ligand: FMS-like tyrosine kinase 3 ligand 
G-CSF: Granulocyte-colony stimulating factor 
GM-CSF: Granulocyte Macrophage-colony stimulating factor 
Hb: Haemoglobin 
HgCl
2
: Mercuric chloride 
HPLC: High Performance Liquid Chromatographic 
HPTLC: High Performance Thin Layer Chromatographic 
i.p.: Intraperitoneal 
IDA: Iron Deficiency Anaemia 
IgE: Immunoglobulin E 
IL: Interleukin 
LC-MS: Liquid Chromatography – Mass Spectroscopy 
LTC
4
: Leukotriane C

LT-Rc: leukotriene receptor 
MCH: Mean Cell Haemoglobin 
MCHC: Mean Cell Haemoglobin Concentration 
MCP-2, 3, 4: Monocyte chemotactic protein-2, 3, 4 
M-CSF: Macrophage-colony stimulating factor 
MCV: Mean Cell Volume 
MDC: Macrophage-derived chemokine 
MIP-1α: Macrophage inflammatory protein-1α 
MPV: Mean Platelet Volume 
MS: Mass spectrometry  
NK: Natural killer 
 
i

OFJOpuntia elatior Mill. Fruit Juice 
p.o.: per oral 
PAD: Photodiode-array detector 
PAF: Platelet Activating Factor 
PCV: Packed Cell Volume / Haematocrite 
PDE: Phosphodiesterase 
PDW: Platelet Distribution Width 
PGD
2
: Prostaglandin D

PHZ: Phenylhydrazine 
RBC: Red Blood Cell 
RDW: Red blood cell Distribution Width 
rhEPO: recombinant hyman erythropeietin 
rhIL-2: recombinant human interleukin-2 
rhTPO: recombinant human thrombopoietic 
ROS: Reactive Oxygen Species 
SCF: Stem Cell Factor 
SDF-1: Stromal cell-derived factor-1 
TGF: Tissue Growth Factor 
TL: Total lipids 
TLC: Thin Layer Chromatographic 
TNF: Tumour Necrosis Factor 
TPO: Thrombopoietin 
TXA
2
 : Thromboxane A

WBC: White Blood Cell 
WHO: World Health Organization 
 
ii

List of Table 
Table 
No. 
Title Page 
No. 
1.1 
Anaemia prevalence affected in preschool – age children, pregnant 
women, and non – pregnant women in each region as per WHO. 

2.1 Haematopoietic 
cells, 
Growth 
factors and its analogues. 
16 
2.2 
Total phytochemical constituents of Opuntia spp. Cladodes. 
43 
2.3 
Minerals, Vitamins and Amino acids content in Opuntia spp. 
Cladodes. 
44 
2.4 
Pectin Content in Opuntia spp. Cladodes. 
46 
2.5 
Sugars, Hydrocolloids and Organic acids content in Opuntia spp. 
cladodes. 
46 
2.6 
Chief chemical constituents in fruits of Opuntia spp. 50 
2.7 
Mineral composition of prickly pear fruit peel. 
51 
2.8 
Lipids and sterols from peel of Opuntia spp. fruit. 
53 
2.9 
Evaluation of various samples of Opuntia ficus indica (L.) Mill. 
fruit juice obtained by ultrafiltration or osmotic distillation. 
54 
2.10 
The Mineral composition of cactus pear pulp. 
55 
2.11 
Mineral composition from O. dillenii and O. ficus indica. 55 
2.12 
Amino acid contents in fruit pulp of Opuntia spp. 56 
2.13 
Sugars (% of dry matter) and firbre (% of total fibre) composition 
in prickly pear fruit pulp. 
58 
2.14 
Flavonols, total flavonoids, ascorbic acid and total carotenoids 
content (µg/g fresh weight) in fruits of different Opuntia spp. 
61 
2.15 
Basic structure of anthocyanins and their absorption maxima. 
62 
2.16 
Qualitative analysis of betalains by HPLC. 
69 
2.17 
Qualitative data of betalains in prickly pear (Opuntia spp.) fruit by 
HPLC-ESI-MS. 
70 
 
iii

2.18 Spectrophotometric 
quantification 
of betalains in prickly pear. 
72 
2.19 
Spectrophotometric quantification of betalains in the fruits of 10 
Mexican prickly pear cultivars. 
73 
2.20 
Mineral composition of seeds of Opuntia ficus indica. 74 
2.21 
Amino acids composition of prickly pear seed flour and protein 
concentrate. 
75 
3.1 
Instrumental conditions for elemental analysis using AAS. 
98 
3.2 
List of organisms used for antimicrobial assay of peel extracts. 
118 
4.1 
Proximate analysis of fruit of Opuntia elatior Mill. 
122 
4.2 
Percentage yield and physical properties of the peel extracts. 
122 
4.3 
Presence of phytoconstituents in peel extracts and OFJ. 
123 
4.4 
Identification of carbohydrates from OFJ. 
124 
4.5 
Elemental analysis of OFJ by AAS. 
131 
4.6 
Measurement of body weight (g) of rats in HgCl
2
 –induced 
anaemia. 
133 
4.7 
Estimation of haemoglobin content (gm %) of rats in HgCl
2
  –
induced anaemia. 
134 
4.8 
Estimation of haemoglobin content (gm %) of rats in PHZ –
induced anaemia. 
150 
4.9 
Analgesic effect of OFJ on acetic acid –induced writhing in rat. 
177 
4.10 
Percentage inhibition of OFJ and diclofenac sodium on acetic acid 
–induced writhing in rat. 
178 
4.11 
Effect of OFJ and tramadol on pain induced by tail immersion test. 
180 
4.12 
Percentage analgesic action (Mean ± SEM) of OFJ on pain 
induced by tail immersion test. 
180 
4.13 
Effect of OFJ on preconvulsion time of guinea pig after histamine 
and acetylcholine aerosol exposure. 
181 
4.14 
Effect of OFJ on Histamine and Acetylcholine induced 
Bronchospasm in guinea pigs. 
 
181 
 
iv

4.15 
Effect of OFJ on acetylcholine –induced percentage maximum 
contraction of isolated rat ileum. 
182 
4.16 
Effect of OFJ on histamine –induced percentage maximum 
contraction of isolated guinea pig ileum. 
184 
4.17 
Effect of OFJ on egg albumin induced rat mast cell degranulation. 
186 
4.18 
Effect of OFJ on compound 48/80 induced rat mast cell 
degranulation. 
187 
4.19 
Effect of OFJ on carrageenan induced rat paw edema over 5 hour. 
189 
4.20 
Percentage inflammation produced by carrageenan in rat paw.  
189 
4.21 
Effect of OFJ on carrageenan – induced rat paw edema at 3 hour. 
190 
4.22 
Effect of OFJ on neutrophil adhesion. 
192 
4.23 
In vitro antimicrobial activity order of fruit peel extracts of 
Opuntia elatior Mill. 
193 
 
 
v

List of Figures 
 
Figure 
No. 
Title Page 
No. 
1.1a 
Anaemia as a public health problem by country: Preschool – 
age children. 

1.1b 
Anaemia as a public health problem by country: Pregnant 
women. 

1.1c 
Anaemia as a public health problem by country: Non – 
pregnant women. 

1.2 
The prevalence of asthma in the world. 

2.1 
Development of cells of the Haematopoietic systme. 
17 
2.2 
Haematopoietic growth factor development. 
18 
2.3 
Normal breakdown of Haemoglobin. 
21 
2.4 
Asthmatic pathway from intrinsic and extrinsic stimulus. 
27 
2.5 
Pathogenesis and drug action in asthma. 
30 
2.6 
Early-phase and Late-phase responses in asthma. 
31 
2.7 
Role of eosinophils in the late asthmatic reaction. 
34 
2.8 
The plant shape of Opuntia spp. 
37 
2.9 Habitus 
of 
Opuntia spp. 
38 
2.10 Phylloclades 
Shape. 
38 
2.11 
Shape of Fruit of Opuntia spp. 
39 
2.12 
Recepticular Scar Position in Fruit of Opuntia spp. 
40 
2.13 
Structures of Opuntisterol & Opuntisteroside. 
48 
2.14 
Phenolic compounds from cladodes of Opuntia spp. 49 
2.15 
Chemical structures of organic acids in cactus pear fruit and 
phylloclade. 
59 
2.16 
Structures of betacyanins and betaxanthins found in prickly 
pear. 
64 
2.17 
Biosynthetic scheme of betacyanin and betaxanthin formation. 
65 
4.1 
Photographs of plant Opuntia elatior Mill. (A) and Herbarium 
(B). 
121 
 
vi

4.2 
Macroscopic characters of phylloclade (A), flowers (B), ripe 
and unripe fruits (C) and transverse and longitudinal sections 
of fruit (D) of Opuntia elatior Mill. 
121 
4.3 
Comparison of R
f
 values of standard sugars and OFJ. 
125 
4.4 
Identification of sugar from fruit juice using Thin Layer 
Chromatography (TLC). 
126 
4.5 
Photograph of thin layer chromatography (TLC) of OFJ in 
BAW (A) and 1% aqueous HCl (B). 
127 
4.6 
Visible light absorption spectra of water (red line), methanol 
(black line) and methanol HCl (blue line) extracts of fruits of 
Opuntia elatior Mill. 
127 
4.7 
HPLC chromatogram of methanolic extract of fruits of 
Opuntia elatior Mill. 
128 
4.8 
Positive ion spray mass spectra corresponding to betanin, 
parent ion scan of m/z  551 (A); daugther ion scan of m/z  389 
(B). 
129 
4.9 
Calibration curve for total sugar content by anthrone reagent 
method. 
130 
4.10 
Calibration curve for gallic acid by Folin Ciocalteu method. 
130 
4.11 
Calibration curve for titratable acidity equivalent to citric acid. 
131 
4.12 
Effect of OFJ on body weight (g) in HgCl
2
 –induced anaemia. 
133 
4.13 
Effect of OFJ on Hb content (gm %) in HgCl
2
 –induced 
anaemia. 
135 
4.14 
The percentage recovery in Hb content of rats in HgCl2 –
induced anaemia at day 60. 
135 
4.15 A 
Effect of OFJ on total RBC (A)  and  PCV  (B) in HgCl
2
  –
induced anaemia. 
137 
4.15 B 
Effect of OFJ on MCV (C) and MCH (D) in HgCl
2
 –induced 
anaemia. 
138 
4.15 C 
Effect of OFJ on MCHC (E) and RDW (F) in HgCl
2
 –induced 
anaemia. 
139 
4.16 
Effect of OFJ on total WBC (A), Neutrophils (B) and 
Lymphocytes (C) in HgCl
2
 –induced anaemia. 
140 
 
vii

4.17 
Effect of OFJ on mean platelet count (A), PDW (B) and MPV 


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