PROTOCOL FOR
M.PHARM DISSERTATION
SUBMITTED TO THE
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
BANGALORE, KARNATAKA.
BY
MUHAMMED RIYAS
M.PHARM PART - I
DEPARTMENT OF PHARMACEUTICAL BIOTECHNOLOGY
2008-2009
UNDER THE GUIDANCE OF
Dr. T. TAMIZH MANI Ph.D.,
PROFESSOR ,
DEPARTMENT OF PHARMACEUTICAL BIOTECHNOLOGY,
BHARATHI COLLEGE OF PHARMACY,
BHARATHI NAGARA.
KARNATAKA-571422
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
BANGALORE, KARNATAKA.
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / Name of the Candidate and Address(In Block Letters) / MUHAMMED RIYAS,
MANGADATH HOUSE
MANATHALA-PUNNA
P.O CHAVAKKAD 680506
TRISSUR (DT) KERALA
2. / Name of the Institution / BHARATHI COLLEGE OF PHARMACY,
BHARATHI NAGARA.
3. / Course of Study and Subject / MASTER OF PHARMACY IN PHARMACEUTICAL BIOTECHNOLOGY.
4. / Date of Admission of Course / 30-06-2008
5. / Title of Topic / CHARACTERIZATION AND ANTIMICROBIAL PROPERRTIES OF HERBAL FORMULATIONS
6. / Brief Resume of the Intended Work
6.1 Need for the study
6.2 Review of the literature
6.3 Objectives of the study /
ENCLOSURE-I
7 /
Materials and Methods
7.1 Source of data
7.2 Method of collection of data
7.3 Does study require any investigations or interventions to conduct on patients or other human or animal?
7.4 Has ethical clearance been obtained from your institution in case of 7.3
/ENCLOSURE-II
8 / List of References / ENCLOSURE-III
ENCLOSURE-I
6. BRIEF RESUME OF THE INTENDED WORK:
6.1 Need For The Study:
It is estimated that today, plant materials are present in, or have provided the models for 50% Western drugs. The primary benefits of using plant derived medicines are that they are relatively safer than synthetic alternatives, offering profound therapeutic benefits and more affordable treatment.1
Among the several medicinal plants, Citrullus colocynthis, Tephrosia villosa, Bauhinia tomentosa which are available at certain specific areas like Western Ghats (Tamil Nadu) and Wayanad (Kerala) have got many phytochemical values as well as pharmacological activities. For example, Citrullus colocynthis is used medicinally for its drastic purgative, cathartic action2 and also it is used as a hepatoprotective3. Tephrosia villosa leaf juice is used to treat dropsy, diabetes4 and has got anti-microbial activity. Bauhinia tomentosa is found to have anti oxidant and anti-inflammatory activity as well as cancer cell growth inhibitory action.5
Plant based antimicrobials represent a vast untapped source for medicines. Continued and further exploration of plant antimicrobials needs to occur. Plants based antimicrobials have enormous therapeutic potential. They are effective in the treatment of infectious diseases while simultaneously mitigating many of the side effects that are often associated with synthetic anti-microbials. They are effective, yet gentle6. So in this present research programme anti-microbial study of the herbal formulation prepared from the above three plants will be undertaken.
While collecting the plants, people who are unaware of the difference between healthy and diseased plants will mix up the diseased plants along with the healthy plants during the production of herbal formulations. This may not lead to the production of effective formulations, as these formulations may have lesser medicinal values and higher toxicity. So in this present research programme both the healthy and diseased plants and seeds from different regions will be collected and from the collected plants different formulations will be prepared and then a phytochemical study will be undertaken which will help to create awareness among the people about the quality difference between healthy and diseased plants. From this formulation isolation and characterization, qualitative analysis by Gel Electrophoresis and study of anti-inflammatory activity is also planned with experimental programmes.
6.2 Review Of Literature:
1. Qureshi R et al., Reported that Citrullus colocynthis commonly called as wild gourd is a well known medicinal herb. It produces purgative action due to the presence of alkaloids. Hence it is well recognized as a medicinal herb and its main action is on the alimentary tract. This plant is commonly used for the digestive complaints in human beings2.
2. Mukherjee A et al., Reported phytopharmacological studies of Citrullus colocynthis for its hepatoprotective activity against carbon tetrachloride induced hepatotoxicity in albino rats. Alcoholic and aqueous extract of root of Citrullus colocynthis exhibited significant hepatoprotective activity in carbon tetrachloride challenged albino rats3.
3. Balakrishnan BR et al., Reported the anti hyperglycemic effect of Tephrosia villosa was more pronounced in the diabetic animals, in which administration for 12 days after alloxan induced diabetes. After alloxan induced diabetes it was observed that both standard and aqueous extract of Tephrosia villosa were significantly superior to control in reducing blood sugar on long treatment (12) days4.
4. Zainul Amiruddin Zakaria et al., Studied the Antinociceptive, Anti-Inflammatory and Anti-pyretic Properties of the Aqueous Extract of Bauhinia purpurea Leaves in Experimental Animals. This study showed that the aqueous extract of Bauhinia. purpurea leaves possesses potential pharmacological activities that require further investigation and, thus, confirms the folklore use of the plant in the treatment of ailments associated with pain and inflammation5.
5. Rani et al., Reported that anti microbial evaluation of some medicinal plants for their anti-enteric potential agent multi drug resistant salmonella typhi. Strong anti-bacterial activity was shown by methanol extract of Aegle marmelos, Salmalia malabarica, Tephrosia purpurea Punica granatum, Terminalia arjuna7.
6. Khouri et al., Studied the effect of Citrullus colocynthis (Cucurbitaceae) aqueous extract in 70% ethanol on blood biochemical parameters after oral administration of the extract to albino rats in a dose of 500mg/kg body weight for 7 days. Results demonstrated that administration of aqueous extract induces significant decrease in total serum cholesterol level8.
7. Mythreye et al., Reported the anti-microbial activity of leaves of Bauhinia tomentosa, methanol extract has shown significant activity against the tested micro organisms in comparison with the standard doxycycline and ketoconazole9.
8. Pettit G.R et al., Reported isolation and structures of bauhiniastatins 1-4 from Bauhinia purpurea. Bauhinastatin 1-4 were found to exhibit significant growth inhibition against a minipanel of human cancer cell lines and found to inhibit lymphocytic cancer cell line10.
9. Vijayan et al., characterized a defensin, TvD1 from Tephrosia villosa. The open reading frame of the cDNA was 228bp, which codes for a peptide with 75 amino acids. Expression analyses indicated that this defensin is expressed constitutively in Tephrosia villosa with leaf, stem, root, and seed showing almost similar levels of high expression. The recombinant peptide (rTvD1), expressed in the Escherichia coli expression system, exhibited potent in vitro antifungal activity against several filamentous soil-borne fungal pathogens11.
6.3 Objectives Of The Study:
Ø Preparation of herbal formulation from three different plants.
Ø Isolation and characterization of phytochemicals
Ø Qualitative analysis of different varieties of plants.
Ø Anti-microbial study of herbal formulation
Ø Comparative phytochemical study of healthy and diseased plant formulations.
Ø Anti-inflammatory study of the herbal formulation
7. MATERIALS AND METHODS:
The present work undertakes the collection of Citrullus colocynthis, Tephrosia villosa, and Bauhinia tomentosa and will be subjected for the following experiments
Ø Preparation of aqueous extract by Cold maceration.
Ø Gel Electrophoresis
Ø Anti-microbial studies using different strains of gram positive and gram negative disease causing bacteria’s and fungus by agar diffusion and microbial assay methods.
Ø Anti-inflammatory study by using male albino rats with the help of plethysmograph apparatus.12
All the above mentioned tests shall be carried out at department of pharmaceutical biotechnology. The lab is adequately equipped with necessary set up to carry out the works. The following instruments such as Hot air oven, Incubator, Autoclave, Cool centrifuge, HPLC, HPTLC, PCR, Gel Documentation system etc are available in the college lab.
. 7.1 Source Of Data:
Ø Bharahti College of pharmacy library, Bharathi Nagara.
Ø E-library from Bharahti College of pharmacy library, Bharathi Nagara.
Ø IISC library, Bangalore.
.
7.2 Method Of Collection Data:
Data collected from the following Journals, books and related links.
JOURNALS:
Ø Medicinal and Aromatic Plant Abstract.
Ø Journal of Agricultural and Food Chemistry.
Ø Indian Journal of Biotechnology.
Ø Journal of Biosciences, Biotechnology Research Asia.
Ø Journal of Plant Diseases.
Ø Journal of Natural Products.
Ø Indian Journal of Pharmaceutical Education and Research.
Ø Journal of Applied Microbiology and Biotechnology
BOOKS
Ø Trease and Evans Pharmacognosy
Ø C.K.Kokate, A.P.Purohit, S.B.Gokhale, Pharmacognosy
Ø Materia medica.
Ø Fundamental of experimental pharmacology by M.N Ghosh.
RELATED LINKS:
Ø http://www.google.com
Ø http://www.Rxlist.com
Ø http://www.sciencedirect.com
Ø http://www.pubmed.com
Ø http://www.medline.com
Ø http://www.rguhs.ac.in
Ø http://www.wileyinterscience.com
Ø http://www.drugbank.com
7.2
7.3 Does the study require any investigation or interventions to be conductedon
Patients or other humans or animals?
YES, albino rats required for anti-inflammatory study
7.4 Has ethical clearance been obtained from your institution in case of 7.3?
The study is cleared from ethical committee of the institution
· Ref No.25/4/2007 – AWD, Reg No. 1135/a/07CPCSEA (Copy enclosed.)
ENCLOSURE-III
8. LIST OF REFERENCES:
1. Robbers JM, Speedie and Tyler V. Pharmacognosy and pharmacobiotechnology. Williams and Wilkins, Baltimore. 1996;1–14.
2. Qureshi R, Bhatti GR. Wild gourd-A green medicine. Medicinal and aromatic plant abstract 2008;30(1):122.
3. Mukherjee A, Parmar NS, Saraf S. National convention on current trend in the herbal drugs and annual conference of pharmacognosy. The National alternative, Gandhinagar, Gujarat, India. Jan 2003;12.
4. Balakrishnan BR, Sangameswaran B, Ahmed S, Bhaskar VH. Anti-hyperglycemic activity of roots of Tephrosia Villosa. Plant Archives. 2007;7(2):729-231.
5. Zainul Amiruddin Zakaria, Loo Yi Wen, Nurul Izzah, Abdul Rahman et al. Antinociceptive, Anti-Inflammatory and Anti-pyretic Properties of the Aqueous Extract of Bauhinia purpurea Leaves in Experimental Animals. Medical Principles and practice. 2007;16:443–449.
6. Iwu MW, Duncan AR, Okunji CO. New antimicrobials of plant origin. Perspectives on new crops and new uses. ASHS Press, 1999;457–462.
7. Rani P, Khullar N. Anti microbial evaluation of some medicinal plants for their anti-enteric potential agent multi drug resistant salmonella typhi. Phytother.res. 2004;18(8):670.
8. Khouri NA, Akawi Z, Daradka H. Effect of short term treatment with Citrullus Colocynthis on the lipid profile and other blood biochemical parameters in albino rats. Asian Journal of Chemistry 2007;19(2):1468-1472.
9. Mythreye R, Murugan M, Muthysamy M, Venkatesh S, The anti-microbial activity of leaves of Bauhinia Tomentosa. Indian Journal of Pharmaceutical Science. 2005;67:732-734.
10. Pettit GR, Numata A, Iwamoto C, Usami Y et al. Isolation and structures of bauhiniastatins 1-4 from bauhinia purpurea. Journals of Natural products 2006;69(3):323-327.
11. Vijayan S, LalithaG, Kirti PB. Prokaryotic expression of a constitutively expressed Tephrosia villosa defensin and its potent anti-fungal activity. Applied Microbiology and Biotechnology. 2008;80(4):1023-1032.
12. Ghosh MN. Fundamentals of experimental pharmacology. 2005;3:190-192.
9 / Signature of Candidate
10 / Remarks of Guide / The Present Work Found To Lead New Dimention In The Field Of Pharmacetical Biotechnology
11 / Name and Designation of
11.1 Guide
11.2 Signature / Dr. T. TAMIZH MANI M.Pharm., Ph.D.,
Professor
Department of Pharmaceutical Biotechnology,
Bharathi College of Pharmacy,
Bharathi Nagara.
Karnataka-571422.
11.3 Co- Guide (if any)
11.4 Signature / Dr. GURUKAR MATHEW.S Ph.D.,
11.5 Head of Department
11.6 Signature / Dr.GURUKAR MATHEW.S Ph.D.,
Professor and Head,
Department of Pharmaceutical Biotechnology,
Bharathi College of Pharmacy,
Bharathi Nagara.
Karnataka-571422.
12 / 12.1 Remarks of the Chairman and
Principal
12.2 Signature / Recommended for approval
Prof. Dr. TAMIZH MANI. T Ph.D
Principal
Bharathi college of pharmacy
Bharathi Nagara
Karnataka-571422