EXPLORATION OF THREE SPECIES
OF TRADITIONALLY USED MEDICINAL PLANTS FOR
THEIR PHYTOCHEMICAL AND CYTOTOXIC SCREENING
SYNOPSIS FOR
M.PHARM DISSERTATION
SUBMITTED TO THE
RAJIV GANDHI UNIVERSITY OF HEALTH
SCIENCES, KARNATAKA
BY
MOHAMMED JAHANGIR PASHA.M
I M.PHARM (Mid-Stream)
DEPARTMENT OF PHARMACOGNOSY
VISVESWARAPURA INSTITUTE OF
PHARMACEUTICAL SCIENCES
BANGALORE-560070
(2011-2012)
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
KARNATAKA
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / Name of the candidate and address(In block letters) / MOHAMMED JAHANGIR PASHA.M
S/o MOHAMMED MUSTAFA
NIDAGHATTA V & P
MADDUR TQ, MANDYA DIST.
KARNATAKA STATE
PIN 571433
2. /
Name of the Institution
/VISVESWARAPURA INSTITUTE OF PHARMACEUTICAL SCIENCES
BSK- II STAGE, BANGALORE-560070
3. /Course of Study and Subject
/MASTER OF PHARMACY IN
PHARMACOGNOSY
4. /Date of admission to the course
/ 20.12.20105. / Title of the topic:
EXPLORATION OF THREE SPECIES
OF TRADITIONALLY USED MEDICINAL PLANTS FOR
THEIR PHYTOCHEMICAL AND CYTOTOXIC SCREENING
6
7
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/
BRIEF RESUME OF THE INTENDED WORK
6.1 NEED FOR THE STUDYIndia is blessed with rich plants biodiversity of more than 10,000 native medicinal plants. It is also boasts a long tradition of medicinal use of plants, it is estimated that at least 200 species are used by Indians for the treatment of cancer. Western Ghats of India is one area rich in biological diversity and traditional knowledge. Now attention is given for search of new anticancer agents because cytotoxic therapy has shown limitation of its possibilities and low efficiency in treatment of cancer. Cancer is second largest single cause of death, claiming over six million lives each year worldwide. Depending on the stage of cancer, surgery, radio therapy and chemotherapy are the most commonly used treatment modalities1.
In the present scenario the search for newer molecules is changing towards the natural bioactive substances owing to their fewer side effects and low cost. About 60% of currently existing anticancer drugs are derived from natural sources2.
Survey of literature reveals that various parts like roots, leaves, bark and flowers of plants are found to be used in ayurvedic system of medicine for is different pharmacological activities, especially in cancer.3,4,5.
However, these species have not been significantly screened for cytotoxic activity, hence the present study is under taken for phytochemical investigation and cytotoxic screening for roots of Ficus heterophylla, roots of Artocarpus lakoocha and roots of Opuntia dillenii.
6.2 REVIEW OF LITERATURE
Ficus heterophylla L.f.6,7 family: Moraceae, is a shrub: leaves distichous, juvenile often pinnate, flushed orange figs distributed in moist valleys, along the streams of India. Root powder of the plant is one of the ingredients in ayurvedic formulation Trayamanadya Ghrita and is given to cure abdominal tumors.
To the best of author’s knowledge, the species of ficus is unexplored and there are no references found in the literature on both phytochemical study and pharmacological activity. This brought up need to investigate the plant for its phytochemical constituents and for cytotoxic screening.
Artocarpus lakoocha Roxb8,9 (syn: A.lacucha Buch. Ham) is member of family: Moraceae and it is called monkey jack. Artocarpus lakoocha is a tree widely grown in Khasi hills and Western ghats of India. The genus is of appreciable importance as a source of edible fruit and species reported to contain phenolic compounds including flavonoids, stilbinoids, jacalin and lectin10. The extracts and metabolites of the species have been reported to possess several useful bioactive compounds. Recently, additional data is available on exploration of compounds in various biological activities including antioxidant ,11antibacterial, antifungal, antitubercular12and antiviral.13
However, roots of Artocarpus lakoocha is one among the ingredients used in ayurvedic formulation Virhat Manakadi Gudica used to cure tumors, and it is not pharmacologically screened for its claim. Hence, in the present study author has undertaken to evaluate the same.
Opuntia dillenii14 family: Cactaceae is a xerophyte grown on hill slopes in dry sunny areas of India. It is commonly called kanthari. Opuntia dillenii is obviously very reputable species as indicated by a compilation of literature on its pharmacological property and phytoconstituents.15,16 Important pharmacological activities reported in literature includes antioxidant activity,16antiinflamatory17, antidiabetic18,19 etc.
The roots of plant has traditionally been used in treating cancer in ayurvedic system of medicine but has not been scientifically proved for its cytotoxic study and previous studies on the plant, had led to the isolation of some unusual flavonoids. Hence the author decided to re-examine the species for its cytotoxic screening and phytoconstituents.
6.3 OBJECTIVE OF THE STUDY
§ To conduct systemic chemical investigation of
-Roots of Ficus heterophylla.
-Roots of Artocarpus lakoocha.
-Roots of Opuntia dillennii.
§ Detailed screening of Roots of Ficus heterophylla, Roots of Artocarpus lakoocha and roots
of opuntia dillennii for its cytotoxic activity.
MATERIAL AND METHODS
7.1 SOURCE OF DATA
§ Helinet.
§ Web resources and experimental work.
§ Pub med.
§ Science direct.
7.2 METHOD OF COLLECTION OF DATA
The data will be generated by performing the experiments using Allium cepa L root tip cells model. The standard information is collected from various journals, standard books available in library, Indian Institute of Science library, RGUHS digital library and from various national and international standard websites.
§ www.sciencedirect.com
§ www.pubmed.com
§ www.google.com
§ www.ijp-online.com
§ www.rguhs.ac.in/j-gate@helinet
7.3 COLLCETION OF PLANT MATERIAL
Plant materials will be collected from local areas of Karnataka and western Ghats of Maharashtra and species will be authenticated by taxonomist and used for the study.
7.4 PHYTOCHEMICAL SCREENING
SOLVENT EXTRACTION
Air dried plant materials will be powdered in willy mill and will be extracted in Soxhlet assembly. Each extract will be concentrated by distilling off the solvent and then evaporated to dryness on water bath. The extract obtained with each solvent will be weighed. Its percentage yield will be calculated in terms of air dried weight of plant material. The colour and consistency of the extract will be noted.
The obtained extract will be subjected to different phytochemical parameters.
Chemical investigation
- Qualitative chemical tests
- Column chromatography
- Spectroscopic analysis.
7.5 CYTOTOXIC SCREENING
a) Initiation and fixing of onion root-tips20
Onion bulbs will be placed in coupling jar containing tap water to initiate rooting. The roots will be allowed to elongate 1-2 cm before treatment. After attaining the normal size, the onion bulbs will be transferred to a petridish filled with different grades of concentration ranging from 2mg, 4mg, 6mg, 8mg (grades will be prepared by testing the solubility of plant extract with chemicals) and one of the Petri dish containing water without extract will be served as control. All these bulbs will be placed in an incubator at 80C for 3 hours.
b) Cytological studies/ Plant cytotoxicity analysis20
The extracts will be subjected to plant cytotoxicity analysis. The root tips of treated onion bulbs will be cut off and fixed in Carnoy’s fluid and transferred in 70 % alcohol. For cytological studies, the fixed roots will be rinsed and hydrolysed in 1N HCl for 5-8 min in oven at 600C. Squashes will be made in 2% Acetocarmine stain. Cytologically important slides will be made permanent with N-Butanol-acetic acid series. Observations will be made from 15 slides of each treatment and control cells will be examined for structural alterations and mitotic index. Photomicrographs of normal and aberrant diving cells will be taken in image capturing PC based microscope.
7.6 PARAMETERS TO BE EVALUATED
1) Phytochemical screening
a. Authentication of plants by taxonomist
b. Collection of plant material
c. Identification of phytoconstituents by preliminary investigation
d. Isolation of bioactive molecules
e. Identification of isolated compounds.
2) Cytotoxic Screening
a. Initiation and fixing of onion root-tips
b. Cytological studies/ Plant cytotoxicity analysis
7.7 STATISTICAL ANALYSIS
The result of cytotoxic activity will be statistically analyzed with the help of various biostatistical methods.
7.8 Does the study require any investigation or intervention to be conducted on patients or other humans or animals? if so, please mention briefly.
-NO-
REFERENCES
1. Durairaj AK, Vaiyapuri TS, Mazumder UP and Gupta M. Antineoplastic and antioxidant activities of Oxystelma esculentum on Swiss albino mice bearing Ehrlich’s ascites carcinoma. Pharmaceutical Biology 2009; 47:195-202.
2. Newman DJ, Cragg GM, Snader KM. Natural products as a source of new drugs over the period of 1981-2002. J Nat Prod 2003; 66:1022-37.
3. SG Nagendranath. The Ayurvedic System of Medicine. Calcutta: Kevalram Chatterjee; 1984. Vol 2 p. 171.
4. SG Nagendranath. The Ayurvedic System of Medicine. Calcutta: Kevalram Chatterjee; 1984. Vol 2 p. 522-23.
5. Kirtikar KR, Basu BD. Indian Medicinal Plants. 2nd ed.Allahabad: Lalit Mohan Basu: 1989. Vol 2 p.1177-78.
6. Kumar A , Bajpai O, Mishra AK, et al. Assessment of diversity in the genus Ficus L.(Moraceae) of Katerniaghat Wildlife Sanctuary, Uttarpradesh ,India. AJPS, 2011; 2:78-92.
7. Khare CP. Indian medicinal plants. Springer: 2007.Vol 1 p.266.
8. Shailendra kumar MB, Rakesh kumar MC, Bharat AC. Screening of selected biological activities of Artocarpus lakoocha Roxb. Journal of basic and clinical pharmacy 2010; 4:239-245.
9. Jagtap UB, Bapat VA. Artocarpus: A review of its traditional uses, phytochemistry and pharmacology. Journal of Ethnopharmacology 2010; 129:142-146.
10. Hakim EH, Achmad SA, Juliwaty LD,Makmur L.Prenylated flavonoids and related compounds of Artocarpus. Journal of natural medicine 2006; 60:161-184.
11. Supawatchara S, Donrawee L, Chaiyavat C. Antioxidant and toxicity activities of Artocarpus lakoocha Roxb. Heartwood extract. Journal of Medicinal Plant research 2010; 4(10):947-953.
12. Boonphong S, Baramee A, Kittakoop P Puangsombat P. Antitubercular and antiplasmodial prenylated flavones from roots of Artocarpus.Chiang Mai Journal of Science 2007; 34:339-344.
13. Likhitwitayawuid K, Sritularak B, Benchanak K, et al. Phenolics with antiviral activity from Millettia erythrocalyx and Artocarpus lakoocha. Nat Prod Res. 2005; 19(2): 177-182.
14. Hartmut B. Opunitia dillenii-An interesting and promising Cactaceae Taxon. J.PACD 2008;148-170.
15. Chen XP, Zhao X. Progress on Pharmacological action of Opunita dillenii study in China. Chin. J. Tradition.Med. Sci. Technol 1997;5: 335-336.
16. SuFeng C, Chiu LH, Gow CY.The protective effect of Opuntia dillenii Haw fruit against low-density lipoprotein Peroxidation and its active compounds. Food Chemistry 2008; 569-575.
17. Ahmed MS, Tanbouly ND, Islam WT, Saleem AA. Antiinflammatory flavonoids from Opuntia dilleni(ker-gawl) Haw.Phytother. Res. 2008;19: 807-809.
18. Perfumi M, Tocconi R. Antihyperglycemic effect of fresh Opuntia dillenii fruit.Intern.J. Pharmacogn. 1996; 34: 41-47.
19. Inas ZA, Abdallah. Evaluation of hypoglycemic activity of Opuntia dillenii Haw fruit juice in Streptozotocin induced diabetic rats. The Egyptian journal of Hospital Medicine 2008, 33;544-558.
20. Ghurde MU, Malode SN. Cytotoxic activity of Phyllanthus niruri L. whole plant extract on Allium cepa L. root tip cell. Int. J. Pharmacol. Biol. Sci. 2010; 4(4):107-114.
Anderson, R.L., C.L. Alden and J.A.
L., C.L. Alden and J. (1982) acid (NTA) on., 23
9 / SIGNATURE OF THE CANDIDATE
10. / REMARKS OF THE GUIDE
11. / NAME AND DESIGNATION
11.1 GUIDE / Dr. D.H.HARISH KUMAR
PRINCIPAL & PROFESSOR
DEPARTMENT OF PHARMACOGNOSY
V.I.P.S, BANGALORE-70.
11.2 SIGNATURE
11.3 CO-GUIDE (IF ANY) / Dr. RAJARAJESHWARI. N
ASST. PROFESSOR
DEPARTMENT OF PHARMACOGNOSY
V.I.P.S, BANGALORE-70
11.4 SIGNATURE / .
11.5 HEAD OF THE DEPARTMENT / Prof. RAMESH .C
HEAD OF THE DEPARTMENT
DEPARTMENT OF PHARMACOGNOSY
V.I.P.S, BANGALORE-70.
11.6 SIGNATURE
12. / 12.1 REMARKS OF THE
PRINCIPAL
12.2 SIGNATURE
1