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Pharmacognostical, Phytochemical and
Antipyretic activity studies on the roots of
Wattakaka volubilis (L.f.) Stapf.

SYNOPSIS FOR
M. PHARM DISSERTATION

SUBMITTED TO

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
BANGALORE, KARNATAKA
BY

AMIT KUMAR SHUKLA
I- M. PHARM (2008-2009) PHARMACOGNOSY
DEPARTMENT OF PHARMACOGNOSY
M.S. RAMAIAH COLLEGE OF PHARMACY
BANGALORE- 560054

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
BANGALORE, KARNATAKA


ANNEXURE – II

PROFORMA FOR REGISTRATION OF SUBJECT FOR
DISSERTATION



1.



NAME OF THE CANDIDATE AND ADDRESS

AMIT KUMAR SHUKLA,

94-H R/0 MUGALHAN, P/0 JHUNGIAN, GORAKHPUR,

UTTAR PRADESH, INDIA.





2.


NAME OF THE INSTITUTION

M.S. RAMAIAH COLLEGE OF PHARMACY, BANGALORE.





3.


COURSE OF STUDY AND SUBJECT

M.PHARM


PHARMACOGNOSY




4.


DATE OF ADMISSION

09.04.2008





5.



TITLE OF THE TOPIC

Pharmacognostical, Phytochemical and Antipyretic activity studies on the roots of Wattakaka volubilis.









6.

6.1

6.2

6.3


6.3.1

6.3.2

7.

7.1


7.2

7.2.1

7.3
7.4

8.

9.

11.

12.

BRIEF RESUME OF THE INTENDED WORK

NEED FOR STUDY

Nature always stands as a golden mark to exemplify the outstanding phenomenon of symbiosis. The biotic and abiotic elements of nature are all inter dependent. Plants are indispensable to human beings. The knowledge of drugs has accumulated over thousands of years as a result of man’s inquisitive nature which has provided today many effective means of health-care.


All phyla of plants “Of which concervative estimates place the total number of known spieces at approximately 335000” contains speices that yield official and unofficial products of medicinal importance. The oldes known herbal is Pen-t’sao written by emperor Shen Nung around 3000 B.C. It contains 365 drugs, one for each day of the year1.

‘Murva’ is an important ayurvedic drug used for the treatment of general body ailments like fever, laxative, urinary diseases, pruritus, sterlity, rigidity in lower limbs and skin diseases2. The botanical sources of Murva are Clematis tribola, Sanseviera zeylanica, Maerua oblongifolia, Chonemorpha macrophylla, , Helicteres isora. and Wattakaka volubilis3. The accepted botanical source is Marsdenia tenacissima4.

The genus Wattakaka belongs to the family Asclepiadaceae, It is known as cotton milk plant and green wax flower5, a tall, woody climber, with densely lenticellate and pustular branches. The roots contain a glucoside and traces of an alkaloid6. It is distributed in most parts of India7, Taiwan, Cambodia, Nepal, Sri Lanka8.

Several glycosides are found in Wattakaka volubilis such as dregeosides H, Dp1, Da1, Gp1,Ga1-isolated9. Glycosides-dregeosides Ap1, Ao1, Aa1, A11, C11, Kp1 and Ka1-isolated from stem10. Drevogenin D, mp.227 degree, isolated from seeds and characterized as 3β, 11β, 12β, 14β, 20 prntahydroxypregn-5-ene11.

Wattakaka volubilis is used in the treatment of kidney stones12; roots are reported to be a remedy for colic pain; plant found to have mild CNS-depressant, anthelmintic, antispasmodic, cytotoxic, antimutagenic and anticancer activities13.

The present investigation is aimed to carry out acute toxicity and antipyretic activity of roots of Wattakaka volubilis (L.f.) Stapf. Since “MURVA” is reported to be used against fever.



REVIEW OF LITERATURE


  • Three novel Polyoxypregnane glycosides, volubiloside A, B and C isolated from the flowers of Degea volubilis was reported14.



  • An unusual novel triterpenoid ether,multiflor-7-en-12,13-ether and a new multiflor-7-en-12α-ol from Wattakaka volubilis has been reported15.



  • Eight polyoxypregnane glycosides, Marsdenoside A-H, isolated from the CHCl3-soluble frcation of the ethanolic extract of the stem of Marsdenia tenacissima, along with six known glycosides and two known Polyoxypregnane aglycones has been reported16.



  • Two oligosaccharides from Marsdenia roylei, and their structures determined as O-β-D-oleandropyranosyl-(1→4)-O-β-D-digitoxopyranosyl -(1→4)-cymaral and ethyl O-β-D-oleandropyranosyl-(1→4)-O-3-O- methyl -6-deoxy- β-D-allopyranoside, respectively has been reported17.





  • Novel saponins hainaneosides A & B isolated from Marsdenia hainanensis, these saponins possesing fertility- regulating activity has been reported19.



  • A new triterpenoides named griffithol with oleanic acid, longispinogenin and chichipegenin isolated from the stem of Marsdenia griffithii was reported20.



  • A pregnane glycosides ester was isolated from Marsdenia koi by column

chromatography. The structure was identified on the basis of IR, 1H-

NMR, 13C- NMR and FAB-MS has been reported 21.





  • The effect of aqueous and solvent extracts of Marsdenia volubilis on growth of human pathogenic bacteria and fungi was studied in laboratory condition. Plant possess a wide range antibacterial and antifungal effect has been reported 22.



  • Antidiabetic properties of plants (Androgrphis lineate, Androgrphis paniculata, Costus speciosus Wattakaka volubilis) used by two major tribal groups in South Tamilnadu, India has been reported 23.



  • Protection against Selenite Cataract in rat lens by Drevogenin D, a triterpenoid Aglucone from Dregea volubilis was reported, treatment with Drevogenin D at a concentration of 50µg/ml24.



  • Hypoglycemic activity of Marsdenia volubilis was reported. The inhibitory effect of aqueous extracts of Eugenia jambolana seed, Marsdenia volubilis leaves and Phylianthus niruri on glucose blood levels was investigated in fasting, normal ,alloxan induced diabetic25.



  • Apply leaf juice of Wattakaka volubilis by adding little lime on affected part to cure sprains once day till cured was reported26.



  • Aliquots of extracts from ethnopharmacological plants that have activity against the effects of sarafotoxins present in snake venom are isolated from Wattakaka volubilis was reported27.


AIMS AND OBJECTIVES OF THE STUDY


AIMS
1. To undertake Pharmacognostical investigation on roots of Wattakaka volubilis.
2. To investigate and evolve Phytochemical parameters for the drug.

3. To investigate and evaluate acute toxicity and antipyretic activity of the drug.



OBJECTIVES



  1. To study taxonomical characters of the plant which help in the investigation.




  1. To understand the macro and microscopical characters of the drug besides phytochemical analysis. This helps in evolving diagnostic characters for the identification of the drug and also contribute towards Pharmacopeial standards.




  1. To carry out acute toxicity studies.




  1. To evaluate the antipyretic activity of the drug.

MATERIALS AND METHODS

SOURCE OF DATA



  1. Literature survey, Websites.



  1. Journals and Publications.




  1. Lab based studies.

METHOD OF COLLECTION OF DATA

Field and laboratory based studies
Field Work:

The selected plant roots will be collected from forests. The voucher specimen will be collected and deposited at the herbarium/museum of the department.


Laboratory work:



  1. Taxonomical studies:

The plant material will be identified by using various Floras like the Flora of Coorg28, Flora of Presidency of Bombay29, Herbarium specimens will be prepared following the method of Jain and Rao as per International standards30.




  1. Pharmacognostical studies:

Free hand sections of the drug will be taken following Wallis31. Microscopical investigation and histochemical tests will be carried out following Evans32.

Quantitative microscopy for carrying out the measurement of tissues will

be done by using stage and ocular micrometers. Photomicrographs will be taken and images captured on computer.




  1. Phytochemical studies:

Physical constants of drug will be determined as per Indian Pharmacopoeia33, and Phytochemical tests for detection of organic constituents will be done as per Harborne34. The chromatographic, fluorescence and HPTLC

studies will be carried out following Krebs et al35, Chase and Pratt36.



  1. Pharmacological studies:


Acute toxicity studies:
Determination of minimum lethal dose of the drug extracts will be performed followed by OECD guidelines37. Swiss albino mice of either sex of weight range of 20-25g will be used for acute toxicity studies. Mice which do not fall within this weight range will not be included in the study38.
50 mice will be used for determination of minimal lethal dose studies.
Antipyretic activity :39 40 41

Antipyretic activity of the drug extracts will be studied on albino rats

(Wistar strain) of either sex in the weight range of 170-190 g. Animals outside this weight range will be excluded from the study.

No. of groups = 6 (each)

(each containing 6 rats)



  • Group 1: vehicle control group




  • Group 2: standard group, 45mg /kg of Paracetamol p.o.




  • Groups 3 & 4: Two doses each of aqueous extract of drug p.o.




  • Groups 5 & 6: Two doses each of alcohol extract of drug p.o.




  • Total: 36 rats will be required.

Basal body temperature of all animals will be noted first. Fever will be induced in rats by subcutaneous injection of Brewer’s yeast (20%w/v) in distilled

water. After 18 hours rats will show increased body temperature. Drugs will be administered to febrile rats once and body temperature will be noted at specific

intervals of (4 hrs.) after drug treatment.


The results will be subjected to statistical analysis by one way ANOVA followed by Tukey-Kramer multiple comparison test.


Does the study require any investigation to be conducted on patients or other humans or animals? If so, please describe briefly.
Yes, Animal experiments on rats will be required for studying antipyretic activity and mice will be required for acute toxicity studies.
Has ethical clearance been obtained from your institution in case of

7.3?
Yes, ethical committee clearance has been obtained. Certificate copy enclosed

LIST OF REFERENCES
1. Kokate CK, Purohit AP, Gokhale SB. Pharmacognosy, 26th edition, Nirali

Prakashan, Pune. 2004:1-5.


2. Kolammal M. Pharmacognosy of Ayurvedic Drugs, series-1

Trivandrum, Ayurveda research institute. 1978:1.


3. Sharma PV. Drvyaguna vignan vol-5, 3rd edition, Chaukhambha bharati

Academy, Varanasi. 2006:232.


4. Anonymous. The Ayurvedic Formulary of India Part-1,1st edition,

Government of India Ministry of health and Family Planning. 1978:257.

5. Gurudeva MR. Botanical and Vernacular Names of South Indian Plants,

Bangalore Divyachandra pakashana. 2001: 419.


6. Anonymous. The Wealth of India. Raw material, vol-10, New Delhi: CSIR.

1998: 512.


7. Chatterjee A, Parkrashi SC.The Treatise on Indian Medicinal Plants, vol-4,

NISCAIR, New Delhi. 1992:131.




  1. Hooker BJD, Brit Fl, FOC 1883; 16:250.

9. Rastogi RP, Mehrotra BN. Compendium of Indian Medicinal plants,

vol-4, Lucknow CDRI and New Delhi NISC. 1985:763.

10. Rastogi RP, Mehrotra BN. Compendium of Indian Medicinal plants,

vol-3, Lucknow CDRI and New Delhi NISC. (1980-1984):678-79.
11. Rastogi RP, Mehrotra BN. Compendium of Indian Medicinal plants,

vol-1, Lucknow CDRI and New Delhi NISC. (1960-1969):433-43.


12. Singh VK, Govil JN. Recent progress in medicinal plants ethnomedicine

and pharmacognosy vol-1, studium press LLC. 2002:231.


13. Levekar GS. Data Base on Medicinal plants used in Ayurveda & Siddha

vol-8, Central council for research in Ayurveda & Siddha, New Delhi.2007:

272-78.
14. Shau NP, Panda N, Mandal NB, Banerjee S, Koike K, NikaidoT.

Polyoxypregnane glycosides from the flowers of Dregea volubilis.



Phytochemistry 2002; 61:383-88.


  1. Niranjan RVL, Ravikanth V, Vijendra RA, Prabhakar RT, Venkateswarlu Y.

An unusual novel triterpenoid ether, multiflor-7-en-12,13- ether and a new

multiflor-7-en-12α-ol from Wattakaka volubilis. Tetrahedron Letter 2002;

43:1307-11.



  1. Deng J, Liao Z, Chen D. Marsdenoside A-H, polyoxypregnane

glycosides from Marsdenia tenacissina. Phytochemistry 2005;66:1040-

51.
17. Kumar A, Khare A, Khare NK. Two oligosaccharides from Marsdenia-



Roylei. Phytochem 1999;50:1353-57.
18. Shen X, Hu YJ, Xu J, Chen HD, Shen YM. Studies on the chemical

constituents of Dregea sinensis Hemsl . Acta Pharmaceutical sinica 1996

Aug; 31: 613-16.

19. Ma B, Fang T, Ma K, Ni J, Wu H, Ding WP, Jiang C. Novel saponins

hainaneosides A & B isolated from Marsdenia hainanensis. J Natu Product 1997 Feb;20:134-38.
20. Min H, Jijun C, Jun Z. A new triterpenoides from Marsdenia Griffithi.

Acta Botanica Yunnanica 1992 Mar;14:323-27.

21. Ruan JL, Wan YM, Chem GX, Ding WP. Structure determination of

Marsdekoiside C. A Pharma sinica 1991Sep;26:667-71.
22. Vijayal T, Devamma NM, Nirmala C. Antibacterial and antifungal activity

of Marsdenia volubilis in invitro conditions. International seminar on



medicinal plants 2008 Mar.
23. Ayyanar M, Sankarasivaraman K, Ignacimuthu S. Traditional herbal

medicine used for the treatment of diabetes. Ethnobotanical Leaflets 2008;

12: 267-80.
24. Biju PG, Gayatri DV, Lija Y, Abraham A. Protection against Selenite

Cataract in rat lens by Drevogenin D, a triterpenoid Aglucone

from Dregea volubilis. J Med Food 2007 Feb;10:308-15.

25. Jasmine R, Daisy P. Comparative effect of plant extracts on the

Bloodglucose levels of alloxan diabetic rabbits. J Exp Zoology 2004

Feb;7:337-40.


26. Sanyasi RML, Verma YNR, Kumar V. Apply leaf juice by adding little

lime on affected part to cure sprains once day till cured. Ethnobotanical



Leaflets 2008;12:217-26.
27. Subbiah V. Method for screening for endothelin-receptor antagonist activity

and for treating conditions caused by endothelin. United states Patent

Application Publication 0008710 A1 Jan 13, 2005.

28. Keshava MKR, Yoganarasimhan SN. The flora of Coorg, Vimsat

Publication, 1990: 284-85.

29. Cooke T. Flora of the presidency of Bombay, vol-2, DehraDun Bishan

Singh Mahendra Pal Singh. 2006:164-67.
30. Jain SK, Rao RR. Field and herbarium methods, New Delhi Today and

Tomorrow Printers and Publishers. 1976.


31. Wallis TE. Textbook of Pharmacognosy, New Delhi, CBS Publishers

and Distributors. 1985:355-58.


32. Evans WC. Trease and Evans Pharmacognosy, 15th edition, London

Saunders. 2002:137.


33. Anonymous. Indian Pharmacopeia, vol-2. New Delhi: Controller of

Publications. 1996; A-89:53-54.


34. Harborne JB. Phytochemical methods, A Guide to modern techniques

of plant analysis, 3 rd edition, London Chapman and Hall.1998: 278.


35. Krebs KG, Heunsen D, Wimmer H. Thin layer chromatography,

A laboratory hand book, 2 nd edition, London ELBS, 1969:204-55,855-09.

36. Chase CR, Pratt R. Fluroscence of Powdered vegetable drugs with

particular reference to the development of system of identifications

2 nd edition, J. Am. Pharm Assoc 1949:324-31.
37. OECD/OCDE, OECD guidelines for testing of Chemical-420, 2001 Dec.
38. Kulkarni SK. Hand book of experimental pharmacology, 3 rd edition, Delhi

Vallabh Prakashan. 1999:168-71.


39. Chatterjee TK, Mishra M, Pramanik KC , Bandyopadhyay D. Evaluati-

on of Anti-inflammatory, Antipyretic and Analgesic properties of



Biophytum sensitivum (L) DC. Indian drugs 2008 Feb; 45:123-31.

40. Khan A, Haque E, Mukhlesur RM, Mossaddik A, Abdul AAl-B M,

Rahman M. Antipyretic activity of Rhizome of Drynaria querifolia in

rabbit. Pharma Biology 2007Apr; 45:312-15.

41. Ray D, Sharatchandra KH, Thokchom IS. Antipyretic, Antidiarrhoeal,

Hypoglycaemic and hepatoprotective activities of ethyl acetate extract of



Acacia catechu Willd. In albino rats. Indian J pharmacol 2006 Dec;38: 408-

13.



Signature of the Candidate:

Remarks of the Guide:

Name and Designation:

11.1) Guide : Dr. V. Madhavan. M. Pharm, Ph.D.

Principal, Professor & Head

Department of Pharmacognosy

M. S. Ramaiah College of Pharmacy

Bangalore.

11.2) Signature:

11.3) Co-Guide: Not Applicable

11.4) Signature: Not Applicable

11.5) Head of the Department:

Dr. V. Madhavan. M. Pharm, Ph.D.

Principal, Professor & Head

Department of Pharmacognosy

M. S. Ramaiah College of Pharmacy

Bangalore.



11.6) Signature:

12.1) Remarks of the Principal:

12.2) Signature:




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