“Phytochemical and Pharmacological Investigations on Uraria picta and its Substitutes”
SYNOPSIS FOR
M.PHARM. DISSERTATION
SUBMITTED TO
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
KARNATAKA
BY
UMESH B.MADAKATTI
I M.Pharm
Department of Pharmacognosy
AL-Ameen College Of Pharmacy
Bangalore – 560 027
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BANGALORE
ANNEXURE – II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / NAME OF THE CANDIDATE AND ADDRESS / UMESH B. MADAKATTI.At/Post -Ramdurg
Taluka- Ramdurg
Dist-Belgaum
Pin -591123
KARNATAKA
2. / NAME OF THE INSTITUTION / AL-AMEEN COLLEGE OF PHARMACY
HOSUR ROAD, BANGALORE – 560 027
3. / COURSE OF STUDY AND SUBJECT / M. PHARM. - PHARMACOGNOSY
4. / DATE OF ADMISSION / JUNE- 2009
5. / TITLE OF TOPIC : -
“Phytochemical and Pharmacological Investigations on Uraria picta and its Substitutes”
6.0 / BRIEF RESUME OF THE INTENDED WORK
6.1 / NEED FOR STUDY
Uraria picta DC. is a perennial herb widely distributed throughout India, Australia, Africa and all most all parts of Asia. It is one of the important constituents in ten herb formulation called ‘Dashmula, a well established Ayurvedic drug of the Indian system of medicine for treating general fatigue, oral sores and several gynecological disorders1.
Uraria picta is mainly used for its analgesic, anti inflammatory, antioxidant, aphrodisiacs and fracture healing properties. Different parts of the plants are reported for different activities such as, decoction of the entire plants are useful for cough, chill and fever etc., leaves are used as antiseptic, mainly for the treatment of gonorrhea, and also for acaricidal properties in human and animal2.
A recent study has demonstrated that the isoflavanones isolated from root bark of this plant have anti-microbial activity. Flavonoids are known to exhibit a range of biological activities including Anti-inflammatory, Anti-thrombotic, anti- viral, hepatoprotective and vasoactive properties due to their free radicals scavenging ability. Thus the major bioactive flavonoid rhoifolin has been considered as marker compound for qualitative and quantitative standardization of Uraria picta based on liquid chromatographic analysis1.
Now a days the requirement of Uraria picta is more for Ayurvedic preparations Because of thin roots, thousands of tons of the plant are needed for preparations in industries. In order to meet the rising demand for the raw drugs, adultration and substitution have become frequent which in turn results in compromised quality of herbal medicines. There are controversies with respect to the botanical identities of Uraria picta available in market and used by Ayurvedic manufacturing unit. Uraria logopodioies DC, Desmodium gangeticum(L) DC, Pseudarthria viscida wight & Arn have been used as substitutes for Uraria picta. However no there are no reports on their comparative analysis.
Hence there is need for comparative phytochemical & pharmacological, evaluation of Uraria picta and its substituents. In present work it is proposed to carry out phytochemical analysis of Uraria picta, Uraria logopodioies DC, Desmodium gangeticum(L) DC, Pseudarthria viscida wight & Arn and standardize them for marker compound and compare their analgesic and antimicrobial activity.
6.2 / REVIEW OF LITERATURE
Uraria picta Desv. (Syn. Doodia picta Roxb; Fam. Papilionaceae),a suffruticose sparingly branched perennial herb having a height of 0.9–1.8 m, is distributed throughout Bangladesh, India, Sri Lanka, Tropical Africa, Malay Islands and the Philippines3 . Traditionally, the plant is used as an antidote to the venom of a dangerous Indian snake, Echis carinata. Its leaves are a good antiseptic and are used against gonorrhoea. The fruits and pods are effective against oral sores in children and the roots used against cough, chills and fever. Uraria lagopoides has been reported for its analgesic and anti-inflammatory activity and Uraria critina for nitric oxide-scavenging and antioxidant effects1,2,4.
Four species of Uraria recrded in flora of Taiwan are revised in identity, morphology including Uraria picta and Uraria lagopodioides.5
Uraria species is reported to contain flavones, isoflavones, triterpenes and steroids.3,6
Two isoflavanones 5,7-dihydroxy-2’-methoxy-3’,4’-methylenedioxyisoflavanone, and 4’,5’-dihydroxy-2’,3’-dimethoxy-7-(5-hydroxychromen-7yl)-isoflavanone along 6 compounds including isoflavanones, triterpenes and steroids were isolated from roots of Uraria picta. The structures of these compounds where established unambiguously by UV, IR, MS and a series of 1D and 2D NMR anlysis3.
The plant is reported to be endangered in some parts of India so the in vitro micropropagation of Uraria picta by cotyledonary node and nodel explants has been reported 7. In vitro micropropagation of this plant has been reported using axillary bud culture and callus regeneration.8.
An isocratic RP-LC method was developed for the quantitation of rhoifolin in Uraria picta,. Rhoifolin was detected by UV detection at 265 nm and using C18 column. The method was validated and it was found to be reliable and specific1.
Aqueous and methanolic extracts of Uraria picta was evaluated for its Acaricidal properties in laboratories using human and domestic animal model. The total and fractionated extracts have been assessed for acaricidal activity on Lxodes ricinus. The results indicated that methanolic extract of this plant was more potent acaricide compaired to the aqueous extract9.
Comparative evaluation of Aqueous and methanolic extracts of roots of Asparagus racemosus and whole plants of Uraria picta were studied for anti-inflammatory activity using in-vitro and in-vivo animal models. The evaluation parameter of in- vitro model was NO radical scavenging assay and lipooxygenase assay and Carrageenan induced rat paw edema model was used in the in vitro model. Results indicated that the Uraria picta has better anti-inflammatory potential than Asparagus racemosus10.
Alcoholic and aqueous extract of aerial part of Uraria lagopoides was investigated for anti-inflammatory and analgesic activity using rat paw edema test at doses of 100 and 200 mg/kg body weight orally. Results showed that both the extracts at a dose of 100mg/kg body weight significantly(P<0.01) inhibited the edema compared with indomethacin and showed marked analgesic activity in mice ( P<0.01) compared with acetylsalicylic acid11.
Hypolipidaemic activity of Abana, an Indian herbomineral preparation containing Uraria picta as one of the ingredient was studied to investigate the action of Abana on the lipid and lipoprotein metabolism in albino rats. The results showed that chronic administration of Abana significantly reduce serum lipoprotein lipid components and apoprotein level in rats indicating Abana as cardioprotective and hypolidaemic agent12.
In another study Abana powder was administered orally in rabbits to investigate the effect of Abana . The results showed that administration of Abana for 3 days has an action similar to that chronic administration of isoprenaline which may be due to a specific depressant effect of Abana on the adrenergic receptor and a direct sensitization of the atrium13.
Different methods for testing analgesic activity by in-vivo methods using mice or rat have been reported including hot plate method, tail immersion method and acetic acid methods.15Analgesic activity of Uraria lagopoides has been reported using mice in acetic acid induced writhing test.9
Antimicrobial activity has been reported by using agar-plate techniques and microdilution titer technique.16 Antimicrobial activity of isoflavones isolated from Uraria picta has been reported against both Gram +Ve and Gram –Ve bacteria and fungi using cup late method and microdilution techniques.3
6.3 / OBJECTIVE OF STUDY
Aim: To carry out the Phytochemical investigates on Uraria picta and its substitutes and compare their analgesic and antimicrobial activity.
The objectives are:
1. Collection and authentification of whole Uraria picta, Uraria picta, Uraria logopodioies DC, Desmodium gangeticum(L) DC, Pseudarthria viscid wight Arn.
2. Pharmacognostical evaluation of all the selected plants.
3. Preparation and Phytochemical analysis of the extracts using HPLC and HPTLC.
4. Evaluation of antimicrobial and analgesic activity of selected extracts.
7.0 / MATERIALS AND METHODS
7.1 / SOURCES OF DATA
1. Information Centre, Al-Ameen College of Pharmacy, Bangalore.
2. Library, Centre for Pharmacognosy and Pharmaceutics, Foundation for Revitalisation of Local Health Traditions (FRLHT), Bangalore.
3. Search on MEDLINE, PUBMED, etc.
4. Google search.
5. Journals and publications.
7.2 /
METHOD OF COLLECTION OF DATA
PLACE OF STUDY:F.R.L.H.T, Bangalore
AL-AMEEN COLLEGE OF PHARMACY Bangalore.
1. Whole plant of Uraria picta ,Uraria logopodioies DC, Desmodium gangeticum Pseudarthria visdd. Wight & Arn. will be collected and authenticated at F.R.L.H.T.
2. Pharmacognostical study
The drug will be extracted successively with different solvents starting from
nonpolar to polar. All the extracts will be tested for presence of different
compounds.
The aqueous extracts of all the drugs will be prepared and subjected to TLC.
Amount of active Flavonoid rhoifolin will be estimated in the extracts by
HPLC and HPTLC methods.
3. Antimicrobial Study
All the aqueous extracts will be tested for antimicrobial activity against two
gram +ve and two gram –Ve organisum using cup plate method and microdilution titre
technique using 96 well plate.
4. Analgesic activity
The standadised aqueous extracts of the four Uraia species will be tested for analgesic activity
Animals -
Animal care and handling will be done according to the guidelines set by CPCSEA. Wister Rats of either sex weighing between 150-200 gms will be selected from inbred colony and maintained under standard control conditions. The animals will be fed with standard pelletized dit and will be provided free access to water ad. libitum.
Grouping –
Animals will be divided into 6 groups containing 6 animals in each groups
Group -1- Control group
Group -2- Standard group
Group -3-Test group (Sample 1)
Group -4- Test group (Sample 2)
Group -5- Test group (Sample 3)
Group -6- Test group (Sample 4)
Group 1 will receive distilled water solution.(1ml p.o)
Group 2 will receive Aspirin (100 mg/kg b/w) orally.
Group 3 to 6 will receive different extracts (500 mg/kg b/w) orally.
Method
Tail of the contrained animal will be individually dipped in beaker containing water at 580 ± 0.2 0C.Tail flicking respons will be taken at 30,60,&120 min after drug administration. The flicking response will be noted as an end point when the animal withdraw the tail from the beaker. Three basal reading will be taken. 180 sec. will be taken as cut off period. The data will be subjected to statistical evaluation.
7.3 /
PLACE OF WORK:
§ Al-Ameen College of Pharmacy, Bangalore.
§ Centre for Pharmacognosy and Pharmaceutics, FRLHT, Bangalore.
7.4 / Does the study require any investigations or interventions to be conducted on patient or other humans or animals? If so, please describe briefly.-Yes-
7.5 / Has ethical clearance been obtained from your institution in case of the above query?
Animal ethical clearance form is submitted in the college.
YES—Ref;NO.AACPI/M-108
8.0 / REFERENCES:-
1. Akhilesh KY, Deepti Y, Karuna S, Ram KV, Ajit KS, Madan MG Flavone glycoside Based Validated RP-LC Method for Quality Evaluation of Prishniparni (Uraria picta) Chromatographia 2009;69:653-658.
2. http://www.motherherb.com/index.html
3. Kirtikar KR, Basu BD, Indian Medicinal plants. 1993.
4. Mishra DN, Medicinal plant for treatment of fever in the MAdhavachikitsa tradition of medicine, Ind J of Trad know 2009; 8:352-361
5. Hiroyoshi O and Yu I, A revision of Uraria in Taiwan, 2007, Taiwania :52: 177-183
6. Rahman MM, Gibbons S, Alexander IG Isoflavanones from Uraria picta and their antimicrobial activity. Phytochemistry. 2007; 68:1692–1697.
7. Gaurav AM, Dhanorkar VM, Dhar BP, Lavekar GS In vitro propagation of medicial plant Uraria picta (Jacq.) Desv.ex DC. From cotyledonary node and nodal explants. Pharmacognosy Magazine. 2008;4(16):239-245.
8. Rao CS, Latha R, Josekutty PC, Balakrishna P, Anand A Micropropagation of Uraria picta, a medicinal plant, through axillary bud culture and callus regeneration.In Vitro Cellular & Development Biology 1998;34:2.
9. Lgboechi AC, Osazwa EO, Lawe UE Laboratory evaluation of the acaricidal properties of extracts from Uraria picta (Leguminosae).J Ethopharmacol, 1989;26(3):293-298.
10. Ahirrao P, Jagtap A, Shirke S, Fernandes B. Comparative assessment of anti-inflammatory potential of Asparagus racemosus and Uraria picta Life Sciences 2007 proc life sciences; Poster Communications :PC108
11. Hamid H, Abdullah S, Asif A, Alam M, Ansari SH Anti-inflammatory and analgesic activity of Uraria logopoides, Pharmaceutical Biology 2004;42(2):114-116.
12. Khanna AK, Chander R, Kapoor NK Hypolipidaemic Activity of Abana in Rats Fitoterapia; 1991; 62: (3): 271.
13. Pasnani JS, Hemavathi KG, Rajani AP Effect of Abana, An Ayurvedic preparation Rabbit Atrium and Intestine.J.Ethnopharmacology.1988;(24):287-302.
14. Drummond AJ, Waigh, RD, In: Pandalai, S.G, Recent Research Developments in Phytochemistry, Research Signpost, India, 2000. vol. 4; Pp.143-152.
15. Gerhard Vogel, Drug Discovery and Evaluation pharmacological assay’s, Second edition.
16. Michael J, Pelezar JR. Textbook of Microbiology. Fifth edition. Pp.504-505.
9. / SIGNATURE OF THE CANDIDATE
10. / REMARKS OF THE GUIDE / RECOMMENDED FOR RESEARCH AND SUBMISSION OF DISSERTATION
11. / NAME AND DESIGNATION OF THE GUIDE / Dr. SALMA KHANAM
Prof. & HOD
Department of Pharmacognosy,
Al-Ameen College of Pharmacy,
Bangalore.
11.1. / SIGNATURE
11.2 / CO-GUIDE / Dr. PADMA VENKAT
Joint Director, FRLHT,
Bangalore.
11.3 / SIGNATURE
11.4 / CO-GUIDE / Dr. Kshama Devi
Professor, Dept of pharmacology
11.5 / SIGNATURE
11.6 / HEAD OF THE DEPARTMENT / Dr. SALMA KHANAM
Prof. & HOD
Department of Pharmacognosy,
Al-Ameen College of Pharmacy,
Bangalore.
11.7 / SIGNATURE
11.8 / PRINCIPAL / Prof. B.G. SHIVANANDA
Al-Ameen College of Pharmacy,
Hosur road, Bangalore.
11.9 / REMARKS OF THE PRINCIPAL / RECOMMENDED FOR RESEARCH
12 / SIGNATURE