ESTIMATION OF AGNUSIDE, NEGUNDOSIDE &1,4 DIHYDROXY BENZOIC ACID FROM

Vitex negundo. Linn .

BY HPLC.

SYNOPSIS FOR

M.PHARM DISSERTATION

SUBMITTED TO

RAJIV GANDHI UNIVERSITY OF HEALTHSCIENCES

BY

Sona Marium Panicker

Department of Pharmacognosy

THE OXFORD COLLEGE OF PHARMACY

BANGALORE-68

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

KARNATAKA, BANGLORE.

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION.

1. / Name of the candidate and Address / Sona Marium Panicker
The oxford college of pharmacy ,
N6/9,1stcross,Begur road
Hongasandra,
Bangalore-68.
PERMANENT ADDRESS
D\0.M.I.Alexander panicker
Panickers,Thevally,
Kollam-691009,
Kerala.
2. / NAME OF THE INSTITUTION / The Oxford College of Pharmacy,
Begur road,
Hongsandra,
Bangalore-68
3. / COURSE OF STUDY AND SUBJECT / M.Pharm,Pharmacognosy.
4. / DATE OF ADDMISSION / 28thApril,2010
5. / TITLE OF THE TOPIC / ESTIMATION OF AGNUSIDE, NEGUNDOSIDE&1,4 DIHYDROXY BENZOIC ACID FROM
Vitex negundo.Linn.
BY HPLC.

6. Brief resume of intended work:

6.1-Need for study:

Vitexnegundo.Linn. is also called as nirgundi, nagod, nakkilu banna belongs to the Verbenaceae. It is distributed in outer region of Himalaya and some district of Himachal Pradesh1.

The leaves contains alkaloids and glycosides nirgundoside,nishindaside. The principle constituents of the leaf juice are casticin, isoorientin, chrysophenol D, luteolin, p–hydroxybenzoic acid and D-fructose. The main constituents of the oil are sabinene, linalool, terpinen-4-ol, β-caryophyllene, α-guaiene and globulol constituting 61.8% of the oil. The leaves contain fragrant, volatile oil and resins.2 The fruits contain resin, astringent organic acids, alkaloids and a pigment.The plant also contains flavanoids, reducing sugars, sterols, resin and tannins.3 From seedsn-triacontane, n-hentriacontane, n-pentatriacontane, n-nonacosane, sitosterol, p-hydroxybenzoic acid and 5- oxyisophthalic acid isolated and also 3, 4- dihydroxybenzoic acid is isolated from seeds. Vanillic acid, p-hydroxybenzoic acid and luteolin are isolated from bark. From essential oil pinene, limonene, camphene, phellandrene, methylheptanone, cumene, linalool, camphor, 4-terpineol, citral, caryophyllene oxide, caryophyllene, terpinyl acetate, benzaldehyde & cinnamaldehyde identified. Nishindaside and nirgundoside isolated from leaves.4

The plant has anti-inflammatory, antibacterial,antifungal and analgesicactivities. These properties are useful in the treatment of superficial bruises, injuries, sores and skin infections. The essential oil of Vitex negundo.Linn. has been used to reduce inflammation and swelling of joints due to rheumatism and injuries.5

A survey of literature on Vitex negundo.Linn.revealed pharmacological reports on the plants like analgesic activity,anticonvulsant activity,insecticidal and pesticidal,antioxidant properties,antihistamicactivity,antirheumatic,antiflatulent

Although a lot of work has been done on pharmacological activities and phytoconstituents isolation of bark but no work was done on the estimation by HPLC. Therefore the aim of present study is to carry out the estimation of agnuside , negundoside and 1,4 dihydroxy benzoic acid from vitexnegundo.Linn. by HPLC and finding an alternative methods for above.

6.2-Review of Literature.

1.Thin Layer chromatography method has been established for analysis of negundoside inleaf powder of vitexnegundo.Linn .The methanol extract of the leaf was used for experimental work. The validated TLC densitometer method can be used for routine quality–control analysis of Vitex negundo.Linn.leaf powder and for quantitative analysis of negundosides.6

2. The ethanolic extract of leaves of Vitex negundo.Linn.was found safe as LD50 dose (by oral route) of it was recorded in non toxic dose range. In this paper, medicinal activity and pharmacological activities of various parts of plant has been reviewed.7

3. The oral anti-inflammatory, analgesic and antihistamine properties of mature fresh leaves of Vitex negundo.Linn. (Verbenaceae) claimed in the Ayurvedic medicine by orally treating water extract of the leaves to rats. The leaves showed an inversely dose-dependent in vivo antihistamine and in vitro prostaglandin (PG) synthesis inhibition, membrane stabilising and antioxidant activities.8

4.Hepatoprotective activity of Vitex negundo.Linn. leaf ethanolic extract was investigated against hepatotoxicity produced by administering a combination of three anti-tubercular drugs isoniazid -7.5mg/kg, rifampin -10mg/kg and pyrazinamide -35mg/kg for 35days by oral route in rats.9

5.The antinociceptive activity of ethanolic leaf extract of Vitex-negundo.Linn. was shredded by tail flick test in rats and acetic acid induced writhing in mice.10

6. The ethanol extract of vitex negundo.Linn. was found to possess potentanalgesiceffect which is most likely to be mediated by its anti-inflammatory activity rather than through opioid receptor system and therefore could partially explain the anti-nociceptive effects of Vitexnegundo.Linn.11

7.The larvicidal activity was assessed by the procedure of WHO with some modification. The highest larval mortality was found in leaf acetone extract of Adhatodavasica, ethyl acetate of bark ofAnnonasquamosa, methanol leaf extract and flower of Cassiaauriculata, l ethyl acetate of leaf of Hydrocotylejavanica, methanol leaf extract and seed of Solanumtorvum and leaf hexane extracts of Vitexnegundo.Linn. against the fourth instar larvae of Anophelesstephensi and Culex quinquefasciatus.12

9. The cardiotonic effect of aqueous extract of leaves of VitexnegundoLinn. was studied by using isolated frog heart perfusion technique hence, as compared to standard, test drug showed wide therapeutic index. Compared to standard.13

10. Theanti-inflammatory activity ofZingiber officinaleRosc, Vitex negundoLinn and Tinospora cordifolia (wild) Miers in albino rats were studied by acute and subacute inflammation.14

11. The review on vitex negundo.Linn, highlights the various potential therapeutic feature, which can be used in the drug designing, process to find solution for the unsolved infectious and non infectious deformities without side effect.15

6.3Objective of study:

The objective of present study is to estimate active constituents of Vitex negundo.Linn
viz agnuside,negundoside &1,4 dihydroxy benzoic acid by HPLC and to validate the method as per ICH guidelines.

7.Materials and Methods:

7.1 a)Source of data :

  • Medline,
  • RGUHS- digital library
  • Other Journals from The Oxford College of Pharmacy and Natural Remedies Pvt.Ltd.Bangalore.

b) Place of work:

Natural remedies Pvt. Ltd. and The Oxford College of Pharmacy, Bangalore.

7.Material and Methods

7.2 Method of collection of data:

  • Collection- Authenticated sample ofVitex negundo will be collected from Natural remedies Pvt. Ltd. Bangalore.
  • Extraction- Extracts of drugs will be prepared using different solvents by maceration/reflux.
  • Estimation-method development, validation as per ICH guidelines and SOP.

7.3Does the study require any investigations to be conducted on patients or other human being or animals?

If so, please describe briefly.

-Not applicable-

7.4 Has ethical clearance been obtained from the institution in case of 7.3?

-Not applicable-

8.Bibliography:

  1. Gupta AK, Neeraj T, Madhu S. Quality Standards of Indian Medicinal Plants. New Delhi.Indian Council of Medicinal Research2005; (3):358-66.
  2. Dr.Vinod D.Rangari Pharmacognosy and Phytochemistry–II Career publishation. 2003; (2):299-302.
  3. A.K Nadkarni.Indian Materia Medica, Bombay Popular Prakashan Private Limited, Mumbai. 2000; (1):1278-80.
  4. Khare CP.Encylopedia of Indian Medicinal Plants,Springer. 2004:474-476
  5. Saharan,Mond, Chouhan, Gupta.Text book of Pharmacognosy student edition;2008:382-384
  6. Prashant J, lokhande, Jayant k.Verma.Quantification of negundoside in vitexnegundo leaf by HPTLC,Journal of Planar Chromatography. 2009 March; (22):225-228
  7. vishal R, Tandon. Database on Medicinal and Biological activity of vitexnegundo, Natural Radiance. 2005May–june; 4(3):162-165.
  8. Dharmasiri MG, Jayakody G. Galhena, Liyanage SSP, Ratnasooriya WD,Anti-inflammatory and Analgesic activities of mature fresh leaves of Vitex negundoJournal of Ethnopharmacology. August 2003:199-206
  9. Vishal R. Tandon V. Khajuria B. Kapoor D. Kour S. Gupta.Hepatoprotective activity of Vitex negundo leaf extract against anti-tubercular drugs induced hepatotoxicityFitoterapia, 2008Dec79(7-8):533-53.
  10. Gupta RK and Tandon VR. Antinociceptive activity of vitexnegundo leaf extract.Indian journal physiol pharmacol. 2005;49(2):163-170.
  11. Zheng CJ,Tang WZ, Huang BH, Han T.Bioactivity guided fractionation for analgesic properties and constituents of Vitex negundo L. seeds.Phytomedicine. 2009 June:16(6-7):560-567.

12.Kamaraj c, Abdul Rahman A, Bagavan A etal. Larvicidal efficacy of 13 medicinal extract against Anophelesstephensi and Culexquinquefasciatus (Diptera: Culicidae Unit of Bioactive Natural Products, Post Graduate and Research, Trop Biomed 2010 Aug; 27 (2): 211-19.

13. Pai PT, Adnaik RS, Mule SN, Naikwade NS, CS Magdum. Evaluation of cardiotonic activity of leaves of vitexnegundo.International Journal of Green Pharmacy. 2009Feb; 3( 4):306-309

14.Jana Utpalendu,Chattopadhyay RN, Prasad Shaw Badri.Preliminary studies on anti-inflammatory activity of Zingiber officinaleRosc.,Vitex negundoLinn and Tinospora cordifolia (wild) Miers in albino rat.Indian Journal of pharmacology. 1999June;31(3):232-233.

15. Rameshbabu Arun Prabhu, Anand Prem Rajan. Review on the therapeutic potential ofVitex negundo Linn. Journal of Pharmacy Research.2010 Aug; 3(8):1920-22.

9. / Signature of candidate
10. / Remark of guide / Recommended
11. / 11.1 Name and Designationguide / Dr. S. Kambhoja
Professor
Department of pharmacognosy.
11.2 Signature
11.3 Co – guide / B.Murali.
Senior Manger-Analytical chemistry(R & D)
Natural Remedies, Veerasandra Indl Area, Bangalore.
11.4 SIGNATURE
11.5 Head of the department / Dr. Padmaa M. Paarakh
Professor and Head
Department of Pharmacognosy
11.6 Signature
12 / 12.1 Remark of chairman and principal / Forwarded to university for scrutiny.
12.2 signature / Dr. Padmaa M. Paarakh
Principal
The Oxford college of Pharmacy,
Hongasandra, Bangalore-68.