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Isolation of phytoconstituents and anti-oxidant activity studies on the seeds of linum usitatissimum Linn.” Synopsis for


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ISOLATION OF PHYTOCONSTITUENTS



AND ANTI-OXIDANT ACTIVITY STUDIES ON THE SEEDS OF

Linum usitatissimum Linn.
Synopsis for

M. Pharm Dissertation Submitted to



Rajiv Gandhi University of Health Sciences, Karnataka

Bangalore – 560041
ANOOP.T.V

Department of Pharmacognosy


The oxford college of pharmacy,

Bangalore -78
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

KARNATAKA, BANGALORE.


ANNEXURE-II




PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION






1

Name of the candidate and address



Mr. ANOOP.T.V


s/o Mr. VENUGOPAL.T,

“Indusree”, Opp:Aswini Hospital,

Ottapalam,

Palakkad, Kerala-679101





2


Name of the institution

The Oxford College of Pharmacy,

J.P Nagar,1st Phase,

Bangalore 560078



3


Course of study and subject

MASTER OF PHARMACY

(PHARMACOGNOSY)


4


Date of the admission

20th MAY 2008




5



Title of the topic:

“ISOLATION OF PHYTOCONSTITUENTS

AND ANTI-OXIDANT ACTIVITY STUDIES ON THE SEEDS OF

Linum usitatissimum Linn.







6



Brief resume of the intended work

6.1 Need for the study

The plant Linum usitatissimum Linn. belongs to the family Linaceae also called as Flax plant. It is distributed throughout India mainly in the Himalaya region.

The flax seed can be mainly subdivided into four types depending upon the color1.

The flax seed is reported to contain Lignin and linataine2. Some other like isolarciresinol, pinoresinol, mutaresinol, cyanogenic glycosides and some water soluble vitamins3. It contains cholesterol and campesterol4. The seed5 contain oil, mucilage and protein.

The fatty acid composition of total glycolipis6 ie Linolenic acid, Linoleic acid and Oleic acid were also isolated from different varieties of seeds .

Pharmacologically Linum usitatissimum Linn. is found to have Laxative7 and Anti-inflammatory effect8.The ability to reduce the hyper-cholesterol9 level of the seed have been reported. Seeds are found to have effect in the treatment of gonorrhea10.

The seed oil shows wound healing property11. Flax seed is found to

have anti-cancer effect12.

Although a lot of work has been carried out on the seed of the plant with respect to its pharmacological activities, Very less importance has been given towards the phytochemical investigation of the seeds of Linum usitatissimum Linn.



Therefore in the present study it is proposed to carry out isolation of the chemical constituents from of the seeds of Linum usitatissimum Linn. and in vitro anti-oxidant studies of the extracts / isolated compounds.







6.2 Review of literature

  • Based on the color,seeds are subdivided into white, yellow, red, black colored varieties1.Seeds are compressed, ellipsoid, smooth, shiny and are sweet, bitter, acrid.

  • Linatanine,a glutamic acid derivative and Lignans ie, Secoisolariresinol a diglucoside are isolated from seeds of the plant2.

  • Cyanogenic Glycosides3-linustatin and neo linustatin characterized as 2[(6-o- -D-Glucopyranosyl--D-glucopyranasyl)oxy]-2-methyl propanenitrile and(2R)[(6-o--D-glucopyranosyl --D- glucopyranasyl) oxy]2-methylbutanenitrile respectievely; carbohydrates,carotene,thiamine,riboflavin,niacin,pantothenicacid,5dehydroavenasterol,linusitamarin(phenyl propanoid glucoside),cyclolinopeptide B and tocopherols were reported from the plant Linum usitatissimum Linn.

  • Cholesterol(2),campesterol(26),stigmasterol(7),sitosterol(41).5-dehydro-avenastero(13).cycloartenol(9),and 24-methylenecycloartenol(2%) in seeds are charecterised by GC-MS4.

  • The seeds can be used as a high energy, high protein5 feed stuffs as well as an excellent source of dietary -linolenic acid for modifying plasma and tissue lipids.

  • The fatty acid composition of total glycolipids6 and its components are determined by GLC showed linoleic acid, linolenic acid, oleic acid as the major fatty acids. Composition of glycolipids from five varieties namely C-429, R-552, RLC-, RLC-6 and T-397 of Vidharbha region was investigated with chloroform: methanol (2:1)v/v7.

  • Flax seed has laxative7 effects arising from increased volume and consequent irritation of intestinal peristalsis from stimulation of stretch receptor.

  • The fixed oil (1,2,3 ml per kg) containing alpha linolenic acid obtained from the seeds of Linum usitatissimum Linn., were screened for their anti inflammatory8 activity using carrageenan, leukotriene and arachidonic acid induced paw edema models in rats and the anti inflammatory effects were compared with the standard drug Indomethacin, significant inhibition of paw edema was produced by the seed oil in the highest dose(3 ml per kg) in all the models.








  • Fifty-five mild to moderately hypercholesterolemic (≥5.1 to ≤9-8mmol/L) native of American post menopausal women were randomly assigned to control(A),flax seed(B),or flax seed + additional oat bran fiber(C) groups. Overnight fasting venous blood was collected at baseline and at the end of the treatment period, the analyzed lipid parameters indicated that dietary flax seed supplementation lowered total cholesterol9 and LDL cholesterol by approximately 7% and 10% respectively. HDL and triglyceride remained unaltered and no changes were observed in other clinical and hematological parameters.

  • Seeds are used internally for the treatment of gonorrhoea and used against the irritation of the genito-urinary tract10.

  • The flax seed oil mixed with lime water(Carron oil)has been a favorable application to burns and scalds11.

  • Low fiber intake and imbalance of omega 3 and omega 6 fats all contribute to excess cancer risk. Intake of Flax seed, an excellent source of dietary fiber, phyto estrogen, omega 3 fatty acid (-linolenic acid), and lignan will lower cancer risk12.


6.3 Objectives of the study:
The objective of the present study is to collect the seeds of Linum usitatissimum Linn. from Bayir chemicals, Bangalore and to carry out the extraction (maceration/ soxhlet method) and isolation of the phytoconstituent from selected extract based on phytochemical screening, and further characterization of isolated compound by spectral analysis( UV,IR and NMR spectroscopy). The different extracts and / or isolated compound would be studied for in vitro anti-oxidant activity by DPPH and Super oxide radical method.





7.1 Materials and methods:


  1. Sources of data




  • Chemical abstracts

  • Medline

  • Other Journal from the oxford college of Pharmacy Library

  • RGUHS digital library


b) Place of work

BAYIR CHEMICALS (Herbal Extracts Division), Bangalore.








7.2 Method of collection of data


  • Collection: - The seeds of Linum usitatissimum Linn. was collected and authenticated from Bayir Chemicals, Bangalore.

  • Extraction: - The extract of the drugs will be prepared using different solvent by maceration/ soxhlet.

  • Isolation: - Isolation of phytoconstituents will be done using column chromatography /fractionation method.

  • Identification and characterization:- The isolated compound will be identified using analytical methods like UV, IR, NMR spectroscopy.

  • Anti-oxidant activity: - The extract and/or isolated compound will be studied for in-vitro anti-oxidant activity by DPPH / Super oxide radical method.








7.3 Does the study require any investigation or interventions to be conducted on

Patients or other humans or animals?

Not applicable.








7.4 Has ethical clearance been obtained from your institution in case of 7.3?
Not applicable




8

Bibliography:


  1. Kulkarni P.H,Shahida Ansari.Indian Medicinal Science The Ayurvedic Plants.Sri Satguru Publications,Delhi.2004;Series No:132:201

  2. Khare CP.Encyclopedia of Indian Medicinal Plants Rational Western Therapy, Ayurvedic and Traditional uses,Botany.Springer,Verlag Berlin Heidelberg New York2004;289




  1. Sharma PC,Yelne MB,Dennis TJ,Joshy A,Prabhune YS,Borkar GB et al.Data base on medicinal plants used in Ayurveda.Central council for research in Ayurveda & Siddha New Delhi 2002;(4):14-16




  1. Rastogi Ram P ,Mehrotra BN,Shradha Sinha,Pant Pushpa,Seth Renu.Drug research Perspectives:A CDRI Series Compendium of Indian Medicinal Plants.Central Drug Research Institute,Lucknow and National Institute of Science Communication,New Delhi.1991;(2):416

  2. The Wealth of India: A Dictionary of Indian raw materials and Industrial Products. National Institute of Science communication and Information Resources council of scientific and Industrial Research, New Delhi.2003;(4):40-44




  1. Kulkarni AS,Khotpal RR,Bhakare HA.Glycolipids composition of some Indian Linseed varieties.Journal of Food Science and Technology.1998;35(3):245-246




  1. PDR for Herbal Medicines. Thomson PDR at Montvale.2004;(3):328




  1. Singh S,Nair V,Jain S,Gupta YK.Evaluation of anti-inflammatory activity of plant lipids containing alpha-linolenic acid.Indian J.Exp Biol.2008;Jun:46(6):453-6




  1. Anagha Patade, Latha Devareddy, Edralin A, Lucas, Kiranmagi Korlagunta, Bruce P. et al. Flax seed reduces total and LDL cholesterol concentrations in native American post menopausal women. Journal of Women’s Health. Mary Ann Liebert.Inc. 2008; (17)10:1089.




  1. Kirtikar KR, Basu BD.Indian Medicinal Plants.International Book distributors, Dehradun.2005;(1):408-409

  2. Drury CH.Ayurvedic useful plants of India with notices of their chief value in Medicine Commerce and art.Asiatic Publishing House,Delhi.2006;(2):278

  3. Donaldson Michael S.Nutrition and Cancer review of the evidence for an anti-cancer Diet. Nutrition Journal.Bio med central Ltd.2004;3:19








9


Signature of the candidate





10


Remarks of the guide:




Recommended


11






11.1 Name and Designation of

Guide

11.2 Signature



Mr.S.Kambhoja

Professor

Department of Pharmacognosy,

The Oxford College of Pharmacy, Bangalore-78










11.3 Co-Guide

11.4 Signature


Mr. Anjaneyalu

Vice-President

Herbal Extract Division,

Bayir Chemicals,Bangalore-91








11.5 Head of the department

11.6 Signature


Dr. Padmaa M Paarakh

Professor and HOD

Department of Pharmacognosy,

The Oxford College of Pharmacy,Bangalore-78



12




    1. Remarks of the Principal

Forwarded to the University for scrutiny






    1. Name of the Principal


12.3 Signature


Dr. Padmaa M Paarakh




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