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Rajiv Gandhi University of Health Sciences

Bangalore, Karnataka.
Preliminary Phytochemical Investigation and Screening of Diuretic Activity of Fruits Luffa acutangula ROXB””

A Protocol submitted to Rajiv Gandhi University of Health Sciences

Karnataka, Bangalore

in partial fulfillment of the requirement for the award of
MASTER OF PHARMACY

IN

PHARMACOGNOSY


RAVIKUMAR V SHEERI

Department of Pharmacognosy,

National College of Pharmacy,

Balraj-Urs Road,

Shimoga-577 201

Karnataka-INDIA.

A. BRIEF RESUME OF INTENDED WORK

A.1: Need for the study:
The plants are indispensable to man for his life. The history of herbal medicines is as old as human civilization. Even today mankind is obtaining drugs from plant source. A large number of plants are being used in Ayurvedic system of medicine, which is popular in our country. Herbs had been used by all cultures throughout history but India has one of the oldest, richest and most diverse cultural living traditions associated with the use of medicinal plants. In the present scenario, the demand for herbal products is growing exponentially throughout the world and major pharmaceutical companies are currently conducting extensive research on plant materials for their potential medicinal value1.

Luffa acutangula ROXB. is a large climber shrub, belonging to the family cucurbeataceae. It is found in India, Pakistan and Srilanka. It is evergreen climber found in all the season. Traditionally, the plant is used as a demulcent, diuretic, bitter tonic, nutritive and expectorant2. Fruit contains a bitter principle, luffeine. Seed contains a fixed oil of glycerides of palmitic, stearic and myristic acids. Fruit is considered demulcent, diuretic and nutritive. Seeds considered as purgative and emetic. A good source of calcium, iron and phosphorus3.

Number of studies are reported on pharmacological action from Luffa acutangula and various phytochemical constituents mainly luffangulin, a rhibosomal inactivating peptide have been isolated from seeds of Luffa acutangula4.

The claim of diuretic activity of Luffa acutangula has not been studied scientifically2. Hence the present study is aimed at investigation of diuretic activity of various extracts of Luffa acutangula and phytochemical investigation of biologically active extract.


A.2 Review of Literature:


  • A ribosome inactivating peptide, with an N-terminal sequence exhibiting pronounced similarity to that of the 6.5 kDa-arginine/glutamate-rich polypeptide from Luffa cylindrica seeds, was isolated from seeds of a closely related species, the ridge gourd Luffa acutangula4.

  • Seven oleanane-type triterpene saponins, acutosides A--G, were isolated from the herb of L.acutangula, and their structures were determined5.

  • An abortifacient protein, isolated from the seeds of L.acutangula has been estimated by SDS-polyacrylamide gel electrophoresis by using a procedure that involved acetone precipitation, ion exchange chromatography on CM Sepharose CL-6B and gel filtration on Sephadex G-506.

  • The dose-dependent CNS depressant activity on HPTLC pattern of ethanol extract of fruit of L.acutangula has been reported for the effect on behavioral changes, exploratory activity, barbiturate sleeping time, using appropriate standards in mice7.

  • Traditionally, the extracts prepared from both cold maceration and boiling the plant in the solvent under reflux were consumed by south-asian migrants in bradford, Yorkshire, Uk have been tested for their free radical scavenging activity (FRSA) in the DPPH (1,1-diphenyl-2-picrylhydrazilradical) screening assay, exhibiting

significant antioxidant activity of five vegetables, including L.acutangula8.

  • Luffaculin 1, a novel type of ribosome-inactivating protein (RIP) has been isolated from the seeds of L.acutangula and its protein sequence was done by X-ray sequence and crystal structure9.

  • The ethanolic and aqueous extract (100mg/kg) of L.acutangula have shown significant antihepatotoxic activity in druckrey rats by inducing hepatotoxicity with carbontetrachloride and paracetamol10.

  • Preliminary qualitative antifungal activity was carried out for eight plant extracts including L.acutangula on fungal pathogens, isolated from superficial mycotic infections against 143 clinically isolated strains of dermatophytes, 16 Candida spp and 25 Pityrosporum orbiculare11.

  • A chitooligosaccharide specific lectin (Luffa acutangula agglutinin) has been purified from the exudate of ridge gourd fruits by affinity chromatography on soybean agglutinin glycopeptides coupled to Sepharose-6B12.


A.3 Objective of the study:

1. Collection and authentication of L. acutangula

2. Drying of plant material

3. Preliminary extraction of drug using different solvents

4. Preliminary phytochemical investigation of extracts

5 Evaluation of diuretic activity of the extracts

6. Phytochemcial investigation of biologically active extract/s
B.MATERIALS AND METHODS:

B.1.Source of data:

The required data will be obtained from:



  1. Electronic data (Internet)

  2. Published research papers

  3. Review and research articles from journals

  4. Kuvempu university, library

  5. Library of IISc, National College of Pharmacy, Shivamogga

B.2. Methods of collection of data

  • The Plant will be identified and authenticated with the help of botanist

  • Drying of plant material

  • The plant of Luffa acutangula will be collected from local areas of Shivamogga, Karnataka

  • The plant material will be subjected to extraction with solvents of increasing polarity

  • Evaluation of Diuretic activity will be carried out by using Lipschitz test

B.3.Does the study requires any investigation to be conducted on patients or animals?

Wistar albino rats will be used for the investigations

B.4. Has ethical clearance been obtained from your institution?

Ethical clearance is provided by the institution.



Clearance number: NCP/IAEC/CL/15/12/2009-10.

C. LIST OF REFERENCES:

        1. Kokate CK, Text book of pharmacognosy Nirali prakashan New Delhi ed.4:3-4.

        2. Sahoo S, Ramesh DB, Rao YR, Debuta BK and Vibhuti Mishra N. Medicinal and Aromatic Plants Allied Publisher, New Delhi:364

        3. http://stuartxchange.com/patola.html.

        4. Hexiang Wang and Tzi Bun Ng. Luffangulin, a novel ribosome inactivating peptide from ridge gourd (Luffa acutangula) seeds. Life Sciences2002; 70(8):899-906.

        5. Nagao T, Tanaka R, Iwase Y, Hanazono H and Okabe H. Studies has been reported on the constituents of Luffa acutangula and Structures of acutosidesA--G, oleanane-type, triterpene saponins isolated from the herb.Chem Pharm Bull (Tokyo)1991;39(3):599-606.

        6. Yeung HW, Li WW and Ng TB. Isolation of a ribosome-inactivating and abortifacient protein from seeds of Luffa acutangula. International journal of peptide and protein research 1991;38(1):15-9.

        7. Misar AV, Upadhye AS and Mujumdar AM. CNS depressant activity of ethanol extract of Luffa acutangula var. amara C.B. Clarke fruits in mice. INIST-CNRS 2004; 66(4)463-465.

        8. Ansari NM, Houlihan L, Hussain B and Pieroni A. Antioxidant activity of five vegetables traditionally consumed by south-asian migrants in bradford, yorkshire, UK. Phytother. Res 2005; 19: 907–911.

        9. Xiaomin Hou, Minghuang Chen, Liqing Chen, Edward Meehan J, Jieming Xie and Mingdong Huang. X-ray sequence and crystal structure of luffaculin 1, a novel type ribosome-inactivating protein. BMC Structural Biology 2007; 7:29.

        10. Mukerjee A, Kaithwas G, Visen PKS Y and Saraf SA. Phytopharmacological screening of Luffa acutangula fruits for its antihepatotoxic activity. Ars Pharm 2007; 48 (4): 351-360.

        11. Kothavade RJ; Dabke NM and Chanderkar NG. Antifungal activity of some herbal extracts on fungal pathogens isolated from superficial mycotic infections. The Indian Practitioner 1997; 50(1); 21-24.

        12. Anantharam, Vellareddy, Patanjali, SankhavaramR, Surolia and Avadesha. A chitooligosaccharide specific lectin from Luffa acutangula: Physico-chemical properties and the assignment of orientation of sugars in the lectin binding site. Journal of Biosciences 1985; 8(1-2): 403-411.



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