RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BANGLORE

Annexure-II

PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION

01 / Name and Address of the Candidate / Thakor Pushpendrasinh J.
S/O Jayendrasinh
In - vanta
At-post : kheda
pincode: 387411 Dist - kheda
State –Gujarat
02 / Name of the Institution / T.M.A.E. Society’s
S.C.S.College of Pharmacy,
Harapanahalli-583131
(Karnataka)
03 / Course of the Study Branch / M. Pharm (Pharmacology)
04 / Date of Admission to course / 14-05-2007
05 / Title of the Topic / COMPARATIVE ANTIDIABETIC STUDY OF MORINGA OLEIFERA PODS AND LEAVES
06 / Brief resume of the intended work
6.1 / Need for the Study / Enclosure – I
6.2 / Review of the Literature / Enclosure – II
6.3 / Objective of the study / Enclosure – III
07 / Material and Methods
7.1 / Source of Data / Enclosure – IV
7.2 / Methods of collection of data / Enclosure – V
7.3 / Does the study require any investigations on animals?
If yes give details / Enclosure –VI
7.4 / Has ethical clearance been obtained from your institution in case of 7.3 / Yes, Registration No: 157/1999/CPCSEA
(Copy enclosed)
8 / List of reference / Enclosure –VII

ENCLOSURE-I

6. Brief resume of intended work:

6.1 Need For the Study:

Diabetes mellitus is one of the common metabolic disorder and 1.3% of the population suffers from this disease throughout the world1. Insulin and oral hypoglycemic agents like sulphonylureas and biguanides are still the major players in the management of the disease. However, complete cure of the disease has been eluding physicians for centuries and the quest for the development of more effective antidiabetic agents is pursued relentlessly.

Many herbal products, including several metals and minerals have been described for the cure of diabetes mellitus in ancient literature. Herbal preparations alone orin combination with oral hypoglycemic agents sometime produce a good therapeutic response in some resistant cases where modern medicines alone fail2. Currently available treatment is far from satisfactory and is expensive.In spite of the progress in treatment of diabetes mellitus by introduction of various synthetic drugs, search for novel natural drugs continues because of several therapeutic complications associated with existing therapy. Herbal medicines are particularly used by the traditional practitioners for the treatment of diabetes since the ancient time but they remain to be scientifically investigated.

Moringa oleiferalam.is belongs to family moringaceae, commonly known as “Drumstick.” Almost every part of plant is of value for food. Different parts of this tree have been used in folklore medicine for the various ailments.Leaves and pods are particularly recommended in the folk medicine for the diabetes3. The plant is said to be having hepatoprotective4, antioxidant5 ,wound healing and antipyretic6, hypocholesteric7, antimicrobial activity8,9. The stem bark of Moringa oleifera has been also reported to possess antihyperglycemic activity10. However, no scientific dataon antidiabetic activity of leaves and pods of the Moringa oleifera has been reported so far. Therefore, thepresent study is undertaken.

ENCLOSURE-II

6.2 Review of Literature:

The plant Moringa oleifera(family: Moringaceae),commonly known as drumstick tree or horseradish tree3, it is known as Drumstick in English, Saragvo in Gujrati, Soanjna in Hindi and Nugge in Kannada.

Description: It is perennial softwood tree with timber of low quality; rather slender with drooping branches; branches and stems brittle, with corky bark; leaves feathery, pale green, compound, tripinnate, 30-60 cm long, with many small leaflets, 1.3-2 cm long, 0.6-0.3 cm wide, lateral ones somewhat elliptic, terminal one obovate and slightly larger than the lateral ones; flower fragrant, white or creamy-white, 2.5 cm in diameter, borne in sprays, with 5 at top of the flower; with yellow steam and brown, triangular pods11.

In India, the plant is propagated by planting limb cutting 1-2 cm long, from June to August, preferably. The plant starts bearing pods 6-8 months after planting but regular bearing commenced after second year. Fruiting may peak twice in year first between March and April and again in September and October. Seed gathered in March and April11.

Chemical Constituents:

Leaves of Moringa oleifera contains two nitrile Glycosides, niazirin, niaziminin12, glucosinolates, flavonoids, anthocyanin, and proanthocyanidin, cinnamatesβ-sitosterol13.

Pods reported to contain nitriles, an isothiocynate and thicarbamate12, β-sitosterol14 etc.

Rootsreported to be having mainly two alkaloids: moringine and moringinine11.

Medicinal uses3:

Pods are used in hypertension, diabetes, and joint pain.

Leaves are used in diabetes, ulcer, wounds, diarrhoea etc.

Rootsare used in cardiovascular disorders,ulcer.

Seeds are used in worm infestation, cancer, and ulcer etc.

.

ENCLOSURE-III

6.3 Objectives Of the study:

  1. This study is planned to investigate and validate the anti-diabetic activityagainst alloxan diabetic model in rats.
  2. In view of this, the leavesand pods of Moringa oleifera have been selectedfor the present investigation.

ENCLOSURE-IV

7. Material & methods:

7.1 Source of data:

Whole experimental work is designed to generate data from the laboratory i.e onexperimental animals. Albino rats and mice will be used for this purpose.

The experiment which involves.

1)Extraction of pods and leaves of Moringa oleifera by Soxhlet’s apparatus using 95% ethanol.

2)The preliminary phytochemical constituents present in the leaf and pod extracts will be carried out by following literature reported chemical tests.

3)The acute toxicity of extracts will be studied on micefollowing fixed dosemethod of CPCSEA.

4)Crude extract of Moringa oleifera leaves and pods for anti-diabetic activitywill be screened against alloxan induced diabetic rats.

ENCLOSURE-V

7.2 Method of collection of data:(Including sampling procedure)

a)Phytochemical Studies:

The leaves and pods ofMoringa oleifera will be collected from the surrounding gardens of Harapanahalli.They wiil be cleaned, shade dried,coarse powdered, and subjected toextraction with 95%ethanol by using soxhlet’s apparatus. There after the extract will be concentrated using rotaryflash evaporator. The crude extract thus obtained will be analysed for the presence of phytochemical constituents (Preliminary phytochemical screening)15.

b) Pharmacological Studies:

1. Acute toxicity study of crude extract16.

For this purpose female albino mice (25-30 g) will be used. Fixed dose method (OECD guideline no. 420) of CPCSEA will be adopted.

2. Dosedependant (1/25th, 1/10th, 1/5thof LD50 cutoff value)comparative evaluation of leaf and pod extracts of Moringa oleifera will be carried out on alloxan induced diabetic rats.

Antidiabetic activity includes17:

(A)Estimation of the following biochemical parameters:

a) Glucose

b) Urea

c) Creatinine

d) Cholesterol

e) Protein

(B) Morphological studies:

-Recording body weight of animal.

(C)Histopathological studies of pancreas.

Statistical analysis

The results obtained from the above investigation will be subjected to statistical analysis using one way ANOVA followed by Tukey- Kramer Multiple Comparision test.

Total duration of the completion of the project work would be of ten months.

1. Collection of the plant material.1 month

2. Duration of experimentation including

Extraction process5 months

3. Literature survey2 months

4. Thesis writing including communication

of the research paper for the publication 2 months

ENCLOSURE-VI

7.3Dose the study requires any investigation or investigations to conduct on patients or other humans or animals? If so please describe briefly.

Albino rats (150-200g, Wistar strain) of either sex will be used for the evaluation of anti-diabetic activity. Where as albino mice (25-30g, Swiss strain) will be utilized for the study of the acute toxicity.

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

The present study protocol is approved from Institutional Animal Ethics Committee (IAEC certificate enclosed).

ENCLOSURE-VII

8.0 List of References:

  1. Raghunathan M, Raghunathan N. Diabetes mellitus and vitamin D. Nutrition News 1992; 13: 4p.
  1. AnturlikarSD, Gopumadhavan S, Chauhan BL, Mitra SK. Effect of D-400, A Herbal formulation on blood sugar of normal and alloxan induced diabetic rats. Indian Journal Physiol Pharmacol. 1995; 39: 95-100.
  1. Fahey JW. Moringa oleifera: A review of the medical evidence for its nutritional, therapeutics, and prophaylactic properties. PartI. Trees for Life Journal. 2005;1(5):1-15.
  1. Nadro MS, Arungbemi RM, Dahiru D. Evaluation of Moringa oleifera leaf extract on alcohol-induced hepatotoxicity. Tropical Journal of Pharmaceutical Research. 2006; 5(1): 539-44.
  1. Siddhuraju P, Becker K. Antioxidant properties of various solvents extracts of total phenolic constituents from three diffrent agroclimatic origins of Drum stick tree (Moringa oleifera lam.) leaves. J Agric Food Chem. 2003; 51(8): 2144-55.
  1. Hukkeri VI, Nagathan CV, Karadi RV, Patil BS. Antipyretic and wound healing activities of Moringa oleifera lam. in rats. Indian Journal of Pharmaceutical Sciences. 2006; 68(1): 124-26.
  1. Ghasi S, Nwobobo E, Ofili JO. Hypocholesterolemic effects of crude extract of leaf of Moringa oleifera Lam. in high-fat diet fed Wistar rats. Journal of Ethanopharmacology. 2000; 69: 21-25.
  1. Doughari JH, Pukuma MS, De N. Antibacterial effects of Balanites aegyptiaca L. and Moringa oleifera lam. on salmonella typhi. African Journal of Biotechnology. 2007; 6 (19): 2212-15.
  1. Umar RM. Antimicrobial activity of small Moringa oleifera leaves. Journal of Islamic Academy of Sciences. 1998; 11(1): 27-32.
  1. Kar A. Choudhary BK, Bandyopadhyay NG. Comparative evaluation of hypoglycaemic activity of some Indian medicinal plants in alloxan diabetic rats. Journal of Ethanopharmacology. 2003; 84(1):105-08.
  1. Kirtikar KR, Basu BD, Indian Medicinal Plants.2nd ed. New cannaught Place Dehra Dun. Bishen Singh, Mahendra Pal singh, 1975; 1:676p.
  1. Faiz S, Siddiqui BS, saliim R, Siddiqui S, Aftab K. Isolation and structure elucidation of new nitrile and mustard oil glycosides from Moringa oleiferaand their effect on blood pressure. Journal of Natural Products. 1994; 57(9): 1256-61.
  1. Bhoomika RG, Babita B Agarwal, Ramesh K Goyal, Anita A Mehta. Phyto-Pharmacology of Moringa oleifera - An overview. Natural Product Radiance; 2007; 6(4): 347-53.
  1. Faiz S, Siddiqui BS, saliim R, Siddiqui S, Aftab K Shaheen F, Gilani A . Hypotensive constituents from the pods of Moringa oleifera. Planta Med, 1998; 64: 225-28.
  1. Dr. Kandelwal KR. Practical pharmacognosy. 11th ed. Nirali prakashan, Pune; 2004: 149-56.
  1. Mrs Prema Veeraragavan. Expert consultant, CPCSEA, OECD guideline no.420, 2000.
  1. Bhavapriya v. et.Al. Biochemical studies on hypoglycemic effect of Aavirai kudineer. An herbal formulation in alloxan induced diabetic rats. Indian Journal of Experimental Biology, 2001; 39: 925-28.