RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES, BANGLORE, KARNATAKA

ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION
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NAME OF THE CANDIDATE

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MAMATHA M.K

DEPT. OF PHARMACOGNOSY & PHYTOCHEMISTRY

K.L.E.S’s COLLEGE OF PHARMACY, BELGAUM-10
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NAME OF THE INSTITUTION

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K.L.E. SOCIETY’S COLLEGE OF PHARMACY, JNMC CAMPUS,

NEHRU NAGAR,

BELGAUM-10

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COURSE OF THE SUBJECT STUDY

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MASTER OF PHARMACY IN

PHARMACOGNOSY & PHYTOCHEMISTRY
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DATE OF ADMISSION OF COURSE

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JUNE 2007

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TITLE OF THE TOPIC:

“PHYTOCHEMICAL INVESTIGATION AND ANTIDIABETIC ACTIVITY OF THE LEAVES OF HOLOPTELEA INTEGRIFOLIA (ROXB.) PLANCH”.
6. / BRIEF RESUME OF THE INTENDED WORK.
6.1 NEED FOR THE STUDY:
The world health organization (W.H.O) has estimated that perhaps 80% of the world population relies chiefly on traditional medicines for primary health care needs and in 1998, suggested appropriate guidelines for the use of herbal medicines for various chronic disorders 1.
The Diabetes mellitus is considered to be a serious endocrine syndrome. In many countries it is traditional to use medicinal plants to control diabetes in anti hyperglycemic effect of several plant extract and herbal formulations which are used as anti-diabetic remedies has been confirmed. The synthetic hypoglycemic agents can produce serious side effects including hematological effect, coma and disturbances of the liver and kidney. In addition they are not suitable for use during pregnancy, compared with synthetic drug, which are derived from plants are frequently considered to be less toxic with some side effects. Therefore the search for more effective and safer antihyperglycemic agents has become an area of active research 2.
Since from the ancient times a number of herbal medicine have been used in the treatment of this disease and many studies have been carried out in the search of a suitable plant drug that would be effective in diabetes mellitus. There is increasing demand by patients to use the natural products with anti-diabetic activity.3
In the Ayurvedic treatments, medicines consists of plant products, which have different pharmacological activities, which have been revealed in many literatures.4
The plant of Holoptelea integrifolia reveals that various parts such as bark, seed, leaves, latex are found to be useful in Ayurvedic system of medicine for different pharmacological activity such as inflammations, dyspepsia, flatulence, colic, helimenthiasis, vomiting, skin diseases, leprosy &diabetes.4&5
The latex and the extract of dried bark, seed and leaves of Holoptelea integrifolia has not been scientifically investigated for anti-diabetic activity. Hence the present study is under taken for the phytochemical investigation of the leaves of Holoptelea integrifolia and to evaluate its claimed anti-diabetic activity. 5
6.2 REVIEW OF LITERATURE

Holoptelea integrifolia (Roxb.) Planch. (Family – Ulmaceae) is a medium sized to large glabrous deciduous tree, 15-25 m in height with whitish or yellowish grey bark.4

The tree is distributed through out India up to an altitude of 660 m. lower ranges of Himalaya from Jammu to Oudh.6

It is a medium sized tree with white colored branches. Holoptelea integrifolia thrives in deep porous soil with good drainage but becomes stunted and crooked on poor shallow soil.7

The plant is known by various names in different languages as follows 6

Sanskrit : Chirabilva
English : Indian elm, Kanju
Hindi : Cibil, Kanju
Kannada : Tapasigida, Thavasi
Telugu : Tapasi, Thapasi
Tamil : Avali, Kanji
Malayalam : Avil, Aval
It is sometimes grown on the road side. Bark grey, pustular, exfoliating in some what corky scales; Leaves elliptic-ovate, acuminate, base rounded or subcordate, flowers greenish yellow, polygamous, in short racemes or fascicles; Fruit a sub-oricular samara with membranous wing, edible; seed flat. The bark when cut and leaves and twigs when crushed emit an unpleasant odour.8
It is a medium sized tree with white coloured branches. Leaves 2.5 to 20cms long and 5 to 10 cm broad, egg shaped, pointed with transparent spots. Dried leaves exhibit small elevations on the ventral surface. Flowering season is summer followed by fruiting. Leaves and trunk have an offensive odour.5
Chemical constituents:6
Stem bark:
Two terpenoid fatty acid esters holoptelin A and B, 2- aminonaphtaqinone, friedelin, epifriedelinol, -sitosterol and its -D-glucose.
Heartwood:
-sitosterol, 2,3-dihydroxyolean-12-en-28 oic acid and hederagenin.
Leaves:
Hexacosanol, octacosanol, -sitosterol and -amyrin.
Dried seeds:
Carbohydrates, pigments, oils, acids, glycosides, sterols, tannins, proteins, free amino acids, major fatty acids-palmitic acid, oleic acids, myristic, stearic, linoleic and linoleic acids and steroids--sitosterol and stigmasterol.
Pollens:
Histamine and 5-hydroxytryptamine.
Roots:
24- ethyl-cholest-22-en-3alpha-ol etc.9
Seed cake: the seed cake contains N, 10.3 lysine, 3.3.glutamicacd, 13.0. and histidine 1.3%. Triterpenoid fatty acid esters, friedelin.,etc.10
The bark and leaves are bitter, astringent, acrid, thermogenic, anti inflammatory, digestive, carminative, laxative, anthelmintic, repulsive and dyspepsia seeds are useful in infected ulcers and as deodorant for foul smell of body.6
Review of literature reveals that Holoptelea integrifolia and Iris versicolor have been reported antiobesity properties.11
Ricinus communis and Holoptelea integrifolia are important pollen allergens of India.12
Biochemical analysis in the form quantitative estimation of carbohydrates, proteins, nucleic acids, minerals, ash and moisture of the pollen of two allergenic plants, i.e. carica papaya and Holoptelea integrifolia has been described.13
The stem bark of khakro (Butea monosperma), Ingoriyo (Balanites aegyptica) and HSoloptelea integrifolia is crushed and dropped in water for use as fish poison.14
B.lingulata and H.integrifolia have been reported screening of antiviral activity.15
H.integrifolia used as control white fly bemisia tabaci gennadius 73 in lal pani nursery dehra dun.16
To study cross reactivity among pollen of ulmaceae from geographically distant locations.An attempt to study cross reactivity with Elisa inhibition in these patients further indicated pan allergenicity. It is concluded that cross reactivity among pollen of different geographical locations poorly observed.17
6.3 OBJECTIVE OF THE STUDY:
The objectives of the present study are:
  • The extensive review of literature on Holoptelea integrifolia.
  • Collection of the plant / preparation of herbarium sheet of the plant Holoptelea integrifolia.
  • Extraction of the leaves of Holoptelea integrifolia using polar and non polar solvent.
  • Detailed phytochemical investigation / tests of the plant extracts to be carried out.
  • Identification / separation of major chemical constituents using TLC, column chromatography and HPTLC techniques from various extracts.
  • Antidiabetic activity of the various extracts of Holoptelea integrifolia and antioxidant activity shall be screened.
  • Formulation of various extracts showing significant activity.

7. / MATERIALS AND METHODS:
7.1 SOURCE OF DATA:
Sources of data are from the laboratory experiments, Holoptelea integrifolia which involve successive solvent extraction of plant, with their phytochemical investigation and anti-diabetic activity on experimental animals such as Albino Rats.
7.2 METHOD OF COLLECTION OF DATA: (including sampling procedure if any)
A. Collection of Drug:
The leaves of Holoptelea integrifolia. are collected from local areas of Belgaum, Dharwad district.
B. Standardization of plant parts:18
1. Botanical Evaluation:
Organoleptic evaluation: colour/ odour/ taste/ size/ shape/ texture/ foreign matter and visual microscopy etc.
2. Physicochemical Evaluation:
Ash values (acid insoluble ash, water insoluble ash, sulphated ash), extractive values (Alcohol soluble, and water soluble), loss on drying, moisture content etc.
3. Phytochemical evaluation:
Successive Solvent Extraction:
A leaves of Holoptelea integrifolia, plant will be powdered to required particle size. The powdered material will be subjected to successive solvent extraction with polar and nonpolar solvents using soxhlet assembly, each time before extracting with next solvent the powder material will be dried in hot air oven below 50C. Each extract of the drug will be concentrated by distillating off the solvent and then evaporated to dryness on hot water bath. The extract obtained with each solvent will be weighed, its percentage will be calculated in terms of air dried weight of the plant material. The colour and consistency of each extract will be noted.
- The each extract will be subjected to phytochemical investigation for the identification of phytoconstituents.19
- Separation and isolation of phytoconstituents by chromatographic techniques.
- Subject the isolated compound to specific studies.
C. Anti-diabetic activity:
Pharmacological Studies:
  1. Determination of LD50 (for test compound): The method of OECD Guidelines will be adapted for the determination of LD5020 Albino mice of either sex weighing between 20-25g will be used. The extract will be administered by appropriate route.
  2. Anti-diabetic activity: The anti-diabetic activity will be assessed by alloxan induced diabetes in Albino Rats of either sex.
The study will be carried out by the test extract and Standard drug. In which the animals will be segregated into eight groups of either wister / SD Albino rats each, taking into concentration of the diabetic blood sugar level, one group is normal control and other is diabetic control along with polar and non polar solvent extracts which will be compared with Standard group.
The blood glucose level will be measured in all the group using Glucometer21 of Anti-diabetic activity will be statistically analysed with the help of various biostatical methods.
7.3 DOES THE STUDY REQUIRE ANY INVESTIGATION OR INTERVENTION TO BE CONDUCTED ON PATIENTS OR OTHER HUMAN OR ANIMALS? IF SO, PLEASE MENTION BRIEFLY.
Yes. The above study requires investigation on rats. The experiments will be conducted on those animals in anaesthetized condition. They are sacrificed at the end of the study using proper anaesthetic agent.
7.4 HAS THE ETHICAL CLEARANCE BEEN OBTAINED FROM YOUR INSTITUTION IN CASE OF 7.3?
The study has been cleared from ethical committee of the institution.
(certificate enclosed)

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8. / REFERENCES:
  1. Sudha S, Bindu R, Joyce G, Amit A, Venkataraman BV. Pharmacological Interaction of Centella asiatica and Bacopa monnieri with anti-epileptic drugs-an experimental study in rats. Journal of natural Remedies 2005; 5 (1) : 63-9.
  2. Pari L, Ramakrishna R. Anti hyperglycemic effect of Diamed, a herbal foundation in experimental diabetes in rats. Journal of Pharmacy and Pharmacology 2001; (53): 1139-43.
  3. Jude E, Okokon bassey S, Antia. Hypoglycemic and anti diabetic activity of setaria megaphylla on normal and alloxan induced diabetic rats. Journal of Natural remedies 2007; 7(2): 220-24.
  4. Arya Vaidyasala. Indian medicinal plants. Chennai: Orient Longman Limited;1995; (2): 162-64.
  5. Vaidya V.M.Gogte. Ayurvedic pharmacology & therapeutic uses of medicinal plants. Swami Prakashananda Ayurveda research centre (SPARC). Chaukhambha publications; 2000: 607-08.
  6. Sharma PC, Yelne MB, Dennis TJ. Database on medicinal plants used in ayurveda, Central council for research in ayurveda & sidha, New Delhi; 2001; (2):171-76.
  7. Wealth of India. A Dictionary of Indian raw materials and Industrial products. New Delhi ; publication and information,CSIR;1959;(5): 109-10.
  8. Kirtikar KR & Basu BD. Indian medicinal plants. 2nded. Dehradun: International book distributors; 1975;( 3): 292-94.
  9. Jain R, Alam S. 24-ethyl-cholest-22-en-3alpha-ol and other constituents from the roots of Holoptelea integrifolia. Indian journal of chemistry 1991; 37(4):192-91.
  10. Irfanalikhan, Atiyakhanum. Ethnomedicine and Human welfare. Ukaaz publications 2005;(4):154.
  11. Bambole VD. Effect of some medicinal plant preparations of adipose tissue metabolism. Ancient science of life 1988 ; 8(2): 117-24.
  12. Singh AB, Malik P, Gangal SV, et al. Unique and shared allergenic components in pollen of two unrelated taxa. Indian Journal of Aerobiology 1991; 4(1&2): 30-31.
  13. Datta BK, Mandal S, Bhattacharya GN. Biochemical analysis of the allergenic pollen of Carica papaya L. and Holoptelea integrifolia (Roxb.) Planch. Indian journal of aerobiology 1992; (special volume):113-15.
  1. Punjani BL. Fishing and hunting devices used by tribe kathodi of vijayanagar. Ethnobotany 2002; 14(1&2): 129-30.
  2. Rajbhandar M, Wegner U, Julich M, Mentel R. Screening of Nepalese medicinal plants for antiviral activity. Journal of Ethnopharmacology 2001; 74(3): 251-55.
  3. Misra RM, Mishra RM, Rawat DS. Control of white fly bemisia tabaci gennadius 73 in Lal Pani nursery. Dehradun: Annals of forestry 2000; 8(1): 159-60.
  4. Singh AB, Panzani R, Shah A, Sharma S, Kumar P. Assessing cross reactivity among pollen of ulmaceae from geographical distinct locations.Inian Journal of Aerobiology 2001; 14(1&2): 44.
  5. Khadelwal KR. Practical Pharmacognosy Techniques and Experiment. 13th ed., Pune: Niraliprakashan; 2005: 157-60.
  6. Kokate CK. Practical Pharmacognosy. 4th ed., Delhi: vallabh prakashan; 1994: 107-11.
  7. OECD/OCDE, OECD guidelines for the testing of chemicals revised draft guidelines. Acute oral toxicity-Acute toxic Class Methods, Revised document, October 2000:2423.
  8. Kandasamy Murugesh,Veerendra Yeligar, Deepak kumar dash, Panaki sengupta, Bhim chadra maiti, et al.Antidiabetic,Antioxidant & Antihyperlipidemic status of Heliotropium zeylanicum extract on Streptozotocin-Induced diabetes in rats. Biol, Pharm, Bull. Pharmaceutical society of Japan 2006; 29(11):2202-205.

9. / SIGNATURE OF CANDIDATE:
10. / REMARKS OF THE GUIDE: The above information and literature has been extensively investigated, verified and was found to be correct. The present study will be carried out under my supervision and guidance.
11. /
NAME & DESTINATION OF
(in block letters)
11.1GUIDE:
11.2 SIGNATURE:
11.3 CO-GUIDE:
11.4 SIGNATURE:
11.5 HEAD OF THE DEPARTMENT:
11.6 SIGNATURE / Mr. PRAMOD H.J M.pharm.
Assistant professor,
Dept. of Pharmacognosy & phytochemistry,
K.L.E.S’s College of Pharmacy,
Belgaum-10.
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Dr. SUNIL S. JALALPURE M.Pharm, Ph.D.
Associateprofessor & head,
Department of Pharmacognosy & phytochemistry
K.L.E.S’s College of Pharmacy,
Belgaum-10.
12. /
12.1 REMARKS OF THE CHAIRMAN & PRINCIPAL:
12.2 SIGNATURE: / The above mentioned information is correct and I recommend the same for approval.
Dr. F V. Manvi. M. Pharma, Ph D.
Principal,
K.L.E.S’s College of Pharmacy,
Belgaum-10.

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