Evaluation of Hypoglycemic Activity of Saccharum spontaneum.

M. PHARM DISSERTATION PROTOCOL

SUBMITTED TO THE

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BANGALORE

BY

THUMMAPALA HARI SHANKAR .B Pharm

UNDER THE GUIDANCE OF

DR. SHIVAKUMAR SWAMY M.Pharm., Ph.D.

H.O.D & PRINCIPAL

MALLIGE COLLEGE OF PHARMACY, BANGALORE

MALLIGE COLLEGE OF PHARMACY

#71 SILVEPURA, BANGALORE 90

Rajiv Gandhi University of Health and Sciences,

Karnataka, Bangalore.

Annexure – II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

01 / Name and Address of the Candidate / T.HARI SHANKAR
S/O Mr. T.CHINABBAI,
BHENAM QUARTERS,HATBAZAR,JATNI,
DT:KHURDA,ODISHA.
PIN:752050.
02 / Name of the Institution / Mallige College Of Pharmacy
#71 Silvepura,
Post : Chikkabanavara
Bangalore 90
03 / Course of the Study Branch / M.Pharm (Pharmacology)
04 / Date of Admission to course / 14/12/2011.
05 / Title of the Topic / Evaluation of Hypoglycemic Activity of Saccharum spontaneum.
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 / Materials 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 form your institution
in case of 7.3. / Yes
08 / List of References / Enclosure – VII
09 / Signature of the Candidate / (T.HARI SHANKAR)
10 / Remarks of the Guide / The present research work is original and not published in any of the journals with best of my knowledge upon extensive literature review. This work will be carried out in the Pharmacology laboratory by Mr. Hari Shankar under my supervision.
11 / Name and Designation of
(in Block Letters)
11.1. Guide
11.2.Signature
11.3.Co-Guide (if any)
11.4.Signature
11.5. Head of the Department
11.6.Signature / Dr. SHIVAKUMAR SWAMY
M. Pharm., Ph. D.
Principal & HOD
Mallige College Of Pharmacy,
Bangalore, Karnataka.
Dr. SHIVAKUMAR SWAMY
M. Pharm., Ph. D.
Principal & HOD
Mallige College Of Pharmacy,
Bangalore, Karnataka
12 / Remarks of the Principal
12.1. Signature / The present study is permitted to perform in the Pharmacology laboratory of our institution and the study protocol has been approved by IAEC.
(Dr. Shivakumar Swamy)

ENCLOSURE I

Brief resume of the intended work

Introduction:

The term diabetes mellitus describes a metabolic disorder. Diabetes is a chronic disease that occurs either when the pancreas does not produce enough insulin or when the body cannot effectively use the insulin it produces. Insulin is a hormone that regulates blood sugar. Hyperglycemia or raised blood sugar is a common effect of uncontrolled diabetes and over time leads to serious damage to many of the body's systems, especially the nerves and blood vessel. The effects of diabetes mellitus include long-term damage, dysfunction and failure of various organs.1

Diabetes mellitus may present with characteristic symptoms such as : Thirst, polyuria, blurring of vision, and weight loss. In its most severe forms, ketoacidosis or a non-ketotic hyperosmolar state may develop and lead to stupor, coma and, in absence of effective treatment, death. Often symptoms are not severe, or may be absent, and consequently hyperglycemia sufficient to cause pathological and functional changes may be present for a long time before the diagnosis is made.2,3.

The long-term effects of diabetes mellitus include: progressive development of the specific complications of retinopathy with potential blindness, nephropathy that may lead to renal failure, and or neuropathy with risk of foot ulcers, amputation, Charcot joints, and features of autonomic dysfunction, including sexual dysfunction.

People with diabetes are at increased risk: of cardiovascular, peripheral vascular and cerebrovascular disease. Several pathogenetic processes are involved in the development of diabetes: These include processes which destroy the beta cells of the pancreas with consequent insulin deficiency, and others that result in resistance to insulin action. The abnormalities of carbohydrate, fat and protein metabolism are due to deficient action of insulin on target tissues resulting from insensitivity or lack of insulin.3.

Reports from the International Diabetes Federation (IDF) indicate that the prevalence of diabetes mellitus has reached epidemic levels globally. Estimates for 2010 indicate that 285 million adults have diabetes in the seven regions of the IDF. These numbers represent an increase of 39 million from 2007 and an expected continued increase to 439 million in 2030. It is believed that by 2025, more than 75% of the world population with diabetes will reside in developing countries and the countries with the largest populations of adults with diabetes will include: India, China and the United States. India has 50.8 millions of people with diabetes in 2010 and expected to increase by 87 million by 2030. The highest number of deaths attributable to diabetes is expected to occur in countries with large populations-1,008,000 deaths in India4.

Report from Indian council of medical research (ICMR-INDIAB).

62.4 million Lives with diabetes in India, 77.2 million people with pre-diabetes

New figures for diabetes prevalence in India indicate that the epidemic is progressing rapidly across the nation, reaching a total of 62.4 million persons with diabetes in 2011.

Phase one results of the Indian Council of Medical Research – India Diabetes (ICMR-INDIAB) Study have provided data from three States and one Union Territory, representing nearly 18.1 per cent of the nation's population.

When extrapolated from these four units, the conclusion is 62.4 million people live with diabetes in India, and 77.2 million people are on the threshold, with pre-diabetes.5.

The present pharmaceutical drugs are either too expensive or have undesirable side effects. Treatment with sulphonylureas and biguanides are also associated with side effects4. Compared with synthetic drugs, drugs derived from the plants are considered to be less toxic with fewer side effects. However, for a number of reasons, complementary medicine has grown in popularity in recent years.

Dietary measures and traditional plant therapies as prescribed by Ayurvedic and other indigenous systems of medicine are used commonly in India. Many indigenous Indian medicinal plants have been found to be useful to successfully manage diabetes and some of them have been tested and their active ingredients isolated. The World Health Organization (WHO) has also recommended the evaluation of the plants effectiveness and conditions where we lack safe modern drugs6.

NEED FOR STUDY

On careful observation of the chemical constituents of Saccharum Spontaneum, it is observed that itcontains many chemical constituents which are already proved for their anti-diabetic activity. The extensive literature survey of Saccharum Spontaneum reveals that its anti hypoglycemic property is not scientifically tested and established.

In the present scenario the health providers looks at natural sources to maintain health conditions rather than synthetic drugs. It is also established beyond doubt that, the drugs derived from plants sources have less toxic and hence proved beneficial for ailments which require lifelong treatment.

In the recent past years many medicinal plants are screened for their hypoglycemic property and quite a few of them are already successful in entering the market.

Considering all the above points we felt, it is worth to investigate the hypoglycemic property of Saccharum Spontaneum.

ENCLOSURE II

REVIEW OF LITERATURE:-.

ScientificName:SaccharumspontaneumL
Family: Poacea.

Saccharum Spontaneum, also known variously as thewild sugar cane, false sugar cane, wild cane.

Hindi Name: Kans, Kansi, Kas. Kans grass (Saccharum spontaneum) It is a perennial grass, growing up to three meters in height, with spreading rhizomatous roots.

Common name:

Kans grass • Hindi: Kaans • Manipuri: ঈ Ee • Tamil: Pekkarimpu • Telugu: Kaki ceruku • Marathi: Kamis • Bengali: Kansh • Kannada: Kadu kabbu • Malayalam: Nannana • Gujarati: Kansado Oriya: କାଶତଣ୍ଡି kāśataṇḍi), In the Terai-Duar savanna and grasslands, a lowland ecoregion at the base of the base of the Himalaya range in Nepal, India, Bangladesh and Bhutan.

General Description: It grows as wasteland weed. It is considered as valuable medicinal herb in traditional systems of medicine in India. It is popular folk medicine.

Distribution.7.
Gregarious, in open areas, in low and medium altitudes, throughout the Philippines.
Occurs from India to southern China and through Malaya to Polynesia
It is a coarse, erect, perennial grass, with stout underground rootstock growing to a height of 1 to 3.5 meters. Leaves are harsh and linear, 0.5 to 1 meter long; 6 to 15 mm wide. Pannicles are white and erect, measuring 15-30 cm long, with slender and whorled branches, the joints covered with soft white hair. Spikelet’s are about 3.5 mm long, much shorter than the copious, long, white hairs at the base.

Constituents.9,10.
Phytochemical screening yielded: quinones, terpenes, alkaloids, flavonoids, saponins, tannins, carbohydrates, protein, coumarin, phenol, steroids and glycosides.

Studies.7.

Mohammad Khalid et al., conducted a study on Pharmacognostical Evaluation and Qualitative Analysis of Saccharum spontaneum (L.) Root.They studied various parameters like macroscopy; Microscopy, fluorescence analysis as well as extractive value and quantitative phytochemical screening of different extractives were studied. The major components of the extractives like total phenolic, total flavonoids were also estimated respectively. The characteristic of microscopy, physicochemical, fluorescence analysis and quantitative chemical screening were performed in root extractives of the plant material as a mean of authentication.8.

Suresh kumar et al., conducted a study on Pharmacognostic and Preliminary Phytochemical Investigations on the stem of Saccharum spontaneum. Scientific information on their pharmacognosy, Phytochemistry and pharmacology are very scant. The study describes some pharmacognostical and preliminary phytochemical investigations undertaken on the stem of one of those species namely Saccharum spontaneum. The samples for research were collected from Vellore, Tamil Nadu, India and authentificated by Dr.P.Jayaraman Ph.D., a director of plant Anatomy Research Centre, and then subjected for morphological, microscopical and physicochemical analysis. They found that the Screening yielded the presence of quinines, alkaloids, tannins, carbohydrates, protein, coumarin, phenol, steroid and glycosides.9.

Suresh kumar et al., conducted a study on Psychopharmacological activites of various extracts of the stem of Saccharum spontaneum by taking rats and found that reduction in the motor activity indicates C.N.S.depressant property of the drug. The locomotor activity of normal rat animal shows the C.N.S depressant activity (6.53%). The ethanol extract shows the 10 times (62.0%) more activity than the control. Whereas other extract like Aqueous extract (21.9%) shows the 4 times more activity and chloroform extract shows the 2 times (10.0%) more C.N.S. depressant activity than the control. The Ethanol and Aqueous extract of Saccharum spontaneum must have a significant C.N.S. depressant activity than chloroform and aqueous extracts when compared to control. The above extracts show the antishycotic activity but are less when compared with standard drug of chlorpromazine. It derives that alkaloids, tannins, steroids and glycosides are present in the extract which may possibly responsible for the psychopharmacological action.10.

Farhana Alam Ripa et al., conducted a study on In Vitro Antimicrobial, Cytotoxic and Antioxidant Activity of Flower Extract of Saccharum Spontaneum Linn . Chloroform extract showed antioxidant activity with IC50 value of 51.04 ug/ml (vs ascorbic acid 41.04). Crude extract showed cytotoxic activity using Brine shrimp lethality assay with LC50 of 6.63 ug.mL (vincristin 10.64). Antimicrobial activity was exhibited against gram-positive and gram-negative pathogenic bacteria and against three tested fungi.11.

Mohammad Khalid,conducted a study on Free Radical Scavengering And Total Phenolic Content Of Saccharum Spontaneum L. Root Extract, The methanolic extract were prepared and screened for in-vitro antioxidant activities using 1, 1-Diphenyl-2-picrylhydrazyl (DPPH) scavenging activity method. Also, extract were assessed for thiocyanate, reduction potential and nitric oxide scavenging activity. They found that the total phenolic content was 351.25 ± 1.31 μg mL-1 however the flavonoid content was 48.60 ± 2.17 μg mL-1.12.


Tyagi conducted a study on Pulp and Paper Making Characteristics of Saccharum spontaneum.Study showed S. spontaneum is a bulky material with lower extractives and lignin content and higher holocellulose content with a good response towards multistage bleaching sequence and a pulp of high brightness ceiling.13.

Bi Umbrin Ilyas et al., conducted a study on regional agrowastes such as Vigna mungo, Saccharum spontaneum and Brassica campestris were collected and biohydrolysis of these substrates for cellulase production were carried out by Aspergillus niger. Proximate composition of each agrowastes was analyzed based on dry weight, to have an insight view of their chemical composition. Cellulose content of S. spontaneum and B. campestris was calculated as 43.1 ± 2.8% and 39.4 ± 3.1%, respectively, while cellulase activity of A. niger was found to be higher on S. spontaneum. Low lignin to cellulose ratio of S. spontaneum made it a preferred substrate for cellulase production. However, high lignin content of B. campestris made the cellulose inaccessible and resulted in poor yield of enzyme. They found that S. spontaneum has a great potential to serve as a cheaper, easily available and reasonable substrate for cellulase production. 14.

ENCLOSURE III

OBJECTIVE OF THE STUDY:

The present research work is an attempt to establish the possible hypoglycemic efficacy using aquous and ethanol plant extract of Saccharum Spontaneum in rats with the following objectives.

1.  To collect and authentication of the plant.

2. To prepare aqueous and 70% ethanol extract of Saccharum Spontalanum.

3. Test for phytoconstituents of the extract.

4. Screening for hypoglycemic activity on normal & diabetic albino rat.

ENCLOSURE IV

MATERIALS:

7.1 Source of data

The work is aimed to generate data from the experiments to be conducted at Pharmacology laboratory of our institution.

The experiement, which involves the following steps:

Plant Extract

·  Collection and authentication of plant of Saccharum spontaneum.

·  Extraction of Saccharum spontaneum.

·  Qualitative estimation of phytochemical constituents.

·  Screening for hypoglycemic activity.

Animals

·  Adult Albino rats will be used for this purpose, and they will be obtained from animal house of Mallige College of Pharmacy.

·  Animal clearance will be obtained from institutional animal ethical committee for experimental purpose.

·  They will be maintained under laboratory condition with controlled environment of temperature, humidity being provided with standard diet and water ad libitum.