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Title:
A COMPOSITION WITH ANTI HYPERGLYCEMIC AND ANTIOXIDANT ACTIVITY OBTAINED FROM THE EXTRACT OF PARTS OF COCONUT AND A METHOD OF PRODUCING THE SAME
Document Type and Number:
WIPO Patent Application WO/2013/179308
Kind Code:
A2
Abstract:
The invention relates to a composition with anti hyperglycemic, antidiabetic and antioxidant activity obtained from the extract of parts of coconut tree selected from roots, bark, trunk, leaves and coconut shell and husk or combinations thereof and a method of producing the same. The composition is polyphenols in the range of 5-80%; flavonoids in the range of 0.2 - 10%; tannins in the range of 10- 60%; saponins in the range of 1 - 30%; alkaloids in the range of 1 -25%; anthocyanins in the range of 0.1 -5%, proteins in the range of 1 -10% and carbohydrate in the range of 1 -30%. A method of producing said composition mainly comprising of extracting with solvents selected from water, hexane, methanol, ethanol, isopropanol, n-butanol, methyl acetate, ethyl acetate, propyl acetate, n- butyl acetate and combinations thereof to obtain the said extract followed by concentrating and drying to obtain in dry powder form. It is administered to patients in the form of oral dosage and in the form of infusion, injection and ampoule.

Inventors:
ANTONY BENNY (IN)
Application Number:
PCT/IN2013/000342
Publication Date:
December 05, 2013
Filing Date:
May 29, 2013
Export Citation:
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Assignee:
ANTONY BENNY (IN)
Foreign References:
JP2006083146A2006-03-30
CN102038813A2011-05-04
Other References:
ZHOU RONGHAN ET AL. PLANT CHEMOTAXONOMY. October 2005, pages 1231 - 1232
TAN, PINXIN ET AL.: 'Research Advances in Antioxidant Composition of Botanical Extracts and Their Action Mechanisms.' FOOD SCIENCE. vol. 31, no. 15, 2010, pages 288 - 292
ISHA, DEBMANDAL ET AL.: 'Coconut (Cocos nucifera L.: Arecaceae): In health promotion and disease prevention.' ASIAN PACIFIC JOURNAL OF TROPICAL MEDICINE. 20 March 2011, pages 41 - 274
Attorney, Agent or Firm:
PUTHENPARAMPIL MANI, Georgekutty (EF 7/10 Vasanth Nagar, Palarivattom,Cochin - 5, Kerala State, IN)
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Claims:
1. A composition with anti hyperglycemic, antidiabetic and antioxidant activity obtained from the extract of parts of coconut tree selected from roots, bark, trunk, leaves and coconut shell and husk or combinations thereof comprising polyphenols in the range of 5-80%, flavonoids in the range of 0.2 - 10%; tannins in the range of 10- 60%; saponins in the range of 1 - 30%; alkaloids in the range of 1 -25%, anthocyanins in the range of 0.1 -5%, proteins in the range of 1 -10% and carbohydrate in the range of 1 -30%.

2. A composition as claimed in claim 1 , wherein it can be packaged in the form capsule, tablet, granule, sachet, powder, paste, ointment, solution, suspension, emulsion, as well as in the form of infusion, injection and ampoule.

3. A method of producing the composition as claimed in claim 1 from coconut tree selected from roots, bark, trunk, leaves and coconut shell and husk or combinations thereof comprising the steps of;

a) selecting the raw material and cleaning to make it free of any foreign mater like dirt,

b) drying the cleaned material at a temperature ranging from 100 to 150 degree C,

c) grinding or crushing the dried material into powder form, *

d) extracting with solvents selected from water, hexane, methanol, ethanol, isopropanol, n-butanol, methyl acetate, ethyl acetate, propyl acetate, n-butyl acetate and combinations thereof to obtain an extract enriched with polyphenol including flavonoids, tannins and also enriched with saponins, carbohydrates, anthocyanins, proteins and alkaloids,

e) concentrating and drying the resultant extract enriched with polyphenol including flavonoids, tannins and also enriched with saponins, carbohydrates, anthocyanins, proteins and alkaloids to obtain in dry powder form.

4. A method of producing the composition as claimed in claim 3, wherein the process steps comprising:

a) cleaning, drying, and powdering the roots of coconut tree, b) extracting with water by refluxing the mixture prepared by adding water in an amount ten times the quantity of powder for about one hour to obtain a first residue and supernatant,

c) further extracting the resultant first residue two more times with ten times the quantity of water at each time and separation of the residue and supernatants, d) pooling all the supernatants and concentrating preferably in an Agitated thin film evaporator (ATFE) at a temperature of 85°C to form concentrated water extract,

e) drying the concentrated water extract under vacuum at above 500 mm of mercury to get dried powder,

5. A method of producing the composition as claimed in claim 4 wherein the powder obtained in step e) is further subjected to the purifying process steps comprising:

a) dissolving the resultant powder obtained in step e) above in water and centrifuged for about 10 minutes to form supernatant and residue.

b) further purifying the resultant supernatant preferably by subjecting to chromatographic method of purification more preferably by loading on the

HP20 column.

c) washing the column with water and collect the water part followed by washing the column with methanol and collecting the methanol part, d) evaporating each part preferably in an agitated thin film evaporator (ATFE) at about 85°C to form concentrated extract of each part,

e) concentrating of each part preferably in a vacuum stripper and dried separately under vacuum at above 500 mm of mercury to form purified dried product in powder form.

6. A method of producing the composition as claimed in claim 4 wherein the powder obtained in step e) is subjected to further purification process steps comprising:

a) dissolving the resultant powder obtained in step e) above in water, b) adding hexane in the resultant aqueous mixture, shake well and separate into two phases -aqueous and hexane and collect the hexane phase, c) adding ethyl acetate to the aqueous phase and shake well and separate the two phases -aqueous and ethyl acetate and collect the ethyl acetate phase, d) adding butanol to the aqueous phase and shake well and separate the two phases -aqueous and butanol and collect the butanol part,

e) concentrating each part separately in an agitated thin film evaporator (ATFE) to form concentrated extract of each part and followed by drying in a vacuum stripper under vacuum at above 500 mm of mercury to get dried products in powder form.

7. A method of producing the composition as claimed in claim 3 wherein the process steps comprising;

a) cleaning, drying, and powdering the roots of coconut tree,

b) carrying out extraction with methanol using an extractor preferably in a Soxhlet extractor for about 5 hrs at a temperature of about 65° C,

c) evaporating the resultant methanol extract preferably in a agitated thin film evaporator (ATFE) at a temperature of about 60 to 65°C to form concentrated methanol extract,

d) drying the resultant methanol extract under vacuum at above 500 mm of mercury to get dried product in powder form.

8. A method of producing the composition as claimed in claim 3, wherein the process step comprising:

a) cleaning, drying, and crushing the coconut shell,

b) extracting by adding with methanol in an amount ten times the quantity of crushed shells using an extractor with reflux condenser, to obtain a first residue and supernatant,

c) further extracting the first residue two more times with ten times the quantity of methanol at each time and separate the residue and supernatants d) pooling all the supernatants and concentrated in an agitated thin film evaporator (ATFE) at a temperature of about 60°C to form concentrated methanol extract,

e) drying the resultant concentrated methanol extract under vacuum at above 500 mm of mercury to get dried product in powder form.

9. A method of producing the composition as claimed in claim 3, wherein the process step comprising:

a) cleaning, drying, and crushing the coconut shell,

b) extracting by adding water in an amount ten times the quantity of crushed shells using an extractor with reflux condenser, to obtain a first residue and supernatant,

c) further extracting the first residue two more times with ten times the quantity of water at each time and separate the residue and supernatants.

d) pooling all the supernatants and concentrated in an agitated thin film evaporator (ATFE) at a temperature of about 85°C to form concentrated water extract.

e) drying the resultant concentrated water extract under vacuum at above 500 mm of mercury to get dried product in powder form.

10. A composition as claimed in claim 1 , having anti hyperglycemic and antioxidant activity is capable of using for the treatment of diabetes.

1 1 . A method of treating diabetes in mammals by the composition as claimed in claim 1 comprising of polyphenols in the range of 5-80%, flavonoids in the range of 0.2 - 10%; tannins in the range of 10- 60%; saponins in the range of 1 - 30%; alkaloids in the range of 1 -25%, anthocyanins in the range of 0.1 -5%, proteins in the range of 1 -10% and carbohydrate in the range of 1-30%.

12. A method of treating pre diabetes in mammals by the composition as claimed in claim 1 comprising of polyphenols in the range of 5-80%, flavonoids in the range of 0.2 - 10%; tannins in the range of 10- 60%; saponins in the range of 1 - 30%; alkaloids in the range of 1 -25%, anthocyanins in the range of 0.1 -5%, proteins in the range of 1 -10% and carbohydrate in the range of 1 -30%.

13. A method of reducing blood sugar level in mammals by the composition as claimed in claim 1 comprising of polyphenols in the range of 5-80%, flavonoids in the range of 0.2 - 10%; tannins in the range of 10- 60%; saponins in the range of 1 - 30%; alkaloids in the range of 1 -25%, anthocyanins in the range of 0.1 - 5%, proteins in the range of 1 -10% and carbohydrate in the range of 1 -30%.

14. A method of increasing the antioxidant activity in mammals by the composition as claimed in claim 1 comprising of polyphenols in the range of 5- 80%, flavonoids in the range of 0.2 - 10%; tannins in the range of 10- 60%; saponins in the range of 1 - 30%; alkaloids in the range of 1 -25%, anthocyanins in the range of 0.1-5%, proteins in the range of 1 -10% and carbohydrate in the range of 1 -30%.

15. A method of treatment as claimed in claims 1 1 to 14, wherein the composition is administered to patients in the form of oral dosage selected from the group consisting of a capsule, tablet, granule, sachet, powder, paste, ointment, solution, suspension, emulsion, in addition in the form of infusion, injection and ampoule and the daily dosage for treating a diabetic patient range from 250mg to 500mg, two to three times per day.

16. A method of treatment as claimed in claims 1 1 to 14, wherein the mammals include animals and human beings.

Description:
A COMPOSITION WITH ANTI HYPERGLYCEMIC AND ANTIOXIDANT ACTIVITY OBTAINED FROM THE EXTRACT OF PARTS OF COCONUT AND A METHOD OF PRODUCING THE SAME FIELD OF INVENTION

The present invention relates to an extract obtained from various parts of coconut tree like roots, trunk, leaves and also from shells, and husk of coconut also relates to a method of producing the said extract, and more specifically the extract enriched with polyphenol including flavonoids, tannins and also enriched with saponins, carbohydrates, anthocyanins, proteins and alkaloids for the treatment diabetes and to control the blood sugar level. This extract according to our invention is found to have anti hyperglycemic, anti diabetic and anti oxidant activity.

BACK GROUND OF THE INVENTION

Coconut, Cocos nucifera L., is a tree that is cultivated for its multiple utilities, mainly for its nutritional and medicinal values. The various products of coconut include tender coconut water, copra, coconut oil, raw kernel, coconut cake, coconut toddy, coconut shell and wood based products, coconut leaves, coir pith etc. It's all parts are used in some way or another in the daily life of the people in the traditional coconut growing areas. It is the unique source of natural products useful against various diseases and also for the development of industrial products. The coconut palm is, therefore, eulogized as ' alpavriksha' (the all giving tree) in Indian classics, and thus the current review describes the facts and phenomena related to its use in health and disease prevention. (Deb andal, M., 2010)

The coconut which is found throughout the tropic and subtropic area is known for its great versatility as seen in the many domestic, commercial, and industrial uses of its different parts. Some parts of coconut include coconut kernel, coconut roots, coconut trunk, coconut shell, coconut husk and coconut leaves. In tropical countries different parts of coconut are being used for their health benefits and it slowly got introduced to other civilization. It is also known as the "Tree of Life" due to the wide variety of products which can be derived from its various parts such as the use of Coconut tree products for building houses, decors, or health products that can be produce from its roots or coconut water. The edible parts of coconut fruit like coconut kernel and tender coconut water have numerous medicinal properties such as antibacterial, antifungal, antiviral, ant parasitic, antidermatophytic, antioxidant, hypoglycemic, hepatoprotective, immunostimulant etc. Coconut water and coconut kernel contain microminerals and nutrients, which are essential to human health, and hence coconut is used as food by the peoples in the globe, mainly in the tropical countries. The oil and milk derived from coconut kernel is commonly used in cooking and frying; coconut oil is also widely used in soaps and cosmetics.

Arginine, a coconut kernel protein (C P) has anti diabetic effect and is investigated on alloxan induced diabetes in Sprague-Dawley rats. CKP feeding attenuated the increase in the glucose and insulin level in diabetic rats. (Salil G et al, 201 1 ). Coconut water is reported to have inhibitory activity against human bacterial pathogens. (Mandal S et al, 2009). A warm water crude extract of coconut milk and coconut water dispersion has antiulcerogenic effects and were investigated in male Wistar albino rats. Results showed that coconut milk extract offered stronger protection against indomethacin-induced ulceration than coconut water in rats. (Nneli RO et al, 2008).

Majeed et al discloses the use of coconut water in mammalian tissue nourishment, growth and healthy maintenance. (Patent No.7300682). Reddy et al discloses an effective amount of herbal composition (extract of Cocos nucifera and other herbal extracts) applied topically to a human is suitable for regression of chronic inflammatory skin disease (Patent No.7666450). Rong-Tsun Wu discloses a method for treating immunological disease and / or disorders associated with tumor necrosis factor-alpha (TNF-∞) by administering effective amount of coconut water or shell in water.

At present coconut tree trunks and leaves and the coconut husks & shells are being used as material to make a variety of products for furnishing and decorating. It also has cultural and religious significance in many societies that use it. They can also be used for fuel and especially the coconut shells are a source of charcoal. Activated carbon manufactured from coconut shell is considered superior to those obtained from other sources, mainly because of small macropores structure which renders it more effective for the absorption of gas and vapor and for the removal of colour, oxidants, impurities and odor of compounds. Coconut trunks are used for building small bridges; they are preferred for their straightness, strength and salt resistance. It is also reported that the roots of coconut tree are used as a dye, a mouthwash, and a medicine for diarrheal and dysentery.

The medicinal properties of parts of coconut tree like roots, bark, trunk, leaves and coconut shell and husk are well known. The antibacterial effect of water extract of root and bark of Cocos nucifera Linn was reported. The study revealed that the root and bark of plant Cocos nucifera Linn, showed antibacterial activity against all the UTI isolates. ( . omala sivakumar, 201 1 ). The antimicrobial and antiviral activity of husk fiber from coconut was reported to be mainly due to high content of phenolic compounds. (Esquenazi D.et al.2002).The bioactivities of coconut also included inhibitory activity against acyclovir- resistant herpens simplex virus type 1 , leishmanicidal effects (Mendonca-Filho et al, 2004), protection of hemoglobin from nitrite-induced oxidation to methemoglobin (Mantena SK et al,2003), and free radical scavenging activities. Antihelmintic activity of the liquid extracted from the bark of the green coconut on intestinal nematodes of mice was studied. (Costa CT et al, 2010). An extract of Cocos-nucifera with catechin has found to be effective on lymphocytes proliferation. (Kirszberg C et al, 2003). Polyphenolic compounds from the aqueous extract of green coconut shell were isolated. Benzoyl ester derivatives were prepared with these polyphenols. Monobenzoyl and dibenzoyl derivatives of a polyphenol were separated and characterized. (Akhter, A et al, 2010). An alcoholic extract of coconut shell has found to be having antifungal activity (Venkataraman et al 1980).

But the use of parts of coconut tree like roots, bark, trunk, leaves and coconut shell and husk for hyperglycemia and for diabetes is not reported till now. But as part of our ongoing effort in finding new medicine for controlling diabetics, we have found that if we can make an extract from these items which is enriched with polyphenol including flavonoids, tannins and also enriched with saponins, carbohydrates, anthocyanin, protein and alkaloids which has anti hyperglycemic, anti diabetic and anti oxidant activity can be the right choice.

OBJECTIVE OF THE INVENTION

Therefore the main object of the present invention is to provide a medicine having anti hyperglycemic, anti diabetic and anti oxidant activity and also a method of producing the said medicine. Another object of the present invention is to produce the said medicine for the treatment for diabetics which is cheap, so that common man can afford.

Yet another objective of the present invention is to have said medicine which has no side effect.

The above mentioned objectives of the present invention have been achieved based on our findings by identifying the most efficient method of isolating extract enriched with polyphenol including flavonoids, and tannins, and also enriched with saponins, carbohydrates, anthocyanins, proteins and alkaloids from parts of coconut tree like roots, bark, trunk, leaves and coconut shell and husk which is having anti hyperglycemic, anti diabetic and anti oxidant activity.

SUMMARY OF THE INVENTION:

The present invention provides a composition having anti hyperglycemic, antidiabetic and antioxidant activity which can be obtained from the extract of parts of coconut tree selected from roots, bark, trunk, leaves and coconut shell and husk or combinations thereof comprising enriched polyphenol including flavonoids and tannins and also enriched with saponins, carbohydrates, anthocyanins, proteins and alkaloids wherein polyphenols are present in the extract in a range of 5% and above, flavonoids are present in the extract in a range of 0.2% and above, tannins are present in the range of 10% and above, saponins are present in the range of 1 % and above, alkaloids are present in the range of 1 % and above, anthocyanins are present in the range of 0.1 % and above, proteins are present in a range of 1 % and above and carbohydrate are present in the range of 1 % and above.

The method of producing the said composition comprises the steps of; a) selecting the raw material and cleaning to make it free of any foreign mater like dirt, b) drying the cleaned material at a temperature ranging from 100 tol 50 °C, c) grinding or crushing the dried material into powder form, d) by extraction with · solvents selected from water, hexane, methanol, ethanol, isopropanol, n-butanol, methyl acetate, ethyl acetate, propyl acetate, n- butyl acetate and combinations thereof to obtain an extract enriched with polyphenol including flavonoids and tannins and also enriched with saponins, carbohydrates, anthocyanins, proteins and alkaloids, e) concentrating and drying the resultant extract to obtain in dry powder form, Said extract enriched with polyphenol including flavonoids, and tannins and also enriched with saponins, carbohydrates, anthocyanins, proteins and alkaloids obtained from parts of coconut tree like roots, bark, trunk, leaves and coconut shell and husk and has the property of scavenging the free radicals as well.

In an embodiment under the present invention said extract enriched with polyphenols including flavonoids and tannins and also enriched with saponins, carbohydrates, anthocyanins, proteins and alkaloids, can be prepared by extraction of coconut root with water and further purified by column chromatography.

In another embodiment said extract is " prepared by successive extraction of water extract of coconut root by using hexane, ethyl acetate and butanol.

In yet another embodiment said extract is prepared from root of coconut by extraction with alcohol and water.

Another embodiment provides the preparation of the said extract by the extraction of the coconut shell with water.

Yet another embodiment provides the preparation of the said extract by the extraction of the coconut shell with alcohol.

The disclosure also provides a ' dosage form the above mentioned extract derived from the parts of coconut tree like roots, bark, trunk, leaves and coconut shell and husk. The dosage is can be oral administration and is in the form selected from the group consisting of a capsule, tablet, granule, sachet, powder, paste, ointment, solution, suspension, emulsion, etc. It can also be in the form of infusion, injection, ampoule etc,

BRIEF DESCRIPTION OF THE DRAWINGS

The Invention is described with reference to figures 1 to 6 of the drawings.

Fig 1 provides anti-oxidant activity of water extract of coconut root by DPPH ( 1 , 1 - diphenyl-2-picrylhydrazyl radical) method

Fig 2 provides anti-oxidant activity of methanol extract of coconut root by DPPH (1 , 1 - diphenyl-2-picrylhydrazyl radical) method

Fig 3 provides anti-oxidant activity of methanol extract of coconut shell by DPPH (1 , 1 - diphenyl-2-picrylhydrazyl radical) method

Fig 4 provides anti-oxidant activity of aqueous extract of coconut shell by DPPH ( 1 , 1 - diphenyl-2-picrylhydrazyl radical) method Fig 5 provides anti-oxidant activity of ethyl acetate extract of water extract of coconut root by DPPH (1 , l -diphenyl-2-picrylhydrazyl radical) method

Fig 6 provides anti-oxidant activity of methanol elute of column purified water extract of coconut root by DPPH (1 , 1 -diphenyl-2-picrylhydrazyl radical) method

DETAILED DESCRIPTION OF THE INVENTION

Other features and advantages of the present invention should become apparent from the following description of the preferred embodiment under this invention read in conjunction with the accompanying drawings, which illustrate, by way of example, the principles of the invention.

The present invention relates to an extract enriched with polyphenol including flavonoids and tannins and also enriched with saponins, carbohydrates, anthocyanins, proteins and alkaloids obtained from parts of coconut tree like roots, bark, trunk, leaves and coconut shell and husk useful for reducing blood sugar level and for the treatment of diabetes. According to a preferred embodiment under the invention the method of producing an extract of coconut enriched with polyphenol including flavonoids, and tannins and also enriched with saponins, carbohydrates, anthocyanins, proteins and alkaloids, is from the root of coconut tree. The extract of coconut obtained from the root of coconut having anti hyperglycemic, anti diabetic and anti oxidant activity can be further purified by column chromatography to enhance the anti hyperglycemic, anti diabetic and anti oxidant activity. Similar extract can be made from trunks, leaves of coconut tree as well as from coconut shell & husk by similar method as followed in the case of root.

It is further noted that said extract of coconut root enriched with polyphenol including flavonoids and tannins and also enriched with saponins, carbohydrates, anthocyanins, proteins and alkaloids is having the property of scavenging the free radicals. Free radicals are atoms or group of atoms that can cause damage to cells, impairing the immune system and leading to infections and various degenerative diseases such as heart disease and cancer. Free radicals are thought to be a by product from a number of causes including environmental factors, unhealthy food, smoking, radiation and a number of other sources. Antioxidants are molecules which can safely interact with free radicals and terminate the chain reaction before vital molecules are damaged. Free radical scavenging activity is determined by using 1 , l -diphenyl-2-picrylhydrazyl (DPPH ) free radical. The DPPH assay method is based on the reduction of DPPH, a stable free radical. The effect of the extract under this invention on fasting blood glucose level in streptozotocin and alloxan induced diabetic rats was also studied. A significant reduction in fasting blood glucose level is observed after the administration the extract. It was also noticed that extract of roots of coconut tree enriched with polyphenol including flavonoids and tannins and also enriched with saponins carbohydrates, anthocyanins, proteins and alkaloids when administered in patients with impending diabetes and diabetes caused a decrease in fasting blood sugar and post prandial blood glucose levels.

The present invention provides a composition obtained from the extract of parts of coconut tree selected from roots, bark, trunk, leaves and coconut shell and husk or combinations thereof comprising enriched polyphenol including flavonoids and tannins and also enriched with saponins, carbohydrates, anthocyanins, proteins and alkaloids wherein polyphenols are present in the extract in arrange of 5-80%, flavonoids are present in the extract in a range of 0.2 - 10%, tannins are present in the range of 10- 60%, saponins are present in the range of 1- 30%, alkaloids are present in the range of 1 -25%, anthocyanins are present in the extract in a range of 0.1- 5%, proteins are present in a range of 1 % - 10% and carbohydrate are present in the range of 1 -30%.

One preferred embodiment provides the method of preparing water extract of obtained from different parts of coconut tree like roots, bark, trunk, leaves and coconut shell and husk preferably roots of coconut tree. The extract can be further purified by column chromatography resulting in to dried powder of methanol extract of purified water extract. In another embodiment by extraction with water and then successive extraction with solvents selected from hexane, methanol, ethanol, isopropanol, n-butanol, methyl acetate, ethyl acetate, propyl acetate, n-butyl acetate and combinations thereof can also result into similar extract from different parts of coconut tree like roots, bark, trunk, leaves and coconut shell and husk preferably roots of coconut tree and the resultant dried powder of the extract has the anti hyperglycemic, anti diabetic and anti oxidant activity .

The disclosure provides a dosage form of the said extract for oral administration. Oral dosage forms of the extract are selected from the group consisting of a capsule, tablet, granule, sachet, powder, paste, ointment, suspension, emulsion, pills etc and it can also administered in the form of infusion, injection, ampoule and the daily dosage for treating a diabetic patient range from 250mg to 500mg, two to three times per day. It has been observed that a dosage form of an extract of roots of coconut tree and coconut shell administered in a dosage of 50mg/Kg/day to streptozotocin-induced diabetic rats is very effective. It has also been observed that a dosage form of an extract of roots of coconut tree and coconut shell administered in a dosage of 50mg/K.g/day to alloxan- induced diabetic rats is very effective.

Further a dosage form of an extract of coconut root administered in a dosage of 500mg capsule /day to patients with impending diabetes and diabetes has resulted encouraging results in controlling the blood sugar level. Hence the present invention can be a great step forward in for treating animals and human beings suffering from diabetes.

Various methods for the preparation of extract enriched with polyphenol, including flavonoids and tannins and also enriched with saponins carbohydrates, anthocyanins, proteins and alkaloids prepared by the extraction of roots of coconut tree are as under:

1. Extraction of roots of coconut tree first with water and further purified by column chromatography and resulting in to water elute and methanol elute which are further dried. But the resultant dried powder of methanol extract of purified water extract is found to be superior.

2. Extraction of roots of coconut with water and then successive extraction with hexane, ethyl acetate and butanol and resulting into a) dried powder of hexane extract b) dried powder of ethyl acetate c) Powder of butanol extract. But the dried powder of ethyl acetate is found to be of high order.

3. Extraction of coconut root with water and resultant dried Powder of water extract.

4. Extraction of coconut root with alcohol (methanol) and resultant Powder of methanol extract.

Various methods for the preparation of extract enriched with polyphenol including flavonoids and tannins and also enriched with saponins carbohydrates, anthocyanins, proteins and alkaloids prepared by the extraction of coconut shell are as under:

1 . Extraction of coconut shell with water and resultant dry powder of water extract

2. Extraction of coconut shell with alcohol (methanol) and dry powder of methanol extract

All the above extracts found to have anti hyperglycemic, anti diabetic and anti oxidant activity. One of the preferred embodiments mentioned above provides the method of preparing water extract of coconut root.

[3] Roots of coconut tree are dried and powdered. Powdered roots are extracted with water. The mixture is refluxed for one hour to obtain a first residue and supernatant. After first extraction, the first residue is further extracted two more times with ten times the quantity of water at each time. The residue and supernatants are separated. All the supernatants are pooled and concentrated. Concentrated supernatant is dried to get water extract of roots of coconut.

[ 1 ] In yet another preferred embodiment provides method of preparation of purified water extract of roots of coconut tree. Water extract of coconut root is passed through HP20 column (Chromatographic method of purification) and eluted with water and methanol. Each part collected is concentrated and dried separately to form purified water extract of coconut root. But the resultant dried powder of methanol extract of purified water extract is found to be superior.

[4] In another embodiment under the invention the method of preparing methanol extract of coconut root is as under. Coconut roots are dried and powdered. Powdered roots are extracted with methanol in a Soxhiet extractor. The extraction is carried out for 5hrs. After completion of extraction, the solvent is filtered and concentrated to remove the methanol. Concentrated methanol extract is dried to get methanol extract of coconut root.

[2] In yet another embodiment under the invention extract of roots of coconut tree is prepared by successive extraction using solvents. Coconut roots are dried and powdered. Powdered roots are extracted with water to form powder of water extract of coconut root. Powder of water extract of coconut root is dissolved in water which can be extracted by adding hexane and subsequent separation of the two phases -aqueous and hexane. Collect the hexane phase and ethyl acetate is added to the aqueous phase. Shake the aqueous and ethyl acetate phase well and subsequent separation of the two phases -aqueous and ethyl acetate. Collect the ethyl acetate phase and butanol is added to the aqueous phase. Shake well and subsequent separation of the two phases -aqueous and butanol and collect the butanol part. Each collected part is concentrated and dried separately. But the dried powder of ethyl acetate is found to be of high order.

[5] Another embodiment provides the method of preparing methanol extract from coconut shell. Shells of coconut are crushed and then extracted with methanol. The mixture is refluxed for one hour to obtain a first residue and supernatant. After first extraction, the first residue is further extracted two more times with ten times the quantity of methanol at each time. The residue and supernatant are separated. All the supernatants are pooled and concentrated. Concentrated supernatant is dried to get methanol extract of coconut shell.

[6] Still another embodiment provides the method of preparing water extract of coconut shell. Shells of coconut are crushed extracted with water. The mixture is refluxed for one hour to obtain a first residue and supernatant. After first extraction, the first residue is further extracted two more tipes with ten times the quantity of water at each time. The residue and supernatants are separated. All the supernatants are pooled and concentrated. Concentrated supernatant is dried to get water extract of shell of coconut.

The extract obtained as disclosed in the present invention obtained from the parts of roots of coconut as mentioned earlier are useful for reducing blood sugar level and to treat diabetes by administering extract to a mammal. Mammals include animals and human beings.

Example - 1 (Extraction from root with water)

Roots of coconut were collected (100 Kg). Roots were cleaned and dried. Dried roots were powdered. Water in an amount ten times the quantity of powder of coconut root was added to the powder of coconut to form a mixture. The extraction was performed using an extractor with a reflux condenser. The bottom of the extractor was fitted with a polypropylene (100 microns) filter cloth. The mixture was refluxed for one hour to obtain a first residue and supernatant. After the first extraction, the first residue was further extracted two more times with ten times the quantity of water at each time. The residue and supernatants were separated by draining out the supernatant from the extractor bottom through the polypropylene filter cloth using a centrifugal pump. All the supernatants were pooled and concentrated in an Agitated thin film evaporator (ATFE) at a temperature of 85°C to form concentrated water extract. Concentrated water extract was dried under vacuum at above 500 mm of mercury to get powder of water extract of roots of coconut (Yield l OKg).

Polyphenol content : 127mg/g of extract (UV method)

Flavonoid content : 18mg/g of extract (Gravimetric method)

Tannin content : 227.4 mg/g of extract (UV method) Saponin content : 70mg/g of extract (Gravimetric method)

Alkaloid content : 46.8mg/g of extract (Gravimetric method)

Carbohydrate content: 76.1 mg/g of extract (UV method)

Anthocyanin content: 1 mg/g of extract ((Gravimetric method)

Protein content : 20mg/g of extract ((UV method)

Antioxidant activity of the powder of water extract of coconut root by DPPH (1 , 1 - diphenyl-2-picrylhydrazyl radical) method was found to be 1

Example 2 (Extraction from root with water to get more purified product)

Roots of coconut were collected (100 Kg). Roots were cleaned and dried. Dried roots were powdered. Water in an amount ten times the quantity of powder of coconut root was added to the powder of coconut to form a mixture. The extraction was performed using an extractor with a reflux condenser. The bottom of the extractor was fitted with a polypropylene (100 microns) filter cloth. The mixture was refluxed for one hour to obtain a first residue and supernatant. After the first extraction, the first residue was further extracted two more times with ten times the quantity of water at each time. The residue and supernatants were separated by draining out the supernatant from the extractor bottom through the polypropylene filter cloth using a centrifugal pump. All the supernatants were pooled and concentrated in an Agitated thin film evaporator (ATFE) at a temperature of 85°C to form concentrated water extract. Concentrated water extract was dried under vacuum at above 500 mm of mercury to get powder of water extract of roots of coconut (Yield l OKg).

Powder of water extract of roots of coconut was dissolved in water and centrifuged at 10,000 rpm for 10 minutes to form supernatant and residue. Supernatant was loaded on the HP20 column. The column was washed with water and water part was collected. After the water wash, column was washed with methanol and methanol part was collected. Each collected part was separately concentrated in an Agitated thin film evaporator (ATFE) to form concentrated extract of each part. Concentrate of each part was fed into vacuum stripper and dried separately under vacuum at above 500 mm of mercury to form purified water extract of coconut root.

Polyphenol content in methanol part= 600mg/g of extract (UV method)

Flavonoid content: 63mg/g of extract (Gravimetric method) Tannin content: 396.8mg/g of extract (UV method)

Saponin content: 120mg/g of extract (Gravimetric method)

Alkaloid content: 86.5mg/g of extract (Gravimetric method)

Carbohydrate content: 145mg/g of extract (UV method)

Anthocyanin content: 4mg/g of extract ((Gravimetric method)

Protein content : 32mg/g of extract ((UV method)

Antioxidant activity of the powder of methanol extract of purified water extract of coconut root was found by DPPH (1 , 1 -diphenyl-2-picrylhydrazyl radical) method.

Antioxidant activity by DPPH method = 2.7 μg/ml

Example 3 (Successive extraction of water extract of coconut root)

Roots of coconut were collected (100 Kg). Roots were cleaned and dried. Dried roots were powdered. Water in an amount ten times the quantity of powder of coconut root was added to the powder of coconut to form a mixture. The extraction was performed using an extractor with a reflux condenser. The bottom of the extractor was fitted with a polypropylene (100 microns) filter cloth. The mixture was refluxed for one hour to obtain a first residue and supernatant. After the first extraction, the first residue was further extracted two more times with ten times the quantity of water at each time. The residue and supernatants were separated by draining out the supernatant from the extractor bottom through the polypropylene filter cloth using a centrifugal pump. All the supernatants were pooled and concentrated in an Agitated thin film evaporator (ATFE) at a temperature of 85°C to form concentrated water extract. Concentrated water extract was dried under vacuum at above 500 mm of mercury to get powder of water extract of roots of coconut (Yield l OKg).

• Powder of water extract of roots of coconut is dissolved in water and transferred into a liquid-liquid extractor and extracted with hexane. Hexane phase and aqueous phase were separated.

• Hexane part was collected and ethyl acetate was added to the aqueous phase in the liquid-liquid extractor and subsequent separation of the two phases -aqueous and ethyl acetate. • After extraction ethyl acetate part was collected and butanol was added to aqueous phase in the liquid -liquid extractor and subsequent separation of the two phases - aqueous and butanol.

• After extraction butanol part was collected.

Each collected part was separately concentrated in an Agitated thin film evaporator (ATFE) to form concentrated extract of each part. Concentrate was fed into vacuum stripper and dried under vacuum at above 500 mm of mercury to form hexane (0.005 g), ethyl acetate (1 .2 Kg) and butanol (2Kg) extract of roots of coconut.

Polyphenol content in ethyl acetate extract of coconut root: 330mg/g of extract (UV method)

Flavonoid content: 39mg/g of extract (Gravimetric method)

Tannin content: 286.5mg/g of extract (UV method)

Saponin content: 93 mg/g of extract (Gravimetric method)

Alkaloid content: 61 .4 mg/g of extract (Gravimetric method)

Carbohydrate content: 105 mg/g of extract (UV method)

Anthocyanin content: 2.3 mg/g of extract ((Gravimetric method)

Protein content : 26mg/g of extract ((UV method) Polyphenol content in butanol extract of coconut root: 10.5 %( UV method)

Antioxidant activity of the powder of ethyl acetate extract of water extract of coconut root by DPPH (1 , l -diphenyl-2-picrylhydrazyl radical) method was found to be 5.5966 μg/ml. Antioxidant activity of the powder of butanol extract of water extract of coconut root by DPPH ( 1 , l -diphenyl-2-picrylhydrazyl radical) method was found to be 13.5 μg/ml.

Example 4 (Extraction from root with methanol)

Optional extraction method of coconut root is extraction of coconut root with methanol Roots of coconut were collected (100 Kg). Roots were cleaned and dried. Dried roots were powdered. Powdered roots were filled in the Soxhlet apparatus and extracted with methanol (400L) .The extraction is carried out for 5 hrs at a temperature of about 65° C. After the completion of extraction, the supernatant was filtered and concentrated in an Agitated thin film evaporator (ATFE) at a temperature of 60°C to form concentrated methanol extract. Concentrated methanol extract was dried under vacuum at above 500 mm of mercury to get methanol extract of roots of coconut (Yield 8Kg).

Polyphenol content: 12.9 %( UV method)

Antioxidant activity of the powder of methanol extract of coconut root by DPPH ( 1 , 1 - diphenyl-2-picrylhydrazyl radical) method was found to be 12.04μg/ml.

Example 5 (Extraction from shell with Methanol)

Optional extraction method of shell is extraction of coconut shell with methanol

Shells of coconut were collected (100 Kg). Shells were crushed into small pieces. Methanol in an amount ten times the quantity of crushed shells was added for methanol extraction. The extraction was performed using an extractor with reflux condenser. The bottom of the extractor was fitted with a polypropylene (100 microns) filter cloth. The mixture was refluxed for one hour to obtain a first residue and supernatant. After first extraction, the first residue was further extracted two more times with ten times the quantity of methanol at each time. The residue and supernatants were separated by draining out the supernatant from the extractor bottom through the polypropylene filter cloth using a centrifugal pump. All the supernatants were pooled and concentrated in an Agitated thin film evaporator (ATFE) at a temperature of 60°C to form concentrated methanol extract. Concentrated methanol extract was dried under vacuum at above 500 mm of mercury to get methanol extract of shell of coconut (Yield 4 Kg).

Polyphenol content: 30 %( UV method)

Antioxidant activity of the powder of methanol extract of coconut shell by DPPH (1 , 1 - diphenyl-2-picrylhydrazyl radical) niethod was found to be 9.2833 μg/m I. Example 6 (Extraction from shell with water)

Optional extraction method of coconut shell is extraction of coconut shell with water. Shells of coconut were collected (100 Kg). Shells were crushed into small pieces. Water in an amount ten times the quantity of crushed shells was added for water extraction. The extraction was performed using an extractor with reflux condenser. The bottom of the extractor was fitted with a polypropylene (100 microns) filter cloth. The mixture was refluxed for one hour to obtain a first residue and supernatant. After first extraction, the first residue was further extracted two more times with ten times the quantity of water at each time. The residue and supernatants were separated by draining out the supernatant from the extractor bottom through the polypropylene filter cloth using a centrifugal pump. All the supematants were pooled and concentrated in an Agitated thin film evaporator (ATFE) at a temperature of 85°C to form concentrated water extract. Concentrated water extract was dried under vacuum at above 500 mm of mercury to get water extract of shell of coconut (Yield 1.8 Kg).

Polyphenol content: 25.66 %( UV method)

Antioxidant activity of the powder of water extract of coconut shell by DPPH (1 , 1 - diphenyl-2-picrylhydrazyl radical) method was found to be 15.489μg/ml. Example 7 ( Determination of anti-oxidant property of methanol and water extract of coconut root and shell and ethyl acetate extract of water extract of coconut root and methanol extract of column purified water extract of coconut root by DPPH(1, 1- diphenyl-2-picrylhydrazyl radical) method)

The powdered sample (methanol or water extract of coconut root or shell, ethyl acetate extract of water extract of coconut root, methanol extract of column purified water extract of coconut root) was diluted with methanol to prepare a series of five concentrations ranging from 0 to 250μg/ml. To each of 2.9ml of the dilution taken in a spectrophotometer cuvette was added 0.1 ml of O.l mM methanolic DPPH solution. The contents were mixed, incubated in dark for 20min. and absorbance measured at 517 nm. A graph was constructed with concentration of the extractives in the reaction medium on the x-axis and percentage DPPH quenched on the y-axis. From the relationship, the concentration of extractives resulting in 50% quenching of DPPH was worked out and expressed as IC50. [Ref: Huang et al (2005)]

a) Determination of anti-oxidant property of water extract of coconut root by DPPH method

EC 50 The concentration of sample required for 50% quenching of free radical EC 50 = 50/ Intercept

Intercept = 4.181 (from graph)

Antioxidant activity of water extract of coconut root by DPPH method, EC 50 = 1 1.95 Γη 1.

b) Determination of anti-oxidant property of methanol extract of coconut root by DPPH method

EC 50 The concentration of sample required for 50% quenching of free radical

EC 50 = 50/ Intercept

Intercept = 4.151 (from graph)

Antioxidant activity of methanol extract of coconut root by DPPH method, EC 50 = 12.04 μ^Γη Ι .

c) Determination of anti-oxidant property of methanol extract of coconut shell by DPPH method

EC 50 The concentration of sample required for 50% quenching of free radical

EC 50 = 50/ Intercept

Intercept = 5.386 (from graph) Antioxidant activity of methanol extract of coconut shell by DPPH method, EC 50 = 9.2833 μ^πιΐ.

d) Determination of anti-oxidant property of water extract of coconut shell by DPPH method

EC 50 The concentration of sample required for 50% quenching of free radical

EC 50 = 50/ Intercept

Intercept = 3.228 (from graph)

Antioxidant activity of water extract of coconut shell by DPPH method, EC 50 =15.489μg/mI

e) Determination of anti-oxidant property of ethyj acetate extract of water extract of coconut root by DPPH method

EC 50 The concentration of sample required for 50% quenching of free radical

EC 50 = 50/ Intercept

Intercept = 8.934 (from graph)

Antioxidant activity of ethyl acetate extract of water extract of coconut root by DPPH method, EC 50 =5.5966 μg /ml f) Determination of anti-oxidant property of methanol extract of column purified water extract of coconut root by DPPH method

EC 50 The concentration of sample required for 50% quenching of free radical

EC 50 = 50/ Intercept

Intercept = 1 8.38 (from graph)

Antioxidant activity of methanol part of column purified water extract of coconut root by DPPH method, EC 50 =2.7 μg /ml Example 8 (Method of analysis of polyphenol by UV method)

Polyphenol was estimated by a spectrophotometric method. Ferrous tartrate solution was prepared by weighing 0.1 g of ferrous sulfate and 5 gm of potassium sodium tartrate into a 1000ml volumetric flask and made up to the mark using water.

Phosphate buffer solution(PH7.5) was prepared by weighing 23.377 g of disodium hydrogen phosphate and dissolved in water and made up to 1000ml using water(A).9.078 g of potassium dihydrogen phosphate was weighed and dissolved in water and made up to 1000ml using water(B)

85ml of solution A and 15ml of solution B was mixed to get phosphate buffer solution. 0.1 g of the extract was weighed and dissolved in water and made up to 100ml using water.

1 ml of the test solution (sample) was taken in a 25 ml measuring flask and 4 ml water was added. Then 5 ml of ferrous tartrate solution was added and shake well. Then made up with phosphate buffer solution and shake well. Absorbance was noted at 540 nm with water as blank. The percentage of polyphenols can be calculated using the formula: Polyphenols % = 2.884x A x L, x 100%

1000 x L 2 W

Where

W - Weight of test sample (g)

Li - volume of test solution (ml)

L 2 _ volume of test solution injected in the spectrophotometer (ml)

A - Absorbance values

Example 9(Method of analysis of carbohydrate)

Carbohydrate was estimated by a spectrophotometric method using anthrone reagent as described in David T Plummer - An introduction to practical biochemistry: p-183. Standard was prepared by weighing 0.02 g of glucose, and made up to 100ml with water. The sample was made up by taking 0.05 g of dry extract of in 50 ml water. Anthrone reagent was prepared by taking 0.2 g anthrone in 100 ml concentrated H2SO4.

0.1 ml of the test solution (sample & standard) was taken in a 10 ml stoppered test tube and the solution was made up to 1 ml with distilled water (0.1 ml of test solution and 0.9 ml of distilled water). The tubes were kept on ice. 0.4 ml of anthrone reagent was added to the above solution and mixed rapidly. The tubes were closed and placed in boiling water bath for 10 minutes. Absorbance was noted at 620 nm after cooling the solution. By comparing the absorbance and concentration of standard and sample, the percentage of carbohydrate can be calculated using the formula:

Absorbance of sample X Concentration of standard

Absorbance of standard X Concentration of sample

Example 10(Method of analysis of Flavonoids)

Flavonoids was determined by gravimetric method as described by Harborne (1973).The sample( 5 g) was boiled in 50ml of 2M hydrochloric acid solution for 30 minutes under reflux. It was allowed to cool and then filtered through whatman No 42 filter paper .A measured volume of the extract was treated with equal volume of ethyl acetate starting with drop. The flavonoid precipitated was recovered by filtration using weighed filter paper. The resulting weight difference gave the weight of flavonoid in the sample. Example 1 1 (Method of analysis of Saponins)

Saponins were estimated by gravimetric method (Obadoni and Ochuko, 2001 ).

The sample was ground and 20mg of sample was put into a conical flask and 100cm 3 of aqueous ethanol were added. The samples were heated over a water bath for 4h with continuous stirring at about 55°C. The mixture was filtered and the residue was re- extracted with another 200ml 20% ethanol. The combined extracts were reduced to 40ml over water bath at about 90°C. The concentrate was transferred into a 250ml separatory funnel and 20ml of ethyl ether was added and shaken vigorously. The aqueous layer was recovered while the ether layer was discarded. The purification process was repeated. 60ml of n-butanol was added. The combined n-butanol extracts were washed twice with 10ml of aqueous sodium chloride. The remaining solution was heated in a water bath. After evaporation the sample were dried in oven in weighed filter paper. After drying the filter paper with dried material was weighed. The weight difference of empty filter paper and filter paper with residue gave the percentage of saponins present in the sample of dry extract of coconut.

Example 12(method of analysis of alkaloid)

Alkaloid was estimated by gravimetric method using Harborne (1973) method. 5g of the sample was weighed into 250ml beaker and 200ml of 10% acetic acid in ethanol was added, covered and allowed to stand for 4h. This was filtered and the extract was concentrated on a water bath to one-quarter of the original volume. Concentrated ammonium hydroxide was added drop wise to the extract until the precipitation was complete. The whole solution was allowed to settle and the precipitated was collected and washed with dilute ammonium hydroxide and then filtered through a weighed filter paper The residue was the alkaloid, which was dried and weighed. The weight difference of empty filter paper and filter paper with residue gave the percentage of alkaloid present in the sample of dry extract of coconut.

Example 13(method of estimation of tannins)

Tannin content was determined by the Folis-Denis colorimetric method described by Kirk and Sawyer (1998). 5g of the sample was weighed into a 50ml plastic bottle. 50ml of distilled water was added and shaken in a mechanical shaker. The mixture was allowed to stand for 30min at 28°C. After 30 minutes the mixture was filtered through what man No.

42 grade of filter paper to get a filtrate. 2ml of the filtrate was pipetted into a 50ml volumetric flask. Similarly 2ml standard tannin solution (20mg tannic acid in 50 ml volumetric flask made up with distilled water) and 2ml of distilled water(blank) were put in separate volumetric flasks and 2.5ml of saturated sodium carbonate(Na 2 C03) solution added to each of the flasks. The content of each flask was made up to 50mls with distilled water and allowed to incubate at 28°C for 90 min. Their respective absorbance was measured in a spectrophotometer at 260nm.

Absorbance of sample X Concentration of standard

Absorbance of standard X Concentration of sample

Example 14(method of estimation of anthocyanin)

Anthocyanin was estimated gravimetrically by the method of Harborne (1973). 5g of the sample was weighed and hydrolyzed by boiling in 100ml or 2M hydrochloric acid solution for 30 min. The hydrolysate was filtered using whatman No 42 filter paper. The filtrate was transferred into a separation funnel and equal volume of ethyl acetate was added to it, mixed well and allowed to separate into two layers. The ethyl acetate layer (extract) was collected while the aqueous layer was discarded. The extract was separated to dryness in the crucible over a steam bath. The dried extract was then treated with concentrated amyl alcohol to extract the anthocyanins. After filtration, the alcohol extract and the filtrate was transferred to a weighed evaporating dish and evaporated to dryness. It was then dried in the oven at 300C for30min and cooled in a desiccator. The weight of anthocynanin was determined and expressed as percentage of the original sample.

Example 15(method of estimation of protein)

Total protein in the given sample is estimated by spectrophotometric method. 50mg of the sample is made upto 50ml with 0.1 N NaOH. l Omg/l Oml standard bovine serum albumin is prepared using 0.1N NaOH. Different volumes of standard and sample solution (0.55ml-0.25ml) were taken and made up to 0.55 ml using distilled water. 2.5ml of fresh alkaline copper reagent solution (Fresh copper reagent is prepared by mixing one part of copper sulphate (l %w/v), one part of sodium potassium tartarate (2%w/v). To one volume of the above mixture, 50 volume of 2 %( w/v) sodium carbonate in 0.1N sodium hydroxide was added) was added to the samples and incubated for 15 minutes at room temperature. 0.25ml of freshly constituted Follins phenol reagent was added to the sample and standard solutions and incubated for 30 minutes at room temperature. The absorbance is read at 660nm in spectrophotometer. By comparing the absorbance and concentration of standard and sample, the percentage of protein can be calculated using the formula:

Absorbance of sample X Concentration of standard x 100

Absorbance of standard X Concentration of sample

Example 16 (Method of preparation of 500 nig capsule of coconut extract)

A 500 mg capsule containing 500 mg of a powder of extract is prepared by encapsulating the powder in hard 0 size gelatin capsules. The process is performed in an air-conditioned at 21 °C and de- humidified room. 2 kg of powder is charged into the hopper of a semiautomatic capsule filling machine. '0' size hard gelatin capsule shell is loaded to the tray and the extract powder is filled into the shell. The filled weight of capsules are checked simultaneously and these capsules are sorted by a sorting machine and polished with the help of a polishing machine to give 4000 capsules of 500 mg each.

Example 17 (Effect of powder of coconut root extract and coconut shell extract on blood glucose level in streptozotocin-induced diabetic rats)

The study was conducted to evaluate the effect of powder of coconut root and coconut shell extracts. The study was conducted in 30 adult male Sprague-Dawley rats weighing

200-250gm. The animals were randomly divided into five groups comprising six each.

Diabetes was induced in all the groups except the normal control group by intra peritoneal administration of Streptozotocin at 50 mg/kg body weight. Blood glucose levels were estimated on initial day and on day 5, to confirm the induction of diabetes. After confirming the induction of diabetes, except the diabetic control group, test drugs (extract being tested or standard drug) was administered to each group for 28 days. The fasting blood glucose level (FBG) was assessed on day 3, day 7, day 14, day 21 and day 28. The experimental design was as follows:

Group I - Normal control

Group II - Diabetic control

Group III - Diabetic rats treated with glibenclamide 1 mg/kg body weight orally for 28 days Group IV - Diabetic rats treated with powder of alcoholic extract of coconut root at 50mg/kg body weight orally for 28 days

Group V - Diabetic rats treated with powder of aqueous extract of coconut shell at 50mg/kg body weight orally for 28 days

Observations:

Fasting blood glucose levels before induction of diabetes, after induction of diabetes (day

0) and on days 3, 7, 14, 21 and 28

Results:

1 . Fasting blood glucose levels (FBG)

The results of effect of powder of alcoholic extract of coconut root and aqueous extract of coconut shell on fasting blood glucose levels are presented in Table.1 . FBG levels of Groups III, IV and V showed significant reduction with 28 days of test drug administration.

Table 1 . Effect of powder of alcoholic extract of coconut root and aqueous extract of coconut shell extract on fasting blood glucose levels in streptozotocin-induced diabetic rats. Data is reported as ean±SE , n=6

Example 18 (Effect of powder of coconut root extract and coconut shell extract on blood glucose level in alloxan-induced diabetic rats)

The study was conducted to evaluate the effect of powder of coconut root and coconut shell extracts. The study was conducted in 30 adult male Sprague-Dawley rats weighing 200-250gm. The animals were randomly divided into five groups comprising six each. Diabetes was induced in all the groups except the normal control group by intra peritoneal administration of alloxan at 50 mg/kg body weight. Blood glucose levels were estimated on initial day and on day 5, to confirm the induction of diabetes. After confirming the induction of diabetes, except the diabetic control group, test drugs (extract being tested or standard drug) was administered to each group for 28 days. The fasting blood glucose level (FBG) was assessed on day 3, day 7, day 14, day 21 and day 28. The experimental design was as follows:

Group 1 - Normal control

Group II - Diabetic control

Group III - Diabetic rats treated with glibenclamide I mg/kg body weight) orally for 28 days

Group IV - Diabetic rats treated with powder of aqueous extract of coconut root at 50mg/kg body weight orally for 28 days

Group V - Diabetic rats treated with powder of alcoholic extract of coconut shell at 50mg/kg body weight orally for 28 days

Results:

Fasting blood glucose levels (FBG)

The results of effect of powder of aqueous extract of coconut root and alcoholic extract of coconut shell on fasting blood glucose levels in alloxan-induced diabetic rats are presented in Table.2 FBG levels of Groups III, IV and V showed significant reduction with 28 days of test drug administration.

Table 2. Effect of powder of aqueous extract of coconut root and alcoholic extract of coconut shell extract on fasting blood glucose levels in alloxan-induced diabetic rats.

Data is reported as Mean±SEM, n=6

Example 19 (Effect of methanol extract of column purified water extract of coconut root on fasting blood sugar levels (FBS) and post prandial blood sugar levels (PPBS) in patients with impending diabetes)

Patients of either sex, aged between 40 and 60 years, and without severe systemic diseases were screened for the study. Among them, 18 patients with FBS: 100-120 mg/dl and PPBS: 140-190 mg/dl who were, not grossly obese and who gave informed consent were randomly selected and randomized into two groups. A prescribed diet was advised for the patients throughout the study period.

Group I - patients treated with glibenclamide 5 mg tablet once daily before breakfast for 4 weeks.

Group II - patients treated with methanol extract of column purified water extract of coconut root 500 mg capsule once daily before breakfast for 4 weeks.

The parameters observed were fasting blood sugar level and post prandial blood glucose level at baseline, 2 weeks and 4 weeks.

Results 1. Fasting blood sugar (FBS) and post prandial blood glucose levels(PPBS)

The results of the effect of methanol extract of column purified water extract of coconut root on fasting blood sugar and post prandial blood glucose levels were presented in Table 3.

At 2 weeks, the FBS levels in patients treated with methanol extract of column purified water extract of coconut root decreased from 120 mg/dl to 106 mg/dl. Similarly the PPBS level in patients treated with methanol extract of column purified water extract of coconut root decreased from 183 mg/dl to 166 mg/dl. At 4 weeks, the FBS and PPBS decreased to 87 mg/dl and 148 mg/dl respectively.

The results (Table 3) indicated the effect of methanol extract of column purified water extract of coconut root in decreasing fasting blood sugar and post prandial blood glucose levels in patients with impending diabetes mellitus.

Table 3

Example 20( Effect of methanol extract of column purified water extract of coconut root on fasting blood sugar ( FBS) and post prandial blood glucose (PPBS) in diabetic patients)

Newly diagnosed diabetic patients of either sex, aged between 40 and 60 years, and without severe systemic diseases were screened for the study. Among them, 1 8 patients with FBS: 126- 150 mg/dl and PPBS: 200-250 mg/dl, not grossly obese and who gave informed consent were selected and randomized into two groups. A prescribed diet was advised for all the patients throughout the study period Group I - patients treated with glibenclamide 5 mg tablet once daily before breakfast for 4 weeks.

Group II - Patients treated with methanol extract of column purified water extract of coconut root in a 500 mg capsule once daily before breakfast for 4 weeks.

The parameter observed includes fasting blood sugar level and post prandial blood glucose level at baseline, 2 weeks and 4 weeks.

Results

1 . Fasting blood glucose and post prandial blood sugar levels

The results of the effect of methanol extract of column purified water extract of coconut root on fasting blood sugar and post prandial glucose levels on newly diagnosed diabetic patients are presented in Table.4.

At 2 weeks, the FBS levels in diabetic patients treated with methanol extract of column purified water extract of coconut root decreased from 143 mg/dl to 121 mg/dl. Similarly the PPBS level in diabetic patients treated with methanol extract of column purified water extract of coconut root decreased from 227 mg/dl to 184 mg/dl. At 4 weeks, the FBS and PPBS decreased to 86 mg/dl and 158 mg/dl respectively.

The results of the study (Table 4) revealed the effect of methanol extract of column purified water extract of coconut root in decreasing fasting blood glucose and post prandial glucose levels on newly diagnosed diabetic patients at 2 weeks and 4 weeks.

Table 4

We have brought out the novel features of the invention by explaining some of the preferred embodiments under the invention, enabling those in the art to understand and visualize our invention. It is also to be understood that the invention is not limited in its application to the details set forth in the above description or as illustrated in the drawings. Although the invention has been described in considerable detail with reference to certain preferred embodiments thereof, various modifications can be made without departing from the spirit and scope of the invention as described herein above and as defined in the following claims.