Title of Invention	AN IMPROVED MICRO HPLC METHOD FOR IDENTIFICATION OF METFORMIN HYDROCHLORIDE IN ROOT OF CAREYA ARBOREA
Abstract	An improved Micro HPLC method for identifying Metformin hydrochloride in root cover of Careya arborea having the same pharmacological effect for the treatment of Diabetes comprising of a) preparing the standard solution, b) preparing the aqueous solution of root cover of Careya arborea, c) allowed the standard solution of stage (a) and aqueous solution a! stage (b) into Micro HPLC. d) the Micro High Pressure Liquid Chromatography characterized in that column used octadecyl silane, particle sized 3 micrometer, inner diameter of column 5mm, mobile phase of water and methanol (in 50:50 ratio), wavelength 220 nm and flow rate adjusted to 0.2 ml / min. e) the powdered root cover of Careya arborea submerged in distilled water for 6 hours, stirred for 30 minutes followed by filtration and diluted the filtrate twenty times.

Title of Invention

AN IMPROVED MICRO HPLC METHOD FOR IDENTIFICATION OF METFORMIN HYDROCHLORIDE IN ROOT OF CAREYA ARBOREA

Abstract

An improved Micro HPLC method for identifying Metformin hydrochloride in root cover of Careya arborea having the same pharmacological effect for the treatment of Diabetes comprising of a) preparing the standard solution, b) preparing the aqueous solution of root cover of Careya arborea, c) allowed the standard solution of stage (a) and aqueous solution a! stage (b) into Micro HPLC. d) the Micro High Pressure Liquid Chromatography characterized in that column used octadecyl silane, particle sized 3 micrometer, inner diameter of column 5mm, mobile phase of water and methanol (in 50:50 ratio), wavelength 220 nm and flow rate adjusted to 0.2 ml / min. e) the powdered root cover of Careya arborea submerged in distilled water for 6 hours, stirred for 30 minutes followed by filtration and diluted the filtrate twenty times.

Full Text	FORM 2 THE PATENT ACT, 1970 (39 of 1970) [See section 10] (a) Dr. Shipra Roy, A-92, Minal Residency, J.K.Road, Bhopal- 462023 (M.P.) India, Nationality: Indian; (b) Mrs. Venu Agarwal, B-134,Alkapuri, Bhopal - 462024 (M.P.) India, Nationality: Indian TITLE "An improved Micro HPLC method for identification of Metformin hydrochloride in root of Careya arborea." The following specification (particularly) describes the nature of the invention and the manner in which it is to be performed. ORIGINAL 540/MUM/2003 28/05/2003 GRANTED 14-7-2004 Title "An improved Micro HPLC method for identification of Metformin hydrochloride in root of Careya arborea." Field of invention It is a branch of medicinal chemistry, deals with phytochemistry, or plant chemistry, and is concerned with the enormous variety of organic substances that are elaborated and accumulated by plants and deals with the chemical structures of these substances, their biosynthesis, turnover and metabolism, their natural distribution and their biological function. Background of invention It is evident that insulin therapy, as currently practiced, is not a panacea for diabetes mellitus. This realization has promoted a great deal of research toward the development of more effective ways of treating the disease and has led to the discovery of several orally active agents, including the sulphonylureas and the biguanides. The applications of microcolumn HPLC have grown significantly in recent years. The trends in HPLC are to employ continuously smaller diameter columns to reduce solvent consumption, enhance sample detection. The role of microcolumn HPLC is also increasingly important in limited sample. With a very limited sample quantity and sample concentration, micro HPLC offers the best solution for the analysis Object of invention Search for plant drugs, without any side toxic effects. With a very limited sample quantity and sample concentration, capillary HPLC offers the best solution for the analysis. antihyperglycemic agent which improves glucose tolerance in patients with type 2 diabetes, lowering both basal and postprandial plasma glucose. Its pharmacologic mechanisms of action are different from other classes of oral antihyperglycemic agents. Metformin decreases hepatic glucose production, decreases intestinal absorption of glucose, and improves insulin sensitivity by increasing peripheral glucose uptake and utilization. Unlike sulfonylureas, metformin does not produce hypoglycemia in either patients with type 2 diabetes or normal subjects and does not cause hyperinsulinemia. With metformin therapy, insulin secretion remains unchanged while fasting insulin levels and day-long plasma insulin response may actually decrease. Metformin hydrochloride is a biguanide, which does not create any adverse effects on the metabolic reactions.The presence of this metformin hydrochloride was confirmed in the root of Careya arborea was done by using Micro HPLC. Brief Discription Of Accompanying Drawing Drawing 1 shows the peak and the retention time of standard solution of metformin hydrochloride.Drawing 2 shows the similar peak of aqueous solution of Careya arborea. Detailed Description Micro High Pressure Liquid Chromatography was the method employed Large reduction in solvent consumption and therefore solvent operation and toxic waste disposal cost, very low flow rates used with microcolumns make it the method of choice for HPLC-MS, particularly the bench-top MS with electrospray and nanospray interfaces. Low flow rates used in microcolumns facilitate the unified approach to chromatography. These major advantages are the result of the fact that small particles such as 3 and 5 urn can be • efficiently packed in microcolumn and therefore same separation chemistry and elution pattern as that obtained from a 4.6 mm id, column can be expected. As a result of smaller column diameters for microcolumn HPLC, the flow rate requirement is also reduced and the elution peak volume (flow rate times peak width at half-height) The chromatograms obtained during HPLC Studies are represented in drawing 1 and 2. The retention time and characteristic peak was recorded. The standard sample was run in micro HPLC under the similar conditions already mentioned. The test sample was also run under the same experimental conditions. For the test sample at the same retention time the characteristic peak was observed. This confirms the presence of same compound i.e Metformin hydrochloride in the test sample. Thus with the help of micro HPLC , a new method has been evolved with which the presence of a compound in herbal produt having the same pharmacological effect as that of a synthetic compound , can be proved Metformin hydrochloride is a synthetic compound used in synthetic drugs for the treatment of diabetes. This compound was found to be present in the root cover of Careya arborea. The above mentioned method was used for this purpose. Claims We claim (i) An improved Micro HPLC method for identifying Metformin hydrochloride in root cover of Careya arborea having the same pharmacological effect for the treatment of Diabetes comprising of a) preparing the standard solution, b) preparing the aqueous solution of root cover of Careya arborea, c) allowed the standard solution of stage (a) and aqueous solution a! stage (b) into Micro HPLC. d) the Micro High Pressure Liquid Chromatography characterized in that column used octadecyl silane, particle sized 3 micrometer, inner diameter of column 5mm, mobile phase of water and methanol (in 50:50 ratio), wavelength 220 nm and flow rate adjusted to 0.2 ml / min. e) the powdered root cover of Careya arborea submerged in distilled water for 6 hours, stirred for 30 minutes followed by filtration and diluted the filtrate twenty times. (ii) A method as claimed in claim (i) wherein the standard solution prepared using water Dated 26 day of May 2003 Signature (Shipra Roy)

Full Text

FORM 2
THE PATENT ACT, 1970
(39 of 1970)
[See section 10]
(a) Dr. Shipra Roy,
A-92, Minal Residency, J.K.Road, Bhopal- 462023 (M.P.) India,
Nationality: Indian;
(b) Mrs. Venu Agarwal,
B-134,Alkapuri, Bhopal - 462024 (M.P.) India, Nationality: Indian
TITLE
"An improved Micro HPLC method for identification of Metformin hydrochloride in root of Careya arborea."
The following specification (particularly) describes the nature of the invention and the manner in which it is to be performed.

ORIGINAL
540/MUM/2003
28/05/2003
GRANTED
14-7-2004

Title
"An improved Micro HPLC method for identification of Metformin hydrochloride in root of Careya arborea."
Field of invention
It is a branch of medicinal chemistry, deals with phytochemistry, or plant
chemistry, and is concerned with the enormous variety of organic substances that are elaborated and accumulated by plants and deals with the chemical structures of these substances, their biosynthesis, turnover and metabolism, their natural distribution and their biological function.
Background of invention
It is evident that insulin therapy, as currently practiced, is not a panacea
for diabetes mellitus. This realization has promoted a great deal of research toward the development of more effective ways of treating the disease and has led to the discovery of several orally active agents, including the sulphonylureas and the biguanides.
The applications of microcolumn HPLC have grown significantly in recent years. The trends in HPLC are to employ continuously smaller diameter columns to reduce solvent consumption, enhance sample detection. The role of microcolumn HPLC is also increasingly important in limited sample. With a very limited sample quantity and sample concentration, micro HPLC offers the best solution for the analysis
Object of invention
Search for plant drugs, without any side toxic effects. With a very limited
sample quantity and sample concentration, capillary HPLC offers the best solution for the analysis.

antihyperglycemic agent which improves glucose tolerance in patients with type 2 diabetes, lowering both basal and postprandial plasma glucose. Its pharmacologic mechanisms of action are different from other classes of oral antihyperglycemic agents. Metformin decreases hepatic glucose production, decreases intestinal absorption of glucose, and improves insulin sensitivity by increasing peripheral glucose uptake and utilization. Unlike sulfonylureas, metformin does not produce hypoglycemia in either patients with type 2 diabetes or normal subjects and does not cause hyperinsulinemia. With metformin therapy, insulin secretion remains unchanged while fasting insulin levels and day-long plasma insulin response may actually decrease. Metformin hydrochloride is a biguanide, which does not create any adverse effects on the metabolic reactions.The presence of this metformin hydrochloride was confirmed in the root of Careya arborea was done by using Micro HPLC.
Brief Discription Of Accompanying Drawing
Drawing 1 shows the peak and the retention time of standard solution of metformin hydrochloride.Drawing 2 shows the similar peak of aqueous solution of Careya arborea.
Detailed Description
Micro High Pressure Liquid Chromatography was the method employed Large reduction in solvent consumption and therefore solvent operation and toxic waste disposal cost, very low flow rates used with microcolumns make it the method of choice for HPLC-MS, particularly the bench-top MS with electrospray and nanospray interfaces. Low flow rates used in microcolumns facilitate the unified approach to chromatography. These major advantages are the result of the fact that small particles such as 3 and 5 urn can be • efficiently packed in microcolumn and therefore same separation chemistry and elution pattern as that obtained from a 4.6 mm id, column can be expected. As a result of smaller column diameters for microcolumn HPLC, the flow rate requirement is also reduced and the elution peak volume (flow rate times peak width at half-height)

The chromatograms obtained during HPLC Studies are represented in drawing 1 and 2. The retention time and characteristic peak was recorded. The standard sample was run in micro HPLC under the similar conditions already mentioned. The test sample was also run under the same experimental conditions. For the test sample at the same retention time the characteristic peak was observed. This confirms the presence of same compound i.e Metformin hydrochloride in the test sample.
Thus with the help of micro HPLC , a new method has been evolved with which the presence of a compound in herbal produt having the same pharmacological effect as that of a synthetic compound , can be proved
Metformin hydrochloride is a synthetic compound used in synthetic drugs for the treatment of diabetes. This compound was found to be present in the root cover of Careya arborea. The above mentioned method was used for this purpose.

Claims
We claim
(i) An improved Micro HPLC method for identifying Metformin hydrochloride in root cover of Careya arborea having the same pharmacological effect for the treatment of Diabetes comprising of
a) preparing the standard solution,
b) preparing the aqueous solution of root cover of Careya arborea,

c) allowed the standard solution of stage (a) and aqueous solution a! stage (b) into Micro HPLC.
d) the Micro High Pressure Liquid Chromatography characterized in that column used octadecyl silane, particle sized 3 micrometer, inner diameter of column 5mm, mobile phase of water and methanol (in 50:50 ratio), wavelength 220 nm and flow rate adjusted to 0.2 ml / min.
e) the powdered root cover of Careya arborea submerged in distilled water for 6 hours, stirred for 30 minutes followed by filtration and diluted the filtrate twenty times.
(ii) A method as claimed in claim (i) wherein the standard solution
prepared using water Dated 26 day of May 2003

Signature
(Shipra Roy)

Documents:

540-mum-2003-cancelled pages(14-7-2004).pdf

540-mum-2003-claims(granted)-(14-7-2004).doc

540-mum-2003-claims(granted)-(14-7-2004).pdf

540-mum-2003-correspondence(14-7-2004).pdf

540-mum-2003-correspondence(ipo)-(4-5-2007).pdf

540-mum-2003-form 1(6-4-2004).pdf

540-mum-2003-form 19(28-5-2003).pdf

540-mum-2003-form 2(granted)-(14-7-2004).doc

540-mum-2003-form 2(granted)-(14-7-2004).pdf

540-mum-2003-form 3(23-6-2003).pdf

540-mum-2003-form 3(26-5-2003).pdf

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Patent Number

206666

Indian Patent Application Number

540/MUM/2003

PG Journal Number

42/2008

Publication Date

17-Oct-2008

Grant Date

04-May-2007

Date of Filing

28-May-2003

Name of Patentee

DR. SHIPRA ROY

Applicant Address

A-92, MINAL RESIDENCY, J.K. ROAD, BHOPAL,

Inventors:

#	Inventor's Name	Inventor's Address
1	DR. SHIPRA ROY	A-92, MINAL RESIDENCY, J.K. ROAD, BHOPAL - 462023
2	MRS. VENU AGARWAL	B-134, ALKAPURI, BHOPAL - 462024,

PCT International Classification Number

A61B 3/00

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA