Title of Invention	A PROCESS FOR EXTRACTION OF BIOACTIVE COMPOUNDS-POLYPHENOLS, POLYPHENOL OXIDASE, PEROXIDASE, DIETARY FIBRE FROM MANGO PEEL
Abstract	The present invention relates to a process for the extraction of bioactive compounds - polyphenols, polyphenol oxidase, peroxidase, dietary fibre from mango peel. This process finds application in mango processing industry and food and nutrition industry. The process steps are: homogenizing the mango peel in a buffer pH 7.5,treating homogenate with polar organic solvent, filtering resulting slurry to yield mango peel powder and organic solvent containing polyphenols, air drying the resulting powder and extraction of the mango peel powder with buffer, separating the extract into supernatant and residue by centrifugation, recovering the bioactive compound- polyphenols, polyphenol oxidase, peroxidase, dietary fibre by conventional methods.

Title of Invention

A PROCESS FOR EXTRACTION OF BIOACTIVE COMPOUNDS-POLYPHENOLS, POLYPHENOL OXIDASE, PEROXIDASE, DIETARY FIBRE FROM MANGO PEEL

Abstract

The present invention relates to a process for the extraction of bioactive compounds - polyphenols, polyphenol oxidase, peroxidase, dietary fibre from mango peel. This process finds application in mango processing industry and food and nutrition industry. The process steps are: homogenizing the mango peel in a buffer pH 7.5,treating homogenate with polar organic solvent, filtering resulting slurry to yield mango peel powder and organic solvent containing polyphenols, air drying the resulting powder and extraction of the mango peel powder with buffer, separating the extract into supernatant and residue by centrifugation, recovering the bioactive compound- polyphenols, polyphenol oxidase, peroxidase, dietary fibre by conventional methods.

Full Text	The present invention relates to a process for extraction of bioactive compounds-polyphenols, polyphenol oxidase, peroxidase, dietary fibre from mango peel. This process finds application in mango processing industry and food and nutrition industry. Mango is a very important tropical fruit and India is the major producer in the world. It is used in various food preparations such as pulp, amchur and pickles. During processing of mango fruits for pulp, peel is a by-product which contributes 20-25 % of the whole fruit. As it is currently not being utilized for any commercial purpose it is discarded and becoming a source of pollution. Therefore, disposal has become expensive because of transportation. R. Pedroza, Isla, E. Arguillar-Esperanaza, E.J. Vernon-Carter, AICHE Symposium Series, 1994, Vol.90 (300), pages 36-41, have utilized peel for the production of pectin. In an another article, N.R.Raju, S. Sumitra Devi, Krishanand, Biotechnolgy Letters, 1991, Vol. 13, pages 461-464, have used peel for the biogas production. However, mango peel is a good source of nutrients. Reference may be made to the article by Tandon et al., Indian Food Packer, 1991, Vol.45, pages 9-12, which states that mango peel contains good quality pectin. Recently, in another article Laurrauri et al., Z. Lebensmittel Unters Forsch A, 1997, Vol. 205, pages 39-42, have reported that mango peel dietary fibre has been associated with certain natural anti-oxidants and its anti-oxidant capacity was higher than that of α-tocopherol. Laurrauri, 2nd International Electronic Conference on Synthetic organic chemistry (ECSOC-2), Sept 1-30, Arizona, U.S.A., 1988, has reported that mango peel is also a good source of polyphenols. In another article, T.N. Prabha and Patwardhan, J.Biosciences, vol.4, 1982, pages 69-78, have reported the presence of polyphenol oxidase activity in mango peel. Recently, in another article Saby John et al., J Food Biochemistry, 2002, Vol. 26, pages 403-414, have also reported that mango peel contains polyphenol oxidase, peroxidase and polyphenols. Thus, it is evident that mango peel, a by-product of mango industry, could be a valuable source of number of bioactive compounds. The present innovation pertains to the development of a simple process to isolate value added products viz., polyphenols, polyphenol oxidase, peroxidase, dietary fibre simultaneously from the mango peel. The unit of activity 'U' is well known in prior art. Accordingly, a process for extraction of bioactive compounds- polyphenols, polyphenol oxidase, peroxidase, dietary fibre from mango peel comprises: a) homogenizing the mango peel in a buffer pH 7.5, b) treatment of homogenate in a ratio of 1:4 (w/v) with polar organic solvent, c) filtering resulting slurry to yield mango peel powder and organic solvent containing polyphenols d) air drying of the resulting powder and extraction of the mango peel powder with buffer; and e) separating the extract into supernatant and residue by centrifugation, and the supernatant obtained contains protein/enzymes and the residue consists mainly of dietary fibre. f) recovering the bioactive compound by conventional methods In an embodiment of the process the organic solvent used may be acetone or ethyl alcohol, In an another embodiment of the process wherein the extraction of soluble portion from the peel powder may be done in 50 mM sodium phosphate buffer, In an another embodiment of the process, wherein the organic solvent extract may contain polyphenols ranging from 11 mg to 22 mg /g peel, In an another embodiment of the process, wherein aqueous extract of organic solvent powder may contain peroxidase ranging from 56 U to 388 U/g mango peel, In an another embodiment of the process, wherein aqueous extract of organic solvent powder may contain polyphenol oxidase ranging from 34 U to 106U/g mango peel, In an another embodiment of the process, wherein the soluble dietary fibre range from 11 %to23%. Following examples are given by the way of illustration of the present invention and therefore, should not be construed to limit for the scope of the present invention. Example 1 Mango peel of the 'Raspuri' mango variety was collected. Mango peels were cut into smaller pieces. Cut mango peel was homogenized in a blender (100g peel/50 ml) with sodium phosphate (50 mM, pH 7.5) solution. The homogenate thus obtained was treated with 4 volumes of acetone that was chilled to -10 °C to -15°C. The resultant slurry was filtered through a Buckner funnel. The residue obtained was washed once again with 300 ml of chilled acetone. The residue obtained was air-dried. The acetone extract was stored at 4°C. Phenolic content in organic solvent was estimated. Dry powder (1g) was extracted with 50 ml of sodium phosphate buffer (pH 7.5; 50 mM). Insoluble components were separated by the centrifugation. The soluble fractions was analysed for protein and enzymes viz., peroxidase and polyphenol oxidase activity. The insoluble residue was analysed for dietary fibre content. Example No. 2 Mango peel of the 'Malgoa' mango variety was collected. Mango peels were cut into smaller pieces. Cut mango peel was homogenized in a blender (100g peel/50 ml) with sodium phosphate (50 mM, pH 7.5) solution. The homogenate thus obtained was treated with 4 volumes of acetone that was chilled to -10 °C to -15 °C. The resultant slurry was filtered through a Buckner funnel. The residue obtained was washed once again with 300 ml of chilled acetone. The residue obtained was airdried. The acetone extract was stored at 4°C. Phenolic content in acetone was estimated. Dry powder (1g) was extracted with 50 ml of sodium phosphate buffer (pH 7.5; 50 mM). Insoluble components were separated by the centrifugation. The soluble fractions were analyzed for protein and enzymes viz., peroxidase (POD) and polyphenol oxidase (PPO) activity. The insoluble residue was analyzed for dietary fibre content. Example No. 3 Mango peel of the 'Badami' mango variety was collected. Mango peels were cut into smaller pieces and made into two portions. One portion was homogenized in a blender (100g peel/50 ml) with sodium phosphate (50 mM, pH 7.5) solution and the homogenate thus obtained was treated with 4 volumes of acetone that was chilled to -10°C to -15°C. The resultant slurry was filtered through a Buckner funnel. The residue obtained was washed once again with 300 ml of chilled acetone. The residue obtained was air-dried. The acetone extract was stored at 4°C. Another portion of peel was homogenized in a blender with sodium phosphate (50 mM, pH 7.5) solution (100g peel/50 ml) and the homogenate thus obtained was treated with 4 volumes of ethyl alcohol that was chilled to -10 °C to -15 °C. The resultant slurry was filtered through a Buckner funnel. The residue obtained was washed once again with 300 ml of chilled ethyl alcohol. The residue obtained was air-dried. The ethyl alcohol extract was stored at 4°C. Phenolic content in acetone and ethyl alcohol extracts were estimated separately. Dry powder (1g) of each was extracted with 100 ml of sodium phosphate buffer (pH 7.5; 50 mM). Insoluble components were separated by the centrifugation. The soluble fractions were analyzed for protein and enzymes viz., peroxidase (POD) and polyphenol oxidase (PPO) activity. The insoluble residue was analyzed for dietary fibre content. Advantages (i) In mango industry especially pulping and amchur powder preparation, mango peel is a waste by-product and creating environmental pollution. The process developed uses this waste material and produces valuable bioactive / food ingredients, (ii) The different value added products isolated from the same starting material should make the process more economical. We claim: 1) A process for extraction of bioactive compounds viz.- polyphenols, polyphenol oxidase, peroxidase, dietary fibre from the mango peels which comprises: a) homogenizing the mango peel in a buffer pH 7.5, b) treatment of homogenate in a ratio of 1:4 (w/v) with polar organic solvent, c) filtering resulting slurry to yield mango peel powder and organic solvent containing polyphenols, d) air drying of the resulting powder and extraction of the mango peel powder with buffer, e) separating the extract into supernatant and residue by centrifugation, and the supernatant obtained contains protein/enzymes and the residue consists mainly of dietary fibre, f) recovering the bioactive compound- polyphenols, polyphenol oxidase, peroxidase, dietary fibre by conventional methods, 2) A process as claimed in claim 1, wherein organic solvent used is acetone or ethyl alcohol, 3) A process as claimed in claim 1, wherein the extraction of soluble portion from the peel powder is done in 50 mM sodium phosphate buffer, 4) A process as claimed in claim 1, wherein the organic solvent extract contains polyphenols ranging from 11 mg to 22 mg /g peel. 5) A process as claimed in claim 1, wherein aqueous extract of organic solvent powder contains peroxidase ranging from 56 U to 388 U/g mango peel, 6) A process as claimed in claim 1, wherein aqueous extract of organic solvent powder contains polyphenol oxidase ranging from 34 U to 106U/g mango peel, 7) A process as claimed in claims 1 or 8, wherein the soluble dietary fibre range from 11 % to 23 %, 8) A process for extraction of bioactive compounds viz.- polyphenols, polyphenol oxidase, peroxidase, dietary fibre from the mango peels substantially as herein described with reference to the examples.

Full Text

The present invention relates to a process for extraction of bioactive compounds-polyphenols, polyphenol oxidase, peroxidase, dietary fibre from mango peel. This process finds application in mango processing industry and food and nutrition industry.
Mango is a very important tropical fruit and India is the major producer in the world. It is used in various food preparations such as pulp, amchur and pickles. During processing of mango fruits for pulp, peel is a by-product which contributes 20-25 % of the whole fruit. As it is currently not being utilized for any commercial purpose it is discarded and becoming a source of pollution. Therefore, disposal has become expensive because of transportation. R. Pedroza, Isla, E. Arguillar-Esperanaza, E.J. Vernon-Carter, AICHE Symposium Series, 1994, Vol.90 (300), pages 36-41, have utilized peel for the production of pectin. In an another article, N.R.Raju, S. Sumitra Devi, Krishanand, Biotechnolgy Letters, 1991, Vol. 13, pages 461-464, have used peel for the biogas production.
However, mango peel is a good source of nutrients. Reference may be made to the article by Tandon et al., Indian Food Packer, 1991, Vol.45, pages 9-12, which states that mango peel contains good quality pectin. Recently, in another article Laurrauri et al., Z. Lebensmittel Unters Forsch A, 1997, Vol. 205, pages 39-42, have reported that mango peel dietary fibre has been associated with certain natural anti-oxidants and its anti-oxidant capacity was higher than that of α-tocopherol. Laurrauri, 2nd International Electronic Conference on Synthetic organic chemistry (ECSOC-2), Sept 1-30, Arizona, U.S.A., 1988, has reported that mango peel is also a good source of polyphenols. In another article, T.N. Prabha and Patwardhan, J.Biosciences, vol.4, 1982, pages 69-78, have

reported the presence of polyphenol oxidase activity in mango peel. Recently, in another article Saby John et al., J Food Biochemistry, 2002, Vol. 26, pages 403-414, have also reported that mango peel contains polyphenol oxidase, peroxidase and polyphenols. Thus, it is evident that mango peel, a by-product of mango industry, could be a valuable source of number of bioactive compounds. The present innovation pertains to the development of a simple process to isolate value added products viz., polyphenols, polyphenol oxidase, peroxidase, dietary fibre simultaneously from the mango peel. The unit of activity 'U' is well known in prior art. Accordingly, a process for extraction of bioactive compounds- polyphenols, polyphenol oxidase, peroxidase, dietary fibre from mango peel comprises:
a) homogenizing the mango peel in a buffer pH 7.5,
b) treatment of homogenate in a ratio of 1:4 (w/v) with polar organic solvent,
c) filtering resulting slurry to yield mango peel powder and organic solvent containing polyphenols
d) air drying of the resulting powder and extraction of the mango peel powder with buffer; and
e) separating the extract into supernatant and residue by centrifugation, and the supernatant obtained contains protein/enzymes and the residue consists mainly of dietary fibre.
f) recovering the bioactive compound by conventional methods

In an embodiment of the process the organic solvent used may be acetone or
ethyl alcohol,
In an another embodiment of the process wherein the extraction of soluble
portion from the peel powder may be done in 50 mM sodium phosphate buffer,
In an another embodiment of the process, wherein the organic solvent extract
may contain polyphenols ranging from 11 mg to 22 mg /g peel,
In an another embodiment of the process, wherein aqueous extract of organic
solvent powder may contain peroxidase ranging from 56 U to 388 U/g mango
peel,
In an another embodiment of the process, wherein aqueous extract of organic
solvent powder may contain polyphenol oxidase ranging from 34 U to 106U/g
mango peel,
In an another embodiment of the process, wherein the soluble dietary fibre range
from 11 %to23%.
Following examples are given by the way of illustration of the present invention
and therefore, should not be construed to limit for the scope of the present
invention.
Example 1
Mango peel of the 'Raspuri' mango variety was collected. Mango peels were cut
into smaller pieces. Cut mango peel was homogenized in a blender (100g
peel/50 ml) with sodium phosphate (50 mM, pH 7.5) solution. The homogenate
thus obtained was treated with 4 volumes of acetone that was chilled to -10 °C
to -15°C. The resultant slurry was filtered through a Buckner funnel. The residue
obtained was washed once again with 300 ml of chilled acetone. The residue
obtained was air-dried. The acetone extract was stored at 4°C.
Phenolic content in organic solvent was estimated. Dry powder (1g) was
extracted with 50 ml of sodium phosphate buffer (pH 7.5; 50 mM). Insoluble
components were separated by the centrifugation. The soluble fractions was
analysed for protein and enzymes viz., peroxidase and polyphenol oxidase
activity. The insoluble residue was analysed for dietary fibre content.
Example No. 2
Mango peel of the 'Malgoa' mango variety was collected. Mango peels were cut into
smaller pieces. Cut mango peel was homogenized in a blender (100g peel/50 ml)
with sodium phosphate (50 mM, pH 7.5) solution. The homogenate thus obtained
was treated with 4 volumes of acetone that was chilled to -10 °C to -15 °C. The
resultant slurry was filtered through a Buckner funnel. The residue obtained was
washed once again with 300 ml of chilled acetone. The residue obtained was airdried.
The acetone extract was stored at 4°C.
Phenolic content in acetone was estimated. Dry powder (1g) was extracted with 50
ml of sodium phosphate buffer (pH 7.5; 50 mM). Insoluble components were
separated by the centrifugation. The soluble fractions were analyzed for protein and
enzymes viz., peroxidase (POD) and polyphenol oxidase (PPO) activity. The
insoluble residue was analyzed for dietary fibre content.
Example No. 3
Mango peel of the 'Badami' mango variety was collected. Mango peels were cut into
smaller pieces and made into two portions. One portion was homogenized in a
blender (100g peel/50 ml) with sodium phosphate (50 mM, pH 7.5) solution and the
homogenate thus obtained was treated with 4 volumes of acetone that was chilled to
-10°C to -15°C. The resultant slurry was filtered through a Buckner funnel. The
residue obtained was washed once again with 300 ml of chilled acetone. The residue
obtained was air-dried. The acetone extract was stored at 4°C.
Another portion of peel was homogenized in a blender with sodium phosphate (50
mM, pH 7.5) solution (100g peel/50 ml) and the homogenate thus obtained was
treated with 4 volumes of ethyl alcohol that was chilled to -10 °C to -15 °C. The
resultant slurry was filtered through a Buckner funnel. The residue obtained was
washed once again with 300 ml of chilled ethyl alcohol. The residue obtained was
air-dried. The ethyl alcohol extract was stored at 4°C.
Phenolic content in acetone and ethyl alcohol extracts were estimated separately.
Dry powder (1g) of each was extracted with 100 ml of sodium phosphate buffer (pH
7.5; 50 mM). Insoluble components were separated by the centrifugation. The
soluble fractions were analyzed for protein and enzymes viz., peroxidase (POD) and
polyphenol oxidase (PPO) activity. The insoluble residue was analyzed for dietary
fibre content.
Advantages
(i) In mango industry especially pulping and amchur powder preparation, mango peel
is a waste by-product and creating environmental pollution. The process developed
uses this waste material and produces valuable bioactive / food ingredients,
(ii) The different value added products isolated from the same starting material
should make the process more economical.

We claim:
1) A process for extraction of bioactive compounds viz.- polyphenols,
polyphenol oxidase, peroxidase, dietary fibre from the mango peels which
comprises:
a) homogenizing the mango peel in a buffer pH 7.5,
b) treatment of homogenate in a ratio of 1:4 (w/v) with polar organic solvent,

c) filtering resulting slurry to yield mango peel powder and organic solvent containing polyphenols,
d) air drying of the resulting powder and extraction of the mango peel powder with buffer,
e) separating the extract into supernatant and residue by centrifugation, and the supernatant obtained contains protein/enzymes and the residue consists mainly of dietary fibre,
f) recovering the bioactive compound- polyphenols, polyphenol oxidase, peroxidase, dietary fibre by conventional methods,

2) A process as claimed in claim 1, wherein organic solvent used is acetone or ethyl alcohol,
3) A process as claimed in claim 1, wherein the extraction of soluble portion from the peel powder is done in 50 mM sodium phosphate buffer,
4) A process as claimed in claim 1, wherein the organic solvent extract contains polyphenols ranging from 11 mg to 22 mg /g peel.

5) A process as claimed in claim 1, wherein aqueous extract of organic solvent powder contains peroxidase ranging from 56 U to 388 U/g mango peel,
6) A process as claimed in claim 1, wherein aqueous extract of organic solvent powder contains polyphenol oxidase ranging from 34 U to 106U/g mango peel,
7) A process as claimed in claims 1 or 8, wherein the soluble dietary fibre range from 11 % to 23 %,
8) A process for extraction of bioactive compounds viz.- polyphenols, polyphenol oxidase, peroxidase, dietary fibre from the mango peels substantially as herein described with reference to the examples.

Documents:

443-DEL-2003-Abstract-(17-10-2008).pdf

443-del-2003-abstract.pdf

443-DEL-2003-Claims-(17-10-2008).pdf

443-del-2003-claims.pdf

443-DEL-2003-Correspondence-Others-(17-10-2008).pdf

443-del-2003-correspondence-others.pdf

443-del-2003-correspondence-po.pdf

443-DEL-2003-Description (Complete)-(17-10-2008).pdf

443-del-2003-description (complete).pdf

443-DEL-2003-Form-1-(17-10-2008).pdf

443-del-2003-form-1.pdf

443-del-2003-form-18.pdf

443-DEL-2003-Form-2-(17-10-2008).pdf

443-del-2003-form-2.pdf

443-del-2003-form-3.pdf

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Patent Number

225497

Indian Patent Application Number

443/DEL/2003

PG Journal Number

48/2008

Publication Date

28-Nov-2008

Grant Date

14-Nov-2008

Date of Filing

26-Mar-2003

Name of Patentee

COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH

Applicant Address

RAFI MARG, NEW DELHI-110001, INDIA.

Inventors:

#	Inventor's Name	Inventor's Address
1	UMMITI JAYASANKARA PRASADA RAP	DELHI, INDIA.
2	SANTHOOR GURURAJA BHAT	DELHI, INDIA.
3	AJILA CHANDRAN MATHEYAM BATH	DELHI, INDIA.
4	MAHADEVAMMA	DELHI, INDIA.

PCT International Classification Number

A61K 35/78

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA