Title of Invention	"A PROCESS FOR THE PREPARATION OF ANTIOXIDANTS FROM GARCINIA"
Abstract	The invention relates to a process for the preparation of xanthon antioxidants from Garcinia. This is for the first time that antioxidants has been extracted from Garcinia cowa. The process steps are: extracting the rinds of Garcinia cowa with non polar solvent in a Soxhlet extractor at temperature in the range of 55-60°C for a period of 4-8 hr, filtering the above extract to obtain the particle free extract, concentrating the above particle free extract at a temperature of 35-40°C under vacuum may be effected to recover the solvent to an extent of 95%, drying the above concentrated extracted using vacuum oven at 40-45°C under vacuum to obtain xanthon antioxidant.

Title of Invention

"A PROCESS FOR THE PREPARATION OF ANTIOXIDANTS FROM GARCINIA"

Abstract

The invention relates to a process for the preparation of xanthon antioxidants from Garcinia. This is for the first time that antioxidants has been extracted from Garcinia cowa. The process steps are: extracting the rinds of Garcinia cowa with non polar solvent in a Soxhlet extractor at temperature in the range of 55-60°C for a period of 4-8 hr, filtering the above extract to obtain the particle free extract, concentrating the above particle free extract at a temperature of 35-40°C under vacuum may be effected to recover the solvent to an extent of 95%, drying the above concentrated extracted using vacuum oven at 40-45°C under vacuum to obtain xanthon antioxidant.

Full Text	The present invention relates to a process for the preparation of antioxidants from Garcinia. Overproduction of oxidants in certain conditions can cause an imbalance, leading to oxidative damage to large biomolecules such as lipids, DNA, and proteins. The consequences of such extensive damages are involved in the development of various degenerative disease states like atherosclerosis, cardiovascular disease, cataracts, tissue damage in rheumatoid arthritis, decline in immune function, dysfunction of brain, and certain types of cancer. Besides the endogenous defenses, consumption of dietary antioxidants could be an important aspect of body's defense mechanism to protect against reactive oxygen species (ROS) that produced under normal metabolism and pathophysiological conditions and also many antioxidants are being identified as anticarcinogens. Diets rich in fruits and vegetables have been associated with lower incidence and mortality rates of cancer and low dietary intake of the same doubles the risk of cancer as compared to high intake (Ames, B.N., Shigenaga, M.K., & Hagen, T.M. Oxidants antioxidants and degenerative diseases of aging. Proc. Natl. Acad. Sci. USA, 90, 7915-7922, 1993). It is generally assumed that the active dietary constituents contributing to the protective effects are the antioxidant nutrients. It is important to note that most of the investigations regarding inhibitory effect of food component on the oxidative damages of biomolecules have been devoted to the foods of plant origin. Oxidation reactions are of major significance in human physiology as well as the food industry. In the former, oxidative stress has been linked to diseases while food industry long been concerned with issues such as rancidity and oxidative spoilage of fruits and vegetables, beverages and other food stuffs (Shahidi, P.; Wanasundara, P.K. (1992). Grit. Rev. Food Sc. Nutr, 32, 67-103). The widely used food antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), tertbutylhydroxyquinone (TBHQ) and alkyl gallates are synthetic compounds of phenolic nature. Synthetic antioxidants, like BHA and BHT, have restricted use in foods as these synthetic antioxidants are suspected to be carcinogenic (Madhavi, D. L; Salunkhe, D. K. Toxicological Aspects of Food Antioxidants, In-Foocf Antioxidants, Eds.; Madhavi, D. L., Deshpande, S. S. Salunkhe, D. K., Marcel Dekker Inc, New York, 1995, p.267). Consumer concerns and government regulations are fostering the research on alternative ingredients, mostly natural polyphenols, as presumably safer food antioxidants with added beneficial functional properties. Thus the discovery of new natural antioxidants has obvious nutritional and pharmaceutical relevance. Garcinia (Family: Guff/ferae) is a large genus of polygamous trees or shrubs, distributed in the tropical Asia, Africa and Polynesia. It consists of 180 species, out of which about 30 species are found in India (The Wealth of India -Raw Materials, Vol.IV, CSIR, New Delhi, India, 1956, pp.99-108). Among the species Garcinia cowa is found in Northeastern part of India and Andaman Islands. In upper Assam, it is often cultivated in homesteads for its acid fruit. The fruits are edible, dried slices of the fruit are used in curry in place of tamarind/lemon. Garcinia is a rich source of secondary metabolites including xanthones, flavonoids, benzophenones, lactones and phenolic acids. Since most xanthones have phenolic functional groups on a linear tricyclic ring they often exhibit a wide range of biological and pharmacological activities (Bennet, G.J.; Lee, H.H. Xanthones from Guttiferae. Phytochemistry, 28, 967-998, 1989; Minami, H., Kinoshita, M., Fukuyama, Y., Kodama, M., Yoshizawa, T., Sugiura, M. Nakagawa, K. and Tago, H. 1994, Antioxidant xanthones from Garcinia subelliptica. Phytochemistry 36, 501-506). In the background of synthetic antioxidants being used in foods, there is a need to explore the possibility of isolation of antioxidants from plant sources. To overcome this drawback, the present invention provides a process for preparation of antioxidants from Garcinia, wherein the process relates to the preparation of antioxidants from the rinds of Garcinia cowa. The main object of the present invention is to provide a process for the preparation of antioxidants from Garcinia, wherein rinds Garcinia cowa are used in the preparation of natural antioxidants. An object of the present invention is to use simple extraction methods and solvents used for the preparation of antioxidants. Another object of the present invention is to produce the antioxidants, which is essentially neutral in flavour and dark brown in colour. The process is efficient and itinvolves the use of simple extraction methods and solvents, which can be regenerated. Accordingly, the present invention provides a process for the preparation of xanthon antioxidants from Garcinia, which comprises : i) selecting the rinds of Garcinia cowa ii) characterized in that cutting the rinds of Garcinia cowa manually to a size of 4x6 mm to 8x10 mm, iii) extracting of the above said material (ii) with non polar solvent in a Soxhlet extractor at temperature in the range of 55-60°C for a period of 4-8 hr, iv) filtering the above extract to obtain the particle free extract, v) concentrating the above particle free extract at a temperature of 35-40°C under vacuum may be effected to recover the solvent to an extent of 95%, vi) drying the above concentrated extracted using vacuum oven at 40-45°C under vacuum at 8-27 mm of mercury to obtain xanthon antioxidant. In an embodiment of the present invention, the yield of chloroform extract may be in the range of 5.5-8.0%. In an embodiment of the present invention, the product has antioxidant activity as potent as or more potent than butylated hydroxyanisole (BHA) is useful in food and medicine. The preparation of antioxidants from Garcinia cowa was done according to following flow diagram: Rinds of Garcinia cowa Soxhlet extractor Non-polar solvents at 55-60 °C Extract Concentrated at 35-40°C and 10-25 mm of mercury Spent It can be used for Hydroxycitric acid extraction Dried under vacuum at 40-50 °C and 10-25 mm of mercury Yield of antioxidant fraction (6.0-7.5%) Recovery of solvent The novelty of the process is 1. This is the first report of preparation of antioxidants from G. cowa. 2. The invention is a single step process to obtain the antioxidant fraction from G. cowa. The advantages of the process is 1. Preparation of natural antioxidants 2. The process is simple and the solvents used in this process can be regenerated for further use. 3. The left over spent can be further used for the extraction of (-)-Hydroxycitric acid. The following examples are given by way of illustration of the present invention and therefore should not be constructed to limit the scope of the present invention. Example -1 Fifty grams of rinds of Garcinia cowa was manually cut into small pieces and extracted using 200 ml of cyclohexane at 60 °C for 4 h in a Soxhlet extractor. The extract was filtered using Whatman filter paper No.1. The extract was concentrated at a temperature of 40 °C and under redyced pressure at 10 mm of mercury to recover the solvent and dried in vacuum oven at 40 °C and 25 mm of mercury. The yield of antioxidant extract was 3.1 g. Example -2 The dried rinds in 100 g aliquots of Garcinia cowa were extracted with 400 ml of hexane by using Soxhlet extractor at 55 °C for 8 h. The extract was collected and filtered through Whatman filter paper no. 1. The filtrate was concentrated at a temperature of 35 °C and under a reduced pressure at 25 mm of mercury to recover the solvent and dried in vacuum oven at a temperature of 50 °C and under vacuum of 10 mm of mercury. The yield of extract was 6.4 g. The extract obtained from the above method was screened for antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) (Singh et al., J. Agricultural and Food Chemistry, 50, 81-86, 2002). At 10 ppm concentration, both BHA and the extract of G. cowa showed 76 and 75.7% respectively, free radical scavenging activity. Example -3 Rinds in quantities of 150 g of Garcinia cowa were extracted with 600 ml of benzene by using Soxhlet extractor at 60 °C for 8 h. The extract was filtered through Whatman filter paper no. 1. The filtrate was concentrated at a temperature of 40 °C and under reduced pressure at 10 mm of mercury to recover the solvent and dried in a vacuum oven at 40 °C under vacuum at 25 mm of mercury. The yield of benzene extract was 8.2 g. The benzene extract obtained from the above method was screened for antioxidant activity using (3-carotene-linoleate model systems. BHA and benzene extract showed 98 and 93.7% antioxidant activity at 200 ppm using (3-carotenelinoleate method (Jayaprakasha G.K. and Jaganmohan Rao L. Phenolic constituents from lichen Parmotrema stuppeum (Nyl.) Hale and their antioxidant activity, Zeitschrift fur Naturforschung, 55c, 1018-1022, 2000). We claim : 1. A process for the preparation of xanthon antioxidants from Garcinia, which comprises : i) selecting the rinds of Garcinia cowa ii) characterized in that cutting the rinds of Garcinia cowa manually to a size of 4x6 mm to 8x10 mm, iii) extracting of the above said material (ii) with non polar solvent in a Soxhlet extractor at temperature in the range of 55-60°C for a period of 4-8 hr, iv) filtering the above extract to obtain the particle free extract, v) concentrating the above particle free extract at a temperature of 35-40°C under vacuum may be effected to recover the solvent to an extent of 95%, vi) drying the above concentrated extract using vacuum oven at 40- 45°C under vacuum at 8-27 mm of mercury to obtain xanthon antioxidant. 2. A process as claimed in claim 1, wherein the non polar solvent used is hexane, cyclohexane, pentane, benzene. 3. A process for the preparation of xanthon antioxidants from Garcinia substantially as herein described with reference to the examples.

Full Text

The present invention relates to a process for the preparation of antioxidants
from Garcinia.
Overproduction of oxidants in certain conditions can cause an imbalance,
leading to oxidative damage to large biomolecules such as lipids, DNA, and proteins.
The consequences of such extensive damages are involved in the development of
various degenerative disease states like atherosclerosis, cardiovascular disease,
cataracts, tissue damage in rheumatoid arthritis, decline in immune function,
dysfunction of brain, and certain types of cancer. Besides the endogenous defenses,
consumption of dietary antioxidants could be an important aspect of body's defense
mechanism to protect against reactive oxygen species (ROS) that produced under
normal metabolism and pathophysiological conditions and also many antioxidants
are being identified as anticarcinogens. Diets rich in fruits and vegetables have been
associated with lower incidence and mortality rates of cancer and low dietary intake
of the same doubles the risk of cancer as compared to high intake (Ames, B.N.,
Shigenaga, M.K., & Hagen, T.M. Oxidants antioxidants and degenerative diseases of
aging. Proc. Natl. Acad. Sci. USA, 90, 7915-7922, 1993). It is generally assumed
that the active dietary constituents contributing to the protective effects are the
antioxidant nutrients. It is important to note that most of the investigations regarding
inhibitory effect of food component on the oxidative damages of biomolecules have
been devoted to the foods of plant origin.
Oxidation reactions are of major significance in human physiology as well as the
food industry. In the former, oxidative stress has been linked to diseases while food
industry long been concerned with issues such as rancidity and oxidative spoilage of
fruits and vegetables, beverages and other food stuffs (Shahidi, P.; Wanasundara,
P.K. (1992). Grit. Rev. Food Sc. Nutr, 32, 67-103). The widely used food
antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene
(BHT), tertbutylhydroxyquinone (TBHQ) and alkyl gallates are synthetic compounds
of phenolic nature. Synthetic antioxidants, like BHA and BHT, have restricted use in
foods as these synthetic antioxidants are suspected to be carcinogenic (Madhavi, D.
L; Salunkhe, D. K. Toxicological Aspects of Food Antioxidants, In-Foocf
Antioxidants, Eds.; Madhavi, D. L., Deshpande, S. S. Salunkhe, D. K., Marcel
Dekker Inc, New York, 1995, p.267). Consumer concerns and government
regulations are fostering the research on alternative ingredients, mostly natural
polyphenols, as presumably safer food antioxidants with added beneficial functional
properties. Thus the discovery of new natural antioxidants has obvious nutritional
and pharmaceutical relevance.
Garcinia (Family: Guff/ferae) is a large genus of polygamous trees or shrubs,
distributed in the tropical Asia, Africa and Polynesia. It consists of 180 species, out of
which about 30 species are found in India (The Wealth of India -Raw Materials,
Vol.IV, CSIR, New Delhi, India, 1956, pp.99-108). Among the species Garcinia cowa
is found in Northeastern part of India and Andaman Islands. In upper Assam, it is
often cultivated in homesteads for its acid fruit. The fruits are edible, dried slices of
the fruit are used in curry in place of tamarind/lemon. Garcinia is a rich source of
secondary metabolites including xanthones, flavonoids, benzophenones, lactones
and phenolic acids. Since most xanthones have phenolic functional groups on a
linear tricyclic ring they often exhibit a wide range of biological and pharmacological
activities (Bennet, G.J.; Lee, H.H. Xanthones from Guttiferae. Phytochemistry, 28,
967-998, 1989; Minami, H., Kinoshita, M., Fukuyama, Y., Kodama, M., Yoshizawa,
T., Sugiura, M. Nakagawa, K. and Tago, H. 1994, Antioxidant xanthones from
Garcinia subelliptica. Phytochemistry 36, 501-506).
In the background of synthetic antioxidants being used in foods, there is a
need to explore the possibility of isolation of antioxidants from plant sources.
To overcome this drawback, the present invention provides a process for
preparation of antioxidants from Garcinia, wherein the process relates to the
preparation of antioxidants from the rinds of Garcinia cowa.
The main object of the present invention is to provide a process for the
preparation of antioxidants from Garcinia, wherein rinds Garcinia cowa are used in
the preparation of natural antioxidants.
An object of the present invention is to use simple extraction methods and
solvents used for the preparation of antioxidants.
Another object of the present invention is to produce the antioxidants, which is
essentially neutral in flavour and dark brown in colour. The process is efficient and itinvolves the use of simple extraction methods and solvents, which can be regenerated.
Accordingly, the present invention provides a process for the preparation of xanthon antioxidants from Garcinia, which comprises : i) selecting the rinds of Garcinia cowa ii) characterized in that cutting the rinds of Garcinia cowa manually
to a size of 4x6 mm to 8x10 mm, iii) extracting of the above said material (ii) with non polar solvent in
a Soxhlet extractor at temperature in the range of 55-60°C for a
period of 4-8 hr, iv) filtering the above extract to obtain the particle free extract, v) concentrating the above particle free extract at a temperature of
35-40°C under vacuum may be effected to recover the solvent
to an extent of 95%, vi) drying the above concentrated extracted using vacuum oven at
40-45°C under vacuum at 8-27 mm of mercury to obtain
xanthon antioxidant.
In an embodiment of the present invention, the yield of chloroform extract may be in the range of 5.5-8.0%.

In an embodiment of the present invention, the product has antioxidant activity
as potent as or more potent than butylated hydroxyanisole (BHA) is useful in food
and medicine.
The preparation of antioxidants from Garcinia cowa was done according to
following flow diagram:
Rinds of Garcinia cowa
Soxhlet extractor
Non-polar solvents at 55-60 °C
Extract
Concentrated at 35-40°C
and 10-25 mm of mercury
Spent
It can be used for
Hydroxycitric acid extraction
Dried under vacuum at 40-50 °C
and 10-25 mm of mercury
Yield of antioxidant fraction (6.0-7.5%)
Recovery of solvent
The novelty of the process is
1. This is the first report of preparation of antioxidants from G. cowa.
2. The invention is a single step process to obtain the antioxidant fraction
from G. cowa.
The advantages of the process is
1. Preparation of natural antioxidants
2. The process is simple and the solvents used in this process can be
regenerated for further use.
3. The left over spent can be further used for the extraction of (-)-Hydroxycitric
acid.
The following examples are given by way of illustration of the present
invention and therefore should not be constructed to limit the scope of the present
invention.
Example -1
Fifty grams of rinds of Garcinia cowa was manually cut into small pieces and
extracted using 200 ml of cyclohexane at 60 °C for 4 h in a Soxhlet extractor. The
extract was filtered using Whatman filter paper No.1. The extract was concentrated
at a temperature of 40 °C and under redyced pressure at 10 mm of mercury to
recover the solvent and dried in vacuum oven at 40 °C and 25 mm of mercury. The
yield of antioxidant extract was 3.1 g.
Example -2
The dried rinds in 100 g aliquots of Garcinia cowa were extracted with 400 ml
of hexane by using Soxhlet extractor at 55 °C for 8 h. The extract was collected
and filtered through Whatman filter paper no. 1. The filtrate was concentrated at a
temperature of 35 °C and under a reduced pressure at 25 mm of mercury to recover
the solvent and dried in vacuum oven at a temperature of 50 °C and under vacuum
of 10 mm of mercury. The yield of extract was 6.4 g.
The extract obtained from the above method was screened for
antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) (Singh et al., J.
Agricultural and Food Chemistry, 50, 81-86, 2002). At 10 ppm concentration, both
BHA and the extract of G. cowa showed 76 and 75.7% respectively, free radical
scavenging activity.
Example -3
Rinds in quantities of 150 g of Garcinia cowa were extracted with 600 ml of
benzene by using Soxhlet extractor at 60 °C for 8 h. The extract was filtered
through Whatman filter paper no. 1. The filtrate was concentrated at a temperature
of 40 °C and under reduced pressure at 10 mm of mercury to recover the solvent
and dried in a vacuum oven at 40 °C under vacuum at 25 mm of mercury. The yield
of benzene extract was 8.2 g.
The benzene extract obtained from the above method was screened for
antioxidant activity using (3-carotene-linoleate model systems. BHA and benzene
extract showed 98 and 93.7% antioxidant activity at 200 ppm using (3-carotenelinoleate
method (Jayaprakasha G.K. and Jaganmohan Rao L. Phenolic constituents
from lichen Parmotrema stuppeum (Nyl.) Hale and their antioxidant activity,
Zeitschrift fur Naturforschung, 55c, 1018-1022, 2000).

We claim :
1. A process for the preparation of xanthon antioxidants from Garcinia,
which comprises :
i) selecting the rinds of Garcinia cowa
ii) characterized in that cutting the rinds of Garcinia cowa manually
to a size of 4x6 mm to 8x10 mm, iii) extracting of the above said material (ii) with non polar solvent in
a Soxhlet extractor at temperature in the range of 55-60°C for a
period of 4-8 hr, iv) filtering the above extract to obtain the particle free extract, v) concentrating the above particle free extract at a temperature of
35-40°C under vacuum may be effected to recover the solvent
to an extent of 95%, vi) drying the above concentrated extract using vacuum oven at 40-
45°C under vacuum at 8-27 mm of mercury to obtain xanthon
antioxidant.
2. A process as claimed in claim 1, wherein the non polar solvent used is hexane, cyclohexane, pentane, benzene.
3. A process for the preparation of xanthon antioxidants from Garcinia substantially as herein described with reference to the examples.

Documents:

346-DEL-2003-Abstract-(28-11-2008).pdf

346-del-2003-abstract.pdf

346-DEL-2003-Claims-(28-11-2008).pdf

346-del-2003-claims.pdf

346-DEL-2003-Correspondence-Others-(28-11-2008).pdf

346-del-2003-correspondence-others.pdf

346-del-2003-correspondence-po.pdf

346-DEL-2003-Description (Complete)-(28-11-2008).pdf

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

346-del-2003-form-1.pdf

346-del-2003-form-18.pdf

346-DEL-2003-Form-2-(28-11-2008).pdf

346-del-2003-form-2.pdf

346-DEL-2003-Form-3-(28-11-2008).pdf

346-del-2003-form-3.pdf

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Patent Number

236157

Indian Patent Application Number

346/DEL/2003

PG Journal Number

41/2009

Publication Date

09-Oct-2009

Grant Date

05-Oct-2009

Date of Filing

21-Mar-2003

Name of Patentee

COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH

Applicant Address

RAFI MARG, NEW DELHI-110 001, INDIA

Inventors:

#	Inventor's Name	Inventor's Address
1	GUDDADARANGAVVANAHALLY KRISHNAREDDY JAYAPRAKASHA	CENTRAL FOOD TECHNOLOGICAL RESEARCH INSTITUTE, MYSORE, INDIA
2	BHABANI SANKAR JENA	CENTRAL FOOD TECHNOLOGICAL RESEARCH INSTITUTE, MYSORE, INDIA
3	MANDYAM CHAKRAVARATHY VARADARAJ	CENTRAL FOOD TECHNOLOGICAL RESEARCH INSTITUTE, MYSORE, INDIA

PCT International Classification Number

C09K 15/00

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA