Title of Invention	AN IMPROVED PROCESS FOR THE EXTRACTION OF TOTAL SANTALINS FROM THE HEARTWOOD OF PLANTS PTEROCARPUS SANTALINUS L.
Abstract	The present invention provides a simple economical process for the isolation of total santalins from the red sanders wood (Pterocarpus santalinus L). By the process of the present invention total santalins can be separated in powder form wherein the powder contains 61.05 to 79.91% santalins. In the present process no organic solvents are employed to separate and purify the total santalins from red sanders and the yield of santalin from the heartwood of said red sanders ranges from 8-14%.

Title of Invention

AN IMPROVED PROCESS FOR THE EXTRACTION OF TOTAL SANTALINS FROM THE HEARTWOOD OF PLANTS PTEROCARPUS SANTALINUS L.

Abstract

The present invention provides a simple economical process for the isolation of total santalins from the red sanders wood (Pterocarpus santalinus L). By the process of the present invention total santalins can be separated in powder form wherein the powder contains 61.05 to 79.91% santalins. In the present process no organic solvents are employed to separate and purify the total santalins from red sanders and the yield of santalin from the heartwood of said red sanders ranges from 8-14%.

Full Text	This invention relates to an improved process for the isolation of total santalins from the heartwood of Pterocarpus santalinus L. (Red sandes) Red sanders, Pterocarpus santalinus (Fabaceae) is found in dry deciduous forests of Andhra Pradesh mostly in Cuddapah district. This species is also found in small pockets in adjoining Tamil Nadu State. The timber is hard and durable. It is used for agriculture implements for poles, shifts and bent rims of carts and other joinery purposes. It is exported to Japan where it is used in the manufacture of musical instrument called Shamisen. Red sanders wood mainly contains a red colouring matter , santalin (Santalic acid) possessing a quinonoid structure. Santalin yields a blood red solution with alcohol, yellow with ether, and violet with ammonia and caustic alkalies, but is insoluble in water. A variety of compounds including isoflavones stilbenes, pterocarpans and terpenes have been reported from the wood { Seshadri, T.R. (1972) Phytochem. 3,881}. The light petroleum ether extract contains (3-eudesmol (Parthasarthy, M.R. and Seshadri T.R. (1965) Curr. Sci. 34, 115}. The benzene extract is reported to contain pterostilbene, acetyl oleanolic aldehyde, cryptomeridiol, acetyl oleanolic acid, pterocarpol and isopterocarpolene {N. kumaretal (1974) Phytochem. 13,633-636}. The ground wood is chiefly used for dyeing wool, cotton, leather and for staining other woods. The dye is also used for colouring pharmaceutical preparations, foods stuffs and is suitable for colouring pulp. A histological stain has been prepared from the alcohol soluble fraction of the heartwood. The bark is sometimes used in the curing of arecanuts { Ann. (1969) wealth of India, Vol. 8, New Delhi, India , Publications and information Directorate, CSIR 305 -307}. The wood has been used for curing inflammations and head ache, as antipyretic, anthelmintic, aphrodisiac, alexeteric and in billiousness, mental aberrations and ulcers. An infusion of the wood has been used in the control of diabetes. (Kirtikar K.R.., Basu B.D. Indian medicinal plants vol. 1 Indian, Periodical Experts 1981 : 826 ). Large quantities of Red sanders wood are said to have been exported regularly from this country to Europe, where it is employed for extraction of dye. Red sanders is exported to Japan, Hongkong, Germany and Ceylon. Exports of red sandal powder from India averaged 50 tonnes annually between 1988-1993 . The major importers have been Japan, China (Taiwan province) and western Europe . Only the heartwood is employed as a source of the colourant. The process involves reduction of the wood to chips or powdering and using as such. No reliable published information is available on commercial extraction audits yield. For sale to end users, specific formulations (as liquid, dispersed solids or water soluble forms) are prepared at strengths appropriate for the food product. The pigment of red sandal wood and of related red woods have attracted the interest of natural product chemists for more than a century since the first report by Pelletier ( Pelletier., J., Annalen, 1833, 6 : 28 ) Despite much effort, it was not until 1954 that two pure compounds santalin and santarubin were isolated by Roberton and Whalley ( Robertsen, A., and Whalley, W.B., ( 1954 ) J. Chem. Soc. 2794 ) . The presence of two pigments , santalin A and B were reported by Ravindranath and Seshadri ( Ravindranath, B and Seshadri, T.R., ( 1972 ) Tet. Letts 1201 ). Santalin A, B and C were reported by Arnone, et al ( Arnone, A. et al ( 1975 ) J.C.S Perkin I, 186 ). Arnone isolated the santalin by extracting the heartwood shavings with hexane, then with chloroform and ethyl acetate. The ethyl acetate extract was concentrated and the residue chromatographed through a woolen polyamide column (CHCl3-MeOH) to give red products of which santalins A and B were the major constituents. Kumar, N. et al., [Kumar , N et al (1975) J.C.S. Perkin I, 186] separated mixture of santalin A and B alongwith one major and minor constituent by extracting the sapwood shavings with light petroleum, chloroform and ethanol in succession. Chloroform and ethanol extracts were combined and solvent removed. The residue dissolved in hot benzene and left overnight in the refrigerator. The mother liquor after filtration of the content was enriched with santalin. This was chromatographed over polyamide column to obtain santalins. [Kumar, N., Seshadri, T.R., (1974) Curr. Sci., 43, 611]. Robertson & Whalley isolated santalins by extracting the powdered sandalwood with carbon tetrachloride followed by diethyl ether. Ether extract after evaporating the solvent, dissolved in ethyl acetate which on keeping the content as such gave a solid. This solid on crystallization from alcohol yielded santalins. (Robertson, A., and Whalley, W.B. (1954) /. Chem. Soc. 2794]. The methods described in the literature has following disadvantages: 1. No published information is available on commercial extraction and its yields. 2. Ternary and quaternary mixtures of solvents have been employed for extraction of santalins. 3. Estimation and quantitation of total santalin has not been determined in the product. Santalin has been separated by using polyamide column. 4. Summary of basic information on red sandal wood, rated the availability of reliable published information as poor. 5. For the food industries, the extract is normally sold in the alcohol soluble form either as liquids or powders. The purification of the extract has not been reported in the literature. Present status initiated us to develop a simple and economical process for the isolation of total santalins without resorting to chromatography procedures. However, applications of this concept was not obvious, without carrying out the experiments. The main object of the present invention is to provide a simple economical process for the isolation of total santalins from the red sanders wood. Another object is to provid a process by which total santalins can be separated in powder form. Yet another object is to provide a process in which no organic solvents are employed to separate and purify the total santalins from red sanders. Accordingly, the present invention provides an improved process for the preparation of total santalins from the heartwood of the plant Pterocarpus satalinus ( Red sanders wood) which comprises: a) pulverizing the heart wood of red sanders, b) extracting the powdered red sanders in an alkaline medium at a temperature in the range of 10 to 100°C c) filtrating the content obtained in step (b) to get extract and washing the mare with water, d) acidifying the said extract with acidifying agent selected from , organic acids, mineral acids or any weak acids selected from CO2 , CH3COOH,oxalic acid , tartaric acid. e) keeping the acidified mixture below room temperature in a refrigerator for precipitation, f) filtrating and washing of the precipitate with water, g) drying of the precipitate under reduced pressure to get santalins in the form of powder having yield as herein described . hi an embodiment of the process the extraction of the powdered red sanders as described in step (b) may be effected with sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, liq. ammonia or in any alkaline medium and their mixtures. In another embodiment of the process the extraction of the red sanders as described in step (b) can be effected at temperature. hi yet another embodiment of the process the extraction time as described in step (b) can be varied from 2 min to 24 hours. In yet another embodiment of the process the pH of the extract may be changed in the range of 4.0-6.9 with any organic acid, mineral acid or any weak acids selected from CO2.CH3COOH, oxalic acid, tartaric acid . hi yet another embodiment of the process the precipitate may be dried using any drying condition normally applied. Total santalins prepared by the present invention has the following characteristics: Physical appearance : Dark reddish powder Solubility : Soluble in Ethyl alcohol, DMSO, Butanol etc. Solubility in water : NIL Total santalins as assayed spectrophotometricaly 61.05 to 79.91% Yield : 8-14% The details of the process is described herein with the help of examples are by the way of illustration only and therefore should not be construed to limit the scope of invention Example 1 : 50 gm P. santalinus powdered heartwood taken in a conical flask and 1% sodium hydroxide (400 ml) solution added to it . The contents stirred frequently and filtered after 18 hours. The plant marc washed with distilled water (400 ml). The pH of the combined filtrate brought to 6.5 by adding 2% aq. tartaric acid. The contents kept in the refrigerator for 1 hour and filtered. The precipitate washed with water and dried to afford 6.0 gm powder (12% yield) having total santalins 76.08%. Example 2 : 10 gm of P. santalinus powdered heartwood stirred with 14% aq. ammonia (80 ml) for 1 hour, at room temperature and filtered. The marc stirred again with water (50 ml.) and filtered. The combined filtrate cooled and acidified by bubbling carbon-dioxide gas. The acidified contents kept in a refrigerator for 1 hour and filtered. The precipitate washed with water, dried in vacuum to obtain 0.92 gm, powder (yield 9.2%), total santalins in the powder= 79.91%. Example 3 : 100 gm of P. santalinus powder extracted with 2% aq. pot. hydroxide (500 ml) by shaking at room temperature for 2 hours. The contents filtered and marc washed with water (200 ml) thrice. The pooled extract kept in the refrigerator overnight and was acidified with 30% CH3COOH to pH 6.5. The precipitate filtered, washed thoroughly with water and dried to afford 13.90 gm (13.90% yield) solid which was assayed for total santalins as 62.5%. Example 4 : 50 gm P. santalinus powder extracted at room temp, by stirring with 300 ml. of sodium carbonate solution (10%) for 15 minutes. The contents filtered and the plant marc washed with 300 ml. water and washings pooled into the filtrate. Through the combined filtrate carbon dioxide gas was passed for 20 minutes. The separated ppt. was filtered, washed with water, dried under reduced pressure. Weight of the powder = 4.00 gm. (8% yield). Total santalins assayed = 62.98%. The main advantages of the present process are : 1. A process for the extraction of total santalins from the heart wood of P. santalinus by extraction in alkaline medium. 2. A process for the isolation of a powdered material with santalin content varying from 61.05 to 79.91%. 3. Chromatographic techniques are not involved. 4. No use of organic solvents. We Claim: 1. An improved process for the preparation of total santalins from the heartwood of plant Pterocarpus santalinus L ( Red sanders wood) which comprises: a) pulverizing the heart wood of red sanders, b) extracting the powdered red sanders in an alkaline medium at a temperature in the range of 10 to 100°C c) filtrating the content obtained in step (b) to get extract and washing the mare with water, d) acidifying the said extract with acidifying agent selected from , organic acids, mineral acids or any weak acids selected from CO2 , CH3COOH,oxalic acid , tartaric acid , e) keeping the acidified mixture below room temperature in as refrigerator for precipitation, f) filtrating and washing of the precipitate with water, g) drying of the precipitate under reduced pressure to get santalins in the form of powder having yield as herein described, 2. An improved process as claimed in claim 1 wherein the alkaline medium used for extraction is selected from weak alkaline medium to strong alkaline medium of the pH ranging from 8-11. 3. An improved process as claimed in claims 1-2 wherein the stirring of mixture is effected at room temperature. 4. An improved process as claimed in claims 1 - 3 wherein the drying of precipitate of step h is effected under vaccum and kept away from light. 5. An improved process as claimed in claims 1-4 wherein the powder obtained at step (h) has 61.05 to 79.91% total santalins. 6. An improved process as claimed in claims 1 - 5 wherein the yield of santalins from heartwood of Pterocarpus species is in the range of 8 - 14% . 7. An improved process as claimed in claims 1 - 6 by wherein the powder obtained can be used as a dye for pharmaceutical preparation, food stuffs, cosmetic, textile or as solar screen. 8. An improved process for the extraction of total santalins from the heartwood of plants Pterocarpus santalinus L. substantially as herein described with reference to examples.

Full Text

This invention relates to an improved process for the isolation of total santalins from the heartwood of Pterocarpus santalinus L. (Red sandes) Red sanders, Pterocarpus santalinus (Fabaceae) is found in dry deciduous forests of Andhra Pradesh mostly in Cuddapah district. This species is also found in small pockets in adjoining Tamil Nadu State. The timber is hard and durable. It is used for agriculture implements for poles, shifts and bent rims of carts and other joinery purposes. It is exported to Japan where it is used in the manufacture of musical instrument called Shamisen.
Red sanders wood mainly contains a red colouring matter , santalin (Santalic acid) possessing a quinonoid structure. Santalin yields a blood red solution with alcohol, yellow with ether, and violet with ammonia and caustic alkalies, but is insoluble in water. A variety of compounds including isoflavones stilbenes, pterocarpans and terpenes have been reported from the wood { Seshadri, T.R. (1972) Phytochem. 3,881}. The light petroleum ether extract contains (3-eudesmol (Parthasarthy, M.R. and Seshadri T.R. (1965) Curr. Sci. 34, 115}. The benzene extract is reported to contain pterostilbene, acetyl oleanolic aldehyde, cryptomeridiol, acetyl oleanolic acid, pterocarpol and isopterocarpolene {N. kumaretal (1974) Phytochem. 13,633-636}.
The ground wood is chiefly used for dyeing wool, cotton, leather and for staining other woods. The dye is also used for colouring pharmaceutical preparations, foods stuffs and is suitable for colouring pulp. A histological stain has been prepared from the alcohol soluble fraction of the heartwood. The bark is sometimes used in the curing of arecanuts { Ann. (1969) wealth of India, Vol. 8, New Delhi, India , Publications and information Directorate, CSIR 305 -307}. The wood has been used for curing inflammations and

head ache, as antipyretic, anthelmintic, aphrodisiac, alexeteric and in billiousness, mental aberrations and ulcers. An infusion of the wood has been used in the control of diabetes. (Kirtikar K.R.., Basu B.D. Indian medicinal plants vol. 1 Indian, Periodical Experts 1981 : 826 ). Large quantities of Red sanders wood are said to have been exported regularly from this country to Europe, where it is employed for extraction of dye. Red sanders is exported to Japan, Hongkong, Germany and Ceylon. Exports of red sandal powder from India averaged 50 tonnes annually between 1988-1993 . The major importers have been Japan, China (Taiwan province) and western Europe .
Only the heartwood is employed as a source of the colourant. The process involves reduction of the wood to chips or powdering and using as such. No reliable published information is available on commercial extraction audits yield.
For sale to end users, specific formulations (as liquid, dispersed solids or water soluble forms) are prepared at strengths appropriate for the food product. The pigment of red sandal wood and of related red woods have attracted the interest of natural product chemists for more than a century since the first report by Pelletier ( Pelletier., J., Annalen, 1833, 6 : 28 ) Despite much effort, it was not until 1954 that two pure compounds santalin and santarubin were isolated by Roberton and Whalley ( Robertsen, A., and Whalley, W.B., ( 1954 ) J. Chem. Soc. 2794 ) . The presence of two pigments , santalin A and B were reported by Ravindranath and Seshadri ( Ravindranath, B and Seshadri, T.R., ( 1972 ) Tet. Letts 1201 ). Santalin A, B and C were reported by Arnone, et al ( Arnone, A. et al ( 1975 ) J.C.S Perkin I, 186 ). Arnone isolated the santalin by extracting the heartwood shavings with hexane, then with chloroform and ethyl acetate. The ethyl acetate extract was concentrated and the residue

chromatographed through a woolen polyamide column (CHCl3-MeOH) to give red
products of which santalins A and B were the major constituents.
Kumar, N. et al., [Kumar , N et al (1975) J.C.S. Perkin I, 186] separated mixture of
santalin A and B alongwith one major and minor constituent by extracting the sapwood
shavings with light petroleum, chloroform and ethanol in succession. Chloroform and
ethanol extracts were combined and solvent removed. The residue dissolved in hot
benzene and left overnight in the refrigerator. The mother liquor after filtration of the
content was enriched with santalin. This was chromatographed over polyamide column
to obtain santalins.
[Kumar, N., Seshadri, T.R., (1974) Curr. Sci., 43, 611].
Robertson & Whalley isolated santalins by extracting the powdered sandalwood with
carbon tetrachloride followed by diethyl ether. Ether extract after evaporating the solvent,
dissolved in ethyl acetate which on keeping the content as such gave a solid. This solid
on crystallization from alcohol yielded santalins. (Robertson, A., and Whalley, W.B.
(1954) /. Chem. Soc. 2794].
The methods described in the literature has following disadvantages:
1. No published information is available on commercial extraction and its yields.
2. Ternary and quaternary mixtures of solvents have been employed for extraction of
santalins.
3. Estimation and quantitation of total santalin has not been determined in the product.
Santalin has been separated by using polyamide column.
4. Summary of basic information on red sandal wood, rated the availability of reliable
published information as poor.

5. For the food industries, the extract is normally sold in the alcohol soluble form either as liquids or powders. The purification of the extract has not been reported in the literature.
Present status initiated us to develop a simple and economical process for the isolation of total santalins without resorting to chromatography procedures. However, applications of this concept was not obvious, without carrying out the experiments.
The main object of the present invention is to provide a simple economical process for the isolation of total santalins from the red sanders wood. Another object is to provid a process by which total santalins can be separated in powder form. Yet another object is to provide a process in which no organic solvents are employed to separate and purify the total santalins from red sanders.
Accordingly, the present invention provides an improved process for the preparation of total santalins from the heartwood of the plant Pterocarpus satalinus ( Red sanders wood) which comprises:
a) pulverizing the heart wood of red sanders,
b) extracting the powdered red sanders in an alkaline medium at a temperature in
the range of 10 to 100°C
c) filtrating the content obtained in step (b) to get extract and washing the mare with
water,
d) acidifying the said extract with acidifying agent selected from , organic acids,
mineral acids or any weak acids selected from CO2 , CH3COOH,oxalic acid ,
tartaric acid.
e) keeping the acidified mixture below room temperature in a refrigerator for
precipitation,
f) filtrating and washing of the precipitate with water,
g) drying of the precipitate under reduced pressure to get santalins in the form of
powder having yield as herein described .

hi an embodiment of the process the extraction of the powdered red sanders as
described in step (b) may be effected with sodium hydroxide, potassium
hydroxide, sodium carbonate, sodium bicarbonate, liq. ammonia or in any
alkaline medium and their mixtures.
In another embodiment of the process the extraction of the red sanders as
described in step (b) can be effected at temperature.
hi yet another embodiment of the process the extraction time as described in step
(b) can be varied from 2 min to 24 hours.
In yet another embodiment of the process the pH of the extract may be changed
in the range of 4.0-6.9 with any organic acid, mineral acid or any weak acids
selected from CO2.CH3COOH, oxalic acid, tartaric acid .
hi yet another embodiment of the process the precipitate may be dried using any
drying condition normally applied.
Total santalins prepared by the present invention has the following characteristics:
Physical appearance : Dark reddish powder
Solubility : Soluble in Ethyl alcohol, DMSO, Butanol etc.
Solubility in water : NIL
Total santalins as assayed spectrophotometricaly 61.05 to 79.91% Yield : 8-14%
The details of the process is described herein with the help of examples are by the way of illustration only and therefore should not be construed to limit the scope of invention

Example 1 : 50 gm P. santalinus powdered heartwood taken in a conical flask and 1% sodium hydroxide (400 ml) solution added to it . The contents stirred frequently and filtered after 18 hours. The plant marc washed with distilled water (400 ml). The pH of the combined filtrate brought to 6.5 by adding 2% aq. tartaric acid. The contents kept in the refrigerator for 1 hour and filtered. The precipitate washed with water and dried to afford 6.0 gm powder (12% yield) having total santalins 76.08%.
Example 2 : 10 gm of P. santalinus powdered heartwood stirred with 14% aq. ammonia (80 ml) for 1 hour, at room temperature and filtered. The marc stirred again with water (50 ml.) and filtered. The combined filtrate cooled and acidified by bubbling carbon-dioxide gas. The acidified contents kept in a refrigerator for 1 hour and filtered. The precipitate washed with water, dried in vacuum to obtain 0.92 gm, powder (yield 9.2%), total santalins in the powder= 79.91%.
Example 3 : 100 gm of P. santalinus powder extracted with 2% aq. pot. hydroxide (500 ml) by shaking at room temperature for 2 hours. The contents filtered and marc washed with water (200 ml) thrice. The pooled extract kept in the refrigerator overnight and was acidified with 30% CH3COOH to pH 6.5. The precipitate filtered, washed thoroughly with water and dried to afford 13.90 gm (13.90% yield) solid which was assayed for total santalins as 62.5%.
Example 4 : 50 gm P. santalinus powder extracted at room temp, by stirring with 300 ml. of sodium carbonate solution (10%) for 15 minutes. The contents filtered and the plant marc washed with 300 ml. water and washings pooled into the filtrate. Through the

combined filtrate carbon dioxide gas was passed for 20 minutes. The separated ppt. was
filtered, washed with water, dried under reduced pressure.
Weight of the powder = 4.00 gm. (8% yield). Total santalins assayed = 62.98%.
The main advantages of the present process are :
1. A process for the extraction of total santalins from the heart wood of P. santalinus by
extraction in alkaline medium.
2. A process for the isolation of a powdered material with santalin content varying from
61.05 to 79.91%.
3. Chromatographic techniques are not involved.
4. No use of organic solvents.

We Claim:
1. An improved process for the preparation of total santalins from the heartwood of plant Pterocarpus santalinus L ( Red sanders wood) which comprises:
a) pulverizing the heart wood of red sanders,
b) extracting the powdered red sanders in an alkaline medium at a
temperature in the range of 10 to 100°C
c) filtrating the content obtained in step (b) to get extract and washing the
mare with water,
d) acidifying the said extract with acidifying agent selected from , organic
acids, mineral acids or any weak acids selected from CO2 ,
CH3COOH,oxalic acid , tartaric acid ,
e) keeping the acidified mixture below room temperature in as refrigerator for
precipitation,
f) filtrating and washing of the precipitate with water,
g) drying of the precipitate under reduced pressure to get santalins in the
form of powder having yield as herein described,

2. An improved process as claimed in claim 1 wherein the alkaline medium
used for extraction is selected from weak alkaline medium to strong alkaline
medium of the pH ranging from 8-11.
3. An improved process as claimed in claims 1-2 wherein the stirring of
mixture is effected at room temperature.
4. An improved process as claimed in claims 1 - 3 wherein the drying of
precipitate of step h is effected under vaccum and kept away from light.

5. An improved process as claimed in claims 1-4 wherein the powder obtained
at step (h) has 61.05 to 79.91% total santalins.
6. An improved process as claimed in claims 1 - 5 wherein the yield of
santalins from heartwood of Pterocarpus species is in the range of 8 - 14% .
7. An improved process as claimed in claims 1 - 6 by wherein the powder
obtained can be used as a dye for pharmaceutical preparation, food stuffs,
cosmetic, textile or as solar screen.
8. An improved process for the extraction of total santalins from the heartwood
of plants Pterocarpus santalinus L. substantially as herein described with
reference to examples.

Documents:

1101-del-2001-abstract.pdf

1101-del-2001-claims.pdf

1101-del-2001-correspondence-others.pdf

1101-del-2001-correspondence-po.pdf

1101-del-2001-description (complete).pdf

1101-del-2001-form-1.pdf

1101-del-2001-form-18.pdf

1101-del-2001-form-2.pdf

1101-del-2001-form-3.pdf

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Patent Number

227103

Indian Patent Application Number

1101/DEL/2001

PG Journal Number

04/2009

Publication Date

23-Jan-2009

Grant Date

01-Jan-2009

Date of Filing

31-Oct-2001

Name of Patentee

COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH,

Applicant Address

RAFI MARG, NEW DLHI-110001,

Inventors:

#	Inventor's Name	Inventor's Address
1	SATINDER MOHAN JAIN,	India Jammu & Kashmir India
2	GHULAM NABI QAZI,	India Jammu & Kashmir India
3	OM PARKASH SURI	India Jammu & Kashmir India
4	DEVINDER KUMAR GUPTA,	REGIONAL RESEARCH LABORATORY (CSIR), JAMMU

PCT International Classification Number

A61Q19/04; A61K8/49H2

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA