Title of Invention

AN IMPROVED PROCESS FOR EXTRACTION OF PIPERINE FROM PIPER SPECIES

Abstract

All improved process for extraction of piperine of formula 1 from piper species (Formula Removed) i. contacting the piper species with an aqueous hydrotrope solution at temperature from 0-100 0C and separating the solution obtained from the solid residue knov/n methods. ii. Recovering the piperine from the solution obtained at the end of step (1) by solvent extraction with organic solvent after dilution with water or without dilution and then desolventising to recover piperine in pure from or so as to bring the concentration of hydrotrope sufficiently low to precipitate piperine from the solution in solid form and separating the precipitated piperine from the solution obtained, followed by washing with water.

Full Text

The present invention relates to an improved process for extraction of piperine from piper species such as Piper nigrum, Piper retrofractum, Piper longum, Piper schmiditii. This method is particularly related to the extraction of piperine using aqueous hydrotrope solutions for the first time. Piperine (C17H1903N) is the major constituent of black pepper (Piper nigrum) and is used extensively as a spice, condiment, insecticide and for medicinal purposes. It has shown potent chemoprotective effect against procarcinogens (Reen and Rashmet, J. Ethnopharmacol., 1997, 58(3), 165-173). The extracts of Piper nigrum are found to have a hypercoagulative effect in vitro; they lessen the clotting time by accelerating the thrombin activation and lowering the heparin level in the clotting systems. (Hasselstorm et.al., Food Res., 1954 19, 373). They are valued for their rubefacient properties and hence used as local application for sore throat, piles and some skin diseases (Wealth of India, Raw Materials, Vol.Ill, pg.110). The concentration of piperine is about 6% to 9% in Piper nigrum, 4.0% in Piper logum fruits and 4.5% in Piper retrofractum. Prior art for separation of Piperine H. Staudinger and H. Schneider [Ber; 56, 699(1923)] described a process for the extraction of piperine from Piper nigrum fruits, wherein the alkaloid was extracted using ethanol and the extract was treated with aqueous sodium hydroxide to remove all the resinous matter. Other solvents used for extraction are petroleum ether and dichloromethane. The extraction of piperine into thee organic solvents is not selective as other compounds like gums, polysaccharides and resins are extracted reducing the purity of piperine. The post extraction processing to purify piperine is cumbersome and uneconomical. A liquid pepper compositin containing pepper in a concentrated form has been prepared by extracting the pepper fruits with acetone and subsequent treatment with anhydrous lactic acid (Wealth of India, Raw Materials, Vol.lll, pg.lll). Vidal, J.P. and Richard, H., (Scientific Aliments 1987, 7(3), 481-98) outlines a procedure for the production of black pepper oleoresins by dense carbon dioxide extraction or carbon dioxide + ethanol extraction. The extract contains 16.7% essential oils and 42.2% piperine which was very close to that obtained by dichloromethane is solvent, Hans j (3,706594 German. Offen. DE 28/02/1987, Bevffet) described a proeas wherein gaseous carbon dioxide under supercritical conditions is passed downward through a packed bed of pulverised pepper and the loaded solvent is then expanded in I downstream separator for release of the extracted material. However, this process using supercritical fluid extraction is not economical and still a further solvent extraction step|is essential for obtaining pure piperine. Most of these prior art processes relate to the extraction of Piper nigrum oleoresin containing piperine and other volatile oils, Piperine containing oleojresin is then solvent extracted and recrystallised to obtain pure piperine. These conventional processes are carried out using a number of steps which often present difficult operating conditions and result in a high cost of production. - The main object of the present invention is to provide a cost effective process for the extraction of piperine from Piper species in a substantially pure form using the phenomena of hydrotropy The approach was to find a suitable hydrotrope for selecttve extraction of piperine from Piper species followed by dilution of the extract phase to precipitate piperine in pure form. In this process of invention the selective extraction of phytochemicals like piperine is conducted in a simple two - step process which successfully exploits the ability of hydrotropes to dissolve the otherwise water insoluble organic compounds in aqueous solution. The first step involves the selective extraction of piperine from Piper species and its soiubilisatioa in the aqueous hydrotrope solution. The second step involves the recovery of piperine by simple dilution with water as most of the piperine precipitates out. In another embodiment of the invention the extracted piperine is rfcovered from aqueous hydrotrope solution by extraction with organic solvents fWlowed by de soi-ntisation. Another object of the present invention is to provide a process for the extraction of piperine from Piper nigrum in the first step without using an organic solvent or excessive temperature to retain the purity of piperine. Accordingly, the present invention provides An improved process for extraction of piperine of formula 1 from piper species Formula 1 (Formula Removed) . contacting the piper species with an aqueous hydrotrope solution at temperature from 0- 100°C and separating the solution obtained from the solid residue known methods, ii. Recovering the piperine from the solution obtained at the end of step (I) by solvent extraction with organic solvent after dilution with water or without dilution and then desolventising to recover piperine in pure form or so as to bring the concentration of hydrotrope sufficiently low to precipitate piperine from the solution in solid form and separating the precipitated piperine from the solution obtained, followed by washing with water. In an embodiment of the invention the piperine is recovered from the aqueous solution of hydrotrope obtained in step (I), after dilution with water so as to bring the concentration of hydrotrope sufficiently low to precipitate piperine from the solution in solid form and separating the precipitated piperine from the solution obtained, followed by washing with water or without dilution, by extraction with organic solvent selected from a group of aromatic hydrocarbons, aliphatic hydrocarbons, ethers, esters, ketones, amides, alcohols or a mixture thereof. In general the process of the invention is described as follows : The dried fruits of piper species such as piper nigrum, Piper longum, Piper retrofractum, Piper schmididtii in pulverized form, preferably in the mesh size of 5 to 300 are brought in intimate contact with an aqueous solution of hydrotrope in the form of a slurry in a stirred vessel or in a column in which the coarse Piper species powder is packed and the solution passes over it. In the process of invention when the contacting is done in the stirred vessed the Piper species powder is added to the aqueous hydrotrope solution. After the aforementioned components are brought together in the mixture usually in the form of slurry the mixture is agitated for a period sufficient for the extraction of pipeline to take place. A typical mixing time is in the range of 15 minutes to 24.00 hours depending upon the concentration of hydrotrope and the speed of agitation. The mixing is conducted at a selected temperature from 0°C -100°C preferably at room tempefature of 30°C. and atmospheric pressure. The Piper species for the extraction of pipeline is selected from the grou$i consisting of Piper nigrum, Piper longiim, Piper retrofractum, Piper schmidtii The term "hydrotrope" indicates short alkyl chain water soluble amphiphillic compounds. The hydrotrope is selected from the group consisting offsodium, potassium calcium, ammonium, magnesium salts of alkyl benzene sulfonates, such as benzane . sulfonate, toluene sulfonate, xylene sulfonate, ethyl benzene sulfonate; styrene sulfonate| pseudocumene sulfonate, mesitylene sulfonate, propyl benzene sulfonate and butyl benzene sulfonate; alkyl polyglycol sulfates and phosphates such as methyl cellosolve sulfate, ethyl cellosolve sulfate, propyl cellosolve sulfate, butyl cellosolve sulfate, pentyt cellosolve sulfate, hexyl cellosolve sulfate and the corresponding phosphates with sodium, potassium, and calcium counterions; methyl diglycol sulfates, ethyl diglycol sulfates, propyl diglycol sulfates, butyl diglycol sulfates, pentyl diglycol sulfates, hexyl diglycol sulfates and phosphates salts with of sodium, potassium, calcium and) ammonium counterions; substituted aromatic carboxylates such as hydroxybenzotaes, toluates, chiorobenzoates, nitrobenzoates, alkyl benzene carboxylates with sodium, potassium, calcium and ammonium counterions; substituted phenates, such as hydroxyphenates, chlorophenates, alkyl phenates, naphthols; naphthalene carboxylates and substituted naphthalene carboxylates such as hydroxy naphthalene carboxylates After the mixing the solid residue is separated from the solution by decantation or filtration or by centrifugation. The separated residue is washed with water and the washing is combined with filtrate or the filtrate is used as such for the recovery of pipeline. The fiitrste is diluted with water to bring the concentration of hydrotrope to low enough to precipitate pipeline from the solution. The dilution is done at a temperature at a range 00-800C preferably in the range 20°-30°C. The precipitated pipeline is separata! from the solution by decantation or filtration or centrifugation. The pipertne is washad with water and dried. The hydrotrope solution recovered during this step is concentrates and recycled. In another embodiment of this invention the filtrate obtained! after the step (1)is subjected to extraction with an organic solvent for recovery of piperite torn the aqueous solution of hydrotrope after dilution of the hydrotrope solution wifi 'water or withaut dilution. The organic solvent selected for the extraction should beimmiscible with water and is selected from a group consisting of aromatic hydrocarbons, aliphatii, hydrocarbons, halogenated hydrocarbons, ethers, esters, alcohols, amides and a mixture thereof. The objects of the invention, its advantages and means of attaining the same are disclosed hereunder in greater detail with reference to the noh - limiting exemplary embodiments of the same. The examples are by way of illustration only and in no way restrict the scoped of the invention. Chemicals used Piper nigrum (Dried fruits of black pepper) Sodium butyl glycol sulphate (50 % aqueous solution, Hulls Germany) Sodium n-butyl benzene sulphonate salt (synthesised in the laboratory) Sodium cumene sulphonate (Navdeep Chemicals PvtXtd.) Sodium xylene-sulphonate (Navdeep Chemicals Pvt.Ltd.) Sodium p-toluene suiphonate(Navdeep Chemicals Pvt.Ltd) Dichloromethane (AR grade) Methanol (HPLC grade) Method of Analysts: High Performance Liquid Chromatography with C18 reverse phase column. Example 1 Whole pepper 20 gms was pulverised to a coarse powder of mesh size# 6 and was added to 100 ml aqueous sodium butyl glycol sulfate of 2.5 gmol/lit concentration and the suspension was stirred vigorously for 2 hours at 30°C. The solution was filtered am then diluted by addition of 310ml waterThe precipitated piperine was dried and analysed for purity. The recovery of piperine was 58.8% (716 mg.) based on the amount if piperinepresent in the dried fruit with a purity of 98%. Example 2 Whole pepper 20 gms was pulverised to a coarse powder of mesh size# 6 and added to 100 ml aqueous sodium butyl glycol sulfate 2.5 of gmol/lit concentration and the suspension was stirred vigorously for 2 hours at 80°C. The solution was filtered and then diluted three times by addition of 310 ml water. The precipitated piperine was driel and then dissolved in dichloromethane and analysed for purity. The recovery of pipermfc was 56.38% (687.mg.) based on the amount of piperine present in the dried fruit with purity of 89%. Example 3 Whole pepper 20 gms was pulverised to a coarse powder oft mesh size# 6 and added to 100 ml aqueous sodium n- butyl benzene sulfonate of 1.0 gmol/lit concentration and the suspension was stirred vigorously for 2 hours at 30°C. The solution was filterec and then diluted by the addition of 2000 ml of water. The precipitated piperine was separated by centrifugation, dried and then dissolved in dichloromethaiie and analysed for purity. The recovery of piperine was 63.92% (0.76g.) based on the apnount of piperine present in the dried fruit with a purity of 97.4%. Example 4 Whole pepper 20 gms was pulverised to a coarse powder of mesh size# 6 and! added to 100 ml aqueous sodium n- butyl benzene sulfonate of 2.0 gmol/Iit concentration and the suspension was stirred vigorously for 2 hours at 30°C. The solution was filtered | and then diluted by addition of 2000 mi. of water. The precipitated piperine was! separated by eentrifugation, dried and then dissolved in dichioromtfhtne and analyst purity. The recovery of piperine was 95.75% (1.14g.) based on the amount of pipenre present in the dried fruit with a purity of 93%. Example 5 Whole pepper 20 gms was pulverised to a coarse powderof mesh size# 72 and added to 100 ml aqueous sodium n- butyl benzene sulfonate of 2.0 gmol/Iit concentration| and the suspension was stirred vigorously for 2 hours at 30°C. The solution was filtered and then diluted by addition of 2000 mi of water. The precipittted piperine was separated by centrifugation, dried and then dissolved in dichlorornethane and analysed for| purity. The recovery, of piperine was 92.14% (1.095g.) based on the amount of piperi|e| present in the dried fruit with a purity of 90.15%. Example 6 Whole pepper 20 gms was pulverised to a coarse powder of inesh size #300 an| added to 100 ml aqueous sodium n- butyl benzene sulfonate 2.0 gmol/Iit concentration and the suspension was stirred vigorously for 2 hours at 30°C. The solution was filtered and then diluted by addition of 2000 ml water. The precipitated piperine was separated! by centrifugation dried and then dissolved in dichloromethane and analysed for purity. The recovery of pijperine was 90.14% (1.07g.) based on the amount of piperine present in the dried fruit with a purity of 89.14%. Example 7 Whole pepper 20 gms was pulverised to a coarse powder of meth size# 6 and was added to 100 ml aqueous sodium n- butyl benzene sulfonate of 2.0 gmol/Iit concentration and was stirred vigorously for 2 hours at 30°C. The solution was filtered and theft extracted with 50 ml. dichloromethane three times. The recovered piperine was analysed for purity. The recovery of piperine was 96.82% (1.15g.) based on the amount of piperine present in the dried fruit with a purity of 95.13%. Example 8 Whole pepper 20 gms was pulverised to a coarse powder of mesh size# 6 and added to 100 ml of aqueous sodium xylene sulfonate 2.0 gmol/lit concentration and was stirred vigorously for 2 hours at 30°C. The solution was filtered and then diluted by addition of 2000 ml water. The precipitated pipeline was separated by centrifugation dried and then dissolved in dichloromethane and analysed for purity. The recovery of piperine was 65.43% (0.77g.) based on the amount of pipeline present in the dried fruet with a purityof 60.41%..... Example 9 Whole pepper 20 gms was pulverised to a coarse powder Of mesh size# 6 and added to 100 ml of aqueous sodium cumene sulfonate 2.0 gmol/lit concentration and-was stirred vigorously for 2 hours at 30°C. The solution was filtered and then diluted by ■--■■. ■ ' ' i addition of 2000 ml water. The precipitated pipeline was separated by centrifugation dried and then dissolved in dichloromethane and analysed for purity. The recovery of pipeline was 72.3% (0.85g.) based on the amount of piperine present in the dried fruet with a purity of 68.00%. i Example 10 Whole pepper 20 gms was pulverised to a coarse powder of mesh size# 6 ant added to 100 ml aqueous sodium p-toluene sulfonate of 2.0 gmol/lit concentration and |h| suspension was stirred vigorously for 2 hours at 30°C. The solution was filtered and the! diluted by addition of 2000 ml water. The precipitated piperine was separated b| centrifugation, dried and then dissolved in dichloromethane and analysed for purity. The recovery of piperine was 49% (0.582g.) based on the amount of pip#riae present in t|MJ dried fruit with a purity of 52.3%. The main advantages of the present invention are :- 1 A simple practically viable method has been provided for the extraction of piperinei 2 The number of steps required for extraction have been reduced apd simple operating conditions are provided which can reduce the cost of production. 3 The hydrotrope solution can be recycled with or without the concentration step f|r further extraction which reduces the cost of chemicais. 4 The process of the present invention is carried out preferentially at tower temperature which is an improvement over the prior art as it-does not degrade pipeline and other chemicals. ._ 5 The first step of the process of the present invention is carried outt without the use on any organic solvent, which retains the purity of piperine.and is an improvement over the prior art. We Claim: 1. An improved process for extraction of piperine of formula 1 from piper species Formula 1 (Formula Removed) Comprises ; 1. contacting the piper species with an aqueous hydrotrope solution at temperature from 0-100°C and separating the solution obtained from the solid residue known methods. II. Recovering the piperine from the solution obtained at the end of step (I) by solvent extraction with organic solvent after dilution with water or without dilution and then desolventising to recover piperine in pure form or so as to bring the concentration of hydrotrope sufficiently low to precipitate piperine from the solution in solid form and separating the precipitated piperine from the solution obtained, followed by washing with water. 2. A process as claimed in claim 1 wherein the piper species used in the step (1) is piper nigrum piper longum, piper retrofactum, piper schmiditii. 3. A process as claimed in claims 1-2 wherein the hydrotrope used in the step (1) is alkyl benzene sulfonates, alkyl poly glycol sulfate or phosphate, substituted benzene carboxylates, substituted phenates, substituted naphthols and substituted naphthalene carboxylates. 4. An improved process as claimed in claims 1-3 wherein the solvent used in step (II) is selected from a group consisting of aliphatic hydrocarbons, aromatic hydrocarbons, ketones, esters, ethers, alcohols, amides and a mixture thereof. 5. An improved process for the extraction of pipehne from piper species such as herein described with reference to the examples accompanying this specification.

Documents:

1238-del-1999-abstract.pdf

1238-del-1999-claims(cancelled).pdf

1238-del-1999-claims.pdf

1238-del-1999-complete specification (granted).pdf

1238-del-1999-correspondence-others.pdf

1238-del-1999-correspondence-po.pdf

1238-del-1999-description (complete).pdf

1238-del-1999-form-1.pdf

1238-del-1999-form-2.pdf

1238-del-1999-form-3.pdf

1238-del-1999-form-4.pdf

1238-del-1999-form-9.pdf

1238-del-1999-petition-138.pdf