Title of Invention

AN IMPROVED PROCESS OF EXTRACTING CHILI (CAPSICUM) OLEORESIN

Abstract

The present invention relates to an improved process of extracting chili {Capsicum) oleoresin. The invention also provides a process of recovering carotenoid pigment with improved recovery. Chili or paprika powder/pellets is treated with an enzyme mixture, which facilitates better release of chili constituents namely pigments and pungency for extraction purposes. A mixture of binary solvents selected for higher efficiency in terms of extractability of chili constituents is employed for extraction in place of conventional single solvents.The solvent mixtures are prepared by mixing by volume 90 to 10 parts of polar solvents like acetone, ethyl acetate or methyl ethyl ketone with 10 to 90 parts of non-polar solvents like hexane or petroleum ether.

Full Text

TITLE: AN IMPROVED PROCESS OF EXTRACTING CHILI (CAPSICUM) OLEORESIN. Filed of the invention ,the present invention relates to a process of extraction of chili (Capsicum annum L) oleoresin of improved yield. The invention also provides a process of recovering carotenoid pigment with improved recovery/ Background and prior art references Chilies are the dried ripe red fruits of the genus capsicum. The two well-known species are Capsicum annum L. and Capsicum frutenscens L. India is the largest producer and exporter of chilies. The estimated export of dry chilies from India was 75,000 MT tonnes valued Rs. 25580 lakhs in 2001-02. Pungency and colour are the two main quality attributes in chilies. The red colour of chilies is due to the presence of carotenoid pigments like capsanthin, (major pigment, 35 %) capsorubin, zeaxanthin, violaxanthin, cryptoxanthin, - carotene etc. These pigments are present in chilies mainly in the esterified form, and to a small extent in non-esterified forms. The oleoresin of chilies is used in many processed foods like sausages, seafood, meat products etc. to impart a reddish taint and pungency. It is also used in chicken feed so that chicken meat and yolk will get an attractive reddish shade. For food colouration, pungency free, high colour content chili oleoresins are desirable. Chili oleoresins as commercially produced vary in their pigment content and the colour value ranges from 30,000 to 1,00,000 units as, measured by the method prescribed by the Essential oil Association of America (EOA). The colour content of the oleoresin is directly proportional to the pigment concentration the raw material used. But preparation of oleoresin with high colour value is desirable. The present invention relates to a process to upgrade the colour value of pungency-free commercial chili oleoresin. There are several patents on chili / paprika colour, relating to the following aspects: i. Methods for alcohol or oil extraction, usually after saponification with alkali (No. 31,522 Israel 1971, No. 76,142,020, Japan 1976, No. 80,118,967, Japan 1980, No. 81,011,960, Japan 1981, No. 82,133,160, Japan 1982, No. 82,180,663, Japan 1982, and 83,173,164, Japan 1984). ii. Pigment production by inter-esterification, No. 79,142,236, Japan 1979). iii. Methods for pigment stabilization or prevention of fading (54,010,568, Japan 1979 No. 55046, 147, Japan 1980, No. 76,112,561, Japan 1976) and iv. Preparation of formulations for food use (81,011,961, Japan 1981). Other patents relating to paprika colour are as follows: i. Method of preventing paprika colour from fading (JP52128924, 1977), ii. Production of paprika colour involving the use of calcium hydroxide and solvent extraction (JP57131259, 1982), iii. Production of paprika dyestuff by treating paprika extract with caustic alkali in presence of acetone (JP61264061,1986), iv. Improvement of paprika colour stability to oxygen and light by blending with carrot extract and browning reaction product of a saccharide and amino acid (JP57031962, 1982), v. Fade preventing agent for paprika pigment by adding a browning reaction product of a saccharide and an amino acid (JP56041259, 1981), vi. Fade inhibition of paprika colour by adding solvent extract of rosemary, sage or mixture thereof (JP57102955, 1982), vii. Manufacture of odourless paprika colouring matter by absorption of alcohol extract of paprika on a resin of porus polymeric structure (JP3045657, 1991), viii. Production of stable deodorise paprika pigment by bringing paprika oleoresin into contact with carbon dioxide in supercritical state in presence a polar solvent (JP2038464, 1990), ix. Production of concentrate paprika pigment by treating paprika with lipase followed by solvent extraction (JP62115067, 1987), x. Method for separating paprika pigment by silica gel chromatography (JP62115068, 1987), xi. Manufacture of paprika oleoresin capsule by treatment with molto dextrin and modified starch (KR9310538, 1993), xii. Method for production of colouring agent and flavouring material from paprika (HU63311.1993), xiii. Process for isolation of flavour and colour from paprika (HU185706, 1985), xiv. Method for obtaining aromatics and dyestuffs from bell peppers involving extraction with a solvent in a supercritical state (HU28336, 1983), xv. Improvement of colour stability by treatment with vegetables and fruits, which contain carotenoids (US5888574, 1999), xvi. Natural food colour composition involving mixture of edible materials including paprika and palm oil, carrot oil or corn gluten oil (US4304792, 1981), xvii. High temperature counter current solvent extraction (US5773095, 1998), xviii. Method of stabilising a spice extractive and a preservative product by using a small amount of metal deactivator such as an edible water soluble salt of ethylene such as diamine tetra acetic acid (GB974322, 1964), xix. Colour stabilised paprika composition using a natural antioxidant / oil soluble ascorbic acid ester (US 19900525340, 19900518, 1991), xx. Preparation of emulsified paprika colour (JP2001252043, 2001), prevention of paprika pigment fading using caffeic acid ferulic acid, chlorogenic acid and some of their esters etc. (JP57117566, 1992) The above patents do not teach or does not provide any clue regarding improved extraction of chili oeloresin and recovery of carotenoid. According to a published paper by Santamaria et al., "Selective enzymemediated extraction of capsaicinoids and carotenoids from chili guajillo puya (Capsicum annum L) using ethanol as solvent" in J. Agric.Food Chem. 2000, 48, 3063- 3067, selective extraction of capsaicinoids and carotinoids from the Mexican chili Guajillo "puya" was studied. Ethanol was used as solvent for the extraction of carotenoids and capsaicin from chili powder treated with commercial enzymes (individually) containing a) pectolytic as well as hemicellulolytic activities, (b) cellulotytic activity, (c) a vide variety of carbohydrase activities d) with pectin esterase and arabanase activities. It was recommended that, pre-treatment of the flour with a commercial enzyme namely Viscozyme L (with a vide variety of carbohydrase activities having 120 units of fungal P- glucanase (FBG) /ml) at 5% level on chili powder helped to get an increased yield of 11% for carotenoid and 7% for capsaicinoids respectively, over control chili powder. In the present patent proposal using two commercial enzymes namely (a) Extrazyme (with declared activity of 7500 pectinase S units and a multi-enzyme complex containing a vide range of carbohydrases, including arabanase, cellulase, p~ glucanase, hemi-cullalase, and xylanase and (b) Energex, in powder or liquid form, with declared activities of 50 - 120 fungal p--glucanase per gram or ml and 5000 to 12000 PSU per gram or ml. The method reported in the publication referred to above, has many disadvantages, which are overcome in the present patent application. 1) As per the paper, the chili powder to water ratio is very high being of the order of 1:50 which needs to be dried before extraction. This step of removal of water in large quantities involves time and energy, which would make the processes uneconomical. In the present patent proposal material to water ratio for enzyme treatment is far less (1:1) which makes the drying step easier and faster 2) Enzyme addition to the chili powder as reported in the paper ranges from 1-5% and actually the authors recommend 5% of the enzyme. In the present patent proposal maximum level required is 1% only. 3) The increase in the yields of carotenoids and capsaicinoids as per the referred paper is 10% for carotenoids and 7% for capsaicinoids over the control batch. In contrast to this in the proposed patent, the increase in yields of carotenoids and capsicinoids is of the order of 24% and 32% respectively, which is very much higher. 4) The solvent used in the publication is aqueous alcohol or pure alcohol. In the proposed patent a mixture of solvents with an advantage of improved extractability for chili constituents namely, carotenoids and capsaicin has been employed 5) Commercial scale adaptation is feasible for the process claimed in the patent application In the spice extraction Industry, chili oleoresin is produced by extraction of chili powder with solvents such as acetone, ethylene dichloride and hexane followed by desolventisation to get a deep red coloured, viscous material called the oleoresin. The oleoresin contains the constituents responsible for colour namely carotenoids, the pungent constituents mainly capsaicin and dihydrocapsaicin, besides other soluble components such as fats and waxes. The present patent aims at enhancing the extractability of chili constituents especially pigments and capsaicinoids by a combination of two techniques namely enzyme treatment and extraction with selected solvent mixtures. Objects of the invention The main object of the present invention is to provide an improved of extracting chili(capsicum) oleoresin. Accordingly, the present invention provides, an improved process of extracting chili(capsicum) oleoresin, which comprises, i) subjecting chili powder/chili flakes to enzyme treatment characterized in that by mixing with an aqueous solution of a commercially available enzyme (trade name Extrazyme) in the range of 0.1-1.0 by weight part for 100 by weight parts of chili powder/flakes by incubating at a pH in the range of 4.0 to 7.0 for a period ranging between 2 and 24 hours at a temperature of 20 to 50°C, ii) drying the enzyme treated chili powder/flakes to mechanical drying to bring down the moisture level of about 5 -12%, iii) powdering the contents of step (ii) into a particle size of about 20-30 mesh, iv) loading the chili powder or its pellets into batch process or a continuous process multiple extraction columns and soaking in a mixture of solvent consisting polar and non-polar solvent as herein described wherein ratio of polar solvent: non-polar solvent used is in the range of 20 to 60: 80 to 40 and the ratio of starting material to the solvent used is in the range of 1:6 to 1:12 for a period ranging between 1 to 3 hours; v) extracting soaked mixture further with solvent mixture as herein described and pooling the extracts; vi)repeating the steps (iv) and (v), pooling the extracts and drying the pooled extracts; and vii)concentrating the pooled extracts under reduced pressure to obtain enriched oleoresin. In an embodiment of the invention, the multienzyme preparation used for treating the chili powder/flakes belongs to the endo-1,3 (4) ß-glucanase class consisting of pectinase, betaglucanase, hemicellulase and pectinase with activity range of 50-120 fungal beta glucanase units per gram and pectanise 5000-12000 psu/g, the enzyme preparation being used at a level of 0.1-100 to 1-100 on the weight of chili powder/flakes, In another embodiment of the invention, paprika powder/flakes is employed as the raw material for oleoresin extraction, In yet another embodiment of the invention, the chili powder/ paprika powder is converted to the form of pellets of size up to 1cm diameter and up to 15 mm length using commercially available pelletizing machine and is loaded into extractor and extracted with solvent mixtures, In yet an another embodiment of the invention, the chili flakes are obtained by drying the commercially available chili, after pre-cleaning, to a moisture level of up to 8 per cent, adopting natural or mechanical drying techniques followed by grinding in a hammer mill to obtain a mixture of pericarp flakes and seeds which is sieved to separate pericarp and seeds from the fine powder of pericarp, followed by subjecting the mixture of pericarp and seeds to air aspiration using a commercially available air aspirator resulting in separation of seeds from the pericarps, the pericarp being the raw material for enzyme treatment, drying, powdering/pellitization and mixed solvent extraction, In yet an another embodiment of the invention, the enzyme treated chili powder/pellet is loaded into an extractor and extracted using solvent mixture where in, the extract drained is recycled through the material in the extractor in order to increase the concentration of the soluble constituents, and the enriched extract is taken for desolventization and getting the oleoresin, In yet another embodiment of the invention, the solvent used for extraction is preferably taken at a level of 20-60 parts from the group consisting of acetone, ethyl acetate, and ethyl methyl ketone, which is mixed with 40-80 parts of hexane/petroleum ether on volume basis, In yet another embodiment of the invention, the enzyme treated chili powder/pellets is loaded into a series of extractors, up to four in number and extracted using a solvent mixture as stated in step d, wherein, the extract drained out from the first extractor is employed as a solvent for the material in second extractor and so on till the last extractor and the enriched extract containing 10-20 % solids is taken for desolventization and getting the oleoresin. The process consists of first subjecting commercially available chili for thorough cleaning to remove any extraneous matter. This is followed by separation of seeds and flakes followed by grinding of the flakes to powder using a suitable grinder. This is followed by treating the pericarp powder with an aqueous solution of an enzyme mixture designed to cause cell wall degradation, and consisting of constituent enzymes namely, cellulose, hemicellulose, amylase, pectinase, arabinase, S-glucanase and xylanase. The enzyme treated chili powder is held at optimum temperature for enzyme action. The chili powder is dried to a moisture level of 15% or 8% depending on whether the powder is subjected to pelletization or extracted as a powder, respectively. The pellets are dried to moisture content of about 8%. The pellets or the powder is loaded in to columns and extracted with binary solvent mixtures drawn from hexane, acetone, ethyl acetate, ethyl methyl ketone and petroleum ether. It has been found that the selected binary solvent mixtures are more efficient for the extraction of chili constituents as compared to the individually pure solvents. This results in reduction of processing time and also lesser use of solvents, besides improved recovery of the principal constituents namely, carotenoid pigments and pungent principles, which are mainly capsaicin and dihydro capsaicin. The extract or miscella is subjected to distillation under controlled conditions to maximaize the recovery of the solvent and minimize the loss of pigments and to produce an oleoresin of improved colour value and capsaicin content. The solvent mixture adsorbed onto the spent chili powder / pellets is recovered by passing steam through the bed of the material and condensing the vapours. In the case of water immiscible solvent mixture, such as ethyl acetate and hexane the aqueous layer can be readily separated by draining out from condensate. In the case of a solvent mixture such as hexane and acetone, hexane is recovered as a separated layer and the acetone in the aqueous layer of the condensate is recovered by fractional distillation. Novelty 1. Chili or paprika powder/pellets is treated with an enzyme mixture, which facilitates better release of chili constituents namely pigments and pungency for extraction purposes. 2. A mixture of binary solvents selected for higher efficiency in terms of extractability of chili constituents is employed for extraction in place of conventional single solvents. 3. The solvent mixtures are prepared by mixing by volume 90 to 10 parts of polar solvents like acetone, ethyl acetate or methyl ethyl ketone with 10 to 90 parts of non-polar solvents like hexane or petroleum ether. The following examples are given by way of illustration of the present invention and therefore should not be construed to limit the scope of the present invention. Example 1 Byadgi variety of dried red chili was procured locally and it was dried at 50°C for two hours in order to bring down the moisture to nearly 5%. The crisp dry chili was passed through a hammer mill with a suitable sieve in order to get a mixture of pericarp flakes and seeds. This mixture was passed through a smaller mesh sieve to get two fractions namely a) bigger flakes of pericarp free of seeds b) a mixture of smaller flakes and seeds. This mixture of smaller flakes and seeds was passed through an air-aspirator, which resulted in two desired fractions namely pure seeds and pericarp. The preicarp contained a small amount of seeds (3-5%). The two pericarp fractions were mixed and was powdered in a hammer mill fitted with 30 mesh sieve to get a powder which was used as the raw material for conversion into pellets and for extraction studies. The powder (30 g) was loaded in a glass column and extracted with a solvent mixture containing 60 parts of Acetone, and 40 parts of Hexane, by volume. After the solvent addition a contact time of 2 hours was given and extract (60 ml) was drained out while simultaneously adding fresh solvent mixture on to the material in the column to keep it soaked. Totally 6 more such extracts were taken after a contact time of 1 hour every time. The extracts were pooled (240 ml) and desolventized in a rota evaporator at atmospheric pressure till nearly 220 ml of the solvent was recovered. Rest of the distillation was carried out under reduced pressure of 20' and a temperature of less than 60°C using a suction pump till solvent was almost completely removed. At the final stages of distillation the unit was connected to a vacuum pump at a vacuum of 26' and a temperature below 80°C for a period of 15 minutes by which time the solvent smell was fully removed from the product (3.8g). The colour value and capsaicin content in this oleoresin 64,100 and 1.54%. A controlled batch of extraction was also carried out under similar conditions as per the solvent mixtures. The oleoresin yield was 3.8 g and its colour value and capsaicin content were 57800 and 1.24% respectively. In another control batch of extraction hexane was used as the solvent under similar conditions. The yield of resin was 3.7g and its colour value and capsaicin content were 59,000 and 0.83% respectively. Example 2 Byadgi variety of dried red chili was procured locally. Reduction of moisture to a level of around 5%, breaking of capsules, sieving and air aspiration were separation of pericarp from seeds was carried out as described in example 1. The flakes were ground using a hammer mill to pass through 30- mesh sieve. A multi enzyme preparation was added as a suspension in water, at the concentration of 0.5% and pH 4.0, to chili pericarp powder (200g batch) and incubated at 37°C for 12 hours. The control sample i.e. without enzyme addition was also maintained under similar conditions of moisture (30%), pH and temperature. After the incubation period both samples were air dried at 50° C using a conventional dryer to bring down the moisture level to about 8%. The treated and control samples, 75gm each, were loaded in to separate glass columns and extracted with selected solvents, at a material to solvent ratio of 1:12. The following solvent mixers namely, Ethyl acetate plus Hexane (60+40) and Acetone plus Hexane mixture (60+40) were used for extraction Recoveries of chili constituents with reference to resin, carotenoid pigments and capsaicin were computed and comparative profile is presented in table. 1. The colour value in the resin and capsaicin in the resin were higher in enzyme treated chili powder in comparison to control. Table-1. Effect of enzyme treatment and extraction with solvent mixtures on the recovery of chili constituents (Table Removed) A: Acetone, H: Hexane, EA: Ethyl acetate Example 3 Byadgi variety of dried red chili was procured locally. The operations of reduction of moisture to a level of around 5%, breaking of capsules, sieving and air aspiration for separation of pericarp from seeds were carried out as described in example 1. The flakes were ground using a hammer mill to pass through 30-mesh sieve. Chili pericarp powder was conditioned for pelletization by uniformly mixing with water added at 8% by weight. The conditioned powder was charged to the pelletizing machine fitted with a die of 6 mm diameter. The pellets obtained were dried to a moisture level of 8% using a cross-flow drier. A twenty grams of the pellets were loaded into glass columns and extracted separately with the following solvents and solvent mixtures namely, Hexane, Acetone, Ethyl acetate, Acetone + Hexane (60+40) and Ethyl acetate + Hexane (60+40) at a material to solvent ratio of 1:12. The relative extractability with reference to resin and carotenoid pigments is presented in Table 2. Table 2. Effect of Solvent Composition on The Recovery of Resin And Pigments from Chili Pericarp (6 Mm Pellets) (Table Removed) The recoveries of resin and pigment pure higher for the stated solvent mixtures than for pure solvents the hexane, acetone or ethyl acetate. Advantages of this process are 1. There is increased recovery of color and pungency from chili powder and pellets. 2. The prepared oleoresin is of higher colour value and capsaicin content. 3. There is reduction in solvent requirement and also in processing time for making the We claim: 1. An improved process of extracting chili(capsicum) oleoresin, said process comprising the steps of: i) subjecting chili powder/chili flakes to enzyme treatment characterized in that by mixing with an aqueous solution of a commercially available enzyme (trade name Extrazyme) in the range of 0.1-1.0 by weight part for 100 by weight parts of chili powder/flakes by incubating at a pH in the range of 4.0 to 7.0 for a period ranging between 2 and 24 hours at a temperature of 20 to 50°C, ii) drying the enzyme treated chili powder/flakes to mechanical drying to bring down the moisture level of about 5 -12%, iii) powdering the contents of step (ii) into a particle size of about 20-30 mesh, iv) loading the chili powder or its pellets into batch process or a continuous process multiple extraction columns and soaking in a mixture of solvent consisting polar and non-polar solvent as herein described wherein ratio of polar solvent: non-polar solvent used is in the range of 20 to 60: 80 to 40 and the ratio of starting material to the solvent used is in the range of 1:6 to 1:12 for a period ranging between 1 to 3 hours; v) extracting soaked mixture further with solvent mixture as herein described and pooling the extracts; vi) repeating the steps (iv) and (v), pooling the extracts and drying the pooled extracts; and vii) concentrating the pooled extracts under reduced pressure to obtain enriched oleoresin. 2. A process as claimed in claim 1, wherein, the enzyme used is the commercially available endo-1, 3 (4) P-glucanase class enzymes selected from a group consisting of pectinase, betaglucanase, hemicellulase and pectinase with activity range of 50-120 fungal betaglucanase units per gram and pectanise 5000-12000 psu/g. 3. A process as claimed in claim 1, wherein the polar solvent used is selected from a group consisting of ketonic solvents selected from acetone, isobutyl ketone, methyl-ethyl ketone, and ethyl acetate. 4. A process as claimed in claim 1, wherein the non-polar solvent used is selected from a group consisting of hexane, petroleum ether. 5. A process as claimed in step (v) of claim 1, wherein the solvent mixture used is ethyl acetate: hexane or acetone: hexane. 6. An improved process of extracting chili(capsicum) oleoresin substantially as herein described.

Documents:

429-DEL-2003-Abstract-(19-02-2009).pdf

429-del-2003-abstract.pdf

429-DEL-2003-Claims-(08-04-2009).pdf

429-DEL-2003-Claims-(19-02-2009).pdf

429-del-2003-claims.pdf

429-DEL-2003-Correspondence-Others-(05-03-2009).pdf

429-DEL-2003-Correspondence-Others-(08-04-2009).pdf

429-DEL-2003-Correspondence-Others-(19-02-2009).pdf

429-del-2003-correspondence-others.pdf

429-del-2003-correspondence-po.pdf

429-DEL-2003-Description (Complete)-(08-04-2009).pdf

429-DEL-2003-Description (Complete)-(19-02-2009).pdf

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

429-del-2003-form-1.pdf

429-del-2003-form-18.pdf

429-del-2003-form-2.pdf

429-DEL-2003-Form-3-(19-02-2009).pdf

429-del-2003-form-3.pdf

429-DEL-2003-Petition-137-(05-03-2009).pdf