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WO2006103515A1 - Antioxidant conserve from green coffee and process for its preparation - Google Patents

Antioxidant conserve from green coffee and process for its preparation Download PDF

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Publication number
WO2006103515A1
WO2006103515A1 PCT/IB2006/000681 IB2006000681W WO2006103515A1 WO 2006103515 A1 WO2006103515 A1 WO 2006103515A1 IB 2006000681 W IB2006000681 W IB 2006000681W WO 2006103515 A1 WO2006103515 A1 WO 2006103515A1
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WO
WIPO (PCT)
Prior art keywords
flakes
coffee
conserve
solvent
enzyme
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PCT/IB2006/000681
Other languages
French (fr)
Inventor
Madeneni Madhava Naidu
Sathyagalam Ranganatha Desikacharya Sampathu
Bashyam Raghavan
Kesthur Venkatesh Murthy
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Council Of Scientific And Industrial Research
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Publication of WO2006103515A1 publication Critical patent/WO2006103515A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/74—Rubiaceae (Madder family)
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
    • A23F5/00—Coffee; Coffee substitutes; Preparations thereof
    • A23F5/02—Treating green coffee; Preparations produced thereby
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • the present invention relates to an improved process for the preparation of antioxidant conserve from green coffee.
  • Coffee is one of the important agricultural commodities and which is cultivated globally on 11.6 million hectare.
  • the productions of the agricultural commodities have been reached approximately up to 63,000,000s tons. Brazil and Colombia are accounted for nearly 44% of the world production for the same.
  • the coffee is grown in about 80 countries across the globe and out of them near about 51 countries is considered to be the major producers for the coffee.
  • This industry is a major source of direct employment for about 4, 00,000 people in the area of cultivation of agro based products. Apart from this, the industry is also providing indirect employment to many people in processing and trade industry or auxiliaries. It is a highly labor-intensive industry, employing an estimated of about 100 million people are working all over 60 developing countries.
  • the green coffee beans yield caffeine as a by product during the coffee decaffeination process.
  • the other reported prior art are related to use of green coffee powder for analytical purposes i.e. the analysis of green coffee constituents such as caffeine, chlorogenic acids etc.
  • coffee is a good source of phenolic compounds, which are known for use as antioxidants, anti-tumor agents and also posses a host of other therapeutic properties.
  • the phenolic acids in coffee are mainly quinic acids with different amount of caffeoyl groups attached to its different positions.
  • the phenolic acids present in coffee are such as chlorogenic acid, caffeic acid, para- coumaric acid and eugenol which have been shown preventive activities to exert cancer in animal models.
  • the chlorgenic acid which is the main phenolic acid in coffee, is able to protect the gastric mucosa against irritation, and therefore, improves the digestibility of foods, beverages and medicaments.
  • the improved digestibility is expressed through a much reduced systemic acid secretion, which has been found to be directly dependent on an increased level of chlorogenic acid content.
  • chlorogenic acid has a chemopreventive effect on rat stomach cancer (Shimizu et ah, 1999).
  • the natural chlorogenic acid content of coffee is reduced by approximately 40 to 80% during roasting process (Cliford et al., 1998).
  • the use of green coffee bean will result in a product, which preserves the phenolic compounds.
  • Green coffee contains a large amount of polyphenols exemplified by chlorogenic acid, caffeic acid, ferulic acid and p-coumaric acid (Myriam Richelle,: Isabelle Tavazzi,: and Elizabeth Afford, Comparison of the antioxidant activity of commonly consumed polyphenols beverages coffee cocoa and tea prepared per cup serving. Journal of Agriculture Food Chemistry 2001 49, 3438-3442).
  • Chlorogenic acid in green coffee has six sub groups namely, caffeoyl quinic acids (CQA), p-coumaroylquinic acids (PCQA), feruloylquinic acids (FQA), dicaffeoyl quinic acids (diCQA), caffeoylferuloyl quinic acids (CFQA) and feruloylcaffeoyl quinic acids (FCQA) (Clifford M.N chlorogenic acids In: coffee chemistry. Elsevier applied science, by Clarke RJ,Macrac (eds) 1985 London PP 153-202).
  • Extrazyme with declared activity of 7500 pectinase S Units and a multi enzyme complex containing a wide range of enzymes, including arabinase, cellulase, ⁇ -glucanase, hemi-cellulase and xylanase and (b) Energex in powder or liquid form, with declared activities of 50-120 fungal ⁇ -glucanase and 5000 to 12000 PSU per gram or ml, are used to facilitate the release of antioxidant constituents of coffee.
  • Coffee seeds are used as raw material for preparation of coffee conserve in WO02085397, whereas in present patent proposal coffee flakes are used as raw material for preparation of conserves, thereby overcoming the difficult operation of grinding the hard beans of green coffee.
  • the solvent in the referred patent is aqueous methanol, whereas in the present patent isopropanol and aqueous isopropanol, with an advantage of improved extractability for coffee constituents namely, phenolic compounds have been employed.
  • the main object of the present invention is to provide a process wherein coffee flakes are used as raw material for preparation of conserves, thereby overcoming the difficult operation of grinding the hard beans of green coffee.
  • Another object of the present invention is to provide a process for preparing coffee bean flakes for food purpose by using selected varieties of green coffee beans.
  • Yet another object of the present invention is to provide a process, wherein a higher yield of phenolic compounds (7.5%) is obtained.
  • a further object of the presentr invention is to provide a process wherein coffee flakes are treated with an enzyme mixture, which facilitates better release of coffee constituents namely phenolic compounds and chlorogenic acids for extraction purposes.
  • Still another object of the present invention is to provide a process wherein a mixture of aqueous solvents selected for higher efficiency in terms of extractability of coffee constituents is employed for extraction in place of conventional solvents.
  • the present invention provides an improved process for the preparation of antioxidant conserve from green coffee, which comprises, a) soaking green coffee beans in boiled water and/or optionally steaming the green coffee beans under pressure; b) flaking the coffee beans obtained in step (a), and drying the same till a predetermined level of moisture is obtained; c) roasting the dried flakes of step (b) and grinding the flakes to obtain flakes having size up to 15 mesh; d) drying the flakes as obtained in step (c) to a moisture level of 8 to 10 percent; e) de-fatting the dry coffee flakes as obtained in step (a) by conventional solvent extraction techniques; f) subjecting the de-fatted flakes as obtained in step (b) to enzyme treatment using the enzyme such as described herein within the enzyme to flakes ratio of 0.1 : 100 to 1.0 : 100, at a pH of 4.0 to 7.0 for a time period of 2 to 24 hours at a temperature ranging from 25 to 40 degree C, while maintaining the moisture level of flakes at 20 to 60 percent by
  • the coffee beans are soaked in water maintained at a temperature in the range of 50-100 0 C for a time period between 2 to 10 hours. In another embodiment of the present invention the coffee beans are steamed at a temperature in the range of 100 to 125 0 C for a period of 10 to 30 minutes at a pressure in the range of 1.0 to 1.5 kg/ cm 2 .
  • the thickness of flaked coffee bean is in the range of 0.5 to 5 mm.
  • drying is carried till a moisture level of
  • drying is carried out using a mechanical dryer or by a known method.
  • roasting is carried out at a temperature in the range of 150 to 200 0 C for a period of 1 to 10 minutes.
  • grinding is carried out to obtain flakes of size up to 10 mesh.
  • grinding is carried out using mechanical grinder.
  • the flakes so obtained have a shelf life up to twelve months.
  • the coffee bean flakes are coated with syrup having 20-100° Brix.
  • the coating of the syrup is carried out at a temperature in the range of 100 to 125 0 C for a period up to 30 minute.
  • the syrup used for coating is in the form of sugar or dextrin.
  • the coated flakes are dried, wherein drying is carried out for a period of 2 to 10 hours till moisture content is secured up to 8 to 10 %.
  • the drying of flakes are carried out using fluidized bed roaster at a specific temperature 190-195 0 C for a period of 1 to 1.5 minutes
  • the enzyme used is Energex, a proprietary preparation, with an activity of 50-120 units of fungal ⁇ -glucanase and 5000-12000 fungal pectinase S units per gm or ml, under conditions described as in the step (b) above.
  • the enzyme treated coffee flake powder is loaded into a series of extractors, up to four in number and extracted using a solvent mixture 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 preparing the conserve.
  • the coffee flakes after enzyme treatment are subjected to drying in a fluidized bed drier at 150-200 0 C for 1-5 minutes followed by drying at 50-55 0 C, to inactivate the enzymes and also bring down the moisture content to 8-10%.
  • the coffee conserve (200 -lOOOppm) obtained is used to improve the color stability of natural colorant preparation such as chilli or paprika based colorants for food use.
  • the coffee conserve is useful as a pharmaceutical and neutraceutical preparation with properties of quenching oxidative stress.
  • the process of the present invention consists of first selecting suitable green coffee beans such as robusta and arabica beans (cherry and parchument coffee) of various grades such as plantation A, B, AB and peaberry.
  • suitable green coffee beans such as robusta and arabica beans (cherry and parchument coffee) of various grades such as plantation A, B, AB and peaberry.
  • the green coffee beans are cleaned, and subjected to pretreatraent such as [a] deodorisation [b] removal of bitterness and [c] softening to obtain the deodorized softened coffee beans.
  • the deodorized softened green coffee beans are subjected to flaking operation to obtain 0.5 -5 mm thick flakes.
  • the flakes are passed through a suitable dryer preferably vibrofluidized bed roaster to get light cream or light brown colored crispy flakes. These flakes are subjected to a texturization process where by they become chewy.
  • These flakes are passed through a grinding mill to get grits of suitable size which are useful for the
  • the enzyme treated flakes is held at optimum temperature (37 0 C) for enzyme action.
  • suitable dryer preferably a vibrofluidized bed dryer to inactivate the enzymes and dry to 8-10% moisture to get light cream or light brown colored crispy flakes. This is followed by grinding to a coarse powder using a suitable grinder.
  • the flakes or powder is loaded in to columns and extracted with a binary solvent mixtures drawn from isopropanol, ethanol or methanol and water. It has been found that the selected binary solvent mixture are more efficient for the extraction of coffee constituents as compared to the individual solvents. This results in reduction of processing time and also lesser use of the solvents, besides improving recovery of the principal constituents, namely, phenolic compounds, which are mainly chlorogenic acids and diterpenes.
  • the extract or miscella is subjected to distillation under controlled conditions to maximize the recovery of the solvent and minimize the loss of constituents and to produce coffee conserve.
  • Example 1 is given by way of illustration and therefore should not be construed to limit the scope of the present invention.
  • Freshly harvested green coffee beans of arabica variety (lkg) obtained from a local orchard were subjected to steaming at 105 0 C (1.2 kg/cm 2 ) for 20 minutes.
  • the steamed warm green coffee beans were subjected to flaking to get 2 mm thick uniform flakes.
  • the flakes were passed through a vibro fluidized bed roster at 180 0 C for 40 seconds to get a crispy product free from bitterness.
  • the flakes (900 g) were allowed to cool before packing them in suitable containers.
  • the flakes can be used for various purposes. It is found that the shelf lives of the flakes are 11 month.
  • Freshly harvested green coffee beans of robusta variety (500 g) obtained from a local orchard was soaked in water maintained at an elevated temperature for 10 hours.
  • the soaked green coffee beans were flaked to get 3 mm thick uniform flakes.
  • the flakes were passed through a vibrofluidized bed roster at 200 0 C for 40 seconds to get a crispy product free from bitterness.
  • the flakes 450 g were texturized by addition hot sugar syrup at 100 0 C and holding for 30 minutes, followed by draining of the free syrup and drying of the flakes at 60 0 C to bring down the moisture to 8-10%.
  • the flakes (600 g) were allowed to cool before packing them in airtight containers. The flakes can be used for various purposes. It is found that the shelf life of the flakes is 12 months.
  • Coffee beans, pea berries of the arabica variety (lkg) obtained from a local orchard were subjected to steaming at 121 0 C for 20 minutes.
  • the steamed green coffee beans were flaked to get 2 mm thick uniform flakes.
  • the flakes were subjected to drying using cross flow dryer at a temperature of 55-60 0 C for suitable time period that is till light green colored crispy flakes are obtained.
  • the flakes are useful as a raw material for preparations of therapeutic value. It is found that the shelf life of the flakes is 10 months.
  • Table 1 depicts comparison of color values (Color was determined using Hunter color system), moisture of the control green coffee beans with the green coffee flakes (processed) during storage.
  • Table 2 depicts comparison of color values (Color was determined using Hunter color system), moisture of the control green coffee beans with the green coffee flakes (processed) of coffee flakes during storage.
  • Table 1 depicts comparison of color values (Color was determined using Hunter color system), moisture of the control green coffee beans with the green coffee flakes (processed) during storage.
  • Coffee flakes (10Og) obtained from arabica variety green beans were passed through a vibrofluidized bed dryer to get a crispy product free from bitterness.
  • the flakes were defatted with hexane by soxhlet extraction.
  • the solvent free/ dried flakes were passed through a hammer mill fitted with 30 mesh sieve to get a powder which was used as the raw material for preparation of conserves.
  • the powder (10Og) was loaded in a glass column and extracted with a solvent mixture containing 60 parts of methanol, and 40 parts of water, by volume. After the solvent addition a contact time of 2 hours was given and extract was drained out while simultaneously adding fresh solvent mixture on to the material in the column to keep it soaked for 1 hour.
  • the arabica green coffee flakes (10Og) obtained from green coffee beans was passed through a vibrofluidized bed dryer to get a crispy product free from bitterness.
  • the flakes were defatted with hexane by soxhlet apparatus.
  • a multi enzyme preparation was added as a suspension in water, at the concentration of 0.5% and pH 4.0, to defatted coffee flakes (20Og batch) and incubated at 37 0 C for 12 hours.
  • the control sample, that is 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 100° C for 30 minutes and 50 0 C using a conventional dryer to bring down the moisture level to about 10%.
  • the dry flakes were milled in a hammer mill fitted with 30-mesh (500 micron) to get a powder which was used as the raw material for preparation of conserves.
  • the treated and control samples lOOg each, were loaded in to separate glass columns and extracted with selected solvents, at a material to solvent ratio of 1:10.
  • the following solvent mixtures namely, isopropanol plus water (60:40) and ethyl alcohol plus water (60:40) were used for extraction.
  • Recoveries of coffee constituents with reference to yield of conserve, caffeine and chlorogenic acids were computed and comparative profile is presented in Table 3.
  • the chlorogenic acids in the conserve and yield of the conserve were higher in enzyme treated samples in comparison to control sample.
  • the antoxidant activity of the conserves was estimated by the ⁇ -carotene linoleic acid assay and DPPH model system and the results are included in Table 3.
  • the robusta green coffee flakes (20Og) were prepared from green coffee beans.
  • the flakes were extracted with ethyl acetate using soxhlet apparatus.
  • a multi enzyme preparation was added as a suspension in water, at the concentration of 0.5% and pH 4.0, to solvent free coffee flakes (20Og batch) and incubated at 37 0 C for 12 hours.
  • the control sample that is, without enzyme addition was also maintained under similar conditions of moisture (30%), pH and temperature. After the incubation period, both samples were charged to a vibrofiudized drier and held at 100 0 C for 30 minutes and subsequently at 50° C to bring down the moisture level to about 10%.
  • the dry flakes were passed through a hammer mill fitted with 30 mesh (500 micron) to get a powder which was used as the raw material for preparation of conserves.
  • the treated and control samples lOOg each, were loaded in to separate glass columns and extracted with selected solvents, at a material to solvent ratio of 1:10.
  • the following solvent mixtures namely, isopropanol plus water (60:40) and ethyl alcohol plus water (60:40) were used for extraction.
  • Recoveries of coffee constituents with reference to yield of conserve, caffeine and chlorogenic acids were computed and comparative profile is presented in Table 3.
  • the chlorogenic acids in the conserve and yield of the conserve were higher in enzyme treated samples in comparison to control sample.
  • the antoxidant activity of the conserves was estimated by the ⁇ - carotene linoleic acid assay and DPPH model system and the results are included in Table 3.
  • Example 7 Coffee flakes (20Og) prepared from low grade (Blacks, Bits and Browns) coffee and extracted with ethyl acetate using soxhlet apparatus. A multi enzyme preparation was added as a suspension in water, at the concentration of 0.5% and pH 4.0, to solvent free coffee flakes (20Og batch) and incubated at 37 0 C for 12 hours. The control sample, that is without enzyme addition was also maintained under similar conditions of moisture (30%), pH and temperature. After the incubation period both the samples were charged into a vibrofludized bed drier, held at 100 0 C for 30 minutes followed by drying at 50° C to bring down the moisture level to about 10%.
  • the dry flakes were passed through a hammer mill fitted with 30 mesh (500 micron) to get a powder which was used as the raw material for preparation of conserves.
  • the treated and control samples, lOOg each were loaded in to separate glass columns and extracted with selected solvents, at a material to solvent ratio of 1:10.
  • the following solvent mixtures namely, isopropanol plus water (60:40) and ethyl alcohol plus water (60:40) were used for extraction.
  • Recoveries of coffee constituents with reference to yield of conserve, caffeine and chlorogenic acids were computed and comparative profile is presented in Table 3.
  • the chlorogenic acids in the conserve and yield of the conserve were higher in enzyme treated samples in comparison to control sample.
  • the antoxidant activity of the conserves was estimated by the ⁇ -carotene linoleic acid assay and DPPH model system and the results are presented in Table 3.
  • Coffee conserve prepared from green coffee as described in above examples was used for chilli color stabilization. Chilli color 600 g (l,60,000cv) taken in a beaker was warmed over water bath and then mixed thoroughly. The coffee conserves (robusta and arabica 20 mg and 50 mg each separately) were dissolved in 0.5 ml propylene glycol (PG), added to the warmed chill color (100 g each batch), mixed using the magnetic stirrer and packed in 2 ml vials (1 gm each). The vials were stored at 4 0 C, 27 0 C and 42 0 C.
  • PG propylene glycol
  • TBHQ 200ppm was dissolved in 0.5 ml PG and added to the warm chilli color, mixed well and packed in 2 ml vials and stored at the above conditions.
  • chilli color was packed in 2ml vials and stored at the above conditions.
  • the samples were withdrawn from the above storage conditions and analyzed for color value using the spectrophotometric method. The results are presented in Table 4. After 90 days of storage, control samples stored at elevated temperature (42 0 C) showed highest color fading (1,32,000 from starting value of 1,60,000 cv) and control samples stored at 4 0 C also showed color fading (1,53,000 cv).
  • Novelty i.
  • the pretreatment and conditioning of green coffee bean flakes are novel approaches for extraction of coffee conserve ii.
  • Coffee flakes are treated with an enzyme mixture, which facilitates better release of coffee constituents namely phenolic compounds and chlorogenic acids for extraction purposes.
  • a mixture of aqueous solvents selected for higher efficiency in terms of extractability of coffee constituents is employed for extraction in place of conventional solvents.
  • Flaking will increase the surface area of the bean, which helps in further processing like size reduction, extraction, texturization etc. 2.
  • the flakes can be used for various purposes with special reference to new category of green coffee based food and beverage items and nutraceuticals beneficial to health.
  • the raw material is new to be utilized in food purpose like chocolates, coatings.
  • the process provides conversion of green coffee beans to flakes which does not contain bitterness to render edible flakes or grits. 5.
  • the present process includes preconditioning and removal of bitterness by using high temperature short time (HTST) method. 6.
  • HTST high temperature short time
  • the process is amenable for scale up to meet commercial demand.

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Abstract

The present invention deals with an improved process for extraction of an antioxidant conserve from Green coffee flakes by subjecting the flakes to enzyme treatment, which facilitates the extraction process. The process is novel in the use of Coffee flakes as raw material and very significant economically.

Description

ANTIOXIDANT CONSERVE FROM GREEN COFFEE AND PROCESS FOR ITS PREPARATION
Field of the invention The present invention relates to an improved process for the preparation of antioxidant conserve from green coffee.
Background and prior art of the invention
Coffee is one of the important agricultural commodities and which is cultivated globally on 11.6 million hectare. The productions of the agricultural commodities have been reached approximately up to 63 lakhs tons. Brazil and Colombia are accounted for nearly 44% of the world production for the same. The coffee is grown in about 80 countries across the globe and out of them near about 51 countries is considered to be the major producers for the coffee. As far as it is concerned to India, it is an agro-based rural enterprise. This industry is a major source of direct employment for about 4, 00,000 people in the area of cultivation of agro based products. Apart from this, the industry is also providing indirect employment to many people in processing and trade industry or auxiliaries. It is a highly labor-intensive industry, employing an estimated of about 100 million people are working all over 60 developing countries.
Today, in India, coffee occupies an important position among the export commodities.
More particularly, in the plantation sector. Production of coffee has risen from 18,000 tons during 1950s to 2,30,000 tons in 1998-99. The Indian coffee industry is also heading for the highest ever crop by the year 2004-2005 with an estimated crop of 4,00,000 tons {Indian Coffee 2002). Coffee beans are used after roasting and powdering mainly for the preparation of coffee beverage and instant coffee.
The green coffee beans yield caffeine as a by product during the coffee decaffeination process. The other reported prior art are related to use of green coffee powder for analytical purposes i.e. the analysis of green coffee constituents such as caffeine, chlorogenic acids etc. However, it is known that, coffee is a good source of phenolic compounds, which are known for use as antioxidants, anti-tumor agents and also posses a host of other therapeutic properties. The phenolic acids in coffee are mainly quinic acids with different amount of caffeoyl groups attached to its different positions. The phenolic acids present in coffee are such as chlorogenic acid, caffeic acid, para- coumaric acid and eugenol which have been shown preventive activities to exert cancer in animal models. The chlorgenic acid, which is the main phenolic acid in coffee, is able to protect the gastric mucosa against irritation, and therefore, improves the digestibility of foods, beverages and medicaments. The improved digestibility is expressed through a much reduced systemic acid secretion, which has been found to be directly dependent on an increased level of chlorogenic acid content. Also, chlorogenic acid has a chemopreventive effect on rat stomach cancer (Shimizu et ah, 1999). In general the natural chlorogenic acid content of coffee is reduced by approximately 40 to 80% during roasting process (Cliford et al., 1998). Thus, the use of green coffee bean will result in a product, which preserves the phenolic compounds. In the present work, processing conditions have been standardized for production of shelf-stable coffee bean flakes, which is microbiologically sound and capable to utilize for various purposes. The inventors are the first one whom has been made an attempt to develop a novel method for the processing of green coffee as a raw material which is being used for a variety of food purposes.
Green coffee contains a large amount of polyphenols exemplified by chlorogenic acid, caffeic acid, ferulic acid and p-coumaric acid (Myriam Richelle,: Isabelle Tavazzi,: and Elizabeth Afford, Comparison of the antioxidant activity of commonly consumed polyphenols beverages coffee cocoa and tea prepared per cup serving. Journal of Agriculture Food Chemistry 2001 49, 3438-3442). Chlorogenic acid in green coffee has six sub groups namely, caffeoyl quinic acids (CQA), p-coumaroylquinic acids (PCQA), feruloylquinic acids (FQA), dicaffeoyl quinic acids (diCQA), caffeoylferuloyl quinic acids (CFQA) and feruloylcaffeoyl quinic acids (FCQA) (Clifford M.N chlorogenic acids In: coffee chemistry. Elsevier applied science, by Clarke RJ,Macrac (eds) 1985 London PP 153-202). In addition to CQA, β- coffeoyl, 1-tryptopha, β-coffeoyl, 1-tyrosine and traces of unesterified e-caffeic acid, ferulic acid and 3-e-p-coumaric acid have been reported (Clifford M.N; kellard-B Birch- GG. Characterization of coffeoylferuloyl quinic acids by simultaneous isomerisation and transesterifϊcation with tetramethylammonium hydroxide. Food Chemistry 1989, 34, 2 81-288). Degradation of chlorogenic acids during roasting of green coffee was reviewed. Chlorogenic acid largely undergoes hydrolysis to caffeic acid and quinic acid during roasting. Since quinic acid occurs at about 0.5% in roast coffee, the hydrolysed acids undergo further chemical changes. The antioxidant activity of this diverse group of compounds present in coffee were compared with those of tea and wine (Myriam Richelle et al, 2001J.
Reference may be made to a patent (WO02085397, Slaga Barin et al., 2002-10-31) on therapeutic preparation from green coffee bean, resulting in a coffee extract containing natural antioxidants (phenolic compounds), and diterpenes (having detoxification properties) and has greater bioavailability in comparison to other polyphenol extracts.
In the present patent proposal, two commercial enzymes, namely a) Extrazyme (with declared activity of 7500 pectinase S Units and a multi enzyme complex containing a wide range of enzymes, including arabinase, cellulase, β-glucanase, hemi-cellulase and xylanase and (b) Energex in powder or liquid form, with declared activities of 50-120 fungal β-glucanase and 5000 to 12000 PSU per gram or ml, are used to facilitate the release of antioxidant constituents of coffee.
The methods reported in the hitherto known literature, have certain disadvantages which are overcome by the present patent application:
• Coffee seeds are used as raw material for preparation of coffee conserve in WO02085397, whereas in present patent proposal coffee flakes are used as raw material for preparation of conserves, thereby overcoming the difficult operation of grinding the hard beans of green coffee.
• Enzyme addition to the coffee flakes is a new approach to facilitate release of the constituents
• The yields of phenolic compounds as per the referred patent is 6.5%, in the present invention, a higher yield of phenolic compounds (7.5%) is obtained
• The solvent in the referred patent is aqueous methanol, whereas in the present patent isopropanol and aqueous isopropanol, with an advantage of improved extractability for coffee constituents namely, phenolic compounds have been employed.
Objects of the invention
The main object of the present invention is to provide a process wherein coffee flakes are used as raw material for preparation of conserves, thereby overcoming the difficult operation of grinding the hard beans of green coffee. Another object of the present invention is to provide a process for preparing coffee bean flakes for food purpose by using selected varieties of green coffee beans.
Yet another object of the present invention is to provide a process, wherein a higher yield of phenolic compounds (7.5%) is obtained.
A further object of the presentr invention is to provide a process wherein coffee flakes are treated with an enzyme mixture, which facilitates better release of coffee constituents namely phenolic compounds and chlorogenic acids for extraction purposes.
Still another object of the present invention is to provide a process wherein a mixture of aqueous solvents selected for higher efficiency in terms of extractability of coffee constituents is employed for extraction in place of conventional solvents.
Summary of the invention
Accordingly, the present invention provides an improved process for the preparation of antioxidant conserve from green coffee, which comprises, a) soaking green coffee beans in boiled water and/or optionally steaming the green coffee beans under pressure; b) flaking the coffee beans obtained in step (a), and drying the same till a predetermined level of moisture is obtained; c) roasting the dried flakes of step (b) and grinding the flakes to obtain flakes having size up to 15 mesh; d) drying the flakes as obtained in step (c) to a moisture level of 8 to 10 percent; e) de-fatting the dry coffee flakes as obtained in step (a) by conventional solvent extraction techniques; f) subjecting the de-fatted flakes as obtained in step (b) to enzyme treatment using the enzyme such as described herein within the enzyme to flakes ratio of 0.1 : 100 to 1.0 : 100, at a pH of 4.0 to 7.0 for a time period of 2 to 24 hours at a temperature ranging from 25 to 40 degree C, while maintaining the moisture level of flakes at 20 to 60 percent by weight; g) subjecting the enzyme treated flakes as obtained in step (c) to heat treatment at 100 to 150 degree C for 30 minutes followed by drying at 50 to 55 degree to reduce the moisture content to 8 to 10 percent to obtain creamy colored or light brown colored flakes; h) powdering the flakes as obtained in step (d) to a particle size of 25 to 40 mesh; i) charging the powder as obtained in step (e) in an extraction column by soaking in a solvent to water ratio of 1 :9 to 9: 1 at a temperature of 25 to 50 degree C and upto a period of 2 hours; j) optionally repeating the step (f) to obtain extract; Ic) pooling the extracts as obtained in step (f) and (g) and subjecting to de- solventization to reduce the solvent level in resulting conserve to a level of 20 to 30 ppm..
In one of the embodiments of the present invention the coffee beans are soaked in water maintained at a temperature in the range of 50-1000C for a time period between 2 to 10 hours. In another embodiment of the present invention the coffee beans are steamed at a temperature in the range of 100 to 125 0C for a period of 10 to 30 minutes at a pressure in the range of 1.0 to 1.5 kg/ cm2.
In still another embodiment of the present invention the thickness of flaked coffee bean is in the range of 0.5 to 5 mm. In yet another embodiment of the present invention drying is carried till a moisture level of
8 to 12 % remained in the coffee beans.
In a further embodiment of the present invention drying is carried out using a mechanical dryer or by a known method.
In an embodiment of the present invention roasting is carried out at a temperature in the range of 150 to 200 0C for a period of 1 to 10 minutes.
In another embodiment of the present invention grinding is carried out to obtain flakes of size up to 10 mesh.
In yet another embodiment of the present invention grinding is carried out using mechanical grinder. In a further embodiment of the present invention the flakes so obtained have a shelf life up to twelve months.
In still another embodiment of the present invention the coffee bean flakes are coated with syrup having 20-100° Brix. In another embodiment of the present invention the coating of the syrup is carried out at a temperature in the range of 100 to 125 0C for a period up to 30 minute. In yet another embodiment of the present invention the syrup used for coating is in the form of sugar or dextrin. In a further embodiment of the present invention the coated flakes are dried, wherein drying is carried out for a period of 2 to 10 hours till moisture content is secured up to 8 to 10 %.
In still further embodiment of the present invention, the drying of flakes are carried out using fluidized bed roaster at a specific temperature 190-1950C for a period of 1 to 1.5 minutes
In an embodiment of the present invention, the enzyme used is Energex, a proprietary preparation, with an activity of 50-120 units of fungal β-glucanase and 5000-12000 fungal pectinase S units per gm or ml, under conditions described as in the step (b) above. In another embodiment of the invention, the enzyme treated coffee flake powder is loaded into a series of extractors, up to four in number and extracted using a solvent mixture 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 preparing the conserve. In yet another embodiment of the invention, the coffee flakes after enzyme treatment are subjected to drying in a fluidized bed drier at 150-2000C for 1-5 minutes followed by drying at 50-550C, to inactivate the enzymes and also bring down the moisture content to 8-10%.
In still another embodiment of the invention, the coffee conserve (200 -lOOOppm) obtained is used to improve the color stability of natural colorant preparation such as chilli or paprika based colorants for food use.
In a further embodiment of the present invention, the coffee conserve is useful as a pharmaceutical and neutraceutical preparation with properties of quenching oxidative stress.
Detailed Description of the Invention
The process of the present invention consists of first selecting suitable green coffee beans such as robusta and arabica beans (cherry and parchument coffee) of various grades such as plantation A, B, AB and peaberry. The green coffee beans are cleaned, and subjected to pretreatraent such as [a] deodorisation [b] removal of bitterness and [c] softening to obtain the deodorized softened coffee beans. The deodorized softened green coffee beans are subjected to flaking operation to obtain 0.5 -5 mm thick flakes. The flakes are passed through a suitable dryer preferably vibrofluidized bed roaster to get light cream or light brown colored crispy flakes. These flakes are subjected to a texturization process where by they become chewy. These flakes are passed through a grinding mill to get grits of suitable size which are useful for the preparation of various food products. The flakes are also useful as raw material for the preparations of neutraceutical / therapeutic value using appropriate extraction and enrichment methods.
This is followed by treating the flakes with an aqueous solution of an enzyme mixture designed to cause cell wall degradation, and consisting of constituent enzymes namely, cellulase, hemicellulase, amylase, pectinase, arabinase, β-glucanase and xylanase. The enzyme treated flakes is held at optimum temperature (370C) for enzyme action. The flakes are passed through suitable dryer preferably a vibrofluidized bed dryer to inactivate the enzymes and dry to 8-10% moisture to get light cream or light brown colored crispy flakes. This is followed by grinding to a coarse powder using a suitable grinder. The flakes or powder is loaded in to columns and extracted with a binary solvent mixtures drawn from isopropanol, ethanol or methanol and water. It has been found that the selected binary solvent mixture are more efficient for the extraction of coffee constituents as compared to the individual solvents. This results in reduction of processing time and also lesser use of the solvents, besides improving recovery of the principal constituents, namely, phenolic compounds, which are mainly chlorogenic acids and diterpenes. The extract or miscella is subjected to distillation under controlled conditions to maximize the recovery of the solvent and minimize the loss of constituents and to produce coffee conserve.
The following examples are given by way of illustration and therefore should not be construed to limit the scope of the present invention. Example 1
Freshly harvested green coffee beans of arabica variety (lkg) obtained from a local orchard were subjected to steaming at 1050C (1.2 kg/cm2) for 20 minutes. The steamed warm green coffee beans were subjected to flaking to get 2 mm thick uniform flakes. The flakes were passed through a vibro fluidized bed roster at 1800C for 40 seconds to get a crispy product free from bitterness. The flakes (900 g) were allowed to cool before packing them in suitable containers. The flakes can be used for various purposes. It is found that the shelf lives of the flakes are 11 month.
Example 2
Freshly harvested green coffee beans of robusta variety (500 g) obtained from a local orchard was soaked in water maintained at an elevated temperature for 10 hours. The soaked green coffee beans were flaked to get 3 mm thick uniform flakes. The flakes were passed through a vibrofluidized bed roster at 200 0C for 40 seconds to get a crispy product free from bitterness. The flakes (450 g) were texturized by addition hot sugar syrup at 1000C and holding for 30 minutes, followed by draining of the free syrup and drying of the flakes at 60 0C to bring down the moisture to 8-10%. The flakes (600 g) were allowed to cool before packing them in airtight containers. The flakes can be used for various purposes. It is found that the shelf life of the flakes is 12 months.
Example 3
Coffee beans, pea berries of the arabica variety (lkg) obtained from a local orchard were subjected to steaming at 1210C for 20 minutes. The steamed green coffee beans were flaked to get 2 mm thick uniform flakes. The flakes were subjected to drying using cross flow dryer at a temperature of 55-600C for suitable time period that is till light green colored crispy flakes are obtained. The flakes are useful as a raw material for preparations of therapeutic value. It is found that the shelf life of the flakes is 10 months. Table 1 depicts comparison of color values (Color was determined using Hunter color system), moisture of the control green coffee beans with the green coffee flakes (processed) during storage.
Figure imgf000010_0001
Table 2 depicts comparison of color values (Color was determined using Hunter color system), moisture of the control green coffee beans with the green coffee flakes (processed) of coffee flakes during storage.
Figure imgf000010_0002
BRIEF DESCRIPTION OF TABLES
Table 1 depicts comparison of color values (Color was determined using Hunter color system), moisture of the control green coffee beans with the green coffee flakes (processed) during storage.
Wherein: L = lightness, A = positive red, B = positive yellow, DE = color difference Comparison of color values (L, A, B and DE) of the control green coffee beans with the green coffee flakes (processed) reveal that there is no difference or loss of color during storage. Table 2 depicts color determination of green coffee seeds, flakes and creamy colored flakes.
Wherein: L = lightness, A = positive red; negative green, b = yellow, DE = total color difference It can be noted that in the present invention there is no difference or loss of color during process and also on storage, this is be indirectly attributable to retention of Phenolic compounds in the end product. Phenolic compounds present in coffee have been shown preventive activities to exert cancer in animal models, improve digestibility of foods, beverages and medicaments.
Example 4
Coffee flakes (10Og) obtained from arabica variety green beans were passed through a vibrofluidized bed dryer to get a crispy product free from bitterness. The flakes were defatted with hexane by soxhlet extraction. The solvent free/ dried flakes were passed through a hammer mill fitted with 30 mesh sieve to get a powder which was used as the raw material for preparation of conserves. The powder (10Og) was loaded in a glass column and extracted with a solvent mixture containing 60 parts of methanol, and 40 parts of water, by volume. After the solvent addition a contact time of 2 hours was given and extract was drained out while simultaneously adding fresh solvent mixture on to the material in the column to keep it soaked for 1 hour. Totally 10 more such extracts were taken after a contact time of 1 hour every time. The extracts were pooled (1000 ml) and desolventised in a rota evaporator at atmospheric pressure till nearly 900 ml of the solvent was recovered. Rest of the distillation was carried out under reduced pressure of 20inches of mercury and a temperature of less than 500C 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 inches of mercury and a temperature below 800C for a period of 15 minutes by which time the solvent was fully removed from the product (2Og). The chlorogenic acid content in this conserve was 6.0% as expressed on raw material basis. The antioxidant activity of the conserves was estimated by the β-carotene linoleic acid assay and DPPH model system and the results are presented in Table 1. Example 5
The arabica green coffee flakes (10Og) obtained from green coffee beans was passed through a vibrofluidized bed dryer to get a crispy product free from bitterness. The flakes were defatted with hexane by soxhlet apparatus. A multi enzyme preparation was added as a suspension in water, at the concentration of 0.5% and pH 4.0, to defatted coffee flakes (20Og batch) and incubated at 370C for 12 hours. The control sample, that is 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 100° C for 30 minutes and 500C using a conventional dryer to bring down the moisture level to about 10%. The dry flakes were milled in a hammer mill fitted with 30-mesh (500 micron) to get a powder which was used as the raw material for preparation of conserves. The treated and control samples, lOOg each, were loaded in to separate glass columns and extracted with selected solvents, at a material to solvent ratio of 1:10. The following solvent mixtures namely, isopropanol plus water (60:40) and ethyl alcohol plus water (60:40) were used for extraction. Recoveries of coffee constituents with reference to yield of conserve, caffeine and chlorogenic acids were computed and comparative profile is presented in Table 3. The chlorogenic acids in the conserve and yield of the conserve were higher in enzyme treated samples in comparison to control sample. The antoxidant activity of the conserves was estimated by the β-carotene linoleic acid assay and DPPH model system and the results are included in Table 3.
Example 6
The robusta green coffee flakes (20Og) were prepared from green coffee beans. The flakes were extracted with ethyl acetate using soxhlet apparatus. A multi enzyme preparation was added as a suspension in water, at the concentration of 0.5% and pH 4.0, to solvent free coffee flakes (20Og batch) and incubated at 370C for 12 hours. The control sample that is, without enzyme addition was also maintained under similar conditions of moisture (30%), pH and temperature. After the incubation period, both samples were charged to a vibrofiudized drier and held at 1000C for 30 minutes and subsequently at 50° C to bring down the moisture level to about 10%. The dry flakes were passed through a hammer mill fitted with 30 mesh (500 micron) to get a powder which was used as the raw material for preparation of conserves. The treated and control samples, lOOg each, were loaded in to separate glass columns and extracted with selected solvents, at a material to solvent ratio of 1:10. The following solvent mixtures namely, isopropanol plus water (60:40) and ethyl alcohol plus water (60:40) were used for extraction. Recoveries of coffee constituents with reference to yield of conserve, caffeine and chlorogenic acids were computed and comparative profile is presented in Table 3. The chlorogenic acids in the conserve and yield of the conserve were higher in enzyme treated samples in comparison to control sample. The antoxidant activity of the conserves was estimated by the β- carotene linoleic acid assay and DPPH model system and the results are included in Table 3.
Example 7 Coffee flakes (20Og) prepared from low grade (Blacks, Bits and Browns) coffee and extracted with ethyl acetate using soxhlet apparatus. A multi enzyme preparation was added as a suspension in water, at the concentration of 0.5% and pH 4.0, to solvent free coffee flakes (20Og batch) and incubated at 370C for 12 hours. The control sample, that is without enzyme addition was also maintained under similar conditions of moisture (30%), pH and temperature. After the incubation period both the samples were charged into a vibrofludized bed drier, held at 1000C for 30 minutes followed by drying at 50° C to bring down the moisture level to about 10%. The dry flakes were passed through a hammer mill fitted with 30 mesh (500 micron) to get a powder which was used as the raw material for preparation of conserves. The treated and control samples, lOOg each, were loaded in to separate glass columns and extracted with selected solvents, at a material to solvent ratio of 1:10. The following solvent mixtures namely, isopropanol plus water (60:40) and ethyl alcohol plus water (60:40) were used for extraction. Recoveries of coffee constituents with reference to yield of conserve, caffeine and chlorogenic acids were computed and comparative profile is presented in Table 3. The chlorogenic acids in the conserve and yield of the conserve were higher in enzyme treated samples in comparison to control sample. The antoxidant activity of the conserves was estimated by the β-carotene linoleic acid assay and DPPH model system and the results are presented in Table 3.
Figure imgf000014_0001
Figure imgf000015_0001
Example 8
Coffee conserve prepared from green coffee as described in above examples was used for chilli color stabilization. Chilli color 600 g (l,60,000cv) taken in a beaker was warmed over water bath and then mixed thoroughly. The coffee conserves (robusta and arabica 20 mg and 50 mg each separately) were dissolved in 0.5 ml propylene glycol (PG), added to the warmed chill color (100 g each batch), mixed using the magnetic stirrer and packed in 2 ml vials (1 gm each). The vials were stored at 40C, 270C and 420C. TBHQ (200ppm) was dissolved in 0.5 ml PG and added to the warm chilli color, mixed well and packed in 2 ml vials and stored at the above conditions. For control, chilli color was packed in 2ml vials and stored at the above conditions. Once in thirty days, the samples were withdrawn from the above storage conditions and analyzed for color value using the spectrophotometric method. The results are presented in Table 4. After 90 days of storage, control samples stored at elevated temperature (420C) showed highest color fading (1,32,000 from starting value of 1,60,000 cv) and control samples stored at 40C also showed color fading (1,53,000 cv). Addition of TBHQ (200 ppm) and coffee conserves (500 ppm) to chilli color, gave protection against color fading, during storage at 4°C, 27°C and also at elevated temperature. The maximum protection against color fading was given by addition of robusta conserve (500 ppm) followed by arabica conserve (500ppm).
Table4: Chilli color stabilization using coffee conserves*
Figure imgf000015_0002
Figure imgf000016_0001
• Figures in the parenthesis indicate % of colour loss over starting value of cv. 1,60,000
• CR: Conserve prepared from robusta coffee • CA: Conserve prepared from arabica coffee
Novelty: i. The pretreatment and conditioning of green coffee bean flakes are novel approaches for extraction of coffee conserve ii. Coffee flakes are treated with an enzyme mixture, which facilitates better release of coffee constituents namely phenolic compounds and chlorogenic acids for extraction purposes.
Ui. A mixture of aqueous solvents selected for higher efficiency in terms of extractability of coffee constituents is employed for extraction in place of conventional solvents.
ADVANTAGES OF THE INVENTION:
1. Flaking will increase the surface area of the bean, which helps in further processing like size reduction, extraction, texturization etc. 2. The flakes can be used for various purposes with special reference to new category of green coffee based food and beverage items and nutraceuticals beneficial to health.
3. The raw material is new to be utilized in food purpose like chocolates, coatings.
4. The process provides conversion of green coffee beans to flakes which does not contain bitterness to render edible flakes or grits. 5. The present process includes preconditioning and removal of bitterness by using high temperature short time (HTST) method. 6. The process is amenable for scale up to meet commercial demand.

Claims

We claim:
1. An improved process for the preparation of an antioxidant conserve from green coffee flakes, which comprises: a) soaking green coffee beans in boiled water and/or optionally steaming the green coffee beans under pressure; b) flaking the coffee beans obtained in step (a), and drying the same till a predetermined level of moisture is obtained; c) roasting the dried flakes of step (b) and grinding the flakes to obtain flakes having size up to 15 mesh; d) drying the flakes as obtained in step (c) to a moisture level of 8 to 10 percent; e) de-fatting the dry coffee flakes as obtained in step (a) by conventional solvent extraction techniques; f) subjecting the de-fatted flakes as obtained in step (b) to enzyme treatment using the enzyme such as described herein within the enzyme to flakes ratio of 0.1 : 100 to 1.0 : 100, at a pH of 4.0 to 7.0 for a time period of 2 to 24 hours at a temperature ranging from 25 to 40 degree C, while maintaining the moisture level of flakes at 20 to 60 percent by weight; g) subjecting the enzyme treated flakes as obtained in step (c) to heat treatment at 100 to 150 degree C for 30 minutes followed by drying at 50 to 55 degree to reduce the moisture content to 8 to 10 percent to obtain creamy colored or light brown colored flakes; h) powdering the flakes as obtained in step (d) to a particle size of 25 to 40 mesh; i) charging the powder as obtained in step (e) in an extraction column by soaking in a solvent to water ratio of 1:9 to 9:1 at a temperature of 25 to 50 degree C and upto a period of 2 hours, j) optionally repeating the step (f) to obtain extract; k) pooling the extracts as obtained in step (f) and (g) and subjecting to de- solventization to reduce the solvent level in resulting conserve to a level of 20 to 30 ppm.
2. A process as claimed in claim 1, wherein in step (a) the coffee beans are soaked in water maintained at a temperature in the range of 50 to 100 0C for a time period between 2 to 10 hours.
3. A process as claimed in claim 1, wherein in step (a) the coffee beans are steamed at a temperature in the range of 100 to 125 0C for a period of 10 to 30 minutes at a pressure in the range of 1.0 to 1.5 kg/ cm2.
4. A process as claimed in claim 1, wherein in step (b) thickness of flaked coffee bean is in the range of 0.5 to 5 mm.
5. A process as claimed in claim 1, wherein in step (b) drying is carried out up to a moisture level 8 to 12 % remained in the coffee beans.
6. A process as claimed in claim 1, wherein in step (c) roasting is carried out at a temperature in the range of 150 to 200 0C for a period of 1 to 10 minute.
7. A process as claimed in claim 1, wherein step (c) grinding is carried out using mechanical grinder.
8. A process as claimed in claim 1, wherein the coffee bean flakes as obtained in step
(c) are coated with syrup having 20-100° Brix.
9. A process as claimed in claim 1, wherein the drying is preferably done by using any conventional dryer such as a cross flow or hot air dryer or a fluidized bed dryer.
10. A process as claimed in claim 1, wherein the solvent used for de-fatting the flakes is preferably selected from hexane or petroleum.
11. A process as claimed in claim 1, wherein the enzyme used mainly consists of constituent enzymes namely cellulase, hemicellulase, amylase, pectinase, arabinase, β-glucanase and xylanase.
12. A process as claimed in claim 1, wherein the solvent used to charge the coffee flakes is preferably selected from methanol and isopropanol.
13. A process as claimed In claim 1 wherein the solvent to water ratio is preferably 6:4 to 7:3 and the solvent is preferably isopropanol.
14. Coffee bean flakes as obtained by the process as claimed in claim 1.
15. An antioxidant conserve from green coffee flakes obtained by the process as claimed in claiml, comprising chlorogenic acids - 60%; caffeic acid - 10%; ferulic acid - 16%; cumeric acid - 10%; unidentified fraction - 4%, wherein the conserve exhibits 65 to 73% antioxidant activity.
16. An improved process for the preparation of an antioxidant conserve from green coffee flakes substantially as herein described with reference to the foregoing examples.
PCT/IB2006/000681 2005-03-31 2006-03-27 Antioxidant conserve from green coffee and process for its preparation WO2006103515A1 (en)

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EP1844661A1 (en) * 2006-04-10 2007-10-17 Kraft Foods Holdings, Inc. Stabilzed enzyme compositions
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