WO2007062206A2

WO2007062206A2 - Chocolate formulations with functional properties

Info

Publication number: WO2007062206A2
Application number: PCT/US2006/045376
Authority: WO
Inventors: Michael Z. Martin
Original assignee: Mid-South Technologies
Priority date: 2005-11-23
Filing date: 2006-11-24
Publication date: 2007-05-31
Also published as: WO2007062206A3

Abstract

This invention relates to methods of concentrating bioactive substances from cacao (Theobroma spp.) seeds. The invention further relates to methods of processing, and concentrating various classes of chemical compounds from foodstuffs so that a full spectrum food concentrate containing all of the major compound classes present in the raw foodstuff is extracted retaining the full nutritional or bioactive value of the starting material. The invention also relates to formulations, pharmaceutical preparations, nutritional preparations, and dietary supplements that may be prepared with the extracted materials and their uses to treat various ailments or nutritional deficiencies.

Description

Chocolate Formulations with Functional Properties

Background

1. Field of the Invention

This invention is directed to bioactive substances from cacao and methods for concentrating extractions. The invention is also directed to formulations, pharmaceuticals preparations, nutritional preparations, and dietary supplements extracted from food materials.

2. Description of the Background

The seeds of Theobroma cacao L. and other Theobroma species have been used by Mesoamerican cultures as a source of a stimulating and health-giving beverage since before recorded history. The use of cacao in these cultures was so important that at the time of the invasion of the Aztec empire by the Spaniard Cortez, cacao seeds were used as currency.

The original beverage as made by the indigenous cultures combined the whole ground cacao seeds with various other botanicals including capsicum, vanilla pods, the flowers of Magnolia, Cymbopetalum, Quararibea funebris, etc. These hot infusions were generally taken unsweetened (sugar cane being unknown in the New World at that time) although honey was produced and available. The Spanish took back the cacao seeds to Europe, where milk and sugar were added to suit European tastes, and modern chocolate was born.

Over time, the refining and manufacture of chocolate evolved into a science. The main goals of refining were the reduction of bitterness through treatment with mild alkali (the "dutching" process developed in the Netherlands) and increasing the "silky" mouthfeel of solid chocolate through the addition of additional milkfat, cocoa butter and/or other seed fats in a process of "conching" or prolonged folding of the semi-liquid mass. These goals served to achieve a product that was, and remains, very acceptable to consumers. However, these processes inherently served to greatly reduce the healthy properties of the raw cocoa seed.

It has been discovered that both the conching and dutching processes oxidize the phenolic compounds in cocoa, thereby reducing the antioxidant and other beneficial properties. Furthermore the large amounts of cane sugar and milkfat, cocoa butter and/or other seed fats greatly contribute to the caloric content of the chocolate. There is clearly a need for healthier chocolate products. It is the object of the present invention to provide these healthier products, since the methods of the present invention yield products chemically identical to the phytochemical profile of the cacao seed, with all the nutritional qualities intact.

The conching process previously mentioned has as its primary goal the creation of a smooth, creamy mouthfeel. This is accomplished by the mechanical reduction of particle size from 100 microns or greater in diameter to approximately 20 microns. The current art of conching achieves this reduction through extensive refolding of the semi- liquid mass which process aerates and slowly mechanically reduces the particle size in the mass. While each manufacturer has their own recipe, conching times of from 10 to 14 hours and as long as two days are known in the art. In order to keep the cocoa mass in a semi-liquid state, the mass is kept heated during the conching process. For a greater understanding of the traditional chocolate manufacturing process, refer to, for example, "Cocoa and Chocolate - Health and Nutrition" Ian Knight, edit.; "Chocolate, Cocoa and Confectionary Science and Technology 3rd Edition" Bernard Minifle; and "Industrial Chocolate Manufacture and Use" S.D. Beckett edit, which are herein incorporated by reference.

The prolonged heating and aeration which occurs during the conching process is a major cause of oxidizing and degrading the native phenolic compounds found in the raw cocoa seed, with the resultant loss of their health-giving properties. One method of the present invention is to replace conching with the use of mechanical shearing to reduce particle size. It has been discovered that a smooth mouthfeel can be achieved through the use of suitable high-shear mixing equipment. This elimination of the conching step has surprisingly been found to greatly reduce the oxidation of phenolic and other health- promoting compounds in the cocoa mass, while at the same time retaining the mouthful characteristics associated with a quality chocolate product. A further advantage is the reduction of time required to achieve a final product, since mechanical shearing is accomplished in a matter of seconds or minutes, rather than hours or days.

An example of a suboptimum extraction procedure is provided in U.S. No. 5,897,866 issued to Bombardelli et al. on April 27, 1999. This document teaches that of the a) aromatic hydrocarbons, b) aliphatic hydrocarbons, and c) halohydrocarbons, hexane and methylene chloride work best for extraction of lycopene from tomato, particularly with a fatty acid additive. This patent describes a multi-step process where lycopene is removed from tomato material by homogenizing, heating and cooling, and centrifugation or filtering, prior to an extraction step with the selected solvent. Alternatively tomato skins are sieved, treated in water for an hour, dried and ground prior to extraction with a hexane solvent. Following extraction and evaporation, a 5% lycopene material was obtained, in a yield of approximately 0.5 gm per 10 kg of starting material. To obtain a more purified material from the lycopene oil, supercritical carbon dioxide was used in a countercurrent three step column, yielding 24 gms of pure product from 500 gm of starting oil. However, a variety of tomatoes contain approximately 100 mg per 100 gm weight (0.1%) ("Lycopene content in Raw Tomato Varieties and Tomato Products by EMAN M. TAWFIK (paper presented at IFT Annual Meeting on the Technical Program Session, June 18, 2002, Anaheim, CA. USA). Thus the extraction procedure taught in the Bombardelli patent recovers only about 1 percent of the lycopene available and the majority of the lycopene is wasted. Obviously other procedures are needed to commercialize lycopene production from this source.

Similar pretreatment procedures are used for other categories of bioactive substances such as the solvent extraction and chromatography purification of anthocyanins, shown in U.S. 6,423,365 issued July 23, 2002 to Nair. In each case, plant cellular material generally first must be disrupted, a solvent is added to remove the bioactive substance, and then the removed substance is further purified. As mentioned in this patent, unless a very crude material is desired that contains sugars and other undesirable components, an overly expensive step is required to prepare the bioactive substance.

A further problem that has not been addressed sufficiently by this field is the inability to simultaneously extract multiple desired bioactive components from a given plant material. For example, many fruits have desirable quantities of lycopenes and anthocyanins, tannins, condensed tannins (proanthocyanins) and other substances. ' In order to meet the demand for these materials as neutraceuticals, high efficiency systems are needed that can capture each of these bioactive agents as desired from each plant material. The crude extraction and partial purification procedures known thus far lack both the ability to obtain a suitably wide spectrum of nutrients as well as control the ratios of bioactive compounds, as needed for specific desired effects. Materials with more controllable bioactive compositions and methods for their economical preparation are needed to advance this field and human nutrition generally.

Summary

The present invention overcomes the problems and disadvantages associated with current strategies and designs and provides new tools and methods for providing chocolate with nutritional value.

One embodiment of the invention is directed to a method of placing raw cacao nib, partially or completely fermented cacao nib, or combinations thereof in a suitable container and extracting the material with water, organic solvents, near-critical or supercritical fluids or combinations thereof in order to achieve a complete recovery of all representative compound classes present in the material.

Another embodiment of the invention is directed to a method of combining the CSC with a product made from the fruit of certain palm trees, in particular the acai palm (Euterpe oleraceά)..

Another embodiment of the invention is directed to a method of combining the CSC with sterols and/or sterolins to provide a product with cholesterol-lowering, immune system enhancing, or other beneficial health properties. Other embodiments and advantages of the invention are set forth in part in the description, which follows, and in part, may be obvious from this description, or may be learned from the practice of the invention.

Description of the Invention

As embodied and broadly described herein, the present invention is directed to formulations and methods for extraction of bioactive substances from cacao seeds.

Methods for extracting these bioactive compounds generally favor one type of bioactive molecule over another. This unfortunately diminishes the value or raises the cost of the product supplied to the consumer. Many methods in the literature for isolating bioactive molecules or groups of molecules originated from basic research on small samples and are not practical for commercial use. Commercial value reflects consumer desires for large quantities of bioactive substances at a reasonable cost. Presently, neutraceuticals are available in mostly rough form and obtained by crude extractions of plant materials that are particularly rich in a desired bioactive compound or group of compounds. Accordingly, the consumer often purchases a product having a composition that is determined by the ease of a particular extraction procedure or availability of a plant having a high concentration of particular compound. Most desired, in contrast, are products that fully exploit the range of bioactivities available in nature, and that comprise different classes of substances that share common effects.

A method of the present invention which comprises placing raw cacao nib (i.e. broken seeds), partially or completely fermented cacao nib, or combinations thereof in a suitable container and extracting the material with water, organic solvents, near-critical or supercritical fluids or combinations thereof in order to achieve a complete recovery of all representative compound classes present in the material. It may be necessary to extract the material with any or all of the extractants in order to adequately remove the various compound classes present. Additionally, it may be necessary to sequentially extract the same material with a series of different extractants in order to elute desirable constituents with each succeeding extractant. Furthermore, it may be necessary to vary the conditions of each phase of extraction in order to optimize the recovery of certain constituents. It may also be necessary to treat the material to be extracted at some point in the process with enzymes to aid in the release of desired compounds. The end result is a cacao seed concentrate (CSC) containing the full spectrum of biologically active compounds present in the cacao seed.

Another embodiment comprises combining the CSC with similarly prepared concentrates of other foodstuffs that exhibit nutritional or health-giving properties.

Another embodiment comprises combining the CSC with a product made from the fruit of certain palm trees, in particular the acai palm {Euterpe oleracea). It has surprisingly been found that the fruit of this palm has a flavor profile that is very harmonious with that of chocolate and cacao seed products, as it is a non-acid fruit. The acai fruit contains high levels of anthocyanins and other phytochemicals that have been shown to possess beneficial nutritional properties. These phytochemicals being phenolic in nature, have been found to blend well flavorwise with the CSC whose compounds are likewise predominantly phenolic. An advantage of this addition of acai phytochemicals to the CSC is that the antioxidant potential of the blend is increased over that of the CSC alone. This results in a chocolate product with improved nutritional and/or health-giving properties.

Another embodiment comprises combining the CSC with a fungal material possessing biologically active compounds.

Another embodiment comprises fungal material is a mycelium, or a version of the mycelium that has been concentrated by various means including but not limited to the methods of the present invention.

Another embodiment comprises the combination of fungal material and CSC is designed to contain fungal polysaccharides. These polysaccharides are known to stimulate the immune system. It has been found that the combination also reduces or eliminates the need for the addition of mono- and disaccharides such as cane sugar to the CSC in order to achieve a palatable product. The result is a chocolate product with improved nutritional and/or health-giving properties Another embodiment comprises the CSC is combined with sterols and/or sterolins to provide a product with cholesterol-lowering, immune system enhancing, or other beneficial health properties. It has been found that sterols derived from certain kinds of biomass with the methods of the present invention, when combined with CSC₅ possess a harmonious flavor blend as well as enhancing the overall chocolate taste profile.

Another embodiment comprises the CSC is combined with biologically-active carbohydrates to provide a product with beneficial health properties. ,

Another embodiment comprises the CSC is combined with carotenoids or carotenoid-containing oleoresins to provide a product that has increased antioxidant or other beneficial health properties.

Another embodiment comprises the CSC is combined with poly-unsaturated fatty acids.

Another embodiment comprises the CSC is combined with herbs or medicinal plants, their extracts or concentrates.

Another embodiment comprises the CSC is combined with a concentrate of a foodstuff as outlined in U.S. Patent Application No. 10/660,737 "Full Range Nutritional Supplements and Methods for Their Manufacture" and related PCT and national filings, herein incorporated by reference.

In an embodiment, whole, partly degraded, ground or crushed cacao is pumped into the sealable chamber, optionally contacted with a co-solvent and then contacted with a solvent in the liquid phase so as to charge the solvent with analyte. Charged solvent is collected and removed to isolate the analyte. In an embodiment, the cacao material contacts the solvent after sealing the chamber and air has been removed. The resulting mixture of solvent and natural source is maintained under pressure so that the natural source and solvent are in intimate contact to charge the solvent with analyte. This type of extraction may be carried out in any vessel that can be sealed and evacuated of air as required. The extraction may be performed at any suitable temperature and is preferably carried out at or below room temperature. The extracting fluid or gas preferably is introduced at one side or location of the plant raw material column and passes through the column, solvating and picking up phytochemicals that enter the fluid as it traverses the column. The extracted fluid leaves the column space and enters another space where the fluid material is removed, leaving extracted phytochemicals behind as for example described in WO0072861 published 12/07/00 for ASHRAF-KHORASSANI MEHDI et al. The pressure used within the cacao column depends on the type of solvent/gas and the type of phytochemical(s) to be extracted. Preferably the extraction material comprises carbon dioxide, molecular nitrogen (nitrogen gas), hydrogen, an aliphatic or halide carbon compound such as butane, propane, freon, or a mixture such as carbon dioxide with an alcohol, carbon dioxide with ethanol, carbon dioxide with methanol, carbon dioxide with 15% ethanol, and carbon dioxide with alcohol and with isopropyl amine as a secondary modifier. One discovery is that supercritical fluid may be used to remove a wider spectrum of phytochemicals.

Yet another discovery is that a sub-critical pressure may be used to obtain a wide spectrum extract without the higher cost and hazard associated with higher supercritical pressures. Such "sub-critical" pressures generally are between 0.05 and 0.95 times the supercritical pressure for a given temperature, preferably are between 0.25 to 0.8 times and more preferably between 0.5 and 0.7 times the supercritical pressure. For example, the complex tannins from many types of plant materials often are incompletely eluted at low pressures with a single water or water based solvent (water plus water-miscible organic solvent such as an alcohol, or at a high or low pH). It was found that sub critical conditions with carbon dioxide at a pressure between 0.5 and 0.67 times the supercritical pressure for a given temperature often removes more of this group of phytochemicals.

In many cases a single supercritical fluid may be used such as carbon dioxide, propane, butane, isobutane and the like. It was found through experimentation that addition of a small amount of secondary solvent often yields improved extraction. For example, addition of an alcohol such as methanol or ethanol or ethyl acetate as a cosolvent to, for example a concentration of about 0.02% to 10% (mole ratio) and preferably between 0.1% to 5% of a carbon dioxide solvent can improve recovery of phytochemical.

In another discovery solvents used either at sub-critical pressures or super critical pressures were found to extract material with functional properties from cacao. Samples such as whole berries, tomato skins, fruit processing waste, lightly minced herb matter and the like may be treated by the pressure step without any previous processing step. This advantageous feature greatly lowers cost and increases convenience of processing cacao into extracts. Thus, waste streams may be used directly for low cost high volume extractions.

Another procedure that was discovered and which improves economies of scale, is the direct addition of a co-solvent to the raw material at any time prior to sub critical or super critical chromatography. It was found that a co-solvent such as methanol, ethanol or ethyl acetate may be added at a typical ratio of about 0.1% to 25% of the weight of the sample, and preferably between 0.3% to 5% of the weight of the sample prior to addition of the high pressure solvent. Optionally the sample can be treated with a vacuum after adding the co-solvent and before adding the high pressure solvent.

Another discovery was that nitrogen gas can be used in sub critical conditions with a co-solvent for high efficiency extractions. The co-solvent may be added to the raw material prior to application of high pressure. The co-solvent also may be introduced at the same time as or after the addition of high pressure nitrogen, hi one embodiment an alcohol such as methanol, ethanol, propanol or butanol is added and subjected to the high pressure nitrogen, and flowed through the raw material chromatography space. In yet another embodiment the secondary solvent is introduced at an inlet at a separate location and passes through the raw material chromatography space at the high pressure. In another embodiment the secondary solvent is added prior to or after exposing the raw material to a vacuum. The nitrogen gas generally is pressurized to between 100 to 2000 psi, preferably between 300 to 1500 psi, more preferably between 500 to 1200 psi and yet more preferably between 700 to 900 psi. Without wishing to be bound by any one theory for how this embodiment of the invention operates, it is believed that high pressure nitrogen increases the activity of the co-solvent and thereby decreases the amount of co- solvent needed, which lowers solvent costs and improves extraction efficiency.

Low Pressure Aqueous Phase Raw Material Chromatography

Of course, high pressure extraction may be combined with low pressure aqueous extraction method(s). In one embodiment an aqueous phase such as water with up to 40% ethanol or methanol and optionally at up to 60 degrees centigrade is passed through the raw material chromatography space to remove water solutes such as flavonoides. This is followed by high pressure extraction as described above. High pressure extraction under weak solvating conditions (lower pressure and/or temperature) such as liquid carbon dioxide at subcritical conditions removes for example polar compounds such as fatty acids and sterols. If most of the bioactive substances are such compounds then the high pressure extraction preferably occurs without an aqueous extraction step. Organic, less polar substances such as polycyclics preferably can be removed by using conditions that are closer to supercritical, or by switching to supercritical conditions. Accordingly, if different classes of substances need to be removed separately, an aqueous phase extraction may be followed by not only one but two high pressure extractions. For a full spectrum extract at lowest cost, however, it is desirable to subject the original plant material to a simple high pressure extraction.

Even though high pressure raw material chromatography is very good at removing a wide range of substances, it was found that a pre-extraction with low pressure aqueous phase can actually improve recovery from a subsequent high pressure step.

Representative Extraction Agents

A wide variety of solvents appropriate for solvating various bioactive substances including, but not limited to, alcohols, weak acids, ketones, chloro derivatives, hydrocarbons, fluorinated hydrocarbons, acetates, ethers, or a combination thereof. Due to it's non-flammable nature, as opposed to propane or butane, and excellent solvating properties for a wide range of target analytes, CO₂ has become the most common volatile substance used in the art of supercritical fluid extraction, and is desirable for many embodiments. However, CO₂ in the presence of water can form carbonic acid, which can degrade biomolecules and some metal surfaces used for reaction vessals. Additionally, supercritical CO₂ extraction systems often operate at temperatures in excess of 39°C. Holding labile natural materials at such temperatures for long periods during processing may result in thermally or enzymatically induced spoilage. On the other hand significant dissolution was found using sub-critical carbon dioxide conditions as described herein. Non-chlorinated fluorocarbon solvents also can be used, both at sub-critical concentrations and in supercritical conditions. Such solvents as represented by the disclosure of U. S. Pat. No. 5,512,285 are useful for embodiments.

In one embodiment, non-chlorinated fluorocarbon solvents including, but not limited to, trifluoromethane, difluoromethane, fluoromethane, pentafluoroethane, 1, 1, 1,- trifluoroethane, 1, 1-difluoroethane, 1,1,1,2,2,3,3-heptafluoropropane, 1,1,1,3,3,3- hexafluoropropane, 1,1,1,2,2-pentafluoropropane, 2,2,3 -hexafluoropropane, 1,1,2,2,3,3- hexafluoropropane, 1,1,1,2,3,3, hexafluoropropane, and 1,1,1,2-tetrafluoroethane may be used. A mixture of these solvents may be used to tailor the boiling point of the mixture to a particular process and facilitate the selective elution of specific bioactive substances. The solvent may be further modified by mixing with another volatile substance such as butane, hexane, ethanol or any other appropriate substance. In a preferred embodiment, the non-fluorocarbon solvent used for extraction is a tetrafluoroethane, preferably 1,1,1,2- tetrafluoroethane. In a further preferred embodiment, the tetrafluoroethane is unmodified.

Other embodiments and uses of the invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. AU references cited herein, including all publications, U.S. and foreign patents and patent applications, are specifically and entirely incorporated by reference. It is intended that the specification and examples be considered exemplary only with the true scope and spirit of the invention indicated by the following claims.

H

Claims

1. A method for extracting bioacti ve substances comprising: placing raw cacao nib in a suitable container; extracting the raw cacao nib material with water, organic solvents, near-critical or supercritical fluids or combinations thereof in order to achieve a complete recovery of all representative compound classes present in the material.

2. The method of claim 1 , wherein the extracted cacao is combined with a product made from the fruit of certain palm trees, in particular the acai palm {Euterpe oleraced).

3. The method of claim 1, wherein the extracted cacao is combined with sterols and/or sterolins to provide a product with cholesterol-lowering, immune system enhancing, or other beneficial health properties.