DIETARY SUPPLEMENT DERIVED FROM NATURAL PRODUCTS BY HOT MELT EXTRUSION (HME) PROCESSING

Abstract

The invention provides a composition rich in flavonoids based on natural extracts, comprising a flavonoid extract dispersed by melt mixing or extrusion and encapsulated in a polymer matrix. The invention is also a novel dietary supplement with the naturally occurring ingredient ()-epicatechin, cacao extracted, that could potentially prevent or reduce the risk of Atherosclerotic pathology. The use of ()-epicatechin is promising as a therapeutic agent due to its potential antioxidant activity and its diverse biological properties. ()-Epicatechins are chemically unstable and extensively degraded in fluids of near neutral or greater pH, such as intestinal juice and bile. Current technologies used for taste masking and modified release as spray drying, liposome entrapment, co-crystallization, freeze drying, among others, suffer from numerous shortcomings, including poor repeatability and limitations on target delivery.

Claims

1. A composition rich in flavonoids based on natural extracts, comprising a flavonoid extract dispersed by melt mixing or extrusion and encapsulated in a polymer matrix.

2. The composition of claim 1, wherein said extract is at least one from the group consisting of cocoa extract, cacao extract, tea extract, Vitis vinifera seeds and skin extract, Persea americana leaves, peel and seed extract, Allium cepa extract, Allium sativum extract, Vaccinium oxycoccos and Vaccinium macrocarpon extract, Nasturtium officinale extract, Petroselinum crispum extract, Vitis vinifera fruit and red wine extract, Citrus reticulate, Citrus sinensis, Citrus limon and Citrus paradise, Olea europaea leaves and fruits extract, Garnicia mangostana extract, Garcinia species extract selected from the group consisting of G. cambogia, G. kola, G. madruno, and mixtures thereof.

3. The composition of claim 1, wherein said polymer matrix is selected from the group consisting of poly (acrylic acid), poly (ethylene oxide), poly (ethylene glycol), poly (vinyl pyrrolidone), poly (vinyl alcohol), polyacrylamide, poly (isopropyl acrylamide), poly (cyclopropyl methacrylamide), ethyl cellulose, carboxymethyl cellulose, hydroxyethyl cellulose, propyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, cellulose acetate phthalate, alginic acid, carrageenan, chitosan, hyaluronic acid, pectinic acid, (lactide-co-glycolide) polymers, starch, sodium starch glycolate, polyurethane, silicones, polycarbonate, polychloroprene, polyisobutylene, polycyanoacrylate, poly (vinyl acetate), polystyrene, polypropylene, poly (vinyl chloride), polyethylene, poly (methyl methacrylate), poly (hydroxyethyl methacrylate), acrylic acid, butyl acrylate copolymer, 2-ethylhexyl acrylate and butyl acrylate copolymer, vinyl acetate and methyl acrylate copolymer, ethylene vinyl acetate and polyethylene terephthalate, ethylene/vinyl acetate copolymer and polyethylene, polyethylene terephthalate, cellulose, methyl cellulose, hypromellose acetate succinate nf, hypromellose acetate succinate jp, hypromellose acetate succinate, hypromellose phthalate nf, hypromellose phthalate, low-substituted hydroxypropyl cellulose nf, low-substituted hydroxypropyl cellulose jp, low-substituted hydroxypropyl cellulose nflow-substituted hydroxypropyl cellulose jp copolymer, hypromellose usp, hypromellose ep, hypromellose jp, hypromellose phthalate jp, hypromellose phthalate ep, hypromellose, hypromellose phthalate nf, hypromellose phthalate, low-substituted hydroxypropyl cellulose, methacrylates, cellulose acetate butyrate, polylactide-polyglycolide copolymers, polycaprolactone, polylactide, polyglycolide, polyvinylpyrrolidone-co-vinyl acetate, polyrethanes, polyvinyl caprolactampolyvinyl acetatepolyethylene glicol graft copolymer, polyvinyl caprolactam, polyvinyl acetate, vinylpyrrolidonevinyl acetate copolymer, vinyl-pyrrolidone polymer, polyvinylacetate, polyoxyethylenepolyoxypropylene copolymer, polyoxy-ethylene, polyoxypropylene, polyoxirane, povidone, polyethylene oxide, cellulose acetate, copovidone, povidone K12, povidone K17, povidone K25, povidone K30, povidone K90, hypromellose E5, hypromellose E4m, hypromellose K3, hypromellose K100, hypromellose K4m, hypromellose K100m, hypromellose phthalate HP-55, hypromellose phthalate HP-50, hypromellose acetate succinate 1 grade, hypromellose acetate succinate m grade, hypromellose acetate succinate h grade, cellulose acetate phthalate, cationic methacrylate, methacrylic acid copolymer type A, methacrylic acid copolymer type B, methacrylic acid copolymer type C, polymethylacrylates, polyvinyl alcohol, hydroxypropylmethylcellulose acetate succinate, ethyl acrylate, methyl methacrylate, trimethylammonioethyl methacrylate, ethyl acrylate-methyl methacrylate copolymer, butyl/methyl methacrylate-dimethylaminoethyl methacrylate copolymer, butyl/methyl methacrylate, dimethylaminoethyl methacrylate, methacrylic acid-ethyl acrylate copolymer, methacrylic acid, methacrylic acid-methyl methacrylate copolymer, methyl acrylate-methyl methacrylate-methacrylic acid copolymer, methyl acrylate, methyl methacrylate and diethylaminoethyl methacrylate copolymer, methyl methacrylate, diethylaminoethyl methacrylate, succinate, d--tocopheryl polyethylene glicol 100 succinate, d--tocopheryl polyethylene glicol, ethylene oxide, polypropylene oxide, polyvinyl alcohol-polyethylene glycol graft copolymer, methacrylic acid-ethyl acrylate copolymer, poloxamer, micronized poloxamer, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, ethylene glycol-vinyl alcohol graft copolymer, polydextrose nf, hydrogenated polydextrose nf, methacrylic acid copolymer, methacrylic acid and methyl methacrylate copolymer, methacrylic acid and ethyl acrylate copolymer, carbomer homopolymer, carbomer copolymer, carbomer interpolymer and mixtures thereof.

4. The composition of claim 1, which exhibits taste masking characteristics up to 120 seconds in saliva in the absence of flavoring or sweeteners or related agents.

5. The composition of claim 1, which exhibits taste masking characteristics and successive modified release of minimum 80% up to 240 minutes, wherein the first 120 minutes occurs in a medium of pH=1.2, and the rest 120 minutes in a medium of pH=6.8.

6. The composition of claim 1, having modified release characteristics and having a particle-size distribution between 250 m and 425 m.

7. The composition of claim 1, having modified release characteristics and having a particle-size distribution between 180 m and 250 m.

8. The composition of claim 1, having modified release characteristics and having a particle-size distribution between 125 m and 180 m

9. The composition of claim 1, having modified release characteristics and having a particle-size distribution up to 125 m

10. The composition of claim 1, having modified release characteristics in the absence of coatings, crosslinking or chemical reactions affecting said composition.

11. A method for manufacturing the composition of claim 1, comprising the steps of: (a) melt mixing or extruding the extract under operating conditions and screw configuration to protect the characteristics of said natural extract or mixtures thereof; and (b) milling the resulting composition to a powder under operating conditions suitable to protect the characteristics of the natural extract or mixtures thereof.

12. The method of claim 11, further including the step of adding the resulting milled powder to a food or a matrix.

13. A composition manufactured according to claim 11, wherein said composition is used in a dietary supplement, food, functional food, nutraceuticals and pharmaceuticals.

14. A composition manufactured according to claim 11, wherein said composition has thermal stability, light and moisture resistance.

15. A taste masking composition rich in flavonoids based on natural extracts, comprising a flavonoid extract dispersed by melt mixing or extrusion and encapsulated in a polymer matrix.

16. The composition of claim 15, wherein said extract is at least one from the group consisting of cocoa extract, cacao extract, tea extract, Vitis vinifera seeds and skin extract, Persea americana leaves, peel and seed extract, Allium cepa extract, Allium sativum extract, Vaccinium oxycoccos and Vaccinium macrocarpon extract, Nasturtium Officinale extract, Petroselinum crispum extract, Vitis vinifera fruit and red wine extract, Citrus reticulate, Citrus sinensis, Citrus limon and Citrus paradise, Olea europaea leaves and fruits extract, Garnicia mangostana extract, Garcinia species extract selected from the group consisting of G. cambogia, G. kola, G. madruno, and mixtures thereof.

17. The composition of claim 15, wherein said polymer matrix is selected from the group consisting of poly (acrylic acid), poly (ethylene oxide), poly (ethylene glycol), poly (vinyl pyrrolidone), poly (vinyl alcohol), polyacrylamide, poly (isopropyl acrylamide), poly (cyclopropyl methacrylamide), ethyl cellulose, carboxymethyl cellulose, hydroxyethyl cellulose, propyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, cellulose acetate phthalate, alginic acid, carrageenan, chitosan, hyaluronic acid, pectinic acid, (lactide-co-glycolide) polymers, starch, sodium starch glycolate, polyurethane, silicones, polycarbonate, polychloroprene, polyisobutylene, polycyanoacrylate, poly (vinyl acetate), polystyrene, polypropylene, poly (vinyl chloride), polyethylene, poly (methyl methacrylate), poly (hydroxyethyl methacrylate), acrylic acid, butyl acrylate copolymer, 2-ethylhexyl acrylate and butyl acrylate copolymer, vinyl acetate and methyl acrylate copolymer, ethylene vinyl acetate and polyethylene terephthalate, ethylene/vinyl acetate copolymer and polyethylene, polyethylene terephthalate, cellulose, methyl cellulose, hypromellose acetate succinate nf, hypromellose acetate succinate jp, hypromellose acetate succinate, hypromellose phthalate nf, hypromellose phthalate, low-substituted hydroxypropyl cellulose nf, low-substituted hydroxypropyl cellulose jp, low-substituted hydroxypropyl cellulose nflow-substituted hydroxypropyl cellulose jp copolymer, hypromellose usp, hypromellose ep, hypromellose jp, hypromellose phthalate jp, hypromellose phthalate ep, hypromellose, hypromellose phthalate nf, hypromellose phthalate, low-substituted hydroxypropyl cellulose, methacrylates, cellulose acetate butyrate, polylactide-polyglycolide copolymers, polycaprolactone, polylactide, polyglycolide, polyvinylpyrrolidone-co-vinyl acetate, polyrethanes, polyvinyl caprolactampolyvinyl acetatepolyethylene glicol graft copolymer, polyvinyl caprolactam, polyvinyl acetate, vinylpyrrolidonevinyl acetate copolymer, vinyl-pyrrolidone polymer, polyvinylacetate, polyoxyethylenepolyoxypropylene copolymer, polyoxy-ethylene, polyoxypropylene, polyoxirane, povidone, polyethylene oxide, cellulose acetate, copovidone, povidone K12, povidone K17, povidone K25, povidone K30, povidone K90, hypromellose E5, hypromellose E4m, hypromellose K3, hypromellose K100, hypromellose K4m, hypromellose K100m, hypromellose phthalate HP-55, hypromellose phthalate HP-50, hypromellose acetate succinate 1 grade, hypromellose acetate succinate m grade, hypromellose acetate succinate h grade, cellulose acetate phthalate, cationic methacrylate, methacrylic acid copolymer type A, methacrylic acid copolymer type B, methacrylic acid copolymer type C, polymethylacrylates, polyvinyl alcohol, hydroxypropylmethylcellulose acetate succinate, ethyl acrylate, methyl methacrylate, trimethylammonioethyl methacrylate, ethyl acrylate-methyl methacrylate copolymer, butyl/methyl methacrylate-dimethylaminoethyl methacrylate copolymer, butyl/methyl methacrylate, dimethylaminoethyl methacrylate, methacrylic acid-ethyl acrylate copolymer, methacrylic acid, methacrylic acid-methyl methacrylate copolymer, methyl acrylate-methyl methacrylate-methacrylic acid copolymer, methyl acrylate, methyl methacrylate and diethylaminoethyl methacrylate copolymer, methyl methacrylate, diethylaminoethyl methacrylate, succinate, d--tocopheryl polyethylene glicol 100 succinate, d--tocopheryl polyethylene glicol, ethylene oxide, polypropylene oxide, polyvinyl alcohol-polyethylene glycol graft copolymer, methacrylic acid-ethyl acrylate copolymer, poloxamer, micronized poloxamer, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, ethylene glycol-vinyl alcohol graft copolymer, polydextrose nf, hydrogenated polydextrose nf, methacrylic acid copolymer, methacrylic acid and methyl methacrylate copolymer, methacrylic acid and ethyl acrylate copolymer, carbomer homopolymer, carbomer copolymer, carbomer interpolymer and mixtures thereof.

18. The composition of claim 15, having modified release characteristics and having a particle-size distribution between 250 m and 425 m.

19. The composition of claim 15, having modified release characteristics and having a particle-size distribution between 180 m and 250 m.

20. The composition of claim 15, having modified release characteristics and having a particle-size distribution between 125 m and 180 m.

21. The composition of claim 15, having modified release characteristics and having a particle-size distribution up to 125 m

Description

BRIEF DESCRIPTION OF THE FIGURES

[0038] FIG. 1 shows the chemical structure of flavanols and procyanidin (DP5).

[0039] FIG. 2 illustrated the chromatograpic profile by HPLC for the reference material.

[0040] FIG. 3 describes the first heat Cacao Extract Differential Scanning calorimetry (DSC) characterization.

[0041] FIG. 4 shows the Cacao extract Differential Scanning calorimetry (DSC) Cooling curve. characterization.

[0042] FIG. 5 illustrates the second heat thermal analysis Cacao extract Differential Scanning calorimetry (DSC) characterization.

[0043] FIG. 6 describes the cacao extract Thermogravimetric analysis (TGA) characterization.

[0044] FIG. 7 features the cacao extract Isothermal Thermogravimetric characterization.

[0045] FIG. 8 illustrates the Cacao extract Oxygen Induction Time (OIT) characterizationStability Analysis.

[0046] FIG. 9 describes the Cacao extract Fourier-Transform Infrared Spectrocopy (FTIR) characterization.

[0047] FIG. 10 shows the first heating thermal analysis of ()-Epicatechin at 90% Differential Scanning calorimetry (DSC) characterization.

[0048] FIG. 11 features another first heating thermal analysis Cocoa Extract Differential Scanning calorimetry (DSC) characterization.

[0049] FIG. 12 illustrates the release profile of polyphenols in artificial saliva at 23 C. for examples I to V.

[0050] FIG. 13 describes the Release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. for examples I to V.

[0051] FIG. 14 shows the Oxygen Induction Time (OIT) for stability analysis of encapsulated formulations vs cacao extract for examples I to V.

[0052] FIG. 15 shows the release profile of polyphenols in artificial saliva at 23 C. for example VI.

[0053] FIG. 16 describes the Thermogravimetric analysis of cacao extract vs an encapsulated formulation from example VII.

[0054] FIG. 17 illustrates Release profile of polyphenols in artificial saliva at 23 C. for examples VIII and IX.

[0055] FIG. 18 shows the release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. for examples VIII and IX.

[0056] FIG. 19 describes the torque and melt temperature behavior of examples X to XIII FIG. 20 describes the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for examples X to XIII

[0057] FIG. 21 illustrates the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for examples X to XIII

[0058] FIG. 22 features release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. examples X to XIII FIG. 23 illustrates the torque and melt temperature behavior of examples XIV to XVII.

[0059] FIG. 24 shows the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for examples X to XIII at given particle size distribution.

[0060] FIG. 25 features the release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. for examples X to XIII FIG. 26 illustrates the torque and melt temperature behavior of example XVIII.

[0061] FIG. 27 shows the thermal characterization of example XVIII.

[0062] FIG. 28 describes the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for example XVIII.

[0063] FIG. 29 features the release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. for example XVIII.

[0064] FIG. 30 describes the torque and melt temperature behavior of example XIX.

[0065] FIG. 31 illustrates the thermal characterization of example XIX.

[0066] FIG. 32 shows the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for example XIX.

[0067] FIG. 33 describes the release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. for example XIX.

[0068] FIG. 34 illustrates the torque and melt temperature behavior of example XX.

[0069] FIG. 35 features the thermal characterization of example XX.

[0070] FIG. 36 illustrates the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for example XX.

[0071] FIG. 37 describes the release profile of epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. for example XX.

[0072] FIG. 38 shows the torque and melt temperature behavior of example XXI.

[0073] FIG. 39 describes the thermal characterization of example XXI.

[0074] FIG. 40 illustrates the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for example XXI.

[0075] FIG. 41 shows the release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. for example XXI.

[0076] FIG. 42 is a comparative radar graphic for taste profile in chocolate candy evaluated (examples XXII).

[0077] FIG. 43 is comparative radar graphic for taste profile in cereal bars evaluated (examples XXIII, XIV, XV, XVI, XVII).

[0078] FIGS. 44 and 45 show two possible screw configurations useful for melt extrusion.

SUMMARY OF THE INVENTION

[0079] The invention is directed to a composition rich in flavonoids based on natural extracts, comprising a flavonoid extract dispersed by melt mixing or extrusion and encapsulated in a polymer matrix.

[0080] The invention also relates to a taste masking composition rich in flavonoids based on natural extracts, comprising a flavonoid extract dispersed by melt mixing or extrusion and encapsulated in a polymer matrix.

[0081] The invention is a novel dietary supplement with the naturally occurring ingredient ()-epicatechin, cacao extracted, that could potentially prevent or reduce the risk of Atherosclerotic pathology. The use of ()-epicatechin is promising as a therapeutic agent due to its potential antioxidant activity and its diverse biological properties. ()-Epicatechins are chemically unstable and extensively degraded in fluids of near neutral or greater pH, such as intestinal juice and bile. Current technologies used for taste masking and modified release as spray drying, liposome entrapment, co-crystallization, freeze drying, among others, suffer from numerous shortcomings, including poor repeatability and limitations on target delivery.

[0082] In order to modify the rate of release and protect epicatechins from degradation in the gastrointestinal tract, polymeric systems such as polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer, polyvinyl acetate-povidone copolymer, methacrylic acid-methyl methacrylate copolymer, ethylcellulose, among others, could be used as carriers. This would provide benefits such as protection from rapid degradation, modified release and prolonged duration of bioactive agents.

[0083] The instant invention provides a product made by hot melt extrusion for oral consumption containing a bioactive compound such as epicatechin, cacao extract. Furthermore, nutraceutical formulations rich in flavonoids or other sort of antioxidants have been developed by using Hot Melt Extrusion (HME) or continuous melt mixing techniques. The corresponding compound consist of about 30-60% wt. of natural extract and polymers GRAS type (Generally Regarded as Safe) as taste masking and release agents.

[0084] The formulations have to be extruded at a temperature substantially below the melting point of the interest molecules which guaranties that a significant degradation of these molecules does not occur. In order to monitor any possible chemical and thermal change on the formulations, the samples ought to be characterized by using different techniques, such as simple and Oxidative Induction Times Testing (OIT), Differential Scanning calorimetry (DSC), Thermogravimetric analysis (TGA); likewise, chemical evaluations may be carried out by using chromatographic and spectrophotometric techniques.

[0085] The invention also provides a food ingredient derived from cacao, with high content of procyanidins (epicathechin, catechin and other flavanols), by HME processing for taste masking. The invention further provides a dietary supplement for human use, using epichatechin as an active pharmaceutical ingredient, by HME processing.

[0086] The invention provides nutraceutical formulations by using Hot Melt Extrusion (HME) which may be scaled and commercially launched either as a nutraceutical or pharmaceutical products or functional food ingredients.

[0087] The present invention is also directed to a novel dietary supplement or food additive with the naturally occurring ingredient ()-epicatechin and catechins extracted from cacao, that could potentially prevent or reduce the risk of the atherosclerotic pathology. The structures of cathechins are of the family having structures such as:

##STR00001##

DETAILED DESCRIPTION OF THE INVENTION

[0088] The pharmaceutical industry is facing two main problems, having poorly soluble drugs that require an increased dosage formulation so the proper drug absorption can be guaranteed, and the low bioavailability of the drug due to deficient dissolution during its passage through the gastrointestinal tract. Different approaches can be applied to overcome the solubility and bioavailability problems. One of them is manufacturing of solid dispersions; systems where one component, such as an API, is dispersed in a carrier, usually polymeric, and where the whole system appears to be in a solid state.

[0089] There are different types of solid dispersions, but only 3 can be achieved by HME, crystalline solid dispersion, amorphous solid dispersion, and solid solutions. Crystalline solid dispersions are systems wherein the crystalline drug substance is dispersed into an amorphous carrier matrix. The Differential Scanning calorimetry (DSC) profile for such a system is characterized by the presence of a melting endotherm (Tm) corresponding to the crystalline API and a characteristic glass transition temperature (Tg) corresponding to the amorphous carrier. They are generally designed to achieve controlled drug release profiles for highly soluble drugs.

[0090] Amorphous solid dispersions result when a melt extruded drug-polymer mixture is cooled at a rate that does not allow the drug to recrystallize or processed at temperatures at which the drug melts but remains immiscible with the carrier. The DSC profile for this system is characterized by the presence of two Tg. They have a potential to revert to the more stable crystalline form. In a solid solution, the drug molecule is molecularly dissolved in the polymeric carrier matrix and exhibits a single Tg. An amorphous solid solution is a pharmaceutically desirable single-phase system preferably including an amorphous polymer as the carrier, a drug in its high-energy state, as well as other excipients such as processing aids, recrystallization inhibitors, and wetting agents. A better understanding of the structure of a solid dispersion, particularly the existing physical form of a drug in the polymer excipient is necessary to predict the stability, solubility and hence bioavailability of melt extrudates.

[0091] Hot-Melt Extrusion (HME) is a recognized process that has been used in the last two decades for the manufacturing of solid dispersions. It has become very popular in the pharmaceutical field because it is a continuous process, solvent free, easy to clean and can be used for the preparation of different drug delivery systems; including granules, pellets, sustained released tablets, suppositories, stents, ophthalmic inserts, and transdermal and transmucosal delivery systems. Since it is a continuous process, fewer steps are involved resulting in lower cost of production.

[0092] HME is a process where a material that melts or softens under elevated temperatures and pressures is forced through an orifice by screws to produce polymeric products of uniform shape and density. It is carried out using an extruder, a barrel containing one or two rotating screws that transport material down the barrel. Optimization of process parameters, characterization and performance evaluation of the product, and assessment of its stability are inevitable tasks for successful application of HME in pharmaceutical formulations. The solid dispersion can be analyzed by different techniques such as Differential Scanning calorimeter (DSC),

[0093] Thermogravimetric Analysis (TGA), rheometry, X-Ray Diffraction (XRD), and microscopy, among others. One of the challenges of generating solid dispersions with HME is the tendency of the API to recrystallize after the temperature drops from elevated processing temperature to room temperature. Different strategies can be employed to address the recrystallization issue, for example, if the goal is to improve bioavailability of the drug via increasing the APIs dissolution rate, then choosing appropriate excipients and/or optimizing the HME process is needed to improve the drug-polymer miscibility or dramatically slow down the recrystallization rate.

[0094] For HME applications, the polymer excipient has to present thermoplastic characteristics, it must be thermally stable at the extrusion temperature employed, the Tg should be between 50 and 180 C., it should exhibit low hygroscopicity to avoid crystallization, and it has to be no toxic. Hot-stage microscopy (HSM), DSC and rheological analysis can be used to measure HME processing temperatures and design HME process and formulations. Low Tm and Tg from the polymeric excipient enable the low temperature extrusion process and makes the solubility analysis easier since the phase separation and recrystallization (dissolution kinetics) are faster when compared to polymeric excipients with high Tg. However, fast kinetics are not desirable if the API recrystallization is what should be avoided. Theoretically, Tg can be calculated with Fox equation (Eq. 1)

[00001]$\begin{array}{cc}\frac{1}{T}=\frac{w_1}{T_{g.Math..Math.1}}+\frac{w_2}{T_{g.Math..Math.2}}& \left(\mathrm{Eq}..Math.1\right)\end{array}$

if the sample is based on two components, where w1 and w2 refer to the weight fraction and Tg1 and Tg2 to the glass transition temperatures of drug and polymeric excipient, respectively. The Gordon Taylor equation has been applied as well to drug-polymer samples to study the miscibility of the binary components.

[0095] It has been shown that the dissolution behavior of HME solid dispersion depends on the physicochemical characteristics of the excipient(s) applied, therefore, the choice of excipients plays an important role in a successful formulation. Different polymers as excipients can be employed to prepare immediate and sustained release dosage forms via HME. Polyethylene oxide (PEO), polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), hydroxypropyl methyl cellulose (HPMC), hydroxypropyl cellulose (HPC), vinylpyrrolidone-vinylacetate copolymer (Kollidon VA 64), dimethylaminoethyl methacrylate copolymer (Eudragit E), PEG 6000-vinyl caprolactam-vinylacetate copolymer, and polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol graft co-polymer (Soluplus) can be used as immediate release (IR) polymeric excipients. On the other hand, ethylene vinyl acetate (EVA), polyvinyl acetate (PVA), polyL-lactic acid (PLA), polylactic-co-glycolic acid (PLGA), polycaprolactone, silicone, ammonium methacrylate copolymer (Eudragit RS/RL), polyvinyl acetate-polyvynilpyrrolidone (Kollidon SR), and lipid matrices (microcrystalline wax, stearic acid, carnauba wax, etc.) can be used as sustained release (SR) polymeric excipients.

[0096] The most suitable pair (API-excipient or API-excipient combination IR/SR) can improve the drug release profile, and samples that have a more sustained release because they are less porous and have better mechanical properties can be produced by HME.

[0097] Little is found in the literature concerning API solubility in a polymeric excipient after HME processing and storage at a given temperature. Some researchers have used HSM, DSC and rheological analysis to characterize acetaminophen dissolution in PEO. The samples were prepared with increasing loads of acetaminophen and by the HME process. FIG. 2 shows the different temperatures of acetaminophen dissolved in PEO; this diagram can be interpreted as a phase diagram. In region A acetaminophen and PEO form a liquid solution and are fully miscible, in region B acetaminophen does not totally dissolve and there are solid drug particles.

[0098] Therefore, it is more favorable to process acetaminophen-PEO formulations in region A. In region C (solid dispersion region) acetaminophen can molecularly disperse in PEO and it can partially recrystallize. This kind of phase diagram is very useful because valuable information can be obtained to formulate and develop the HME process; it is based on the API dissolution in the polymer excipient at different temperatures and increasing the API loading dose.

[0099] Hydrophobic excipients including polyvinylpyrrolidone and polyvinylpyrrolidone-co-vinyl acetate, polyethylene glycols, poly-ethylene oxides, some celluloses, polymethacrylate derivatives and a polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (Soluplus) have been used to enhance the solubility and bioavailability of poorly water soluble active ingredients using HME techniques.

[0100] The polymers showing their physical properties in Tables 1-4 are particularly suitable for the invention.

TABLE-US-00001 TABLE 1 Tm Tg Tdeg Density Polymer ( C.) ( C.) ( C.) (g/cm.sup.3) Extracto de cacao ~200 1.2 ()-Epicatechin 90% ~242 ~260 Aqualone EC-N7 123.75/164.32/196.70 ~255 1.14 Kollidon SR 42.56 ~220 1.2 Soluplus 77.29 ~270 1.082 Eudragit L100 81.24 ~250 11.887 Ethocel Standard 10 174.95 127.61 ~220 1.251 Klucel EF 192.03 ~247 1.296

TABLE-US-00002 TABLE 2 Commercial Tm Tg Tdeg Chemical name name ( C.) ( C.) ( C.) Polyvinylpyrrolidone Kollidon 12PF N.A. ~90 ~225 Vinylpyrrolidone-vinyl Kollidon VA 64 N.A. ~101 ~238 acetate copolymer Polyethylene glycol PEG 3350 ~53-57 Polyvinyl acetate - Kollidon SR 42.56 ~220 Polyvinylpyrrolidone Hydroxypropyl Aqoat AS-HG 57.1 135 218.8 Methylcellulose Acetate Succinate Methylcellulose Metolose 60SH ~163 ~280-300 Hydroxy propyl Klucel HF. GF, 100-150 >250 cellulose LF Coplymer of Plasdone S-630 109-112 >300 N-vinyl-2- pyrrolidone and vinyl acetate Ethylcellulose Aquaion N7, N22 N.A. ~156 >250 Modified Starch Starch 1500 ~300 ~300 Polylactic acid PLA ~155 ~65 ~250 Methacrylic Acid - Eudragit L100/ 81.24 ~250 Methyl Eudragit E PO Methacrylate Copolymer Ethylcellulose Ethocel standard 174.95 127.61 ~250 10

TABLE-US-00003 TABLE 3 Commercial Tm Tg Tdeg Chemical name name ( C.) ( C.) ( C.) Polyvinyl- Kollidon N.A. ~90 ~225 pyrrolidone 12PF vinylpyrrolidone- Kollidon N.A. ~101 ~238 vinyl acetate VA 64 copolymer Polyethylene glycol PEG 3350 ~53-57 Polyvinyl acetate- Kollidon 42.56 ~220 polyvinylpyrrolidone SR Hydroxypropyl Methylcellulose Aqoat 57.1 135 218.8 Acetate Succinate AS-HG Methylcellulose Metolose ~163 ~280-300 60SH Hydroxy propyl Klucel HF. 100-150 >250 cellulose GF, LF Coplymer of N- Plasdone 109-112 >300 vinyl-2-pyrrolidone S-630 and vinyl acetate Ethylcellulose Aqualon N.A. ~156 >250 N7, N22 Modified Starch Starch 1500 ~300 ~300 Polylactic acid PLA ~155 ~65 ~250 Methacrylic Eudragit 81.24 ~250 Acid-Methyl L100/ Methacrylate Eudragit Copolymer E PO Ethylcellulose Ethocel 174.95 127.61 ~250 standard 10

TABLE-US-00004 TABLE 4 BASF TG TM MW pH RESTRICTIONS KOLLIDON VA 64 101 45K independent 30 149 50K independent 90 156 1.25M independent CL cross-linked KOLLICOAT IR MAE 30 45 208 45K independent emulsion MAE 100 DP 250K >5.5 EVONIK Protect 45 47K <5.0 EUDRAGUAR Control emulsion Biotic emulsion

[0101] In its broadest aspect the invention provides the taste masking, characterization, stability and functionality of an encapsulated cacao extract, dissolution (including release) and other in-vitro tests required to demonstrate its efficiency.

[0102] The present invention provides a novel dietary supplement or food additive with the naturally occurring ingredient ()-epicatechin and catechins extracted from cacao, that are particularly useful to prevent or reduce the risk of the atherosclerotic pathology.

[0103] In carrying out the instant invention, compounds rich in polyphenols (incl. cacao extract) are polymer encapsulated for taste masking and modified release. The product of the invention rich in polyphenols (incl. cacao extract) has modified release independent of particle size between 120 m and 425 m.

[0104] Similarly, the compound rich in polyphenols (incl. cacao extract) are polymer encapsulated for thermal stability and moisture resistance. The product of the invention containing compounds rich in polyphenols (incl. cacao extract) do not include taste masking agents or processing aids or flavor additives or sweeteners. The formulations of the invention are made by continuous melt mixing process for manufacturing said compound rich in polyphenols with special screw configuration to guarantee dispersion and considering the low melting point of cacao extract (10-19 C.).

[0105] The product of the invention which includes compounds rich in polyphenols (incl. cacao extract) does not include coatings, crosslinking or chemical reactions affecting the said compound. The cacao extract of the invention is characterized as shown below.

Chemical Characterization of Cacao Extract

[0106] Flavanols and procyanidins are specific classes of flavonoids. Procyanidins are the oligomers of the monomeric flavanols (i.e., epicatechin and catechin) as shown in FIG. 1. The molecular weight of the flavanols oligomers is expressed as their degree of polymerization (DP). Separation based on DP permits the capture of the large structural diversity. Specifically, for cacao flavanols and procyanidins have been quantified up to a predefined molecular weight cut of DP=10 by summing oligomerig fractions DPI-DP10 (Robbins et al., 2012). Nevertheless, an additional method was used in order to quantify independently the monomers and xanthines. FIG. 1 illustrates the chemical structure of flavanols and procyanidin (DP5), modified from (Robbins et al., 2012).

[0107] The determination of favanol and procyanidin (by degree of polymerization 1-10) content of cacao flavanol extract was also conducted. The determination of flavanol and procyanidins content of cacao extract was based on a normalized AOAC method. This methodology is applicable to the determination of flavanols and procyanidins (DP1-DP10) content of chocolate, cacao liquors, cacao powders and cacao extracts. The sum of monomeric (DP=1) and oligomeric fractions (DP2-DP10) is reported as the total procyanidins content.

Sample Preparation

[0108] The cacao extract was extracted with hexane to remove their lipid content components prior to extraction of flavanols and procyanidins. Flavanols and procyanidins (DP1-DP10) were extracted with an acidified aqueous acetone solvent system [acetone: water: acetic acid (AWAA). Then, the extract was passed through SPE cartridges Strata SCX, 55 |im particle size and pore 70 A (Phenomenex 8B-S10-HBJ, California, Estados unidos), filtered and transferred to vials for normal-phase HPLC analysis. The method of calibration for this protocol was ()-epicatechin and the relative response factors (RRFs) for DP2-10 (Table 5).

TABLE-US-00005 TABLE 5 Relative response factors for fractions DP1-DP10 under HPLC conditions (Robbins et al., 2012) Oligomeric fraction Relative response factor DP1 (monomers) 1.0 DP2 (dimers) 0.374 DP3 (trimers) 0.331 DP4 (tetramers) 0.249 DP5 (pentamers) 0.237 DP6 (hexamers) 0.198 DP7 (heptamers) 0.169 DP8 (octamers) 0.139 DP9 (nonamers) 0.116 DP10 (decamers) 0.121

High Performance Liquid Chromatography-FLD Parameters

[0109] The identification, integration and quantification of the chromatographic signals were performed in a HPLC with fluorescence detector (FLD) (Agilent 1200). The column used was a Develosil Diol 100 A 2504.6 mm, 5 p,m particle size (Phenomenex, Torrance, Calif.). Temperature of the oven was kept at 35 C. and the flow rate was 1 mL/min. The injection volume was 5 |iL. The mobile phase consisted of acidic acetonitrile [(A) CH.sub.3CNHOAc, 98+2 (v/v)] and [(.sub.J6)CH.sub.3OHH.sub.2OHOAc, 95+3+2 (v/v/v)]. The starting mobile phase condition was 7% B, 3 min; subsequently, ramp solvent B to 37.6% for 57 min and to 100% B, 3 min thereafter. The FLD was operated at X.sub.excitation=230 nm X.sub.emission=321 nm. Results are expressed in mg/g. In FIG. 2 is presented the chromatogram of the reference material NIST2381, DP1-DP10. NIST2381, DP1-DP10. FIG. 2 shows the chromatograpic profile by HPLC for the reference material DETERMINATION OF FAVANOLS ((+)-CATECHIN AND ()-EPICATECHIN) AND XANTHINES (THEOBROMINE AND CAFFEINE) CONTENT OF CACAO FLAVANOL EXTRACTS BY NORMAL PHASE HIGH-PERFORMANCE LIQUID

Chromatography-FLD/DAD Based Method

Chemicals

[0110] Theobromine, caffeine, (+)-Catechin and ()-Epicatechin were obtained from Sigma-Aldrich Co. (St. Louis, USA). Analytical grade reagents, such as solvents, were all chromatographic grade provided by Merck Millipore Co. (Darmstadt, Germany).

Sample Preparation

[0111] To 1 g of dry extract was added 15 mL of extracted solution (Isopropanol/water 60:40; water pH: 9). Then the mixture was submitted to sonic bath to enhancing the extraction during 1 h at room temperature. The resulting solution was vortexing for 1 min and incubating for 1 h at 20 C. The obtained product was centrifuged (4000 rpm; 20 min) and 1 mL of supernatant was filtered (0.45 gm). An aliquot of 200 gL was added with a solution of acetic glacial 0.1% in a volumetric ball (2 mL) and 5 gL was injected in the HPLC.

High Performance Liquid Chromatography-FLD/DAD

[0112] The analytical method applied allows detecting and quantifying the flavanoles and xanthines content in fermented and/or roasted cacao beans. The identification, integration and quantification of the chromatographic signals were performed with an HPLC coupling with two detectors online, DAD (diode array detector) and FLD (fluorescence detector) (Agilent 1200). The FLD was operated at X.sub.excit.sub.ation=280 nm X.sub.emission=315 nm and the DAD was operated at 280 nm. The quantification was performed by using the external standard method. The calibration curves were built for each standard as follows: 0.5-20 ppm for catechin; 5-200 ppm for epicatechin; 10-275 ppm for theobromine and 5-125 ppm for caffeine. The column used was a C18 Zorbax bonus HPLC Column (5 p,m particle size, L*I.D. 25 cm*4.6 mm). The results are expressed in mg/g.

TABLE-US-00006 TABLE 6 Compounds evaluated Compound Formula Chemical nomenclature Theobromine C.sub.7H.sub.8N.sub.4O.sub.2 3,7-dimetilxanthine Caffeine C.sub.8H.sub.10N.sub.4O.sub.2 1,3,7-trimetilxanthine (+)-Catechin C.sub.15H.sub.14O.sub.6 (2 R,3 S)-Catechin ()-Epicatechin C.sub.15H.sub.14O.sub.6 ()-Epicatechin (2 R,3 R)

Results are Summarized in Table 7.

[0113]

TABLE-US-00007 TABLE 7 Xanthines, flavanols and procyanidins in cacao extract (mg/g) Cacao extract Concentration (mg/g) Compound M SD Theobromine 14.6 0.2 Caffeine 2.30 0.03 (+)-Catechin 6.52 0.15 ()-Epicatechin 21.30 0.43 Total Procyanidins (DP1-DP10) 35.3
Additional extracts that can be used in the present invention includes:
Vitis vinifera Seeds and Skin Extract

[0114] Bioactive compounds: Vitis vinifera seeds standardized extracts contain approximately 15% of (+)-catechin and ()-epicatechin, and 80% of proanthocyanidins of ()-epicatechin 3-O-gallate, dimers, trimers, tetramers and their gallates and 5% of pentamers, hexamers, heptamers and their gallates.

[0115] Biological activity: Extracts have been tested in humans showing ability to reduce dyslipidemia markers, reduce blood pressure, reduce oxidative stress on LDL, help with weight management, improve skin conditions like chloasma, improve metabolic syndrome.

Persea americana Leaves, Peel and Seed Extract

[0116] Bioactive compounds: Flavanol monomers (catechin), proanthocyanidins, hydroxy-cinnamic acids (5-O-caffeoylquinic, 3-O-caffeoylquinic acid, 3-O-p-coumaroylquinic acid), and flavonol glycosides (quercetin derivatives) and procyanidin A trimers.

[0117] Biological activity: Avocado seeds may improve hypercholesterolemia, and be useful in the treatment of hypertension, hepatic inflammatory conditions and diabetes. Other activities are reported like amoebicidal, giardicidal, antimycobacterial, and antimicrobial.

Allium cepa Extract

[0118] Bioactive compounds: quercetin and quercetin glucosides, isorhamnetin glucosides, kaempferol glucoside, and, among anthocyanins, cyanidin glucoside. Also organosulphur compounds alliin. Biological activity: onion peel extract has the potential target in obesity by remodeling the characteristics of white fat to brown fat and controlling body weight. Promotes wound healing and improves the cosmetic appearance of postsurgical and hypertrophic scars. Also useful in the treatment of mild hypertension.

Allium sativum Extract

[0119] Bioactive compounds: quercetin and quercetin glucosides, isorhamnetin glucosides, kaempferol glucoside, and, among anthocyanins, cyanidin glucoside. Also organosulphur compounds allicin, ajoene, allicin, thiosulfinate, diallyl-di sulfide.

[0120] Biological activity: supported by clinical data as an adjuvant to dietetic management in the treatment of hyperlipidemia, and in the prevention of atherosclerosis. Also useful in the treatment of mild hypertension. Reduce symptoms associated with diabetes mellitus. Prevent inflammatory processes associated with asthma.

Vaccinium oxycoccos and Vaccinium macrocarpon Extract

[0121] Bioactive compounds: Proanthocyanidins (delphinidin), epicatechin, myricetin, and quercetin, chlorogenic and p-coumaric acid.

[0122] Biological activity: Cranberry fruit extracts (peel, seeds, pulp) may reduce the risk of symptomatic urinary tract infections in men and women.

Nasturtium Officinale Extract

[0123] Bioactive compounds: Phenyl isothiocyanates (PEITC), rutin.

[0124] Biological activity: Watercress plant extracts has been shown to reduce serum glucose, total cholesterol and LDL-cholesterol in diabetic rats, is also anti-inflammatory and antioxidant.

Petroselinum crispum Extract

[0125] Bioactive compounds: Apigenin, apigenin-7-O-glucoside or cosmosiin, apigenin-7-O-apiosyl-(1-->2)-O-glucoside or apiin and the coumarin 2,3-dihydroxyfuranocoumarin or oxypeucedanin hydrate.

[0126] Biological activity: Parsley extract has been shown to be a good antioxidant activity, reduce hepatic steatosis in animal models.

Vitis vinifera Fruit and Red Wine Extract

[0127] Bioactive compounds: 4.32 mg epicatechin, 2.72 mg catechin, 2.07 mg gallic acid, 0.9 mg trans-resveratrol, 0.47 mg rutin, 0.42 mg epsilon-viniferin, 0.28 mg, p-coumaric acid, 0.14 mg ferulic acid and 0.04 mg quercetin per gram.

[0128] Biological activity: For its antioxidant activity improve endothelial function in patients with coronary heart disease. Also improves markers of cardiovascular disease.

Citrus Peel Extracts:

[0129] Citrus reticulate, Citrus sinensis, Citrus limon and Citrus paradise.

[0130] Bioactive compounds: Hesperidin, neohesperidin, narirutin, tangeretin, sinnesetina, nobiletin. Caffeic acid, p-coumaric acid, ferulic acid and sinapic acid.

[0131] Biological activity: A wide range of biological effects have been published for molecules derived from citrus peel extracts. For hesperetin (flavanone) studies showed the effect of dietary hesperetin on the hepatic lipid content and enzyme activities involved in triacylglycerol synthesis in rats. Hesperetin and naringenin improves coronary vasodilatation, decrease the platelets activity to clot the blood, and prevents LDLs from oxidizing. Hesperidin has anti-inflammatory activity in vitro. In neuroptrotection, hesperetin protects cortical neurons from oxidative injury. Hesperidin and neohesperidin at physiological (0.4-4.0 {circumflex over ()}M) and high (20-50 {circumflex over ()}M) doses, all exhibit multiple mechanisms of neuroprotection against oxidative damage in PC12 cells, including the inhibition of ROS formation and caspase-3 activity, decreases in membrane and DNA damage, enhancement of antioxidant enzyme activity, and the maintenance of calcium homeostasis and mitochondrial potential. Naringenin has shown antimutagenic effect. Nobiletin and neohesperetin inhibit amylase-catalyzed starch digestion, while nobiletin inhibits both amylose and amylopectin digestion, which suggest an hypoglycemic effect.

Olea europaea Leaves and Fruits Extract

[0132] Bioactive compounds: Gallic acid, hydroxytyrosol, chorogenic acid, protocatechuic acid hydroxyphenylacetic acid, 4-Hydroxybenzoic acid, catechin, oleuropeine, p-coumaric acid, ferulic acid, rosmarinic acid, vanillic acid, m-coumaric acid, phenylacetic acid, cinnamic acid, luteolin, apigenin and 3-Hydroxybenzoic acid.

[0133] Biological activity: The olive oil polyphenols (standardised by the content of hydroxytyrosol and its derivatives) protect LDL particles from oxidative damage which contributing with cardiovascular health. The active components have a potent antioxidant activity.

Garnicia mangostana Extract

[0134] Bioactive compounds: a mangostin 0.20% (w/w), x-mangostin 0.11% (w/w). P-mangostin, 9-hydroxycalabaxanthone, mangostanol, mangostenone, allanxanthone E, mangostingone, garcinone D, mangosenone G, cudraxanthon, 1,5,8-trihydroxy-3-methoxy-2-(3-methylbut-2-enyl) xanthone, 8-deoxygartanin, gartanin, and smeathxanthone A.

[0135] Biological activity: Garcinia mangostana improves the antioxidant activity of plasma in humans and has antiinflamatory activity.

Garcinia Species Extract: G. cambogia, G. kola, G. madruno

[0136] Bioactive compounds: biflavonoids, flavonoids, benzophenones, xanthones, and organic acids, hydroxycitric acid.

[0137] Biological activity: Garcinia species are used for the prevention and treatment of multiple symptoms and diseases such as ulcers, diarrhoea, hypertension, obesity, inflammatory disorders, hepatic damage, among others.

Thermal Characterization of Cacao Extract

[0138] Cacao extracts were analyzed by a variety of thermal characterization techniques.
Cacao Extract Differential Scanning calorimetry (DSC) characterization: First heat thermal analysis is shown in FIG. 3.

[0139] The melting points of different fatty acids are shown in Table 8.

TABLE-US-00008 TABLE 8 Unsaturated Fatty Acids Melting Formula Common Name Point CH3(CH2)5CHCH(CH2)7CO2H Palmitoleic Acid 0 C. CH3(CH2)7CHCH(CH2)7CO2H Oleic acid 13 C. CH3(CH2)4CHCHCH2CHCH(CH2)7CO2H Linoleic Acid 5 C. CH3CH2CHCHCH2CHCHCH2CHCH(CH2)7CO2H Linoleic acid 11 C. CH3(CH2)4(CHCHCH2)4(CH2)2CO2H Arachidonic acid 49 C.

[0140] FIG. 4 shows Cacao extract Differential Scanning calorimetry (DSC) characterization: Cooling curve, while FIG. 5 features a cacao extract Differential Scanning calorimetry (DSC) characterization second heat thermal analysis.

[0141] FIG. 6 describes the thermogravimetric analysis (TGA) characterization of a cacao extract while FIG. 7 is a cacao extract Isothermal Thermogravimetric characterization.

[0142] FIG. 8 illustrates the cacao extract Oxygen Induction Time (OIT) characterizationStability Analysis and FIG. 9 shows a cacao extract Fourier-Transform Infrared Spectrocopy (FTIR) characterization.

[0143] The FTIR spectrum was compared to other spectra of cacao extracts in the literature, showing similar absorption bands. [Vesela A, eta al. Infrared spectroscopy and outer product analysis for quantification of fat, nitrogen, and moisture of cacao powder. Analytical Chimica Acta 2007; 601: 77-86]

[0144] The absorption bands in the cacao extract (FTIR) are shown in Table 9.

TABLE-US-00009 TABLE 9 Wave number (cm.sup.1) Assignment 3362 Stretching OH of fatty acids, carbohydrates and others 2921 Stretching CH of methylene and CH2 carbohydrates 2852 Stretching CH of fatty Acids 1740 Stretching CO of fatty Acids 1656 Stretching CO of amide I protein 1525 Stretching CN of amide II protein 1448 Flexing CH 138 Flexing OH of fatty acids, carbo- hydrates and others 1248 Flexing NH of amide III protein 1152 Stretching CO of fatty acids 1024 Stretching CO of carbohydrates <900 Flexing CH out of scope

Thermal Characterization of ()-Epicatechin

[0145] FIG. 10 illustrates the ()-Epicatechin at 90% Differential Scanning calorimetry (DSC) characterization for the first heating thermal analysis.

Thermal Characterization of Cocoa Extract

[0146] FIG. 11 illustrates the cocoa xxtract Differential Scanning calorimetry (DSC) characterization first heating thermal analysis.

EXAMPLES

[0147] The invention is exemplified as shown in the Examples below.

Example I

50EXT-40SOLU-10L100-T120-120 RPM-RT

50% Cacao Extract

[0148] 40% Soluplus (BASF polymer)
10% Eudragit L100 (Evonik polymer)
Process Conditions: batch melt mixing at 120 RPM and processing temperature of 120 C.

Example II

50EXT-30AN7-10SR-10L100-T150-120 RPM-RT

50% Cacao Extract

[0149] 30% Aqualon N7 (Ashland polymer)
10% Kollidon SR (Basf polymer)
10% Eudragit L100 (Evonik polymer)
Process Conditions: batch melt mixing at 120 RPM and processing temperature of 150 C.

Example III

50EXT-30AN7-10SR-10L100-120 RPM-TSE

50% Cacao Extract

[0150] 30% Aqualon N7 (Ashland polymer)
10% Kollidon SR (Basf polymer)
10% Eudragit L100 (Evonik polymer)
Process Conditions: twin screw extrusion (Nano 16 Leistritz) at 120 RPM (co-rotating screws) and temperature profile: 80 C. (feed zone), 150 C., 140 C. (metering zone), 140 C. (die)

Example IV

50EXT-30ES10-10SR-10L100-T150-120 RPM-RT

50% Cacao Extract

[0151] 30% Ethocel Standard 10 (Dow polymer)
10% Kollidon SR (Basf polymer)
10% Eudragit L100 (Evonik polymer)
Process Conditions: batch melt mixing at 120 RPM and 150 C.

Example V

50EXT-30ES10-10SR-10L100-120 RPM-TSE

50% Cacao Extract

[0152] 30% Ethocel Standard 10 (Dow polymer)
10% Kollidon SR (Basf polymer)
10% Eudragit L100 (Evonik polymer)
Process Conditions: twin screw extrusion (Nano 16 Leistritz) at 120 RPM (co-rotating screws) and temperature profile: 80 C. (feed zone), 140 C., 140 C. (metering zone), 130 C. (die). FIG. 12 shows the Release profile of polyphenols in artificial saliva at 23 C. for EXAMPLES I TO V.

[0153] The FIG. 12 graph shows the encapsulated cacao extract (batch melt mixing, RT, and Twin Screw Extrusion, TSE), not tasted in food.

The Artificial Saliva at pH 6.2 is shown in Table 10.

TABLE-US-00010 TABLE 10 Chemical Amount (g/L) CaCl22H2O 0.228 MgCl26H2O 0.061 NaCl 1.071 K2CO3 0.603 Na2HPO4 0.204 NaH2PO4 0.273

[0154] FIG. 13 shows the release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. Releases for EXAMPLES I TO V.

[0155] FIG. 14 illustrates the Oxygen Induction Time (OIT) for stability analysis of encapsulated formulations vs cacao extract for EXAMPLES I TO V.

Example VI

50EXT-30AN7-10 SR-10L100-120 RPM-TSE

50% Cacao Extract

[0156] 30% Aqualon N7 (Ashland polymer)
10% Kollidon SR (Basf polymer)
10% Eudragit L100 (Evonik polymer)
Process Conditions: twin screw extrusion (Nano 16 Leistritz) at 120 RPM (co-rotating screws) and temperature profile: 80 C. (feed zone), 150 C., 140 C. (metering zone), 140 C. (die).

[0157] FIG. 15 shows the release profile of polyphenols in artificial saliva at 23 C. for EXAMPLE VI with particle size distribution between 250 m and 425 m (Mesh 40), 180 m and 250 m (Mesh 60), 125 m and 180 m (Mesh 80), and particle size smaller than 120 m (Mesh 120):

Example VII

50EXT-30ES10-10SR-10L100-120 RPM-TSE

50% Cacao Extract

[0158] 30% Ethocel Standard 10 (Dow polymer)
10% Kollidon SR (Basf polymer)
10% Eudragit L100 (Evonik polymer)
Process Conditions: twin screw extrusion (Nano 16 Leistritz) at 120 RPM (co-rotating screws) and temperature profile: 80 C. (feed zone), 140 C., 140 C. (metering zone), 130 C. (die).

[0159] FIG. 16 describes the Thermogravimetric analysis of cacao extract vs an encapsulated formulation from example VII. Encapsulation provides thermal stability and moisture resistance to the cacao extract.

Example VIII

50EXT-50ES10-T180-120 RPM-RT

50% Cacao Extract

[0160] 50% Ethocel Standard 10 (Dow polymer)
Process Conditions: batch melt mixing at 120 RPM and processing temperature of 180 C.

Example IX

50EXT-50AN7-T140-70 RPM-RT

50% Cacao Extract

[0161] 50% Aqualon N7 (Ashland polymer)
Process Conditions: batch melt mixing at 70 RPM and processing temperature of 140 C.

[0162] FIG. 17 illustrates Release profile of polyphenols in artificial saliva at 23 C. for EXAMPLE VIII and IX.

[0163] FIG. 18 shows the release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. for examples VIII and IX.

Example X

50EPI-30AN7-10L100-10 SR-T130-70 RPM-RT

50% ()-Epicatechin (90%)

[0164] 30% Aqualon N7 (Ashland polymer)
10% Eudragit L100 (Evonik polymer)
10% Kollidon SR (Basf polymer)
Process Conditions: batch melt mixing at 70 RPM and processing temperature of 130 C.

Example XI

50EPI-30AN7-10L100-10SR-T130-100RPM-RT

50% ()-Epicatechin (90%)

[0165] 30% Aqualon N7 (Ashland polymer)
10% Eudragit L100 (Evonik polymer)
10% Kollidon SR (Basf polymer)
Process Conditions: batch melt mixing at 100 RPM and processing temperature of 130 C.

Example XII

50EPI-30AN7-10L100-10 SR-T150-70 RPM-RT

50% ()-Epicatechin (90%)

[0166] 30% Aqualon N7 (Ashland polymer)
10% Eudragit L100 (Evonik polymer)
10% Kollidon SR (Basf polymer)
Process Conditions: batch melt mixing at 70 RPM and processing temperature 150 C.

Example XIII

50EPI-30AN7-10L100-10SR-T150-100RPM-RT

50% ()-Epicatechin (90%)

[0167] 30% Aqualon N7 (Ashland polymer)
10% Eudragit L100 (Evonik polymer)
10% Kollidon SR (Basf polymer)
Process Conditions: batch melt mixing at 100 RPM and processing temperature 150 C.

[0168] FIG. 19 describes the torque and melt temperature behavior of examples X to XIII

[0169] FIG. 20 describes the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for examples X to XIII with a particle size distribution between 250 m and 425 m (Mesh 40).

[0170] FIG. 21 illustrates the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for EXAMPLE X to XIII with a particle size distribution between 180 m and 250 m (Mesh 60).

[0171] FIG. 22 features release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. Releases for examples X to XIII with a particle size distribution between 250 m and 425 m (Mesh 40); and particle size distribution between 180 m and 250 m (Mesh 60).

[0172] Table 11 below describes the release profile percentages for EXAMPLES X to XIII with a particle size distribution between 250 m and 425 m (Mesh 40); and particle size distribution between 180 m and 250 m (Mesh 60).

TABLE-US-00011 TABLE 11 Time 30 s 60 s 120 s 1 h 2 h 3 h 4 h % % % % % % % Samples release release release release release release release EPICATECHIN AT 75.30 88.70 93.43 98.59 99.94 100.22 100.84 90% 50EPI-30AN7-10SR- 7.01 10.36 14.35 61.80 71.78 88.41 90.23 10L100- T130- 70 rpm- RT- Mesh 40 50EPI-30AN7-10SR- 4.77 6.33 11.72 50.23 60.80 77.08 80.09 10L100- T130- 100 rpm- RT-Mesh 40 50EPI-30AN7-10SR- 4.97 10.39 14.15 48.25 58.27 83.80 87.44 10L100- T150- 70 rpm- RT-Mesh 40 50EPI-30AN7-10SR- 7.09 10.36 12.86 43.92 52.10 78.20 80.73 10L100- T150- 100 rpm- RT-Mesh 40 50EPI-30AN7-10SR- 9.50 16.09 26.67 75.01 78.11 88.15 89.23 10L100- T130- 70 rpm- RT- Mesh 60 50EPI-30AN7-10SR- 8.35 14.50 17.79 66.27 73.17 79.74 85.23 10L100- T130- 100 rpm- RT-Mesh 60 50EPI-30AN7-10SR- 8.10 10.36 16.71 73.63 74.51 79.77 85.33 10L100- T150- 70 rpm- RT-Mesh 60 50EPI-30AN7-10SR- 8.93 11.87 20.26 71.05 77.03 82.41 87.15 10L100- T150- 100 rpm- RT-Mesh 60

Example XIV

50EPI-30ES10-10L100-10SR-T130-70 RPM-RT

50% ()-Epicatechin (90%)

[0173] 30% Ethocel Standard 10 (Dow polymer)
10% Eudragit L100 (Evonik polymer)
10% Kollidon SR (Basf polymer)
Process Conditions: batch melt mixing at 70 RPM and processing temperature of 130 C.

Example XV

50EPI-30ES10-10L100-10SR-T130-100 RPM-RT

50% ()-Epicatechin (90%)

[0174] 30% Ethocel Standard 10 (Dow polymer)
10% Eudragit L100 (Evonik polymer)
10% Kollidon SR (Basf polymer)
Process Conditions: batch melt mixing at 100 RPM and processing temperature of 130 C.

Example XVI

50EPI-30ES10-10L100-10SR-T150-70 RPM-RT

50% ()-Epicatechin (90%)

[0175] 30% Ethocel Standard 10 (Dow polymer)
10% Eudragit L100 (Evonik polymer)
10% Kollidon SR (Basf polymer)
Process Conditions: batch melt mixing at 70 RPM and processing temperature 150 C.

Example XVII

50EPI-30ES10-10L100-10SR-T150-100RPM-RT

50% ()-Epicatechin (90%)

[0176] 30% Ethocel Standard 10 (Dow polymer)
10% Eudragit L100 (Evonik polymer)
10% Kollidon SR (Basf polymer)
Process Conditions: batch melt mixing at 100 RPM and processing temperature 150 C.

[0177] FIG. 23 illustrates the torque and melt temperature behavior of examples XIV to XVII.

[0178] FIG. 24 shows the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for examples X to XIII with a particle size distribution between 250 m and 425 m (Mesh 40).

[0179] FIG. 25 features the release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. Releases for EXAMPLES X to XIII with a particle size distribution between 250 m and 425 m (Mesh 40).

[0180] Table 12 below shows the dissolution release percentages for EXAMPLES X to XIII with a particle size distribution between 250 m and 425 m (Mesh 40).

TABLE-US-00012 TABLE 12 Time 30 s 60 s 120 s 1 h 2 h 3 h 4 h % % % % % % % Samples release release release release release release release 50EPI-30ES10-10SR-10L100- 6.31 9.17 12.78 31.35 39.79 69.97 70.99 T130- 70 rpm-RT 50EPI-30ES10-10SR-10L100- 7.52 10.69 14.35 33.38 42.74 76.81 82.44 T130- 100 rpm-RT 50EPI-30ES10-10SR-10L100- 6.97 10.27 14.14 41.80 49.60 69.95 70.09 T150- 70 rpm-RT 50EPI-30ES10-10SR-10L100- 6.70 9.52 12.88 31.12 39.36 73.36 79.32 T150- 100 rpm-RT

Example XVIII

50EPI-30AN7-10L100-10SR-T160-250RPM-AS70-C2.3

50% ()-Epicatechin (90%)

[0181] 30% Aqualon N7 (Ashland polymer)
10% Eudragit L100 (Evonik polymer)
10% Kollidon SR (Basf polymer)
Process Conditions: twin screw extrusion (Nano 16 Leistritz) at 250 RPM (co-rotating screws), temperature profile: 160 C. (feed zone), 165 C., 145 C. (metering zone), 145 C. (die); feeding rate of volumetric feeder at 680 g/h; and estimated filling factor 33%.

[0182] FIG. 26 illustrates the torque and melt temperature behavior of example XVIII.

[0183] FIG. 27 shows the thermal characterization of EXAMPLE XVIII.

[0184] FIG. 28 describes the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for example XVIII with particle size distribution between 250 m and 425 m (Mesh 40), particle size distribution between 180 m and 250 m (Mesh 60), particle size distribution between 125 m and 180 m (Mesh 80), and particle size smaller than 125 m (Mesh 120).

[0185] FIG. 29 features the release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. Releases for EXAMPLE XVIII with particle size distribution between 250 m and 425 m (Mesh 40), particle size distribution between 180 m and 250 m (Mesh 60), particle size distribution between 125 m and 180 m (Mesh 80), and particle size smaller than 125 m (Mesh 120).

[0186] Table 13 illustrates the release profile percentages for EXAMPLE XVIII with particle size distribution between 250 m and 425 m (Mesh 40), particle size distribution between 180 m and 250 m (Mesh 60), particle size distribution between 125 m and 180 m (Mesh 80), and particle size smaller than 125 m (Mesh 120).

TABLE-US-00013 TABLE 13 Time 30 s 60 s 120 s 1 h 2 h 3 h 4 h Samples % % % % % % % ()-EPICATECHIN AT 90% 75.30 88.70 93.43 98.59 99.94 100.22 100.84 50EPI-30AN7-10L100-10SR-T160- 4.28 4.99 5.53 28.71 44.11 88.00 88.02 250 RPM-AS70-C2.3-Mesh 40 50EPI-30AN7-10L100-10SR-T160- 5.93 7.59 9.75 49.01 66.04 89.02 90.58 250 RPM-AS70-C2.3-Mesh 60 50EPI-30AN7-10L100-10SR-T160- 6.02 7.86 11.37 67.72 89.55 90.75 93.00 250 RPM-AS70-C2.3-Mesh 80 50EPI-30AN7-10L100-10SR-T160- 12.10 16.28 18.23 84.95 87.62 93.71 95.93 250 RPM-AS70-C2.3-Mesh 120

Example XIX

50EPI-30ES10-10L100-10SR-T160-270 RPM-AS70-C2.3

Epicatechin (90%)

[0187] 30% Ethocel Standard 10 (Dow polymer)
10% Eudragit L100 (Evonik polymer)
10% Kollidon SR (Basf polymer)
Process Conditions: twin screw extrusion (Nano 16 Leistritz) at 250 RPM (co-rotating screws), temperature profile: 160 C. (feed zone), 165 C., 145 C. (metering zone), 145 C. (die); feeding rate of volumetric feeder at 680 g/h; and estimated filling factor 31%.

[0188] FIG. 30 describes the torque and melt temperature behavior of example XIX.

[0189] FIG. 31 illustrates the thermal characterization of example XIX.

[0190] FIG. 32 shows the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for example XIX with particle size distribution between 250 m and 425 m (Mesh 40), particle size distribution between 180 m and 250 m (Mesh 60), particle size distribution between 125 m and 180 m (Mesh 80), and particle size smaller than 125 m (Mesh 120).

[0191] FIG. 33 describes the release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. Releases for example XIX with particle size distribution between 250 m and 425 m (Mesh 40), particle size distribution between 180 m and 250 m (Mesh 60), particle size distribution between 125 m and 180 m (Mesh 80), and particle size smaller than 125 m (Mesh 120).

[0192] Table 14 shows the release profile percentages for EXAMPLE XIX with particle size distribution between 250 m and 425 m (Mesh 40), particle size distribution between 180 m and 250 m (Mesh 60), particle size distribution between 125 m and 180 m (Mesh 80), and particle size smaller than 125 m (Mesh 120):

TABLE-US-00014 TABLE 14 Time 30 s 60 s 120 s 1 h 2 h 3 h 4 h Samples % % % % % % % ()-EPICATECHIN AT 90% 75.30 88.70 93.43 98.59 99.94 100.22 100.84 50EPI-30ES10-10L100-10SR-T160- 4.78 5.89 6.81 29.53 40.44 71.69 71.83 270 RPM-AS70-C2.3-Mesh 40 50EPI-30ES10-10L100-10SR-T160- 5.37 7.37 8.84 35.82 48.34 77.21 82.15 270 RPM-AS70-C2.3-Mesh 60 50EPI-30ES10-10L100-10SR-T160- 7.21 9.72 13.31 69.01 73.67 78.77 83.53 270 RPM-AS70-C2.3-Mesh 80 50EPI-30ES10-10L100-10SR-T160- 7.81 12.38 14.42 71.19 76.57 84.52 85.77 270 RPM-AS70-C2.3-Mesh 120

Example XX

50EXT-30ES10-10L100-10SR-T155-400RPM-AS70-C2.3

50% Cocoa Extract

[0193] 30% Ethocel Standard 10 (Dow polymer)
10% Eudragit L100 (Evonik polymer)
10% Kollidon SR (Basf polymer)
Process Conditions: twin screw extrusion (Nano 16 Leistritz) at 400 RPM (co-rotating screws) and temperature profile: 165 C. (feed zone), 150 C., 140 C. (metering zone), 140 C. (die); feeding rate of volumetric feeder at 990 g/h, and estimated filling factor 26%.

[0194] FIG. 34 illustrates the torque and melt temperature behavior of example XX.

[0195] FIG. 35 features the thermal characterization of example XX.

[0196] FIG. 36 illustrates the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for example XX with particle size distribution between 250 m and 425 m (Mesh 40), particle size distribution between 180 m and 250 m (Mesh 60), particle size distribution between 125 m and 180 m (Mesh 80), and particle size smaller than 125 m (Mesh 120).

[0197] FIG. 37 describes the release profile of epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. Releases for example XX with particle size distribution between 250 m and 425 m (Mesh 40), particle size distribution between 180 m and 250 m (Mesh 60), particle size distribution between 125 m and 180 m (Mesh 80), and particle size smaller than 125 m (Mesh 120).

[0198] Table 15 features the release profile percentages for EXAMPLE XX with particle size distribution between 250 m and 425 m (Mesh 40), particle size distribution between 180 m and 250 m (Mesh 60), particle size distribution between 125 m and 180 m (Mesh 80), and particle size smaller than 125 m (Mesh 120)

TABLE-US-00015 TABLE 15 Time 30 s 60 s 120 s 1 h 2 h 3 h 4 h Samples % % % % % % % Cocoa Extract 33.53 51.86 78.86 103.48 105.51 105.90 106.91 50EXT-30ES10-10L100-10SR- 7.12 12.83 12.83 36.48 47.65 55.36 57.59 T155-400 RPM-AS70-C2.3- Mesh 40 50EXT-30ES10-10L100-10SR- 7.59 10.07 17.37 44.61 58.82 62.44 77.60 T155-400 RPM-AS70-C2.3- Mesh 60 50EXT-30ES10-10L100-10SR- 15.00 16.38 22.10 48.67 63.89 64.87 75.58 T155-400 RPM-AS70-C2.3- Mesh 80 50EXT-30ES10-10L100-10SR- 11.75 14.21 26.83 55.77 62.88 64.46 65.07 T155-400 RPM-AS70-C2.3- Mesh 120

Example XXI

50EXT-30AN7-10L100-10 SR-T155-400 RPM-AS73-C2.3

50% Cocoa Extract

[0199] 30% Aqualon N7 (Ashland polymer)
10% Eudragit L100 (Evonik polymer)
10% Kollidon SR (Basf polymer)
Process Conditions: twin screw extrusion (Nano 16 Leistritz) at 400 RPM (co-rotating screws) and temperature profile: 165 C. (feed zone), 145 C., 140 C. (metering zone), 140 C. (die), feeding rate of volumetric feeder at 1.04 kg/h, and estimated filling factor 27%.

[0200] FIG. 38 shows the torque and melt temperature behavior of example XXI.

[0201] FIG. 39 describes the thermal characterization of example XXI.

[0202] FIG. 40 illustrates the release profile of ()-Epicatechin in artificial saliva at pH 6.2 and 23 C. for example XXI with particle size distribution between 250 m and 425 m (Mesh 40), particle size distribution between 180 m and 250 m (Mesh 60), particle size distribution between 125 m and 180 m (Mesh 80), and particle size smaller than 125 m (Mesh 120).

[0203] FIG. 41 shows the release profile of ()-Epicatechin at 2 hour in a medium with pH of 1.2 and 2 more hours in a medium with pH of 6.8, at 37 C. Releases for example XXI with particle size distribution between 250 m and 425 m (Mesh 40), particle size distribution between 180 m and 250 m (Mesh 60), particle size distribution between 125 m and 180 m (Mesh 80), and particle size smaller than 125 m (Mesh 120).

[0204] Table 16 illustrates the dissolution release percentages for EXAMPLE XXI with particle size of 425 m (Mesh 40), 250 m (Mesh 60), 180 m (Mesh 80), and 125 m (Mesh 120):

TABLE-US-00016 TABLE 16 Time 30 s 60 s 120 s 1 h 2 h 3 h 4 h Samples % % % % % % % Cocoa Extract 33.53 51.86 78.86 103.48 105.51 105.90 106.91 50EXT-30AN7-10L100- 4.95 7.12 9.88 37.50 46.64 68.50 71.54 10SR-T155-400 RPM- AS73-C2.3-Mesh 40 50EXT-30AN7-10L100- 5.38 8.30 15.00 42.58 45.62 72.55 75.58 10SR-T155-400 RPM- AS73-C2.3-Mesh 60 50EXT-30AN7-10L100- 5.54 9.88 16.97 48.67 61.86 61.43 64.46 10SR-T155-400 RPM- AS73-C2.3Mesh 80 50EXT-30AN7-10L100- 13.50 23.08 27.81 61.86 66.94 66.48 69.52 10SR-T155-400 RPM- AS73-C2.3Mesh 120

Example XXII

[0205] Chocolate Candy Production Using Nutraceutical Compound 50EPI-30AN7-10L100-10SR-T160-250 rpm-AS70-C2.3in Mesh 120 for 200 mg of Polyphenols/Unit of Chocolate Candy

TABLE-US-00017 TABLE 16 Lower limit Upper limit Ingredients [w/w %] [w/w %] Sucrose 20% 30% Liquor, solids 50% 55% and Colombian origin cocoa butter Essence 0.1% 1.0% Nutraceutical com- 0.2% 0.4% pound 10193-120
Nutraceutical compound 10193-120 in equivalent to: 50EPI-30AN7-10L100-10SR-T160-250 rpm-AS70-C2.3in MESH 120(Example XVIII)

Manufacturing Process of Chocolate Candy:

[0206] 1. Blending of ingredients (Nutraceutical compound included)
2. Refining process: <30 microns. 22-35 C.
3. Conching process per 24 hours, Rolls temperature: 60 C.
4. Tempering chocolate process
5. Molding chocolate
6. Cooling chocolate
7. Demolding chocolate
8. Packaging chocolate
Result of Organoleptic Panel: Test 1, Good texture, melts well in the mouth, presents creaminess or pleasant fat sensation, good micrage. The total impression is 3, although it is different from the pattern in descriptors. Not significant differences in bitter taste was perceived.

TABLE-US-00018 TABLE 17 Name of product: Chocolate cover 70% Test Taste profile Sample preparation Pilot plant. Differences are expected Sample 1 10193-120 Descriptor Example XXII Pattern Sweet taste 2.5 3 Chocolate taste 4.5 4.5 Bitter taste 3 2.5 Vanilla taste 0 1 Nut taste 1 0 Green taste 0 1.5 Dry grass taste 1.5 0 Fruity taste 0 1 Floral taste 0 1.5 Astringent taste 2 3 Total impression 3 3 *Total scale impression or general quality: 1: Low; 2: Medium; 3: High
FIG. 42 is a comparative radar graphic for taste profile in chocolate candy evaluated (examples XXII)

Example XXIII

[0207] Cereal Bar #1 Production Using Nutraceutical Compound 50EPI-30AN7-10L100-10SR-T160-250 rpm-AS70-C2.3in Mesh 120 for 90 mg of ()-Epicatechin/Unit of Cereal Bar

TABLE-US-00019 Lower limit Upper limit Ingredients [w/w %] [w/w %] Binder 40% 60% Polysaccharides Polyols Milk matrix Solid 45% 55% Assorted cereals Nutraceutical compound 1% 2% 10193-120

Manufacturing Process:

[0208] Mixing the ingredients of the binder, and take it to a higher Brix than 50
Mix the binder, with the solid and Nutraceutical compound 10193-120
Formation of the bar. 80-95 C. (Process time: aprox 5 min)
Cooling below 14 C.

Cutting and Packaging

[0209] Nutraceutical compound 10193-120 in equivalent to: 50EPI-30AN7-10L100-10SR-T160-250 rpm-AS70-C2.3in MESH 120(Example XVIII)
Result of Organoleptic Panel: Excess binder, different texture, pale color of the binder. Not significant differences in bitter taste was perceived.

Example XXIV

[0210] Cereal Bar #2 Production Using Nutraceutical Compound 50EPI-30AN7-10L100-10SR-T160-250 rpm-AS70-C2.3in Mesh 40 for 90 mg of ()-Epicatechin/Unit of Cereal Bar

TABLE-US-00020 Lower limit Upper limit [w/w %] [w/w %] Binder 40% 60% Polysaccharides Polyols Milk matrix Solid 45% 55% Assorted cereals Nutraceutical compound 1% 2% 10193-40

Manufacturing Process:

[0211] Mixing the ingredients of the binder, and take it to a higher Brix than 50
Mix the binder, with the solid and Nutraceutical compound 10193-40
Formation of the bar. 80-95 C. (Process time: aprox 5 min)
Cooling below 14 C.

Cutting and Packaging

[0212] Nutraceutical compound 10193-40 in equivalent to: 50EPI-30AN7-10L100-10SR-T160-250 rpm-AS70-C2.3in MESH 40(Example XVIII)
Result of Organoleptic Panel: Less crispy, lack brightness, has a strange taste (PSH greaseMolding). Not significant differences in bitter taste was perceived.

Example XXV

[0213] Cereal Bar #3 Production Using Nutraceutical Compound 50EPI-30ES10-10L100-10SR-T160-270 rpm-AS70-C2.3in Mesh 120 for 90 mg of ()-Epicatechin/Unit of Cereal Bar

[0214] Ingredients

TABLE-US-00021 Lower limit Upper limit [w/w %] [w/w %] Binder 40% 60% Polysaccharides Polyols Milk matrix Solid 45% 55% Assorted cereals Nutraceutical compound 1% 2% 10195-120

Manufacturing Process:

[0215] Mixing the raw materials of the binder, and take it to a higher Brix than 50
Mix the binder, with the solid and Nutraceutical compound 10195-120
Formation of the bar. 80-95 C. (Process time: aprox 5 min)
Cooling below 14 C.

Cutting and Packaging

[0216] Nutraceutical compound 10195-120 in equivalent to: 50EPI-30ES10-10L100-10SR-T160-270 rpm-AS70-C2.3in MESH 120(Example XIX)
Result of Organoleptic Panel: Dry, it seems with less binder, more opaque. Not significant differences in bitter taste was perceived.

Example XXVI

[0217] Cereal Bar #4 Production Using Nutraceutical Compound 50EPI-30ES10-10L100-10SR-T160-270 rpm-AS70-C2.3in mesh 40 for 90 mg of ()-Epicatechin/Unit of Cereal Bar

TABLE-US-00022 Lower limit Upper limit Ingredients [w/w %] [w/w %] Binder 40% 60% Polysaccharides Polyols Milk matrix Solid 45% 55% Assorted cereals Nutraceutical compound 1% 2% 10195-40

Manufacturing Process:

[0218] Mixing the raw materials of the binder, and take it to a higher Brix than 50
Mix the binder, with the solid and Nutraceutical compound 10195-40
Formation of the bar. 80-95 C. (Process time: aprox 5 min)
Cooling below 14 C.
Cutting and packaging
Nutraceutical compound 10195-40 in equivalent to: 50EPI-30ES10-10L100-10SR-T160-270 rpm-AS70-C2.3in Mesh 40(Example XIX)
Result of Organoleptic Panel: Texture more similar to the pattern, appearance similar to the pattern, residual flavor, and fatty taste (apparently it is the fat used to portion). Not significant differences in bitter taste was perceived.

Example XXVII

[0219] Cereal Bar #5 Production Using Nutraceutical Compound 50EXT-30ES10-10L100-10SR-T155-400 rpm-AS70-C2.3in Mesh 40 for 90 mg of ()-Epicatechin/Unit of Cereal Bar

TABLE-US-00023 Lower limit Upper limit Ingredients [w/w %] [w/w %] Binder 40% 60% Polysaccharides Polyols Milk matrix Solid 45% 55% Assorted cereals Nutraceutical compound 20% 20% 10196-40

Manufacturing Process:

[0220] Mixing the raw materials of the binder, and take it to a higher Brix than 50
Mix the binder, with the solid and Nutraceutical compound 10196-40
Formation of the bar. 80-95 C. (Process time: aprox 5 min)
Cooling below 14 C.
Cutting and packaging
Nutraceutical compound 10196-40 in equivalent to: 50EXT-30ES10-10L100-10SR-T155-400 rpm-AS70-C2.3in MESH 40(Example XX)
Result of Organoleptic Panel: Sandy feeling, a lot of particle residue that is not pleasant. Not significant differences in bitter taste was perceived.

TABLE-US-00024 Product Name: Cereal Bar Test name Taste profile Samples preparation Manufacturing in Pilot Plant. Some differences are expected. Sample 1: Sample 2: Sample 3: Sample 4: Sample 5: 10193-40 10195-120 10195-40 10196-40 10193-120 Example Example Example Example Example Descriptor XXIV XXV XXVI XXVII XXIII Pattern Salty taste 0.5 0 0.5 0 0 1 Sweet taste 3.5 3 4 3.5 4.5 4 Dairy taste 1.5 1.5 2 0 1.5 2 Bitter taste 2 2 2 2.5 2.5 2 Cereal taste 3 3.5 3.5 3 2.5 3.5 Toasted taste 0 0 0 3 0 0 Caramel taste 0 0 0 0 0 0 Total 2 2 3 2 2 3 impression *Total scale impression or general quality: 1: Low; 2: Medium; 3: High

General Remarks

[0221] Sample 1, Less crispy, lack brightness, has a strange taste (PSH greaseMolding)
Sample 2, Dry, it seems with less binder, more opaque.
Sample 3, Texture more similar to the pattern, appearance similar to the pattern, residual flavor, and fatty taste (apparently it is the fat used to portion).
Sample 4, Sandy feeling, a lot of particle residue that is not pleasant.
Sample 5, Excess binder, different texture, pale color of the binder.
FIG. 43 is comparative radar graphic for taste profile in cereal bars evaluated (examples XXIII, XIV, XV, XVI, XVII)

Example XXVIII

Screw Configurations

[0222] ()-Epicatechin is degraded by light, temperature, residence time and excessive shear.
The possible degradation of the ()-Epicatechin during extrusion limits the operating conditions of the extruder, and possible screw configurations.
The best suitable screw should have a long feeding zone and short melting zone to control the melt temperature.
FIGS. 44 and 45 show two possible screw configurations.

[0223] Another option could be the use of an additional feeder for the cacao or cocoa extracts or pure ()-epicatechin. This set up requires two feeders: one for the polymers and one for the extracts. The feeders can be located as demanded by the cocoa or cacao extract. The polymer feeder should be placed close to the main hopper, and the second feeder (cocoa or cacao extract) should be placed after the melting zone.

[0224] The temperature profile should deliver a polymer melt below 160 C. as it was shown in the EXAMPLES XVIII, XIX, XX and XXI.

Screw configuration 2.3: D=16 mm, L/D=26.25

Screw Configuration 2.4: D=16 mm, L/D=32.81

[0225] With the above screw configurations and the reported operating conditions, the loss of ()-epicatechin during extrusion was reduced to values below 8.8% wt.: This assessment was measured by HPLC. The physical mixtures were used as reference values (See ()-Epicatechin content).

TABLE-US-00025 ()-Epicatechin EPI Loss in Samples Content [% w/w] process (% w/w) 50EPI-30AN7-10SR-10L100- 41.3 7.6 MESH 120 50EPI-30ES10-10SR-10L100- 40.6 8.8 MESH 120 50EPI-30AN7-10SR-10L100 44.7 Reference (PHYSICAL MIXTURE) value 50EPI-30ES10-10SR-10L100 44.5 Reference (PHYSICAL MIXTURE) value EPICATECHIN AT 90% 85.7 Reference value

[0226] All patents, patent applications and publications cited in this application including all cited references in those patents, applications and publications, are hereby incorporated by reference in their entirety for all purposes to the same extent as if each individual patent, patent application or publication were so individually denoted.

[0227] While the many embodiments of the invention have been disclosed above and include presently preferred embodiments, many other embodiments and variations are possible within the scope of the present disclosure and in the appended claims that follow. Accordingly, the details of the preferred embodiments and examples provided are not to be construed as limiting. It is to be understood that the terms used herein are merely descriptive rather than limiting and that various changes, numerous equivalents may be made without departing from the spirit or scope of the claimed invention.