WO2018147718A2

WO2018147718A2 - Cocoa pericarp compositions and method for producing same

Info

Publication number: WO2018147718A2
Application number: PCT/MX2018/000012
Authority: WO
Inventors: Guillermo Ceballos
Original assignee: Guillermo Ceballos
Priority date: 2017-02-13
Filing date: 2018-02-13
Publication date: 2018-08-16
Also published as: WO2018147718A3; CA3091105A1; WO2018147718A8

Abstract

The cocoa pod pericarp (CP) is used to produce a nutritional ingredient in the form of flour using convection heating, freeze-drying and or microwave drying. The dry CP physical and technological properties observed show that it can be potentially used as an ingredient of manufactured food products. The bioactive compound levels increased significantly in the dry CP samples, as compared with raw materials, especially in the case of freeze-drying methods. The cocoa pod waste material can be used in a commercially advantageous manner for the treatment of humans and animals in order to improve anthropometric measurements or cardiometabolic blood markers, and to improve body mass and composition. The modalities for treating humans and animals include oral administration to reduce the LDL/HDL ratio or the proportion of triglycerides/HDL, or topical administration as a cosmetic treatment to reduce fine lines, wrinkles, skin discolouration and other skin disorders.

Description

PERICARPIO DI CACAO COMPOSITIONS AND METHODS FOR

YOU WILL PRODUCE

Field of the Invention and Introduction

The invention relates to economically advantageous uses of the pericarp of the Theobroma cacao plant pod and edible products and compositions derived from this material. The pericarp of the cocoa pod is typically a residual product in the production of cocoa beans and the methods described here make use of what would once be discarded cocoa material. In particular, the invention employs the pericarp of the sheath and its commercial use in the production of a dietary supplement or food ingredient. Also, as shown here, they are method of the invention that result in compositions that unexpectedly possess high levels of beneficial compounds, which result in products and treatments to reduce blood triglyceride levels, to reduce LDL levels, to reduce blood glucose levels, for maintenance and weight loss, to reduce waist circumference, to improve muscle strength, and to improve other cardiometabolic markers in the blood. The other aspect, the pericarp material can be used to make a topical cosmetic composition, and in an example a topical cream, which can deal with coloration, wrinkles, and fine lines on the skin, and another skin condition. Background of the Invention

Theobroma cacao pericarp (CP) is an agricultural byproduct generated in abundance in plantations and farms when mature cocoa beans or seeds are removed in the pod processing. The term CP is used here because it is a specific term defined in plant biology and refers to the outer tissues of the cocoa pod. Cuatrecasas, J., cocoa and Its Allies: A Taxonomic Review of the Genus Theobroma, Contributions from the United States National Herbarium (Washington DC) 35, part 6. 1964. The pericardium surrounds the cocoa bean and the pulp of the surrounding seed . Other casual terms that can be found in the prior art may include the cocoa shell, the cocoa shell, and the cocoa pod, but these terms are confusing and frequently used to refer to cocoa bean structures, such as the shell of the seed. In the chocolate industry, the seed cover is part of the whole cocoa beans that have typically been fermented and dried. This seed material is not the pericardium. Seed material can often be referred to as a shell, cocoa bean shell, pod, shell or cover material in the scientific literature and in patent documents. The cover of the seed is removed from the grain of the seed either before or after roasting grain and before starting the manufacture of chocolate. Therefore, to avoid confusion, the term CP (pericarp of Theobroma cacao) in this invention.

CP is a main residual product of cocoa bean production on farms or plantations, and therefore for the cocoa bean production industry (Yapo, BM et al. Adding valué to cocoa pod husks as a potential antioxidant -dietary fiber source. Ana. J. Food Nutr. 1, 38-46. 2013 Karim et al., Phenolic composition, antioxidant, anti-wrinkles and tyrosinase inhibitory activities of cocoa pod extract. BMC complementary and alternative medicine, 14, 381 (2014) According to the International Cocoa Organization (ICCO), 4.4 million tons of cocoa beans were produced during 2013-2014, and approximately 3 million tons of CP woke up from farms and plantations, causing environmental problems, bad odors, and spread of diseases, such as black pod rot (ICCO 2015, Karim et al., 2014, Vriesmann et al., 2011, Vriesmann et al., 2012).

In Mexico and other cocoa-producing countries, processing the cocoa pericardium (CP) in useful by-products could provide economic advantages to farmers and reduce environmental problems (Vriesmann et al., Cocoa pod husks {Theobroma cacao L.): composition and hot-water-soluble pectins. Industrial Crops and Products, 34, 1173-1181 (2011). CP is rich in a variety of compounds, making it a promising source of such molecules; in this way, it could be a potential for production of useful natural compounds. The total phenolic content of the cocoa pod shell sheath is 46.4 ± 0.04 mg of gallic acid equivalents / g, which shows that it is a rich source of phenolic compounds that could be used instead of being discarded (Karim et al., 2014).

However, the preservation of the phenol content of the CP is challenging because this material decomposes rapidly, especially in hot, humid, humid tropical climates. In addition, the location of most cocoa plantations generally does not allow local storage or conservation methods. Fruits, vegetables and other perishable agroindustrial by-products can be dehydrated to preserve them for later use (Si et al., Comparison of different drying methods on the physical properties, bioactive. Compounds and antioxidant activity of raspberry powders. Journal of the Science of Food and Agriculture (2015) Dehydration prolongs the shelf life of the final food product, allowing its long-term storage and convenient future use in the manufacture of food, beverages or supplements, among the different drying processes used, it is commonly used Hot air drying in food production because it is an economical method, despite the fact that a long drying period can result in a lower quality product.

In addition, the use of high temperatures for Dehydration is controversial because naturally bioactive compounds are relatively unstable under heat and, in this way, nutrients can be lost during thermal processing (Choi et al., 2006). Conversely, studies have shown that the phytochemical content and antioxidant and biological activities of processed foods can be increased by heat treatment causing degradation of cellular constituents, thereby promoting the release of photochemicals in the environment (Choi et al., 2006, Wojdylo et al., 2009).

Although several methods and conditions for drying are presumed to differentially affect the quality and quantity of the bioactive compounds present in fruits and vegetables, there is no information regarding the effect of the use of drying techniques on the bioactive compounds present in the CP. Addition, nothing is known about the beneficial effect of such methods on the content of polyphenol or ^'antioxidant in these materials when consumed by humans.

The invention, in part, addresses these deficiencies and provides compositions produced from the CP material that can be used in human and animal consumption. As explained below, the methods of the invention can be used to produce a food supplement or a food ingredient which, when consumed regularly, advantageously leads to a dramatic reduction of blood triglyceride levels, and a beneficial improvement in both LDL and HDL levels. In addition, methods to treat subjects with the CP dietary supplement or ingredient results in weight loss and improvements in muscle strength. In another aspect, the CP material is used to make a topical cosmetic composition or a topical cream that can be used to treat discoloration, wrinkles, fine lines or other skin conditions.

Brief Description of the Invention

In one aspect, the invention provides a new source rich in beneficial cocoa-derived compounds, such as antioxidants, polyphenols, and flavonoids. In another aspect this invention provides methods for treating humans and animals to improve the levels of various blood markers associated with diseases and / or improved health. In another aspect, the invention encompasses commercially advantageous methods for producing a food supplement, ingredient, or functional food made at least in part from the pericarp material of the cocoa pod. The "pericarp" is distinguished by three layers: the exocarp or outer skin of the sheath; the thick white mesocarp just below the exocarp; and a thin translucent inner layer called endocarp (see Figure 1). Collectively these layers comprise the outer part of the fruit or pod of cocoa and collectively are referred to as pericardium. Cocoa beans or seeds and the outer layer or membrane surrounding the seeds is different from the pericardium. The seeds or grains are also surrounded in the pod by a mucilaginous pulp material, which, again, is different from the pericardium.

Cocoa nuggets, the part of the seeds used in making chocolate, are surrounded by the cocoa shell, and the cocoa nuggets and shell are not part of the pericarp.

In a more specific aspect, the invention includes a method for the preparation of a "flour" food composition ingredient of the CP. The method comprises isolating the pericardium material from the cocoa pods, where the pericardium material is free of cocoa beans and pulp, forming a paste and drying the pericardium paste, and then crushing the dry material into a type product. flour or processing it in another food ingredient, such as a powder, paste, or extract. In preferred embodiments, the method employs a freeze-drying step under vacuum pressure until the pericardium material has a humidity level of 10% or less, 8% or less. In another preferred embodiment, the method employs a microwave drying step until the pericardium material has a humidity level of 10% or less, or 8% or less, or 5% or less. The pericarp material can be milled to obtain a pericarp meal with a particle size of less than 0.500 mm, or less than 0.425 mm.

In any of the methods or uses of the Compositions described herein, the invention may include the measurement of the polyphenol content, the total flavonoid content, and / or the antioxidant capacity of the pericarp material or the flour or food ingredient composition. In this way, the invention allows the production of compositions and the monitoring of advantageously high beneficial levels of flavonoids and polyphenol is derived from cocoa in those compositions.

In another aspect, the invention encompasses methods for improving the anti-oxidation capacity of the cocoa bean shell material comprising the isolation of the pericarp material from the cocoa pods after harvesting the cocoa beans from the pods of such so that the pericarp material is substantially free of cocoa beans and pulp material, then dry the pericarp material at a humidity level of 10% or less, or 8% or less. The dried material is then crushed to form a flour composition or other food ingredient. The drying method can be selected from one or more of vacuum freeze drying, oven heating, and microwave treatment. The methods may further comprise measuring the activity of the polyphenol oxidase of the pericarp material or of the flour composition or the food ingredient.

In another aspect, the invention includes prophylactic methods and methods for treating a human subject and / or animals by providing a CP ingredient derived only from the pericarp of the sheath, and administering the ingredient daily or multiple times per day. For human subjects in particular, the severely optimally delivered amount will include sufficient of the pericarp-derived cocoa composition to deliver approximately 12.5 mg of the equivalent measured total cocoa flavonoids of (-) -epicatechin per dose. Other amounts, doses and administration regimens can also be used and adopted. In the embodiments of the invention related to methods for treating human animals, or methods for treating or preventing diseases in humans or animals, the antioxidant capacity of cocoa-derived material improves the physiological markers associated with certain disease conditions. For example, one of the following markers or measurements can be used: triglyceride levels; LDL levels; glucose levels, ratio of LDL / HDL, body mass index; muscle mass; or triglyceride / HDL ratio. Accordingly, daily treatments with beneficial cocoa-derived compounds, or cocoa pod shell pericarp flour described herein, can advantageously improve health or prevent disease.

In other aspects, the methods for preparing ingredients and compositions may result in a flour-like composition, a paste, and / or an extract of CP. Various extraction methods are known in the art and may include the use of chemicals and solvents approved for food. Administration of the ingredients or compositions can be done with any suitable carrier, in solid or liquid form such as tablets, capsules, powders, soft gels, solutions, suspensions, emulsions, ointments, or creams. The ingredient or composition can also be administered as a food supplement or as a functional food ingredient. An extract of the pericarp or flour material can be used alone or in combination with one or more other plant extracts to produce a composition optimized for a particular functional effect. In addition, an extract composition can be formulated with one or more other dietary nutrients, such as vitamins, minerals, amino acids, etc., to provide a functional or nutritional supplement for the desired health effect. These ingredients may be combined with necessary binders, excipients, preservatives, colors and the like known in the industry or commonly used or described in similar products in order to produce a tablet, capsule, pill, liquid, cream, powder, or suitable food ingredient. As indicated, some of its ingredients or compositions can also be used for topical treatments or used in cosmetic products.

In another aspect, the CP flour composition can be used in a topical cosmetic composition, and in a Example a topical cream. The various CP compositions described herein may be mixed with a cosmetically acceptable carrier, which may include purified water, oils, alcohols, glycols, other cosmetically acceptable components available, and combinations thereof. Topical cosmetic compositions and methods for using them employ CP flour present in a range of about 0.1% to 1% by weight, or 0.05% to 1% by weight, or 0.05% to 5% by weight, for example.

In yet another aspect, the topical cosmetic composition may also include additional ingredients such as penetration enhancers, humectants, lubricants, pharmaceutically active agents, color agents, fragrances, preservatives, antioxidants, chelators, neutralizers, amino acids, anti-inflammatory agents, anti-inflammatory agents. irritants, anti-adhesive agents, astringent, binders, catalysts, stabilizers, emollients, emulsifiers, surfactants, cell signaling agents, retinols, essential oils, plant / botanical extracts, conditioners, film formers, gel-forming agents, agents foaming agents, scrubs, vitamins, minerals, pH adjusters, proteins, peptides, tactile enhancers, saccharides, solvents or any combination thereof.

In even another aspect, the topical skin care composition can be formulated as a cream, lotion, serum, facial cleanser, tonic, eye cream. sun block, bar, spray, impregnated personal care device, impregnated wipes, gel, fluid / liquid, soap, oil, butter, scrubber, cleanser, mask, concentrate, or any other form known in the art.

In another aspect, the present invention includes a topical skin care composition comprising a CP flour composition and a cosmetically acceptable carrier together with one more of the following: (a) a source of bio-retinol (b) a Retinol-like activity source; (c) a source of sodium hyaluronic acid or other cosmetically active acid; (d) a source of a moisturizing saccharide complex; (e) a source of ascorbic acid; and (f) a source of antioxidant components.

In even another aspect, a topical cosmetic composition according to the principles of the invention or the following examples applies to the skin, such as to the hands or face, which may have, for example, but is not limited to wrinkles. , fine lines, an uneven tone, loss of firmness, rough surface, dark circuits, boleas under the eyes, sun damage, redness, dryness, irritation, elongated ports and combinations of all or some of the above. Alternatively, the topical cosmetic composition can be applied to the skin to prevent the occurrence of several of the problems described above. Thus, the invention includes methods for the treatment and prevention of skin conditions. using a CP flour composition in a cosmetically acceptable carrier. Such methods include the daily use of the topical cosmetic composition on the skin, and the use of multiple times during a day, for a period of time such as four weeks or more.

The invention is not limited to the foregoing, or by the following Figures, Description, Claims, or Examples, but can be seen in its entirety from the entire contents of this description. None of the references or information cited should be taken as limiting or modifying any specific description or definition as used in this description.

Description of the Figures

Figure 1 is a top view of a cross-sectional image of a Theobroma of cocoa pod showing the layers of plant matter and the relative vexation of the seeds, the exocarp, mesocarp, and endocarp (Cuatrecasas, J. 1964). The "pericarp" is distinguished by three layers; the exocarp or outer skin; the thick white mesocarp just below the exocarp; and a thin translucent inner layer called the endocarp. Collectively these layers comprise the outer part of the fruit and are of maternal origin. These seeds are surrounded by mucilaginous pulp material, and both the seeds and the pulp are not part of the pericarp.

Figure 2 is a schematic showing a side cross-sectional view of a cocoa pod Theobroma that It lists the location of the seeds, exocarp, mesocarp, and endocarp.

Figure 3 shows the total phenolic content (TPC) and the total flavonoid content (TFC) of fresh and dried CP flour prepared by different methods: OHD - oven-dried heat ; MWD - microwave dried; and FD-freeze drying. The total phenolic content is expressed as mg of gallic acid equivalent / 100 g of dry sample mass (DM) and the total flavonoid content is expressed as mg of equivalent of (-) -epicatechin / 100 g dry sample mass. The levels in the graphs with different letters are statistically significant (p <0.05).

Figure 4 shows the changes in grip strength after eight weeks of placebo use or experimental cookie treatment (cookies) (placebo cookies and cookies containing CP flour). There is a positive and significant increase in strength with the regular treatment of CP flour.

Figure 5 illustrates an example of the wrinkles observed on the back of a volunteer's hand before the application of the CP-based skin cream and after four weeks of local application.

Figure 6 summarizes the changes observed in the total height of wrinkles on the back of the hand observed in subjects exposed to four weeks of either a placebo or a skin cream based on CP. As you can tell A statistically significant reduction in the total height of wrinkles is noted in subjects treated with PC.

Figure 7 summarizes the observed changes in the color of the back of the hand noted in subjects exposed to four weeks of either a placebo or a CP-based skin cream. As can be seen, a statistically significant reduction in the color of the back of the hand is observed in subjects treated with PC.

Detailed Description And Examples

The invention in one aspect generally encompasses a method of producing and using a food ingredient of specific parts of the CP material. The ingredient, which can be formed as a flour for baking or food manufacturing, can be used to produce edible products and that promotes health, such as wafers, baked cookies, pastelee, and similar products. The ingredient can also be used as a supplement to be added to other food products or as a dietary supplement in the form of a pill or formulated.

As noted above, the preferred way to use the pericarp material is a flour-like composition that is capable of being used with a food ingredient or in a food product, such as a baked food. However, the invention is not limited to this preferred form or use. In particular, the compositions or extracts of the CP material of the invention can also be used as a nutritional supplement. In such form, the compositions can be introduced in a base carrier or on a carrier. The nutritional supplement can be included in a capsule, typically a water soluble capsule, but could be dispersible in water, thus releasing the nutritional supplement once it is mixed with water. A capsule could also comprise one or more of a gum, such as acacia gum and gum arabic, methylcellulose gum, dextrin, gelatin, caffeine, milk powder, skim milk powder, soy protein, vegetable protein, or other protein . Alternatively, or additionally, the food supplement may be carried in a granular, flake or particulate carrier, which could again be one or more of a gum, such as acacia gum, gum arabic, dextrin, methylcellulose gum, gelatin, soy, vegetable protein, legume seed protein, soy milk, soy milk powder, soy protein, caffeine, vegetable gum, cellulose, starch, sugar, glucose, and / or maltodextriña.

Advantageously, the dietary supplement that uses the CP material can be provided in a tablet form to be consumed once or several times per day. For example, in one embodiment, the dietary supplement comprises a portion of a single tablet, each serving suitable to be consumed one or three or more times a day, for example, in, before, or between foods as appropriate.

In alternative modalities, a Pharmaceutical composition containing at least one extract of the cocoa polyphenol of the pericarp material described herein, in an amount sufficient to obtain the desired effect on a subject, this composition may be a tablet, a liquid, capsules, soft capsules, paste , or pill, gum, or a drinkable solution or emulsion, a dry oral supplement, or even a wet oral supplement. The pharmaceutical composition may also contain carriers and excipients that are suitable for supplying the logically active compounds of cocoa, as well as other compounds of a different nature. The carrier / excipient class and its amount will depend on the nature of the cocoa extract or cocoa compounds or substance used and the desired mode of delivery and / or administration.

In addition, cosmetic or cosmeceutical compositions can be produced from the CP material described herein. A topical form may be a solution, emulsion, serum, skin and / or hair cleanser, body shampoo, body scrubber, soap bar, liquid soap, shampoo foam, deodorant, skin care preparation and / or hair, foam, mus, cream, lotion, ointment, balm, bar, gel, pump spray, spray spray, and combinations thereof. In one embodiment, the cosmeceutical compositions of this invention can be used in foams in personal care applications such as soap dish, shampoos, skin cleansers. oral and similar products. The estimation compositions in this way may also comprise cosmetically acceptable carriers to act as a diluent, dispersant or carrier of the CP ingredients or extracts. The carrier may be one selected to facilitate the distribution and absorption of cocoa-derived compounds when the composition is applied or administered. Vehicles other than or in addition to water may include emollients, solvents, humectants, thickeners, powders and liquid or solid perfumes.

The following examples and descriptions are illustrative only and should not be taken as limiting the scope of the invention.

Example 1: CP flour production

Step 1: Production of a CP paste

Whole cocoa pods were washed with soap, played with water, disinfected for 15 minutes with Citrus21 solution * (20 ml / 1 water, Integra Citrus, Mexico City, Mexico). The pods were opened, the seeds and pulp were removed, and only the pericarp of the husk was retained and crushed in a semi-industrial mixer (Crypto Peerless K55, USA) until a paste was obtained.

Faso 2: Methods for drying the CP paste One or more of three methods can be used to dry the CP paste. The samples were dried using:

A. CONVENTIONAL HOT AIR OVEN (CD). 150 g of pasta were dried at 60 ° C for 24 h (Lindberg Blue oven OV-484, USA).

B. DRYING BY FREEZING (FD). 150 g of paste were dried under vacuum pressure of 20 Pa for 24 h (Scientz, SC-10N, Ningbo, China).

C. DRYING IN MICROWAVE (MWD). 150 g of pasta were dried in microwave energy (595 Watts) for 11.5 min using a Panasonic inverter NN-SN968, 2450 MHz.

Drying was conducted until the paste samples reached a moisture level of 4-8%.

Step 3: Screening and grinding dry pasta to produce cocoa pericarp flour

The dried cocoa pericarp paste was finely ground using a Cunill luxo mill and sieved to obtain:

Fine pericarp flour with a particle size of ≤ 0.425 mm.

Thick pericarp flour with a particle size of> 0.425 mm.

Example 2: Characterization of pericarp flour samples

The CP paste has a moisture content of 87.4 ± 0.56%. The drying process reduces the water content to 4.6-7.8% (see Table 1). The results indicate that when the freeze-drying methodology is used, a lower moisture content is obtained compared to drying the conventional or microwave paste. The moisture content of pericarp flour showed statistical differences (p <0.05) between the three drying methods used.

Table 1

Humidity, pH and water activity (Aw) of dried pericarp flour samples

* p <0.05 vs. conventional, ** p <0.05 ve. microwave For pH determination, a suspension of lg of the sample was prepared in 10 ml of distilled water with continuous stirring for 2 min. The pH was determined using a pH meter (model pH 210, Hanna Instruments). The water activity of the pericarp flour samples was measured using the Hygropalm water activity meter at »1 (av-D10). Triplicate samples were measured at 23.6 ± 2 ° C.

Water activity (Aw) and the pH of dry pericarp flour are important parameters related to the deterioration and degradation of the product. The pH values were similar between the drying methods used, resulting in a pH in the range of 5.57 - 5.7 (Table 1). The activity of the dried pericarp flour gua had low values in the range of 0.252 (freeze drying) to 0.543 (microwave), showing differences statistics among them (p <0.05) (Table 1). Interestingly, despite the fact that the microwave drying method presented the highest humidity and values Aw, these are below the safe storage limit conditions that prevent enzymatic and microbiological deterioration, indicating that any of these three Drying methods can be used if only the conservation of CP flour is considered.

The color evaluation of these flour samples can be conducted at room temperature using a color reader (CR-10, Konica-Minolta Sensing Inc., Osaka, Japan), the measured parameters were L * for luminosity; C * for chrominance; A * for red; B * for yellow, the total color difference (ΔΕ) between fresh cocoa and dry pericarp flour was determined using an equation. ΔΕ indicates the magnitude of the color difference between the fresh and dry samples. These can be classified analytically as a slight difference (0.5> ΔΕ <1.5), notable difference (1.5> ΔΕ <3), marked difference (3> ΔΕ <6), extremely marked difference (6> ΔΕ <12), and one color with a different tone (ΔΕ> 12).

The color composition obtained from the three drying methods observed is presented in Table 2. Evaluation of flour color showed that the method of drying the cocoa pericarp flour used had significant effects, the main changes after drying were they give in Tone * and b * values, with an increase significant compared to the fresh pericarp sample which indicates the degradation of the original color by changing it to a more yellowish brownish color. The dried CP flour by means of the three different drying methods showed a large ΔΕ of the fresh pericarp sample (> 20), which indicates a total color tone change, which can be seen with the naked eye. Freeze drying and as a result the greater color change of the pericarp between the drying methods. The color results after drying treatments are important since the color changes could affect the sensory properties of the CP flour, limiting its use as a food ingredient, and also directly related to changes in the composition of the samples separated.

Table 2. CP flour color measurements obtained using different drying methods.

The color difference (ΔΕ) is calculated using fresh pericarp cocoa flour (Fresh) as a reference.

* = (p <0.05) fresh front.

The biochemical evaluation of flour extract from CP can use the determinations of the total phenolic or flavonoid content or the antioxidant activity of the method described by Martínez et al. (2012) with modifications. 500 mg of fresh and dried CP flours were stirred at room temperature for 60 minutes with 20 ml of acetone-water-acetic acid (70: 29.5: 0.5). These samples were centrifuged at 3,500 g, 15 min at 4 ° C and the supernatants were kept at -20 ° C until use.

Total phenolic content (TPC) was determined using the Folin-Ciocalteu reaction. A volume of 0.3 ml of the flour extracts was poured in triplicate into test tubes, adding 2.5 milliliters of Folin-Ciocalteu reagent (diluted 1:10 double distilled water) and 2 ml of sodium bicarbonate solution (7.5% p / v). The tubes were shaken and mixed rapidly using a vortex shaker, covered with for film and incubated at 50 ° C for five minutes. The absorption at 760 nm was measured with a Genesys 10 UV spectrophotometer. The concentration was determined using standard gallic acid curves and the results were expressed as milligrams of gallic acid equivalents (GAE) / g dry mass (mean ± SD).

The total polyphenol (TPC) content of the CP flour samples are shown in Figure 3. The poly phenolic content was different (p <0.05) between the three drying methods used. The highest concentration was found in freeze dried CP flour followed by microwave drying and the lowest by conventional oven drying. Freeze-drying probably caused the retention of the higher phenolic content since the drying method does not use heat to dry the samples, while with conventional oven drying, the oxidative and thermal declaration or interlacing of the polyphenols with pericarp structures probably increases given the long duration of heat treatment. Interestingly, the microwave drying process retained a considerable amount of phenolic compounds (a few milligrams below freeze drying) since it applies a short heating time, therefore, reducing thermal degradation.

The total flavonoid content (TFC) can be determined following the method described by Maleyki & Iemail, 2010. 1 ml of cocoa pericarp flour extract and 0.3 ml of NaNO ₂ (0.5%) was added. The mixture was reacted for five minutes after addition of 0.3 ml of AIC1 ₃ 6H2O (10%). The mixture was stirred and allowed to react for six minutes. 2 ml of 1 M NaOH was added and the total volume was adjusted to 10 ml with water. A UV spectrophotometer was used to read absorbance 510 no. TFC was calculated with a calibration curve using (-) - epicatechin and expressed as equivalents of (-) - epicatechin / g dry mass.

TFC results are also shown in Figure 3. Flavonoid content was different (p <0.05) among the three drying methods used. The highest concentration was found in freeze dried cocoa pericarp flour followed by microwave drying and the lowest by conventional oven. The microwave drying process retained a considerable amount of flavonoid.

The antioxidant activity can be determined by the radical scavenging properties of the flour samples, evaluated using the method previously reported by Brand-Williams et al, with some modifications. A stock solution was prepared by dissolving 24 mg of DPPH (2,2-diphenyl-l-picrylhydracil) with 100 ml of methanol and then storing at -20 ° C until necessary. The DPPH solution was obtained by mixing 10 ml of stock solution with 45 ml of methanol until an absorbance of 1.1 ± 0.02 units at 515 nm was obtained using the spectrophotometer. The CP flour extracts (0.150 ml) were allowed to react with 2.85 ml of the DPPH solution for 2 h in the dark. The absorbance at 515 nm was measured using a UV-VIS spectrophotometer. The radical DPPH radical scan rate of the samples was calculated using the following equation:

% Inhibition - [(Ajblanco - Ajnuestra) / A_blanco] x 100

A_blanco is the solvency of the solvent, and Ajnuestra is the absorbance of the samples of the samples. The standard curve was linear between 25 and 800 uN of the antioxidant Trolox synthetic. The results express in μΜ Trolox equivalent (TE) / g dry mass on average of the three replicates.

The results shown in Table 3 demonstrate that CP flour samples have the ability to sweep free radicals DPPH. Among the three treatments, the microwave and freeze drying methodology show a similar antioxidant potential, while conventional oven drying showed the lowest antioxidant capacity. These results suggest that phenolic content may play an important role in determining antioxidant activity. Some changes in polyphenols may appear during dehydration and could be related to the formation of the compounds or the formation of polyphenol derivatives with increased antioxidant activity.

CP flour exhibits a dose-dependent antioxidant activity according to the DPPH test (regardless of the method used to approach it) with a positive and significant correlation, freeze drying (R2-0.9786), conventional drying (R2-0.9561) and microwave drying (R2-0.99633).

For the ABTS test (2,2'-azino-bis (3- ethylbenzothiazolin-6-sulfonic acid)), the procedure followed the method described by Martínez et al., 2012. Stock solutions included 7.4 mM ABTS + solution and 2.6 mM of potassium persulfate solution. The operative solution It was then prepared by mixing the two stock solutions in equal amounts and allowing them to react for 12-16 hours at room temperature in the dark. The solution was then diluted by mixing 1 ml of ABTS + solution with 60 ml of methanol to obtain an absorbency of 1.1 ± 0.02 units at 734 nm. CP flour extracts (150 μΐ) were allowed to react with 2850 μΐ of ABTS + solution for 2 h in a dark condition and then absorbency was measured at 734 nm. The standard curve was linear between 25 and 800 µM of synthetic antioxidant Trolox. The results express in uM equivalent Trolox (TE) / g dry mass.

The results of the ABTS test are shown in Table 3. The drying methods generated a significant increase (p <0.05) in the antioxidant activity with freeze drying of the CP flours that produced the highest values. The highest antioxidant activity may be related to a better level of extraction of the antioxidant compounds from the pericarp matrix using this method.

Table 3. Antioxidant activity of fresh and dried CP flour measured by DPPH and ABTS methods.

In Table 3, OHD-drying by oven heating, MWD-drying in microwave, and FD-drying by freezing. Antioxidant activity was expressed as μΜ Trolox equivalent (TE) of dry mass (dm). * - (p <0.05) versus fresh.

Example 3: Clinical eight week clinical regimen using human subjects.

A placebo-controlled clinical trial can be used to use a baked biscuit food product to demonstrate the beneficial effects of pericarp meal consumed by humans and animals. Several clinical endpoints can be considered, including: body weight; body mass index (BHI); abdominal circumference; body fat; muscle mass; blood triglycerides (ΤΘ); Blood glucose; blood cholesterol; LDL cholesterol in blood; HDL cholesterol in blood; TG / HDL ratio; LDL / HDL ratio; and clamping force of the left hand.

Subjects were recruited under the following criteria: Men and women aged 20 to 60 years; BMI of> 25 kg / m ¹ at <29.9 kg / m ^a ; blood triglycerides 150 - 350 mg / dL. Subjects excluded by any of the following: type 2 diabetes mellitus; dyslipidemia; hypertension; cardiovascular diseases; subjects treated with hypolipidemic drugs, metformin or steroids; pregnancy or surgery in the last six months; lupus; arthritis; hypothyroidism; HIV or anti-retroviralee treatment; consumers of alcohol or tobacco; and a history of allergy to chocolate or cocoa components.

Two groups, placebo / control and experimental, were recruited. Both groups (placebo and experimental, n-20 subjects per group) received nutritional advice advising them to reduce 250 kcal / day from their usual diet and exercise moderately to burn another 250 kcal / day.

After random distribution of any group, the placebo group received control biscuits (~ 4 g and 14 Kcal) twice a day, once in the morning and joining in the afternoon, before meals. Subjects in the experimental group received cookies supplemented with CP flour (~ 4 g and 14 Kcal) containing 12.5 mg of total flavonoids measured in equivalents of (-) -epicatechin, twice a day, once in the morning and once in the afternoon , before food. Subjects, scientists and clinical laboratory technicians did not have access to treatment groups.

The baseline characteristics of the recruited subjects are shown in Table 4. Both groups were equivalent to the start of the eight-week trial.

Table 4. Baseline characteristics of the placebo and experimental groups.

The results of subjects treated with placebo and pericarp flour are shown in Tables 5 and 6.

Table 5. Anthropometric data after eight weeks of ingestion of cookies.

Table 6. Rate cardiometabolic endpoint in blood after eight weeks of ingestion of cookies.

The physical results after eight weeks of biscuit supplements are summarized in Table 5. Diet and exercise counseling induced favorable changes in both groups, but there are no differences between the groups. Changes in blood markers Cardiometabolics are summarized in Table 6. The consumption of placebo cookies did not induce significant changes in any of the markers evaluated. In contrast, the consumption of experimental cookies (containing cocoa pericarp flour) induced positive and significant changes in triglycerides, glucose and the ratio of TG / HDL. HDL was slightly reduced in the placebo group and showed a non-significant increase in the experimental group. These results indicate a reduction in cardiometabolic risk in subjects who consumed biscuits supplemented with CP flour. The experimental group (cocoa pericarp flour) shows evidence of a surprisingly significant reduction in blood triglyceride levels.

The holding force of the left hand (non-dominant hand) can be evaluated before and after placebo treatment or with cookies containing CP flour with a dynamometer (Camry Grip Strength Dynamometer, City of Industry, CA). Subjects were instructed to apply as much force as they could. The bow was repeated three times (30 seconds between tests), the highest value was recorded. Differences at the beginning and at the end of the study (eight weeks) were compared in both groups.

The subjects in the placebo group did not show any change in the strength of the hand grip. However, the ingestion of CP flour biscuits induces a significant increase in the strength of the hand holding (Figure 4).

Example 4:

The following pertains to the manufacture and use of topical cream using cocoa pericarp flour

(CP) and proof of its beneficial effects on human skin. For this study, a control cream (control cream) was manufactured and compared with the same cream in which CP flour (CP cream) was added. The basic cream ingredients are indicated in the following table:

Table 7. Components of the illustrative topical cream (CP cream)

Experimental setup

Analysis of the effects of the cream on the wrinkles of the hand and skin color. Subjects (n = 22, 45-65 of age) of both genders were randomly assigned to either a) control cream (n-10) or b) CP cream (n-12). The subjects were instructed to apply approximately 200 μΐ of cream each morning to the back of the right hand and spread it evenly over the hand each day for a period of four weeks.

For the analysis of the wrinkles of the hand, images were taken from the back of the hand using a mold to place the arm in the same uniform position while holding a 3 cm cylinder. Images were recorded using the Skin Analysis System

Multifunction (Skin Scope Analyzer EH-2100) at approximately the same point (middle part of the hand). The analysis of the acquired images was carried out using the envelope on the imaging computer J and the following wrinkle end points were generated: 1) total height of the wrinkle profile

(Rt), 2) highest peak (Rp) and, 3) lowest valley (Rv). Changes in hand wrinkles were analyzed by comparing Rt, Rp and Rv before and after four weeks of treatment using a comparable t test (statistically different if p <0.05). No statistical differences were detected in Rt, Rp and Rv with the control cream. As illustrated in Figure 5 and summarized in Figure 6 with the CP cream, significant reductions were detected in the lower valley (Rv) and, consequently, in the total height profile (Rt) of wrinkles (i.e., a flatter skin profile develops).

From the same acquired images, the color changes were analyzed using a different J image software option (3D color inspector plugin). As summarized in Figure 7, the CP cream worked to reduce skin pigmentation to a clearer overall intensity.

The terminology used in this document is intended to describe particular modalities only and is not intended to be limited to any method or approach, as the present document uses, the singular forms "a", "one, one", and "the, the "they intend to include plural forms as well, unless the context clearly indicates otherwise. In addition, it understands that the terms "comprises" and / or "that it comprises", when used in this specification, specify the presence of indicated characteristics, integers, steps, operations, elements, and / or components, but do not exclude the presence or adding one or more characteristics, integers, steps, operations, elements, components, and / or their groups.

The invention can be shaped in other specific ways without departing from the spirit or its essential characteristics. The present modalities are therefore considered in all aspects as illustrative and not restrictive, the scope of the invention is indicated by the claims of the application instead of by the above description, and all changes that fall within the meaning and equivalence range of the claims are therefore intended to be encompassed there.

It is noted that, in relation to this date, the best method known by the applicant to implement said invention is that which is clear from the present description of the invention.

Claims

CLAIMS Having described the invention as above, the contents of the following claims are claimed as property:

1. A method for the preparation of cocoa fruit flour, characterized in that it comprises isolating the pericarp material from the cocoa pods, between the pericarp material is free of cocoa seeds and pulp, grinding the pericarp material to a paste , freeze dry the paste under vacuum pressure to a level of less than 10%, and also crush the dried pasta to form a flour composition.

2. The method according to claim 1, characterized in that the drying is maintained until the humidity level of 8% or less.

3. The method according to claim 1, characterized in that the dried pulp is ground until the average particle size is 0.500 mm or less.

4. The method according to claim 2, characterized in that the paste is crushed until the particle size is 0.425 mm or less.

5. The method according to claim 1, characterized in that it further comprises measuring the total polyphenol content of the material of the pericarp or of the. flour composition

6. The method according to claim 1, characterized in that it further comprises measuring the total flavonoid content of the pericarp material or the flour composition.

7. The method according to claim 1, characterized in that it further comprises measuring the total antioxidant content of the pericarp material or the flour composition.

8. The method according to claim 1, characterized in that it further comprises using the flour composition to prepare a food product.

9. A method for the preparation of cocoa bean husk flour characterized in that it comprises insulating the pericarp material from the cocoa pods, where the pericarp material is free of cocoa beans and pulp, grinding the material of the pericarp to a paste, microwave the pasta to a moisture level of less than 10%, and then crush the dried pericarp material to form a flour composition.

10. The method according to claim 9, characterized in that the drying is maintained until the humidity level of 8% or less.

11. The method according to claim 9, characterized in that the dried pulp is ground until the particle size is 0.500 rom or less.

12. The method according to claim 9, characterized in that the dried pulp is ground until the particle size is 0.425 mm or less.

13. The method according to claim 9, characterized in that it further comprises measuring the total polyphenol content of the pericarp material or the flour composition.

14. The method according to claim 9, characterized in that it further comprises measuring the total flavonoid content of the pericarp material or the flour composition.

15. The method according to claim 9, characterized in that it further comprises measuring the total antioxidant content of the pericarp material or the flour composition.

16. The method according to claim 9, characterized in that it further comprises using the flour composition to prepare a food product.

17. A prophylactic method for treating a human subject characterized in that it comprises providing an ingredient in the shell of the cocoa pod derived only from the pericarp of the pod, and administering the ingredient daily at an amount that provides the total polyphenol content equivalent to approximately 12 mg of (-) -epicatechin per dose twice a day.

18. The method of conformity with the claim 17, characterized in that it further comprises monitoring one or more of the following cardiometabolic markers in the subject's blood: triglyceride levels; LDL levels; glucose levels; LDL / HDL ratio, body mass index, muscle mass, or triglyceride / HDL ratio.

19. A method for reducing blood triglyceride levels in a human subject with elevated glyceride levels, characterized in that it comprises preparing a composition comprising an effective dose of a cocoa pericarp flour or cocoa pericarp material extract, and orally administer the composition daily to the subject daily.

20. The method according to claim 17, characterized in that the subject has triglyceride levels between 150-350 mg / dL before treatment.

21. The method according to claim 17, characterized in that the composition is administered before a food.

22. The method according to claim 17, characterized in that the composition is administered during a food.

23. The method according to claim 17, characterized in that the composition is administered between foods.

24. The method of conformity with the claim 17, characterized in that the table composition daily before each food.

25. The method of using a CP flour made in accordance with the method of claim one, characterized in that it further comprises adding CP flour to foods, beverages or supplements to lower triglycerides in humans.

26. The method according to claim 25, characterized in that it further comprises adding CP flour to foods, beverages or supplements to reduce blood glucose in humans.

27. The method according to claim 25, characterized in that it further comprises adding CP flour to food, beverages or supplements to reduce the proportion of LDL / HDL in humans.

28. The method according to claim 25, characterized in that it further comprises adding CP flour to food, beverages or supplements to reduce the proportion of glycerides / HDL in humans.

29. The method according to claim 25, characterized in that it further comprises adding CP flour to food, beverages or supplements to reduce waist circumference in humans.

30. The method according to claim 25, characterized in that it further comprises adding CP flour to food, beverages or supplements to reduce the body mass index in humans.

31. The method according to claim 25, characterized in that it further comprises adding CP flour to food, beverages or supplements to increase muscle mass in humans

32. The method according to claim 25, characterized in that it further comprises adding CP flour to food, beverages or supplements to increase the strength of muscle support in humans.

33. A topical cosmetic composition characterized by porgue comprises a CP flour composition and a cosmetically acceptable carrier.

34. The composition according to claim 33, characterized in that it further comprises one or more of: (a) a source of bio-retinol (b) a source of retinol-like activity; (c) a source of sodium hyaluronic acid or other cosmetically active acid; (d) a source of a moisturizing saccharide complex; (e) a source of ascorbic acid; and (f) a source of antioxidant components.

35. A method of treating the skin through topical administration of a CP flour characterized in that it comprises providing a composition according to claim 33 and applying it to the skin of a patient at least once a day.

36. The method according to claim 35, characterized in that the CP flour is present in the composition at a range of 0.1% to 1% by weight.