IT201800007431A1 - LIQUEUR WITH HIGH POLYPHENOL CONTENT AND PROCESS FOR ITS PRODUCTION - Google Patents

Description

2. DESCRIPTION

of the invention with TITLE: "Liquor with a high polyphenol content and process for its production"

2. 1 BACKGROUND OF THE INVENTION

The industry of food supplements in fluid form, such as herbal extracts, has had a considerable development in recent years, such as to attract the attention of even the most well-known and important pharmaceutical companies.

In particular, among these, for several decades Propolis has been the subject of pre-clinical studies, in vitro and in vivo. Regarding its chemical composition it must be said that in recent years science has made considerable progress in elucidating the chemical composition of Propolis (Huang S, Zhang CP, Wang K, Li GQ, Hu FL; Molecules. 2014 Nov 26: 19 (12): 19610-32), and despite the undoubted variability of this, due to the diversity of the environmental context of the various geographical areas of origin, today the effectiveness of propolis understood in a general sense, at least in improving the conditions of inflammation , as well as infection of various kinds, of the human oral mucosa (and not only) is amply demonstrated and confirmed by recent bed bug studies (Pereira EM, da Silva JL, Silva FF, De Luca MP, Fermila EF, Lorentz TC, Santos VR; Evid Based Compìement Alterna † Med.

2011: 750249, Jain S., Rai R., Sharma V., Batra M. World Journal of Pharmaceutìcal Sciences. 2G14; 2 (l); 90-94. Bertolini PF, Biondi Filho O, Pomilio À, Pinheiro SL, Carvalho MS; J Appl Oral Sci. 2012 Feb; 20 (1): 32-7. Skaba D, Morawiec T, Tanasiewicz M, Mertas A, Bobela E, Szliszka E, Skucha-Nowak M, Dawiec M, Yamamoto R, Ishiai S, Malata Y, Redzynia M, Janoszka B, Niedzielska I, Król W Evid Based Complement Alternal Med. 2013: 215391.

The aforementioned scientific investigations go hand in hand with the massive release on the market of a large quantity of propolis-based products, registered as supplements, but also as medical devices, mostly in the form of hydroalcoholic extract, which is the one that has the most feedback on the level of effectiveness; the presence of ethyl alcohol is in fact essential, to allow both the dissolution of the raw propolis with consequent extraction of its active ingredients, and an effective local diffusion of them in the mucous membranes.

Today, more or less concentrated fluid formulations based on propolis extracts are found on the shelves of herbalists, pharmacies and parapharmacies, and in the health sector of many hypermarkets. However, a major limitation to the consumption of this form of product is the very unpleasant taste of these hydroalcoholic solutions. In fact, it is no coincidence that the solutions considered most effective are those with a higher alcohol content, precisely because they are able to solubilize a greater quantity of propolis and therefore also boast a higher content of flavonoids (the polyphenols of propolis), such as for example the pure hydroalcoholic extract of the or that.

But in this way, as is well known, a series of less noble compounds are also solubilized, and moreover with a dark color and an unpleasant taste, elements that characterize the typical concentrated alcoholic solutions of propolis present on the market, wrong that the aforementioned products have an extremely limited response from the potentially interested catchment areas. Furthermore, due to the particularly reactive characteristics of a solvent such as ethanol, extractive processes that require solutions with a high alcohol content, especially if hot, subject the natural substances present to a condition of stress that is detrimental to their stability with partial compromise. of their native structure; oxidative stress on polyphenols which degrade to quinones is a classic example.

Some only partial solutions to the aforementioned problem of the right compromise between taste and efficacy can be found on the market: products such as Larix (Laboratori Larix, Verona; it is a 10% extract of propolis in an alcoholic solution of 75% vol.) addition of fruit juices, with the aim of sweetening the flavor; but the total polyphenol content is mostly unknown, and the alcohol content is still high. On the other hand, other companies such as Cosval and Specchiasol, use a Propolis, subjected to industrial refining processes, in compositions containing only small quantities of alcohol and with the addition of multiple aromas, thus moving away from the more standard conditions of use. documented in the literature in terms of efficacy.

In any case, all the concentrated alcoholic preparations based on propolis do not solve the problem of the stability of the polyphenols present, of their tendency to oxidize even in a short time, as can be verified by a careful analysis of the very dark oxidative degradation pigments that characterize such concentrated alcoholic solutions, similarly to what happens in the degradation of polyphenols in wines, if they are not treated with suitable preservatives, such as bisulfites.

Therefore, the game of finding a compromise between a high concentration of active ingredients of a natural and native form of propolis and a formulation that marries the consent of consumers with an alcohol content that possibly does not exceed 50% remains open.

The nature and origin of the ethyl alcohol used for the preparation of tinctures and infusions of this type could make the difference to help solve the problem, as small quantities of substances contained in the same (impurities) can affect both the composition and on the organoleptic properties of the final extract obtained, as well as on its stability and conservation.

The distillates obtained from marc (or of other biological nature) commonly used for the preparation of grappa, vodka, or other liqueurs are an example of a sustainable and biological alternative to the use of industrial ethanol, but these liqueurs are nothing more than hydroalcoholic solutions, which they have the further advantage of containing a whole series of natural organic substances, including antioxidants, whose chemical structure and quantity is on the other hand widely documented today (see for example “Panighel, A., Ponchia, G .; Study of the Influence of Different Storage Conditions of Pomace on Varietal and Fermentation Aromatic Compounds - PhD Thesis in Viticulture and Oenology, Cycle XXI - Dept. of Agricultural Biotechnology, University of Padua).

Well, surprisingly, there is still no liqueur that boasts the presence of a propolis extract; similarly, there is no hydroalcoholic extract of propolis in which the hydroalcoholic mixture used is in fact constituted by a brandy. Yet the composition of the aforesaid spirits is not at all incompatible with that of a classic hydroalcoholic extract of propolis of the same alcoholic degree; indeed, it is certainly noteworthy that most liqueurs already contain polyphenols of various kinds and concentrations, depending on the origin of the fermented sugar matter and the type of manufacturing process used (Erik L. Regalarlo et al, Journal of Chromatography A, 2011, 7358-7364). More generally, it must be said that there are currently no liquid herbal liquor-based extracts on the market, except those for which traditional consumption as an alcoholic beverage is envisaged, when instead today the advanced and updated titration and standardization processes they can allow us to use simple and compound liqueurs for purposes of greater scientific rigor, including those inherent in the sphere of curative products, or at least food supplements.

In support of this. it should be emphasized that technical / scientific applications. therefore different from their consumption as liqueurs, of the aforementioned distillates, it is already a reality: in a recent study (ACS Sustainable Chem. Eng. 2015) a surprising replacement of the chromatographic eluents for HPLC with suitable hydroalcoholic mixtures constituted precisely by the distillates is successfully carried out in question, effectively opening the doors, even for high-level scientific applications, to these hydroalcoholic artifacts of natural origin.

2.2 DETAILED DESCRIPTION OF THE INVENTION

By investigating the concentrations of natural active substances in Propolis extracts, in particular of the total polyphenols present in hydroalcoholic fluid extracts of this raw material, both the concentration in simple hydroalcoholic extract of these substances and the concentrations obtained with a similar preparation were analyzed, but starting from pomace distillate having the same alcohol content. Unexpectedly, the extracts prepared with distillate proved to be more concentrated, in terms of the content of the aforementioned substances, than those prepared with a mere mixture of commercial ethyl alcohol and water. The same result was obtained with extracts prepared with Vodka (grain distillate) always identical alcoholic strength. From the dosages carried out, it appears that this greater quantity of the polyphenols in question is partly explained by their simultaneous presence in the distillates themselves used for the preparations.

A possible contributing cause of the greater quantity of flavonoids found in the liquor-based infusion, compared to the classic hydroalcoholic based one obtained at the same alcoholic strength, can be found in the influence that the set of additional natural substances contained in the spirits, such as terpenols and esters of various kinds, phenols, which may have a greater solubilization of the aforementioned polyphenols.

Starting from this experimental evidence, and from the need, appreciated by the general public, to use a resource that is closer to the food sector, and that marries better in terms of taste with the hardly acceptable taste of Propolis, we arrive at the object of the present invention , consisting of an infusion of raw propolis in which the components and the manufacturing process have peculiar characteristics such as to achieve said objectives of high quantity and stability of the polyphenols and greater satisfaction of the final user.

The infusion must be made using a quantity of raw propolis in particularly fine powder which is macerated inside the extractive fluid; the pulverization process can be facilitated by preventive freezing of the raw propolis batch, and the maceration is more effective by immediately dispersing the powder thus obtained inside the aforesaid fluid. The distillate used as a fluid can be good taste alcohol. Cognac, Grappa, Vodka or any other brandy, or a combination of them. The compound liqueur resulting from the provisions and reported in the following invention, in addition to being made up of a raw propolis spirit extract, optionally contains ediculants and one or more of one of the following additional ingredients:

1. one or more natural poiphenols from another herbal substance, or natural-identical or of a synthetic nature but identical to the natural counterparts.

2. herbal extracts in fluid, dry or soft form; non-exhaustive examples are those of echinacea, acerola, dog rose, licorice, star anise

3. essential oils, only the type and quantity for which internal use is foreseen. Non-exhaustive examples are those of Blue Chamomile, Roman Chamomile, Eucalyptus, Peppermint. Cloves.

It should be emphasized that these additional food ingredients can be an integral part of the starting raw material liquor or can be added downstream of the extraction process, forming the final product liquor.

By “raw propolis” we mean a resinous, balsamic material, rich in polyphenols, produced by bees, well known to experts in the beekeeping sector; these insects generally belong to the Apidae family, preferably of the genus Apis. Particularly suitable for the purposes of the present invention is Apis melìifera, which is precisely the honey bee, comprising various subspecies {Apis melìifera ssp.), Such as Apis mellifera mellifera, Apis mellifera ligustica, Sicilian Apis mellifera, Caucasian Apis mellifera . Propolis is characterized by a color ranging from yellow to dark green, sometimes brown, and comes from vegetable matter collected by bees on the shoots of some plants, such as exudates from buds, particularly of the Populus species; once this material has been collected, the bees process it, enrich it with wax and salivary secretions, and apply a layer of it on the inner surface of the hive, to close cracks and holes, to repair the cells, reinforce their edges and make them aseptic.

By Polyphenols we mean a vast class of organic compounds of vegetable origin known to experts in the sector; and quantitatively analyzed by means of multiple techniques - including the Folin-Ciocalteau method (redox), having structures with several hydroxylated aromatic rings, or more hydroxyls on the same ring, in free or ether form. Based on the scientific studies reported in the literature, the compounds contained in Propolis, in variable quantities with the geographical origin, include a considerable variety of natural active ingredients that fall within the class of polyphenols, such as benzoic acids, cinnamic acids and their derivatives (for example the esters of Caffeic Acid) or flavonoids, or compounds characterized by three hexatomic rings, 2 of which welded together, including flavones (such as Apigenin and Pinocembrin), isoflavones, flavanones (such as Chrism) and flavonols (such as Galangina and Quercetina).

Alcoholic preparations are understood to mean all preparations that come from any process that uses or produces a fluid with a high alcohol content of ethyl alcohol, including pure ethyl alcohol (good taste, 95% vol.).

The alcohol content of the extractive fluid of the present invention is between 15 ° and 95 °, preferably between 30 ° and 55 °, even more particularly between 35 ° C and 45 ° C. The alcohol content of the final produced liquor is between 15 ° C and 70 ° C, preferably between 30 ° and 55 °, even more particularly between 35 ° C and 45 ° C. Optionally, one or more phases of the process can comprise mixing, maceration, infusions, with flavors, herbs, fruits, concentrated or diluted fruit juices, extracted; for the aforementioned process, fluids particularly suitable for the purposes of the invention are alcoholic distillates from fermented vegetable raw materials, such as cereals, fruit, pomace. As non-exhaustive examples of such spirits and liqueurs we mention grappa, vodka, gin, rum, whiskey, cognac.

By aromas we mean those natural, non-natural or natural-identical substances, which serve to give a characteristic note to the flavor that fall within the scope of the provisions of Legislative Decree 25 January 1992, n. 107. Typical but not exhaustive examples of added aromas are the aromas of berries, peaches, sage, rose, rosemary, mint, cloves, anise, cinnamon.

By essences we mean alcoholic solutions of essential oils, such as lavender essence, or alcoholic solutions of chemical molecules used as flavorings for foods or cosmetics, such as thyme essence or vanilla essence.

By fluid or dry extracts we mean any natural extract, including the extracts of any herbal material that is included among the "botanicals" referred to in the "compendium of herbal substances and preparations" published by the European EFSA, as known to experts in the art. Typical but not exhaustive examples are extracts of licorice, juniper berries, rose hips, echinacea, aceraia, star anise, chamomile, ginseng, or other medicinal herbs.

By essential oils, as known to those skilled in the art, we mean any oily substance, or mixture of oily substances, obtainable by pressing, or by steam distillation of officinal herbs. Typical but not exhaustive examples are the essential oils of lavender, eucalyptus, chamomile, basil, jasmine, iris.

The liqueurs produced in the context of the present invention improve the concentration of flavonoids with respect to existing liqueurs, including those which here have the function of raw material. But at the same time, as hydroalcoholic extracts of Propolis, they maximize this concentration by minimizing the alcoholic strength, also improving the pleasantness of the taste, and contributing to a more natural and sustainable use of the alcoholic raw materials necessary for the manufacture of hydroalcoholic Propolis.

For the specific purposes of the present invention it is important to specify the physical and chemical characteristics of the final product liquor obtained within the present invention.

From the chemical / physical point of view, the final product has the characteristics of a liqueur composed of the above grade, which naturally contains high concentrations of polyphenols, in addition to the aforementioned optional added substances, such as sweeteners, extracts, vitamins, aromas, oils. essentials, dyes.

Its physical form is that of a light suspension, in which the liquid is of a color ranging from canary yellow to orange yellow, and the suspended solid tends to be white. Generally it tends to decant with the passing of the hours, even if this condition is not absolutely limiting of the product object of the invention.

From a purely chemical point of view, both qualitative and quantitative, the extract object of the present invention differs considerably from any other hydroalcoholic extract of propolis thanks to the following characteristics:

a) the relative and absolute quantity of polyphenols is different from what would be obtained by a similar infusion with a simple hydroalcoholic mixture of the same alcoholic strength. In particular, the chemical analysis shows a polyphenol content higher than that shown by a mere hydroalcoholic solution of the same strength available on the market, consistently with the additional presence of polyphenols in the white precipitate and, even if in a low percentage, with the presence addition of polyphenols naturally present in the liquor raw material.

b) this additional presence of polyphenols in the bottom body referred to in point a) implies that the quantity of polyphenols is such as to completely saturate the liquor itself for one or more polyphenolic rooms, as is evident from the very presence of the aforementioned bottom body , the typical precipitate of a lighter color than the mother liquors that those skilled in the art know to be the consequence of a precipitation consequent precisely to a saturation; the total quantity of polyphenols contained in this suspension, determined according to the Folin-Ciocalteau method of analysis, is between 1 mg / ml and 100 mg / ml, preferably between 2.5 mg / ml and 50 mg / ml and even more in particularly between 5 mg / ml and 25 mg / ml.

The total quantity of extracted matter is also typical of the final produced liquor in question. It can be determined by measuring the dry residue obtained after complete evaporation of the hydroalcoholic solvent, which is between 1% and 20%, preferably t and 2% and 10%

As mentioned above, in the scientific literature it is possible to find innumerable pre-clinical and clinical studies on the chemicals described above contained in Propolis, which not only confirmed what has been handed down by popular traditions on the beneficial abilities of Propolis, but have gone far beyond, demonstrating that the pool of polyphenols contained in Propolis act in synergy, exerting various pharmacological properties, namely antibacterial, antiviral, antifungal, immunostimulating, antioxidant and even anti-tumor properties. Since they are almost always polyphenolic fractions of propolis obtained by appropriate hydroalcoholic extraction of raw Propolis, the present invention is part of this context, creating a new class of products based on Propolis, characterized by being an extract in the form of liqueur having one or more elements of the aforementioned polyphenol pool in such quantity as to completely saturate the extract itself, as specified in b).

In a particular embodiment of the present invention, the alcoholic preparation in question possesses at least one of the aforementioned pharmacological properties, depending on the relative quantity (intended as a profile of the relative quantities of the individual species expressed as a percentage of the total) and therefore on the specific synergy of the aforementioned polyphenols present there. This relative quantity depends first of all on the geographical origin and therefore on the characteristics of the habitat of origin and can be modified, again within the scope of the present invention, with the addition of a further quantity of one or more natural-identical active ingredients with respect to what has already been mentioned above. present in the liquor produced, until the target pharmacological activity is maximized.

The production process used to obtain the aforementioned hydroalcoholic Propolis is typical for this type of manufacturing and includes any method known to experts in the field, such as to obtain a fluid extract, concentrated in its characterizing components, starting from an extractive liquid and a drug. (herb containing the natural active ingredients to be extracted) in powder form. Moreover, its description also constitutes for the experts an explanation of the aforesaid saturation, with the presence of a background body, of one or more polyphenolic elements. By way of non-exhaustive example, we report a process characterized by the following phases:

1. Introduction of the raw Propolis in powder form, inside the hydroalcoholic fluid having the above characteristics in the definition of alcoholic preparation, in an amount between 1% and 30% by weight, preferably between 3% and 20%, and more particularly from 5% to 15% with respect to this extracting alcoholic preparation.

2. Infusion at room temperature for a period between 3 hours and 30 days, preferably between 3 hours and 7 days, more particularly between 3 hours and 2 days

3. Heating of the aforementioned mixture at a temperature between 30 ° C and 80 ° C, preferably between 40 ° C and 70 ° C and more particularly between 45 ° C and 60 ° C for a period of time included between 10 minutes and 3 hours, preferably between 20 minutes and 2 hours, and more particularly between 30 minutes and 60 minutes.

4. Hot filtration of the obtained mixture; the yellow / orange filtered liquid, initially clear, becomes latigy. A first filtered fluid extract in the form of a light suspension is thus obtained.

5. Preferably, second infusion of the solid with an equal aliquot of alcoholic preparation of the first extraction, with the conditions described in point 2), followed by new filtration; this second filtered fluid extract thus obtained is combined with the first.

6. Optionally, third infusion, with repetition of point 4), and possible addition of water in order to adjust the alcohol content up to the desired alcohol content.

7. Optionally, addition of sweeteners, aromas, essences, fluid or dry extracts, essential oils, as defined above, thus arriving at the final fluid extract of propolis.

As an integral part of the present invention there is the use of the alcoholic fluid extracts thus obtained as Food Supplements, as Cosmetics and as Medical Devices, optionally inside vials equipped with a suitable spray or nebilization dispenser, in order to appropriately direct the jet of the infusion towards painful parts of the orophageal cavity, in case of inflammations such as gingivitis, pharyngitis, stomatitis, thrush.

2.3 EXAMPLES

Example 1

A batch of impure Propolis (1.5 kg) obtained from scraping of the anias (Apicoltura R Rohrwacher, Castelgiorgio -TR) is frozen at -20 ° C, pulverized with a professional bench coffee grinder (CEG, type MCMMP) and immediately placed always at -20 ° C. 100g of this powder are subjected to points 1) to 5) (including extremes) of the extraction process described above. In particular, 2 portions of 500 ml of marc spirit (grappa, alcohol 40% vol.) Are used, leaving the infusion to macerate for 10 days for both portions. at about 25 ° C; the heating phase of both infusions was carried out at 60 ° C for 1 h. The two portions are combined in a single batch of milky hydroalcoholic fluid extract, tending to the very slow formation of a white precipitate. The suspension is subjected to analysis of total polyphenols with the Folin-Ciocalteau method. B value obtained is equal to 11.4 mg / ml.

Example 2

From the same batch of Example 1 another 100g of pulverized propolis are taken, which is subjected to the same extraction process, with the difference that the distillate used in this case is cereal brandy (Vodka, alcohol 40% vol.). In this case the value of the total polyphenols obtained is equal to 11.7 mg / ml).

Comparative Example 3

From the same batch of Example 1 another 100g of pulverized propolis are taken, which is infused with a 40% vol. Hydroalcoholic mixture. of alcohol, prepared using pure alcohol (Alcol Buongusto 96 ° C). A 1 liter portion of this extracting mixture is placed in contact with the pulverized propolis for 20 days at 25 ° C. Once filtered, the resulting yellow / orange fluid remains clear without causing a precipitate. In this case the value of the total polyphenols obtained is equal to 8.2 mg / ml), about 30% less than the compound liqueur object of the invention.

Comparative Example 4

The Vinpepato liqueur, being the result of a concentrated extraction of a multitude of herbs, is one of the richest liqueurs in polyphenols among those on the market. Subjected to analysis of total polyphenols with the Folin-Ciocalteau method, it returned a value equal to 2.2 mg / ml and therefore approximately 5 times lower than the compound liqueur object of the present invention.

Example 5

The suspension obtained with the process referred to in Example 1 is placed in glass vials equipped with a pump and a pharmaceutical-type spray dispenser (Eurovetrocap) and distributed to a sample of 50 people who complained of sore throats or other inflammatory symptoms of the oropharyngeal cavity. , residing in the municipalities of Bagnoregio and Montefiascone (Viterbo). The individuals in question sprayed this suspension on the painful parts, 3 sprays 3-6 times a day, depending on the extent and persistence of the pain. After 5 days of treatment 46 of the 50 individuals declared the almost total disappearance of painful symptoms and redness of the mucous membranes.

Claims (6)

CLAIMS 1) A hydroalcoholic fluid extract of propolis supersaturated with polyphenols to the point of taking the form of a suspension, consisting of: a) Organic material identifiable as Propolis extract. b) Alcoholic distillates of fermentative vegetable origin including liqueurs produced with them. c) Optionally, Additional Ingredients consisting of sweeteners and at least one of: i) one or more natural or natural-identical polyphenols originating from herbal material other than propolis; ii) herbal extracts in fluid, dry or soft form including but not limited to those of Echinacea, Acerola, Rosa Canina, Licorice, Star Anise: iii) essential oils for internal use including but not limited to those of Blue Chamomile, Roman Chamomile, Eucalyptus Globulus, Peppermint, Cloves. 2) A propolis liqueur, consisting of an extract as defined in 1), having the following characteristics: a) alcohol content between 15% and 70%, preferably between 30% and 55%, more particularly between 35% and 45% (percentage by volume). b) content in polyphenols, determined by the Folin-Ciocalteau method, between 1 mg / ml and 100 mg / ml, preferably between 2.5 mg / ml and 50 mg / ml and more particularly between 5 mg / ml and 25 mg / ml. c) A solid residue, obtainable by complete evaporation of the hydroalcoholic solvent at 50 ° C and 20 mmHg, comprised between 1% and 20%, preferably between. 2% and 10%, more particularly between 3% and 7%. 3) A liqueur manufacturing process corresponding to point 2), a process consisting of: step a) Alcoholic infusion at room temperature for a period between 3 hours and 30 days, preferably between 3 hours and 7 days, more particularly between 3 hours and 2 days, to be made with i) hydroalcoholic mixture referred to in point 1) b) having an alcoholic strength between 15% and 95%, preferably between 30% and 55% and more particularly between 35% and 45% (percentage by volume) ii) finely pulverized crude propolis in an amount ranging from 1% to 30% by weight, preferably from 5% to 25% and more particularly from 10% to 20% with respect to the total hydroalcoholic infusion mixture; phase b) heating the aforementioned infusion at a temperature between 30 ° C and 80 ° C, preferably between 40 ° C and 70 ° C and more particularly between 45 ° C and 60 ° C for a period of time comprised between 10 minutes and 3 hours, preferably between 20 minutes and 2 hours, more particularly between 30 minutes and 60 minutes; step c) hot filtration of the resulting mixture; step d) preferably, carrying out a second extraction on the solid obtained by repeating steps a), b) and c) with an equal aliquot of the extracting hydroalcoholic fluid of the first infusion, followed by joining the two filtered liquids; step e) optionally, third extraction by repeating step d), using a different hydroalcoholic fluid. 4) A manufacturing process referred to in point 3) in which the optional ingredients referred to in point 1) c) are added at any moment of the process. 5) A hydroalcoholic extract as per the preceding claims 1), 2) and 3), optionally as per claim 4), packaged in any form of oral dispensation. 6) A hydroalcoholic extract for oral use as per claim 5) having at least one of the following properties: a) antiseptic properties, b) immunostimulating properties. c) anti-inflammatory properties.