Tincture For Infusing Flavonoids And Methods Of Use
20180000748 · 2018-01-04
Inventors
Cpc classification
A23V2002/00
HUMAN NECESSITIES
A61K8/498
HUMAN NECESSITIES
A23L33/105
HUMAN NECESSITIES
A61K47/10
HUMAN NECESSITIES
A23V2002/00
HUMAN NECESSITIES
A61K9/0095
HUMAN NECESSITIES
A61K2300/00
HUMAN NECESSITIES
A61K2300/00
HUMAN NECESSITIES
A61K31/00
HUMAN NECESSITIES
International classification
A61K31/00
HUMAN NECESSITIES
A23L33/105
HUMAN NECESSITIES
A61K47/10
HUMAN NECESSITIES
A61K9/00
HUMAN NECESSITIES
Abstract
Methods and formulations for batch-mixing flavonoid tinctures and infusing the flavonoid batch-mixed tincture into a food, beverage, cosmetic or drug products with minimal or no effect on the alcohol by volume of the product, and to precisely deliver the desired mixture concentration of the flavonoids, is provided. A batch-mixed flavonoid tincture includes an amount of at least two individual flavonoid tinctures, each flavonoid tincture being a mixture of at least a flavonoid and a solvent. The batch-mixed tincture is added to various food and beverages, or alternatively directly administered to a human or animal, to make a bioavailable amount of flavonoids to be absorbed and metabolized by the body providing demonstrated health benefits.
Claims
1. A method of making batch mixture of specifically desired combinations of bioavailable flavonoids, said batch mixture used for infusing a beverage, food, cosmetic or drug to provide a bioavailable dose of one or more flavonoids, the method comprising: making two or more flavonoid tinctures, each flavonoid tincture made by mixing an amount of flavonoid with an amount of solvent and agitating the flavonoid and solvent mixture until said flavonoid is dissolved in said solvent; and mixing an amount of two or more flavonoid tinctures to form a batch-mixed tincture solution.
2. The method of claim 1 wherein each flavonoid tincture has a different flavonoid than the others.
3. The method of claim 1 wherein each flavonoid tincture has a different solvent than the others.
4. The method of claim 1 further comprising filtering the batch-mixed tincture solution to remove any undissolved flavonoid, any impurities, any other suspended solids, or combinations thereof.
5. The method of claim 1 wherein the solvents is ethanol, dimethyl sulfoxide (DMSO), 1-butanol [n-butanol], 1-propanol, cyclodextrins, or combinations thereof.
6. The method of claim 1 wherein the flavonoid concentration of the flavonoid tinctures is between 0.001 and 50 grams per liter of solvent.
7. The method of claim 1, wherein the flavonoid is resveratrol, pterostilbene, quercetin, procyanidin, curcumin, or combinations thereof.
8. The method of claim 1 further comprising delivering the batch-mixed tincture into a body to a human or animal body intravenously, subcutaneously, intradermally, intramuscularly, sublingually, through the use of parenterally delivered products or combinations thereof.
9. A method of infusing a batch-mixed tincture, the method comprising: making two or more flavonoid tinctures, each flavonoid tincture made by mixing an amount of a flavonoid with an amount of a solvent and agitating the flavonoid and solvent mixture until said flavonoid is dissolved in said solvent for each flavonoid tincture; mixing an amount of two or more flavonoid tinctures to form a batch-mixed tincture solution; and adding an amount of the batch-mixed tincture solution to a food, beverage, cosmetic or drug product.
10. The method of claim 9 further comprising allowing sufficient time for the batch-mixed tincture to infuse with the food, beverage, cosmetic or drug product.
11. The method of claim 9 wherein said solvent is ethanol, dimethyl sulfoxide (DMSO), 1-butanol [n-butanol], 1-propanol, cyclodextrins, or combinations thereof.
12. The method of claim 9 wherein said flavonoid is resveratrol, pterostilbene, quercetin, procyanidin, curcumin, or combinations thereof.
13. The method of claim 9 wherein the solvent is ethanol and the amount of tincture changes the amount of alcohol by volume (ABV) of the food, beverage, cosmetic or drug product between 0.01% and 1.0%.
14. The method of claim 9 wherein the solvent is ethanol and the amount of tincture changes the ABV of the food, beverage, cosmetic or drug product between 0.02% and 0.5%.
15. A batch-mixed tincture for infusing a food, beverage, cosmetic or drug product to provide a bioavailable dose of flavonoid, the tincture comprising: an amount of two or more flavonoid tinctures, each flavonoid tincture having an amount of a flavonoid and an amount of solvent wherein said flavonoid is dissolved in the solvent, each said flavonoid tincture being majority by weight solvent with an amount of flavonoid in solution and wherein said flavonoid is 1% to 100% saturated in said solvent; and
16. The tincture of claim 15, wherein said flavonoid is resveratrol, pterostilbene, quercetin, procyanidin, curcumin, or combinations thereof.
17. The tincture of claim 15 wherein the solvent is a majority by weight ethanol, dimethyl sulfoxide (DMSO), 1-butanol [n-butanol], 1-propanol, cyclodextrins, or combinations thereof.
18. The tincture of claim 15, further comprising additional additives, wherein said additional additives are water, spices, vitamins and minerals, color additives, flavor additives, or combinations thereof.
19. The tincture of claim 15 wherein the flavonoid concentration in a solvent of 5%-40% ethanol is between 1 and 800 milligrams per liter of ethanol.
20. The tincture of claim 15 wherein the flavonoid concentration in a solvent containing 5%-40% ethanol is between 1 and 800 milligrams per liter of ethanol.
21. The tincture of claim 15 further comprising an amount of one or more additional flavonoid tinctures having an amount a flavonoid and an amount of solvent wherein said flavonoid is dissolved in the solvent, said additional flavonoid tinctures being majority by weight solvent with an amount of flavonoid in solution and wherein said flavonoid is 1% to 100% saturated in said solvent.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0013]
[0014]
[0015]
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0016] The following detailed description is merely exemplary in nature and is in no way intended to limit the scope of the invention, its application, or uses, which may vary. The invention is described with relation to the non-limiting definitions and terminology included herein. These definitions and terminology are not designed to function as a limitation on the scope or practice of the invention, but are presented for illustrative and descriptive purposes only.
[0017] Various terms used throughout the specification and claims are defined as set forth below as it may be helpful to an understanding of the invention.
[0018] As used herein “mixture” shall mean a physical combination of two or more substances on which the identities are retained and are mixed but are not combined chemically.
[0019] As used herein, “food” or “beverage” may alternatively be a food, beverage, cosmetic or drug product.
[0020] As used herein, “ethanol” shall mean pure ethanol or a solvent containing a majority by weight of ethanol.
[0021] As used herein, “solvent” shall mean ethanol (as defined above), DMSO, or other solvent known in the art that is not harmful to humans in small quantities.
[0022] As used herein “tincture” shall mean an alcohol concentrate along with other materials that is used as an additive for a food or beverage.
[0023] As used herein, “flavonoid” shall mean those plant metabolites thought to provide health benefits through cell signaling pathways and antioxidant effects, including resveratrol, pterostilbene and quercetin.
[0024] As used herein “non-alcoholic beverage” shall mean a beverage having an alcoholic content by volume between 0.0% alcohol by volume (ABV) and 2.8% ABV. In some embodiments of the present invention, a “non-alcoholic beverage” is between 0.0% and 0.5% ABV.
[0025] As used herein “alcoholic beverage” shall mean any beverage with ABV content greater than that amount which is considered to be a non-alcoholic beverage.
[0026] As used herein “resveratrol” shall mean trans-resveratrol, cis-resveratrol, or dihydro-resveratrol. Trans-resveratrol having the structure provided in formula (I):
##STR00001##
[0027] As used herein “quercetin” shall mean quercetin having the structure provided in formula (II):
##STR00002##
[0028] As used herein “pterostilbene” shall mean trans-pterostilbene having the structure provided in formula (III):
##STR00003##
[0029] As used herein “bioavailable” shall mean the amount of a particular dose of a drug product that is available within the blood stream for absorption or metabolization. In terms of resveratrol, bioavailable shall mean free trans-resveratrol and/or its conjugate forms or derivatives thereof available in a mixture or tincture.
[0030] As used herein “flavonoid(s)” shall mean resveratrol, quercetin, pterostilbene or combinations thereof.
[0031] The present invention provides a tincture which includes at least one flavonoid and solvent to be infused with a food, beverage, nutraceutical or cosmetic. It should be appreciated that by creating a flavonoid/solvent solution that a bioavailable dose of the flavonoid may be delivered to a subject orally. Without intending to be bound to a particular theory, it is believed that flavonoids, particularly resveratrol and pterostilbene, become hydrogen bonded upon making a solution with solvents, particularly ethanol or DMSO, in the present invention. The hydrogen bonded flavonoid can be added to any food or beverage and provide similar bioavailability as the tincture itself. It is further believed that the hydrogen bonded flavonoid is more easier metabolized by the body, thus causing the flavonoid to be absorbed into the bloodstream and increasing its bioavailability. In at least one embodiment a tincture is made of a solution of a flavonoid, or a combination of flavonoids, immixed with a solvent where the tincture is a homogeneous (single-phase) solution with the flavonoid solute hydrogen bonded to the solvent.
[0032] Several methods for making a tincture to provide a bioavailable amount of flavonoid are provided herein. A tincture of flavonoid and solvent is formed by mixing an amount of flavonoid with an amount of solvent to form a mixture and agitating the flavonoid and solvent mixture dissolving the flavonoid into solution. In at least one embodiment of the present invention, the solvent used is of food grade or pharmacy grade ethanol or DMSO. In at least one embodiment of the present invention, the ethanol or DMSO added to the tincture should be between 5% and 100% ethanol or DMSO. In at least one embodiment of the present invention, the ethanol is at least 160 proof. In at least one embodiment of the present invention, the ethanol is 200 proof.
[0033] It is to be understood that in instances where a range of values are provided that the range is intended to encompass not only the end point values of the range but also intermediate values of the range as explicitly being included within the range and varying by the last significant figure of the range. By way of example, a recited range of from 1 to 4 is intended to include 1-2, 1-3, 2-4, 3-4, and 1-4.
[0034] It should be appreciated that while one may ingest an undiluted bioavailable tincture of flavonoid and ethanol directly, that such a high level of alcohol delivered directly to the body may have harmful effects which counteract the benefits from delivering bioavailable flavonoid. It is understood in the art that pure alcohol involves benzene azeotrope and is generally not for consumption.
[0035] In at least one embodiment of the present invention, the tincture may be added to a food or beverage. In at least one embodiment of the present invention, the tincture is added after all processing, distilling or heating has been performed on a food or beverage. In at least one embodiment of the present invention, the tincture is added to a food or beverage at the point of service of the food or beverage. In at least one embodiment, the tincture in encapsulated in a gel cap or immixed with a pharmaceutically acceptable carrier or gel and delivered as a pill or in a gel cap.
[0036] In some embodiments of the present invention, the tincture may be diluted with water. For example a resveratrol concentration in a solvent of: a) 100% ethanol may be between 0 and 42 grams per liter of ethanol; b) 90% ethanol and 10% water may be between 0 and 32 grams per liter of ethanol; c) 80% ethanol and 20% water may be between 0 and 22 grams per liter of ethanol; d) 70% ethanol and 30% water is between 0 and 13 grams per liter of ethanol; e) 60% ethanol and 40% water is between 0 and 6.5 grams per liter of ethanol; 50% ethanol and 50% water is between 0 and 2.5 grams per liter of ethanol; f) 40% ethanol and 60% water is between 0 and 800 milligrams per liter of ethanol; g) 30% ethanol and 70% water is between 0 and 250 milligrams per liter of ethanol; h) 20% ethanol and 80% water is between 0 and 70 milligrams per liter of ethanol; i) 10% ethanol and 90% water is between 0 and 35 milligrams per liter of ethanol; and j) 5% ethanol and 95% water is between 0 and 18 milligrams per liter of ethanol.
[0037] Other additives may be introduced to the tincture to provide flavoring or other health benefits. In at least one embodiment of the present invention, the tincture may include one or more additional additives. In at least one embodiment of the present invention, additional additives may be water, spices, vitamins and minerals, color additives, flavor additives, or combinations thereof. In at least one embodiment spices may include pumpkin spice, cinnamon, sugar, sage, vanilla bean extract, or combinations thereof. In at least one embodiment, vitamins and minerals may include Thiamine hydrochloride, riboflavin (Vitamin B2), niacin, niacin amide, folate or folic acid, beta carotene, potassium iodide, iron or ferrous sulfate, alpha tocopherols, ascorbic acid, Vitamin D, amino acids (L-tryptophan, L-lysine, L-leucine, L-methionine), or combinations thereof. In at least one embodiment, color additives may include Orange B, Citrus Red No. 2, annatto extract, beta-carotene, grape skin extract, cochineal extract or carmine, paprika oleoresin, caramel color, fruit and vegetable juices or saffron. In at least one embodiment, flavor additives may be vinegar, citric acid, tartaric acid, malic acid, fumaric acid, lactic acid, high fructose corn syrup, hydrolyzed proteins, artificial sweeteners, or food extracts.
[0038] In some embodiments of the present invention the amount of flavonoid delivered in a tincture is to be of sufficient doses that are known to provide the health benefits associated with the selected flavonoid. In at least one embodiment, the flavonoid concentration is between 0.001 and 42 grams per liter of solvent. In at least one embodiment of the present invention, the flavonoid concentration is between 1 and 18 milligrams per liter of solvent. In at least one embodiment of the present invention, 1 to 100 milliliters of a 0.001 to 42 gram flavonoid per liter of solvent tincture is added to a 12 ounce beverage. In at least one embodiment 1 to 10 milliliters of the flavonoid tincture is added to a 12 ounce beverage.
[0039] Not being bound to any particular theory, a 25 mg dose of bioavailable trans-resveratrol is equivalent to 10-25 servings of red wine, understanding that red wines have variable amounts of bioavailable trans-resveratrol. Example 9, for instance, the red wine used for testing bioavailable trans-resveratrol was below the detection limits of the measuring device, while adding the inventive tincture provided 450 times more bioavailable resveratrol than what appeared in the red wine without the tincture.
[0040] It should be appreciated that adding an amount of tincture to a food or beverage may affect the alcohol content for which a food or beverage has been approved, or is desired. Thus, the present invention provides for a method of infusing a tincture of ethanol and flavonoid in a food or beverage, with minimal effect on the alcohol content of the food or beverage. Alternative solvents, such as DMSO may not have the same effects on alcohol content. In at least one embodiment of the present invention, a tincture of solvent and flavonoid is prepared and added to a beverage, and allowed sufficient time for the tincture to infuse with the beverage. In at least one embodiment of the present invention, the amount of tincture added to the beverage changes the amount of alcohol by volume (ABV) of the beverage between 0.01% and 1.0%. In at least one embodiment of the present invention, the amount of tincture changes the ABV of the beverage between 0.02% and 0.5%.
[0041] In certain embodiments of the present invention, the beverage is a non-alcoholic beverage. In at least one embodiment of the present invention, an amount of 10 milligram per liter tincture is added to a 12 ounce non-alcoholic beverage thus providing a bioavailable amount of flavonoid while minimally changing the alcohol by volume of the non-alcoholic beverage from 0.0% to 0.02%. In at least one embodiment of the present invention, a 0.1 milliliter dose of tincture is added to a 12 ounce non-alcoholic beverage thus providing a bioavailable amount of flavonoid while minimally changing the alcohol by volume of the non-alcoholic beverage from 0.0% to 0.02%. In at least one embodiment of the present invention, an amount of 100 milligram per liter tincture is added to a 12 ounce non-alcoholic beverage thus providing a bioavailable amount of flavonoid while minimally changing the alcohol by volume of the non-alcoholic beverage from 0.0% to 0.2%. In at least one embodiment of the present invention, a 1 milliliter dose of tincture is added to a 12 ounce non-alcoholic beverage thus providing a bioavailable amount of flavonoid while minimally changing the alcohol by volume of the non-alcoholic beverage from 0.0% to 0.2%.
[0042] In other embodiments of the present invention, the beverage is an alcoholic beverage. In at least one embodiment an amount of a 10 milligram per liter tincture is added to a 12 ounce alcoholic beverage thus providing a bioavailable amount of flavonoid while minimally changing the alcohol by volume of the alcoholic beverage from 5.0% to 5.02%. In at least one embodiment a 0.1 milliliter dose of tincture is added to a 12 ounce alcoholic beverage thus providing a bioavailable amount of flavonoid while minimally changing the alcohol by volume of the alcoholic beverage from 5.0% to 5.02%. In at least one embodiment an amount of a 100 milligram per liter tincture is added to a 12 ounce alcoholic beverage thus providing a bioavailable amount of flavonoid while minimally changing the alcohol by volume of the alcoholic beverage from 5.0% to 5.2%. In at least one embodiment a 1.0 milliliter dose of tincture is added to a 12 ounce alcoholic beverage thus providing a bioavailable amount of flavonoid while minimally changing the alcohol by volume of the alcoholic beverage from 5.0% to 5.2%.
[0043] In at least one embodiment, the tincture for a beverage infusion with 42 grams of flavonoid and a solvent containing 0% water and 100% solvent with a saturation of 42 g/L. In at least one embodiment a solution containing 42 grams of flavonoid in liter of solvent is passable through a 10 micron filter. In at least one embodiment the amount of bioavailable flavonoid present in a beverage containing the flavonoid tincture is 25 milligrams. In at least one embodiment 0.6 milliliters of tincture is added to a 355 milliliter (12 ounce) beverage to deliver a flavonoid dose of 25 milligrams. In at least one embodiment, the tincture for a food infusion is a solvent of 70% water and 30% ethanol, yielding 250 mg of flavonoid per liter of tincture.
Batch-Mixed Tinctures
[0044] It is appreciated that it is desirable to have one or more flavonoids included in a tincture to provide for a synergistic result from a mixture of flavonoids. It should be understood that by mixing two or more flavonoids first, then exposing the flavonoid mixture to a solvent will result in an imbalance of the flavonoids in solution. Moreover, since solutions in the food and pharmaceutical industry are exposed to a filtration process, many of the intended flavonoids may be filtered out. Without being bound to any particular theory, it is believed that if a flavonoid is not appropriately dissolved or hydrogen bonded to the solvent, then it will remain undissolved, and thus likely will not become bioavailable. This is because certain flavonoids have a higher affinity for dissolving in a solvent, or certain solvents may be stronger solvents than others. As a result, simply mixing flavonoids then mixing with a solvent will likely not result in the desired flavonoid concentration intended.
[0045] Accordingly, a method of mixing an amount of one or more individual tinctures of flavonoids for making batch mixtures of specifically desired combinations of bioavailable flavonoids is provided. The batch-mixture is intended for infusing a beverage, food, cosmetic or drug to provide a bioavailable dose of said flavonoids. Alternatively, the tincture may be administered to a patient or animal directly by direct infusion into a body through known administration methods in the art, including without limit, intravenously, subcutaneously, intradermally, intramuscularly, sublingually, or any other method of parenterally delivered products and including products delivered through various feeding tubes. The method includes creating a first flavonoid tincture by mixing an amount of a flavonoid with an amount of solvent and agitating the flavonoid and solvent mixture until said flavonoid is dissolved in the solvent, then creating a second flavonoid tincture by mixing an amount of a flavonoid with an amount of solvent and agitating the flavonoid and solvent mixture until said flavonoid is dissolved in the solvent. Without being bound to any particular theory, the pre-mixing of the individual tinctures is believed to bond the desired flavonoids to the solvent. The method further includes mixing an amount of the first flavonoid tincture with the second flavonoid tincture to form a batch-mixed tincture solution. The batch-mixed tincture results in the desired concentration of the bioavailable flavonoids despite their differing affinities since the flavonoids are dissolved and believed to be hydrogen bonded to the solvent, thus preventing a preferential dissolution of the flavonoids for given solvents.
[0046] While the foregoing refers to a first flavonoid tincture and a second flavonoid tincture, it is appreciated that multiple flavonoids or multiple solvents may be used to make a batch-mixed tincture, thus nothing herein is intended to limit the batch-mixed tincture to only a first flavonoid tincture and second flavonoid tincture. Illustratively, where five flavonoids are desired to be mixed and only one solvent is to be used, at least five flavonoid tinctures may be made and included in the mixing process to create a batch-mixed tincture. The same is said for tinctures using differing solvents or flavonoids.
[0047] In at least one embodiment the first flavonoid tincture has a different flavonoid than the second flavonoid tincture. In at least one embodiment, the first flavonoid tincture has a different solvent than the second flavonoid tincture. In at least one embodiment, the amount of the first flavonoid tincture and the amount of the second flavonoid tincture to be mixed is determined by calculating the relative concentrations of the flavonoid in the tinctures and mixing the appropriate volumes of the respective tinctures to achieve the desired flavonoid concentrations in the batch-mixed tinctures. In at least one embodiment, the amount of the first flavonoid tincture and the amount of the second flavonoid tincture to be mixed is determined by calculating the relative alcohol by volume of the respective tinctures and mixing the appropriate volumes of the respective tinctures to achieve the desired alcohol by volume of the batch-mixed tincture. It should be appreciated that where different solvents are used, that in some embodiments the flavonoid used in the first and second flavonoid tincture may be the same flavonoid. Similarly, it should be appreciated that where different flavonoids are used, that in some embodiments the solvent used in the first and second flavonoid tincture may be the same solvent.
[0048] Appropriate solvents used for making any tincture shall be pharmaceutical or food grade, but otherwise may be any solvent known in the art. In at least one embodiment, the solvent may include, without limit, ethanol, dimethyl sulfoxide (DMSO), 1-butanol [n-butanol], 1-propanol, cyclodextrins, or combinations thereof. Likewise, flavonoids used for making any tincture may be any flavonoid known in the art. In at least one embodiment, the flavonoid may include, without limit, resveratrol, pterostilbene, quercetin, procyanidin, curcumin, or combinations thereof.
[0049] It is appreciated that in certain embodiments where the first and second tincture are mixed, that the tinctures may not have been pre-filtered prior to making the batch-mixed tincture. Alternatively, it is understood that some amount of undissolved flavonoid, or some amount of precipitated flavonoid may separate from solution after the mixture of the first and second flavonoid tinctures. Thus in at least one embodiment, the batch-mixed tincture solution is filtered after mixing in order to remove any undissolved flavonoid, any impurities, any other suspended solids, or combinations thereof.
[0050] It is appreciated that the intent of the present invention is to provide a bioavailable dose of flavonoids to beverages, foods, cosmetics, or pharmaceuticals. Thus in at least one embodiment, the flavonoid concentration of any flavonoid tincture is between 0.001 and 50 grams per liter of solvent.
[0051] The present invention further includes method for infusing the batch-mixed tinctures as previously described herein. Thus in at least one embodiment, the batch-mixed tincture is added a food, beverage, cosmetic or drug product. It is appreciated that in some instances immediate application of the tincture to a food or beverage may provide a bioavailable dose, but in some embodiments it may be preferred to allow sufficient time for the tincture to infuse with the food, beverage, cosmetic or drug product. This wait-time could be for a homogeneous mixture (or as near as one could be achieved) of the tincture within the food or cosmetic, or providing enough of a sufficient time for an amount of the tincture to distribute within the food or beverage to avoid the addition of the tincture from altering the taste of the food or beverage.
[0052] The present invention further provides a batch-mixed flavonoid tincture. In at least one embodiment the batch-mixed tincture is created using the methods described herein. In at least one embodiment of the batch-mixed tincture, the flavonoid concentration in a solvent of 5%-40% ethanol is between 1 and 800 milligrams per liter of ethanol. In at least one embodiment of the batch-mixed tincture, the flavonoid concentration in a solvent containing 5%-40% ethanol is between 1 and 800 milligrams per liter of ethanol.
[0053] It is further appreciated that the method or the tincture described herein may contain additional additives which are not flavonoids. In at least one embodiment, the tincture includes additional additives of water, spices, vitamins and minerals, color additives, flavor additives, pro-drugs, pharmaceuticals, nutraceuticals, or combinations thereof. In at least one embodiment, the method for making a batch-mixed tincture includes the step of adding to the individual tincture solutions additional additives of water, spices, vitamins and minerals, color additives, flavor additives, pro-drugs, pharmaceuticals, nutraceuticals, or combinations thereof.
EXAMPLES
[0054] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.
Example 1
[0055] A tincture of resveratrol and ethanol is made by immixing resveratrol and ethanol to provide 100 milligrams of resveratrol per liter of ethanol. A beer containing 5.0% ABV is poured into a glass. A direct measure of 0.1 mL dose of tincture is added to the 12 ounce beer. The added tincture raises the ABV of the beer to 5.02% while providing a bioavailable dose of resveratrol.
Example 2
[0056] Example 1 is repeated, but instead using a high-end mega-dose of 1.0 mL of tincture raising the ABV of the beer with tincture to 5.2%.
Example 3
[0057] A tincture of resveratrol and ethanol is made by immixing resveratrol and ethanol to provide 100 milligrams of resveratrol per liter of ethanol. A 12 ounce glass of water is poured and a syringe is used to measure a 0.1 mL dose of tincture and is added to the 12 ounces of water. The addition of the tincture raises the ABV of the water to 0.02% while providing a bioavailable dose of resveratrol.
Example 4
[0058] Example 3 is repeated, but instead using a high-end mega-dose of 1.0 mL of tincture raising the ABV of the water with tincture to 0.2%.
Example 5
[0059]
Example 6
[0060] Example 5 is repeated, but includes the additional step 204 of passing the solution through a 10 micron filter to remove any undissolved resveratrol and any other impurities contained in the resveratrol powder.
Example 7
[0061] A tincture is made containing concentrated (near-saturated at 25° C.) resveratrol and ethanol tinctures of 420 mg/10 mL (42.0 mg/mL). To deliver a 25 mg dose, 0.60 mL of tincture is required regardless of volume of beverage.
Example 8
[0062] A tincture of trans-resveratrol with ethanol is prepared with food grade or pharmaceutical grade 180 to 200 proof ethanol. Observations under multiple environmental settings confirmed the stability of the tincture created. Using medical grade measuring devices, the tincture was added to various beverages in volumes necessary to replicate and exceed the amount of trans-resveratrol in a typical serving of red wine from the Mediterranean stock. Observations demonstrated that there was no precipitation, no change in color, no change in taste and calculations confirmed that the amount of increase in alcohol by volume was maintained below any regulatory standards for required reporting.
Example 9
[0063] A tincture of trans-resveratrol with ethanol is prepared with food grade 200 proof ethanol. The trans-resveratrol tincture is was prepared by adding 421.5 mg trans-resveratrol to 10 mL of 200 proof (100%) food grade ethanol. This resulted in a clear, golden-colored solution with concentration of 42.15 mg/mL.
[0064] A dose of 25 mg of trans-resveratrol is delivered to a serving volume of various drinks including red wine, white wine, beer (Yuengling), spirits such as vodka and bourbon (Jim Beam), energy drinks (5 Hour Energy), sodas (diet Coke, Coke, Sprite) and orange juice (Tropicana). To deliver 25 mg of trans-resveratrol, 0.60 milliliters of trans-resveratrol tincture is required (42.15 mg/mL×0.60 mL=25.29 mg). An amount of the trans-resveratrol tincture is added to each beverage and is calculated based on the volume of the serving. The serving volume for a glass of wine is 187 mL. The serving volume for a glass of wine plus the 0.60 mL of trans-resveratrol tincture was divided by the dose of trans resveratrol 25.29 mg/187.6 mL=0.1348 mg/mL. 0.1348 mg/mL was then multiplied by 1000 ug/mg to get the amount in ug/mL, 0.1348 mg/mL×1000 ug/mg=134.8 ug/mL. Similar calculations are performed for the remaining beverages. For beer, sodas and orange juice, the serving volume is 355 mL for each beverage, resulting in 71.1 ug/mL of trans-resveratrol to be infused into each beverage. For spirits, the serving volume for spirits is 50 mL, resulting in 499.8 ug/mL of trans-resveratrol infused. For Five Hour Energy®, the serving volume is 57 mL, resulting in 439.1 ug/mL of trans-resveratrol infused. A red wine blank was used as a control in order to show natural resveratrol being present in the beverage.
[0065] After the calculations are completed a syringe is used to transfer 0.6 mL of trans-resveratrol tincture to a serving volume of each beverage. The tincture is delivered to each serving volume below the surface of the liquid. In all cases the trans-resveratrol tincture appears to dissolve as a homogenous solution. All beverages receiving the tincture are thoroughly mixed, and a few milliliters of each are transferred to amber-colored sample vials. Samples are stored from each beverage in a refrigerator for analysis.
[0066] During analysis the percent of the free trans-resveratrol recovered in the various drinks was determined via HPLC/(+)ESI-MS/MS and HPLC/UV (305 nm).
[0067] The results of this experiment are presented in Table 1.
TABLE-US-00001 TABLE 1 [t-Rsv] (ug/mL) % Theory Drink# Sample: Theor MS/MS UV MS/MS UV 1 red wine blank 0 <<<spike n/a n/a 2 Red Wine, spiked 134.8 45.8 100.0 33.9 74.2 3 White wine spiked 134.8 63.8 108.5 47.4 80.5 4 Beer; Yuengling 71.1 34.0 74.3 47.8 104.5 Summer Wheat 5 Vodka (V), AB 499.8 489.3 549.0 97.9 109.9 Brand 6 Bourbon (BO), 499.8 584.0 534.6 116.9 107.0 Jim Beam 7 5H, Grape 439.1 470.4 473.8 107.1 107.9 8 Diet Coke 71.1 57.1 68.7 80.2 96.6 9 Coke 71.1 49.1 64.7 69.0 90.9 10 Sprite 71.1 53.8 61.4 75.7 86.3 11 OJ, Tropicana 71.1 21.1 41.4 29.7 58.2
[0068] It is observed that all beverages receiving the tincture show measurable concentrations of trans-resveratrol. The red wine blank, which is not spiked, is analyzed for natural trans-resveratrol background. While trans-resveratrol is detected in the red wine blank, it is at the instrument's minimum detection limit. Integration of peak areas show the ratio of spiked red wine to unspiked red wine was ˜450. Moreover, the trans-resveratrol in solution is bioavailable because the detection methods can only detect free and unbound trans-resveratrol.
[0069] Generally, UV detection reveals higher trans-resveratrol concentrations than did MS/MS detection. MS/MS detection focuses on the 228 mass number, resveratrol's molecular weight. UV detection at a fixed wavelength (305 nm) shows detection of the molecule's conjugated core which includes the two aromatic rings and the adjoining C═C pi bond. Notwithstanding, the recovery values were generally moderate 30-70% to high 70-100%.
Example 10
[0070] A tincture of trans-resveratrol with ethanol is prepared with food grade 200 proof ethanol. To deliver 25 mg of trans-resveratrol, 0.60 milliliters of trans-resveratrol tincture is required (42.15 m/mL×0.60 mL=25.29 mg). The serving volume for spirits is 50 mL. 25.29 mg/50.6 mL resulting in 499.8 ug/mL of trans-resveratrol to infuse the spirits with. A syringe is used to transfer 0.6 mL of trans-resveratrol tincture into the vodka and bourbon. The infused drinks show measurable concentrations of trans-resveratrol. The percent recovery of free trans-resveratrol recovered in the vodka during analysis is 97.9% MS/MS and 109.9% UV, and in the bourbon is 116.9% MS/MS and 107% UV. It is appreciated that the percent recovery in some instances is greater than 100% because of +/−17% error because of the techniques used to measure trans resveratrol. As a result these values can be considered to be 100% recovery in the vodka and bourbon beverages. These results show the bioavailability of the trans-resveratrol tincture added to beverages.
Example 11
[0071] 25 mg of trans-resveratrol powder is added to a serving volume of various drinks including red wine, white wine, beer (Yuengling), spirits such as vodka and bourbon (Jim Beam), energy drinks (5 Hour Energy), sodas (diet Coke, Coke, Sprite) and orange juice (Tropicana). In all cases the trans-resveratrol appears to suspend in solution forming a cloudy layer. All beverages receiving the powder are thoroughly mixed. Each beverage is passed through a 10 micron filter to remove any undissolved resveratrol and any other impurities contained in the resveratrol powder. A few milliliters of each beverage are transferred to amber-colored sample vials. Samples are stored from each beverage in a refrigerator for analysis.
[0072] During analysis the percent of the free trans-resveratrol recovered in the various drinks was determined via HPLC/(+)ESI-MS/MS and HPLC/UV (305 nm). It is observed that none of the non-alcoholic beverages (energy drink, sodas, and orange juice) receiving the powder show any measurable concentrations of trans-resveratrol. This differs from the results provided in Example 9 drastically, showing that merely adding powder resveratrol to a beverage does not provide a bioavailable amount of resveratrol. Notwithstanding, it was observed that the alcoholic beverages (red wine, white wine, beer and spirits) receiving the powdered resveratrol did have some amount of bioavailable resveratrol, but below the minimum detection limits of the devices, and thus could not be quantified. This again shows a drastic difference from the results of Example 9, confirming that merely adding resveratrol powder to a beverage minimally delivers a bioavailable amount of resveratrol as compared to the inventive tincture.
Example 12
[0073] Examples 1 through 9 are repeated using the solvent DMSO and the flavonoid pterostilbene.
Example 13
[0074] A tincture of pterostilbene and solvent is made by dissolving approximately 420 mg trans-pterostilbene powder into 10 mL of pharmacy grade ethanol.
Example 14
[0075] A tincture of resveratrol, pterostilbene, and ethanol is made by combining separate volumes of tincture of resveratrol and tincture of pterostilbene in a range of the following volume resveratrol to volume pterostilbene ratios: 9:1, 4:1, 7:3, 3:2, 1:1, 2:3, 3:7, 1:4, and 1:9.
[0076] A similar study of that of Example 9 was conducted using above combinations of resveratrol and pterostilbene. The results of this experiment are presented in Table 2.
TABLE-US-00002 TABLE 2 Binary Tincture Range of Formulation: R + Pt (vol R:vol Pt) Conc (mg/mL) % R % Pt Ratio R:Pt Conc (mg/mL) R Pt 100 0 Pure R 42.0 0.0 90 10 9:1 37.8 4.2 80 20 4:1 33.6 8.4 70 30 7:3 29.4 12.6 60 40 3:2 25.2 16.8 50 50 1:1 21.0 21.0 40 60 2:3 16.8 25.2 30 70 3:7 12.6 29.4 20 80 1:4 8.4 33.6 10 90 1:9 4.2 37.8 0 100 Pure Pt 0.0 42.0
Example 14
[0077] A 1.0 mL volume of the resulting tincture in Example 13 contains the following range of mg doses of resveratrol and pterostilbene respectively: 37.8, 4.2; 33.6, 8.4; 29.4, 12.6; 25.2, 16.8; 21.0, 21.0; 16.8, 25.2; 12.6, 29.4; 8.4, 33.6; and 4.2, 37.8.
Example 15
[0078] A tincture of resveratrol, quercetin, and ethanol is made by dissolving 1.5 mg of quercetin powder into 1.0 mL of tincture of resveratrol. The resulting tincture contains 42 mg resveratrol and 1.5 mg quercetin.
Example 16
[0079] A tincture of resveratrol, pterostilbene, and quercetin is made by dissolving 1.5 mg of quercetin powder into 1.0 mL of the tincture of resveratrol, pterostilbene, and ethanol of any ratio over the range cited in Example 13.
Example 17
[0080] A 100% saturated ethanol based tincture of R mixed 1:1 (equal volumes) with a 100% saturated ethanol based tincture of Curcumin yields a batch-mixed tincture with 50 mg of Resveratrol and 10 mg of Curcumin per volume of tincture. The batch-mixed tincture would yield a concentration of 25 mg/mL Resveratrol and 5 mg/mL Curcumin.
Example 18
[0081] A 100% saturated ethanol based tincture of Resveratrol mixed 1:1 (equal volumes) with a 100% saturated DMSO based tincture of Curcumin yields a batch-mixed tincture with 50 mg of Resveratrol and 25 mg of Curcumin per volume of tincture and a ratio of ethanol to DMSO of 1:1 by volume. The batch-mixed tincture would yield a concentration of 25 mg/mL Resveratrol and 12.5 mg/mL Curcumin in the co-solvent (ethanol/DMSO) solution.
Example 19
[0082] Combinations of Examples 17 and 18 will yield various ratios of resveratrol, curcumin, ethanol and DMSO, one combination being a tincture that yields a ratio of ethanol to DMSO of 3:1 by volume, and a ratio of resveratrol to curcumin of 100:35 by mass
Other Embodiments
[0083] While at least one exemplary embodiment has been presented in the foregoing detailed description, it should be appreciated that a vast number of variations exist. It should also be appreciated that the exemplary embodiment or exemplary embodiments are only examples, and are not intended to limit the scope, applicability, or configuration of the described embodiments in any way. Rather, the foregoing detailed description will provide those skilled in the art with a convenient road map for implementing the exemplary embodiment or exemplary embodiments. It should be understood that various changes can be made in the function and arrangement of elements without departing from the scope as set forth in the appended claims and the legal equivalents thereof.