Methods of oxidizing HOPS and products therefrom

Abstract

Described is a method to generate oxidized hops compounds in an aqueous solution using ozone. The result of this method is a composition of oxidized hops compounds which can be used to induce thermogenesis in humans by activating brown adipose tissue, a type of fat found in the human body, which would allow calories to be burned while drinking alcoholic beverages.

Description

[0001] METHODS OF OXIDIZING HOPS AND PRODUCTS THEREFROM

[0002] CROSS REFERNCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 468,405, filed on May 23, 2023, which is incorporated by reference herein in its entirety.

[0004] BACKGROUND

[0005] Hops are the flower of the hop plant Humulus lupus), which has been used as an ingredient in brewing beer for well over a decade. The Association of Brewing Chemists has well established criteria for the evaluation and use of hops in the flavoring of beers. Hops impart preservative properties to the beer through antimicrobial activity against bacteria and fungus, and are responsible for the retention of the beer’ s foaming head. The chemical compounds within hops contribute to the flavor and aroma profile of beers, adding bitter notes, which are desirable characteristics of beers.

[0006] Among the compounds produced within the lupulin glands of hops, which are in the flowering cones, are volatile hydrocarbons known as essential oils and resinous amphiphilic compounds known as a- and P-acids, which give beer a bitter taste. This bitter taste is obtained by boiling the acids in a slightly acidic wort, a mixture of water and unfermented maltose, forming primarily a mixture of isomerized compounds. Initially, the a-acids isomerize to their iso a-acid derivatives, which continue to slowly oxidize. The P-acids are insoluble in boiling wort and as a result, take a longer time to oxidize.

[0007] Both clinical and empirical studies have shown that isolated oxidized hops a- and P- acids, when concentrated, can promote weight loss, among other benefits. Beer contains a small percentage, if any, of oxidized a- and P-acids and thus does not promote weight loss. Quite the opposite, science has shown that after drinking alcohol, the alcohol is metabolized first to provide energy while other calories that are consumed are stored as fat. Until the alcohol is completely metabolized, any calories consumed will be stored as fat. This process continues for many hours after drinking. Alcohol, even when consumed in moderation, can stay in a person’s system for up to 24 hours thus making it easy to accumulate fat.

[0008] Although the alcoholic beverage industry has gone to great lengths to lessen the effects of a slow metabolism by reducing carbohydrates, calories and sugars within certain alcoholic beverages, alcohol itself is primarily responsible for increases in belly fat. Thus, the only significant way to prevent alcohol’s impact on metabolism is to avoid alcohol altogether. What is thus needed are new compositions and methods that can reduce the effect of alcohol on gaining body fat. The compositions and methods disclosed herein can address these and other needs.

[0009] SUMMARY

[0010] Disclosed herein is a method to generate oxidized hops compounds in an aqueous solution using ozone. The result of this method is a composition of oxidized hops compounds which can induce thermogenesis in humans by activating brown adipose tissue, a type of fat found in the human body, which would allow calories to be burned while drinking alcoholic beverages. The effect of these concentrated oxidized hops compounds on the activation of brown adipose tissue is immediate and long lasting.

[0011] Additional advantages will be set forth in part in the description that follows, and in part will be obvious from the description, or may be learned by practice of the aspects described below. The advantages described below will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive.

[0012] BRIEF DESCRIPTION OF THE FIGURES

[0013] The accompanying figures, which are incorporated in and constitute a part of this specification, illustrate several aspects described below.

[0014] Figure 1 is an example chromatogram of a standard injection. The elution order is Cohumulone (10.94), Coeluting Adhumulone and Humulone (14.14), Colupulone (19.51) and Coeluting Adlupulone and Lupulone (25.81).

[0015] Figure 2 is an example chromatogram of the hops starting material prior to the oxidation process.

[0016] Figure 3 is an example overlay chromatogram of the hops starting material and the standard.

[0017] Figure 4 is an example chromatogram of the reaction mixture within 30 minutes of starting the oxidation process. Figure 5 is an example chromatogram of the reaction mixture within 60 minutes of starting the oxidation process.

[0018] Figure 6 is an example chromatogram of the reaction mixture within 5 hours of starting the oxidation process.

[0019] DETAILED DESCRIPTION

[0020] The compositions and methods described herein may be understood more readily by reference to the following detailed description of specific aspects of the disclosed subject matter, and the Examples included therein.

[0021] Before the present compositions and methods are disclosed and described, it is to be understood that the aspects described below are not limited to specific synthetic methods or specific reagents, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting.

[0022] General Definitions

[0023] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of particular embodiments, preferred embodiments of compositions, methods and materials are described herein. For the purposes of the present disclosure, the following terms are defined below. Additional definitions are set forth throughout this disclosure.

[0024] The articles “a,” “an,” and “the” are used herein to refer to one or to more than one (i.e., to at least one, or to one or more) of the grammatical object of the article.

[0025] The use of the alternative (e.g., “or”) should be understood to mean either one, both, or any combination thereof of the alternatives.

[0026] As used herein, the term “about” or “approximately” refers to a quantity, level, value, number, frequency, percentage, dimension, size, amount, weight, or length that varies by acceptable levels in the art. In some embodiments, the amount of variation may be as much as 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% to a reference quantity, level, value, number, frequency, percentage, dimension, size, amount, weight, or length. In one embodiment, the term “about” or “approximately” refers a range of quantity, level, value, number, frequency, percentage, dimension, size, amount, weight, or length ± 15%, ± 10%, ± 9%, ± 8%, ± 7%, ± 6%, ± 5%, ± 4%, ± 3%, ± 2%, or ± 1% about a reference quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length.

[0027] A numerical range, e.g., 1 to 5, about 1 to 5, or about 1 to about 5, refers to each numerical value encompassed by the range. For example, in one non-limiting and merely illustrative embodiment, the range “1 to 5” is equivalent to the expression 1, 2, 3, 4, 5; or 1.0,

[0028] 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or 5.0; or 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4,

[0029] 4.5, 4.6, 4.7, 4.8, 4.9, or 5.0.

[0030] By “alpha acid” or “a-acid” is meant an organic acid derived from a hop plant (Humulus lupulus) having structural homology to a humulone, adhumulone, cohumulone, or an analog or derivative thereof. Humulone, adhumulone, and cohumulone are the three most abundant alpha acids. Other exemplary derivatives of an alpha acid include, but are not limited to, iso-alpha acids, rhoisoalpha acids, tetrahydroisoalpha acids, and hexahydroisoalpha acids. Isohumulones, a type of iso-alpha acid, are present as both the cis and trans stereoisomers. Unless otherwise stated, the term is meant to include both stereoisomers.

[0031] By “beta acid” or “(3-acid” is meant an organic acid derived from a hop plant (Humulus lupulus) having structural homology to a lupulone, adlupulone, colupulone or an analog or derivative thereof. Lupulone, adlupulone, and colupulone are the three most abundant beta acids.

[0032] By “oxidized alpha and beta acids” is meant any oxidized form of alpha, isoalpha, and / or beta acid, and can include any degree or amount of oxidization of any one or more alpha, isoalpha, or beta acid, unless specifically stated. “Oxidized alpha and beta acids” are sometimes also referred to herein as oxidized hops compounds, an oxidized hops product, or mature hops bitter acids.

[0033] By “increase” or other forms of the word, such as “increasing,” is meant promoting an event or characteristic (e.g., fat burning). It is understood that this is typically in relation to some standard or expected value, in other words it is relative, but that it is not always necessary for the standard or relative value to be referred to. For example, “increases fat burning” means increasing the rate or amount of fat burning relative to a standard or a control.

[0034] By “reduce” or other forms of the word, such as “reducing” or “reduction,” is meant lowering of an event or characteristic (e.g., weight gain). It is understood that this is typically in relation to some standard or expected value, in other words it is relative, but that it is not always necessary for the standard or relative value to be referred to. For example, “reduces weight gain” means reducing the rate or amount of weight gain relative to a standard or a control.

[0035] Reference will now be made in detail to specific aspects of the disclosed materials, compounds, compositions, articles, and methods, examples of which are illustrated in the accompanying example.

[0036] Methods of Oxidizing

[0037] Disclosed herein, in one aspect, is a method of providing oxidized alpha and beta acids comprising contacting a hops mixture comprising alpha acids, beta acids, and / or iso-alpha acids with ozone, to thereby provide the oxidized alpha and beta acids.

[0038] Hops

[0039] The hops mixture comprising alpha acids, beta acids, and / or iso-alpha acids can be a hops extract, a mixture of water with hops pellets or hops cones, or a raw mixture of hops, hops pellets, or hops cones. In one exemplary method, the hops mixture is a raw mixture of hops, pellets, or cones. When using a hops extract, hops pellets or cones are contacted with water under conditions to extract the alpha acids, beta acids, and / or iso-alpha acids from the pellets or cones. Conditions to extract the acids typically involve temperatures of from above ambient to about 90 °C, e.g., from about 35 °C to about 85 °C, from about 50 °C to about 60 °C. In some examples, the hops pellets or cones are extracted at about 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90°C, where any of the stated values can form an upper or lower endpoint of a range. Extracting the alpha acids, beta acids, and / or iso-alpha / beta acids from the hops pellets or cones can also involve mixing or agitating with mechanical stirrers, shaking, or sonication. In still further examples, after contacting the hops pellets or cones with water, any insoluble debris can be filtered off.

[0040] Unlike other methods for providing oxidized alpha acids from hops, the disclosed methods do not require any pre-treatment of the raw hops material (i.e., hops cones or pellets) prior to oxidization. Other methods require the raw hops materials to be processed so that the alpha acids, beta acids and / or iso alpha acids are obtained as an extract, which may or may not be converted to a powder prior to oxidization. In such cases, the raw materials are macerated, filtered, and then extracted using organic solvents such as ethanol, methanol, or another organic solvent. Another method is to grind the raw hops cones or pellets and extract the desired compounds using super critical carbon dioxide extraction. The hops extract may then be converted to a powder by combining it with another material such as polysaccharides, cellulose derivatives, or another suitably acceptable carrier through processes such as spray-drying, freeze drying, or other drying methods well known to those skilled in the art.

[0041] In contrast to these methods, the disclosed methods can use a simple hops mixture of raw hops material, whether in the form of the hops cones or as pre-formed pellets, as-is, in the commercially available form, without additional processing, grinding, special extraction, or conversion to a powder prior to the oxidation method.

[0042] Ozone

[0043] Ozone is the triatomic, allotropic form of oxygen having the chemical symbol O3 and a molecular weight of 47.998 g / mol. These three oxygen atoms form a relatively unstable, highly oxidative molecule that serves as a strong oxidant. Ozone produced commercially for oxidation reactions is produced as a gas, from air at concentrations from about 1.5 to about 2.0 % by weight in air, or from oxygen at concentrations greater than about 6% and up to about 12% by weight. As ozone is highly reactive and has a short half-life, it is difficult to store and transport. Consequently, ozone is normally generated onsite for immediate use. Thus, quantities of ozone for use in the disclosed methods can be produced with a commercially available ozone generator. The generator works by passing a high voltage, alternating electric discharge through a gas stream containing oxygen.

[0044] In the disclosed methods, ozone can be bubbled or streamed into or onto the hops mixture comprising alpha acids, beta acids, and / or iso alpha acids. This can be done by connecting an ozone generator’s output into or onto the hops mixture as disclosed herein. Alternatively, an ozone generator can be used to dissolve ozone into water, even up to saturating water with ozone. The resulting ozone-containing water can then be contacted to the hops mixture, either by adding the hops mixture comprising alpha acids, beta acids, and / or iso alpha acids into the ozone-containing water, or vice versa. That is, ozone-containing water can be contacted directly with hops pellets or hops cones, either by adding the pellets or cones to the ozone-containing water or pouring the ozone-containing water onto the hops pellets or cones.

[0045] The resulting mixture of ozone and hops mixture can be mixed, agitated, sonicated, or shaken to ensure mixing. The hops mixture can be contacted with the ozone (including ozonecontaining water) for from a few minutes to a day. The contact time between the hops mixture and ozone can be adjusted based on the batch size. A benefit of the disclosed methods is that oxidation of the alpha acids, beta acids, and / or iso alpha acids in the hops mixture is rapid and typically occurs within a few hours. So, in some examples, ozone (or ozone-containing water) can be contacted with the hops mixture for from about 1 hour to about 12 hours, from about 3 hours to about 8 hours, or from about 4 hours to about 6 hours. In further examples, ozone (or ozone-containing water) is contacted with the hops mixture for about 2, about 4, about 6, about 8, about 10, or about 12 hours, where any of the stated values can form an upper or lower end point of a range. In further examples, ozone (or ozone-containing water) is contacted with the hops mixture for up to about 2 hours. Further, in the disclosed methods it is not required to heat the reaction mixture comprising ozone and hops mixture. The oxidation can thus be performed at ambient temperatures, or even below ambient temperatures such as about 4, about 6, about 8, about 10, about 12, about 14, about 16, about 18, about 20, or about 22 °C, where any of the stated values can form an upper or lower endpoint of a range.

[0046] The extent of the oxidation process between the ozone and hops mixture can be monitored by high pressure liquid chromatography (HPLC). After the appropriate contact time between the hops mixture and ozone has been determined, the remaining hops vegetative material can be separated from the concentrated liquid product via a strainer and a series of filtration and / or centrifugation steps. Centrifuges and strainers have a long history of allowing breweries to perform critical activities such as pre-clarification, hot wort separation, green beer separation and beer recovery. Clarification of the final liquid product through separation from the vegetative material helps increase stability. While centrifugation provides more concentrated solids as well as swift removal of the residual vegetative matter, gravity settling may also be utilized.

[0047] Concentration of the final product can be controlled by adjusting the ratios of hops to purified water during manufacturing. Additionally, the concentration of the oxidized hops compounds may be adjusted by dilution after manufacturing. Such adjustments are well known to those skilled in the art.

[0048] In other methods disclosed herein, the oxidized hops compounds can be then sterilized, pasteurized, or homogenized, though such further processing steps are optional.

[0049] These use of ozone in the disclosed methods yield highly purified and sterile water for the oxidized product which promotes long term stability. The disclosed methods eliminate the need for additional preservatives in the finished product. Further, the disclosed methods are unique in that they do not use any organic solvents, other oxidizing agents (c.g., metals or organometallics), or any applied heat at any stage of the manufacturing process to promote the oxidization of the alpha acids, beta acids, or iso alpha acids. In this sense they are also environmentally friendly as well as nutritionally acceptable.

[0050] Compositions

[0051] The disclosed methods can yield a mixture of oxidized alpha and beta acids from hops (e.g., a hops mixture, hops pellets, hops cones), which can then be used without further modification as a virtually odorless and tasteless additive that can be added in small quantities to an alcoholic beverage. Since the product lacks the distinctive aroma and intense bitter taste of hops, it can also be used as an additive for non-alcoholic beverages such as nutritional drinks, dietary shakes, and even water. The resulting oxidized hops compounds of the disclosed methods can contain a concentrated level of oxidized hops compounds, primarily oxidized alpha and beta acids, that can promote weight loss by activating brown adipose tissue to bum calories through thermogenesis even in the presence of alcohol.

[0052] As the disclosed methods use ozone to oxidize the alpha acids, beta acids, and / or iso alpha acids in a hops mixture, and as the disclosed methods are very effective at the oxidation reactions, the resulting products of the disclosed methods comprise oxidized alpha and beta acids. Thus, the amount of alpha acids, beta acids, and / or iso alpha acids in the reaction product of the disclosed methods is reduced as compared to the amount in the starting hops mixture. Likewise, the amount of oxidized alpha and beta acids in the reaction products of the disclosed methods is increased as compared to the amount in the starting hops mixture. The amounts of oxidized alpha and beta acids in the reaction product can be determined by the HPLC method disclosed in Rostad, S. and Sahlstrom, A. Concordia College Journal of Analytical Chemistry 2 (20110, 78 - 83, which is incorporated by reference herein in its entirety for its teachings of analytical methods.

[0053] The method for quantitating the oxidized hops compounds is as follows. Prior to beginning the oxidation process, the hops starting material can be analyzed to determine the total content of alpha acids, beta acids, and iso alpha acids. The quantitation of these components can be accomplished using standards and the corresponding methods for ICE-4 and DCHA-Iso / ICS-14 which can be obtained from the American Society of Brewing Chemists (ASBC). Once the hops starting material is exposed to ozone, the content of alpha acids, beta acids, and iso alpha acids decreases as the oxidation products increase. As a result, the oxidation products can be evaluated by comparing the peak areas of the alpha acids, beta acids, and iso alpha acids in the hops starting material to the total area of the oxidation product. The total peak area is defined as the area from about 2 minutes to about 50 minutes. The peak area ratio is calculated as follows: (Total peak area I Total peak area of alpha acids + iso alpha acids + beta acids). The peak area ratio of the total peak area of the oxidation product to the alpha acids, beta acids, and iso alpha acids as determined by HPLC is about 1.9.

[0054] In some specific examples, disclosed are compositions comprising greater than about 20 % by weight as determined by HPLC of total oxidized alpha acids. In further examples, disclosed are compositions comprising greater than about 20, 30, 40, 50, 60, 70, 80, 90, or 95 % by weight as determined by HPLC of total oxidized alpha acids, where any of the stated values can form an upper or lower endpoint of a range. In other examples, disclosed are compositions comprising from about 20% to about 95%, from about 30% to about 80%, from about 40% to about 70%, from about 50% to about 60%, from about 20% to about 50%, or from about 30% to about 60% by weight as determined by HPLC of total oxidized alpha acids. In a specific example, disclosed are compositions comprising about 33% by weight as determined by HPLC of total oxidized alpha acids.

[0055] In some specific examples, disclosed are compositions comprising greater than about 20 % by weight as determined by HPLC of total oxidized iso alpha acids. In further examples, disclosed are compositions comprising greater than about 20, 30, 40, 50, 60, 70, 80, 90, or 95 % by weight as determined by HPLC of total oxidized iso alpha acids, where any of the stated values can form an upper or lower endpoint of a range. In other examples, disclosed are compositions comprising from about 20% to about 95%, from about 30% to about 80%, from about 40% to about 70%, from about 50% to about 60%, from about 20% to about 50%, or from about 30% to about 60% by weight as determined by HPLC of total oxidized iso alpha acids. In a specific example, disclosed are compositions comprising about 33% by weight as determined by HPLC of total oxidized iso alpha acids.

[0056] In some specific examples, disclosed are compositions comprising greater than about 20 % by weight as determined by HPLC of total oxidized beta acids. In further examples, disclosed are compositions comprising greater than about 20, 30, 40, 50, 60, 70, 80, 90, or 95 % by weight as determined by HPLC of total oxidized beta acids, where any of the stated values can form an upper or lower endpoint of a range. In other examples, disclosed arc compositions comprising from about 20% to about 95%, from about 30% to about 80%, from about 40% to about 70%, from about 50% to about 60%, from about 20% to about 50%, or from about 30% to about 60% by weight as determined by HPLC of total oxidized beta acids. In a specific example, disclosed are compositions comprising about 33% by weight as determined by HPLC of total oxidized beta acids.

[0057] In some specific examples, disclosed are compositions comprising greater than about 20 % by weight as determined by HPLC of total oxidized hops compounds (i.e., oxidized alpha, oxidized beta, and oxidized iso alpha acids). In further examples, disclosed are compositions comprising greater than about 20, 30, 40, 50, 60, 70, 80, 90, or 95 % by weight as determined by HPLC of total oxidized hops compounds (i.e., oxidized alpha, oxidized beta, and oxidized iso alpha acids), where any of the stated values can form an upper or lower endpoint of a range. In other examples, disclosed are compositions comprising from about 20% to about 95%, from about 30% to about 80%, from about 40% to about 70%, from about 50% to about 60%, from about 20% to about 50%, or from about 30% to about 60% by weight as determined by HPLC of total oxidized hops compounds (i.e., oxidized alpha, oxidized beta, and oxidized iso alpha acids). In a specific example, disclosed are compositions comprising about 33% by weight as determined by HPLC of total oxidized hops compounds (i.e., oxidized alpha, oxidized beta, and oxidized iso alpha acids).

[0058] In some specific examples, disclosed are compositions comprising less than about 20 % by weight as determined by HPLC of total alpha acids, e.g., humulone, cohumulone, and adhumulone. In further examples, disclosed are compositions comprising less than about 20, 10, or 5 % by weight as determined by HPLC of alpha acids, where any of the stated values can form an upper or lower endpoint of a range.

[0059] In some specific examples, disclosed are compositions comprising less than about 20 % by weight as determined by HPLC of total beta acids, e.g., lupulone, colupulone, and adlupulone. In further examples, disclosed are compositions comprising less than about 20, 10, or 5 % by weight as determined by HPLC of beta acids, where any of the stated values can form an upper or lower endpoint of a range. In the disclosed compositions, the balance of the composition can comprise water, e.g., about 80, 70, 60, 50, 40, 30, 20, 10, or 5 % water. For example, in some compositions disclosed herein the composition comprises 66% water.

[0060] In still other examples, the disclosed compositions can be substantially free (i.e., having less than about 10%, 5%, 2%, or 1%) of sugars, dextrin, and maltodextrin.

[0061] Still further, disclosed herein are delivery devices comprising the disclosed composition of oxidized alpha acids that can be used to add the compositions to beverages. For example, containers comprising from about 1 to about 10 mL of a solution comprising an oxidized alpha acid are disclosed. Containers can also contain about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10 mL of oxidized alpha and beta acids, where any of the stated values can form an upper or lower endpoint of a range.

[0062] Also, disclosed herein are supplements comprising the compositions disclosed herein. A nutritional supplement is any composition that can be administered to or taken by a subject to provide, supply, or increase a nutrient(s) (e.g., vitamin, mineral, essential trace element, amino acid, peptide, nucleic acid, oligonucleotide, lipid, cholesterol, steroid, carbohydrate, and the like). In one aspect, disclosed herein are nutritional supplements comprising any of the oxidized hopds compounds disclosed herein. For example, a nutritional supplement can comprise a composition comprising one or more oxidized alpha and beta acids from hops.

[0063] The supplement can also comprise other nutrients such as vitamins, trace elements, minerals, and the like. Further, the supplement can comprise other components such as preservatives, antimicrobials, anti-oxidants, chelating agents, thickeners, flavorings, diluents, emulsifiers, dispersing aids, and / or binders.

[0064] Methods of use

[0065] There are two types of fat in the human body, white adipose tissue (WAT) and brown adipose tissue (BAT). White adipose tissue is composed primarily of fatty acids, triglycerides, and cholesterol. WAT stores energy in the form of fat which accumulates in the body causing weight gain.

[0066] Unlike white adipose tissue, BAT contains lots of mitochondria which are responsible for generating heat. During a process called thermogenesis, brown adipose tissue can burn a lot of calories as it produces heat to warm the body, a response especially helpful to cold environments. Brown adipose tissue promotes weight loss by burning calories as it transfers energy from food and converts it into heat during thcrmogcncsis.

[0067] In addition to its role in thermo-regulation, BAT also aids in glucose and lipid metabolism which can help regulate blood sugar levels thereby curbing hunger and cravings. BAT is also involved in the production of leptin, a hormone that affects appetite control.

[0068] Isomerized and oxidized hops compounds have been directly linked to the activation of BAT, and clinically proven to significantly contribute to the fat burning process when consumed daily over time. Morimoto-Kobayashi Y, et al., PLOS One 2015, 10(6): e0131042. doi:10.1371 / journal.pone.0131042.

[0069] Thus, in one aspect, disclosed herein is a process using the oxidized alpha and beta acids to reduce weight gain and / or reduce weight by administering the disclosed oxidized alpha and beta acids to a subject (e.g., human). The oxidized alpha and beta acids can be given with food or a beverage, especially with an alcoholic beverage, either as a separate composition or by adding the oxidized alpha and beta acids into the food or beverage. The oxidized alpha and beta acids can also be administered to a subject neat. The disclosed oxidation method provides an oxidized hops product that provides an immediate and targeted approach to the activation of brown adipose tissue via the digestive system. As a result, calories can be burned in the presence of alcohol instead of being stored as fat. Thus, promoting weight loss.

[0070] EXAMPLES

[0071] The following example is put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how the compounds, compositions, and / or methods claimed herein are made and evaluated, and are intended to be purely exemplary of the invention and are not intended to limit the scope of what the inventors regard as their invention. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.

[0072] Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in °C or is at ambient temperature, and pressure is at or near atmospheric. A calculated quantity of hops pellets were added to the strainer assembly within the tank for about a 10% weight to volume ratio. Filtered purified water was passed through the ozone generator and then through the strainer assembly. The resulting liquid was recirculated through the system for two hours. During the oxidation process, samples were taken over time and analyzed by HPLC to determine the extent of the reaction (See Figs. 2-6). Once the reaction was deemed complete, the ozone generation was discontinued, and the strainer assembly was allowed to drain. All liquid fractions were allowed to settle either by gravity or centrifugation. After the settling period, the oxidation product was further clarified by a series of filtration steps prior to packaging.

[0073] Specific embodiments

[0074] In some aspects, the techniques described herein relate to a method of providing oxidized alpha and beta acids, including: contacting a hops mixture including alpha acids, beta acids, and / or iso-alpha acids with ozone, to thereby provide oxidized alpha and beta acids.

[0075] In some aspects, the techniques described herein relate to a method, wherein the hops mixture is provided by contacting hops pellets with water under conditions to extract the alpha acids, beta acids, and / or iso-alpha acids from the hops pellets.

[0076] In some aspects, the techniques described herein relate to the method of any one of the previous claims, further including filtering the water.

[0077] In some aspects, the techniques described herein relate to the method of any one of the previous claims, wherein the hops mixture includes hops pellets.

[0078] In some aspects, the techniques described herein relate to the method of any one of the previous claims, wherein the hops mixture includes hops cones.

[0079] In some aspects, the techniques described herein relate to the method of any one of the previous claims, wherein the hops mixture is contacted with ozone for up to 5 hours.

[0080] In some aspects, the techniques described herein relate to the method of any one of the previous claims, wherein the ozone is bubbled into the hops mixture.

[0081] In some aspects, the techniques described herein relate to the method of any one of the previous claims, wherein the ozone is contained in water, which is contacted with the hops mixture.

[0082] In some aspects, the techniques described herein relate to a composition including oxidized alpha acids prepared by the method of any one of the previous claims. In some aspects, the techniques described herein relate to a composition including greater than about 20 % by weight as determined by HPLC of total oxidized alpha acids.

[0083] In some aspects, the techniques described herein relate to a composition including greater than about 20 % by weight as determined by HPLC of total oxidized iso alpha acids.

[0084] In some aspects, the techniques described herein relate to a composition including greater than about 20 % by weight as determined by HPLC of total oxidized beta acids.

[0085] In some aspects, the techniques described herein relate to a composition including greater than about 20 % by weight as determined by HPLC of total oxidized hops compounds.

[0086] In some aspects, the techniques described herein relate to a composition, wherein the balance of the composition is water.

[0087] In some aspects, the techniques described herein relate to a composition, wherein the composition includes less than about 20 % as determined by HPLC of alpha and / or beta acids.

[0088] In some aspects, the techniques described herein relate to a supplement including oxidized alpha and beta acids.

[0089] In some aspects, the techniques described herein relate to a method of increasing fat burning, increasing weight loss, reducing weight gain, preventing fat accumulation, including: administering to a subject a composition including oxidized alpha and beta acids.

[0090] In some aspects, the techniques described herein relate to a method of inducing thennogenesis in brown adipose tissue, including: administering to a subject a composition including oxidized alpha and beta acids.

[0091] In some aspects, the techniques described herein relate to a method of supplementing an alcoholic beverage, including: adding oxidized alpha and beta acids to the alcoholic beverage.

Claims

CLAIMSWhat is claimed is:

1. A method of providing oxidized alpha and beta acids, comprising: contacting a hops mixture comprising alpha acids, beta acids, and / or iso-alpha acids with ozone, to thereby provide oxidized alpha and beta acids.

2. The method of claim 1, wherein the hops mixture is provided by contacting hops pellets with water under conditions to extract the alpha acids, beta acids, and / or iso-alpha acids from the hops pellets.

3. The method of any one of the previous claims, further comprising filtering the water.

4. The method of any one of the previous claims, wherein the hops mixture comprises hops pellets.

5. The method of any one of the previous claims, wherein the hops mixture comprises hops cones.

6. The method of any one of the previous claims, wherein the hops mixture is contacted with ozone for up to 5 hours.

7. The method of any one of the previous claims, wherein the ozone is bubbled into the hops mixture.

8. The method of any one of the previous claims, wherein the ozone is contained in water, which is contacted with the hops mixture.

9. A composition comprising oxidized alpha acids prepared by the method of any one of the previous claims.

10. A composition comprising greater than about 20 % by weight as determined by HPLC of total oxidized alpha acids.

11. A composition comprising greater than about 20 % by weight as determined by HPLC of total oxidized iso alpha acids.

12. A composition comprising greater than about 20 % by weight as determined by HPLC of total oxidized beta acids.

13. A composition comprising greater than about 20 % by weight as determined by HPLC of total oxidized hops compounds.

14. The composition of any one of claims 9-13, wherein the balance of the composition is water.

15. The composition of any one of claims 9-13, wherein the composition comprises less than about 20 % as determined by HPLC of alpha and / or beta acids.

16. A supplement comprising oxidized alpha and beta acids.

17. A method of increasing fat burning, increasing weight loss, reducing weight gain, preventing fat accumulation, comprising: administering to a subject a composition comprising oxidized alpha and beta acids.

18. A method of inducing thermogenesis in brown adipose tissue, comprising: administering to a subject a composition comprising oxidized alpha and beta acids.

19. A method of supplementing an alcoholic beverage, comprising: adding oxidized alpha and beta acids to the alcoholic beverage.

Images